Antibodies with fc modifications and methods of use thereof

Abstract

The present disclosure features anti-tumor necrosis factor receptor superfamily (TNFRSF), anti-tumor necrosis factor superfamily (TNFSF), anti-CD28, and anti-ICOS antibodies and antigen-binding fragments thereof having amino acid modifications at the Fc domain, and uses of these antibodies or antigen-binding fragments for modulating immune responses. The antibodies and antigen-binding fragments thereof can be used to treat various cancers, autoimmune disorders, neurological disorders, infectious diseases, inflammatory diseases, and transplant rejection.

Description

[0001] sequence list

[0002] This application contains a sequence list that has been submitted electronically in XML format and is hereby incorporated by full reference. The XML copy created on November 26, 2024, is named “00786-592WO5_Sequence_Listing_11_26_24” and has a size of 1,383,907 bytes. Background Technology

[0003] Maintaining control over cell-mediated and humoral immune responses is a crucial aspect of a healthy immune system. Aberrant regulation of lymphocyte-driven immune responses (e.g., T-cell and B-cell-driven immune responses) is associated with a wide range of human diseases, as inappropriate confinement of immune responses against various self- and foreign antigens plays a causal role in pathologies such as autoimmune diseases, infectious diseases, inflammatory diseases, neurological disorders or conditions, allergies, graft-versus-host disease, transplant rejection, and various other immune disorders. On the other hand, the use of immunotherapy is a prominent example of improving human pathologies involving inadequate or excessive immune responses. Targeting intracellular signaling pathways is a promising direction for the development of therapeutic agents for immunotherapy.

[0004] Proteins belonging to the tumor necrosis factor receptor superfamily (TNFRSF) and its ligands (TNFSF) are closely related to the activation, differentiation, and survival of immune system cells. TNFRSF members include, but are not limited to, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TNFR1, TNFR2, TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3. TNFSF members include, but are not limited to, TNF-α, TNF-β, lymphotoxin-β (LT-β), CD40L, FasL, CD30L, 4-1BBL, CD27L, OX40L, TRAIL, LIGHT, RANKL, TWEAK, APRIL, BAFF, VEGI, EDA-A1, EDA-A2, and GITRL. TNFRSF member proteins are oligomeric type I or III transmembrane proteins containing multiple extracellular cysteine-rich domains. Several of these receptors contain an intracellular death domain (DD), which, upon ligand binding, recruits caspase-interacting proteins to initiate caspase-activated extrinsic pathways. Other TNF superfamily receptors lacking death domains bind to TNF receptor-associated factor (TRAF) and activate intracellular signaling pathways (such as the NF-κB pathway), potentially leading to proliferation or differentiation in various cell types. In addition to these functions, several TNF superfamily receptors are also involved in regulating immune cell functions, such as B cell homeostasis and activation, natural killer cell activation, and T cell co-stimulation. Signaling of TNFRSF member proteins (such as 4-1BB and OX40) and other co-stimulatory proteins (such as CD28 and ICOS) is important in the regulation of immunity during inflammation and cancer.

[0005] Various antibodies or antigen-binding fragments with agonistic or antagonistic activity that specifically bind to TNFRSF and TNFSF member proteins, as well as CD28 and ICOS, have been described in the art. It is beneficial for therapeutic antibodies or antigen-binding fragments (e.g., antibodies or antigen-binding fragments binding to TNFRSF member proteins, TNFSF member proteins, CD28, or ICOS) to have a wide range of tolerable doses and dosing frequencies to allow for different dosing regimens to treat specific diseases or conditions in different patient populations. It is also important to ensure that antibodies or antigen-binding fragments (e.g., antibodies or antigen-binding fragments binding to TNFRSF member proteins, TNFSF member proteins, CD28, or ICOS) can be administered at high doses or dosing frequencies without loss of efficacy, as some studies have indicated that certain antibodies lose efficacy at high doses due to ineffective binding conformations (see, for example, Mayes et al., Nat. Rev. Drug Discov. 17:509-527, 2018; incorporated herein by reference). Therefore, there remains a great need for anti-TNFRSF antibodies, anti-TNFSF antibodies, anti-CD28 antibodies, or anti-ICOS antibodies, or their antigen-binding fragments, which can be administered at higher doses or frequencies and over a wide dose range to subjects (e.g., human subjects) and be tolerated by them, thereby demonstrating an improved treatment profile. Summary of the Invention

[0006] This disclosure is characterized by a method for modulating the immune response of a subject by administering a modified antibody or antigen-binding fragment thereof that specifically binds to a human TNFRSF member protein, a human TNFSF member protein, human CD28, or human ICOS. This disclosure also features anti-TNFRSF antibodies, anti-TNFSF antibodies, anti-CD28 antibodies, or anti-ICOS antibodies or antigen-binding fragments, and constructs thereof, wherein the antibody or antigen-binding fragment contains one or more mutations in its Fc domain, and the construct contains the antibody or antigen-binding fragment thereof. Furthermore, this disclosure features polynucleotides, vectors, and host cells that can be used to generate the antibody or antigen-binding fragment thereof or the construct, pharmaceutical compositions and kits containing the antibody or antigen-binding fragment thereof, constructs, polynucleotides, vectors, or host cells, and methods for using them to modulate the immune response of a subject.

[0007] In a first aspect, this disclosure is characterized by a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a human TNFRSF member protein, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof of this disclosure may be administered to a human subject over a wide dose range. The dose range over which the modified antibody or antigen-binding fragment thereof of this disclosure may be administered may be wider than, for example, the typical (or art-recognized) dose range of a corresponding unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof of this disclosure may be administered to a human subject over a dose range of about 10 mg to about 5000 mg or a dose range of about 1 mg / kg to about 50 mg / kg.

[0008] In a second aspect, this disclosure is characterized by a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a human TNFRSF member protein, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a more modulated dose and / or frequency that produces the immune response relative to an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency. In some embodiments, the modified antibody or antigen-binding fragment thereof may be administered to the human subject at a wide dose range (e.g., possibly a wider dose range than the typical (or art-recognized) dose range of the corresponding unmodified antibody or antigen-binding fragment thereof). The modified antibody or antigen-binding fragment thereof of this disclosure may be administered to the human subject once or more per month, every three weeks, every two weeks, weekly, every six days, every five days, every four days, every three days, every two days, or daily. The modified antibodies or antigen-binding fragments thereof disclosed herein may also be administered to human subjects at doses of about 10 mg to about 5000 mg or about 1 mg / kg to about 50 mg / kg. Compared to unmodified antibodies or antigen-binding fragments thereof, the modified antibodies or antigen-binding fragments thereof disclosed herein may produce greater (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) regulation (e.g., increased or enhanced; or decreased or suppressed) of immune responses (e.g., immune responses against cancer, such as cytotoxicity and cytokine production; immune responses against infection, such as antibody production, cytokine production and cytotoxicity; immune responses related to inflammation, such as cytokine production and eosinophil activation; or immune responses related to autoimmunity, etc.). When the modified antibodies or antigen-binding fragments of this disclosure are administered to human subjects at high doses and / or high dosing frequencies, the effect of the modified antibodies or antigen-binding fragments on the regulation of the immune response can be observed, wherein the high doses and / or high dosing frequencies are doses or dosing frequencies higher than the typical (or art-recognized) doses or dosing frequencies of the corresponding unmodified antibodies or antigen-binding fragments. Methods for assessing the regulation of the immune response are known in the art.See, for example, Clay et al., Clin. Cancer Res. 7:1127-1135, 2001; Van der Burg et al., Sci. Transl. Med. 3:108ps44, 2011; Albert-Vega et al., Front. Immunol. 9:2367, 2018; all incorporated herein by reference. In some embodiments, the modified antibody or its antigen-binding fragment is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or its antigen-binding fragment. A higher dose and / or frequency is defined as a dose or frequency higher than the typical (or art-recognized) dose or frequency of administration of the unmodified antibody or its antigen-binding fragment.

[0009] In some embodiments, the TNFRSF member proteins are selected from the group consisting of: TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3.

[0010] In some embodiments, the method further includes determining (e.g., measuring) the pre-treatment level of soluble TNFRSF member proteins in the serum or plasma of the human subject prior to administration of one or more doses of the modified antibody or its antigen-binding fragment thereof. In some embodiments, the method further includes determining (e.g., measuring) the post-treatment level of soluble TNFRSF member proteins in the serum or plasma of the human subject after administration of one or more doses of the modified antibody or its antigen-binding fragment thereof. In some embodiments, the method further includes determining (e.g., measuring) a reference level of soluble TNFRSF member proteins in the serum or plasma of a healthy person. In some cases, a reference level in the subject is determined before or after administration of the antibody or its antigen-binding fragment disclosed herein, the construct disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the host cell disclosed herein, or the pharmaceutical composition disclosed herein. For example, some non-limiting examples of reference levels for soluble TNFRSF member proteins include levels of soluble TNFRSF member proteins in subjects who have not been diagnosed with a disease, do not have symptoms of at least two or more diseases, or have not been given the anti-TNFRSF antibody or its antigen-binding fragment disclosed herein. An exemplary reference level for soluble TNFR2 (sTNFR2) in the serum of healthy individuals is approximately 2.51 ng / ml (see, for example, Kartikasari et al. Front. Immunol. 16:13:918254, 2022; incorporated herein by reference).

[0011] In some embodiments, the modified antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein, and:

[0012] (a) The pre-treatment level of the soluble TNFRSF member protein in the human subject was lower than or the same as the reference level of the soluble TNFRSF member protein; and / or

[0013] (b) The post-treatment level of the soluble TNFRSF member protein in the human subject is higher than the reference level or the pre-treatment level of the soluble TNFRSF member protein, and the method further comprises administering one or more additional doses of the agonist antibody or its antigen-binding fragment to the human subject during one or more additional treatment periods to maintain the higher level.

[0014] In some embodiments, the modified antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein, and:

[0015] (a) The human subject has a pre-treatment level of the soluble TNFRSF member protein that is higher than the reference level of the soluble TNFRSF member protein; and / or

[0016] (b) The post-treatment level of the soluble TNFRSF member protein in the human subject is lower than the reference level or the pre-treatment level of the soluble TNFRSF member protein, and the method further comprises administering one or more additional doses of the antagonistic antibody or its antigen-binding fragment to the human subject during one or more additional treatment periods to maintain the lower level.

[0017] In some embodiments, the modified antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

[0018] In some implementations, the TNFRSF member protein is TNFR2.

[0019] In some embodiments, the modified antibody or its antigen-binding fragment comprises a heavy chain complementarity-determining region (CDR) 1 (CDR-H1), wherein the CDR-H1 contains X 1 X 2 X 2 X 1 X 2 X 3 X 2 The amino acid sequence of JJJ, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X 2 Independently, it is Y, S, T, C, N, or Q; X 3 It is either D or E; and each J is independently a naturally occurring amino acid or is absent. In some embodiments, the CDR-H1 comprises GYTFTZ. 1 Z 2 The amino acid sequence of JJJ (SEQ ID NO: 1480), where Z 1 It is D or T; Z 2 It is Y, F, or L; and each J is independently a naturally occurring amino acid or is absent. In some embodiments, the CDR-H1 comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H2, CDR-H3, light chain CDR 1 (CDR-L1), CDR-L2, and CDR-L3 shown in Table 2.

[0020] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H1, wherein CDR-H1 comprises X 1 X 2 X 2 X 3 X 1 The amino acid sequence, where each X 1 Independently, it is either D or E; each X 2 Independently, it is Y, S, T, C, N, or Q; and X 3 It is L, A, V, G, I, M, W, F, or P. In some embodiments, the CDR-H1 comprises the amino acid sequence of DYNLD (SEQ ID NO: 1541). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0021] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H2, wherein CDR-H2 comprises X 1 X 2 X 3 X 2 X 3 X 3 X 1 X 3 X 3 X 3 X 3 X 3 X 3 X 4 X 2 X 4 X 2 The amino acid sequence, where each X 1 Independently, it is either D or E; each X 2 Independently, it is I, A, V, L, G, M, W, F, or P; each X 3 Independently, it is N, S, T, C, Y, or Q; and each X 4 Independently, it is K, R, or H. In some embodiments, the CDR-H2 comprises the amino acid sequence of DINPNYDSTSYSQKFRG (SEQ ID NO: 1542). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0022] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H3, wherein CDR-H3 comprises X 1X 2 X 2 X 1 X 2 X 1 X 3 X 1 The amino acid sequence, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X 2 Independently, it is N, S, T, C, Y, or Q; and X 3 It is D or E. In some embodiments, the CDR-H3 comprises the amino acid sequence of GNSWYFDV (SEQ ID NO: 1543). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2. In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-L1, which contains X 1 X 2 X 1 X 1 X 1 X 2 X 3 X 4 X 1 X 1 The amino acid sequence, where each X 1 Independently, it is S, Y, T, C, N, or Q; each X 2 Independently, it is A, G, V, L, I, M, W, F, or P; X 3 It is R, H, or K; and X 4 It is Y, A, V, I, L, M, F, or W. In some embodiments, the CDR-L1 comprises the amino acid sequence SASSSVRYNY (SEQ ID NO: 1544). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L2, and CDR-L3 shown in Table 2.

[0023] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-L2, wherein the CDR-L2 comprises X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is L, A, V, G, I, M, W, F, or P; and each X2 Independently, it is T, S, C, Y, N, or Q. In some embodiments, the CDR-L2 comprises the amino acid sequence of LTSNLAS (SEQ ID NO: 1545). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L3 shown in Table 2.

[0024] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-L3, wherein the CDR-L3 comprises X 1 X 2 X 2 X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is P, A, V, L, I, M, W, F, or G; and each X 2 Independently, it is Q, S, T, C, N, or Y. In some embodiments, the CDR-L3 comprises the amino acid sequence of PQQWSSNPLT (SEQ ID NO: 1546). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L2 shown in Table 2.

[0025] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as shown in Table 2.

[0026] In some embodiments, the modified antibody or its antigen-binding fragment comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH and the VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to the sequences shown in Table 3.

[0027] In some implementations, the TNFRSF member proteins are selected from the group consisting of CD27, CD40, GITR, OX40, and 4-1BB.

[0028] In some embodiments, the modified antibody or its antigen-binding fragment comprises the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of an antibody selected from the group consisting of: varlilumab (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Tavolimab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utomilumab (PF-05082566), and Urelumab (BMS-663513).

[0029] In some embodiments, the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and VL sequences of an antibody selected from the group consisting of: varirubin (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Talixicizumab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utoluumab (PF-05082566), and Ureluumab (BMS-663513).

[0030] In some embodiments, the serum or plasma level of the soluble TNFRSF member protein in the human subject is lower than the reference level of the soluble TNFRSF member protein.

[0031] In some embodiments, the modified antibody or its antigen-binding fragment exerts one or more biological activities selected from the group consisting of:

[0032] (a) Promotes NFκB activation;

[0033] (b) Inducing and regulating the proliferation of T (T-reg) cell populations;

[0034] (c) Directly kills a population of CD8+ T cells or promotes the death of said population;

[0035] (d) Promote the increase in the level of one or more mRNA molecules encoding proteins selected from the group consisting of the following (e.g., an increase of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more): cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in the angiogenesis pathway, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α and lymphotoxin β;

[0036] (e) Promotes an increase in the level of one or more proteins selected from the group consisting of the following (e.g., an increase of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more): cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in the angiogenesis pathway, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α and lymphotoxin β;

[0037] (f) Promotes an increase in itaconic acid levels (e.g., an increase of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more); and

[0038] (g) Promotes an increase in the level of the soluble TNFRSF member protein (e.g., an increase of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more).

[0039] In some embodiments, the modified antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

[0040] In some implementations, the TNFRSF member protein is TNFR2.

[0041] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as shown in Table 4.

[0042] In some embodiments, the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the sequences shown in Table 5.

[0043] In some embodiments, the serum or plasma level of the soluble TNFRSF member protein in the human subject is higher than the reference level of the soluble TNFRSF member protein.

[0044] In some embodiments, the modified antibody or its antigen-binding fragment exerts one or more biological activities selected from the group consisting of:

[0045] (a) Inhibit NFκB activation;

[0046] (b) Reduce or inhibit the proliferation of T-reg cell populations;

[0047] (c) Inducing the proliferation of CD8+ T cell populations;

[0048] (d) Reduce or inhibit the proliferation of cancer cell populations;

[0049] (e) Inhibits TNFRSF member protein signaling in proliferating cells;

[0050] (f) It does not inhibit TNFRSF member protein signaling in resting cells;

[0051] (g) Reduce or inhibit the proliferation of myeloid-derived suppressor cells (MDSCs);

[0052] (h) Promotes a reduction in the level of one or more mRNA molecules encoding proteins selected from the group consisting of the following (e.g., a reduction of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more): cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in the angiogenesis pathway, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α and lymphotoxin β;

[0053] (i) Promotes a reduction in the level of one or more proteins selected from the group consisting of (e.g., a reduction of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more): cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in angiogenesis pathways, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α and lymphotoxin β; and

[0054] (j) Promotes a reduction in the level of the soluble TNFRSF member protein (e.g., a reduction of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more).

[0055] In some embodiments, the modified antibody or its antigen-binding fragment is neither an agonist nor an antagonist of the TNFRSF member protein.

[0056] In some implementations, the TNFRSF member protein is TNFR2.

[0057] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as shown in Table 6.

[0058] In some embodiments, the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the sequences shown in Table 7.

[0059] In some embodiments, the modified antibody or its antigen-binding fragment has less than about 90% (e.g., less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10%) cross-reactivity with the soluble TNFRSF member protein. In some embodiments, the modified antibody or its antigen-binding fragment has less than 50% cross-reactivity with the soluble TNFRSF member protein. For example, the antibody or its antigen-binding fragment may bind to TNFR2 but exhibit less than 50% (e.g., less than 40%, 30%, 20%, or 10%) cross-reactivity with soluble TNFR2. The cross-reactivity of the antibody or its antigen-binding fragment with the antigen can be determined using methods known in the art, such as by assessing the percentage of homology between the antigen sequence of the antibody-bound protein and the sequence of a second protein. A high percentage of homology indicates that the antibody may cross-react with the second protein. The cross-reactivity of antibodies or their antigen-binding fragments with antigens can be determined using biophysical techniques such as surface plasmon resonance (SPR) assays or Biacore binding assays.

[0060] In a third aspect, this disclosure is characterized by a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a human TNFSF member protein, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof may be administered to the human subject at a wide dose range (e.g., possibly a wider dose range than the typical (or art-recognized) dose range of the corresponding unmodified antibody or antigen-binding fragment thereof).

[0061] In a fourth aspect, this disclosure features a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a human TNFSF member protein, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a greater modulated dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency to produce the immune response. In some embodiments, the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof may be administered to the human subject at a wide dose range (e.g., possibly a wider dose range than the typical (or art-recognized) dose range of the corresponding unmodified antibody or antigen-binding fragment thereof).

[0062] In some implementations, the TNFSF member protein is TRAIL.

[0063] In a fifth aspect, this disclosure is characterized by a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human CD28, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof may be administered to the human subject at a wide dose range (e.g., possibly a wider dose range than the typical (or art-recognized) dose range of the corresponding unmodified antibody or antigen-binding fragment thereof).

[0064] In a sixth aspect, this disclosure features a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human CD28, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a greater modulated dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency to produce the immune response. In some embodiments, the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof may be administered to the human subject at a wide dose range (e.g., possibly a wider dose range than the typical (or art-recognized) dose range of the corresponding unmodified antibody or antigen-binding fragment thereof).

[0065] In some embodiments, the modified antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to CD28. In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of theralizumab (TAB-08). In some embodiments, the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical to the VH and VL sequences of theralizumab (TAB-08).

[0066] In a seventh aspect, this disclosure is characterized by a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human ICOS, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof may be administered to the human subject at a wide dose range (e.g., possibly a wider dose range than the typical (or art-recognized) dose range of the corresponding unmodified antibody or antigen-binding fragment thereof).

[0067] In an eighth aspect, this disclosure features a method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human ICOS, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and further wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a greater modulated dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency that produces the immune response. In some embodiments, the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof. In some embodiments, the modified antibody or antigen-binding fragment thereof may be administered to the human subject at a wide dose range (e.g., possibly a wider dose range than the typical (or art-recognized) dose range of the corresponding unmodified antibody or antigen-binding fragment thereof).

[0068] In some embodiments, the modified antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to ICOS. In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of GSK-3359609 or JTX-2011. In some embodiments, the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical to the VH and VL sequences of GSK-3359609 or JTX-2011.

[0069] In some embodiments, one or more doses of the modified antibody or its antigen-binding fragment are administered to the human subject during one or more treatment periods, wherein each dose comprises about 0.1 mg to about 5000 mg of the modified antibody or its antigen-binding fragment. In some embodiments, each dose comprises about 0.001 mg / kg to about 50 mg / kg of the modified antibody or its antigen-binding fragment. In some embodiments, each treatment period lasts for one week, two weeks, three weeks, four weeks, five weeks, six weeks, seven weeks, eight weeks, or longer.

[0070] In some implementations, the modified antibody or its antigen-binding fragment is administered to the human subject once or more per month, every three weeks, every two weeks, weekly, every six days, every five days, every four days, every three days, every two days, or daily.

[0071] In some embodiments, the modified antibody or its antigen-binding fragment is administered to the human subject at a dose of about 10 mg to about 5000 mg. In some embodiments, the modified antibody or its antigen-binding fragment is administered to the human subject at a dose of about 1 mg / kg to about 50 mg / kg.

[0072] In some embodiments, the one or more amino acid modifications comprise one or more substitutions, deletions, insertions, or chemical modifications of the Fc domain.

[0073] In some embodiments, the modified antibody or its antigen-binding fragment is selected from the group consisting of: monoclonal antibodies or their antigen-binding fragments, polyclonal antibodies or their antigen-binding fragments, human antibodies or their antigen-binding fragments, humanized antibodies or their antigen-binding fragments, primate-derived antibodies or their antigen-binding fragments, bispecific antibodies or their antigen-binding fragments, multispecific antibodies or their antigen-binding fragments, bivariate immunoglobulin domains, monovalent antibodies or their antigen-binding fragments, and chimeric antibodies or their antigen-binding fragments. In some embodiments, the modified antibody or its antigen-binding fragment is a human antibody, a humanized antibody, or a chimeric antibody, or their antigen-binding fragments.

[0074] In some embodiments, the modification of one or more amino acids at the Fc domain reduces (e.g., by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or more) the binding affinity of the modified antibody or antigen-binding fragment thereof to the Fc receptor relative to the corresponding unmodified antibody or antigen-binding fragment thereof. The reduction in binding affinity of the antibody or antigen-binding fragment thereof to the Fc receptor of this disclosure (e.g., by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or more) can manifest as an equilibrium dissociation constant (K0) between the antibody or antigen-binding fragment thereof and its antigen (e.g., a TNFRSF member protein). D The K ratio between the unmodified antibody or its antigen-binding fragment and the antigen is... DHigh (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more). The K-value between the antibody or its antigen-binding fragment and the antigen. D The effector function can be measured using methods such as surface plasmon resonance (SPR) (e.g., using a Biacore™ system). In some embodiments, the effector function of the modified antibody or its antigen-binding fragment is independent of antibody-dependent cytotoxicity (ADCC) or antibody-dependent phagocytosis (ADCP). The effector function (e.g., ADCC or ADCP) of the antibody or its antigen-binding fragment disclosed herein can be determined using methods known in the art (see, for example, Parekh et al. MAbs 4:310-318, 2012; and Kamen et al. J. Immunol. Methods 468:55-60, 2019). In some embodiments, the modification of one or more amino acids at the Fc domain enhances the biological activity of the modified antibody or its antigen-binding fragment relative to the corresponding unmodified antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment exhibits dimer binding to its antigen (e.g., TNFRSF member protein, TNFSF member protein, CD28, or ICOS) at high doses, wherein the modified antibody or its antigen-binding fragment comprises a first arm and a second arm, further wherein the first arm binds to the antigen, and the second arm binds to an adjacent protein (e.g., an adjacent antigen of the antibody or its antigen-binding fragment). In some embodiments, the modified antibody or its antigen-binding fragment forms a hexagonal network with its antigen (e.g., TNFRSF member protein, TNFSF member protein, CD28, or ICOS) at high doses.

[0075] In some embodiments, the Fc receptor is selected from the group consisting of FcγRI, FcγRII, and FcγRIII. In some embodiments, the biological activity of the modified antibody or its antigen-binding fragment is independent of ADCC or ADCP. In some embodiments, the adjacent protein is an antigen of the antibody or its antigen-binding fragment (e.g., a TNFRSF member protein, a TNFSF member protein, CD28, or ICOS).

[0076] In some implementations, the Fc domain is selected from the group consisting of: IgG Fc domain, IgAFc domain, IgD Fc domain, IgE Fc domain and IgM Fc domain.

[0077] In some implementations, the Fc domain is an IgG Fc domain.

[0078] In some embodiments, the Fc domain is the human IgG1 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0079] (i) L235E;

[0080] (ii) L234A and L235A;

[0081] (iii) L234A, L235A and P329G;

[0082] (iv) L234E, L235F and P331S;

[0083] (v)D265A;

[0084] (vi) G237A;

[0085] (vii) E318A;

[0086] (viii) E233P;

[0087] (ix) G236R and L328R;

[0088] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0089] (xi) N297G or N297Q,

[0090] The amino acid positions are based on the EU index number.

[0091] In some embodiments, the modified antibody or its antigen-binding fragment is an IgG1 antibody or its antigen-binding fragment.

[0092] In some embodiments, the Fc domain is the human IgG2 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0093] (i) D265A;

[0094] (ii) G237A;

[0095] (iii) E318A;

[0096] (iv) H268Q, V309L, A330S and P331S;

[0097] (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and

[0098] (vi) N297G or N297Q,

[0099] The amino acid positions are based on the EU index number.

[0100] In some embodiments, the modified antibody or its antigen-binding fragment is an IgG2 antibody or its antigen-binding fragment.

[0101] In some embodiments, the modified antibody or its antigen-binding fragment is an IgG3 antibody or its antigen-binding fragment.

[0102] In some embodiments, the Fc domain is the human IgG3 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0103] (i) L235E;

[0104] (ii) L234A and L235A;

[0105] (iii) L234A, L235A and P329G;

[0106] (iv) L234E, L235F and P331S;

[0107] (v)D265A;

[0108] (vi) G237A;

[0109] (vii) E318A;

[0110] (viii) E233P;

[0111] (ix) G236R and L328R;

[0112] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0113] (xi) N297G or N297Q,

[0114] The amino acid positions are based on the EU index number.

[0115] In some embodiments, the Fc domain is the human IgG4 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0116] (i) L235E;

[0117] (ii) D265A;

[0118] (iii) G237A;

[0119] (iv) E318A;

[0120] (v)E233P;

[0121] (vi) G236R and L328R; and

[0122] (vii) N297G or N297Q,

[0123] The amino acid positions are based on the EU index number.

[0124] In some embodiments, the modified antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P (according to Kabat's S241P). In some embodiments, the one or more amino acid modifications are one or more amino acid substitutions selected from the group consisting of:

[0125] (i) L235E; and

[0126] (ii) F234A and L235A,

[0127] The amino acid positions are based on the EU index number.

[0128] In some embodiments, the modified antibody or its antigen-binding fragment is an IgG4 antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment contains an amino acid substitution for S228P.

[0129] In some embodiments, the modified antibody or its antigen-binding fragment does not contain any of the amino acid substitutions shown in Table 18, wherein the amino acid positions are based on EU index numbers.

[0130] In some embodiments, the one or more amino acid modifications are the deletion of one or more amino acids. In some embodiments, the one or more amino acid deletions are the deletion of all amino acid residues of the Fc domain. In some embodiments, the modified antibody or its antigen-binding fragment lacks the Fc domain. In some embodiments, the modified antibody or its antigen-binding fragment is selected from the group consisting of: single-chain Fv molecules (scFv), bifunctional antibodies, trifunctional antibodies, antibody-like protein scaffolds, Fv fragments, Fab fragments, F(ab')2 molecules, and tandem scFv (taFv).

[0131] In some embodiments, the modified antibody or its antigen-binding fragment is conjugated to a therapeutic agent. In some embodiments, the therapeutic agent is selected from the group consisting of: chemotherapeutic agents, immunotherapeutic agents, agonists of TNFRSF member proteins, antagonists of TNFRSF member proteins, agonists of TNFSF member proteins, antagonists of TNFSF member proteins, agonists of CD28, antagonists of CD28, agonists of ICOS, and antagonists of ICOS.

[0132] In some embodiments, the method includes administering an additional therapeutic agent. In some embodiments, the additional therapeutic agent is selected from the group consisting of: chemotherapeutic agents, immunotherapeutic agents, agonists of TNFRSF member proteins, antagonists of TNFRSF member proteins, agonists of TNFSF member proteins, antagonists of TNFSF member proteins, agonists of CD28, antagonists of CD28, agonists of ICOS, and antagonists of ICOS.

[0133] In some embodiments, the modified antibody or its antigen-binding fragment is administered intravenously or subcutaneously to the human subject. In some embodiments, the modified antibody or its antigen-binding fragment is administered subcutaneously to the human subject.

[0134] In some implementations, the method suppresses the immune response mediated by B cells, CD8+ T cells, or T-reg cells in the human subject.

[0135] In some implementations, the method promotes an immune response in the human subject mediated by B cells, CD8+ T cells, or T-reg cells.

[0136] In some embodiments, the human subject requires tissue or organ regeneration. In some embodiments, the tissue or organ is selected from the group consisting of: pancreas, islets of Langerhans, salivary glands, pituitary gland, kidney, heart, lung, hematopoietic system, cranial nerves, heart, aorta, olfactory glands, ear, nerves, head structures, eye, thymus, tongue, bone, liver, small intestine, large intestine, intestine, lung, brain, skin, peripheral nervous system, central nervous system, spinal cord, mammary glands, embryonic structures, embryo, and testis.

[0137] In some embodiments, the method is used to treat a cell proliferation disorder in the human subject. In some embodiments, the cell proliferation disorder is a cancer selected from the group consisting of: leukemia, lymphoma, liver cancer, bone cancer, lung cancer, brain cancer, bladder cancer, gastrointestinal cancer, breast cancer, heart cancer, cervical cancer, uterine cancer, head and neck cancer, gallbladder cancer, laryngeal cancer, lip and oral cavity cancer, eye cancer, melanoma, pancreatic cancer, prostate cancer, colorectal cancer, testicular cancer, and pharyngeal cancer. In some implementations, the cell proliferation disorder is a cancer selected from the group consisting of: acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), adrenocortical carcinoma, AIDS-related lymphoma, primary central nervous system (CNS) lymphoma, anal cancer, appendiceal cancer, astrocytoma, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, extrahepatic cancer, Ewing sarcoma family, osteosarcoma and malignant fibrous histiocytoma, embryonal tumors of the central nervous system, germ cell tumors of the central nervous system, craniopharyngioma, ependymoma, bronchial tumor, Burkitt's lymphoma. Lymphoma, carcinoid tumor, primary lymphoma, chordoma, chronic myeloproliferative neoplasm, colon cancer, extrahepatic bile duct carcinoma, ductal carcinoma in situ (DCIS), endometrial cancer, ependymoma, esophageal cancer, olfactory neuroblastoma, extracranial germ cell tumor, gonadal germ cell tumor, fallopian tube cancer, osteofibrous histiocytoma, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), testicular germ cell tumor, gestational trophoblastic disease, glioma, childhood brainstem glioma, hairy cell leukemia, hepatocellular carcinoma, Langerhans cell histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, islet cell tumor, pancreatic neuroendocrine tumor, Wilms tumor and other childhood renal tumors, Langerhans cell histiocytosis, small cell lung cancer, cutaneous T-cell lymphoma, intraocular melanoma, Merkel cell carcinoma. Carcinoma, mesothelioma, metastatic squamous neck carcinoma, midline carcinoma, multiple endocrine adenoma syndrome, multiple myeloma / plasma cell tumor (e.g., multiple myeloma or refractory multiple myeloma), myelodysplastic syndrome, nasal cavity and sinus carcinoma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin lymphoma (non-Hodgkin lymphoma),NHL), non-small cell lung cancer (NSCLC), epithelial ovarian cancer, germ cell ovarian cancer, low-grade malignant potential ovarian cancer, pancreatic neuroendocrine tumors, papilloma, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid carcinoma, penile cancer, pharyngeal cancer, pheochromocytoma, pituitary adenoma, pleural pulmonary blastoma, primary peritoneal cancer, rectal cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, Kaposi sarcoma, rhabdomyosarcoma, Sézary syndrome, small bowel cancer, soft tissue sarcoma, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, urethral cancer, endometrial uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, and Waldenström macroglobulinemia. In some embodiments, the modified antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to a TNFRSF member protein selected from the group consisting of: TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TNFR1, Fas, CD40, CD27, 4-1BB, OX40, GITR, and XEDAR. In some embodiments, the modified antibody or its antigen-binding fragment is an antagonist antibody or its antigen-binding fragment that specifically binds to a TNFRSF member protein selected from the group consisting of: TRAMP, NGFR, TRAIL-R4, TNFR2, HVEM, CD30, TROY, and RELT. In some embodiments, the modified antibody or its antigen-binding fragment is an antagonist antibody or its antigen-binding fragment that specifically binds to TNFR2 comprising: CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 as shown in Table 4. In some embodiments, the modified antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to TNFR2 comprising VH and VL, wherein VH and VL have an amino acid sequence that is at least 80% identical to the sequences shown in Table 5.

[0138] In some implementations, the method is used to treat autoimmune diseases or inflammatory diseases in the human subject. In some implementations, the autoimmune disease is selected from the group consisting of: type 1 diabetes mellitus, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, Behçet's disease, bullous pemphigoid, cardiomyopathy, celiac dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, localized scleroderma (CREST syndrome), cold agglutinin disease, Crohn's disease, discoid lupus erythematosus, primary mixed cryoglobulinemia vasculitis, fibromyalgia-fibromyositis, Graves' disease, and Guillain-Barré syndrome. Hashimoto's thyroiditis, hypothyroidism, inflammatory bowel disease, autoimmune lymphoproliferative syndrome (ALPS), idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, juvenile arthritis, lichen planus, lupus, lupus nephritis, Ménière's disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychondritis, polygonococcal syndrome, polymyalgia rheumatica, polymyositis, dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome This includes conditions such as stiff-man syndrome, Takayasu's arteritis, temporal arteritis / giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, Wegener's granulomatosis, Goodpasture syndrome, transplant rejection, celiac disease, esophagitis, and inflammatory central nervous system disorders. In some embodiments, the inflammatory central nervous system disorder is Alzheimer's disease.

[0139] In some embodiments, the method is used to treat an infectious disease in the human subject. In some embodiments, the infectious disease is caused by one or more infectious agents selected from the group consisting of viruses, bacteria, fungi, and parasites. In some implementations, the infectious disease is caused by a virus selected from the group consisting of: hepatitis C virus, yellow fever virus, Kadam virus, Kyasanur Forest disease virus, Langat virus, Omsk hemorrhagic fever virus, Powassan virus, Royal Farm virus, Karshi virus, tick-borne encephalitis virus, Neudoerfl virus, Sofjin virus, sheep jumping disease virus, Negishi virus, Meaban virus, Saumarez Reef virus, Tyuleniy virus, Aroa virus, dengue virus, Kedougou virus, Cacipacore virus, Koutango virus, Japanese encephalitis virus. encephalitis virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Usutu virus, West Nile virus, Yaounde virus, Kokobera virus, Bagaza virus, Ilheus virus, Israel turkey meningoencephalitis-myelitis virus, Ntaya virus, Tembusu virus, Zika virus, Banzivirus, Bouboui virus, Edge Hill virus, Jugra virus, Saboya virus, Sepik virusviruses, Uganda S virus, Wesselsbron virus, yellow fever virus, Entebbebat virus, Yokose virus, Apoi virus, CowboneRidge virus, Jutiapa virus, Modoc virus, Sal Vieja virus, San Perlita virus, Bukalasabat virus, Carey Island virus, Dakar bat virus, Montana myotis leukoencephalitis virus, Phnom Penh bat virus, Rio Bravo virus, Tamana bat virus, cell fusion agent virus, Ippy virus, Lassa virus Lymphocytic choroidal meningitis virus (LCMV), Mobala virus, Mopeia virus, Amapari virus, Flexal virus, Guanarito virus, Junin virus, Latino virus, Machupo virus, Oliveros virus, Paraná virus, Pichinde virus, Pirital virus, Sabiá virus, Tacaribe virus, Tamiami virus, Whitewater Arroyo virus, Chapare virus, Lujo virus, Hantaanvirus, Sin Nombre virus, Dugbe virus, Bunyamwera virus virus, Rift Valley feverFever virus, La Crossevirus, California encephalitis virus, Crimean-Congo hemorrhagic fever (CCHF) virus, Ebola virus, Marburg virus, Venezuelan equine encephalitis virus (VEE), Eastern equine encephalitis virus (EEE), Western equine encephalitis virus (WEE), Sindbis virus, rubella virus, Semliki Forest virus, Ross River virus, Barmah Forest virus, O'nyong'nyong virus, chikungunya virus Smallpox virus, monkeypox virus, vaccinia virus, herpes simplex virus, human herpesvirus, cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella-zoster virus, Kaposi's sarcoma-associated herpesvirus (KSHV), influenza virus, severe acute respiratory syndrome (SARS) virus, rabies virus, vesicular stomatitis virus (VSV), human respiratory syncytial virus (RSV), Newcastle disease virus, hendra virus, nipah virus, measles virus, rinderpest virus, canine distemper virus, Sendai virus. Human parainfluenza virus (e.g., types 1, 2, 3, and 4), rhinovirus, mumps virus, poliovirus, human enterovirus (e.g., types A, B, C, and D), hepatitis A virus, coxsackievirus, hepatitis B virus, human papillomavirus, adeno-associated virus, astrovirus, JC virus, BK virus, SV40 virus, Norwalk virus.Virus, rotavirus, human immunodeficiency virus (HIV), and human T-lymphotropic virus types I and II. In some embodiments, the infectious disease is caused by bacteria belonging to genera selected from the group consisting of: Salmonella, Streptococcus, Bacillus, Listeria, Corynebacterium, Nocardia, Neisseria, Acinetobacter, Moraxella, Enterobacteriaceae, Pseudomonas, Escherichia, Klebsiella, Serratia, etc. The genera *Serratia*, *Enterobacter*, *Proteus*, *Salmonella*, *Shigella*, *Yersinia*, *Haemophilus*, *Bordetella*, *Legionella*, *Pasteurella*, *Francisella*, *Brucella*, *Bartonella*, *Clostridium*, *Vibrio*, *Campylobacter*, and *Staphylococcus*. In some embodiments, the infectious disease is caused by fungi belonging to genera selected from the group consisting of: *Aspergillus*, *Candida*, *Malassezia*, *Trichosporon*, *Fusarium*, *Acremonium*, *Rhizopus*, *Mucor*, *Pneumocystis*, and *Absidia*. In some embodiments, the infectious disease is caused by parasites selected from the group consisting of: *Entamoeba histolytica*, *Giardia lamblia*, *Cryptosporidium muris*, *Trypanosomatida gambiense*, and *Trypanosomatida rhodesiana*.Leishmania rhodesiense, Trypanosomatida crusi, Leishmania mexicana, Leishmania braziliensis, Leishmania tropica, Leishmania donovani, Toxoplasma gondii, Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, Plasmodium falciparum, Trichomonas vaginalis, and Histomonas meleagridis. Exemplary worm parasites include *Trichuristrichiura*, *Ascaris lumbricoides*, *Enterobius vermicularis*, *Ancylostoma duodenale*, *Necatora Americanus*, *Strongyloides stercoralis*, *Wuchereria bancrofti*, *Dracunculus medinensis*, *Schistosoma mansoni*, *Schistosoma haematobium*, *Schistosoma japonicum*, *Fasciola hepatica*, *Fasciola gigantica*, *Heterophyes*, and *Paragonimus*. Westermani, Taenia solium, Taenia saginata, Hymenolepis nana, and Echinococcus granulosus.

[0140] In some embodiments, the method is used to treat an inflammatory disease in the human subject. In some embodiments, the inflammatory disease is selected from the group consisting of: acute or chronic inflammation, cardiac fibrosis, pulmonary fibrosis, osteoarthritis, rheumatoid arthritis, atherosclerosis, type I diabetes, type II diabetes, graft-versus-host disease, multiple sclerosis, osteomyelitis, psoriasis, Crohn's disease, Sjögren's syndrome, lupus erythematosus, and ulcerative colitis.

[0141] In some embodiments, the method is used to treat a neurological disease or condition in the human subject. In some embodiments, the neurological disease or condition is selected from the group consisting of: brain tumors, brain metastases, brain injury, spinal cord injury, nerve injury, schizophrenia, epilepsy, Parkinson's disease, autism, Huntington's disease, stroke, Alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), and myasthenia gravis.

[0142] In some embodiments, the method is used to treat allergies in the human subject. In some embodiments, the allergy is selected from the group consisting of: food allergies, seasonal allergies, pet allergies, urticaria, hay fever, allergic conjunctivitis, poison ivy allergy, oak allergy, mold allergy, drug allergy, dust allergy, cosmetic allergy, and chemical allergy.

[0143] In some embodiments, the method is used to treat transplant rejection in the human subject. In some embodiments, the transplant rejection is allogeneic graft rejection. In some embodiments, the transplant rejection is selected from the group consisting of: skin graft rejection, bone graft rejection, vascular tissue graft rejection, ligament graft rejection, and organ graft rejection. In some embodiments, the ligament graft rejection is selected from the group consisting of: cricothyroid ligament graft rejection, periodontal ligament graft rejection, lens suspensory ligament graft rejection, radial carpal ligament graft rejection, radial carpal dorsal ligament graft rejection, ulnar collateral ligament graft rejection, radial collateral ligament graft rejection, breast suspensory ligament graft rejection, anterior sacroiliac ligament graft rejection, posterior sacroiliac ligament graft rejection, sacrotuberous ligament graft rejection, sacrospinous ligament graft rejection, infrapubic ligament graft rejection, suprapubic ligament graft rejection, anterior cruciate ligament graft rejection, lateral collateral ligament graft rejection, posterior cruciate ligament graft rejection, medial collateral ligament graft rejection, craniocarpal ligament graft rejection, caudal cruciate ligament graft rejection, and patellar ligament graft rejection. In some embodiments, the organ graft rejection is selected from the group consisting of: heart graft rejection, lung graft rejection, kidney graft rejection, liver graft rejection, pancreas graft rejection, intestinal graft rejection, and thymus graft rejection.

[0144] In some embodiments, the method is used to treat graft-versus-host disease in the human subject. In some embodiments, the graft-versus-host disease is caused by bone marrow transplantation or by one or more blood cells selected from the group consisting of: hematopoietic stem cells, common myeloid progenitor cells, common lymphoid progenitor cells, megakaryocytes, monocytes, basophils, eosinophils, neutrophils, macrophages, T cells, B cells, natural killer cells, and dendritic cells.

[0145] In some embodiments, the method is used for transplantation or for regeneration of tissues or organs of the human subject. In some embodiments, the tissues or organs are selected from the group consisting of: pancreas, islets of Langerhans, salivary glands, pituitary gland, kidneys, heart, lungs, hematopoietic system, cranial nerves, heart, aorta, olfactory glands, ear, nerves, eye, thymus, tongue, bone, liver, small intestine, large intestine, gastrointestinal tract, lungs, brain, skin, peripheral nervous system, central nervous system, spinal cord, mammary glands, embryonic structures, embryo, and testes. In some embodiments, the tissues or organs express the TNFRSF member protein, the TNFSF member protein, CD28, or ICOS.

[0146] In a ninth aspect, this disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to a human TNFRSF member protein, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, further wherein:

[0147] (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0148] (i) L235E;

[0149] (ii) L234A and L235A;

[0150] (iii) L234A, L235A and P329G;

[0151] (iv) L234E, L235F and P331S;

[0152] (v)D265A;

[0153] (vi) G237A;

[0154] (vii) E318A;

[0155] (viii) E233P;

[0156] (ix) G236R and L328R;

[0157] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0158] (xi) N297G or N297Q;

[0159] (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0160] (i) D265A;

[0161] (ii) G237A;

[0162] (iii) E318A;

[0163] (iv) H268Q, V309L, A330S and P331S;

[0164] (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and

[0165] (vi) N297G or N297Q;

[0166] (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0167] (i) L235E;

[0168] (ii) L234A and L235A;

[0169] (iii) L234A, L235A and P329G;

[0170] (iv) L234E, L235F and P331S;

[0171] (v)D265A;

[0172] (vi) G237A;

[0173] (vii) E318A;

[0174] (viii) E233P;

[0175] (ix) G236R and L328R;

[0176] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0177] (xi) N297G or N297Q;

[0178] (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0179] (i) L235E;

[0180] (ii) D265A;

[0181] (iii) G237A;

[0182] (iv) E318A;

[0183] (v)E233P;

[0184] (vi) G236R and L328R; and

[0185] (vii) N297G or N297Q; or

[0186] (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P (according to Kabat's S241P), the Fc domain being a human IgG4 Fc domain, and optionally the one or more amino acid substitutions being selected from the group consisting of:

[0187] (i) L235E; and

[0188] (ii) F234A and L235A,

[0189] Furthermore, the amino acid positions are based on EU index numbers.

[0190] In some embodiments, the antibody or its antigen-binding fragment is an IgG4 antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment contains an amino acid substitution for S228P.

[0191] In some embodiments, the TNFRSF member protein is selected from the group consisting of: TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3. In some embodiments, the TNFRSF member protein is TNFR2.

[0192] In some embodiments, the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

[0193] In some embodiments, the TNFRSF member protein is TNFR2. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, wherein CDR-H1 comprises X 1 X 2 X 2 X 1 X 2 X 3 X 2 The amino acid sequence of JJJ, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X 2 Independently, it is Y, S, T, C, N, or Q; X 3It is either D or E; and each J is independently a naturally occurring amino acid or is absent. In some embodiments, the CDR-H1 comprises GYTFTZ. 1 Z 2 The amino acid sequence of JJJ (SEQ ID NO: 1480), where Z 1 It is D or T; Z 2 It is Y, F, or L; and each J is independently a naturally occurring amino acid or is absent. In some embodiments, the CDR-H1 comprises the amino acid sequence GYTFTDY (SEQ ID NO: 1413). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0194] In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, wherein CDR-H1 comprises X 1 X 2 X 2 X 3 X 1 The amino acid sequence, where each X 1 Independently, it is either D or E; each X 2 Independently, it is Y, S, T, C, N, or Q; and X 3 It is L, A, V, G, I, M, W, F, or P. In some embodiments, the CDR-H1 comprises the amino acid sequence of DYNLD (SEQ ID NO: 1541). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0195] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H2, wherein CDR-H2 comprises X 1 X 2 X 3 X 2 X 3 X 3 X 1 X 3 X 3 X 3 X 3 X 3 X 3 X 4 X 2 X 4 X 2 The amino acid sequence, where each X 1Independently, it is either D or E; each X 2 Independently, it is I, A, V, L, G, M, W, F, or P; each X 3 Independently, it is N, S, T, C, Y, or Q; and each X 4 Independently, it is K, R, or H. In some embodiments, the CDR-H2 comprises the amino acid sequence of DINPNYDSTSYSQKFRG (SEQ ID NO: 1542). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0196] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-H3, wherein CDR-H3 comprises X 1 X 2 X 2 X 1 X 2 X 1 X 3 X 1 The amino acid sequence, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X 2 Independently, it is N, S, T, C, Y, or Q; and X 3 It is D or E. In some embodiments, the CDR-H3 comprises the amino acid sequence of GNSWYFDV (SEQ ID NO: 1543). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-L1, CDR-L2 and CDR-L3 shown in Table 2.

[0197] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-L1, wherein CDR-L1 comprises X 1 X 2 X 1 X 1 X 1 X 2 X 3 X 4 X 1 X 1 The amino acid sequence, where each X 1 Independently, it is S, Y, T, C, N, or Q; each X 2 Independently, it is A, G, V, L, I, M, W, F, or P; X 3 It is R, H, or K; and X 4It is Y, A, V, I, L, M, F, or W. In some embodiments, the CDR-L1 comprises the amino acid sequence SASSSVRYNY (SEQ ID NO: 1544). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L2, and CDR-L3 shown in Table 2.

[0198] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-L2, wherein the CDR-L2 comprises X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is L, A, V, G, I, M, W, F, or P; and each X 2 Independently, it is T, S, C, Y, N, or Q. In some embodiments, the CDR-L2 comprises the amino acid sequence of LTSNLAS (SEQ ID NO: 1545). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L3 shown in Table 2.

[0199] In some embodiments, the modified antibody or its antigen-binding fragment comprises CDR-L3, wherein the CDR-L3 comprises X 1 X 2 X 2 X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is P, A, V, L, I, M, W, F, or G; and each X 2 Independently, it is Q, S, T, C, N, or Y. In some embodiments, the CDR-L3 comprises the amino acid sequence of PQQWSSNPLT (SEQ ID NO: 1546). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L2 shown in Table 2.

[0200] In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 as shown in Table 2. In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the sequences shown in Table 3.

[0201] In some embodiments, the TNFRSF member proteins are selected from the group consisting of: CD27, CD40, GITR, OX40, and 4-1BB. In some embodiments, the antibody or its antigen-binding fragment comprises the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of an antibody selected from the group consisting of: varirubin (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Talixicizumab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utoluumab (PF-05082566), and Ureluumab (BMS-663513). In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical to the VH and VL sequences of an antibody selected from the group consisting of: varirubin (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Talixicizumab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utoluumab (PF-05082566), and Ureluumab (BMS-663513).

[0202] In some embodiments, the antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

[0203] In some embodiments, the TNFRSF member protein is TNFR2. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 as shown in Table 4. In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the sequences shown in Table 5.

[0204] In some embodiments, the antibody or its antigen-binding fragment is neither an agonist nor an antagonist of the TNFRSF member protein. In some embodiments, the TNFRSF member protein is TNFR2. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 as shown in Table 6. In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the sequences shown in Table 7.

[0205] In some embodiments, the antibody or its antigen-binding fragment has less than about 90% (e.g., less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10%) cross-reactivity with the soluble TNFRSF member protein. In some embodiments, the antibody or its antigen-binding fragment has less than 50% cross-reactivity with the soluble TNFRSF member protein.

[0206] In a tenth aspect, this disclosure is characterized by an agonist antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, said agonist antibody or antigen-binding fragment comprising:

[0207] (A) A component for binding to an epitope in human TNFR2, said component comprising:

[0208] (a) CDR-H1, wherein CDR-H1 comprises the following amino acid sequences:

[0209] (i)X 1 X 2 X2 X 1 X 2 X 3 X 2 JJJ, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X 2 Independently, it is Y, S, T, C, N, or Q; X 3 It is either D or E; and each J is independently a naturally occurring amino acid or is absent; or

[0210] (ii)X 1 X 2 X 2 X 3 X 1 , where each X 1 Independently, it is either D or E; each X 2 Independently, it is Y, S, T, C, N, or Q; and X 3 It is L, A, V, G, I, M, W, F, or P;

[0211] (b) CDR-H2, wherein CDR-H2 contains X 1 X 2 X 3 X 2 X 3 X 3 X 1 X 3 X 3 X 3 X 3 X 3 X 3 X 4 X 2 X 4 X 2 The amino acid sequence, wherein each X1 is independently D or E; each X2 is independently I, A, V, L, G, M, W, F or P; each X 3 Each of the following is independently N, S, T, C, Y, or Q; and each X4 is independently K, R, or H;

[0212] (c) CDR-H3, wherein CDR-H3 contains X 1 X 2 X 2 X 1 X 2 X 1 X 3 X 1 The amino acid sequence, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X 2 Independently, it is N, S, T, C, Y, or Q; and X3 It is either D or E;

[0213] (d) CDR-L1, wherein CDR-L1 includes X 1 X 2 X 1 X 1 X 1 X 2 X 3 X 4 X 1 X 1 The amino acid sequence, where each X 1 Independently, it is S, Y, T, C, N, or Q; each X 2 Independently, it is A, G, V, L, I, M, W, F, or P; X 3 It is R, H, or K; and X 4 It is Y, A, V, I, L, M, F, or W;

[0214] (e) CDR-L2, wherein the CDR-L2 includes X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is L, A, V, G, I, M, W, F, or P; and each X 2 Independently, it is T, S, C, Y, N, or Q; and / or

[0215] (f) CDR-L3, wherein the CDR-L3 includes X 1 X 2 X 2 X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is P, A, V, L, I, M, W, F, or G; and each X 2 Independently, it is Q, S, T, C, N, or Y; and

[0216] (B) An Fc domain, wherein the Fc domain comprises one or more amino acid substitutions, wherein:

[0217] (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0218] (i) L235E;

[0219] (ii) L234A and L235A;

[0220] (iii) L234A, L235A and P329G;

[0221] (iv) L234E, L235F and P331S;

[0222] (v)D265A;

[0223] (vi) G237A;

[0224] (vii) E318A;

[0225] (viii) E233P;

[0226] (ix) G236R and L328R;

[0227] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0228] (xi) N297G or N297Q;

[0229] (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0230] (i) D265A;

[0231] (ii) G237A;

[0232] (iii) E318A;

[0233] (iv) H268Q, V309L, A330S and P331S;

[0234] (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and

[0235] (vi) N297G or N297Q;

[0236] (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0237] (i) L235E;

[0238] (ii) L234A and L235A;

[0239] (iii) L234A, L235A and P329G;

[0240] (iv) L234E, L235F and P331S;

[0241] (v)D265A;

[0242] (vi) G237A;

[0243] (vii) E318A;

[0244] (viii) E233P;

[0245] (ix) G236R and L328R;

[0246] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0247] (xi) N297G or N297Q;

[0248] (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0249] (i) L235E;

[0250] (ii) D265A;

[0251] (iii) G237A;

[0252] (iv) E318A;

[0253] (v)E233P;

[0254] (vi) G236R and L328R; and

[0255] (vii) N297G or N297Q; or

[0256] (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P (according to Kabat's S241P), the Fc domain being a human IgG4 Fc domain, and optionally the one or more amino acid substitutions being selected from the group consisting of:

[0257] (i) L235E; and

[0258] (ii) F234A and L235A,

[0259] Furthermore, the amino acid positions are based on EU index numbers.

[0260] In some embodiments, the antibody or its antigen-binding fragment is an IgG4 antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment contains an amino acid substitution for S228P.

[0261] In some embodiments, the CDR-H1 includes GYTFTZ 1 Z 2 The amino acid sequence of JJJ (SEQ ID NO: 1480), where Z 1 It is D or T; Z 2 It is Y, F, or L; and each J is independently a naturally occurring amino acid or is absent. In some embodiments, the CDR-H1 comprises the amino acid sequence GYTFTDY (SEQ ID NO: 1413). In some embodiments, the antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0262] In an eleventh aspect, this disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to human TNFSF member proteins, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, further wherein:

[0263] (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0264] (i) L235E;

[0265] (ii) L234A and L235A;

[0266] (iii) L234A, L235A and P329G;

[0267] (iv) L234E, L235F and P331S;

[0268] (v)D265A;

[0269] (vi) G237A;

[0270] (vii) E318A;

[0271] (viii) E233P;

[0272] (ix) G236R and L328R;

[0273] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0274] (xi) N297G or N297Q;

[0275] (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0276] (i) D265A;

[0277] (ii) G237A;

[0278] (iii) E318A;

[0279] (iv) H268Q, V309L, A330S and P331S;

[0280] (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and

[0281] (vi) N297G or N297Q;

[0282] (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0283] (i) L235E;

[0284] (ii) L234A and L235A;

[0285] (iii) L234A, L235A and P329G;

[0286] (iv) L234E, L235F and P331S;

[0287] (v)D265A;

[0288] (vi) G237A;

[0289] (vii) E318A;

[0290] (viii) E233P;

[0291] (ix) G236R and L328R;

[0292] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0293] (xi) N297G or N297Q;

[0294] (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0295] (i) L235E;

[0296] (ii) D265A;

[0297] (iii) G237A;

[0298] (iv) E318A;

[0299] (v)E233P;

[0300] (vi) G236R and L328R; and

[0301] (vii) N297G or N297Q; or

[0302] (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P (according to Kabat's S241P), the Fc domain being a human IgG4 Fc domain, and optionally the one or more amino acid substitutions being selected from the group consisting of:

[0303] (i) L235E; and

[0304] (ii) F234A and L235A,

[0305] Furthermore, the amino acid positions are based on EU index numbers.

[0306] In some embodiments, the antibody or its antigen-binding fragment is an IgG4 antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment contains an amino acid substitution for S228P.

[0307] In some implementations, the TNFSF member protein is TRAIL.

[0308] In a twelfth aspect, this disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to human CD28, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, further wherein:

[0309] (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0310] (i) L235E;

[0311] (ii) L234A and L235A;

[0312] (iii) L234A, L235A and P329G;

[0313] (iv) L234E, L235F and P331S;

[0314] (v)D265A;

[0315] (vi) G237A;

[0316] (vii) E318A;

[0317] (viii) E233P;

[0318] (ix) G236R and L328R;

[0319] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0320] (xi) N297G or N297Q;

[0321] (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0322] (i) D265A;

[0323] (ii) G237A;

[0324] (iii) E318A;

[0325] (iv) H268Q, V309L, A330S and P331S;

[0326] (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and

[0327] (vi) N297G or N297Q;

[0328] (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0329] (i) L235E;

[0330] (ii) L234A and L235A;

[0331] (iii) L234A, L235A and P329G;

[0332] (iv) L234E, L235F and P331S;

[0333] (v)D265A;

[0334] (vi) G237A;

[0335] (vii) E318A;

[0336] (viii) E233P;

[0337] (ix) G236R and L328R;

[0338] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0339] (xi) N297G or N297Q;

[0340] (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0341] (i) L235E;

[0342] (ii) D265A;

[0343] (iii) G237A;

[0344] (iv) E318A;

[0345] (v)E233P;

[0346] (vi) G236R and L328R; and

[0347] (vii) N297G or N297Q; or

[0348] (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P (according to Kabat's S241P), the Fc domain being a human IgG4 Fc domain, and optionally the one or more amino acid substitutions being selected from the group consisting of:

[0349] (i) L235E; and

[0350] (ii) F234A and L235A,

[0351] Furthermore, the amino acid positions are based on EU index numbers.

[0352] In some embodiments, the antibody or its antigen-binding fragment is an IgG4 antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment contains an amino acid substitution for S228P.

[0353] In some embodiments, the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to CD28. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of cilalizumab (TAB-08). In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and VL sequences of cilalizumab (TAB-08).

[0354] In a thirteenth aspect, this disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to human ICOS, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, further wherein:

[0355] (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0356] (i) L235E;

[0357] (ii) L234A and L235A;

[0358] (iii) L234A, L235A and P329G;

[0359] (iv) L234E, L235F and P331S;

[0360] (v)D265A;

[0361] (vi) G237A;

[0362] (vii) E318A;

[0363] (viii) E233P;

[0364] (ix) G236R and L328R;

[0365] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0366] (xi) N297G or N297Q;

[0367] (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0368] (i) D265A;

[0369] (ii) G237A;

[0370] (iii) E318A;

[0371] (iv) H268Q, V309L, A330S and P331S;

[0372] (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and

[0373] (vi) N297G or N297Q;

[0374] (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0375] (i) L235E;

[0376] (ii) L234A and L235A;

[0377] (iii) L234A, L235A and P329G;

[0378] (iv) L234E, L235F and P331S;

[0379] (v)D265A;

[0380] (vi) G237A;

[0381] (vii) E318A;

[0382] (viii) E233P;

[0383] (ix) G236R and L328R;

[0384] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0385] (xi) N297G or N297Q;

[0386] (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of:

[0387] (i) L235E;

[0388] (ii) D265A;

[0389] (iii) G237A;

[0390] (iv) E318A;

[0391] (v)E233P;

[0392] (vi) G236R and L328R; and

[0393] (vii) N297G or N297Q; or

[0394] (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P (according to Kabat's S241P), the Fc domain being a human IgG4 Fc domain, and optionally the one or more amino acid substitutions being selected from the group consisting of:

[0395] (i) L235E; and

[0396] (ii) F234A and L235A,

[0397] Furthermore, the amino acid positions are based on EU index numbers.

[0398] In some embodiments, the antibody or its antigen-binding fragment is an IgG4 antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment contains an amino acid substitution for S228P.

[0399] In some embodiments, the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to ICOS. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of GSK-3359609 or JTX-2011. In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and VL sequences of GSK-3359609 or JTX-2011.

[0400] In some embodiments, the antibody or its antigen-binding fragment does not contain any of the amino acid substitutions shown in Table 18, and the amino acid positions are based on EU index numbers.

[0401] In some embodiments, the antibody or its antigen-binding fragment is selected from the group consisting of: monoclonal antibodies or their antigen-binding fragments, polyclonal antibodies or their antigen-binding fragments, human antibodies or their antigen-binding fragments, humanized antibodies or their antigen-binding fragments, primate-derived antibodies or their antigen-binding fragments, bispecific antibodies or their antigen-binding fragments, multispecific antibodies or their antigen-binding fragments, bivariate immunoglobulin domains, monovalent antibodies or their antigen-binding fragments, and chimeric antibodies or their antigen-binding fragments. In some embodiments, the antibody or its antigen-binding fragment is a human antibody, a humanized antibody, or a chimeric antibody or their antigen-binding fragments.

[0402] In a fourteenth aspect, the present disclosure is characterized by an antibody or an antigen-binding fragment thereof that specifically binds to a human TNFRSF member protein, a human TNFSF member protein, a human CD28 or a human ICOS, wherein the antibody or the antigen-binding fragment thereof does not contain an Fc domain.

[0403] In some embodiments, the TNFRSF member protein is selected from the group consisting of: TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3. In some embodiments, the TNFRSF member protein is TNFR2.

[0404] In some embodiments, the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to TNFR2. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 as shown in Table 2. In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and the VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the sequences shown in Table 3.

[0405] In some embodiments, the antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to TNFR2. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 as shown in Table 4. In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and the VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the sequences shown in Table 5.

[0406] In some implementations, the TNFSF member protein is TRAIL.

[0407] In some embodiments, the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to CD28. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of cilalizumab (TAB-08). In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and VL sequences of cilalizumab (TAB-08).

[0408] In some embodiments, the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to ICOS. In some embodiments, the antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of GSK-3359609 or JTX-2011. In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and VL sequences of GSK-3359609 or JTX-2011.

[0409] In some embodiments, the antibody or its antigen-binding fragment is selected from the group consisting of: single-chain Fv molecules (scFv), bifunctional antibodies, trifunctional antibodies, antibody-like protein scaffolds, Fv fragments, Fab fragments, F(ab')2 molecules, and tandem scFv (taFv).

[0410] In a fifteenth aspect, the present disclosure is characterized by a method for producing an antibody or an antigen-binding fragment thereof disclosed in the ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspects of the present disclosure, said method comprising expressing a polynucleotide encoding said antibody or an antigen-binding fragment thereof in a host cell and recovering said antibody or an antigen-binding fragment thereof from a host cell culture medium.

[0411] In a sixteenth aspect, the present disclosure is characterized by a construct comprising a first polypeptide domain and a second polypeptide domain, wherein the first polypeptide domain and the second polypeptide domain are each independently an antigen-binding fragment disclosed in the ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspects of the present disclosure.

[0412] In some embodiments, the first polypeptide domain and the second polypeptide domain are linked by a covalent linker. In some embodiments, the covalent linker comprises an amide bond or a disulfide bond.

[0413] In a seventeenth aspect, the present disclosure is characterized by a polynucleotide encoding an antibody or antigen-binding fragment thereof disclosed in the ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspects of the present disclosure, or a construct disclosed in the sixteenth aspect of the present disclosure.

[0414] In an eighteenth aspect, the present disclosure is characterized by a carrier comprising the polynucleotides disclosed in the seventeenth aspect of the present disclosure.

[0415] In some embodiments, the vector is an expression vector. In some embodiments, the expression vector is a eukaryotic expression vector.

[0416] In some embodiments, the vector is a viral vector. In some embodiments, the viral vector is selected from the group consisting of adenovirus, retrovirus, poxvirus, adeno-associated virus, baculovirus, herpes simplex virus, and vaccinia virus. In some embodiments, the adenovirus is serotype 1-60 adenovirus. In some embodiments, the adenovirus is serotype 5, 26, 35, or 48 adenovirus. In some embodiments, the retrovirus is gamma retrovirus or lentivirus. In some embodiments, the vaccinia virus is modified Ankara vaccinia (MVA).

[0417] In a nineteenth aspect, the present disclosure is characterized by isolated host cells comprising the vector disclosed in an eighteenth aspect of the present disclosure.

[0418] In some embodiments, the host cell is a prokaryotic or eukaryotic cell. In some embodiments, the eukaryotic cell is a mammalian cell. In some embodiments, the mammalian cell is a CHO cell.

[0419] In a twentieth aspect, the invention is characterized by a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof disclosed in the ninth, tenth, eleventh, twelfth, thirteenth, or fourteenth aspects of this disclosure, a construct disclosed in the sixteenth aspect of this disclosure, a polynucleotide disclosed in the seventeenth aspect of this disclosure, a vector disclosed in the eighteenth aspect of this disclosure, or a host cell disclosed in the nineteenth aspect of this disclosure, and a pharmaceutically acceptable carrier or excipient.

[0420] In a twentieth aspect, the present disclosure is characterized by a method for modulating the immune response of a human subject, the method comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed in the ninth, tenth, eleventh, twelfth, thirteenth, or fourteenth aspects of the present disclosure, a construct disclosed in the sixteenth aspect of the present disclosure, a polynucleotide disclosed in the seventeenth aspect of the present disclosure, a vector disclosed in the eighteenth aspect of the present disclosure, a host cell disclosed in the nineteenth aspect of the present disclosure, or a pharmaceutical composition disclosed in the twentieth aspect of the present disclosure.

[0421] In some embodiments, the method is used to treat the human subject with a cell proliferation disorder, autoimmune disease, infectious disease, inflammatory disease, neurological disease, allergy, transplant rejection (e.g., allogeneic graft rejection), or graft-versus-host disease. In some embodiments, the method is used for transplantation or for regeneration of tissues or organs of the human subject.

[0422] In some embodiments, the cell proliferation disorder is cancer. In some embodiments, the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to a TNFRSF member protein selected from the group consisting of: TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TNFR1, Fas, CD40, CD27, 4-1BB, OX40, GITR, and XEDAR. In some embodiments, the antibody or its antigen-binding fragment is an antagonist antibody or its antigen-binding fragment that specifically binds to a TNFRSF member protein selected from the group consisting of: TRAMP, NGFR, TRAIL-R4, TNFR2, HVEM, CD30, TROY, and RELT.

[0423] In some embodiments, the serum or plasma level of the soluble TNFRSF member protein in the human subject is lower than or the same as a reference level for the soluble TNFRSF member protein, and the method comprises administering to the subject an agonist antibody or an antigen-binding fragment thereof that specifically binds to the TNFRSF member protein. In some embodiments, the serum or plasma level of the soluble TNFRSF member protein in the human subject is higher than a reference level for the soluble TNFRSF member protein, and the method comprises administering to the subject an antagonist antibody or an antigen-binding fragment thereof that specifically binds to the TNFRSF member protein.

[0424] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to a TNFRSF member protein of this disclosure is administered to a human subject at a low dose. In some cases, the low dose is a dose level lower than, for example, the typical (or art-recognized) dose level of the corresponding antibody or antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment thereof is administered once weekly at a dose of about 0.8 mg / kg to about 1.2 mg / kg (e.g., about 0.8 mg / kg, about 0.85 mg / kg, about 0.9 mg / kg, about 0.95 mg / kg, about 1.0 mg / kg, about 1.05 mg / kg, about 1.1 mg / kg, about 1.15 mg / kg, or about 1.2 mg / kg). In some embodiments, the antibody or its antigen-binding fragment is administered every two weeks at a dose of about 1.65 mg / kg to about 2.5 mg / kg (e.g., about 1.65 mg / kg, about 1.7 mg / kg, about 1.75 mg / kg, about 1.8 mg / kg, about 1.85 mg / kg, about 1.9 mg / kg, about 1.95 mg / kg, about 2.0 mg / kg, about 2.05 mg / kg, about 2.1 mg / kg, about 2.15 mg / kg, about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg, or about 2.5 mg / kg). In some embodiments, the antibody or its antigen-binding fragment is administered once every three weeks at a dose of about 2.2 mg / kg to about 3.0 mg / kg (e.g., about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg, about 2.5 mg / kg, about 2.55 mg / kg, about 2.6 mg / kg, about 2.65 mg / kg, about 2.7 mg / kg, about 2.75 mg / kg, about 2.8 mg / kg, about 2.85 mg / kg, about 2.9 mg / kg, about 2.95 mg / kg, or about 3.0 mg / kg). In some embodiments, the antibody or its antigen-binding fragment is administered once every four weeks at a dose of about 2.5 mg / kg to about 4.2 mg / kg (e.g., about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4.0 mg / kg, about 4.1 mg / kg, or about 4.2 mg / kg).

[0425] In a twentieth aspect, the present disclosure is characterized in a kit comprising an agent selected from the group consisting of: an antibody or antigen-binding fragment thereof disclosed in the ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspects of the present disclosure, a construct disclosed in the sixteenth aspect of the present disclosure, a polynucleotide disclosed in the seventeenth aspect of the present disclosure, a vector disclosed in the eighteenth aspect of the present disclosure, a host cell disclosed in the nineteenth aspect of the present disclosure or a pharmaceutical composition disclosed in the twentieth aspect of the present disclosure.

[0426] In some embodiments, the kit further includes instructions for transfecting the vector into host cells. In some embodiments, the kit further includes instructions for expressing the antibody or its antigen-binding fragment or the construct in host cells. In some embodiments, the kit further includes reagents that can be used to express the antibody or its antigen-binding fragment or the construct in host cells. In some embodiments, the kit further includes instructions for administering the agent to a human subject. In some embodiments, the kit further includes instructions for preparing or using the agent. In some embodiments, the kit further includes instructions for determining the level of the soluble TNFRSF member protein.

[0427] In a twenty-third aspect, this disclosure is characterized by a method comprising:

[0428] (a)

[0429] (i) Detecting the level of soluble TNFRSF member proteins in a biofluid from a subject, wherein the level of the soluble TNFRSF member proteins in the biofluid is higher (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) than a reference level of the soluble TNFRSF member proteins; and

[0430] (ii) Administering to the subject at a predetermined dosing frequency one or more doses of an antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein, wherein the method is used to reduce the level of the soluble TNFRSF member protein; or

[0431] (b)

[0432] (i) Detecting the level of soluble TNFRSF member proteins in a biofluid from a subject, wherein the level of the soluble TNFRSF member proteins in the biofluid is normal or lower (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more lower) relative to a reference level of the soluble TNFRSF member proteins; and

[0433] (ii) Administering to the subject one or more doses of an agonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein at a given dosing frequency, wherein the method is used to increase the level of the soluble TNFRSF member protein.

[0434] In some embodiments, the normal level of the soluble TNFRSF member protein in the biofluid is the level of the soluble TNFRSF member protein in the biofluid (e.g., serum or plasma) of a healthy person. In some embodiments, the normal level of soluble TNFR2 in the serum of a human subject is about 1.53 ng / ml to about 3.00 ng / ml (e.g., about 1.80 ng / ml to about 3.00 ng / ml, about 2.10 ng / ml to about 3.00 ng / ml, about 2.40 ng / ml to about 3.00 ng / ml, about 2.70 ng / ml to about 3.00 ng / ml, about 1.53 ng / ml to about 2.70 ng / ml, about 1.80 ng / ml to about 2.70 ng / ml, about 2.10 ng / ml to about 2.70 ng / ml, about 2.40 ng / ml to about 3.0 ... 0 ng / ml to about 2.70 ng / ml, about 1.53 ng / ml to about 2.40 ng / ml, about 1.80 ng / ml to about 2.40 ng / ml, about 2.10 ng / ml to about 2.40 ng / ml, about 1.53 ng / ml to about 2.10 ng / ml, about 1.80 ng / ml to about 2.10 ng / ml, about 1.53 ng / ml to about 1.80 ng / ml, about 1.53 ng / ml, about 1.80 ng / ml, about 2.10 ng / ml, about 2.40 ng / ml, about 2.70 ng / ml or about 3.00 ng / ml). In some embodiments, the enhanced level of the soluble TNFRSF member protein in the biofluid is higher than the normal level of the soluble TNFRSF member protein in the biofluid (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more).In some implementations, the enhanced level of soluble TNFR2 in the serum of human subjects is from about 2.53 ng / ml to about 10 ng / ml (e.g., about 2.53 ng / ml to about 3 ng / ml, about 2.53 ng to about 4 ng / ml, about 2.53 ng to about 6 ng / ml, about 2.53 ng to about 8 ng / ml, about 3 ng / ml to about 4 ng / ml, about 3 ng / ml to about 6 ng / ml, about 3 ng / ml to about 8 ng / ml, about 3 ng / ml to about 10 ng / ml, about 4 ng / ml to about 6 ng / ml, about 4 ng / ml to about 8 ng / ml, about 4 ng / ml to about 10 ng / ml, about 6 ng / ml to about 8 ng / ml, about 6 ng / ml to about 10 ng / ml, about 8 ng / ml to about 10 ng / ml, about 2.53 ng / ml, about 3 ng / ml, about 4 ng / ml, about 6 ng / ml, about 8 ng / ml, or about 10 ng / ml). When the level of the soluble TNFRSF member protein in the human subject is higher than the normal level of the soluble TNFRSF member protein (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more), an antagonistic antibody or its antigen-binding fragment that specifically binds to the corresponding TNFRSF member protein may be used to reduce the level of the soluble TNFRSF member protein in the human subject (e.g., reduce it to the normal level of the soluble TNFRSF member protein).

[0435] In some embodiments, a decrease in the level of the soluble TNFRSF member protein in the human subject (e.g., a decrease of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or more) indicates insufficient T-cell suppression and a need for greater TNFR2 function, such as in cases of cell proliferation disorders, autoimmune diseases, infectious diseases, inflammatory diseases, neurological diseases, allergies, transplant rejection (e.g., allogeneic graft rejection), or graft-versus-host disease in the human subject. In some embodiments, the neurological disease is Alzheimer's disease. In this case, the level of the soluble TNFRSF member protein can be increased using a corresponding TNFRSF agonist (and an agonistic antibody or antigen-binding fragment thereof that binds to the TNFRSF member protein corresponding to the soluble TNFRSF member protein, as disclosed herein). This strategy can be used to screen antibodies, determine optimal dosing levels of TNFRSF agonists in clinical trials, or ensure that dosing levels of TNFRSF antagonists administered to subjects are therapeutically effective in clinical practice. For example, decreased soluble TNFR2 levels in human subjects can indicate an outbreak of autoimmunity or impending transplant rejection. Increased soluble TNFR2 levels in human subjects can indicate that cancers with Treg and / or TNFR2-driven mechanisms require therapy to reduce soluble TNFR2 levels (e.g., by eliminating suppressor cells, such as Treg cells or MDSCs). In organ or tissue transplantation, the host may have normal levels of soluble TNFR2, but it is desirable to increase soluble TNFR2 levels (e.g., by administering a TNFR2 agonist to the host, such as the TNFR2-binding agonist antibody described herein) to maintain the viability of the transplanted organ or tissue by promoting Treg cell activity.

[0436] In some embodiments, the method includes detecting a reference level of the soluble TNFRSF member protein, a normal level of the soluble TNFRSF member protein in the subject's biofluid, or an elevated level of the soluble TNFRSF member protein (e.g., an increase of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or more) of the soluble TNFRSF member protein, wherein the method further includes altering (e.g., increasing or decreasing) the dose and / or frequency of administration of the antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein. In some embodiments, the antagonistic antibody or its antigen-binding fragment that specifically binds to the soluble TNFRSF member protein reduces or increases (e.g., reduces or increases by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) the level of the TNFRSF member protein. In a preferred embodiment, the antagonistic antibody or its antigen-binding fragment that specifically binds to the soluble TNFRSF member protein reduces (e.g., reduces by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) the level of the TNFRSF member protein.

[0437] In some embodiments, the method includes detecting a reference level of the soluble TNFRSF member protein, a normal level of the soluble TNFRSF member protein in the subject's biofluid, or a reduced level of the soluble TNFRSF member protein (e.g., a reduction of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or more) of the soluble TNFRSF member protein, wherein the method further includes altering (e.g., increasing or decreasing) the dose and / or frequency of administration of the agonist antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein. In some embodiments, the agonist antibody or its antigen-binding fragment that specifically binds to the soluble TNFRSF member protein reduces or increases (e.g., reduces or increases by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) the level of the TNFRSF member protein. In a preferred embodiment, the agonist antibody or its antigen-binding fragment that specifically binds to the soluble TNFRSF member protein increases (e.g., increases by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) the level of the TNFRSF member protein.

[0438] In some embodiments, the method includes detecting an elevated level (e.g., an increase of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) of the soluble TNFRSF member protein in the subject's biofluid relative to a reference level, wherein the method further includes:

[0439] (a) Increasing the dose and / or frequency of administration of the antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein; or

[0440] (b) Reduce the dose and / or frequency of the agonist antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

[0441] In some embodiments, the method includes detecting a normal or reduced (e.g., reduced by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) level of the soluble TNFRSF member protein in the subject's biofluid relative to a reference level, wherein the method further includes:

[0442] (a) Increasing the dose and / or frequency of administration of the agonist antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein; or

[0443] (b) Reduce the dose and / or frequency of administration of the antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

[0444] In some embodiments, the TNFRSF member protein is selected from the group consisting of: TNFR2, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TNFR1, TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3. In some embodiments, the TNFRSF member protein is TNFR2.

[0445] In some embodiments, the antagonistic antibody or the agonist antibody or their antigen-binding fragments that specifically bind to the TNFRSF member protein are administered to the human subject at a dosing frequency of once weekly, once every two weeks, once every three weeks, or once every four weeks.

[0446] In some embodiments, the antagonistic antibody or the agonistic antibody or their antigen-binding fragments that specifically bind to the TNFRSF member proteins are present at a concentration of about 0.8 mg / kg to about 1.2 mg / kg (e.g., about 0.8 mg / kg, about 0.85 mg / kg, about 0.9 mg / kg, about 0.95 mg / kg, about 1.0 mg / kg, about 1.05 mg / kg, about 1.1 mg / kg, about 1.15 mg / kg, or about 1.2 mg / kg), about 1.65 mg / kg to about 2.5 mg / kg (e.g., about 1.65 mg / kg). mg / kg, approximately 1.7 mg / kg, approximately 1.75 mg / kg, approximately 1.8 mg / kg, approximately 1.85 mg / kg, approximately 1.9 mg / kg, approximately 1.95 mg / kg, approximately 2.0 mg / kg, approximately 2.05 mg / kg, approximately 2.1 mg / kg, approximately 2.15 mg / kg, approximately 2.2 mg / kg, approximately 2.25 mg / kg, approximately 2.3 mg / kg, approximately 2.35 mg / kg, approximately 2.4 mg / kg, approximately 2.45 mg / kg or approximately 2.5 mg / kg), approximately 2.2 mg / kg to approximately 3.0 mg / kg. / kg (e.g., about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg, about 2.5 mg / kg, about 2.55 mg / kg, about 2.6 mg / kg, about 2.65 mg / kg, about 2.7 mg / kg, about 2.75 mg / kg, about 2.8 mg / kg, about 2.85 mg / kg, about 2.9 mg / kg, about 2.95 mg / kg, or about 3.0 mg / kg) or about 2.5 mg / kg to about 4.2 mg / kg. The human subjects were given doses of g / kg (e.g., about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4.0 mg / kg, about 4.1 mg / kg, or about 4.2 mg / kg).

[0447] In some embodiments, the antagonistic antibody or the agonistic antibody, or their antigen-binding fragments, that specifically bind to the TNFRSF member protein are administered to the human subject in the following manner:

[0448] (a) Once a week at a dose of about 0.8 mg / kg to about 1.2 mg / kg (e.g., about 0.8 mg / kg, about 0.85 mg / kg, about 0.9 mg / kg, about 0.95 mg / kg, about 1.0 mg / kg, about 1.05 mg / kg, about 1.1 mg / kg, about 1.15 mg / kg or about 1.2 mg / kg);

[0449] (b) Every two weeks at a dose of about 1.65 mg / kg to about 2.5 mg / kg (e.g., about 1.65 mg / kg, about 1.7 mg / kg, about 1.75 mg / kg, about 1.8 mg / kg, about 1.85 mg / kg, about 1.9 mg / kg, about 1.95 mg / kg, about 2.0 mg / kg, about 2.05 mg / kg, about 2.1 mg / kg, about 2.15 mg / kg, about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg or about 2.5 mg / kg);

[0450] (c) Every three weeks at a dose of about 2.2 mg / kg to about 3.0 mg / kg (e.g., about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg, about 2.5 mg / kg, about 2.55 mg / kg, about 2.6 mg / kg, about 2.65 mg / kg, about 2.7 mg / kg, about 2.75 mg / kg, about 2.8 mg / kg, about 2.85 mg / kg, about 2.9 mg / kg, about 2.95 mg / kg, or about 3.0 mg / kg); or

[0451] (d) Administer at a dose of about 2.5 mg / kg to about 4.2 mg / kg (e.g., about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4.0 mg / kg, about 4.1 mg / kg or about 4.2 mg / kg) once every four weeks.

[0452] In some embodiments, the method further comprises reducing the dose range of the antagonistic antibody or the agonistic antibody or their antigen-binding fragments that specifically bind to the TNFRSF member protein.

[0453] In some embodiments, the antagonistic antibody or the agonist antibody, or their antigen-binding fragments, that specifically bind to the TNFRSF member protein are administered subcutaneously to the human subject. In some embodiments, the level of soluble TNFR2 in the human subject is the serum or plasma level of soluble TNFR2 in the human subject. In some embodiments, the burden (e.g., cell number) of Treg cells or MDSCs (e.g., Treg cells or MDSCs expressing TNFR2) is measured directly in the tumor microenvironment of a human subject with cancer.

[0454] In some embodiments, the method is used to treat the human subject with a cell proliferation disorder, autoimmune disease, infectious disease, inflammatory disease, neurological disease, allergy, transplant rejection (e.g., allogeneic graft rejection), or graft-versus-host disease. In some embodiments, the method is used for transplantation or for regeneration of tissues or organs of the human subject. In some embodiments, the method is used to treat a cell proliferation disorder, and the antibody or its antigen-binding fragment is an antagonist antibody or its antigen-binding fragment that specifically binds to: TRAMP, DR6, NGFR, RANK, FN14, TNFR2, LT-β receptor, HVEM, CD30, BCMA, TACI, BAFF, TROY, or RELT. In some embodiments, the method is used to treat a cell proliferation disorder, and the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to: TRAIL-R1, TRAIL-R2, TRAIL-R3, TRAIL-R4, TNFR1, Fas, CD40, CD27, 4-1BB, OX40, GITR, or XEDAR. In some embodiments, the method is used to treat autoimmune diseases, and the antibody or its antigen-binding fragment is an antagonist antibody or its antigen-binding fragment that specifically binds to: TRAIL-R1, TRAIL-R2, TRAIL-R3, TRAIL-R4, TNFR1, CD40, CD27, 4-1BB, OX40, GITR, or XEDAR. In some embodiments, the method is used to treat autoimmune diseases, and the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to: TRAMP, DR6, NGFR, FN14, TNFR2, LT-β receptor, HVEM, CD30, or RELT. In some embodiments, the method is used to treat infectious diseases, and the antibody or its antigen-binding fragment is an antagonist antibody or its antigen-binding fragment that specifically binds to: TRAMP, DR6, TNFR2, HVEM, or RELT. In some embodiments, the method is used to treat infectious diseases, and the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to the following: TRAIL-R1, TRAIL-R2, TRAIL-R3, TRAIL-R4, CD40, CD27, 4-1BB, OX40, BCMA, TACI, or BAFF. In some embodiments, the method is used to treat inflammatory diseases, and the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to the following: DR6, FN14, TNFR2, HVEM, or RELT.In some embodiments, the method is used to treat neurological disorders, and the antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to DR6. In some embodiments, the method is used to treat neurological disorders (e.g., CNS diseases), and the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to TNFR2. In some embodiments, the method is used to treat allergies, and the antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to TNFR1, CD40, CD27, 4-1BB, OX40, GITR, or XEDAR. In some embodiments, the method is used to treat transplant rejection (e.g., allogeneic graft rejection) or graft-versus-host disease, and the antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to TNFR1, 4-1BB, OX40, GITR, or XEDAR. In some embodiments, the method is used to treat transplant rejection (e.g., allogeneic graft rejection) or graft-versus-host disease, and the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to CD30 or TNFR2.

[0455] In some implementations, the biofluid is serum or plasma.

[0456] In some embodiments, the reference level of the soluble TNFRSF member protein is the level of the soluble TNFRSF in the biofluid of a healthy person.

[0457] In some embodiments, one or more doses of the antibody or its antigen-binding fragment are administered to the human subject during one or more treatment periods, wherein:

[0458] (a) Each dose contains:

[0459] (i) about 0.1 mg to about 5000 mg of the antibody or its antigen-binding fragment; or

[0460] (ii) the antibody or its antigen-binding fragment at a concentration of about 0.001 mg / kg to about 50 mg / kg; and / or

[0461] (b) Each treatment period lasts for one, two, three, four, five, six, seven, eight or longer weeks.

[0462] In some embodiments, the dosing frequency is once or more per month, every three weeks, every two weeks, weekly, every six days, every five days, every four days, every three days, every two days, or daily. In some embodiments, the dosing frequency is once a week (Q1W), once every two weeks (Q2W), once every three weeks (Q3W), or once every four weeks (Q4W).

[0463] In some embodiments, the antagonistic antibody or the agonist antibody or their antigen-binding fragments that specifically bind to the TNFRSF member protein are administered subcutaneously to the human subject.

[0464] In a twenty-fourth aspect, this disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3, and further wherein:

[0465] (a) The CDR-H1 contains the amino acid sequence GXTFXXY (SEQ ID NO: 1449), wherein each X is independently a naturally occurring amino acid;

[0466] (b) The CDR-H2 comprises the amino acid sequence of NPNYDS (SEQ ID NO: 1414), NYDSTS (SEQ ID NO: 1420) or variants thereof having at most two conserved amino acid substitutions;

[0467] (c) The CDR-H3 comprises the amino acid sequence of GNSWYFDV (SEQ ID NO: 1415) or a variant thereof having at most two conserved amino acid substitutions;

[0468] (d) The CDR-L1 comprises the amino acid sequence of SASSSVRYMY (SEQ ID NO: 1416) or a variant thereof having at most two conserved amino acid substitutions;

[0469] (e) The CDR-L2 comprises the amino acid sequence of LTSNLAS (SEQ ID NO: 1417) or a variant thereof having at most two conserved amino acid substitutions; and

[0470] (f) The CDR-L3 comprises the amino acid sequence of QQWSSNPLT (SEQ ID NO: 1418) or a variant thereof having at most two conserved amino acid substitutions.

[0471] In some embodiments, the CDR-H1 comprises an amino acid sequence selected from the group consisting of: GYTFXXY (SEQ ID NO: 1450), GSTFXXY (SEQ ID NO: 1451), GTTFXXY (SEQ ID NO: 1452), GCTFXXY (SEQ ID NO: 1453), GNTFXXY (SEQ ID NO: 1454), GQTFXXY (SEQ ID NO: 1455), GXTFSXY (SEQ ID NO: 1456), GXTFCXY (SEQ ID NO: 1457), GXTFYXY (SEQ ID NO: 1458), GXTFNXY (SEQ ID NO: 1459), GXTFQXY (SEQ ID NO: 1460), and GXTFXDY (SEQ ID NO: 1461), wherein each X is independently a naturally occurring amino acid.

[0472] In some embodiments, the CDR-H1 comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413) or a variant thereof having at most two conserved amino acid substitutions.

[0473] In some embodiments, CDR-H1 contains the amino acid sequence GYTFTDY (SEQ ID NO: 1413), CDR-H2 contains the amino acid sequence NPNYDS (SEQ ID NO: 1414), CDR-H3 contains the amino acid sequence GNSWYFDV (SEQ ID NO: 1415), CDR-L1 contains the amino acid sequence SASSVRYMY (SEQ ID NO: 1416), CDR-L2 contains the amino acid sequence LTSNLAS (SEQ ID NO: 1417), and CDR-L3 contains the amino acid sequence QQWSSNPLT (SEQ ID NO: 1418).

[0474] In some embodiments, CDR-H1 contains the amino acid sequence GYTFTDY (SEQ ID NO: 1413), CDR-H2 contains the amino acid sequence NYDSTS (SEQ ID NO: 1420), CDR-H3 contains the amino acid sequence GNSWYFDV (SEQ ID NO: 1415), CDR-L1 contains the amino acid sequence SASSVRYMY (SEQ ID NO: 1416), CDR-L2 contains the amino acid sequence LTSNLAS (SEQ ID NO: 1417), and CDR-L3 contains the amino acid sequence QQWSSNPLT (SEQ ID NO: 1418).

[0475] In a twenty-fifth aspect, the present disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3, and further wherein:

[0476] (a) The CDR-H1 comprises the amino acid sequence of GYTFTDYNLD (SEQ ID NO: 1425), DYIMH (SEQ ID NO: 1426) or variants thereof having at most two conserved amino acid substitutions;

[0477] (b) The CDR-H2 comprises the amino acid sequence of DINPNYDS (SEQ ID NO: 1427), WVDPEYGSTDYAEKFKK (SEQ ID NO: 1428), or variants thereof having at most two conserved amino acid substitutions;

[0478] (c) The CDR-H3 comprises the amino acid sequence of TSYSQKFRG (SEQ ID NO: 1429), DDGSYSPFEDY (SEQ ID NO: 1430) or variants thereof having at most two conserved amino acid substitutions;

[0479] (d) The CDR-L1 comprises an amino acid sequence selected from the group consisting of: SASSSVRYMYWY (SEQ ID NO: 1431), KASENVVTYVS (SEQ ID NO: 1432), RSSQSLVHSNGNTYLH (SEQ ID NO: 1433), KASENVVTYVS (SEQ ID NO: 1434), QASQNINKYIA (SEQ ID NO: 1435), QNINKY (SEQ ID NO: 1436), and variants thereof having at most two conserved amino acid substitutions;

[0480] (e) The CDR-L2 comprises an amino acid sequence selected from the group consisting of: LTSNLAS (SEQ ID NO: 1437), VTSNLAS (SEQ ID NO: 1438), LTSNLGS (SEQ ID NO: 1439), GASNRTY (SEQ ID NO: 1440), IKVSNRFS (SEQ ID NO: 1441), DPEYGS (SEQ ID NO: 1442), YTSTLES (SEQ ID NO: 1443), YTS, and variants thereof having at most two conserved amino acid substitutions; and

[0481] (f) The CDR-L3 comprises an amino acid sequence selected from the group consisting of: QQRSNWP (SEQ ID NO: 1445), GQGYSYPYT (SEQ ID NO: 1446), SQTTHVPPT (SEQ ID NO: 1447), LQYVNLIT (SEQ ID NO: 1448), and variants thereof having at most two conserved amino acid substitutions.

[0482] In some embodiments, CDR-H1 comprises the amino acid sequence of GYTFTDYNLD (SEQ ID NO: 1425) or DYIMH (SEQ ID NO: 1426); CDR-H2 comprises the amino acid sequence of DINPNYDS (SEQ ID NO: 1427) or WVDPEYGSTDYAEKFKK (SEQ ID NO: 1428); CDR-H3 comprises the amino acid sequence of TSYSQKFRG (SEQ ID NO: 1429) or DDGSYSPFEDY (SEQ ID NO: 1430); and CDR-L1 comprises the amino acid sequence selected from the group consisting of: SASSVRYMYWY (SEQ ID NO: 1431), KASENVVTYVS (SEQ ID NO: 1432), RSSQSLVHSNGNTYLH (SEQ ID NO: 1433), KASENVVTYVS (SEQ ID NO: 1434), QASQNINKYIA (SEQ ID NO: 1426). CDR-L2 comprises amino acid sequences selected from the group consisting of: LTSNLAS (SEQ ID NO: 1435), VTSNLAS (SEQ ID NO: 1438), LTSNLGS (SEQ ID NO: 1439), GASNRTY (SEQ ID NO: 1440), IKVSNRFS (SEQ ID NO: 1441), DPEYGS (SEQ ID NO: 1442), YTSTLES (SEQ ID NO: 1443), and YTS; and CDR-L3 comprises amino acid sequences selected from the group consisting of: QQRSNWP (SEQ ID NO: 1445), GQGYSYPYT (SEQ ID NO: 1446), SQTTHVPPT (SEQ ID NO: 1447), and LQYVNLIT (SEQ ID NO: 1448).

[0483] In a twenty-sixth aspect, this disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, wherein said antibody or antigen-binding fragment comprises:

[0484] (a) VH, said VH having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to any of SEQ ID NO: 1486-1490; and

[0485] (b) VL, wherein the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to any of SEQ ID NO: 1492-1497.

[0486] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0487] (a) VH, said VH having the amino acid sequence of any one of SEQ ID NO: 1486-1490; and

[0488] (b) VL, wherein the VL has the amino acid sequence of any one of SEQ ID NO: 1492-1497.

[0489] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0490] (a) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1488, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1495;

[0491] (b) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1488, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1497;

[0492] (c) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1489, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1496;

[0493] (d) VH and VL, wherein VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100%) to SEQ ID NO: 1490, and VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100%) to SEQ ID NO: 1496; or

[0494] (e) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1490, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1497.

[0495] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0496] (a) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1488 and VL has the amino acid sequence of SEQ ID NO: 1495;

[0497] (b) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1488 and VL has the amino acid sequence of SEQ ID NO: 1497;

[0498] (c) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1489 and VL has the amino acid sequence of SEQ ID NO: 1496;

[0499] (d) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1490 and VL has the amino acid sequence of SEQ ID NO: 1496; or

[0500] (e) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1490 and VL has the amino acid sequence of SEQ ID NO: 1497.

[0501] In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1488, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1495.

[0502] In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH has the amino acid sequence of SEQ ID NO: 1488 and the VL has the amino acid sequence of SEQ ID NO: 1495.

[0503] In some embodiments, the antibody or its antigen-binding fragment is a human antibody or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment is an IgG antibody or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment is a human IgG1 antibody, a human IgG2 antibody, a human IgG3 antibody, or a human IgG4 antibody or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment is a human IgG4 antibody or its antigen-binding fragment.

[0504] In some embodiments, the antibody or its antigen-binding fragment further comprises a human IgG4 Fc domain. In some embodiments, the human IgG4 Fc domain comprises a human IgG4 hinge region having an amino acid substitution S228P according to EU index number (S241P according to Kabat). In some embodiments, the modified antibody or its antigen-binding fragment comprises an amino acid substitution S228P.

[0505] In a twenty-seventh aspect, this disclosure is characterized by an antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, wherein the antibody or antigen-binding fragment thereof comprises:

[0506] (a) a heavy chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to any of SEQ ID NO: 1498-1503; and

[0507] (b) A light chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to any of SEQ ID NO: 1504-1510.

[0508] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0509] (a) a heavy chain having the amino acid sequence of any one of SEQ ID NO: 1498-1503; and

[0510] (b) A light chain having an amino acid sequence of any one of SEQ ID NO: 1504-1510.

[0511] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0512] (a) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to that of SEQ ID NO: 1498, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to that of SEQ ID NO: 1504;

[0513] (b) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1501, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1508;

[0514] (c) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1501, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1510;

[0515] (d) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1502, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1509;

[0516] (e) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1503, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1509; or

[0517] (f) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1503, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1510.

[0518] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0519] (a) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1498 and the light chain has the amino acid sequence of SEQ ID NO: 1504;

[0520] (b) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1501 and the light chain has the amino acid sequence of SEQ ID NO: 1508;

[0521] (c) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1501 and the light chain has the amino acid sequence of SEQ ID NO: 1510;

[0522] (d) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1502 and the light chain has the amino acid sequence of SEQ ID NO: 1509;

[0523] (e) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1503 and the light chain has the amino acid sequence of SEQ ID NO: 1509; or

[0524] (f) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1503 and the light chain has the amino acid sequence of SEQ ID NO: 1510.

[0525] In some embodiments, the antibody or its antigen-binding fragment comprises a heavy chain and a light chain, the heavy chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1501, and the light chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1508.

[0526] In some embodiments, the antibody or its antigen-binding fragment comprises a heavy chain and a light chain, the heavy chain having the amino acid sequence of SEQ ID NO: 1501 and the light chain having the amino acid sequence of SEQ ID NO: 1508.

[0527] In some embodiments, the antibody or its antigen-binding fragment is a human IgG4 antibody or its antigen-binding fragment.

[0528] In some embodiments of aspects 24, 25, 26 or 27, the antibody or its antigen-binding fragment is an agonist antibody or its antigen-binding fragment that specifically binds to human TNFR2.

[0529] In some embodiments, the antibody or its antigen-binding fragment is selected from the group consisting of: monoclonal antibodies or their antigen-binding fragments, polyclonal antibodies or their antigen-binding fragments, human antibodies or their antigen-binding fragments, humanized antibodies or their antigen-binding fragments, primate-derived antibodies or their antigen-binding fragments, bispecific antibodies or their antigen-binding fragments, multispecific antibodies or their antigen-binding fragments, bivariate immunoglobulin domains, monovalent antibodies or their antigen-binding fragments, chimeric antibodies or their antigen-binding fragments, single-chain Fv molecules (scFv), bifunctional antibodies, trifunctional antibodies, antibody-like protein scaffolds, Fv fragments, Fab fragments, F(ab')2 molecules, and tandem scFv (taFv). In some embodiments, the antibody or its antigen-binding fragment is a human antibody, a humanized antibody, or a chimeric antibody or their antigen-binding fragments.

[0530] In a twentieth aspect, the present disclosure is characterized by a method for producing an antibody or an antigen-binding fragment thereof disclosed in the twentieth, twentieth, twentieth, or twentieth seven aspects of the present disclosure, the method comprising expressing a polynucleotide encoding the antibody or an antigen-binding fragment thereof in a host cell and recovering the antibody or an antigen-binding fragment thereof from a host cell culture medium.

[0531] In a twentieth aspect, the present disclosure is characterized by a construct comprising a first polypeptide domain and a second polypeptide domain, wherein the first polypeptide domain and the second polypeptide domain are each independently an antigen-binding fragment disclosed in the twentieth, twentieth, twentieth, or twentieth aspects of the present disclosure.

[0532] In some embodiments, the first polypeptide domain and the second polypeptide domain are linked by a covalent linker. In some embodiments, the covalent linker comprises an amide bond or a disulfide bond.

[0533] In the thirtieth aspect, the present disclosure is characterized by a polynucleotide encoding an antibody or antigen-binding fragment thereof disclosed in the twenty-fourth, twenty-fifth, twenty-sixth or twenty-seventh aspects of the present disclosure, or a construct disclosed in the twenty-seventh aspect of the present disclosure.

[0534] In a thirty-first aspect, the present disclosure is characterized by a carrier comprising the polynucleotides disclosed in the thirty-first aspect of the present disclosure.

[0535] In some embodiments, the vector is an expression vector. In some embodiments, the expression vector is a eukaryotic expression vector.

[0536] In some embodiments, the vector is a viral vector. In some embodiments, the viral vector is selected from the group consisting of adenovirus, retrovirus, poxvirus, adeno-associated virus, baculovirus, herpes simplex virus, and vaccinia virus. In some embodiments, the adenovirus is serotype 1-60 adenovirus. In some embodiments, the adenovirus is serotype 5, 26, 35, or 48 adenovirus. In some embodiments, the retrovirus is gamma retrovirus or lentivirus. In some embodiments, the vaccinia virus is modified Ankara vaccinia vaccine (MVA).

[0537] In a thirty-second aspect, the present disclosure is characterized by isolated host cells comprising the vector disclosed in a thirty-first aspect of the present disclosure.

[0538] In some embodiments, the host cell is a prokaryotic or eukaryotic cell. In some embodiments, the eukaryotic cell is a mammalian cell. In some embodiments, the mammalian cell is a CHO cell.

[0539] In a thirty-third aspect, the present invention is characterized by a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof disclosed in the twenty-fourth, twenty-fifth, twenty-sixth or twenty-seventh aspects of the present disclosure, a construct disclosed in the twenty-ninth aspect of the present disclosure, a polynucleotide disclosed in the thirtieth aspect of the present disclosure, a vector disclosed in the thirty-first aspect of the present disclosure or a host cell disclosed in the thirty-second aspect of the present disclosure, and a pharmaceutically acceptable carrier or excipient.

[0540] In a thirty-fourth aspect, the present disclosure is characterized by a method for modulating the immune response of a human subject, the method comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed in the twenty-fourth, twenty-fifth, twenty-sixth, or twenty-seventh aspects of the present disclosure, a construct disclosed in the twenty-ninth aspect of the present disclosure, a polynucleotide disclosed in the thirtieth aspect of the present disclosure, a vector disclosed in the thirty-first aspect of the present disclosure, a host cell disclosed in the thirty-second aspect of the present disclosure, or a pharmaceutical composition disclosed in the thirty-third aspect of the present disclosure.

[0541] In some embodiments, the method is used to treat the human subject with a cell proliferation disorder, autoimmune disease, infectious disease, inflammatory disease, neurological disease, allergy, transplant rejection (e.g., allogeneic graft rejection), or graft-versus-host disease. In some embodiments, the method is used for transplantation or for regeneration of tissues or organs of the human subject.

[0542] In some embodiments, the method is used to treat cancer. In some embodiments, the human subject's blood (e.g., serum or plasma) level of soluble TNFR2 is normal or lower than a reference level for soluble TNFR2. In some embodiments, the method is used to increase the serum or plasma level of soluble TNFR2 in the human subject.

[0543] In some implementations, the method further includes:

[0544] (a) Obtaining a biological fluid (e.g., blood, plasma, or serum) from the human subject.

[0545] (b) Detect the reduction (e.g., a reduction of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) of soluble TNFR2 in the subject's biofluid relative to a reference level; and

[0546] (c) Administer one or more doses of the antibody or its antigen-binding fragment to the subject at a certain dosing frequency.

[0547] In some implementations, the method further includes:

[0548] (a) Detecting normal or reduced (e.g., reduced by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more) levels of soluble TNFR2 in the subject, and thereby increasing the level of soluble TNFR2 by administering the antibody or its antigen-binding fragment at an increased dose and / or frequency; or

[0549] (b) Detect the level of increase in soluble TNFR2 in the subject relative to a reference level of soluble TNFR2 (e.g., an increase of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% or more), and increase the dose and / or frequency of administration of the antibody or its antigen-binding fragment.

[0550] In some embodiments, the antibody or its antigen-binding fragment is administered to the human subject at a dosing frequency of once weekly, once every two weeks, once every three weeks, or once every four weeks. In some embodiments, the antibody or its antigen-binding fragment is administered at a dose of about 0.8 mg / kg to about 1.2 mg / kg (e.g., about 0.8 mg / kg, about 0.85 mg / kg, about 0.9 mg / kg, about 0.95 mg / kg, about 1.0 mg / kg, about 1.05 mg / kg, about 1.1 mg / kg, about 1.15 mg / kg, or about 1.2 mg / kg), about 1.65 mg / kg to about 2.5 mg / kg (e.g., about 1.65 mg / kg, about 1.7 mg / kg, about 1.75 mg / kg, or about 1.75 mg / kg). kg, approximately 1.8 mg / kg, approximately 1.85 mg / kg, approximately 1.9 mg / kg, approximately 1.95 mg / kg, approximately 2.0 mg / kg, approximately 2.05 mg / kg, approximately 2.1 mg / kg, approximately 2.15 mg / kg, approximately 2.2 mg / kg, approximately 2.25 mg / kg, approximately 2.3 mg / kg, approximately 2.35 mg / kg, approximately 2.4 mg / kg, approximately 2.45 mg / kg or approximately 2.5 mg / kg), approximately 2.2 mg / kg to approximately 3.0 mg / kg (e.g., approximately 2.2 mg / kg). Approximately 2.25 mg / kg, approximately 2.3 mg / kg, approximately 2.35 mg / kg, approximately 2.4 mg / kg, approximately 2.45 mg / kg, approximately 2.5 mg / kg, approximately 2.55 mg / kg, approximately 2.6 mg / kg, approximately 2.65 mg / kg, approximately 2.7 mg / kg, approximately 2.75 mg / kg, approximately 2.8 mg / kg, approximately 2.85 mg / kg, approximately 2.9 mg / kg, approximately 2.95 mg / kg, or approximately 3.0 mg / kg) or approximately 2.5 mg / kg to approximately 4.2 mg / kg (e.g., The antibody or its antigen-binding fragment is administered to the human subject at doses of approximately 2.5 mg / kg, approximately 2.6 mg / kg, approximately 2.7 mg / kg, approximately 2.8 mg / kg, approximately 2.9 mg / kg, approximately 3.0 mg / kg, approximately 3.1 mg / kg, approximately 3.2 mg / kg, approximately 3.3 mg / kg, approximately 3.4 mg / kg, approximately 3.5 mg / kg, approximately 3.6 mg / kg, approximately 3.7 mg / kg, approximately 3.8 mg / kg, approximately 3.9 mg / kg, approximately 4.0 mg / kg, approximately 4.1 mg / kg, or approximately 4.2 mg / kg. In some embodiments, the antibody or its antigen-binding fragment is administered to the human subject in the following manner:

[0551] (a) Once a week at a dose of about 0.8 mg / kg to about 1.2 mg / kg (e.g., about 0.8 mg / kg, about 0.85 mg / kg, about 0.9 mg / kg, about 0.95 mg / kg, about 1.0 mg / kg, about 1.05 mg / kg, about 1.1 mg / kg, about 1.15 mg / kg or about 1.2 mg / kg);

[0552] (b) Every two weeks at a dose of about 1.65 mg / kg to about 2.5 mg / kg (e.g., about 1.65 mg / kg, about 1.7 mg / kg, about 1.75 mg / kg, about 1.8 mg / kg, about 1.85 mg / kg, about 1.9 mg / kg, about 1.95 mg / kg, about 2.0 mg / kg, about 2.05 mg / kg, about 2.1 mg / kg, about 2.15 mg / kg, about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg or about 2.5 mg / kg);

[0553] (c) Every three weeks at a dose of about 2.2 mg / kg to about 3.0 mg / kg (e.g., about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg, about 2.5 mg / kg, about 2.55 mg / kg, about 2.6 mg / kg, about 2.65 mg / kg, about 2.7 mg / kg, about 2.75 mg / kg, about 2.8 mg / kg, about 2.85 mg / kg, about 2.9 mg / kg, about 2.95 mg / kg, or about 3.0 mg / kg); or

[0554] (d) Administer at a dose of about 2.5 mg / kg to about 4.2 mg / kg (e.g., about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4.0 mg / kg, about 4.1 mg / kg or about 4.2 mg / kg) once every four weeks.

[0555] In some embodiments, the method further includes reducing the dose range of the antibody or its antigen-binding fragment.

[0556] definition

[0557] As used herein, the term “about” means a value that is more than 10% higher or lower than the stated value. For example, the term “about 5 nM” indicates a range of 4.5 nM to 5.5 nM.

[0558] As used herein, the term "antibody" (Ab) refers to an immunoglobulin molecule that specifically binds to or reacts with a specific antigen, and includes polyclonal, monoclonal, genetically engineered, and other modified forms of antibodies, including but not limited to chimeric antibodies, humanized antibodies, primate-derived antibodies, heteroconjugated antibodies (e.g., bispecific, trispecific, and tetraspecific antibodies, bifunctional, trifunctional, and tetrafunctional antibodies), and antigen-binding fragments of antibodies, including, for example, Fab', F(ab')2, Fab, Fv, IgG, and scFv fragments. Furthermore, unless otherwise indicated, the term "monoclonal antibody" (mAb) is intended to include both the complete molecule and antibody fragments capable of specifically binding to a target protein (such as, for example, Fab and F(ab')2 fragments).

[0559] As used herein, the term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to a target antigen. The antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Antibody fragments can be Fab, F(ab')2, scFv, SMIP, bifunctional antibodies, trifunctional antibodies, affinity antibodies, nanobodies, aptamers, or domain antibodies. Examples of binding fragments covered by the term "antigen-binding fragment" of an antibody include, but are not limited to: (i) Fab fragments, i.e., fragments composed of V... L Structural domain, V H Domain, C L Domain and C H (ii) a monovalent segment composed of a structural domain; (iii) a divalent segment comprising two Fab segments connected by disulfide bonds at the hinge region; and (iv) an Fd segment composed of V H Domain and C H 1. Composition of structural domains; (iv) Fv fragment, said Fv fragment being composed of the V of the single arm of the antibody. L and V H The structural domain is composed of (v)dAb, which includes V H Domain and V L Structural domain; (vi) dAb fragment (Ward et al., Nature 341:544-546, 1989), the dAb fragment being composed of V H Structural domain composition; (vii) dAb, wherein dAb is composed of V H Domain or V LThe structural domains consist of: (viii) separate complementarity-determining regions (CDRs); and (ix) combinations of two or more separate CDRs, which may optionally be connected by a synthesis joint. Furthermore, although the two structural domains V of the Fv fragment... L and V H Encoded by separate genes, but the two domains can be combined using recombination methods via linkers that enable the two domains to become a single protein chain, in which V L District and V H Regions pair to form monovalent molecules (referred to as single-chain Fvs (scFvs); see, for example, Bird et al., Science 242:423-426, 1988, and Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883, 1988). These antibody fragments can be obtained using conventional techniques known to those skilled in the art, and said fragments can be screened for utility in the same manner as intact antibodies. "Antigen-binding fragment" can also refer to linear or nonlinear peptides, which can be cyclic, bicyclic, and / or conformationally biased. Those skilled in the art will understand that conformationally biased antigen-binding fragments will require structure-based design such that the peptide comprises an antigen-binding site mimic based on the 3D structure of the peptide-antigen complex. Such structural information enables the design and generation of continuous and sequentially discontinuous antigen-binding site mimics, which consist of two or more protein segments that are geographically distant in the protein sequence but spatially close through protein folding. Mimicking such discontinuous antigen-binding sites through synthetic peptides typically involves splicing and / or molecular scaffolds (e.g., disulfide bonds) to achieve conformational bias. See, for example, Groß et al., Front. Bioeng. Biotechnol. 4(39), 2016. Antigen-binding fragments can be generated by recombinant DNA technology, enzymatic or chemical cleavage of intact immunoglobulins, or, in some embodiments, by chemical peptide synthesis procedures known in the art.

[0560] As used herein, the terms “agonistic TNFRSF antibody,” “agonistic anti-TNFRSF antibody,” and “TNFRSF agonist antibody” refer to antibodies capable of inducing or promoting the activation of TNFRSF member proteins (e.g., TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, or DCR3), enhancing one or more signal transduction pathways mediated by TNFRSF member proteins, and / or inducing or promoting at least one active TNFRSF antibody mediated by the activation of TNFRSF member proteins. For example, agonistic TNFRSF antibodies can induce or promote the growth and proliferation of T-reg cells. Agonisttic TNFRSF antibodies can induce or promote TNFRSF activation by allosteric binding to TNFRSF. In this way, agonist TNFRSF antibodies can promote or stabilize the trimerization of TNFRSF member proteins, which is achieved through binding to one or more ligands in the TNF superfamily (e.g., CD40L of CD40, TNFα of TNFR1 and TNFR2, 4-1BBL of 4-1BB, CD70 of CD27, CD153 of CD30, N-APP of DR6, EDA-A1 of EDAR, FasL of Fas, GITRL of GITR, LTα of HVEM, LT- The activity of TNFRSF member proteins is induced by the interaction of the β-receptor complex with LT-β (TNF-C), NGF of NGFR, TRAIL of OPG, OX40L of OX40, RANKL of RANK, TRAIL-R2 (TNFRSF10B), TRAIL-R1 (TNFRSF10A), TRAIL of TRAIL-R4, TL1A of TRAMP, EDA-A2 of XEDAR, or FasL, LIGHT, and TL1A of DCR3, etc.

[0561] As used herein, the terms "antagonistic TNFRSF antibody," "antagonistic anti-TNFRSF antibody," and "TNFRSF antagonist antibody" refer to antibodies that can inhibit or reduce the activation of TNFRSF member proteins (e.g., TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, or DCR3), attenuate one or more signal transduction pathways mediated by TNFRSF member proteins, and / or reduce or inhibit at least one active TNFRSF antibody mediated by the activation of TNFRSF member proteins. For example, antagonistic TNFRSF antibodies can inhibit or reduce the growth and proliferation of T-reg cells. Antagonistic TNFRSF antibodies can inhibit or reduce TNFRSF activation by blocking the binding of TNFRSF to one or more ligands in the TNF superfamily (e.g., CD40L of CD40, TNFα of TNFR1 and TNFR2, 4-1BBL of 4-1BB, CD70 of CD27, CD153 of CD30, N-APP of DR6, EDA-A1 of EDAR, FasL of Fas, GITRL of GITR, LTα of HVEM, LT-β (TNF-C) of the LT-β receptor complex, NGF of NGFR, TRAIL of OPG, OX40L of OX40, RANKL of RANK, TRAIL-R2 (TNFRSF10B), TRAIL-R1 (TNFRSF10A), TRAIL-R4 of TRAMP, TL1A, EDA-A2 of XEDAR, or FasL, LIGHT, and TL1A of DCR3). In this way, antagonistic TNFRSF antibodies can block the trimerization of TNFRSF member proteins (which would otherwise be induced by interaction with one or more ligands in the TNF superfamily, such as CD40L, TNFα, 4-1BBL, CD70, CD153, or RANKL), thereby inhibiting the activity of TNFRSF member proteins.

[0562] As used herein, the term "bispecific antibody" refers to an antibody (e.g., a monoclonal antibody, typically a human antibody or a humanized antibody) that has binding specificity against at least two different antigens. For example, one of the binding specificities may target a TNFRSF member protein (e.g., TNFR2, TNFR1, CD40, 4-1BB, CD27, CD30, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R2 (TNFRSF10B), TRAIL-R1 (TNFRSF10A), TRAIL-R4, TRAMP, TROY, XEDAR, or DCR3), and the other binding specificities may target any other antigen, such as a cell surface protein, receptor, receptor subunit, tissue-specific antigen, viral protein, virus-encoded envelope protein, bacterial protein, or bacterial surface protein.

[0563] As used herein, the term "chimeric antibody" refers to an antibody having a variable domain sequence (e.g., a CDR sequence) of an immunoglobulin derived from one source organism (such as a rat or mouse) and a constant region of an immunoglobulin derived from a different organism (e.g., human, non-human primates, pig, goat, rabbit, hamster, cat, dog, guinea pig, various Bovidae family (such as domestic cattle, cattle, bison, buffalo, elk, and yak), sheep, or horse). Methods for generating chimeric antibodies are known in the art. See, for example, Morrison, Science 229:1202-7, 1985; Oi et al., BioTechniques 4:214-221, 1986; Gillies et al., J. Immunol. Methods 125:191-202, 1985; U.S. Patent Nos. 5,807,715, 4,816,567, and 4,816,397, which are incorporated herein by reference.

[0564] As used herein, the term "bivalent antibody" refers to a bivalent antibody comprising two polypeptide chains, each polypeptide chain including V antibodies linked by a linker. H Domain and V LThe linker is too short (e.g., a linker consisting of five amino acids) to allow intramolecular association of the VH and VL domains on the same peptide chain. This configuration forces each domain to pair with a complementary domain on another peptide chain to form a homodimeric structure. Therefore, the term "trifunctional antibody" refers to a trivalent antibody comprising three peptide chains, each containing a VH domain and a VL domain linked by a linker that is too short (e.g., a linker consisting of 1-2 amino acids) to allow intramolecular association of the VH and VL domains on the same peptide chain. To fold into its native structure, peptides configured in this way typically trimerize to spatially close the VH and VL domains of adjacent peptide chains, thereby allowing proper folding (see Holliger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448, 1993; incorporated herein by reference).

[0565] As used herein, the term "monoclonal antibody" refers to an antibody derived from a single clone (including any eukaryotic, prokaryotic, or phage clone) and not to a method of producing the antibody.

[0566] As used herein, the term "multispecific antibody" refers to an antibody that exhibits affinity for more than one target antigen. Multispecific antibodies can have a structure similar to that of a complete immunoglobulin molecule and include an Fc region, such as the IgG Fc region. Such structures can include, but are not limited to, IgG-Fv, IgG-(scFv)2, DVD-Ig, (scFv)2-(scFv)2-Fc, and (scFv)2-Fc-(scFv)2. In the case of IgG-(scFv)2, the scFv can attach to the N-terminus or C-terminus of either the heavy or light chain. Exemplary multispecific molecules including an Fc region and potentially incorporating anti-TNFRSF antibodies or their antigen-binding fragments have been reviewed in the following literature: Kontermann, mAbs 4:182-197, 2012; Yazaki et al., ProteinEngineering, Design & Selection 26:187-193, 2013; and Grote et al., 2012, Proetzel & Ebersbach (eds.), Antibody Methods and Protocols, Methods in Molecular Biology Vol. 901, Chapter 16:247-263; which are incorporated herein by reference. In some embodiments, based on fragments of IgG, DVD, or scFv, the antibody fragment may be a component of a multispecific molecule without an Fc region. Exemplary multispecific molecules lacking an Fc region and capable of incorporating antibodies or antibody fragments, including scFv dimers (bifunctional antibodies), trimers (trifunctional antibodies), and tetramers (tetrafunctional antibodies), Fab dimers (conjugated via adherent peptide or protein domains), and Fab trimers (chemically conjugated), are described by Hudson and Souriau, Nature Medicine 9:129-134, 2003; incorporated herein by reference.

[0567] As used herein, the term "human antibody" refers to virtually every part of a protein (e.g., CDR, framework, C...). L Domain, C H Structural domains (e.g., C) H 1. C H 2. C H 3) Hinges, V L and V HHuman antibodies are essentially non-immunogenic in humans, exhibiting only minor sequence changes or variations. Human antibodies can be produced in human cells (e.g., through recombinant expression) or by non-human animal or prokaryotic or eukaryotic cells capable of expressing functionally rearranged human immunoglobulin (e.g., heavy and / or light chain) genes. Additionally, when the human antibody is a single-chain antibody, it may include a adaptor peptide not present in naturally occurring human antibodies. For example, Fv may contain adaptor peptides linked to variable regions of the heavy chain and light chain, such as two to approximately eight glycine or other amino acid residues. Such adaptor peptides are considered to be of human origin. Human antibodies can be prepared using a variety of methods known in the art, including phage display methods using antibody libraries derived from human immunoglobulin sequences. See U.S. Patents 4,444,887 and 4,716,111; and PCT Publications WO 1998 / 46645; WO 1998 / 50433; WO 1998 / 24893; WO 1998 / 16654; WO 1996 / 34096; WO 1996 / 33735; and WO 1991 / 10741; which are incorporated herein by reference. Human antibodies can also be produced using transgenic mice that do not express functional endogenous immunoglobulins but do express human immunoglobulin genes. See, for example, PCT Publications WO 98 / 24893; WO 92 / 01047; WO 96 / 34096; WO 96 / 33735; U.S. Patent Nos. 5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016; 5,545,806; 5,814,318; 5,885,793; 5,916,771; and 5,939,598, which are incorporated herein by reference.

[0568] As used herein, the term "humanized" antibody refers to a form of non-human (e.g., murine) antibody that is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (such as Fv, Fab, Fab', F(ab')2, or other target-binding subdomains of the antibody) containing a minimal sequence derived from a non-human immunoglobulin. Typically, a humanized antibody will contain at least one of substantially all, and usually two, variable domains, wherein all or substantially all of the CDR regions correspond to the CDR regions of the non-human immunoglobulin. All or substantially all of the FR regions may also be FR regions of a human immunoglobulin sequence. Humanized antibodies may also contain at least a portion of the immunoglobulin constant region (Fc), typically the Fc of a common sequence of human immunoglobulins. Methods for antibody humanization are known in the art. See, for example, Riechmann et al., Nature 332:323-7, 1988; U.S. Patent Nos. 5,225,539, 5,530,101, 5,585,089, 5,693,761, 5,693,762, and 6,180,370; EP239400; PCT Publication WO 91 / 09967; EP592106; and EP519596, which are incorporated herein by reference.

[0569] As used herein, the term "primate-derived antibody" refers to an antibody comprising a framework region derived from a primate-derived antibody and other regions (such as CDRs and / or constant regions) derived from a non-primate-derived antibody. Methods for generating primate-derived antibodies are known in the art. See, for example, U.S. Patent Nos. 5,658,570, 5,681,722, and 5,693,780, which are incorporated herein by reference. For example, the primate-derived antibody or antigen-binding fragment thereof described herein can be generated by inserting a CDR of a non-primate antibody or antigen-binding fragment thereof into an antibody or antigen-binding fragment containing one or more framework regions of a primate.

[0570] As used herein, the term "scFv" refers to a single-chain Fv antibody in which the variable domains of the antibody's heavy and light chains are linked to form a single chain. An scFv fragment contains a single polypeptide chain comprising variable regions (e.g., CDR-L1, CDR-L2, and / or CDR-L3) of the antibody light chain (VL) and variable regions (e.g., CDR-H1, CDR-H2, and / or CDR-H3) of the antibody heavy chain (VH), separated by linkers. The linkers connecting the VL and VH regions of the scFv fragment can be peptide linkers composed of proteogenic amino acids. Alternative linkers can be used to enhance the resistance of scFv fragments to proteolytic degradation (e.g., linkers containing D amino acids), improve the solubility of scFv fragments (e.g., hydrophilic linkers, such as linkers containing polyethylene glycol or peptides containing repeating glycine and serine residues), improve the biophysical stability of molecules (e.g., linkers containing cysteine ​​residues that form intramolecular or intermolecular disulfide bonds), or reduce the immunogenicity of scFv fragments (e.g., linkers containing glycosylation sites). scFv molecules are known in the art and have been described, for example, in U.S. Patent No. 5,892,019; Flo et al., Gene77:51, 1989; Bird et al., Science 242:423, 1988; Pantolian et al., Biochemistry 30:10117, 1991; Milenic et al., Cancer Research 51:6363, 1991; and Takkinen et al., Protein Engineering 4:837, 1991. The VL and VH domains of scFv molecules can be derived from one or more antibody molecules. Those skilled in the art will also understand that the variable regions of the scFv molecules described herein can be modified so that their amino acid sequences differ from those of the antibody molecules from which they are derived. For example, in some embodiments, nucleotide or amino acid substitutions can be made, resulting in conserved substitutions or changes at amino acid residues (e.g., in CDR and / or framework residues). Alternatively or additionally, CDR amino acid residues can be mutated using techniques recognized in the art to optimize antigen binding. scFv fragments are described, for example, in WO 2011 / 084714; which is incorporated herein by reference.

[0571] As used herein, a “disulfide-bonded isoform” of an antibody or its antigen-binding fragment is a form of antibody or its antigen-binding fragment having a specific internal disulfide bonding pattern. A disulfide-bonded isoform is a structural isoform of a given antibody or its antigen-binding fragment that is identical to each other in amino acid sequence but exhibits different disulfide bond connections. For example, in the context of human IgG2 antibodies or variants thereof, the antibody may be present in one of four possible disulfide-bonded isoforms (referred herein to as isoforms IgG2-A, IgG2-B, IgG2-A / B1, and IgG2-A / B2).

[0572] As used herein, the term “complementarity-determining region” (CDR) refers to a hypervariable region found in both the light chain and heavy chain variable domains of an antibody. The more conserved portion of the variable domain is called a frame region (FR). As understood in the art, the amino acid positions describing the hypervariable regions of an antibody can vary depending on the context and various definitions known in the art. Some positions within a variable domain can be considered hybrid hypervariable positions because they can be considered within a hypervariable region under one set of criteria and outside a hypervariable region under a different set of criteria. One or more of these positions can also be found in extended hypervariable regions. The antibodies described herein may contain modifications in these hybrid hypervariable positions. The variable domains of the native heavy and light chains each contain four frame regions that primarily employ a β-sheet configuration linked by three CDRs, which form loops connected to the β-sheet structure and, in some cases, form part of the β-sheet structure. The CDRs in each chain are held together by the FR regions in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, and together with the CDRs from other antibody chains, they contribute to the formation of the antibody's target binding site (see Kabat et al., Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md. 1987); incorporated herein by reference). As used herein, unless otherwise stated, the numbering of immunoglobulin amino acid residues is based on the EU numbering system (see, for example, Edelman et al., Proc. Natl. Acad. Sci. USA 63:78-85, 1969; incorporated herein by reference). CDRs can be identified using sequence- or structure-based methods already described by Kabat et al., Chothia et al. (J. Mol. Biol. 196:901-917, 1987), and MacCallum et al. (J. Mol. Biol. 262:732-745, 1996), where the definition includes overlap or subsets of amino acid residues when compared with each other. The term "CDR" can be, for example, a CDR defined by Kabat or Chothia based on sequence comparison.

[0573] As used herein, the term "frame region" or "FW region" includes the amino acid residues adjacent to the CDR. FW region residues may be present in, for example, human antibodies, rodent-derived antibodies (e.g., mouse antibodies), humanized antibodies, primate-derived antibodies, chimeric antibodies, antibody fragments (e.g., Fab fragments), single-chain antibody fragments (e.g., scFv fragments), antibody domains, and bispecific antibodies.

[0574] As used herein, the terms “conservative mutation,” “conservative substitution,” or “conservative amino acid substitution” refer to the substitution of one or more different amino acids that exhibit similar physicochemical properties (such as polarity, electrostatic charge, and / or steric volume). Table 1 below summarizes these properties for each of the twenty naturally occurring amino acids.

[0575] Table 1. Representative physicochemical properties of naturally occurring amino acids.

[0576]

[0577] As can be understood from this table, conserved amino acid families include, but are not limited to: (i) G, A, V, L, I, P, and M; (ii) D and E; (iii) C, S, and T; (iv) H, K, and R; (v) N and Q; and (vi) F, Y, and W. Therefore, a conserved mutation or substitution refers to replacing an amino acid with a member of the same amino acid family (e.g., replacing Thr with Ser, or Arg with Lys). As understood in the art, peptides can retain their biological properties or functions upon the introduction of one or more conserved substitutions.

[0578] Amino acid substitutions can be represented herein by the conventions (AA1)(N)(AA2), where “AA1” indicates an amino acid normally present at a specific site within the amino acid sequence, “N” indicates the number of residues in the amino acid sequence in which the substitution occurs, and “AA2” indicates the amino acid present in the amino acid sequence after the substitution is achieved. For example, in the context of an antibody hinge region (such as the IgG2 antibody hinge region), the symbol “C232S” indicates that a serine residue at amino acid residue 232 of the indicated hinge amino acid sequence has been substituted with a naturally occurring cysteine ​​residue. Similarly, in the context of an antibody hinge region (such as the IgG2 antibody hinge region), the symbol “C233S” indicates that a serine residue at amino acid residue 233 of the indicated hinge amino acid sequence has been substituted with a naturally occurring cysteine ​​residue.

[0579] As used herein, the term "hydrophobic side chain" refers to an amino acid side chain that exhibits relatively low solubility in water due to the spatial or electronic properties of the chemical moiety present within the side chain. Examples of amino acids containing hydrophobic side chains include amino acids containing aliphatic hydrocarbons, such as alanine, valine, leucine, isoleucine, proline, and methionine, as well as amino acids containing electrostatically neutral aromatic ring systems at physiological pH, such as tryptophan, phenylalanine, and tyrosine.

[0580] As used herein, the term "conjugate" refers to a compound formed by the chemical bonding of a reactive functional group of one molecule with a suitable reactive functional group of another molecule.

[0581] As used herein, the term "chemotherapeutic agent" refers to any chemical agent that is therapeutically useful in the treatment of cancers, such as those described herein. Chemotherapy agents encompass both chemical and biological agents. These agents work by inhibiting the cellular activity upon which cancer cells depend for continued survival. Categories of chemotherapeutic agents include alkylating agents / alkaloids, antimetabolites, hormones, hormone analogs, and antitumor drugs. Exemplary chemotherapeutic agents suitable for use in conjunction with the compositions and methods described herein include, but are not limited to, those described in the following literature: Slapak and Kufe, Principles of Cancer Therapy, Chapter 86 Harrison's Principles of Internal Medicine, 14th ed.; Perry et al., Chemotherapeutic, Chapter 17 Abeloff, Clinical Oncology 2nd ed., 2000; Baltzer L. and Berkery R. (eds.): Oncology Pocket Guide to Chemotherapeutic, 2nd ed. St. Louis, Mosby-Year Book, 1995; Fischer DS, Knobf MF, Durivage HJ (eds.): The Cancer Chemotherapeutic Handbook, 4th ed. St. Louis, Mosby-Year Handbook. Since they relate to chemotherapeutic agents, the disclosure of each of these literatures is incorporated herein by reference.

[0582] As used herein, the term "immunotherapy agent" refers to a compound, such as an antibody, its antigen-binding fragment, single-chain polypeptide, or construct as described herein, that specifically binds to and antagonizes an immune checkpoint protein (e.g., an immune checkpoint receptor or ligand), thereby reducing or inhibiting receptor or ligand signal transduction that would otherwise lead to a downregulation of the immune response. Immunotherapy agents include compounds, such as antibodies, antigen-binding fragments, single-chain polypeptides, and constructs, that are capable of specifically binding to receptors expressed on the surface of hematopoietic cells (e.g., lymphocytes, T cells) and inhibiting receptor or ligand-induced signal transduction that would otherwise lead to tolerance to endogenous ("self") antigens (e.g., tumor-associated antigens). Compared to receptor- or ligand-induced signaling observed in the absence of an immunotherapy agent, an immunotherapy agent can reduce receptor- or ligand-induced signaling by, for example, 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100%. Exemplary assays that can be used to measure the extent of receptor or ligand signaling include, for example, enzyme-linked immunosorbent assay (ELISA) techniques for measuring changes in protein expression associated with a specific signal transduction pathway, and polymerase chain reaction (PCR)-based techniques, such as quantitative PCR, reverse transcription PCR, and real-time PCR experiments, which can be used to determine changes in gene expression associated with a specific signal transduction pathway. Exemplary methods that can be used to determine whether an agent is an “immunotherapy agent” include the assays described in Mahoney et al., Cancer Immunotherapy 14:561-584, 2015, the disclosure of which is incorporated herein by reference in its entirety. Examples of immunotherapy agents include, for example, antibodies or antigen-binding fragments thereof that specifically bind to one or more of the following: TL1A, CD40L, LIGHT, BTLA, LAG3, TIM3, Singlecs, ICOS, B7-H3, B7-H4, VISTA, TMIGD2, BTNL2, CD48, KIR, LIR, LIR antibody, ILT, NKG2D, NKG2A, MICA, MICB, CD244, CSF1R, IDO, TGFβ, CD39, CD73, CXCR4, CXCL12, SIRPA, CD47, VEGF, and neurofeltin. Further examples of immunotherapy agents include targretin, interferon alpha, clobetasol, and Peg interferon (e.g., PEGASYS). ®Prednisone, Romidepsin, Bexarotene, Methotrexate, Triamcinolone cream, antichemokines, Vorinostat, Gabapentin, antibodies against lymphocyte surface receptors and / or lymphokines, antibodies against surface cancer proteins, and / or small molecule therapies such as Vorinostat. Specific examples of immunotherapeutic agents that may be used in or in combination with the compositions and methods described herein include anti-PD-1 antibodies and their antigen-binding fragments such as nivolumab, pembrolizumab, avelumab, durvalumab, and atezolizumab; anti-PD-L1 antibodies and their antigen-binding fragments such as atezolizumab and avelumab; and anti-CTLA-4 antibodies and their antigen-binding fragments such as ipilimumab or tremelimumab.

[0583] As used herein in the context of TNFRSF agonists or antagonists, the term "construction" refers to a fusion protein containing a first polypeptide domain that binds to a second polypeptide domain. The polypeptide domains can each be independently agonist or antagonist antigen-binding fragments (e.g., single-chain polypeptides as described herein) that bind to a TNFRSF member protein. The first polypeptide domain can be covalently bound to the second polypeptide domain, for example, through a linker (such as a peptide linker or disulfide bond). Exemplary linkers that can be used to link polypeptide domains to TNFRSF agonist or antagonist constructs include, but are not limited to, the linker described in Leriche et al., Bioorg. Med. Chem. 20:571-582, 2012, the disclosure of which is incorporated herein by reference in its entirety.

[0584] As used herein, the term “endogenous” describes molecules (e.g., polypeptides, nucleic acids, or cofactors) that are naturally present in a particular organism (e.g., humans) or at a specific location within an organism (e.g., an organ, tissue, or cell, such as a human cell).

[0585] As used herein, the term "exogenous" describes molecules (e.g., polypeptides, nucleic acids, or cofactors) that are not naturally present in a particular organism (e.g., human) or at a specific location within an organism (e.g., an organ, tissue, or cell, such as a human cell). Exogenous materials include materials that are provided to or cultured from an external source.

[0586] As used herein, the term "epitope" refers to a portion of an antigen that is recognized and bound by a polypeptide (such as an antibody, its antigen-binding fragment, a single-chain polypeptide, or a construct described herein). In the context of protein antigens (such as human TNFRSF member proteins), an epitope can be a continuous epitope, which is a single, uninterrupted segment of one or more amino acids covalently linked together by peptide bonds, wherein all component amino acids bind to the polypeptide (e.g., an antibody, its antigen-binding fragment, a single-chain polypeptide, or a construct thereof). A continuous epitope can consist of, for example, 1, 5, 10, 15, 20, or more amino acids within an antigen (such as human TNFRSF member proteins). In some embodiments, an epitope can be a discontinuous epitope, which comprises two or more amino acids, each separated from each other by one or more intermediate amino acid residues in the amino acid sequence of the antigen. Despite this separation by intermediate amino acids, the segments constituting the discontinuous epitope may, for example, be spatially close to each other in the three-dimensional conformation of the antigen.

[0587] As used herein, the term "fusion protein" refers to a protein covalently linked to another molecule. Fusion proteins can be chemically synthesized, for example, by an amide bond formation reaction between the N-terminus of one protein and the C-terminus of another protein. Alternatively, fusion proteins containing a protein covalently bound to another protein can be recombinantly expressed in cells (e.g., eukaryotic or prokaryotic cells) by expressing a polynucleotide encoding the fusion protein (e.g., from the genome of a vector or cell). Fusion proteins may contain a protein covalently bound to a linker, which in turn covalently binds to another molecule. Examples of linkers that can be used to form fusion proteins include peptide linkers, such as linkers containing naturally occurring or non-naturally occurring amino acids. In some embodiments, it may be necessary to include D-amino acids in the linker because these residues are not present in naturally occurring proteins and are therefore more resistant to degradation by endogenous proteases. The linker can be prepared using a variety of strategies well known in the art, and depending on the reactive components of the linker, it can be cleaved by enzymatic hydrolysis, photolysis, hydrolysis under acidic conditions, degradation under alkaline conditions, oxidation, disulfide reduction, nucleophilic cleavage, or organometallic cleavage (Leriche et al., Bioorg. Med. Chem. 20:571-582, 2012).

[0588] As used herein, the term "hinge region" refers to a domain of an antibody or antigen-binding fragment (e.g., an IgG2 antibody or its antigen-binding fragment) located between the antigen-binding portion of the antibody or its antigen-binding fragment (such as the Fab region of the antibody or its antigen-binding fragment) and the portion of the antibody or its antigen-binding fragment that determines the type of said antibody or its antigen-binding fragment (such as the Fc region of the antibody or its antigen-binding fragment). For example, in the context of monoclonal antibodies, the hinge region is a polypeptide located approximately at the center of each heavy chain, thereby connecting the CH1 domain with the CH2 and CH3 domains. The hinge region of an antibody or its antigen-binding fragment can provide chemical bonds between the chains of the antibody or its antigen-binding fragment. For example, in monoclonal antibodies, cysteine ​​residues within the hinge region form interchain disulfide bonds, thereby providing well-defined covalent bonds between the heavy chains. The amino acid sequence of wild-type human IgG2 is ERKCCVECPPCP (SEQ ID NO: 51). As used herein, the antibody hinge region is numbered according to the numbering system of Kabat et al., Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md. 1987), the contents of which are incorporated herein by reference.

[0589] As used herein, the term "myeloid-derived suppressor cells" or "MDSCs" refers to cells of the immune system that regulate the activity of various effector cells and antigen-presenting cells, such as T cells, NK cells, dendritic cells, and macrophages. Myeloid-derived suppressor cells are distinguished by their gene expression profiles, expressing all or a subset of proteins and small molecules selected from the following groups: B7-1 (CD80), B7-H1 (PD-L1), CCR2, CD1d, CD1d1, CD2, CD31 (PECAM-1), CD43, CD44, complement component C5a R1, F4 / 80 (EMR1), Fcγ RIII (CD16), Fcγ RII (CD32), Fcγ RIIA (CD32a), Fcγ RIIB (CD32b), Fcγ RIIB / C (CD32b / c), Fcγ RIIC (CD32c), Fcγ RIIIA (CD16A), Fcγ RIIIB (CD16b), galactagogue-3, GP130, Gr-1 (Ly-6G), ICAM-1 (CD54), IL-1RI, IL-4Rα, IL-6Rα, integrin α4 (CD49d), integrin αL (CD11a), integrin αM (CD11b), M-CSFR, MGL1 (CD301a), MGL1 / 2 (CD301a / b), MGL2 (CD301b), nitric oxide, PSGL-1 (CD162), L-selectin (CD62L), siglec-3 (CD33), transferrin receptor (TfR), VEGFR1 (Flt-1), and VEGFR2 (KDR or Flk-1). Specifically, MDSCs do not express proteins selected from the following groups: B7-2 (CD86), B7-H4, CD11c, CD14, CD21, CD23 (FcεRII), CD34, CD35, CD40 (TNFRSF5), CD117 (c-kit), HLA-DR, and Sca-1 (Ly6).

[0590] As used herein, the term "percentage of sequence identity (%)" refers to the percentage of amino acid (or nucleic acid) residues in a candidate sequence that are identical to those in a reference sequence after alignment and the introduction of gaps (if necessary) to achieve the maximum percentage of sequence identity (e.g., gaps may be introduced in one or both of the candidate and reference sequences for optimal alignment, and non-homologous sequences may be ignored for comparison purposes). Alignments used to determine the percentage of sequence identity can be performed in various ways within the scope of the art, such as using publicly available computer software like BLAST, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignments, including any algorithm required to achieve maximum alignment across the full length of the sequences being compared. For example, a reference sequence aligned to be compared with a candidate sequence may indicate that the candidate sequence exhibits 50% to 100% sequence identity across the full length of the candidate sequence or selected portions of consecutive amino acid (or nucleic acid) residues of the candidate sequence. The length of candidate sequences compared for comparative purposes can be, for example, at least 30% of the length of the reference sequence (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%). Molecules are considered identical at said position when a position in the candidate sequence is occupied by the same amino acid residue as the corresponding position in the reference sequence.

[0591] As used herein, the term “proliferation” in the context of a population of cells (such as a population of cells expressing TNFRSF, e.g., T-reg cells, MDSCs, or cancer cells expressing TNFRSF) refers to the mitotic and kinetic division of cells to produce multiple cells. Cell proliferation can be demonstrated, for example, by finding that the number of cells (e.g., cells expressing TNFRSF) in a sample of a subject or cells has increased over a given period of time (e.g., over an hour or more, a day or more, or a week or more). Those skilled in the art can monitor cell proliferation using a variety of known techniques (e.g., by visual microscopy, hemocytology, flow cytometry, fluorescence-activated cell sorting, and other assays known in the art). In this disclosure, a change (e.g., an increase or decrease) in cell proliferation is considered to have occurred when the proliferation rate of a population of cells (such as a population of TNFRSF-expressing cells in contact with an agonist or antagonist TNFRSF antibody or its antigen-binding fragment as described herein) changes relative to the proliferation rate of a population of control cells (such as a population of TNFRSF-expressing cells not in contact with an agonist or antagonist TNFRSF antibody or its antigen-binding fragment). Changes in proliferation rate can manifest as, for example, changes in the amount of cells of interest in a subject or sample over a given period of time (e.g., an increase or decrease), such as changes in the amount of cells of interest in a subject or sample over a given period of time of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or more. Alternatively, changes in cell proliferation can be demonstrated by the following finding: a change (e.g., an increase or decrease) in the rate of division of the cells of interest (e.g., TNFRSF-expressing cells in contact with the agonist or antagonist TNFRSF antibody or its antigen-binding fragment as described herein), for example, relative to the rate of division of control cells (e.g., TNFRSF-expressing cells not in contact with the agonist or antagonist TNFRSF antibody or its antigen-binding fragment), by 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or more.

[0592] As used herein, the term "pharmacokinetic profile" refers to the absorption, distribution, metabolism, and clearance of a drug over time after administration to a subject.

[0593] As used herein, the terms “natural TNFRSF ligand,” “endogenous TNFRSF ligand,” and “endogenous TNFRSF ligand” refer to endogenous ligands of TNFRSF member proteins that can form TNFRSF ligand-TNFRSF member protein complexes and induce the formation of an active homotrimeric conformation of TNFRSF member proteins. Exemplary TNFRSF ligand-TNFRSF member protein complexes may include, but are not limited to, CD40L-CD40 complex, 4-1BBL-4-1BB complex, CD70-CD27 complex, CD153-CD30 complex, N-APP-DR6 complex, EDA-A1-EDAR complex, FasL-Fas complex, GITRL-GITR complex, LT-α-HVEM complex, LT-β(TNF-C)-LT-β receptor complex, NGF-NGFR complex, TRAIL-OPG complex, OX40L-OX40 complex, RANKL-RANK complex, TNFα-TNFR1 complex, TRAIL-TRAIL-R2 (TNFRSF10B) complex, TRAIL-TRAIL-R1 (TNFRSF10A) complex, TRAIL-TRAIL-R4 complex, TL1A-TRAMP complex, EDA-A2-XEDAR complex, FasL-DCR3 complex, LIGHT-DCR3 complex, and TL1A-DCR3 complex.

[0594] As used herein, the phrase "specific binding" refers to a binding reaction that determines the presence of an antigen in a heterogeneous population of proteins and other biomolecules, said antigen being specifically recognized by, for example, an antibody or its antigen-binding fragment. Antibodies or their antigen-binding fragments that specifically bind to an antigen will bind at a Kc of less than 100 nM. d Binding to antigens. For example, antibodies or antigen-binding fragments that specifically bind to antigens will have a K+ of up to 100 nM (e.g., between 1 pM and 100 nM). d Binding to antigens. Antibodies or antigen-binding fragments that do not show specific binding to a particular antigen or its epitope will exhibit a K+ value greater than 100 nM (e.g., greater than 500 nm, 1 µM, 100 µM, 500 µM, or 1 mM) against that specific antigen or epitope. dVarious immunoassays can be used to select antibodies that have a specific immunoreactivity with a particular protein or carbohydrate. For example, solid-phase ELISA is routinely used to select antibodies that have a specific immunoreactivity with a protein or carbohydrate. For a description of the immunoassays and conditions that can be used to determine specific immunoreactivity, see Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Press, New York (1988) and Harlow & Lane, Using Antibodies, A Laboratory Manual, Cold Spring Harbor Press, New York (1999).

[0595] As used herein, the terms “subject” and “patient” refer to an organism receiving treatment for a specific disease or symptom (such as cancer or an infectious disease) as described herein. Examples of subjects and patients include mammals receiving treatment for diseases or symptom (e.g., cell proliferation disorders such as cancer or infectious diseases), such as humans, primates, pigs, goats, rabbits, hamsters, cats, dogs, guinea pigs, members of the Bovidae family (such as domestic cattle, cattle, bison, buffalo, elk, and yaks), sheep, and horses.

[0596] As used herein, the term “transfection” refers to any of the various techniques commonly used to introduce exogenous polynucleotides into prokaryotic or eukaryotic host cells, such as electroporation, lipid transfection, calcium phosphate precipitation, diethylaminoethyl (DEAE)-glucan transfection, etc.

[0597] As used herein, the term "treat" or "treatment" refers to a therapeutic treatment aimed at preventing or slowing (alleviating) the progression of an undesirable physiological change or condition, such as a cell-proliferating condition (e.g., cancer, autoimmune disease, or infectious disease). Beneficial or desired clinical outcomes include, but are not limited to, symptom relief, reduction in disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (whether partial or complete), whether detectable or undetectable (e.g., particularly in subjects not treated with the compositions described herein). Subjects requiring treatment include those who already have the stated condition or symptom, those who are susceptible to the stated condition or symptom, or those who wish to prevent the stated condition or symptom.

[0598] As used herein, the terms “tumor necrosis factor receptor superfamily,” “TNFR superfamily,” “TNFRS,” “TNFRSF,” or “TNFRSF member” refer to a group of type I transmembrane proteins having a carboxyl-terminal intracellular domain and an amino-terminal extracellular domain, characterized by a common cysteine-rich domain (CRD). The TNFR superfamily includes receptors that mediate cell signaling by binding to one or more ligands from the TNF superfamily. The TNFR superfamily can be divided into two subgroups: receptors containing an intracellular death domain and receptors lacking this domain. The death domain is an 80-amino acid motif that propagates the apoptosis signaling cascade upon receptor activation. Exemplary TNFR superfamily members containing an intracellular death domain include TNFR1, while TNFR2 represents a TNFR superfamily protein lacking this domain. Members of the TNFR superfamily include CD40, TNFR1, TNFR2, RANK, CD30, lymphotoxin β receptor (LT-β receptor or LT-βR), OX40, Fas receptor, decoy receptor 3 (DCR3), CD27, 4-1BB, death receptor 4 (DR4), death receptor 5 (DR5), decoy receptor 1 (DCR1), decoy receptor 2 (DCR2), osteoprotein, TWEAK receptor, TACI, BAFF receptor, herpesvirus entry mediator, nerve growth factor receptor, B cell maturation antigen, glucocorticoid-induced TNFR-related protein, TROY, death receptor 6 (DR6), death receptor 3 (DR3), and exoprotein A2 receptor.

[0599] As used herein, the terms “TNFRSF signaling,” “TNFRSF member protein signaling,” and “TNFRSF signal transduction” are used interchangeably and refer to TNFRSF members (e.g., TNFR2, TNFR1, CD40, 4-1BB, CD27, CD30, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R2 (TNFRSF10B), TRAIL-R1 (TNFRSF10A), TRAIL-R4, TRAMP, TROY, XEDAR, or D) typically found on the surface of cells expressing TNFRSF members (such as T-reg cells, MDSCs, or cancer cells expressing TNFRSF members). CR3 is a cellular event that typically occurs when it is activated by endogenous TNFRSF member ligands such as CD40L of CD40, TNFα of TNFR1 and TNFR2, 4-1BBL of 4-1BB, CD70 of CD27, CD153 of CD30, N-APP of DR6, EDA-A1 of EDAR, FasL of Fas, GITRL of GITR, LTα of HVEM, LT-β of the LT-β receptor complex (TNF-C), NGF of NGFR, TRAIL of OPG, OX40L of OX40, RANKL of RANK, TRAIL-R2 (TNFRSF10B), TRAIL-R1 (TNFRSF10A), TRAIL of TRAIL-R4, TL1A of TRAMP, EDA-A2 of XEDAR, or FasL, LIGHT, and TL1A of DCR3. TNFRSF member signaling can be demonstrated by the presence of increased expression of one or more genes selected from the group consisting of: CHUK, NFKBIE, NFKBIA, MAP3K11, TRAF2, TRAF3, relB, and cIAP2 / BIRC3. When exposed to an agent (such as an agonist or antagonist TNFRSF antibody or its antigen-binding fragment as described herein), TNFRSF member signaling is considered “inhibited” as used herein when, relative to cells expressing TNFRSF members that have not been exposed to an agent (e.g., an agonist or antagonist TNFRSF antibody or its antigen-binding fragment), the expression (and / or post-translational modifications, if such modifications are required for the activity of the encoded protein) of one or more of the aforementioned genes is reduced.For example, when the expression or post-translational modification (e.g., phosphorylation) of one or more of CHUK, NFKBIE, NFKBIA, MAP3K11, TRAF2, TRAF3, relB, or cIAP2 / BIRC3 in cells expressing TNFRSF members that have been exposed to an antagonistic TNFRSF member peptide is reduced by approximately 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% (relative to the expression or post-translational modification (e.g., phosphorylation) of one or more of these genes in cells expressing TNFRSF members that have not been exposed to an antagonistic TNFRSF member peptide), TNFRSF member signaling is considered “inhibited.” Exemplary assays that can be used to determine expression levels and phosphorylation status are known in the art, including, for example, Western blot assays for determining protein content and quantitative reverse transcription polymerase chain reaction (RT-PCR) experiments for determining mRNA content.

[0600] As used herein, the term "vector" includes nucleic acid vectors, such as DNA vectors, plasmids, RNA vectors, and viruses or other suitable replicons (e.g., viral vectors). A variety of vectors have been developed for the delivery of polynucleotides encoding exogenous proteins into prokaryotic or eukaryotic cells. Examples of such expression vectors are disclosed, for example, in WO 1994 / 11026; incorporated herein by reference. The expression vectors described herein contain polynucleotide sequences and additional sequence elements, for example, for protein expression and / or integration of these polynucleotide sequences into the genome of mammalian cells. Some vectors that can be used for the expression of antibodies and antibody fragments described herein include plasmids containing regulatory sequences, such as promoter and enhancer regions that direct gene transcription. Other useful vectors for the expression of antibodies or antibody fragments contain polynucleotide sequences that enhance the translation rate of these genes or improve the stability or nuclear export of mRNA produced by gene transcription. These sequence elements include, for example, 5' and 3' untranslated regions, internal ribosome entry sites (IRES), and polyadenylation signal sites to guide efficient transcription of the gene carried on the expression vector. The expression vectors described in this article may also contain polynucleotides encoding biomarkers to allow selection of cells containing such vectors. Examples of suitable biomarkers include genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, kanamycin, or nourseothricin.

[0601] As used herein, the term "VH" refers to the variable region of the immunoglobulin heavy chain of an antibody, including the heavy chain of Fv, scFv, or Fab. The term "VL" refers to the variable region of the immunoglobulin light chain, including the light chain of Fv, scFv, dsFv, or Fab. Antibodies (Abs) and immunoglobulins (Igs) are glycoproteins with similar structural properties. While antibodies exhibit binding specificity to a particular target, immunoglobulins include both antibodies and other antibody-like molecules lacking target specificity. Natural antibodies and immunoglobulins are typically heterotetraglycoproteins of about 150,000 Daltons, consisting of two identical light (L) chains and two identical heavy (H) chains. Each heavy chain of a natural antibody has a variable domain (VH) at the amino terminus, followed by numerous constant domains. Each light chain of a natural antibody has a variable domain (VL) at the amino terminus and a constant domain at the carboxyl terminus.

[0602] As used herein, the term "reference level" refers to a threshold level or a level in a control subject or control patient population. The reference level depends on the assay performed and can be determined by those skilled in the art. The reference level can be a baseline level or a level measured in the same subject at an earlier or later time point. In some cases, the reference level in a subject is determined before or after administration of the antibody or antigen-binding fragment thereof disclosed herein, the construct disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the host cell disclosed herein, or the pharmaceutical composition disclosed herein. For example, some non-limiting examples of reference levels for human soluble TNFR2 include human soluble TNFR2 levels in subjects who: have not been diagnosed with a disease; do not have symptoms of at least two or more diseases; or have not been administered the anti-TNFRSF antibody or antigen-binding fragment thereof disclosed herein.

[0603] As used herein, the term "modified antibody or antigen-binding fragment thereof" refers to an antibody or antigen-binding fragment thereof that contains one or more amino acid modifications (e.g., substitution, deletion, insertion, or chemical modification, etc.) (e.g., an antibody or antigen-binding fragment thereof that specifically binds to a TNFRSF member protein, such as the antibody or antigen-binding fragment thereof disclosed herein). For example, an antibody or antigen-binding fragment thereof may have one or more amino acid modifications at the Fc domain. The term "unmodified antibody or antigen-binding fragment thereof" refers to an antibody or antigen-binding fragment thereof that corresponds to a modified antibody or antigen-binding fragment thereof but does not contain one or more amino acid modifications.

[0604] As used herein, the term "biofluid" refers to a source of fluid and includes (but is not limited to) amniotic fluid, aqueous humor, blood and plasma (and unless otherwise expressly indicated by the context, blood refers to plasma components), cerumen (earwax), Cowper's fluid, chyme, interstitial fluid, lymph, mammalian milk, mucus, pleural effusion, pus, saliva, sebum, semen, serum, sweat, urine, vaginal secretions, vomitus, and exudate (from wounds or lesions). In some embodiments, the biofluid of this disclosure is serum. In some embodiments, the biofluid of this disclosure is plasma.

[0605] As used herein, the term "immune response" includes, but is not limited to, T cell-mediated and / or B cell-mediated immune responses. Exemplary immune responses include T cell responses, such as cytokine production and cytotoxicity. T cell responses include Th1 responses and / or Th2 responses. Additionally, the term "immune response" includes responses indirectly promoted by T cell activation (e.g., antibody production (humoral response)) and activation of cytokine-responsive cells (e.g., eosinophils, macrophages). Furthermore, the term "immune response" includes immune responses indirectly influenced by T cell activation (e.g., antibody production (humoral response) and activation of cytokine-responsive cells (e.g., macrophages)). Immune cells involved in the immune response include lymphocytes, such as T cells (CD4+, CD8+, Th1 and Th2 cells, memory T cells) and B cells; antigen-presenting cells (e.g., professional antigen-presenting cells, such as dendritic cells, macrophages, B lymphocytes, Langerhans cells, and non-professional antigen-presenting cells, such as keratinocytes, endothelial cells, astrocytes, fibroblasts, oligodendrocytes); natural killer (NK) cells; and bone marrow cells, such as macrophages, eosinophils, mast cells, basophils, and granulocytes. The immune response can also refer to any of the following: innate immunity, humoral immunity, cellular immunity, autoimmunity, inflammatory response, and acquired (adaptive) immunity.

[0606] As used herein, the term "dosage range" refers to the upper and lower limits of acceptable variation in the amount (e.g., dose) of a specified agent (e.g., an antibody or antigen-binding fragment thereof of this disclosure that specifically binds to a TNFRSF member protein, a TNFSF member protein, CD28, or ICOS). Generally, any amount of the agent within the specified range can be administered to a patient undergoing treatment. Attached Figure Description

[0607] Figure 1A This is a graph showing the viable cell counts of Jeko-1 tumor cells under different concentrations of fully humanized TNFR2 antagonist antibody. The experiment was conducted as described in Example 1 below.

[0608] Figure 1B This is a graph showing the live cell counts of Jeko-1 tumor cells under different concentrations of antibodies against a fully humanized TNFR2 antagonist with reduced interaction with the Fc receptor.

[0609] Figure 2A This is a graph showing the percentage of Treg cells in human T cells treated with a fully humanized TNFR2 agonist antibody. The experiment was conducted as described in Example 2 below.

[0610] Figure 2B This is a graph showing the percentage of Treg cells in human T cells treated with a fully humanized TNFR2 agonist antibody with an LALA mutation in the Fc region.

[0611] Figure 3A This is a graph showing the percentage of Treg cells in human T cells treated with a human anti-human TNFR2 agonist. The experiment was conducted as described in Example 3 below.

[0612] Figure 3B This is a graph showing the percentage of Treg cells in human T cells treated with a human anti-human TNFR2 agonist with LALA mutations in the Fc region.

[0613] Figure 3C This is a graph showing the percentage of Treg cells in human T cells treated with a mouse anti-human TNFR2 agonist.

[0614] Figure 4A This is a graph showing Treg cell proliferation in response to increased concentrations of Ab1 (a modified humanized IgG4 anti-TNFR2 agonist antibody with reduced Fc effector function) (black circle) or the corresponding humanized IgG1 anti-TNFR2 agonist antibody (black square).

[0615] Figure 4B This is a graph showing the change in the percentage of effector T cells (Teff) in total T cells in response to increased concentrations of Ab1 (black circle) or the corresponding humanized IgG1 anti-TNFR2 agonist antibody (black square).

[0616] Figure 5A This is a graph showing the change in the percentage of Treg cells in total T cells in response to an increase in Ab1 concentration.

[0617] Figure 5B This is a graph showing the change in the percentage of effector T cells (Teff) in total T cells in response to an increase in Ab1 concentration.

[0618] Figure 6This is a graph showing the change in the percentage of Treg cells in total T cells in response to increased concentrations of Ab1 or control TNFR2 agonist antibodies with full Fc effector function. Antibody concentrations are shown on a logarithmic scale.

[0619] Figure 7 This diagram illustrates two different modes of complex formation between an agonist antibody and its target cell surface receptor protein. An agonist antibody can form a single complex with a cell surface receptor protein, where the antibody binds to only one cell surface receptor protein. Single complex binding does not activate the cell surface receptor, resulting in low agonist activity. Alternatively, an agonist antibody can form a dual complex with two adjacent cell surface receptor proteins, where the antibody binds to one receptor on each arm. Dual complex binding leads to efficient receptor cross-linking, potent receptor activation, and high agonist activity.

[0620] Figure 8 This shows the change in the agonistic activity of the agonist antibody in response to an increase in antibody concentration.

[0621] Figure 9 This is a graph showing the in vivo concentration of agonistic antibodies after subcutaneous (SQ) or intravenous (IV) administration. SQ administration ensures that the antibody reaches optimal concentrations over time to allow for the formation of dual complexes, thus avoiding the lack of receptor cross-linking that can occur with IV administration due to supersaturation at peak concentrations or insufficient receptor binding at trough concentrations. Detailed Implementation

[0622] The antibody or antigen-binding fragment thereof of this disclosure that specifically binds to human TNFRSF member proteins (e.g., TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, or DCR3, etc.), TNFSF member proteins (e.g., TRAIL), CD28, or ICOS contains one or more amino acid modifications (e.g., substitution, deletion, insertion, or chemical modification) at the Fc domain. Such modifications reduce the effector function (e.g., ADCC or ADCP) of antibodies or their antigen-binding fragments mediated by Fc receptors (e.g., FcγRI, FcγRII, or FcγRIII) and enhance the biological activity (e.g., agonist or antagonist activity) of antibodies or their antigen-binding fragments.

[0623] Anti-TNFRSF antibody or its antigen-binding fragment

[0624] Antibodies or antigen-binding fragments that specifically bind to TNFRSF member proteins (e.g., TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, or DCR3, etc.) are well known in the art. The antibody or its antigen-binding fragment may be an agonist antibody or its antigen-binding fragment that promotes or enhances the activation of TNFRSF member proteins upon binding. Alternatively, the antibody or its antigen-binding fragment may be an antagonist antibody or its antigen-binding fragment that inhibits or reduces the activation of TNFRSF member proteins upon binding.

[0625] Activating anti-TNFR2 antibodies or their antigen-binding fragments

[0626] In some embodiments, the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to TNFR2, comprising one or more CDRs having an amino acid sequence selected from Table 2 below or variants thereof having at most two conserved amino acid substitutions. For example, the antibody or its antigen-binding fragment may include CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 selected from Table 2.

[0627] Table 2. CDR sequences of agonistic anti-TNFR2 antibodies or their antigen-binding fragments.

[0628]

[0629]

[0630]

[0631]

[0632]

[0633]

[0634]

[0635]

[0636]

[0637]

[0638]

[0639]

[0640]

[0641]

[0642]

[0643]

[0644]

[0645]

[0646]

[0647]

[0648]

[0649]

[0650]

[0651]

[0652]

[0653] CDR-H1 of agonist antibodies that specifically bind to TNFR2 or their antigen-binding fragments can tolerate a wide range of amino acid modifications. See, for example, International Patent Application Publication No. WO 2021 / 231922, which is incorporated herein by reference. In some embodiments, the agonist antibody that specifically binds to TNFR2 or its antigen-binding fragment comprises CDR-H1 having the amino acid sequence GXTFXXY (SEQ ID NO: 1449), wherein each X is independently a naturally occurring amino acid. In some embodiments, the CDR-H1 has an amino acid sequence selected from the group consisting of: GYTFXXY (SEQ ID NO: 1450), GSTFXXY (SEQ ID NO: 1451), GTTFXXY (SEQ ID NO: 1452), GCTFXXY (SEQ ID NO: 1453), GNTFXXY (SEQ ID NO: 1454), GQTFXXY (SEQ ID NO: 1455), GXTFSXY (SEQ ID NO: 1456), GXTFCXY (SEQ ID NO: 1457), GXTFYXY (SEQ ID NO: 1458), GXTFNXY (SEQ ID NO: 1459), GXTFQXY (SEQ ID NO: 1460), and GXTFXDY (SEQ ID NO: 1461), wherein each X is independently a naturally occurring amino acid. In some embodiments, the CDR-H1 has the amino acid sequence GYTFTDY (SEQ ID NO: 1413) or a variant thereof having at most two conserved amino acid substitutions.In some embodiments, the agonistic antibody or antigen-binding fragment thereof that specifically binds to TNFR2 further comprises one or more of the following: CDR-H2, wherein the CDR-H2 has the amino acid sequence NPNYDS (SEQ ID NO: 1414), NYDSTS (SEQ ID NO: 1420), or variants thereof having at most two conserved amino acid substitutions; CDR-H3, wherein the CDR-H3 has the amino acid sequence GNSWYFDV (SEQ ID NO: 1415), or variants thereof having at most two conserved amino acid substitutions; CDR-L1, wherein the CDR-L1 has the amino acid sequence SASSSVRYMY (SEQ ID NO: 1416), or variants thereof having at most two conserved amino acid substitutions; CDR-L2, wherein the CDR-L2 has the amino acid sequence LTSNLAS (SEQ ID NO: 1417), or variants thereof having at most two conserved amino acid substitutions; and CDR-L3, wherein the CDR-L3 has the amino acid sequence QQWSSNPLT (SEQ ID NO: 14 ... 1418) or a variant thereof having at most two conserved amino acid substitutions.

[0654] Additional agonistic antibodies or their antigen-binding fragments that specifically bind to TNFR2 may include one or more of the following:

[0655] (a) CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of GYTFTDYNLD (SEQ ID NO: 1425), DYIMH (SEQ ID NO: 1426) or variants thereof having at most two conserved amino acid substitutions;

[0656] (b) CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of DINPNYDS (SEQ ID NO: 1427), WVDPEYGSTDYAEKFKK (SEQ ID NO: 1428) or variants thereof having at most two conserved amino acid substitutions;

[0657] (c) CDR-H3, wherein the CDR-H3 comprises the amino acid sequence of TSYSQKFRG (SEQ ID NO: 1429), DDGSYSPFEDY (SEQ ID NO: 1430) or variants thereof having at most two conserved amino acid substitutions;

[0658] (d) CDR-L1, wherein the CDR-L1 comprises an amino acid sequence selected from the group consisting of: SASSSVRYMYWY (SEQ ID NO: 1431), KASENVVTYVS (SEQ ID NO: 1432), RSSQSLVHSNGNTYLH (SEQ ID NO: 1433), KASENVVTYVS (SEQ ID NO: 1434), QASQNINKYIA (SEQ ID NO: 1435), QNINKY (SEQ ID NO: 1436), and variants thereof having at most two conserved amino acid substitutions;

[0659] (e) CDR-L2, wherein the CDR-L2 comprises an amino acid sequence selected from the group consisting of: LTSNLAS (SEQ ID NO: 1437), VTSNLAS (SEQ ID NO: 1438), LTSNLGS (SEQ ID NO: 1439), GASNRTY (SEQ ID NO: 1440), IKVSNRFS (SEQ ID NO: 1441), DPEYGS (SEQ ID NO: 1442), YTSTLES (SEQ ID NO: 1443), YTS, and variants thereof having at most two conserved amino acid substitutions; and

[0660] (f) CDR-L3, wherein the CDR-L3 comprises an amino acid sequence selected from the group consisting of: QQRSNWP (SEQ ID NO: 1445), GQGYSYPYT (SEQ ID NO: 1446), SQTTHVPPT (SEQ ID NO: 1447), LQYVNLIT (SEQ ID NO: 1448), and variants thereof having at most two conserved amino acid substitutions.

[0661] In some embodiments, the antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to TNFR2, comprising VH and / or VL, wherein the VH and / or the VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences selected from Table 3 below. In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment thereof disclosed in International Patent Application Publications No. WO 2020 / 102739, WO 2021 / 231922, WO 2020 / 061210, WO 2020 / 180712, WO 2021 / 055253, WO 2022 / 003690, WO 2020 / 089473, WO 2021 / 200840, or WO 2023 / 228082, which are incorporated herein by reference.

[0662] In some embodiments, the antibody or its antigen-binding fragment may be any antibody or its antigen-binding fragment disclosed in Examples 1, 3, 4, 10, 13 and 16 of International Patent Application Publication No. WO 2023 / 228082 and Tables 1-3, 8, 11 and 20, which are incorporated herein by reference. The antibody or its antigen-binding fragment of International Patent Application Publication No. WO 2023 / 228082 may be administered subcutaneously and / or at low doses, low dose ranges or low dosing frequencies to human subjects.

[0663] Table 3. VH and VL sequences of agonistic anti-TNFR2 antibodies or their antigen-binding fragments.

[0664]

[0665]

[0666]

[0667]

[0668]

[0669] Additional agonistic antibodies or their antigen-binding fragments that specifically bind to TNFR2 may include one or more of the following:

[0670] (a) CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of GYTFTDYNL (SEQ ID NO: 7) or a variant thereof having at most two conserved amino acid substitutions;

[0671] (b) CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of INPNYDST (SEQ ID NO: 8) or a variant thereof having at most two conserved amino acid substitutions;

[0672] (c) CDR-H3, wherein the CDR-H3 comprises the amino acid sequence of CARGNSWYFDV (SEQ ID NO: 9) or a variant thereof having at most two conserved amino acid substitutions;

[0673] (d) CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of SSVRY (SEQ ID NO: 10) or a variant thereof having at most two conserved amino acid substitutions;

[0674] (e) CDR-L2, wherein CDR-L2 comprises the amino acid sequence of LTS or a variant thereof having at most two conserved amino acid substitutions; and

[0675] (f) CDR-L3, which comprises the amino acid sequence of CQQWSSNPLT (SEQ ID NO: 12) or a variant thereof having at most two conserved amino acid substitutions.

[0676] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0677] (a) VH, said VH having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to any of SEQ ID NO: 1486-1490; and

[0678] (b) VL, wherein the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to any of SEQ ID NO: 1492-1497.

[0679] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0680] (a) VH, said VH having the amino acid sequence of any one of SEQ ID NO: 1486-1490; and

[0681] (b) VL, wherein the VL has the amino acid sequence of any one of SEQ ID NO: 1492-1497.

[0682] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0683] (a) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1488, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1495;

[0684] (b) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1488, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1497;

[0685] (c) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1489, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1496;

[0686] (d) VH and VL, wherein VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100%) to SEQ ID NO: 1490, and VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100%) to SEQ ID NO: 1496; or

[0687] (e) VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1490, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1497.

[0688] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0689] (a) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1488 and VL has the amino acid sequence of SEQ ID NO: 1495;

[0690] (b) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1488 and VL has the amino acid sequence of SEQ ID NO: 1497;

[0691] (c) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1489 and VL has the amino acid sequence of SEQ ID NO: 1496;

[0692] (d) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1490 and VL has the amino acid sequence of SEQ ID NO: 1496; or

[0693] (e) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1490 and VL has the amino acid sequence of SEQ ID NO: 1497.

[0694] In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1488, and the VL has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1495.

[0695] In some embodiments, the antibody or its antigen-binding fragment comprises VH and VL, wherein the VH has the amino acid sequence of SEQ ID NO: 1488 and the VL has the amino acid sequence of SEQ ID NO: 1495.

[0696] In some embodiments, the antibody or its antigen-binding fragment is a human antibody or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment is an IgG antibody or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment is a human IgG1 antibody, a human IgG2 antibody, a human IgG3 antibody, or a human IgG4 antibody or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment is a human IgG4 antibody or its antigen-binding fragment.

[0697] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0698] (a) a heavy chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to any of SEQ ID NO: 1498-1503; and

[0699] (b) A light chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to any of SEQ ID NO: 1504-1510.

[0700] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0701] (a) a heavy chain having the amino acid sequence of any one of SEQ ID NO: 1498-1503; and

[0702] (b) A light chain having an amino acid sequence of any one of SEQ ID NO: 1504-1510.

[0703] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0704] (a) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to that of SEQ ID NO: 1498, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to that of SEQ ID NO: 1504;

[0705] (b) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1501, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1508;

[0706] (c) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1501, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1510;

[0707] (d) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1502, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1509;

[0708] (e) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1503, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1509; or

[0709] (f) A heavy chain and a light chain, wherein the heavy chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1503, and the light chain has an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99% or 100% identical) to SEQ ID NO: 1510.

[0710] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0711] (a) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1498 and the light chain has the amino acid sequence of SEQ ID NO: 1504;

[0712] (b) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1501 and the light chain has the amino acid sequence of SEQ ID NO: 1508;

[0713] (c) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1501 and the light chain has the amino acid sequence of SEQ ID NO: 1510;

[0714] (d) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1502 and the light chain has the amino acid sequence of SEQ ID NO: 1509;

[0715] (e) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1503 and the light chain has the amino acid sequence of SEQ ID NO: 1509; or

[0716] (f) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1503 and the light chain has the amino acid sequence of SEQ ID NO: 1510.

[0717] In some embodiments, the antibody or its antigen-binding fragment comprises a heavy chain and a light chain, the heavy chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1501, and the light chain having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to SEQ ID NO: 1508.

[0718] In some embodiments, the antibody or its antigen-binding fragment comprises a heavy chain and a light chain, the heavy chain having the amino acid sequence of SEQ ID NO: 1501 and the light chain having the amino acid sequence of SEQ ID NO: 1508.

[0719] In some embodiments, the antibody or its antigen-binding fragment is a human IgG4 antibody or its antigen-binding fragment.

[0720] In some embodiments, the antibody or its antigen-binding fragment comprises one or more CDR sequences of antibody Ab1. Antibody Ab1 is a modified humanized IgG4 agonist anti-TNFR2 antibody comprising CDR-H1 of SEQ ID NO: 1413, CDR-H2 of SEQ ID NO: 1414 or 1420, and CDR-H3 of SEQ ID NO: 1415 in the variable weight (VH) domain, and CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 1416, 1417, and 1418 in the variable light (VL) domain.

[0721] Antagonistic anti-TNFR2 antibodies or their antigen-binding fragments

[0722] In some embodiments, the antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to TNFR2, comprising one or more CDRs having an amino acid sequence selected from Table 4 below or variants thereof having at most two conserved amino acid substitutions. For example, the antibody or its antigen-binding fragment may include CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 selected from Table 4.

[0723] Table 4. CDR sequences of antagonistic anti-TNFR2 antibodies or their antigen-binding fragments.

[0724]

[0725]

[0726]

[0727]

[0728]

[0729]

[0730]

[0731]

[0732]

[0733] In some embodiments, the antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to TNFR2, comprising VH and / or VL, wherein the VH and / or the VL have an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences selected from Table 5 below. In some embodiments, the antibody or antigen-binding fragment may be any antibody or its antigen-binding fragment disclosed in International Patent Application Publications Nos. WO 2020 / 089474, WO 2021 / 200840, WO 2021 / 249542, WO 2022 / 161425, WO 2022 / 166846, or WO 2022 / 267926, which are incorporated herein by reference.

[0734] Table 5. VH and VL sequences of antagonistic anti-TNFR2 antibodies or their antigen-binding fragments.

[0735]

[0736]

[0737]

[0738] Anti-TNFR2 antibodies or their antigen-binding fragments that are neither agonist nor antagonist

[0739] In some embodiments, the antibody or its antigen-binding fragment specifically binds to TNFR2, but is neither an agonist nor an antagonist. Such antibodies or their antigen-binding fragments are antibodies or their antigen-binding fragments comprising one or more CDRs having an amino acid sequence selected from those in Table 6 below or variants having at most two conserved amino acid substitutions. For example, the antibody or its antigen-binding fragment may include CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 selected from Table 6.

[0740] Table 6. CDR sequences of anti-TNFR2 antibodies or their antigen-binding fragments that are neither agonists nor antagonists.

[0741]

[0742] In some embodiments, the antibody or its antigen-binding fragment is neither an agonist nor an antagonist that specifically binds to TNFR2, and comprises VH and / or VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences selected from Table 7 below. In some embodiments, the antibody or antigen-binding fragment may be any antibody or its antigen-binding fragment disclosed in International Patent Application Publication No. WO 2022 / 003693, which is incorporated herein by reference.

[0743] Table 7. VH and VL sequences of antagonistic anti-TNFR2 antibodies or their antigen-binding fragments.

[0744]

[0745] Antibodies or their antigen-binding fragments that bind to other TNFRSF member proteins

[0746] Antibodies or antigen-binding fragments thereof that specifically bind to other TNFRSF member proteins (e.g., 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TNFR1, TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3, etc.) are also well known in the art. Each of the antibodies or antigen-binding fragments described herein can be formulated as a modified antibody or antigen-binding fragment comprising a modified Fc domain.

[0747] In some embodiments, the antibody or antigen-binding fragment may be from International Patent Application Publications No. WO2005 / 035584, WO 2012 / 032433, WO 2017 / 205745, WO 2018 / 017761, WO2018 / 098370, WO 2018 / 114748, WO 2018 / 114754, WO 2018 / 127473, WO2019 / 091436, WO 2019 / 140425, WO 2019 / 175125, WO 2019 / 196868, WO2020 / 007817, WO 2020 / 073131, etc. No. 2020 / 111913, No. WO 2021 / 030488, No. WO2021 / 089588, No. WO 2021 / 093753, No. WO 2021 / 101346, No. WO 2021 / 118246, No. WO2021 / 132746, No. WO No. 2021 / 167915, No. WO 2021 / 207827, No. WO 2022 / 039490, No. WO2022 / 148413, No. WO 2022 / 178114, No. WO 2022 / 200478, No. WO No. 2023 / 073225, No. WO2023 / 110788, No. WO Any antibody or antigen-binding fragment that specifically binds to 4-1BB as disclosed in WO 2023 / 134657 or WO 2023 / 174140 is incorporated herein by reference.

[0748] In some embodiments, the anti-4-1BB antibody or its antigen-binding fragment comprises one or more CDR sequences of utolumab (PF-05082566) having at most two conserved amino acid substitutions. In some embodiments, the anti-4-1BB antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of utolumab (PF-05082566). Utolumab (PF-05082566) and other anti-4-1BB antibodies are disclosed, for example, in WO 2012 / 032433; which is incorporated herein by reference.

[0749] In some embodiments, the anti-4-1BB antibody or its antigen-binding fragment comprises one or more CDR sequences of uroselumab (BMS-663513) having at most two conserved amino acid substitutions. In some embodiments, the anti-4-1BB antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of uroselumab (BMS-663513). Urelumab (BMS-663513) and other anti-4-1BB antibodies are disclosed, for example, in WO 2005 / 035584 and WO 2016 / 029073; which are incorporated herein by reference.

[0750] In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment that specifically binds to CD27 as disclosed in International Patent Application Publications No. WO2011 / 130434, WO 2010 / 001908, WO 2012 / 004367, WO 2015 / 016718, WO2019 / 196117 or WO 2021 / 087016, which is incorporated herein by reference.

[0751] In some embodiments, the anti-CD27 antibody or its antigen-binding fragment comprises one or more CDR sequences of variliterumab (CDX-1127) having at most two conserved amino acid substitutions. In some embodiments, the anti-CD27 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of variliterumab (CDX-1127). Variliterumab (CDX-1127) and other anti-CD27 antibodies are disclosed, for example, in WO 2011 / 130434; which is incorporated herein by reference.

[0752] In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment thereof that specifically binds to CD30 as disclosed in International Patent Application Publications No. WO2008 / 025020, WO 2017 / 066122, WO 2020 / 135559, WO 2007 / 040653, WO2023 / 057571, WO 2021 / 231568, WO 2021 / 091815, WO 2022 / 120084, WO2021 / 155129, WO 2023 / 076989, WO 03 / 104432, WO 97 / 17374, or WO 96 / 22384; which is incorporated herein by reference.

[0753] In some embodiments, the antibody or antigen-binding fragment may be from International Patent Application Publications No. WO2017 / 004016, WO 2018 / 185045, WO 2016 / 196314, WO 2017 / 040932, WO2017 / 184619, WO 2017 / 004006, WO 2016 / 023960, WO 2018 / 189220, WO2014 / 207064, WO 2016 / 069919, WO 2009 / 062054, WO 2012 / 075111, WO2020 / 070035, WO 2007 / 075326, etc. No. 2006 / 128103, No. WO 2017 / 205742, No. WO2018 / 011421, No. WO 2012 / 149356, No. WO 2014 / 070934, No. WO 2014 / 065402, No. WO2014 / 065403, No. WO No. 2005 / 063289, No. WO 2010 / 104761, No. WO 2021 / 081303, No. WO2020 / 253722, No. WO 2020 / 154335, No. WO 2019 / 093342, No. WO No. 2018 / 017763, No. WO2013 / 034904, No. WO No. 2019 / 106608, No. WO 2020 / 207470, No. WO 2018 / 088850, No. WO2018 / 085533, No. WO 2021 / 183428, No. WO 2021 / 140222, No. WO No. 2019 / 057792, No. WO2012 / 041635, No. WO 2018 / 178046, No. WO 2020 / 014974, No. WO 2020 / 065409, No. WO2021 / 222188, No. WO 2022 / 078357, No. WO No. 2020 / 230899, WO No. 2020 / 230901, No. WO2019 / 204756, No. WO 2021 / 239968, No. WO 2022 / 133123, No. WO 2022 / 101458, No. WO2022 / 251311, No. WO 2023 / 274007, No. WO No. 2022 / 243261, No. WO 2023 / 284714, No. WO2023 / 046037, No. WO 03 / 040170, No. WO 2023 / 274201, No. WO 2023 / 198194, No. WO2023 / 020475, No. WOAny antibody or antigen-binding fragment that specifically binds to CD40 disclosed in WO 2023 / 125652, WO 2022 / 037662, WO 2021 / 243422 or WO2023 / 052581 is incorporated herein by reference.

[0754] In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises one or more CDR sequences of CDX-1140 having at most two conserved amino acid substitutions. In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of CDX-1140. CDX-1140 and other anti-CD40 antibodies are disclosed, for example, in WO 2017 / 184619 and Vitale et al. Cancer Immunol. Immunother. 68:233-245, 2019; which are incorporated herein by reference.

[0755] In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises one or more CDR sequences of SEA-CD40 having at most two conserved amino acid substitutions. In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of SEA-CD40. SEA-CD40 and other anti-CD40 antibodies are disclosed, for example, in WO 2016 / 069919; incorporated herein by reference.

[0756] In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises one or more CDR sequences of RO7009789 having at most two conserved amino acid substitutions. In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of RO7009789. RO7009789 and other anti-CD40 antibodies are disclosed, for example, in US 7,338,660; WO 03 / 040170; and Vonderheide et al. J. Clin. Oncol. 25:876,2007; which are incorporated herein by reference.

[0757] In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises one or more CDR sequences of JNJ-64457107 (ADC1013) having at most two conserved amino acid substitutions. In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of JNJ-64457107 (ADC1013). JNJ-64457107 (ADC1013) and other anti-CD40 antibodies are disclosed, for example, in WO 2016 / 023960 and Mangsbo et al. Clin. Cancer Res. 21:1115-1126, 2015; which are incorporated herein by reference.

[0758] In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises one or more CDR sequences of APX-005M having at most two conserved amino acid substitutions. In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of APX-005M. APX-005M and other anti-CD40 antibodies are disclosed, for example, in WO 2014 / 070934 and WO 2018 / 085533; which are incorporated herein by reference.

[0759] In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises one or more CDR sequences of Chi Lob 7 / 4 having at most two conserved amino acid substitutions. In some embodiments, the anti-CD40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of Chi Lob 7 / 4. Chi Lob 7 / 4 and other anti-CD40 antibodies are disclosed, for example, in US 2009 / 0074711 and Johnson et al. Clin. Cancer Res. 21:1321-1328,2015; which are incorporated herein by reference.

[0760] In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment that specifically binds to DR6 as disclosed in International Patent Application Publication No. WO2010 / 062904, which is incorporated herein by reference.

[0761] In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment that specifically binds to EDAR as disclosed in International Patent Application Publication No. WO2010 / 113117, which is incorporated herein by reference.

[0762] In some embodiments, the antibody or antigen-binding fragment may be from International Patent Application Publications Nos. WO2017 / 096189, WO 2016 / 054638, WO 2017 / 096276, WO 2017 / 087678, WO2015 / 187835, WO 2015 / 184099, WO 2020 / 218951, WO 2018 / 094300, WO2006 / 105021, WO 2018 / 185618, WO 2018 / 017889, WO 2020 / 163646, WO2011 / 028683, WO 2018 / 158658, etc. Any antibody or antigen-binding fragment that specifically binds to HVEM disclosed in WO 2015 / 031667, WO 2009 / 009116, WO 2017 / 214548, WO 2018 / 091739, WO 2018 / 018039, WO 2020 / 108636, WO 2019 / 201301, WO 2019 / 184898, WO 2022 / 240161, WO 2023 / 175614 or WO 2021 / 178814; incorporated herein by reference.

[0763] In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment thereof that specifically binds to GITR as disclosed in International Patent Application Publications No. WO2020 / 222235, WO 2014 / 184360, WO 2015 / 031667, WO 2022 / 208505, WO2022 / 197866, or WO 2021 / 160266, WO 2006 / 105021, WO 2011 / 028683, WO2015 / 026684, WO 2015 / 184099, or WO 2016 / 196792; which is incorporated herein by reference.

[0764] In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises one or more CDR sequences of TRX-518 having at most two conserved amino acid substitutions. In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of TRX-518. TRX-518 and other anti-GITR antibodies are disclosed, for example, in US 7,812,135; US 8,388,967; US 9,028,823; WO 2006 / 105021; and Ponte J et al. Clinical Immunology 135:S96, 2010; which are incorporated herein by reference.

[0765] In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises one or more CDR sequences of MK-4166 or MK-1248 having at most two conserved amino acid substitutions. In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of MK-4166 or MK-1248. MK-4166, MK-1248, and other anti-GITR antibodies are disclosed, for example, in US 8,709,424; WO 2011 / 028683; WO 2015 / 026684; and Mahne et al. Cancer Res. 77:1108-1118, 2017; which are incorporated herein by reference.

[0766] In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises one or more CDR sequences of GWN-323 having at most two conserved amino acid substitutions. In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of GWN-323. GWN-323 and other anti-GITR antibodies are disclosed, for example, in Piha-Paul et al. J. Immunother. Cancer 9:e002863, 2021; incorporated herein by reference.

[0767] In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises one or more CDR sequences of INCAGN01876 having at most two conserved amino acid substitutions. In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of INCAGN01876. INCAGN01876 and other anti-GITR antibodies are disclosed, for example, in US 10,155,818 and WO 2015 / 184099; which are incorporated herein by reference.

[0768] In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises one or more CDR sequences of BMS-986156 having at most two conserved amino acid substitutions. In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of BMS-986156. BMS-986156 and other anti-GITR antibodies are disclosed, for example, in US 9,228,016 and WO 2016 / 196792; which are incorporated herein by reference.

[0769] In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises one or more CDR sequences of AMG-228 having at most two conserved amino acid substitutions. In some embodiments, the anti-GITR antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of AMG-228. AMG-228 and other anti-GITR antibodies are disclosed, for example, in US 9,464,139 and WO 2015 / 031667; which are incorporated herein by reference.

[0770] In some embodiments, the antibody or antigen-binding fragment may be from International Patent Application Publications No. WO2018 / 177220, WO 2017 / 063162, WO 2018 / 112346, WO 2017 / 096281, WO2017 / 096182, WO 2016 / 179517, WO 2018 / 202649, WO 2010 / 096418, WO2013 / 008171, WO 2013 / 068563, WO 2015 / 095423, WO 2015 / 153514, WO2016 / 073380, WO 2016 / 081384, etc. No. 2016 / 200836, No. WO 2017 / 130076, No. WO2019 / 086497, No. WO 2018 / 031490, No. WO 2018 / 017888, No. WO 2018 / 002339, No. WO2020 / 103836, No. WO No. 2007 / 062245, No. WO 2021 / 098851, No. WO 2019 / 214624, No. WO2013 / 028231, No. WO 2019 / 089921, No. WO 2012 / 027328, No. WO No. 2020 / 151761, No. WO2019 / 100320, No. WO Any antibody or antigen-binding fragment that specifically binds to OX40 disclosed in WO 2020 / 259667, WO 2021 / 030488, WO 2023 / 109900, WO 2022 / 258015, WO 2022 / 178114, WO 2023 / 089079, WO 2020 / 063668, WO 2023 / 143597, WO 2020 / 119789, WO 2020 / 119793, WO 2020 / 119792, WO 2022 / 002009, or WO 2022 / 262749; incorporated herein by reference.

[0771] In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises one or more CDR sequences of taliximab (MEDI0562) having at most two conserved amino acid substitutions. In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of taliximab (MEDI0562). Taliximab (MEDI0562) and other anti-OX40 antibodies are disclosed, for example, in WO 2015 / 095423; WO 2015 / 153514; WO2016 / 073380; and WO 2016 / 081384; which are incorporated herein by reference.

[0772] In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises one or more CDR sequences of PF-04518600 having at most two conserved amino acid substitutions. In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of PF-04518600. PF-04518600 and other anti-OX40 antibodies are disclosed, for example, in WO 2017 / 130076; which is incorporated herein by reference.

[0773] In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises one or more CDR sequences of BMS-986178 having at most two conserved amino acid substitutions. In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of BMS-986178. BMS-986178 and other anti-OX40 antibodies are disclosed, for example, in WO 2019 / 089921; which is incorporated herein by reference.

[0774] In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises one or more CDR sequences of MOXR-0916 having at most two conserved amino acid substitutions. In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of MOXR-0916. MOXR-0916 and other anti-OX40 antibodies are disclosed, for example, in WO 2016 / 200836; which is incorporated herein by reference.

[0775] In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises one or more CDR sequences of GSK-3174998 having at most two conserved amino acid substitutions. In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of GSK-3174998.

[0776] In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises one or more CDR sequences of INCAGN01949 having at most two conserved amino acid substitutions. In some embodiments, the anti-OX40 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of INCAGN01949. INCAGN01949 and other anti-OX40 antibodies are disclosed, for example, in WO 2016 / 179517; which is incorporated herein by reference.

[0777] In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment that specifically binds to RANK as disclosed in International Patent Application Publication No. WO2020 / 113274, which is incorporated herein by reference.

[0778] In some embodiments, the antibody or antigen-binding fragment may be any antibody or antigen-binding fragment that specifically binds to TRAIL receptors (e.g., TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), or TRAIL-R4) as disclosed in International Patent Application Publications No. WO2016 / 111344, WO 2017 / 028279, WO 2007 / 027713, or WO 2016 / 074245, which is incorporated herein by reference.

[0779] Agonistant anti-GITR antibodies and agonist anti-TNFR2 antibodies

[0780] GITR is a major immune receptor expressed on T cells and natural killer cells and activated by its ligand GITRL. GITRL is normally expressed on antigen-presenting cells and endothelial cells. Upon activation, GITR stimulates both adaptive and innate immunity. GITR may be involved in immune responses to infection and tumors, and its activation may cause inflammation and graft-versus-host disease. Agonisttic anti-GITR antibodies can be used to treat cancer, but are harmful to patients with autoimmune diseases because they can activate effector T cells while suppressing Treg cells.

[0781] GITR plays an opposite role to TNFR2 in the immune response. TNFR2 is an immune receptor whose expression in adults is very limited. Typically, only a restricted subset of Treg cells express TNFR2. TNFR2 expression in this subset of Treg cells has identified a subset with potent immunosuppressive properties. TNFR2 functions to calm the immune response, reduce inflammation, and suppress autoimmunity. Excessive activation of TNFR2 may contribute to cancer due to increased immune tolerance.

[0782] Furthermore, GITR differs structurally from TNFR2. GITR contains only two extracellular subdomains, while TNFR2 contains four extracellular subdomains (CRD1, CRD2, CRD3, and CRD4). Antibodies targeting TNFR2 are known to exhibit the highest agonistic activity upon binding to CRD1 and / or CRD2. The two extracellular subdomains of GITR correspond to CRD3 and CRD4 of TNFR2. Given their distinct and separate functions, anti-GITR antibodies will not be used to inform the design of agonistic TNFR2 antibodies.

[0783] Anti-TNFSF antibody or its antigen-binding fragment

[0784] Antibodies or antigen-binding fragments thereof that specifically bind to TNFSF member proteins (e.g., TNF-α, TNF-β, lymphotoxin-β (LT-β), CD40L, FasL, CD30L, 4-1BBL, CD27L, OX40L, TRAIL, LIGHT, RANKL, TWEAK, APRIL, BAFF, VEGI, EDA-A1, EDA-A2, and GITRL, etc.) are well known in the art. As described herein, each of the antibodies or antigen-binding fragments described below can be formulated as a modified antibody or antigen-binding fragment comprising a modified Fc domain. In some embodiments, the antibody or antigen-binding fragment can be any antibody or antigen-binding fragment that specifically binds to TRAIL.

[0785] CDRs of agonistic antibodies or their antigen-binding fragments that bind to TNFRSF member proteins

[0786] CDR-H1 sequences of agonistic antibodies or antigen-binding fragments thereof that bind to TNFRSF member proteins (e.g., TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3) are particularly important in determining the binding specificity and agonistic activity of such antibodies or their antigen-binding fragments. Efficient signal transduction of TNFRSF member proteins requires the formation of homotrimeric receptor structures. Antibodies or their antigen-binding fragments containing specific CDR-H1 sequences that specifically bind to one or more amino acid residues within the trimer interface of TNFRSF member proteins can stabilize the trimer structure, thereby acting as agonists of said TNFRSF member proteins.

[0787] In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains X 1 X 2 X 2 X 1 X 2 X 3 X 2 The amino acid sequence of JJJ, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X 2 Independently, it is Y, S, T, C, N, or Q; X 3It is either D or E; and each J is independently a naturally occurring amino acid or is not present. For example, each amino acid residue of a specific CDR-H1 sequence of an agonist TNFR2 antibody or its antigen-binding fragment (i.e., GYTFTDY (SEQ ID NO: 1413)) can be replaced by an amino acid with similar physicochemical properties (e.g., polarity, charge, acidity or basicity), as shown in Table 8 below.

[0788] Table 8. Acceptable amino acid substitutions for exemplary CDR-H1 sequences of agonistic TNFR2 antibodies or their antigen-binding fragments.

[0789]

[0790] In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains GYTFTZ. 1 Z 2 The amino acid sequence of JJJ (SEQ ID NO: 1480), where Z 1 It is D or T; Z 2 It is Y, F, or L; and each J is independently a naturally occurring amino acid or is absent. Antibodies comprising CDR-H1 including the amino acid sequence of SEQ ID NO: 1480 are known in the art to bind to human TNFR2 and exert agonistic activity. For example, International Patent Application Publication No. WO 2020 / 061210 teaches that antibodies comprising CDR-H1 having the amino acid sequence GYTFTTF (SEQ ID NO: 31), GYTFTTFGMS (SEQ ID NO: 37), or GYTFTTFG (SEQ ID NO: 73) are agonistic antibodies that bind to human TNFR2. International Patent Application Publication No. WO 2021 / 141907 teaches that antibodies comprising CDR-H1 having the amino acid sequence GYTFTDYY (SEQ ID NO: 325) are agonistic antibodies that bind to human TNFR2. International Patent Application Publication No. WO 2022 / 003690 teaches that an antibody comprising CDR-H1 having the amino acid sequence GYTFTDLG (SEQ ID NO: 481) is an agonist antibody that binds to human TNFR2. In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0791] In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413) or a variant thereof having one or more conserved amino acid substitutions. In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413) or a variant thereof having at most two conserved amino acid substitutions. In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413). In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0792] In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains X 1 X 2 X 2 X 3 X 1 The amino acid sequence, where each X 1 Independently, it is either D or E; each X 2 Independently, it is Y, S, T, C, N, or Q; and X 3 It is L, A, V, G, I, M, W, F, or P. For example, each amino acid residue of a specific CDR-H1 sequence (i.e., DYNLD (SEQ ID NO: 1541)) of an agonist TNFR2 antibody or its antigen-binding fragment can be replaced by an amino acid with similar physicochemical properties (e.g., polarity, charge, acidity, or basicity), as shown in Table 9 below.

[0793] Table 9. Acceptable amino acid substitutions for exemplary CDR-H1 sequences of agonistic TNFR2 antibodies or their antigen-binding fragments.

[0794]

[0795] In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of DYNLD (SEQ ID NO: 1541) or a variant thereof having one or more conserved amino acid substitutions. In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of DYNLD (SEQ ID NO: 1541) or a variant thereof having at most two conserved amino acid substitutions. In some embodiments, the CDR-H1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of DYNLD (SEQ ID NO: 1541). In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0796] Agonisttic antibodies or their antigen-binding fragments that bind to TNFRSF member proteins (e.g., TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3) can tolerate significant variations in CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3. Amino acid substitutions (e.g., conserved amino acid substitutions) at CDR-H2, CDR-H3, CDR-L1, CDR-L2, or CDR-L3 of agonist antibodies or their antigen-binding fragments that bind to TNFRSF member proteins do not affect the binding specificity and agonist activity of these antibodies or their antigen-binding fragments.

[0797] In some embodiments, the CDR-H2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains X 1 X 2 X 3 X 2 X 3 X 3 X 1 X 3 X 3 X 3 X 3 X 3 X 3 X 4 X 2 X 4X 2 The amino acid sequence, where each X 1 Independently, it is either D or E; each X 2 Independently, it is I, A, V, L, G, M, W, F, or P; each X 3 Independently, it is N, S, T, C, Y, or Q; and each X 4 Independently, it is K, R, or H. For example, each amino acid residue of a specific CDR-H2 sequence of an agonist TNFR2 antibody or its antigen-binding fragment (i.e., DINPNYDSTSYSQKFRG (SEQ ID NO: 1542)) can be replaced by an amino acid with similar physicochemical properties (e.g., polarity, charge, acidity, or basicity), as shown in Table 10 below.

[0798] Table 10. Acceptable amino acid substitutions for exemplary CDR-H2 sequences of agonistic TNFR2 antibodies or their antigen-binding fragments.

[0799]

[0800] In some embodiments, the CDR-H2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of DINPNYDSTSYSQKFRG (SEQ ID NO: 1542) or a variant thereof having one or more conserved amino acid substitutions. In some embodiments, the CDR-H2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of DINPNYDSTSYSQKFRG (SEQ ID NO: 1542) or a variant thereof having at most two conserved amino acid substitutions. In some embodiments, the CDR-H2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of DINPNYDSTSYSQKFRG (SEQ ID NO: 1542). In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure further comprises one or more of the following: CDR-H1, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0801] In some embodiments, the CDR-H3 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains X 1 X 2 X 2 X 1 X 2 X 1 X 3 X 1 The amino acid sequence, where each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; each X2 Independently, it is N, S, T, C, Y, or Q; and X 3 It is D or E. For example, each amino acid residue of a specific CDR-H3 sequence of an agonist TNFR2 antibody or its antigen-binding fragment (i.e., GNSWYFDV (SEQ ID NO: 1543)) can be replaced by an amino acid with similar physicochemical properties (e.g., polarity, charge, acidity or basicity), as shown in Table 11 below.

[0802] Table 11. Acceptable amino acid substitutions for exemplary CDR-H3 sequences of agonistic TNFR2 antibodies or their antigen-binding fragments.

[0803]

[0804] In some embodiments, the CDR-H3 of the agonist TNFR2 antibody or its antigen-binding fragment of this disclosure comprises the amino acid sequence of GNSWYFDV (SEQ ID NO: 1543) or a variant thereof having one or more conserved amino acid substitutions. In some embodiments, the CDR-H3 of the agonist TNFR2 antibody or its antigen-binding fragment of this disclosure comprises the amino acid sequence of GNSWYFDV (SEQ ID NO: 1543) or a variant thereof having at most two conserved amino acid substitutions. In some embodiments, the CDR-H3 of the agonist TNFR2 antibody or its antigen-binding fragment of this disclosure comprises the amino acid sequence of GNSWYFDV (SEQ ID NO: 1543). In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of this disclosure further comprises one or more of the following: CDR-H1, CDR-H2, CDR-L1, CDR-L2, and CDR-L3 shown in Table 2.

[0805] In some embodiments, the CDR-L1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains X 1 X 2 X 1 X 1 X 1 X 2 X 3 X 4 X 1 X 1 The amino acid sequence, where each X 1 Independently, it is S, Y, T, C, N, or Q; each X 2 Independently, it is A, G, V, L, I, M, W, F, or P; X 3 It is R, H, or K; and X 4It is Y, A, V, I, L, M, F, or W. For example, each amino acid residue of a specific CDR-L1 sequence of an agonist TNFR2 antibody or its antigen-binding fragment (i.e., SASSSVRYNY (SEQ ID NO: 1544)) can be replaced by an amino acid with similar physicochemical properties (e.g., polarity, charge, acidity, or basicity), as shown in Table 12 below.

[0806] Table 12. Acceptable amino acid substitutions for exemplary CDR-L1 sequences of agonistic TNFR2 antibodies or their antigen-binding fragments.

[0807]

[0808] In some embodiments, the CDR-L1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of SASSSVRYNY (SEQ ID NO: 1544) or a variant thereof having one or more conserved amino acid substitutions. In some embodiments, the CDR-L1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of SASSSVRYNY (SEQ ID NO: 1544) or a variant thereof having at most two conserved amino acid substitutions. In some embodiments, the CDR-L1 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of SASSSVRYNY (SEQ ID NO: 1544). In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L2, and CDR-L3 shown in Table 2.

[0809] In some embodiments, the CDR-L2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is L, A, V, G, I, M, W, F, or P; and each X 2 Independently, it is T, S, C, Y, N, or Q. For example, each amino acid residue of a specific CDR-L2 sequence (i.e., LTSNLAS (SEQ ID NO: 1545)) of an agonist TNFR2 antibody or its antigen-binding fragment can be replaced by an amino acid with similar physicochemical properties (e.g., polarity, charge, acidity, or basicity), as shown in Table 13 below.

[0810] Table 13. Acceptable amino acid substitutions for exemplary CDR-L2 sequences of agonistic TNFR2 antibodies or their antigen-binding fragments.

[0811]

[0812] In some embodiments, the CDR-L2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of LTSNLAS (SEQ ID NO: 1545) or a variant thereof having one or more conserved amino acid substitutions. In some embodiments, the CDR-L2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of LTSNLAS (SEQ ID NO: 1545) or a variant thereof having at most two conserved amino acid substitutions. In some embodiments, the CDR-L2 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of LTSNLAS (SEQ ID NO: 1545). In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L3 shown in Table 2.

[0813] In some embodiments, the CDR-L3 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure contains X 1 X 2 X 2 X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, where each X 1 Independently, it is P, A, V, L, I, M, W, F, or G; and each X 2 Independently, it is Q, S, T, C, N, or Y. For example, each amino acid residue of a specific CDR-L3 sequence (i.e., PQQWSSNPLT (SEQ ID NO: 1546)) of an agonist TNFR2 antibody or its antigen-binding fragment can be replaced by an amino acid with similar physicochemical properties (e.g., polarity, charge, acidity, or basicity), as shown in Table 14 below.

[0814] Table 14. Acceptable amino acid substitutions for exemplary CDR-L3 sequences of agonistic TNFR2 antibodies or their antigen-binding fragments.

[0815]

[0816] In some embodiments, the CDR-L3 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of PQQWSSNPLT (SEQ ID NO: 1546) or a variant thereof having one or more conserved amino acid substitutions. In some embodiments, the CDR-L3 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of PQQWSSNPLT (SEQ ID NO: 1546) or a variant thereof having at most two conserved amino acid substitutions. In some embodiments, the CDR-L3 of the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure comprises the amino acid sequence of PQQWSSNPLT (SEQ ID NO: 1546). In some embodiments, the agonist TNFR2 antibody or its antigen-binding fragment of the present disclosure further comprises one or more of the following: CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L2 shown in Table 2.

[0817] In some embodiments, any of the agonist TNFR2 antibodies or antigen-binding fragments disclosed herein may be administered subcutaneously to human subjects. In some embodiments, any of the agonist TNFR2 antibodies or antigen-binding fragments disclosed herein may be administered to human subjects at low doses.

[0818] Cross-reactivity of anti-TNFSF antibodies or their antigen-binding fragments with soluble TNFRSF member proteins

[0819] The anti-TNFRSF antibodies or antigen-binding fragments thereof disclosed herein (e.g., antibodies or antigen-binding fragments that bind to members of the TNFRSF protein family (such as TNFR2, TNFR1, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3)) may have low cross-reactivity with soluble forms of TNFRSF member proteins. For example, the TNFR2 antibody or antigen-binding fragment of this disclosure may have low cross-reactivity with soluble TNFR2. Such anti-TNFRSF antibodies or antigen-binding fragments thereof may have superior biological activity compared to antibodies or antigen-binding fragments that cross-react with soluble forms of TNFRSF member proteins.

[0820] It is known in the art that agonistic antibodies or antigen-binding fragments thereof that bind to TNFRSF member proteins can activate the expression of soluble forms of TNFRSF member proteins. If such agonistic antibodies or antigen-binding fragments thereof cross-react with soluble forms of TNFRSF member proteins, the soluble forms of TNFRSF member proteins can act as decoys to neutralize the agonistic antibodies or antigen-binding fragments thereof, resulting in reduced agonistic activity of the antibodies or antigen-binding fragments thereof, and potentially requiring increased dosage and / or frequency of administration of the antibodies or antigen-binding fragments thereof to achieve a therapeutic effect in subjects.

[0821] In some embodiments, the modified antibody or its antigen-binding fragment has less than about 90% (e.g., less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10%) cross-reactivity with the soluble TNFRSF member protein. In some embodiments, the modified antibody or its antigen-binding fragment has less than 50% cross-reactivity with the soluble TNFRSF member protein. The cross-reactivity of the antibody or its antigen-binding fragment with the antigen can be determined using methods known in the art, such as by assessing the percentage of homology between the antigen sequence of the antibody-bound protein and the sequence of a second protein. A high percentage of homology indicates that the antibody is highly likely to cross-react with the second protein.

[0822] Due to the low cross-reactivity of the anti-TNFRSF antibody or antigen-binding fragment thereof disclosed herein with soluble forms of TNFRSF member proteins, the anti-TNFRSF antibody or antigen-binding fragment thereof disclosed herein can be used at low doses (e.g., doses lower than typical (or art-recognized) dose levels of the corresponding antibody or antigen-binding fragment thereof) (e.g., administered to human subjects). In some embodiments, the antibody or antigen-binding fragment thereof is administered once weekly at a dose of about 0.8 mg / kg to about 1.2 mg / kg (e.g., about 0.8 mg / kg, about 0.85 mg / kg, about 0.9 mg / kg, about 0.95 mg / kg, about 1.0 mg / kg, about 1.05 mg / kg, about 1.1 mg / kg, about 1.15 mg / kg, or about 1.2 mg / kg). In some embodiments, the antibody or its antigen-binding fragment is administered every two weeks at a dose of about 1.65 mg / kg to about 2.5 mg / kg (e.g., about 1.65 mg / kg, about 1.7 mg / kg, about 1.75 mg / kg, about 1.8 mg / kg, about 1.85 mg / kg, about 1.9 mg / kg, about 1.95 mg / kg, about 2.0 mg / kg, about 2.05 mg / kg, about 2.1 mg / kg, about 2.15 mg / kg, about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg, or about 2.5 mg / kg). In some embodiments, the antibody or its antigen-binding fragment is administered once every three weeks at a dose of about 2.2 mg / kg to about 3.0 mg / kg (e.g., about 2.2 mg / kg, about 2.25 mg / kg, about 2.3 mg / kg, about 2.35 mg / kg, about 2.4 mg / kg, about 2.45 mg / kg, about 2.5 mg / kg, about 2.55 mg / kg, about 2.6 mg / kg, about 2.65 mg / kg, about 2.7 mg / kg, about 2.75 mg / kg, about 2.8 mg / kg, about 2.85 mg / kg, about 2.9 mg / kg, about 2.95 mg / kg, or about 3.0 mg / kg). In some embodiments, the antibody or its antigen-binding fragment is administered once every four weeks at a dose of about 2.5 mg / kg to about 4.2 mg / kg (e.g., about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4.0 mg / kg, about 4.1 mg / kg, or about 4.2 mg / kg).

[0823] Anti-CD28 antibodies and anti-ICOS antibodies or their antigen-binding fragments

[0824] Antibodies or antigen-binding fragments thereof that specifically bind to CD28 or ICOS are well known in the art. As described herein, each of the antibodies or antigen-binding fragments described below can be formulated as a modified antibody or antigen-binding fragment thereof comprising a modified Fc domain.

[0825] In some embodiments, the antibody or antigen-binding fragment may be from International Patent Application Publications No. WO2020 / 127618, WO 2020 / 076853, WO 2016 / 185016, WO 2020 / 132066, WO2018 / 177966, WO 2019 / 246514, WO 2010 / 007376, WO 2020 / 198009, WO2021 / 259890, WO 2021 / 155380, WO 2020 / 127628, WO 2021 / 260064, WO2015 / 198147, WO 2022 / 061098, etc. No. 2011 / 101791, No. WO 2020 / 153605, No. WO2006 / 050949, No. WO 2020 / 132024, No. WO 2016 / 146702, No. WO 2012 / 080351, No. WO2019 / 240934, No. WO No. 2011 / 042891, No. WO 2022 / 253867, No. WO 2022 / 269019, No. WO2023 / 143547, No. WO 01 / 88159, No. WO 2023 / 068382, No. WO No. 2020 / 210392, No. WO2023 / 126445, No. WO Any antibody or antigen-binding fragment that specifically binds to CD28 disclosed in WO 2023 / 215498, WO 2023 / 193239, WO 2022 / 170033, WO 2022 / 094299, WO 2023 / 215799, WO 2022 / 232392, WO 2023 / 180303, WO 02 / 051871, or WO 2023 / 031943 is incorporated herein by reference.

[0826] In some embodiments, the anti-CD28 antibody or its antigen-binding fragment comprises one or more CDR sequences of cilalizumab (TAB-08) having at most two conserved amino acid substitutions. In some embodiments, the anti-CD28 antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequences of cilalizumab (TAB-08). Cilalizumab (TAB-08) and other anti-CD28 antibodies are disclosed, for example, in US 2006 / 0009382 and Beyersdorf et al. Ann. Rheum. Dis. 64 Supplement 4:iv91-95, 2005; which are incorporated herein by reference.

[0827] In some embodiments, the antibody or antigen-binding fragment may be from International Patent Application Publications Nos. WO2016 / 120789, WO 2016 / 154177, WO 2018 / 029474, WO 2018 / 025221, WO2019 / 122049, WO 2012 / 131004, WO 2020 / 102233, WO 2021 / 043961, WO2019 / 229614, WO 2019 / 229613, WO 2019 / 171294, WO 2020 / 260326, WO2020 / 031087, WO 2021 / 046293, etc. Any antibody or antigen-binding fragment that specifically binds to ICOS as disclosed in WO 2022 / 254227, WO 2020 / 086476, WO 2022 / 098910, WO 2021 / 229032 or WO 2021 / 216417 is incorporated herein by reference.

[0828] In some embodiments, the anti-ICOS antibody or its antigen-binding fragment comprises one or more CDR sequences of GSK-3359609 having at most two conserved amino acid substitutions. In some embodiments, the anti-ICOS antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of GSK-3359609. GSK-3359609 and other anti-ICOS antibodies are disclosed, for example, in WO 2016 / 120789; which is incorporated herein by reference.

[0829] In some embodiments, GSK-3359609 comprises: CDR-H1, wherein CDR-H1 contains the amino acid sequence of DYAMH (SEQ ID NO: 1525); CDR-H2, wherein CDR-H2 contains the amino acid sequence of LISIYSDHTNYNQKFQG (SEQ ID NO: 1526); CDR-H3, wherein CDR-H3 contains the amino acid sequence of NNYGNYGWYFDV (SEQ ID NO: 1527); CDR-L1, wherein CDR-L1 contains the amino acid sequence of SASSVSYMH (SEQ ID NO: 1528); CDR-L2, wherein CDR-L2 contains the amino acid sequence of DTSKLAS (SEQ ID NO: 1529); and CDR-L3, wherein CDR-L3 contains the amino acid sequence of FQGSGYPYT (SEQ ID NO: 1530). In some embodiments, GSK-3359609 comprises VH and VL, wherein VH comprises the amino acid sequence of SEQ ID NO: 1531 and VL comprises the amino acid sequence of SEQ ID NO: 1532.

[0830] In some embodiments, the anti-ICOS antibody or its antigen-binding fragment comprises one or more CDR sequences of JTX-2011 having at most two conserved amino acid substitutions. In some embodiments, the anti-ICOS antibody or its antigen-binding fragment comprises VH and / or VL, said VH and / or said VL having an amino acid sequence that is at least 80% identical (e.g., 85%, 90%, 95%, 97%, 99%, or 100% identical) to the VH and / or VL sequence of JTX-2011. JTX-2011 and other anti-ICOS antibodies are disclosed, for example, in WO 2016 / 154177; which is incorporated herein by reference.

[0831] In some embodiments, JTX-2011 comprises: CDR-H1, wherein CDR-H1 contains the amino acid sequence of GFTFSDYWMD (SEQ ID NO: 1533); CDR-H2, wherein CDR-H2 contains the amino acid sequence of NIDEDGSITEYSPFVKG (SEQ ID NO: 1534); CDR-H3, wherein CDR-H3 contains the amino acid sequence of WGRFGFDS (SEQ ID NO: 1535); CDR-L1, wherein CDR-L1 contains the amino acid sequence of KSSQSLLSGSFNYLT (SEQ ID NO: 1536); CDR-L2, wherein CDR-L2 contains the amino acid sequence of YASTRHT (SEQ ID NO: 1537); and CDR-L3, wherein CDR-L3 contains the amino acid sequence of HHHYNAPPT (SEQ ID NO: 1538). In some embodiments, JTX-2011 comprises VH and VL, wherein VH comprises the amino acid sequence of SEQ ID NO: 1539 and VL comprises the amino acid sequence of SEQ ID NO: 1540.

[0832] Amino acid modifications at the Fc domain

[0833] The anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody, or their antigen-binding fragments, disclosed herein contain at least 50 amino acid residues of an Fc domain. The anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody, or their antigen-binding fragments, of the present disclosure, further contain one or more amino acid modifications (e.g., substitution, deletion, insertion, or chemical modification) at the Fc domain. Methods for introducing such amino acid modifications into peptides (e.g., the anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody, or their antigen-binding fragments, or their Fc domains, of the present disclosure) are known in the art. Those skilled in the art also understand that amino acid modifications at the Fc domain generally do not affect the antigen-binding properties (e.g., specificity, affinity, or cohesion) of the antibody or its antigen-binding fragment.

[0834] Human IgG Fc domain

[0835] Humans have four IgG subtypes, referred to as IgG1, IgG2, IgG3, and IgG4. Each of these subtypes corresponds to a different heavy chain constant region containing an Fc domain. The amino acid and DNA sequences of these heavy chain constant regions are known in the art and can be accessed using the UniProt database. Exemplary amino acid sequences of the human IgG1 heavy chain constant region can be accessed using UniProt accession number P01857. Exemplary amino acid sequences of the human IgG2 heavy chain constant region can be accessed using UniProt accession number P01859. Exemplary amino acid sequences of the human IgG3 heavy chain constant region can be accessed using UniProt accession number P01860. Exemplary amino acid sequences of the human IgG4 heavy chain constant region can be accessed using UniProt accession number P01861. The anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody, or their antigen-binding fragments, disclosed herein may have an IgG1 subtype, an IgG2 subtype, an IgG3 subtype, or an IgG4 subtype. In some embodiments, the anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody, or their antigen-binding fragments, of this disclosure have an IgG4 subtype. In some embodiments, the anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody, or their antigen-binding fragments, of this disclosure are IgG4 antibodies or their antigen-binding fragments.

[0836] The Fc domain of human IgG comprises a CH2 domain and a CH3 domain. The amino acid sequences of the CH2 and CH3 domains of human IgG are relatively conserved across different IgG isotypes. Table 15 below shows exemplary amino acid residues at each position of the CH2 domain of human IgG, and Table 16 below shows exemplary amino acid residues at each position of the CH3 domain of human IgG. Isotypes of these sequences exist that are known to those skilled in the art, with one or more amino acid residue modifications (e.g., substitution, deletion, or insertion). Unless otherwise stated, throughout this disclosure, the amino acid positions at the Fc domain are numbered according to the EU index.

[0837] Table 15. Amino acid residues at each position of the CH2 domain of human IgG.

[0838]

[0839]

[0840]

[0841]

[0842] Table 16. Amino acid residues at each position of the CH3 domain of human IgG.

[0843]

[0844]

[0845]

[0846] Human IgG hinge region

[0847] The CH1 and CH2 domains of human IgG are connected to the hinge region. The amino acid sequence of the hinge region of human IgG is known in the art and described, for example, in Kabat et al., Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md. 1987), the disclosure of which is incorporated herein by reference. Table 17 below shows exemplary amino acid residues at each position in the hinge region of human IgG. Isotypes of these sequences are known to those skilled in the art, with one or more amino acid residue modifications (e.g., substitution, deletion, or insertion). Unless otherwise stated, throughout this disclosure, the amino acid positions at the Fc domain are numbered according to the EU index.

[0848] Table 17. Amino acid residues at each location in the hinge region of human IgG.

[0849]

[0850] Reduce or eliminate amino acid modifications that reduce or eliminate Fc function

[0851] For a long time, it has been believed that functional antibodies (e.g., agonist or antagonist antibodies, such as those specifically binding to TNFRSF members, TNFSF members, CD28, or ICOS) are more potent when they possess a functional Fc receptor region. This is because the Fc region can bind to adjacent cells, thereby promoting the recruitment of effector cells. This antibody-bridged intercellular crosslinking is commonly referred to as ADCC (antibody-dependent cytotoxicity). This intercellular crosslinking increases the signal strength of the antibody across antigen-binding regions opposite to the Fc region on two different cells. Furthermore, functional antibodies typically bind to a single antigen via two antigen-binding arms. Antibody engineering is often performed to enhance the function of the antibody's Fc region, as this is generally expected to increase intercellular crosslinking and thus enhance antibody potency. Known Fc mutations exist for improving effector function, as shown in Table 18 below. It has long been believed that:

[0852] (a) Strong Fc receptor function is crucial for antibody function;

[0853] (b) Crosslinks mediated by antibodies modified to improve the function of Fc receptor effectors are located between two cells; and

[0854] (c) Antibodies (such as those modified to improve the function of Fc receptor effectors) typically bind to a single antigen via two antigen-binding arms.

[0855] Contrary to conventional thinking regarding the function of Fc receptor effectors of antibodies, the inventors have discovered a different method for enhancing the function of antibodies and their antigen-binding fragments. The unique feature of the functional antibodies and their antigen-binding fragments (e.g., agonist or antagonist antibodies, such as those specifically binding to TNFRSF members, TNFSF members, CD28, or ICOS) produced by the method described herein is that removing the Fc receptor region from the entire antibody (or modifying it to reduce, block, or eliminate Fc effector function) does not have a significant effect on altering antigen binding and does not impair the agonist or antagonist function of the antibody or its antigen-binding fragment. The receptor profiles of these antibodies show that they bind consistently to two adjacent antigens on the same cell. These antibodies also exhibit cell-surface-only crosslinking via what is considered a hexagonal network, where both the antibody and antigen are located entirely on the cell surface. The antibodies or their antigen-binding fragments produced using the inventors' method are characterized by:

[0856] (a) Removing the Fc region of an antibody (or reducing, blocking or eliminating changes in the function of Fc region effectors) does not impair the agonist or antagonist function of the antibody or its antigen-binding fragments;

[0857] (b) The antibody or its antigen-binding fragment cross-links at the cell surface, thereby pulling two adjacent receptors together; and

[0858] (c) The antibody or its antigen-binding fragments exhibit single-cell cross-linking only at the cell surface.

[0859] Therefore, the inventors attempted to create potent functional antibodies by testing the following assumptions: cell surface cross-linked antibodies with enhanced ADCC function would be the most potent, and cell surface cross-linked antibodies with degraded ADCC function would be weaker. The results obtained by the inventors and disclosed herein are surprising and contradict results known in the art for many years. When the inventors removed Fc receptor function from antibodies that only cross-linked the cell surface, they created antibodies that were highly potent not only at very low doses but also over a very wide dose range (e.g., the antibodies were functional at both very low and very high doses).

[0860] ADCC-crosslinked antibodies and cell surface-crosslinked antibodies are characterized (and are generally limited by a two-model dose-response curve) by producing a narrow dose range when used to treat patients. When the inventors retain the cell surface crosslinking and remove the ADCC function of the antibody, the resulting antibody no longer exhibits a two-model dose-response curve. Surprisingly, this observation suggests that retained or enhanced Fc receptor function is detrimental to antibody function. Therefore, this disclosure teaches that an extended dose-response curve for functional antibodies is best achieved by performing cell surface crosslinking on antibodies that inactivate only the Fc receptor.

[0861] Compared to corresponding antibodies without such amino acid modifications, antibodies of this disclosure (e.g., the anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody or their antigen-binding fragments) having amino acid modifications (e.g., substitutions) that reduce or eliminate (e.g., inactivate) Fc function can be administered at higher doses, at higher dosing frequencies, or over a wider dose range. In some embodiments, the anti-TNFRSF antibody, anti-TNFSF antibody, anti-CD28 antibody, or anti-ICOS antibody or their antigen-binding fragments may contain one or more amino acid modifications (e.g., substitutions, deletions, insertions, or chemical modifications) at the Fc domain that reduce or eliminate (e.g., inactivate) the function of the Fc domain. For example, amino acid modifications may reduce or eliminate (e.g., inactivate) the binding affinity of the antibody or its antigen-binding fragment to an Fc receptor (e.g., FcγRI, FcγRII, or FcγRIII) and / or the effector function of the antibody or its antigen-binding fragment. The effector function may be an effector function mediated by an Fc receptor (such as ADCC or ADCP). In some implementations, the amino acid modification may be located within the hinge region of the antibody or its antigen-binding fragment.

[0862] In some embodiments, the Fc domain is the human IgG1 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0863] (i) L235E;

[0864] (ii) L234A and L235A;

[0865] (iii) L234A, L235A and P329G;

[0866] (iv) L234E, L235F and P331S;

[0867] (v)D265A;

[0868] (vi) G237A;

[0869] (vii) E318A;

[0870] (viii) E233P;

[0871] (ix) G236R and L328R;

[0872] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0873] (xi) N297G or N297Q,

[0874] The amino acid positions are based on the EU index number.

[0875] In some embodiments, the Fc domain is the human IgG2 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0876] (i) D265A;

[0877] (ii) G237A;

[0878] (iii) E318A;

[0879] (iv) H268Q, V309L, A330S and P331S;

[0880] (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and

[0881] (vi) N297G or N297Q,

[0882] The amino acid positions are based on the EU index number.

[0883] In some embodiments, the Fc domain is the human IgG3 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0884] (i) L235E;

[0885] (ii) L234A and L235A;

[0886] (iii) L234A, L235A and P329G;

[0887] (iv) L234E, L235F and P331S;

[0888] (v)D265A;

[0889] (vi) G237A;

[0890] (vii) E318A;

[0891] (viii) E233P;

[0892] (ix) G236R and L328R;

[0893] (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and

[0894] (xi) N297G or N297Q,

[0895] The amino acid positions are based on the EU index number.

[0896] In some embodiments, the Fc domain is the human IgG4 Fc domain. In some embodiments, the one or more amino acid modifications are substitutions of one or more amino acids selected from the group consisting of:

[0897] (i) L235E;

[0898] (ii) D265A;

[0899] (iii) G237A;

[0900] (iv) E318A;

[0901] (v)E233P;

[0902] (vi) G236R and L328R; and

[0903] (vii) N297G or N297Q,

[0904] The amino acid positions are based on the EU index number.

[0905] In some embodiments, the antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P (according to Kabat's S241P). In some embodiments, the one or more amino acid modifications are one or more amino acid substitutions selected from the group consisting of:

[0906] (i) L235E; and

[0907] (ii) F234A and L235A,

[0908] The amino acid positions are based on the EU index number.

[0909] In some embodiments, the antibody or its antigen-binding fragment is an IgG4 antibody or its antigen-binding fragment. In some embodiments, the modified antibody or its antigen-binding fragment contains an amino acid substitution for S228P.

[0910] In some embodiments, the one or more amino acid modifications are the deletion of one or more amino acids. In some embodiments, the one or more amino acid deletions are the deletion of all amino acid residues of the Fc domain. In some embodiments, the antibody or its antigen-binding fragment lacks the Fc domain. The absence of the Fc domain results in the loss of ADCC-mediated effector function, which depends on the Fc domain. However, effector function resulting from cell surface crosslinking of the antibody or its antigen-binding fragment is unaffected and is the basis for the enhanced function of the antibody or its antigen-binding fragment lacking the Fc domain or Fc effector function described herein. The resulting antibody or its antigen-binding fragment binds to two adjacent antigens, and this function does not involve linking two adjacent cells via mechanisms such as ADCC. In some embodiments, the antibody or its antigen-binding fragment is selected from the group consisting of: single-chain Fv molecules (scFv), bifunctional antibodies, trifunctional antibodies, antibody-like protein scaffolds, Fv fragments, Fab fragments, F(ab')2 molecules, and tandem scFv (taFv). Those skilled in the art will expect that antibodies lacking the Fc domain or their antigen-binding fragments (such as those described herein) retain antigen-binding activity because the binding domain of the antibody or antigen-binding fragment remains intact.

[0911] Additional amino acid modifications that reduce or eliminate (e.g., inactivate) Fc function are known in the art and can be found, for example, in Saunders. Front. Immunol. 10:1296, 2019; incorporated herein by reference.

[0912] In some embodiments, according to the EU index, the modified human IgG4 Fc domain (including the IgG4 Fc hinge region) contains a proline (P) amino acid substitution for serine (S) at amino acid position 228. In some embodiments, the modified human IgG4 Fc domain contains an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 1547. The modified human IgG4 Fc domain may, for example, contain the amino acid sequence of SEQ ID NO: 1547.

[0913] In some implementations, the modified human IgG4 Fc domain is an IgG4*01 allotype, an IgG4*02 allotype, an IgG4*03 allotype, an IgG4*05 allotype, or an IgG4*06 allotype.

[0914] In some embodiments, the modified human IgG4 Fc domain includes a CH2 domain according to the human IgG4 amino acid sequence provided in Table 15. In some embodiments, the modified human IgG4 Fc domain includes a CH3 region according to the human IgG4 amino acid sequence provided in Table 16. In some embodiments, the modified human IgG4 Fc domain includes a hinge region according to the human IgG4 amino acid sequence provided in Table 17. In some embodiments, according to the EU index, the modified human IgG4 Fc domain includes a hinge region according to the human IgG4 amino acid sequence provided in Table 17, differing in that the proline (P) at amino acid position 228 is substituted for the serine (S) amino acid. In some embodiments, the modified human IgG4 Fc domain results in little or no effector function, i.e., little or no antibody-dependent cytotoxicity (ADCC) or antibody-dependent phagocytosis (ADCP).

[0915] IgG4 antibodies inherently possess fewer effector functions than IgG1 or IgG2 antibodies. For example, the effector function can be termed ADCC. IgG4 antibodies naturally possess many desirable characteristics, and other allotype antibodies can be modified to possess these characteristics. Natural IgG4 or other allotype antibodies can be modified to have less Fc receptor function to prevent ADCC or modified to have a lower ability to bind complement, thereby killing target cells. Furthermore, antibodies can be modified to remove the Fc region, resulting in a Fab'2 fragment that still has two arms but no longer possesses any effector function. As outlined herein, mutations in the hinges of antibodies can cause varying distances between antibody arms, which can affect the stability of crosslinks to achieve optimal agonistic effects.

[0916] The modified IgG4 antibodies disclosed herein possess the following advantageous properties. In some embodiments, the modified IgG4 antibodies exhibit enhanced agonistic activity. This enhanced agonistic activity indicates that the modified IgG4 antibodies are less susceptible to inhibition at higher doses compared to corresponding antibodies (e.g., IgG1 or IgG2 isotypes or IgG4 isotypes with one or more C mutations that enhance ADCC) having a greater effector function (e.g., ADCC). In some embodiments, the modified human IgG4 Fc domain on the modified IgG4 antibody reduces or inactivates the effector function of the modified IgG4 antibody compared to a control antibody (e.g., a modified human IgG1 or IgG2 antibody containing an IgG1 or IgG2 Fc domain or an unmodified IgG4 antibody containing an IgG4 Fc domain). For example, the effector function may be selected from antibody-dependent cytotoxicity (ADCC) or antibody-dependent phagocytosis (ADCP). In some embodiments, the modified human IgG4 Fc domain of the modified IgG4 antibody reduces complement binding function compared to a control antibody (e.g., a modified human IgG1 or IgG2 antibody containing an IgG1 or IgG2 Fc domain or an unmodified IgG4 antibody containing an IgG4 Fc domain). In some embodiments, the modified IgG4 antibody can be administered to the subject at a higher dose range and / or frequency than the control antibody (e.g., a modified human IgG1 or IgG2 antibody containing an IgG1 or IgG2 Fc domain or an unmodified IgG4 antibody containing an IgG4 Fc domain).

[0917] In some embodiments, compared to the corresponding antibody having unmodified IgG4 Fc, the modified IgG4 antibody contains one or more Fc substitutions that further reduce or eliminate effector functions (e.g., ADCC or ADCP).

[0918] In some implementations, the modified IgG4 antibody can maintain or enhance efficacy when administered to subjects at a higher dose range than the control antibody (e.g., a modified human IgG1 or IgG2 antibody containing an IgG1 or IgG2 Fc domain). For example, maintaining or enhancing efficacy at a higher dose may refer to maintaining or increasing Treg cell proliferation when administered to a subject or patient in need. Maintaining or enhancing efficacy at higher doses can also be monitored by measuring an increase in soluble TNFR2 (sTNFR2) in the blood (e.g., an increase of 1, 2, 3, or 4 times). sTNFR2 will optimally be increased to a level at or above 2 ng / ml, and at most 6 ng / ml. For example, the efficacy of the modified IgG4 antibody can be maintained or enhanced by subcutaneous (SQ) administration. The advantage of SQ administration compared to intravenous (IV) administration is that SQ administration allows for the proper formation of the modif...

Claims

1. A method for modulating the immune response of a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a member protein of the human tumor necrosis factor receptor superfamily (TNFRSF), wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof.

2. A method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a human TNFRSF member protein, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a more modulated dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency to produce the immune response.

3. The method of claim 2, wherein the modified antibody or its antigen-binding fragment is administered to the human subject at a higher dose and / or frequency than the unmodified antibody or its antigen-binding fragment.

4. The method according to any one of claims 1 to 3, wherein the TNFRSF member proteins are selected from the group consisting of: TNFR2, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TNFR1, TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3.

5. The method according to any one of claims 1 to 4, wherein the method further comprises: (a) Measure the pretreatment level of soluble TNFRSF member proteins in the serum or plasma of the human subject prior to administration of one or more doses of the modified antibody or its antigen-binding fragment; (b) Measuring the post-treatment levels of soluble TNFRSF member proteins in the serum or plasma of the human subject after administration of one or more doses of the modified antibody or its antigen-binding fragment; and / or (c) Determine the reference levels of soluble TNFRSF member proteins in the serum or plasma of healthy individuals.

6. The method according to claim 5, wherein: (a) The modified antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein, wherein: (i) The pre-treatment level of the soluble TNFRSF member protein in the human subject is lower than or the same as the reference level of the soluble TNFRSF member protein; and / or (ii) The post-treatment level of the soluble TNFRSF member protein in the human subject is higher than the reference level or the pre-treatment level of the soluble TNFRSF member protein, and the method further comprises administering one or more additional doses of the agonist antibody or its antigen-binding fragment to the human subject during one or more additional treatment periods to maintain the higher level; or (b) The modified antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein, wherein: (i) The pre-treatment level of the soluble TNFRSF member protein in the human subject was higher than the reference level of the soluble TNFRSF member protein; and / or (ii) The post-treatment level of the soluble TNFRSF member protein in the human subject is lower than the reference level or the pre-treatment level of the soluble TNFRSF member protein, and the method further comprises administering one or more additional doses of the antagonistic antibody or its antigen-binding fragment to the human subject during one or more additional treatment periods to maintain the lower level.

7. The method according to any one of claims 1 to 6, wherein the modified antibody or its antigen-binding fragment is an agonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

8. The method of claim 7, wherein the TNFRSF member protein is TNFR2.

9. The method of claim 8, wherein the modified antibody or its antigen-binding fragment comprises: (a) Heavy chain complementarity determining region (CDR) 1 (CDR-H1), wherein CDR-H1 comprises the following amino acid sequences: (i)X 1 X 2 X 2 X 1 X 2 X 3 X 2 JJJ, where: Each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; Each X 2 Independently, it is Y, S, T, C, N, or Q; X 3 It is either D or E; and Each J is independently a naturally occurring amino acid or is absent; or (ii)X 1 X 2 X 2 X 3 X 1 ,in: Each X 1 It can be either D or E independently; Each X 2 Independently, it is Y, S, T, C, N, or Q; and X 3 It is L, A, V, G, I, M, W, F, or P; (b) CDR-H2, wherein CDR-H2 contains X 1 X 2 X 3 X 2 X 3 X 3 X 1 X 3 X 3 X 3 X 3 X 3 X 3 X 4 X 2 X 4 X 2 The amino acid sequence, in which: Each X 1 It can be either D or E independently; Each X 2 Independently, it is I, A, V, L, G, M, W, F, or P; Each X 3 Independently, it is N, S, T, C, Y, or Q; and Each X 4 It can be K, R, or H independently; (c) CDR-H3, wherein CDR-H3 contains X 1 X 2 X 2 X 1 X 2 X 1 X 3 X 1 The amino acid sequence, in which: Each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; Each X 2 Independently, it is N, S, T, C, Y, or Q; and X 3 It is either D or E; (d) Light chain CDR 1 (CDR-L1), wherein CDR-L1 contains X 1 X 2 X 1 X 1 X 1 X 2 X 3 X 4 X 1 X 1 The amino acid sequence, in which: Each X 1 Independently, it is S, Y, T, C, N, or Q; Each X 2 Independently, it is A, G, V, L, I, M, W, F, or P; X 3 It is R, H, or K; and X 4 It is Y, A, V, I, L, M, F, or W; (e) CDR-L2, wherein the CDR-L2 includes X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, in which: Each X 1 Independently, it is L, A, V, G, I, M, W, F, or P; and Each X 2 Independently, it is T, S, C, Y, N, or Q; and / or (f) CDR-L3, wherein the CDR-L3 includes X 1 X 2 X 2 X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, in which: Each X 1 Independently, it is P, A, V, L, I, M, W, F, or G; and Each X 2 It can be Q, S, T, C, N, or Y independently.

10. The method of claim 9, wherein the CDR-H1 comprises GYTFTZ 1 Z 2 The amino acid sequence of JJJ (SEQ ID NO: 1480), wherein: Z 1 It is D or T; Z 2 It is Y, F, or L; and Each J is either a naturally occurring amino acid or not present.

11. The method of claim 10, wherein the CDR-H1 comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413).

12. The method according to any one of claims 9 to 11, wherein the modified antibody or its antigen-binding fragment further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 shown in Table 2.

13. The method of claim 8, wherein the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as shown in Table 2.

14. The method of claim 8, wherein the modified antibody or its antigen-binding fragment comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH and the VL have at least 80% identical amino acid sequences to those shown in Table 3.

15. The method of claim 7, wherein the TNFRSF member protein is selected from the group consisting of CD27, CD40, GITR, OX40 and 4-1BB.

16. The method of claim 15, wherein the modified antibody or its antigen-binding fragment comprises the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of an antibody selected from the group consisting of: varlilumab (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Tavolimab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utomilumab (PF-05082566), and Urelumab (BMS-663513).

17. The method of claim 15, wherein the modified antibody or its antigen-binding fragment comprises VH and VL, said VH and said VL having an amino acid sequence that is at least 80% identical to the VH and VL sequences of an antibody selected from the group consisting of: varirubin (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Talixicizumab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utoluumab (PF-05082566), and Ureluumab (BMS-663513).

18. The method according to any one of claims 7 to 17, wherein the serum or plasma level of the soluble TNFRSF member protein in the human subject is lower than the reference level of the soluble TNFRSF member protein.

19. The method according to any one of claims 7 to 18, wherein the modified antibody or its antigen-binding fragment exerts one or more biological activities selected from the group consisting of: (a) Promotes NFκB activation; (b) Inducing the proliferation of a population of T-reg cells; (c) Directly kills a population of CD8+ T cells or promotes the death of said population; (d) Promote the increase in the level of mRNA molecules encoding one or more proteins selected from the group consisting of: cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in the angiogenesis pathway, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α and lymphotoxin β; (e) Promotes increased levels of one or more proteins selected from the group consisting of: cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in the angiogenesis pathway, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α, and lymphotoxin β; (f) Promotes an increase in itaconic acid levels; and (g) Promotes the increase of the level of the soluble TNFRSF member protein.

20. The method according to any one of claims 1 to 6, wherein the modified antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

21. The method of claim 20, wherein the TNFRSF member protein is TNFR2.

22. The method of claim 21, wherein the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as shown in Table 4.

23. The method of claim 21, wherein the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and the VL have at least 80% identical amino acid sequences to those shown in Table 5.

24. The method according to any one of claims 20 to 23, wherein the serum or plasma level of the soluble TNFRSF member protein in the human subject is higher than a reference level of the soluble TNFRSF member protein.

25. The method according to any one of claims 20 to 24, wherein the modified antibody or its antigen-binding fragment exerts one or more biological activities selected from the group consisting of: (a) Inhibit NFκB activation; (b) Reduce or inhibit the proliferation of T-reg cell populations; (c) Inducing the proliferation of CD8+ T cell populations; (d) Reduce or inhibit the proliferation of cancer cell populations; (e) Inhibits TNFRSF member protein signaling in proliferating cells; (f) It does not inhibit TNFRSF member protein signaling in resting cells; (g) Reduce or inhibit the proliferation of myeloid-derived suppressor cells (MDSCs); (h) Promotes a decrease in the level of mRNA molecules encoding one or more proteins selected from the group consisting of: cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in the angiogenesis pathway, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α and lymphotoxin β; (i) Promotes a decrease in the levels of one or more proteins selected from the group consisting of: cIAP2, TRAF2, Etk, VEGFR2, PI3K, Akt, proteins involved in the angiogenesis pathway, IKK complex, RIP, NIK, MAP3K, proteins involved in the NFκB pathway, NIK, JNK, AP-1, MEK, MKK3, NEMO, IL2R, Foxp3, IL2, TNF, lymphotoxin α and lymphotoxin β; and (j) Promotes a decrease in the level of the soluble TNFRSF member proteins.

26. The method according to any one of claims 1 to 6, wherein the modified antibody or its antigen-binding fragment is neither an agonist nor an antagonist of the TNFRSF member protein.

27. The method of claim 26, wherein the TNFRSF member protein is TNFR2.

28. The method of claim 27, wherein the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as shown in Table 6.

29. The method of claim 27, wherein the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and the VL have at least 80% identical amino acid sequences to those shown in Table 7.

30. The method according to any one of claims 1 to 29, wherein the modified antibody or its antigen-binding fragment has less than about 50% cross-reactivity with the soluble TNFRSF member protein.

31. A method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a member protein of the human tumor necrosis factor superfamily (TNFSF), wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof.

32. A method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to a human TNFSF member protein, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a more modulated dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency that produces the immune response.

33. The method of claim 31 or 32, wherein the modified antibody or its antigen-binding fragment is administered to the human subject at a higher dose and / or frequency than the unmodified antibody or its antigen-binding fragment.

34. The method according to any one of claims 31 to 33, wherein the TNFSF member protein is TRAIL.

35. A method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human CD28, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof.

36. A method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human CD28, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a more modulated dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency that produces the immune response.

37. The method of claim 35 or 36, wherein the modified antibody or its antigen-binding fragment is administered to the human subject at a higher dose and / or frequency than the unmodified antibody or its antigen-binding fragment.

38. The method according to any one of claims 35 to 37, wherein the modified antibody or antigen-binding fragment thereof is an agonistic antibody or antigen-binding fragment thereof that specifically binds to CD28.

39. The method of claim 38, wherein the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of theralizumab (TAB-08).

40. The method of claim 38, wherein the modified antibody or its antigen-binding fragment comprises VH and VL, the VH and the VL having at least 80% identical amino acid sequences to the VH and VL sequences of cilalizumab (TAB-08).

41. A method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human ICOS, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a higher dose and / or frequency than an unmodified antibody or antigen-binding fragment thereof.

42. A method for modulating an immune response in a human subject, the method comprising administering to the subject a modified antibody or antigen-binding fragment thereof that specifically binds to human ICOS, wherein the modified antibody or antigen-binding fragment thereof comprises an Fc domain having one or more amino acid modifications, and wherein the modified antibody or antigen-binding fragment thereof is administered to the human subject at a more modulated dose and / or frequency relative to an unmodified antibody or antigen-binding fragment thereof administered at the stated dose and / or frequency that produces the immune response.

43. The method of claim 41 or 42, wherein the modified antibody or its antigen-binding fragment is administered to the human subject at a higher dose and / or frequency than the unmodified antibody or its antigen-binding fragment.

44. The method according to any one of claims 41 to 43, wherein the modified antibody or antigen-binding fragment thereof is an agonistic antibody or antigen-binding fragment thereof that specifically binds to ICOS.

45. The method of claim 44, wherein the modified antibody or its antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of GSK-3359609 or JTX-2011.

46. ​​The method of claim 44, wherein the modified antibody or its antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical to the VH and VL sequences of GSK-3359609 or JTX-2011.

47. The method according to any one of claims 1 to 46, wherein the one or more doses of the modified antibody or its antigen-binding fragment are administered to the human subject during one or more treatment periods, wherein: (a) Each dose contains: (i) about 0.1 mg to about 5000 mg of the modified antibody or its antigen-binding fragment; or (ii) the modified antibody or its antigen-binding fragment at a concentration of about 0.001 mg / kg to about 50 mg / kg; and / or (b) Each treatment period lasts for one, two, three, four, five, six, seven, eight or longer weeks.

48. The method according to any one of claims 1 to 47, wherein the modified antibody or antigen-binding fragment is administered to the human subject at a frequency of once or more per month, every three weeks, every two weeks, weekly, every six days, every five days, every four days, every three days, every two days, or daily.

49. The method according to any one of claims 1 to 48, wherein the modified antibody or its antigen-binding fragment is administered to the human subject in the following doses: (a) about 10 mg to about 5000 mg; or (b) Approximately 1 mg / kg to approximately 50 mg / kg.

50. The method according to any one of claims 1 to 49, wherein the one or more amino acid modifications comprise one or more substitutions, deletions, insertions, or chemical modifications of the Fc domain.

51. The method according to any one of claims 1 to 50, wherein the modified antibody or antigen-binding fragment thereof is selected from the group consisting of: monoclonal antibody or antigen-binding fragment thereof, polyclonal antibody or antigen-binding fragment thereof, human antibody or antigen-binding fragment thereof, humanized antibody or antigen-binding fragment thereof, primate-derived antibody or antigen-binding fragment thereof, bispecific antibody or antigen-binding fragment thereof, multispecific antibody or antigen-binding fragment thereof, bivariate immunoglobulin domain, monovalent antibody or antigen-binding fragment thereof, and chimeric antibody or antigen-binding fragment thereof.

52. The method of claim 51, wherein the modified antibody or its antigen-binding fragment is a human antibody, a humanized antibody, or a chimeric antibody or its antigen-binding fragment.

53. The method according to any one of claims 1 to 52, wherein: (a) The modification of one or more amino acids at the Fc domain reduces the binding affinity of the modified antibody or its antigen-binding fragment to the Fc receptor relative to the corresponding unmodified antibody or its antigen-binding fragment; (b) The effector function of the modified antibody or its antigen-binding fragment is independent of antibody-dependent cytotoxicity (ADCC) or antibody-dependent phagocytosis (ADCP). (c) The modification of one or more amino acids at the Fc domain enhances the bioactivity of the modified antibody or its antigen-binding fragment relative to the corresponding unmodified antibody or its antigen-binding fragment; (d) The modified antibody or its antigen-binding fragment exhibits dimer binding to its antigen at high doses, wherein the modified antibody or its antigen-binding fragment comprises a first arm and a second arm, wherein the first arm binds to the antigen, and wherein the second arm binds to an adjacent protein; and / or (e) The modified antibody or its antigen-binding fragment forms a hexagonal network with its antigen at high doses.

54. The method according to claim 53, wherein: (a) The Fc receptor is selected from the group consisting of: FcγRI, FcγRII and FcγRIII; (b) The bioactivity of the modified antibody or its antigen-binding fragment is independent of ADCC or ADCP; and / or (c) The adjacent protein is an antigen of the antibody or its antigen-binding fragment.

55. The method according to any one of claims 1 to 54, wherein the Fc domain is selected from the group consisting of: IgG Fc domain, IgA Fc domain, IgD Fc domain, IgE Fc domain and IgM Fc domain.

56. The method according to any one of claims 1 to 55, wherein the Fc domain is an IgG Fc domain.

57. The method according to any one of claims 1 to 56, wherein the Fc domain is the human IgG1 Fc domain.

58. The method of claim 57, wherein the one or more amino acid modifications are one or more amino acid substitutions selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q, Furthermore, the amino acid positions are based on EU index numbers.

59. The method according to any one of claims 1 to 56, wherein the Fc domain is the human IgG2 Fc domain.

60. The method of claim 59, wherein the one or more amino acid modifications are one or more amino acid substitutions selected from the group consisting of: (i) D265A; (ii) G237A; (iii) E318A; (iv) H268Q, V309L, A330S and P331S; (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and (vi) N297G or N297Q, Furthermore, the amino acid positions are based on EU index numbers.

61. The method according to any one of claims 1 to 56, wherein the Fc domain is the human IgG3 Fc domain.

62. The method of claim 61, wherein the one or more amino acid modifications are one or more amino acid substitutions selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q, Furthermore, the amino acid positions are based on EU index numbers.

63. The method according to any one of claims 1 to 56, wherein the Fc domain is the human IgG4 Fc domain.

64. The method of claim 63, wherein the one or more amino acid modifications are one or more amino acid substitutions selected from the group consisting of: (i) L235E; (ii) D265A; (iii) G237A; (iv) E318A; (v)E233P; (vi) G236R and L328R; and (vii) N297G or N297Q, Furthermore, the amino acid positions are based on EU index numbers.

65. The method of claim 63, wherein the modified antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P.

66. The method of claim 65, wherein the one or more amino acid modifications are one or more amino acid substitutions selected from the group consisting of: (i) L235E; and (ii) F234A and L235A, Furthermore, the amino acid positions are based on EU index numbers.

67. The method according to any one of claims 1 to 66, wherein the modified antibody or its antigen-binding fragment does not contain any of the amino acid substitutions shown in Table 18, and wherein the amino acid positions are according to EU index numbers.

68. The method according to any one of claims 1 to 67, wherein the modified antibody or its antigen-binding fragment is conjugated to a therapeutic agent.

69. The method of claim 68, wherein the therapeutic agent is selected from the group consisting of: chemotherapeutic agents, immunotherapeutic agents, agonists of TNFRSF member proteins, antagonists of TNFRSF member proteins, agonists of TNFSF member proteins, antagonists of TNFSF member proteins, agonists of CD28, antagonists of CD28, agonists of ICOS, and antagonists of ICOS.

70. The method according to any one of claims 1 to 69, wherein the method comprises administering an additional therapeutic agent.

71. The method of claim 70, wherein the additional therapeutic agent is selected from the group consisting of: chemotherapeutic agents, immunotherapeutic agents, agonists of TNFRSF member proteins, antagonists of TNFRSF member proteins, agonists of TNFSF member proteins, antagonists of TNFSF member proteins, agonists of CD28, antagonists of CD28, agonists of ICOS, and antagonists of ICOS.

72. The method according to any one of claims 1 to 71, wherein the modified antibody or its antigen-binding fragment is administered intravenously or subcutaneously to the human subject.

73. The method according to any one of claims 1 to 72, wherein the method inhibits the immune response mediated by B cells, CD8+ T cells or T-reg cells in the human subject.

74. The method according to any one of claims 1 to 72, wherein the method promotes an immune response mediated by B cells, CD8+ T cells or T-reg cells in the human subject.

75. The method according to any one of claims 1 to 74, wherein the human subject requires tissue or organ regeneration.

76. The method of claim 75, wherein the tissue or organ is selected from the group consisting of: pancreas, islets of Langerhans, salivary glands, pituitary gland, kidney, heart, lung, hematopoietic system, cranial nerves, heart, aorta, olfactory glands, ear, nerves, head structures, eye, thymus, tongue, bone, liver, small intestine, large intestine, intestine, lung, brain, skin, peripheral nervous system, central nervous system, spinal cord, mammary glands, embryonic structures, embryo, and testis.

77. The method according to any one of claims 1 to 76, wherein the method is used to treat cell proliferation disorders.

78. The method of claim 77, wherein the cell proliferation disorder is a cancer selected from the group consisting of: leukemia, lymphoma, liver cancer, bone cancer, lung cancer, brain cancer, bladder cancer, gastrointestinal cancer, breast cancer, heart cancer, cervical cancer, uterine cancer, head and neck cancer, gallbladder cancer, laryngeal cancer, lip and oral cancer, eye cancer, melanoma, pancreatic cancer, prostate cancer, colorectal cancer, testicular cancer, and pharyngeal cancer.

79. The method of claim 77, wherein the cell proliferation disorder is a cancer selected from the group consisting of: acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), adrenocortical carcinoma, AIDS-related lymphoma, primary central nervous system (CNS) lymphoma, anal cancer, appendiceal cancer, astrocytoma, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, extrahepatic cancer, Ewing sarcoma family, osteosarcoma and malignant fibrous histiocytoma, embryonal tumors of the central nervous system, germ cell tumors of the central nervous system, craniopharyngioma, ependymoma, bronchial tumor, Burkitt's lymphoma. Lymphoma, carcinoid tumor, primary lymphoma, chordoma, chronic myeloproliferative neoplasm, colon cancer, extrahepatic bile duct carcinoma, ductal carcinoma in situ (DCIS), endometrial cancer, ependymoma, esophageal cancer, olfactory neuroblastoma, extracranial germ cell tumor, gonadal germ cell tumor, fallopian tube cancer, osteofibrous histiocytoma, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), testicular germ cell tumor, gestational trophoblastic disease, glioma, childhood brainstem glioma, hairy cell leukemia, hepatocellular carcinoma, Langerhans cell histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, islet cell tumor, pancreatic neuroendocrine tumor, Wilms' tumor and other childhood kidney tumors, Langerhans cell histiocytosis, small cell lung cancer, cutaneous T-cell lymphoma, intraocular melanoma, Merkel cell carcinoma. Carcinoma, mesothelioma, metastatic squamous neck carcinoma, midline carcinoma, multiple endocrine adenoma syndrome, multiple myeloma / plasma cell tumor (e.g., multiple myeloma or refractory multiple myeloma), myelodysplastic syndrome, nasal cavity and sinus carcinoma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin lymphoma,NHL), non-small cell lung cancer (NSCLC), epithelial ovarian cancer, germ cell ovarian cancer, low-grade malignant potential ovarian cancer, pancreatic neuroendocrine tumors, papilloma, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid carcinoma, penile cancer, pharyngeal cancer, pheochromocytoma, pituitary adenoma, pleural pulmonary blastoma, primary peritoneal cancer, rectal cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, Kaposi sarcoma, rhabdomyosarcoma, Sézary syndrome, small bowel cancer, soft tissue sarcoma, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, urethral cancer, endometrial uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, and Waldenström macroglobulinemia.

80. The method according to any one of claims 77 to 79, wherein the modified antibody or its antigen-binding fragment is: (a) A stimulatory antibody or its antigen-binding fragment that specifically binds to TNFRSF member proteins selected from the group consisting of: TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TNFR1, Fas, CD40, CD27, 4-1BB, OX40, GITR, and XEDAR; or (b) Antagonistic antibodies or their antigen-binding fragments that specifically bind to TNFRSF member proteins selected from the group consisting of: TRAMP, NGFR, TRAIL-R4, TNFR2, HVEM, CD30, TROY, and RELT.

81. The method of claim 80, wherein the modified antibody or antigen-binding fragment thereof is an antagonistic antibody or antigen-binding fragment thereof that specifically binds to TNFR2 comprising: CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as shown in Table 4.

82. The method of claim 80, wherein the modified antibody or its antigen-binding fragment is an antagonistic antibody or its antigen-binding fragment that specifically binds to TNFR2 comprising VH and VL, wherein the VH and the VL have an amino acid sequence that is at least 80% identical to the sequences shown in Table 5.

83. The method according to any one of claims 1 to 76, wherein the method is used to treat an autoimmune disease or inflammatory disease in the human subject.

84. The method of claim 83, wherein the autoimmune disease is selected from the group consisting of: type 1 diabetes mellitus, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, Behçet's disease, bullous pemphigoid, cardiomyopathy, celiac dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, localized scleroderma (CREST syndrome), cold agglutinin disease, Crohn's disease, discoid lupus erythematosus, primary mixed cryoglobulinemia vasculitis, fibromyalgia-fibromyalgia, Graves' disease, Guillain-Barré syndrome. Hashimoto's thyroiditis, hypothyroidism, inflammatory bowel disease, autoimmune lymphoproliferative syndrome (ALPS), idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, juvenile arthritis, lichen planus, lupus, lupus nephritis, Ménière's disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychondritis, polygonococcal syndrome, polymyositis rheumatica, polymyositis, dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome Stiff-Man syndrome, Takayasu's arteritis, temporal arteritis / giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, Wegener's granulomatosis, Goodpasture syndrome, transplant rejection, celiac disease, esophagitis, and inflammatory CNS conditions, optionally including Alzheimer's disease.

85. The method according to any one of claims 1 to 76, wherein the method is used to treat an infectious disease in the human subject.

86. The method of claim 85, wherein the infectious disease is caused by one or more infectious agents selected from the group consisting of viruses, bacteria, fungi, and parasites.

87. The method of claim 86, wherein the infectious disease is caused by a virus selected from the group consisting of: hepatitis C virus, yellow fever virus, Kadam virus, Kyasanur forest disease virus, Langat virus, Omsk hemorrhagic fever virus, Powassan virus, Royal Farm virus, Karshi virus, tick-borne encephalitis virus, Neudoerfl virus, Sofjin virus, sheep jumping disease virus, Negishi virus, Meaban virus, Saumarez Reef virus, Tyuleniy virus, Aroa virus, dengue virus, Kedougou virus, Cacipacore virus, Koutango virus. Virus, Japanese encephalitis virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Usutu virus, West Nile virus, Yaounde virus, Kokobera virus, Bagaza virus, Ilheus virus, Israel turkeymeningoencephalomyelitis virus, Ntaya virus, Tembusu virus, Zika virus, Banzi virus, Bouboui virus, Edge Hill virus, Jugra virus, Saboya virus, Sepik virus, Uganda S virus Wesselsbron virus, yellow fever virus, Entebbe bat virusbat virus, Yokose virus, Apoi virus, Cowbone Ridge virus, Jutiapa virus, Modoc virus, Sal Vieja virus, San Perlita virus, Bukalasa bat virus, Carey Island virus, Dakar bat virus, Montana myotis leukoencephalitis virus, Phnom Penh bat virus, Rio Bravo virus, Tamana bat virus, cell fusion agent virus, Ippy virus, Lassa virus, Lymphocytic choroid plexus meningitis virus (LCMV), Mobala virus, Mopeia virus virus, Amaparivirus, Flexal virus, Guanarito virus, Junin virus, Latino virus, Machupo virus, Oliveros virus, Paraná virus, Pichinde virus, Pirital virus, Sabiá virus, Tacaribe virus, Tamiami virus, Whitewater Arroyo virus, Chapare virus, Lujo virus, Hantaan virus, SinNombre virus, Dugbe virus, Bunyamwera virus, Rift Valley fever virus, La Crosse virus, California encephalitisencephalitis virus, Crimean-Congo hemorrhagic fever (CCHF) virus, Ebola virus, Marburg virus, Venezuelan equine encephalitis virus (VEE), Eastern equine encephalitis virus (EEE), Western equine encephalitis virus (WEE), Sindbis virus, rubella virus, Semliki Forest virus, Ross River virus, Barmah Forest virus, O'nyong'nyong virus, chikungunya virus, smallpox virus, monkeypox virus, vaccinia virus, herpes simplex virus, human herpesvirus, cytomegalovirus (CMV), Epstein-Barr virus. Human varicella-zoster virus (EBV), Kaposi's sarcoma-associated herpesvirus (KSHV), influenza virus, severe acute respiratory syndrome (SARS) virus, rabies virus, vesicular stomatitis virus (VSV), human respiratory syncytial virus (RSV), Newcastle disease virus, hendra virus, nipah virus, measles virus, rinderpest virus, canine distemper virus, Sendai virus, human parainfluenza virus (e.g., types 1, 2, 3, and 4), rhinovirus, mumps virus, poliovirus, human enterovirus (e.g., types A, B, C, and D), hepatitis A virus, coxsackie virus, hepatitis B virus, human papillomavirus, adeno-associated virus, astrovirus, JC virus, BK virus, SV40 virus, Norwalk virus. The viruses include rotavirus, human immunodeficiency virus (HIV), and human T-lymphotropic virus types I and II.

88. The method of claim 86, wherein the infectious disease is caused by bacteria belonging to a genus selected from the group consisting of: Salmonella, Streptococcus, Bacillus, Listeria, Corynebacterium, Nocardia, Neisseria, Acinetobacter, Moraxella, Enterobacteriaceae, Pseudomonas, Escherichia, Klebsiella. a) Serratia, Enterobacter, Proteus, Salmonella, Shigella, Yersinia, Haemophilus, Bordetella, Legionella, Pasteurella, Francisella, Brucella, Bartonella, Clostridium, Vibrio, Campylobacter, and Staphylococcus.

89. The method of claim 86, wherein the infectious disease is caused by fungi belonging to genera selected from the group consisting of: Aspergillus, Candida, Malassezia, Trichosporon, Fusarium, Acremonium, Rhizopus, Mucor, Pneumocystis, and Absidia.

90. The method of claim 86, wherein the infectious disease is caused by a parasite selected from the group consisting of: Entamoeba histolytica, Giardia lamblia, Cryptosporidium muris, Trypanosomatida gambiense, Trypanosomatida rhodesiense, Trypanosomatida crusi, Leishmania mexicana, Leishmania braziliensis, Leishmania tropica, Leishmania donovani, Toxoplasma gondii, Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, and Plasmodium falciparum. The parasites include falciparum, Trichomonas vaginalis, and Histomonas meleagridis.Exemplary worm parasites include *Trichuris trichiura*, *Ascaris lumbricoides*, *Enterobius vermicularis*, *Ancylostoma duodenale*, *Necator americanus*, *Strongyloides stercoralis*, *Wuchereria bancrofti*, *Dracunculus medinensis*, *Schistosoma mansoni*, *Schistosoma haematobium*, *Schistosoma japonicum*, *Fasciola hepatica*, *Fasciolagigantica*, *Heterophyes*, *Paragonimus westermani*, and *Taenia solium*. The following are listed: solium, beef tapeworm (Taenia saginata), hymenolepis nana, and echinococccus granulosus.

91. The method according to any one of claims 1 to 76, wherein the method is used to treat an inflammatory disease in the human subject.

92. The method of claim 91, wherein the inflammatory disease is selected from the group consisting of: acute or chronic inflammation, cardiac fibrosis, pulmonary fibrosis, osteoarthritis, rheumatoid arthritis, atherosclerosis, type I diabetes, type II diabetes, graft-versus-host disease, multiple sclerosis, osteomyelitis, psoriasis, Crohn's disease, Sjögren's syndrome, lupus erythematosus, and ulcerative colitis.

93. The method according to any one of claims 1 to 76, wherein the method is used to treat a neurological disease or condition of the human subject.

94. The method of claim 93, wherein the neurological disease or condition is selected from the group consisting of: brain tumor, brain metastasis, brain injury, spinal cord injury, nerve injury, schizophrenia, epilepsy, Parkinson's disease, autism, Huntington's disease, stroke, Alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), and myasthenia gravis.

95. The method according to any one of claims 1 to 76, wherein the method is used to treat allergies in the human subject.

96. The method of claim 95, wherein the allergy is selected from the group consisting of: food allergies, seasonal allergies, pet allergies, urticaria, hay fever, allergic conjunctivitis, poison ivy allergy, oak allergy, mold allergy, drug allergy, dust allergy, cosmetic allergy, and chemical allergy.

97. The method according to any one of claims 1 to 76, wherein the method is used to treat transplant rejection in the human subject.

98. The method of claim 97, wherein the transplant rejection is selected from the group consisting of: skin graft rejection, bone graft rejection, vascular tissue graft rejection, ligament graft rejection and organ graft rejection.

99. The method of claim 98, wherein the ligament graft rejection is selected from the group consisting of: cricothyroid ligament graft rejection, periodontal ligament graft rejection, lens suspensory ligament graft rejection, radial carpal ligament graft rejection, radial carpal dorsal ligament graft rejection, ulnar collateral ligament graft rejection, radial collateral ligament graft rejection, breast suspensory ligament graft rejection, anterior sacroiliac ligament graft rejection, posterior sacroiliac ligament graft rejection, sacrotuberous ligament graft rejection, sacrospinous ligament graft rejection, infrapubic ligament graft rejection, suprapubic ligament graft rejection, anterior cruciate ligament graft rejection, lateral collateral ligament graft rejection, posterior cruciate ligament graft rejection, medial collateral ligament graft rejection, craniocarpal ligament graft rejection, caudal cruciate ligament graft rejection, and patellar ligament graft rejection.

100. The method of claim 98, wherein the organ graft rejection is selected from the group consisting of: heart graft rejection, lung graft rejection, kidney graft rejection, liver graft rejection, pancreas graft rejection, intestinal graft rejection and thymus graft rejection.

101. The method according to any one of claims 1 to 76, wherein the method is used to treat graft-versus-host disease in the human subject.

102. The method of claim 101, wherein the graft-versus-host disease is caused by bone marrow transplantation or by one or more blood cells selected from the group consisting of: hematopoietic stem cells, common myeloid progenitor cells, common lymphoid progenitor cells, megakaryocytes, monocytes, basophils, eosinophils, neutrophils, macrophages, T cells, B cells, natural killer cells, and dendritic cells.

103. The method according to any one of claims 1 to 76, wherein the method is used for transplantation or for regeneration of tissues or organs of the human subject.

104. The method of claim 103, wherein the tissue or organ is selected from the group consisting of: pancreas, islets of Langerhans, salivary glands, pituitary gland, kidney, heart, lung, hematopoietic system, cranial nerves, heart, aorta, olfactory glands, ear, nerve, eye, thymus, tongue, bone, liver, small intestine, large intestine, gastrointestinal tract, lung, brain, skin, peripheral nervous system, CNS, spinal cord, mammary glands, embryonic structures, embryo, and testis.

105. The method according to claim 103 or 104, wherein the tissue or organ expresses the TNFRSF member protein, the TNFSF member protein, CD28, or ICOS.

106. An antibody or antigen-binding fragment thereof that specifically binds to a member protein of human TNFRSF, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, wherein: (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) D265A; (ii) G237A; (iii) E318A; (iv) H268Q, V309L, A330S and P331S; (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and (vi) N297G or N297Q; (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) D265A; (iii) G237A; (iv) E318A; (v)E233P; (vi) G236R and L328R; and (vii) N297G or N297Q; or (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P, the Fc domain being a human IgG4 Fc domain, and the one or more amino acid substitutions being selected from the group consisting of: (i) L235E; and (ii) F234A and L235A, Furthermore, the amino acid positions are based on EU index numbers.

107. The antibody or antigen-binding fragment thereof according to claim 106, wherein the TNFRSF member protein is selected from the group consisting of: 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TNFR1, TNFR2, TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3.

108. The antibody or antigen-binding fragment thereof according to claim 106 or 107, wherein the antibody or antigen-binding fragment thereof is an agonistic antibody or antigen-binding fragment thereof that specifically binds to the TNFRSF member protein.

109. The antibody or antigen-binding fragment thereof according to claim 108, wherein the TNFRSF member protein is TNFR2.

110. The antibody or antigen-binding fragment thereof according to claim 109, wherein the antibody or antigen-binding fragment thereof comprises: (a) CDR-H1, wherein CDR-H1 comprises the following amino acid sequences: (i)X 1 X 2 X 2 X 1 X 2 X 3 X 2 JJJ, where: Each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; Each X 2 Independently, it is Y, S, T, C, N, or Q; X 3 It is either D or E; and Each J is independently a naturally occurring amino acid or is absent; or (ii)X 1 X 2 X 2 X 3 X 1 ,in: Each X 1 It can be either D or E independently; Each X 2 Independently, it is Y, S, T, C, N, or Q; and X 3 It is L, A, V, G, I, M, W, F, or P; (b) CDR-H2, wherein CDR-H2 contains X 1 X 2 X 3 X 2 X 3 X 3 X 1 X 3 X 3 X 3 X 3 X 3 X 3 X 4 X 2 X 4 X 2 The amino acid sequence, in which: Each X 1 It can be either D or E independently; Each X 2 Independently, it is I, A, V, L, G, M, W, F, or P; Each X 3 Independently, it is N, S, T, C, Y, or Q; and Each X 4 It can be K, R, or H independently; (c) CDR-H3, wherein CDR-H3 contains X 1 X 2 X 2 X 1 X 2 X 1 X 3 X 1 The amino acid sequence, in which: Each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; Each X 2 Independently, it is N, S, T, C, Y, or Q; and X 3 It is either D or E; (d) CDR-L1, wherein CDR-L1 includes X 1 X 2 X 1 X 1 X 1 X 2 X 3 X 4 X 1 X 1 The amino acid sequence, in which: Each X 1 Independently, it is S, Y, T, C, N, or Q; Each X 2 Independently, it is A, G, V, L, I, M, W, F, or P; X 3 It is R, H, or K; and X 4 It is Y, A, V, I, L, M, F, or W; (e) CDR-L2, wherein the CDR-L2 includes X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, in which: Each X 1 Independently, it is L, A, V, G, I, M, W, F, or P; and Each X 2 Independently, it is T, S, C, Y, N, or Q; and / or (f) CDR-L3, wherein the CDR-L3 includes X 1 X 2 X 2 X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, in which: Each X 1 Independently, it is P, A, V, L, I, M, W, F, or G; and Each X 2 It can be Q, S, T, C, N, or Y independently.

111. The antibody or antigen-binding fragment thereof according to claim 110, wherein the CDR-H1 comprises GYTFTZ. 1 Z 2 The amino acid sequence of JJJ (SEQ ID NO: 1480), wherein: Z 1 It is D or T; Z 2 It is Y, F, or L; and Each J is either a naturally occurring amino acid or not present.

112. The antibody or antigen-binding fragment thereof according to claim 111, wherein the CDR-H1 comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413).

113. The antibody or antigen-binding fragment thereof according to any one of claims 110 to 112, wherein the antibody or antigen-binding fragment thereof further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 shown in Table 2.

114. The antibody or antigen-binding fragment thereof according to claim 109, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 shown in Table 2.

115. The antibody or antigen-binding fragment thereof according to claim 109, wherein the antibody or antigen-binding fragment thereof comprises VH and VL, wherein the VH and the VL have an amino acid sequence that is at least 80% identical to the sequence shown in Table 3.

116. The antibody or antigen-binding fragment thereof according to claim 108, wherein the TNFRSF member protein is selected from the group consisting of: CD27, CD40, GITR, OX40 and 4-1BB.

117. The antibody or antigen-binding fragment thereof according to claim 116, wherein the antibody or antigen-binding fragment thereof comprises the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of an antibody selected from the group consisting of: varirubin (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Talixicizumab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utoluumab (PF-05082566), and Ureluumab (BMS-663513).

118. The antibody or antigen-binding fragment thereof according to claim 116, wherein the antibody or antigen-binding fragment comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical to the VH and VL sequences of an antibody selected from the group consisting of: variliterumab (CDX-1127), CDX-1140, SEA-CD40, RO7009789, JNJ-64457107 (ADC1013), APX-005M, Chi Lob 7 / 4, TRX-518, MK-4166, MK-1248, GWN-323, INCAGN01876, BMS-986156, AMG-228, Talixicizumab (MEDI0562), PF-04518600, BMS-986178, MOXR-0916, GSK-3174998, INCAGN01949, Utoluumab (PF-05082566), and Ureluumab (BMS-663513).

119. The antibody or antigen-binding fragment thereof according to claim 106 or 107, wherein the antibody or antigen-binding fragment thereof is an antagonistic antibody or antigen-binding fragment thereof that specifically binds to the TNFRSF member protein.

120. The antibody or antigen-binding fragment thereof according to claim 119, wherein the TNFRSF member protein is TNFR2.

121. The antibody or antigen-binding fragment thereof according to claim 120, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 shown in Table 4.

122. The antibody or antigen-binding fragment thereof according to claim 120, wherein the antibody or antigen-binding fragment thereof comprises VH and VL, wherein the VH and the VL have an amino acid sequence that is at least 80% identical to the sequence shown in Table 5.

123. The antibody or antigen-binding fragment thereof according to claim 106 or 107, wherein the antibody or antigen-binding fragment thereof is neither an agonist nor an antagonist of the TNFRSF member protein.

124. The antibody or antigen-binding fragment thereof according to claim 123, wherein the TNFRSF member protein is TNFR2.

125. The antibody or antigen-binding fragment thereof according to claim 124, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 shown in Table 6.

126. The antibody or antigen-binding fragment thereof according to claim 124, wherein the antibody or antigen-binding fragment thereof comprises VH and VL, wherein the VH and the VL have an amino acid sequence that is at least 80% identical to the sequences shown in Table 7.

127. The antibody or antigen-binding fragment thereof according to any one of claims 106 to 126, wherein the antibody or antigen-binding fragment thereof has less than about 50% cross-reactivity with the soluble TNFRSF member protein.

128. An agonist antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, said agonist antibody or antigen-binding fragment comprising: (A) A component for binding to an epitope in human TNFR2, said component comprising: (a) CDR-H1, wherein CDR-H1 comprises the following amino acid sequences: (i)X 1 X 2 X 2 X 1 X 2 X 3 X 2 JJJ, where: Each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; Each X 2 Independently, it is Y, S, T, C, N, or Q; X 3 It is either D or E; and Each J is independently a naturally occurring amino acid or is absent; or (ii)X 1 X 2 X 2 X 3 X 1 ,in: Each X 1 It can be either D or E independently; Each X 2 Independently, it is Y, S, T, C, N, or Q; and X 3 It is L, A, V, G, I, M, W, F, or P; (b) CDR-H2, wherein CDR-H2 contains X 1 X 2 X 3 X 2 X 3 X 3 X 1 X 3 X 3 X 3 X 3 X 3 X 3 X 4 X 2 X 4 X 2 The amino acid sequence, in which: Each X 1 It can be either D or E independently; Each X 2 Independently, it is I, A, V, L, G, M, W, F, or P; Each X 3 Independently, it is N, S, T, C, Y, or Q; and Each X 4 It can be K, R, or H independently; (c) CDR-H3, wherein CDR-H3 contains X 1 X 2 X 2 X 1 X 2 X 1 X 3 X 1 The amino acid sequence, in which: Each X 1 Independently, it is G, A, V, L, I, M, W, F, or P; Each X 2 Independently, it is N, S, T, C, Y, or Q; and X 3 It is either D or E; (d) Light chain CDR 1 (CDR-L1), wherein CDR-L1 contains X 1 X 2 X 1 X 1 X 1 X 2 X 3 X 4 X 1 X 1 The amino acid sequence, in which: Each X 1 Independently, it is S, Y, T, C, N, or Q; Each X 2 Independently, it is A, G, V, L, I, M, W, F, or P; X 3 It is R, H, or K; and X 4 It is Y, A, V, I, L, M, F, or W; (e) CDR-L2, wherein the CDR-L2 includes X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, in which: Each X 1 Independently, it is L, A, V, G, I, M, W, F, or P; and Each X 2 Independently, it is T, S, C, Y, N, or Q; and / or (f) CDR-L3, wherein the CDR-L3 includes X 1 X 2 X 2 X 1 X 2 X 2 X 2 X 1 X 1 X 2 The amino acid sequence, in which: Each X 1 Independently, it is P, A, V, L, I, M, W, F, or G; and Each X 2 Independently, it is Q, S, T, C, N, or Y; and (B) An Fc domain, wherein the Fc domain comprises one or more amino acid substitutions, wherein: (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) D265A; (ii) G237A; (iii) E318A; (iv) H268Q, V309L, A330S and P331S; (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and (vi) N297G or N297Q; (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) D265A; (iii) G237A; (iv) E318A; (v)E233P; (vi) G236R and L328R; and (vii) N297G or N297Q; or (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P, the Fc domain being a human IgG4 Fc domain, and the one or more amino acid substitutions being selected from the group consisting of: (i) L235E; and (ii) F234A and L235A, Furthermore, the amino acid positions are based on EU index numbers.

129. The antibody or antigen-binding fragment thereof according to claim 120, wherein the CDR-H1 comprises GYTFTZ. 1 Z 2 The amino acid sequence of JJJ (SEQ ID NO: 1480), wherein: Z 1 It is D or T; Z 2 It is Y, F, or L; and Each J is either a naturally occurring amino acid or not present.

130. The antibody or antigen-binding fragment thereof according to claim 129, wherein the CDR-H1 comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413).

131. The antibody or antigen-binding fragment thereof according to any one of claims 128 to 130, wherein the antibody or antigen-binding fragment thereof further comprises one or more of the following: CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 shown in Table 2.

132. An antibody or antigen-binding fragment thereof that specifically binds to a human TNFSF member protein, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, wherein: (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) D265A; (ii) G237A; (iii) E318A; (iv) H268Q, V309L, A330S and P331S; (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and (vi) N297G or N297Q; (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) D265A; (iii) G237A; (iv) E318A; (v)E233P; (vi) G236R and L328R; and (vii) N297G or N297Q; or (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P, the Fc domain being a human IgG4 Fc domain, and the one or more amino acid substitutions being selected from the group consisting of: (i) L235E; and (ii) F234A and L235A, Furthermore, the amino acid positions are based on EU index numbers.

133. The antibody or antigen-binding fragment thereof according to claim 132, wherein the TNFSF member protein is TRAIL.

134. An antibody or antigen-binding fragment thereof that specifically binds to human CD28, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, wherein: (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) D265A; (ii) G237A; (iii) E318A; (iv) H268Q, V309L, A330S and P331S; (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and (vi) N297G or N297Q; (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) D265A; (iii) G237A; (iv) E318A; (v)E233P; (vi) G236R and L328R; and (vii) N297G or N297Q; or (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P, the Fc domain being a human IgG4 Fc domain, and the one or more amino acid substitutions being selected from the group consisting of: (i) L235E; and (ii) F234A and L235A, Furthermore, the amino acid positions are based on EU index numbers.

135. The antibody or antigen-binding fragment thereof according to claim 134, wherein the antibody or antigen-binding fragment thereof is an agonistic antibody or antigen-binding fragment thereof that specifically binds to CD28.

136. The antibody or antigen-binding fragment thereof according to claim 135, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of cilalizumab (TAB-08).

137. The antibody or antigen-binding fragment thereof according to claim 135, wherein the antibody or antigen-binding fragment thereof comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical to the VH and VL sequences of cilalizumab (TAB-08).

138. An antibody or antigen-binding fragment thereof that specifically binds to human ICOS, wherein the antibody or antigen-binding fragment comprises at least 50 amino acid residues of an Fc domain and one or more amino acid substitutions within the Fc domain, wherein: (a) The Fc domain is the human IgG1 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (b) The Fc domain is the human IgG2 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) D265A; (ii) G237A; (iii) E318A; (iv) H268Q, V309L, A330S and P331S; (v) V234A, G237A, P238S, H268A, V309L, A330S, and P331S; and (vi) N297G or N297Q; (c) The Fc domain is the human IgG3 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) L234A and L235A; (iii) L234A, L235A and P329G; (iv) L234E, L235F and P331S; (v)D265A; (vi) G237A; (vii) E318A; (viii) E233P; (ix) G236R and L328R; (x) L234A, L235A, G237A, P238S, H268A, A330S, and P331S; and (xi) N297G or N297Q; (d) The Fc domain is the human IgG4 Fc domain, and the one or more amino acid substitutions are selected from the group consisting of: (i) L235E; (ii) D265A; (iii) G237A; (iv) E318A; (v)E233P; (vi) G236R and L328R; and (vii) N297G or N297Q; or (e) The antibody or its antigen-binding fragment comprises a human IgG4 hinge region having an amino acid substitution S228P, the Fc domain being a human IgG4 Fc domain, and the one or more amino acid substitutions being selected from the group consisting of: (i) L235E; and (ii) F234A and L235A, Furthermore, the amino acid positions are based on EU index numbers.

139. The antibody or antigen-binding fragment thereof according to claim 138, wherein the antibody or antigen-binding fragment thereof is an agonistic antibody or antigen-binding fragment thereof that specifically binds to ICOS.

140. The antibody or antigen-binding fragment thereof according to claim 139, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of GSK-3359609 or JTX-2011.

141. The antibody or antigen-binding fragment thereof according to claim 139, wherein the antibody or antigen-binding fragment thereof comprises VH and VL, wherein the VH and VL have an amino acid sequence that is at least 80% identical to the VH and VL sequences of GSK-3359609 or JTX-2011.

142. The antibody or antigen-binding fragment thereof according to any one of claims 106 to 141, wherein the antibody or antigen-binding fragment thereof does not contain any of the amino acid substitutions shown in Table 18, and wherein the amino acid positions are according to EU index numbers.

143. The antibody or antigen-binding fragment thereof according to any one of claims 106 to 142, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of: monoclonal antibodies or antigen-binding fragments thereof, polyclonal antibodies or antigen-binding fragments thereof, human antibodies or antigen-binding fragments thereof, humanized antibodies or antigen-binding fragments thereof, primate-derived antibodies or antigen-binding fragments thereof, bispecific antibodies or antigen-binding fragments thereof, multispecific antibodies or antigen-binding fragments thereof, bivariate immunoglobulin domains, monovalent antibodies or antigen-binding fragments thereof, and chimeric antibodies or antigen-binding fragments thereof.

144. The antibody or antigen-binding fragment thereof according to claim 143, wherein the antibody or antigen-binding fragment thereof is a human antibody, a humanized antibody or a chimeric antibody or an antigen-binding fragment thereof.

145. A method for producing an antibody or an antigen-binding fragment thereof according to any one of claims 106 to 144, the method comprising expressing a polynucleotide encoding the antibody or an antigen-binding fragment thereof in a host cell and recovering the antibody or an antigen-binding fragment thereof from a host cell culture medium.

146. A construct comprising a first polypeptide domain and a second polypeptide domain, wherein the first polypeptide domain and the second polypeptide domain are each independently an antigen-binding fragment according to any one of claims 106 to 144.

147. The construct of claim 146, wherein the first polypeptide domain and the second polypeptide domain are linked by a covalent linker.

148. The construct of claim 147, wherein the covalent joint comprises an amide bond or a disulfide bond.

149. A polynucleotide encoding an antibody or antigen-binding fragment thereof according to any one of claims 106 to 144, or a construct according to any one of claims 146 to 148.

150. A vector comprising the polynucleotide according to claim 149.

151. The vector according to claim 150, wherein the vector is an expression vector or a viral vector.

152. The carrier according to claim 151, wherein: (a) The expression vector is a eukaryotic expression vector; or (b) The viral vector is selected from the group consisting of: adenovirus, retrovirus, poxvirus, adeno-associated virus, baculovirus, herpes simplex virus and vaccinia virus.

153. The carrier according to claim 152, wherein: (a) The adenovirus is serotype 1-60 adenovirus; (b) The adenovirus is serotype 5, 26, 35 or 48 adenovirus; (c) The retrovirus is a gamma-retrovirus or a lentivirus; or (d) The vaccinia virus is a modified Ankara vaccinia (MVA).

154. An isolated host cell, said isolated host cell comprising a vector according to any one of claims 150 to 153.

155. The host cell according to claim 154, wherein the host cell is a prokaryotic cell or a eukaryotic cell.

156. The host cell of claim 155, wherein the eukaryotic cell is a mammalian cell.

157. The host cell of claim 156, wherein the mammalian cell is a CHO cell.

158. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 106 to 144, a construct according to any one of claims 146 to 148, a polynucleotide according to claim 149, a vector according to any one of claims 150 to 153, or a host cell according to any one of claims 154 to 157, and a pharmaceutically acceptable carrier or excipient.

159. A method for modulating an immune response in a human subject, the method comprising administering to the subject an antibody or antigen-binding fragment thereof according to any one of claims 106 to 144, a construct according to any one of claims 146 to 148, a polynucleotide according to claim 149, a vector according to any one of claims 150 to 153, a host cell according to any one of claims 154 to 157, or a pharmaceutical composition according to claim 158.

160. The method of claim 159, wherein the method: (a) For the treatment of said human subjects with cell proliferation disorders, autoimmune diseases, infectious diseases, inflammatory diseases, neurological diseases, allergies, transplant rejection, or graft-versus-host disease; or (b) For transplantation or for regeneration of the tissues or organs of the human subject.

161. The method of claim 160, wherein the method is used to treat a cell proliferation disorder, and wherein the antibody or its antigen-binding fragment is: (a) A stimulatory antibody or its antigen-binding fragment that specifically binds to TNFRSF member proteins selected from the group consisting of: TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TNFR1, Fas, CD40, CD27, 4-1BB, OX40, GITR, and XEDAR; or (b) Antagonistic antibodies or their antigen-binding fragments that specifically bind to TNFRSF member proteins selected from the group consisting of: TRAMP, NGFR, TRAIL-R4, TNFR2, HVEM, CD30, TROY, and RELT.

162. The method according to any one of claims 159 to 161, wherein: (a) The serum or plasma level of the soluble TNFRSF member protein in the human subject is lower than or equal to a reference level of the soluble TNFRSF member protein, and the method comprises administering to the subject an agonistic antibody or an antigen-binding fragment thereof that specifically binds to the TNFRSF member protein; or (b) The serum or plasma level of the soluble TNFRSF member protein in the human subject is higher than the reference level of the soluble TNFRSF member protein, and the method comprises administering to the subject an antagonistic antibody or an antigen-binding fragment thereof that specifically binds to the TNFRSF member protein.

163. The method according to any one of claims 159 to 162, wherein the antibody or its antigen-binding fragment is administered to the human subject once weekly at a dose of about 0.8 mg / kg to about 1.2 mg / kg.

164. The method according to any one of claims 159 to 162, wherein the antibody or its antigen-binding fragment is administered to the human subject every two weeks at a dose of about 1.65 mg / kg to about 2.5 mg / kg.

165. The method according to any one of claims 159 to 162, wherein the antibody or its antigen-binding fragment is administered to the human subject at a dose of about 2.2 mg / kg to about 3.0 mg / kg once every three weeks.

166. The method according to any one of claims 159 to 162, wherein the antibody or its antigen-binding fragment is administered to the human subject once every four weeks at a dose of about 2.5 mg / kg to about 4.2 mg / kg.

167. A kit comprising an agent selected from the group consisting of: an antibody or antigen-binding fragment thereof according to any one of claims 106 to 144, a construct according to any one of claims 146 to 148, a polynucleotide according to claim 149, a vector according to any one of claims 150 to 153, a host cell according to any one of claims 154 to 157, or a pharmaceutical composition according to claim 158.

168. The kit of claim 167, wherein the kit further comprises: (a) Instructions for transfecting the vector into host cells; (b) Instructions for expressing the antibody or its antigen-binding fragment or the construct in host cells; (c) A reagent capable of expressing the antibody or its antigen-binding fragment or the construct in host cells; (d) Instructions for use in administering the drug to human subjects; (e) Instructions for use in the preparation or use of the pharmaceutical agent; and / or (f) Instructions for determining the level of the soluble TNFRSF member protein.

169. A method comprising: （a） (i) Detecting the level of soluble TNFRSF member proteins in a biofluid from a subject, wherein the level of the soluble TNFRSF member proteins in the biofluid is higher than a reference level for the soluble TNFRSF member proteins; and (ii) Administering to the subject at a predetermined dosing frequency one or more doses of an antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein, wherein the method is used to reduce the level of the soluble TNFRSF member protein; or （b） (i) Detecting the level of soluble TNFRSF member proteins in a biofluid from a subject, wherein the level of the soluble TNFRSF member proteins in the biofluid is normal or lower relative to a reference level; and (ii) Administering to the subject one or more doses of an agonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein at a given dosing frequency, wherein the method is used to increase the level of the soluble TNFRSF member protein.

170. The method of claim 169, wherein the reduced level of the soluble TNFRSF member protein in the human subject indicates the presence of insufficient T cell suppression and a need for more TNFR2 function, such as in the human subject’s cell proliferation disorder, autoimmune disease, inflammatory disease, neurological disease, allergy, transplant rejection, or graft-versus-host disease, optionally wherein the neurological disease is Alzheimer’s disease.

171. The method according to claim 169 or 170, wherein: (i) The method includes detecting the level of elevation of the soluble TNFRSF member protein in the subject's biofluid relative to a reference level, wherein the method further includes: -Increase the dose and / or frequency of administration of the antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein; or - Reduce the dose and / or frequency of administration of the agonist antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein; or (ii) The method comprises detecting a normal or reduced level of the soluble TNFRSF member protein in the subject's biological fluid relative to a reference level of the soluble TNFRSF member protein, wherein the method further comprises: -Increase the dose and / or frequency of administration of the agonist antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein; or - Reduce the dose and / or frequency of administration of the antagonistic antibody or its antigen-binding fragment that specifically binds to the TNFRSF member protein.

172. The method according to any one of claims 169 to 171, wherein the TNFRSF member protein is selected from the group consisting of: TNFR2, 4-1BB, CD27, CD30, CD40, DR6, EDAR, Fas, GITR, HVEM, LT-β receptor, NGFR, OPG, OX40, RANK, RELT (TNFRSF19L), TNFR1, TRAIL-R1 (TNFRSF10A), TRAIL-R2 (TNFRSF10B), TRAIL-R4, TRAMP, TROY, XEDAR, and DCR3, optionally wherein the TNFRSF member protein is TNFR2.

173. The method according to any one of claims 169 to 172, wherein the antagonistic antibody or the agonistic antibody or their antigen-binding fragments that specifically bind to the TNFRSF member protein are administered to the human subject at a dosing frequency of once weekly, once every two weeks, once every three weeks, or once every four weeks.

174. The method according to any one of claims 169 to 173, wherein the antagonistic antibody or the agonistic antibody or their antigen-binding fragments that specifically bind to the TNFRSF member protein are administered to the human subject at a dose of about 0.8 mg / kg to about 1.2 mg / kg, about 1.65 mg / kg to about 2.5 mg / kg, about 2.2 mg / kg to about 3.0 mg / kg, or about 2.5 mg / kg to about 4.2 mg / kg.

175. The method according to any one of claims 169 to 174, wherein the antagonistic antibody or the agonistic antibody or their antigen-binding fragments that specifically bind to the TNFRSF member protein are administered to the human subject in the following manner: (a) Once a week at a dose of about 0.8 mg / kg to about 1.2 mg / kg; (b) Once every two weeks at a dose of about 1.65 mg / kg to about 2.5 mg / kg; (c) Once every three weeks at a dose of about 2.2 mg / kg to about 3.0 mg / kg; or (d) Administer at a dose of approximately 2.5 mg / kg to approximately 4.2 mg / kg once every four weeks.

176. The method according to any one of claims 169 to 175, wherein the method further comprises reducing the dose range of the antagonistic antibody or the agonistic antibody or their antigen-binding fragments that specifically bind to the TNFRSF member protein.

177. The method according to any one of claims 169 to 176, wherein the method: (a) For the treatment of said human subjects with cell proliferation disorders, autoimmune diseases, infectious diseases, inflammatory diseases, neurological diseases, allergies, transplant rejection, or graft-versus-host disease; or (b) For transplantation or for regeneration of the tissues or organs of the human subject.

178. The method according to any one of claims 169 to 177, wherein the biological fluid is serum or plasma.

179. The method according to any one of claims 169 to 178, wherein the reference level of the soluble TNFRSF member protein is the level of the soluble TNFRSF in the biofluid of a healthy person.

180. The method according to any one of claims 169 to 179, wherein the one or more doses of the antibody or its antigen-binding fragment are administered to the human subject during one or more treatment periods, wherein: (a) Each dose contains: (i) about 0.1 mg to about 5000 mg of the antibody or its antigen-binding fragment; or (ii) the antibody or its antigen-binding fragment at a concentration of about 0.001 mg / kg to about 50 mg / kg; and / or (b) Each treatment period lasts for one, two, three, four, five, six, seven, eight or longer weeks.

181. The method according to any one of claims 169 to 180, wherein the administration frequency is once or more per month, every three weeks, every two weeks, weekly, every six days, every five days, every four days, every three days, every two days, or daily.

182. An antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, wherein the antibody or antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3, wherein: (a) The CDR-H1 contains the amino acid sequence GXTFXXY (SEQ ID NO: 1449), wherein each X is independently a naturally occurring amino acid; (b) The CDR-H2 comprises the amino acid sequences of NPNYDS (SEQ ID NO: 1414), NYDSTS (SEQ ID NO: 1420), or variants thereof having at most two conserved amino acid substitutions; (c) The CDR-H3 comprises the amino acid sequence of GNSWYFDV (SEQ ID NO: 1415) or a variant thereof having at most two conserved amino acid substitutions; (d) The CDR-L1 comprises the amino acid sequence of SASSSVRYMY (SEQ ID NO: 1416) or a variant thereof having at most two conserved amino acid substitutions; (e) The CDR-L2 comprises the amino acid sequence of LTSNLAS (SEQ ID NO: 1417) or a variant thereof having at most two conserved amino acid substitutions; and (f) The CDR-L3 comprises the amino acid sequence of QQWSSNPLT (SEQ ID NO: 1418) or a variant thereof having at most two conserved amino acid substitutions.

183. The antibody or antigen-binding fragment thereof according to claim 182, wherein the CDR-H1 comprises an amino acid sequence selected from the group consisting of: GYTFXXY (SEQ ID NO: 1450), GSTFXXY (SEQ ID NO: 1451), GTTFXXY (SEQ ID NO: 1452), GCTFXXY (SEQ ID NO: 1453), GNTFXXY (SEQ ID NO: 1454), GQTFXXY (SEQ ID NO: 1455), GXTFSXY (SEQ ID NO: 1456), GXTFCXY (SEQ ID NO: 1457), GXTFYXY (SEQ ID NO: 1458), GXTFNXY (SEQ ID NO: 1459), GXTFQXY (SEQ ID NO: 1460), and GXTFXDY (SEQ ID NO: 1461), wherein each X is independently a naturally occurring amino acid.

184. The antibody or antigen-binding fragment thereof according to claim 182 or 183, wherein the CDR-H1 comprises the amino acid sequence of GYTFTDY (SEQ ID NO: 1413) or a variant thereof having at most two conserved amino acid substitutions.

185. The antibody or antigen-binding fragment thereof according to any one of claims 182 to 184, wherein: (a) The CDR-H1 contains the amino acid sequence of GYTFTDY (SEQ ID NO: 1413); (b) The CDR-H2 contains the amino acid sequence of NPNYDS (SEQ ID NO: 1414); (c) The CDR-H3 contains the amino acid sequence of GNSWYFDV (SEQ ID NO: 1415); (d) The CDR-L1 contains the amino acid sequence SASSVRYMY (SEQ ID NO: 1416); (e) The CDR-L2 contains the amino acid sequence of LTSNLAS (SEQ ID NO: 1417); and (f) The CDR-L3 contains the amino acid sequence QQWSSNPLT (SEQ ID NO: 1418).

186. The antibody or antigen-binding fragment thereof according to any one of claims 182 to 184, wherein: (a) The CDR-H1 contains the amino acid sequence of GYTFTDY (SEQ ID NO: 1413); (b) The CDR-H2 contains the amino acid sequence of NYDSTS (SEQ ID NO: 1420); (c) The CDR-H3 contains the amino acid sequence of GNSWYFDV (SEQ ID NO: 1415); (d) The CDR-L1 contains the amino acid sequence SASSVRYMY (SEQ ID NO: 1416); (e) The CDR-L2 contains the amino acid sequence of LTSNLAS (SEQ ID NO: 1417); and (f) The CDR-L3 contains the amino acid sequence QQWSSNPLT (SEQ ID NO: 1418).

187. An antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, wherein the antibody or antigen-binding fragment comprises CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3, wherein: (a) The CDR-H1 comprises the amino acid sequence of GYTFTDYNLD (SEQ ID NO: 1425), DYIMH (SEQ ID NO: 1426) or variants thereof having at most two conserved amino acid substitutions; (b) The CDR-H2 comprises the amino acid sequence of DINPNYDS (SEQ ID NO: 1427), WVDPEYGSTDYAEKFKK (SEQ ID NO: 1428), or variants thereof having at most two conserved amino acid substitutions; (c) The CDR-H3 comprises the amino acid sequence of TSYSQKFRG (SEQ ID NO: 1429), DDGSYSPFEDY (SEQ ID NO: 1430) or variants thereof having at most two conserved amino acid substitutions; (d) The CDR-L1 comprises an amino acid sequence selected from the group consisting of: SASSSVRYMYWY (SEQ ID NO: 1431), KASENVVTYVS (SEQ ID NO: 1432), RSSQSLVHSNGNTYLH (SEQ ID NO: 1433), KASENVVTYVS (SEQ ID NO: 1434), QASQNINKYIA (SEQ ID NO: 1435), QNINKY (SEQ ID NO: 1436), and variants thereof having at most two conserved amino acid substitutions; (e) The CDR-L2 comprises an amino acid sequence selected from the group consisting of: LTSNLAS (SEQ ID NO: 1437), VTSNLAS (SEQ ID NO: 1438), LTSNLGS (SEQ ID NO: 1439), GASNRTY (SEQ ID NO: 1440), IKVSNRFS (SEQ ID NO: 1441), DPEYGS (SEQ ID NO: 1442), YTSTLES (SEQ ID NO: 1443), YTS, and variants thereof having at most two conserved amino acid substitutions; and (f) The CDR-L3 comprises an amino acid sequence selected from the group consisting of: QQRSNWP (SEQ ID NO: 1445), GQGYSYPYT (SEQ ID NO: 1446), SQTTHVPPT (SEQ ID NO: 1447), LQYVNLIT (SEQ ID NO: 1448), and variants thereof having at most two conserved amino acid substitutions.

188. The antibody or antigen-binding fragment thereof according to claim 187, wherein: (a) The CDR-H1 contains the amino acid sequence of GYTFTDYNLD (SEQ ID NO: 1425) or DYIMH (SEQ ID NO: 1426); (b) The CDR-H2 contains the amino acid sequence DINPNYDS (SEQ ID NO: 1427) or WVDPEYGSTDYAEKFKK (SEQ ID NO: 1428); (c) The CDR-H3 contains the amino acid sequence of TSYSQKFRG (SEQ ID NO: 1429) or DDGSYSPFEDY (SEQ ID NO: 1430); (d) The CDR-L1 comprises an amino acid sequence selected from the group consisting of: SASSSVRYMYWY (SEQ ID NO: 1431), KASENVVTYVS (SEQ ID NO: 1432), RSSQSLVHSNGNTYLH (SEQ ID NO: 1433), KASENVVTYVS (SEQ ID NO: 1434), QASQNINKYIA (SEQ ID NO: 1435), and QNINKY (SEQ ID NO: 1436); (e) The CDR-L2 comprises an amino acid sequence selected from the group consisting of: LTSNLAS (SEQ ID NO: 1437), VTSNLAS (SEQ ID NO: 1438), LTSNLGS (SEQ ID NO: 1439), GASNRTY (SEQ ID NO: 1440), IKVSNRFS (SEQ ID NO: 1441), DPEYGS (SEQ ID NO: 1442), YTSTLES (SEQ ID NO: 1443), and YTS; and (f) The CDR-L3 comprises an amino acid sequence selected from the group consisting of: QQRSNWP (SEQ ID NO: 1445), GQGYSYPYT (SEQ ID NO: 1446), SQTTHVPPT (SEQ ID NO: 1447) and LQYVNLIT (SEQ ID NO: 1448).

189. An antibody or an antigen-binding fragment thereof that specifically binds to human TNFR2, wherein the antibody or antigen-binding fragment comprises: (a) VH, wherein the VH has an amino acid sequence that is at least 80% identical to any of SEQ ID NO: 1486-1490; and (b) VL, wherein the VL has at least 80% identical amino acid sequence to any of SEQ ID NO: 1492-1497.

190. The antibody or antigen-binding fragment thereof according to claim 189, wherein the antibody or antigen-binding fragment thereof comprises: (a) VH, said VH having the amino acid sequence of any one of SEQ ID NO: 1486-1490; and (b) VL, wherein the VL has the amino acid sequence of any one of SEQ ID NO: 1492-1497.

191. The antibody or antigen-binding fragment thereof according to claim 189, wherein the antibody or antigen-binding fragment thereof comprises: (a) VH and VL, wherein VH has at least 80% of the same amino acid sequence as SEQ ID NO: 1488, and VL has at least 80% of the same amino acid sequence as SEQ ID NO: 1495; (b) VH and VL, wherein VH has at least 80% of the same amino acid sequence as SEQ ID NO: 1488, and VL has at least 80% of the same amino acid sequence as SEQ ID NO: 1497; (c) VH and VL, wherein VH has at least 80% of the same amino acid sequence as SEQ ID NO: 1489, and VL has at least 80% of the same amino acid sequence as SEQ ID NO: 1496; (d) VH and VL, wherein VH has at least 80% of the same amino acid sequence as SEQ ID NO: 1490, and VL has at least 80% of the same amino acid sequence as SEQ ID NO: 1496; or (e) VH and VL, wherein VH has at least 80% of the same amino acid sequence as SEQ ID NO: 1490, and VL has at least 80% of the same amino acid sequence as SEQ ID NO: 1497.

192. The antibody or antigen-binding fragment thereof according to claim 191, wherein the antibody or antigen-binding fragment thereof comprises: (a) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1488 and VL has the amino acid sequence of SEQ ID NO: 1495; (b) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1488 and VL has the amino acid sequence of SEQ ID NO: 1497; (c) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1489 and VL has the amino acid sequence of SEQ ID NO: 1496; (d) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1490 and VL has the amino acid sequence of SEQ ID NO: 1496; or (e) VH and VL, wherein VH has the amino acid sequence of SEQ ID NO: 1490 and VL has the amino acid sequence of SEQ ID NO: 1497.

193. The antibody or antigen-binding fragment thereof according to claim 189, wherein the antibody or antigen-binding fragment thereof comprises VH and VL, wherein the VH has at least 80% identical amino acid sequence to SEQ ID NO: 1488, and the VL has at least 80% identical amino acid sequence to SEQ ID NO: 1495.

194. The antibody or antigen-binding fragment thereof according to claim 193, wherein the antibody or antigen-binding fragment thereof comprises VH and VL, wherein the VH has the amino acid sequence of SEQ ID NO: 1488 and the VL has the amino acid sequence of SEQ ID NO: 1495.

195. The antibody or antigen-binding fragment thereof according to any one of claims 189 to 194, wherein the antibody or antigen-binding fragment thereof is a human antibody or antigen-binding fragment thereof.

196. The antibody or antigen-binding fragment thereof according to any one of claims 189 to 194, wherein the antibody or antigen-binding fragment thereof is an IgG antibody or antigen-binding fragment thereof.

197. The antibody or antigen-binding fragment thereof according to any one of claims 189 to 196, wherein the antibody or antigen-binding fragment thereof is a human IgG1 antibody, a human IgG2 antibody, a human IgG3 antibody or a human IgG4 antibody or an antigen-binding fragment thereof.

198. The antibody or antigen-binding fragment thereof according to claim 197, wherein the antibody or antigen-binding fragment thereof is a human IgG4 antibody or antigen-binding fragment thereof.

199. The antibody or antigen-binding fragment thereof according to any one of claims 189 to 198, wherein the antibody or antigen-binding fragment thereof further comprises a human IgG4 Fc domain.

200. The antibody or antigen-binding fragment thereof according to claim 199, wherein the human IgG4 Fc domain comprises a human IgG4 hinge region having an amino acid substitution S228P according to EU index number.

201. An antibody or antigen-binding fragment thereof that specifically binds to human TNFR2, wherein the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain having at least 80% identical amino acid sequence to any one of SEQ ID NO: 1498-1503; and (b) A light chain having at least 80% identical amino acid sequence to any of SEQ ID NO: 1504-1510.

202. The antibody or antigen-binding fragment thereof according to claim 201, wherein the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain having the amino acid sequence of any one of SEQ ID NO: 1498-1503; and (b) A light chain having an amino acid sequence of any one of SEQ ID NO: 1504-1510.

203. The antibody or antigen-binding fragment thereof according to claim 201, wherein the antibody or antigen-binding fragment thereof comprises: (a) A heavy chain and a light chain, wherein the heavy chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1498, and the light chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1504; (b) A heavy chain and a light chain, wherein the heavy chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1501, and the light chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1508; (c) A heavy chain and a light chain, wherein the heavy chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1501, and the light chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1510; (d) A heavy chain and a light chain, wherein the heavy chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1502, and the light chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1509; (e) A heavy chain and a light chain, wherein the heavy chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1503, and the light chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1509; or (f) A heavy chain and a light chain, wherein the heavy chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1503, and the light chain has at least 80% of the same amino acid sequence as SEQ ID NO: 1510.

204. The antibody or antigen-binding fragment thereof according to claim 203, wherein the antibody or antigen-binding fragment thereof comprises: (a) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1498 and the light chain has the amino acid sequence of SEQ ID NO: 1504; (b) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1501 and the light chain has the amino acid sequence of SEQ ID NO: 1508; (c) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1501 and the light chain has the amino acid sequence of SEQ ID NO: 1510; (d) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1502 and the light chain has the amino acid sequence of SEQ ID NO: 1509; (e) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1503 and the light chain has the amino acid sequence of SEQ ID NO: 1509; or (f) A heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 1503 and the light chain has the amino acid sequence of SEQ ID NO: 1510.

205. The antibody or antigen-binding fragment thereof according to claim 201, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, the heavy chain having at least 80% of the same amino acid sequence as SEQ ID NO: 1501, and the light chain having at least 80% of the same amino acid sequence as SEQ ID NO: 1508.

206. The antibody or antigen-binding fragment thereof according to claim 205, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, the heavy chain having the amino acid sequence of SEQ ID NO: 1501, and the light chain having the amino acid sequence of SEQ ID NO: 1508.

207. The antibody or antigen-binding fragment thereof according to any one of claims 201 to 206, wherein the antibody or antigen-binding fragment thereof is a human IgG4 antibody or antigen-binding fragment thereof.

208. The antibody or antigen-binding fragment thereof according to any one of claims 182 to 207, wherein the antibody or antigen-binding fragment thereof is an agonistic antibody or antigen-binding fragment thereof that specifically binds to human TNFR2.

209. The antibody or antigen-binding fragment thereof according to any one of claims 182 to 208, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of: monoclonal antibodies or antigen-binding fragments thereof, polyclonal antibodies or antigen-binding fragments thereof, human antibodies or antigen-binding fragments thereof, humanized antibodies or antigen-binding fragments thereof, primate-derived antibodies or antigen-binding fragments thereof, bispecific antibodies or antigen-binding fragments thereof, multispecific antibodies or antigen-binding fragments thereof, bivariate immunoglobulin domains, monovalent antibodies or antigen-binding fragments thereof, chimeric antibodies or antigen-binding fragments thereof, single-chain Fv molecules (scFv), bifunctional antibodies, trifunctional antibodies, antibody-like protein scaffolds, Fv fragments, Fab fragments, F(ab')2 molecules, and tandem scFv (taFv).

210. The antibody or antigen-binding fragment thereof according to claim 209, wherein the antibody or antigen-binding fragment thereof is a human antibody, a humanized antibody or a chimeric antibody or an antigen-binding fragment thereof.

211. A method for producing an antibody or an antigen-binding fragment thereof according to any one of claims 182 to 210, the method comprising expressing a polynucleotide encoding the antibody or an antigen-binding fragment thereof in a host cell and recovering the antibody or an antigen-binding fragment thereof from a host cell culture medium.

212. A construct comprising a first polypeptide domain and a second polypeptide domain, wherein the first polypeptide domain and the second polypeptide domain are each independently an antigen-binding fragment according to any one of claims 182 to 210.

213. The construct of claim 212, wherein the first polypeptide domain and the second polypeptide domain are linked by a covalent linker.

214. The construct of claim 213, wherein the covalent joint comprises an amide bond or a disulfide bond.

215. A polynucleotide encoding an antibody or antigen-binding fragment thereof according to any one of claims 182 to 210, or a construct according to any one of claims 212 to 214.

216. A vector comprising the polynucleotide according to claim 215.

217. The vector according to claim 216, wherein the vector is an expression vector or a viral vector.

218. The carrier according to claim 217, wherein: (a) The expression vector is a eukaryotic expression vector; or (b) The viral vector is selected from the group consisting of: adenovirus, retrovirus, poxvirus, adeno-associated virus, baculovirus, herpes simplex virus and vaccinia virus.

219. The carrier according to claim 218, wherein: (a) The adenovirus is serotype 1-60 adenovirus; (b) The adenovirus is serotype 5, 26, 35 or 48 adenovirus; (c) The retrovirus is a gamma-retrovirus or a lentivirus; or (d) The vaccinia virus is a modified Ankara vaccinia virus (MVA).

220. An isolated host cell comprising a vector according to any one of claims 216 to 219.

221. The host cell of claim 220, wherein the host cell is a prokaryotic cell or a eukaryotic cell.

222. The host cell of claim 221, wherein the eukaryotic cell is a mammalian cell.

223. The host cell of claim 222, wherein the mammalian cell is a CHO cell.

224. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 182 to 210, a construct according to any one of claims 212 to 214, a polynucleotide according to claim 215, a vector according to any one of claims 216 to 219, or a host cell according to any one of claims 220 to 223, and a pharmaceutically acceptable carrier or excipient.

225. A method for modulating an immune response in a human subject, the method comprising administering to the subject an antibody or antigen-binding fragment thereof according to any one of claims 182 to 210, a construct according to any one of claims 212 to 214, a polynucleotide according to claim 215, a vector according to any one of claims 216 to 219, a host cell according to any one of claims 220 to 223, or a pharmaceutical composition according to claim 224.

226. The method of claim 225, wherein the method: (a) For the treatment of said human subjects with cell proliferation disorders, autoimmune diseases, infectious diseases, inflammatory diseases, neurological diseases, allergies, transplant rejection, or graft-versus-host disease; or (b) For transplantation or for regeneration of the tissues or organs of the human subject.

227. The method of claim 226, wherein the method is used to treat cancer.

228. The method of any one of claims 225 to 227, wherein the serum or plasma level of soluble TNFR2 in the human subject is normal or lower than a reference level of soluble TNFR2, optionally wherein the method is used to increase the serum or plasma level of soluble TNFR2 in the human subject.

229. The method according to any one of claims 225 to 228, wherein the method further comprises: (a) Obtaining biofluids from the human subjects; (b) Detect the normal or reduced level of soluble TNFR2 in the subjects relative to a reference level of soluble TNFR2; and (c) Administer one or more doses of the antibody or its antigen-binding fragment to the subject at a certain dosing frequency.

230. The method of claim 229, wherein the method further comprises: (a) Detecting the normal or reduced level of soluble TNFR2 in the subject relative to a reference level of soluble TNFR2, and increasing the level of soluble TNFR2 by administering the antibody or its antigen-binding fragment at an increased dose and / or frequency; or (b) Detect the level of increase in soluble TNFR2 level in the subject relative to a soluble TNFR2 reference level, and increase the dose of the antibody or its antigen-binding fragment and / or the frequency of administration.

231. The method of claim 229 or 230, wherein the antibody or its antigen-binding fragment is administered to the human subject at a dosing frequency of once weekly, once every two weeks, once every three weeks, or once every four weeks.

232. The method according to any one of claims 229 to 231, wherein the antibody or its antigen-binding fragment is administered to the human subject at a dose of about 0.8 mg / kg to about 1.2 mg / kg, about 1.65 mg / kg to about 2.5 mg / kg, about 2.2 mg / kg to about 3.0 mg / kg, or about 2.5 mg / kg to about 4.2 mg / kg.

233. The method according to any one of claims 229 to 232, wherein the antibody or its antigen-binding fragment is administered to the human subject in the following manner: (a) Once a week at a dose of about 0.8 mg / kg to about 1.2 mg / kg; (b) Once every two weeks at a dose of about 1.65 mg / kg to about 2.5 mg / kg; (c) Once every three weeks at a dose of about 2.2 mg / kg to about 3.0 mg / kg; or (d) Administer at a dose of approximately 2.5 mg / kg to approximately 4.2 mg / kg once every four weeks.

234. The method according to any one of claims 229 to 233, wherein the method further comprises reducing the dose range of the antibody or its antigen-binding fragment.

235. A modified antibody or antigen-binding fragment thereof that specifically binds to tumor necrosis factor receptor 2 (TNFR2), wherein the modified antibody or antigen-binding fragment comprises a heavy chain variable region, a light chain variable region, and a modified human IgG4 constant region comprising a modified human IgG4 Fc hinge region, wherein the modified human IgG4 constant region comprises an amino acid sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 1547, wherein the modified human IgG4 Fc hinge region comprises an amino acid substitution S228P, and wherein the amino acid positions are according to EU index numbers.

236. The modified antibody or antigen-binding fragment thereof according to claim 235, wherein the TNFR2 is human TNFR2.

237. The modified antibody or antigen-binding fragment thereof according to claim 235, wherein the modified antibody or antigen-binding fragment thereof is a modified human IgG4 antibody or antigen-binding fragment thereof comprising a modified human IgG4 Fc domain, wherein the modified human IgG4 Fc domain comprises an amino acid substitution S228P.

238. The modified antibody or antigen-binding fragment thereof according to claim 237, wherein the modified human IgG4 constant region comprises the amino acid sequence of SEQ ID NO: 1547.

239. The modified antibody or antigen-binding fragment thereof according to claim 237 or 238, wherein the heavy chain variable region comprises heavy chain complementarity-determining regions 1 (CDR-H1), 1 CDR-H2, and 1 CDR-H3, and the light chain variable region comprises light chain complementarity-determining regions 1 (CDR-L1), 1 CDR-L2, and 1 CDR-L3, wherein CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 respectively comprise the amino acid sequences of SEQ ID NO: 1413, 1414 or 1420, 1415, 1416, 1417, and 1418.

240. The modified antibody or antigen-binding fragment thereof according to claim 239, wherein the heavy chain variable region comprises an amino acid sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 679, 681, 1485, 1486, 1487, 1488, 1489 or 1490, and the light chain variable region comprises an amino acid sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 680, 682, 1491, 1492, 1493, 1494 or 1495.

241. The modified antibody or antigen-binding fragment thereof according to claim 239 or 240, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 679, 681, 1485, 1486, 1487, 1488, 1489 or 1490, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 680, 682, 1491, 1492, 1493, 1494 or 1495.

242. The modified antibody or antigen-binding fragment thereof according to claim 241, wherein: (a) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 679, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 680; (b) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 681, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 682; or (c) The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 1488, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 1495.

243. The modified antibody or antigen-binding fragment thereof according to any one of claims 237 to 242, wherein the modified antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence having at least 95% identity with the sequence of SEQ ID NO: 1498, 1499, 1500, 1501, 1502 or 1503, and the light chain comprising an amino acid sequence having at least 95% identity with the sequence of SEQ ID NO: 1504, 1505, 1506, 1507 or 1508.

244. The modified antibody or antigen-binding fragment thereof according to claim 243, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 1498, 1499, 1500, 1501, 1502 or 1503, and the light chain comprises the amino acid sequence of SEQ ID NO: 1504, 1505, 1506, 1507 or 1508.

245. The modified antibody or antigen-binding fragment thereof according to claim 239, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 1488, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 1495, and the modified human IgG4 constant region comprises the amino acid sequence of SEQ ID NO: 1547.

246. The modified antibody or antigen-binding fragment thereof according to claim 239, wherein the modified antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, the heavy chain comprising the amino acid sequence of SEQ ID NO: 1501, and the light chain comprising the amino acid sequence of SEQ ID NO: 1508.

247. The modified antibody or antigen-binding fragment thereof according to any one of claims 237 to 246, wherein the modified antibody or antigen-binding fragment thereof is a monoclonal antibody.

248. The modified antibody or antigen-binding fragment thereof according to any one of claims 237 to 247, wherein the modified antibody or antigen-binding fragment thereof is a humanized antibody.

249. The modified antibody or antigen-binding fragment thereof according to any one of claims 235 to 248, wherein the modified human IgG4 Fc domain further comprises one or more amino acid modifications selected from the group consisting of: (i) L235E; (ii) D265A; (iii) G237A; (iv) E318A; (v)E233P; (vi) G236R and L328R; and (vii) N297G or N297Q; The amino acid positions are based on the EU index number.

250. The modified antibody or antigen-binding fragment thereof according to any one of claims 235 to 248, wherein the modified human IgG4 Fc domain further comprises one or more amino acid modifications selected from the group consisting of: (i) L235E; and (ii) F234A and L235A; The amino acid positions are based on the EU index number.

251. The modified antibody or antigen-binding fragment thereof according to any one of claims 237 to 250, wherein the modified antibody or antigen-binding fragment thereof is an agonist antibody.

252. The modified antibody or antigen-binding fragment thereof according to any one of claims 237 to 251, wherein the modified antibody or antigen-binding fragment thereof is formulated for subcutaneous administration.

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