Anti-IL-18BP antibody

Abstract

Antibodies that bind to interleukin-18 binding protein (IL-18BP), and related compositions, are provided that can be used in any of a variety of therapeutic and diagnostic methods, including the treatment or diagnosis of cancer and other diseases.

Description

[Background technology]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 437,526, filed January 6, 2023, U.S. Provisional Patent Application No. 63 / 590,348, filed October 13, 2023, and U.S. Provisional Patent Application No. 63 / 596,580, filed November 6, 2023, the contents of which are incorporated herein by reference in their entireties.

[0002] Interleukin-18 (IL-18) is an immunostimulatory cytokine with antitumor activity. This cytokine plays a central role in linking inflammatory immune responses with tumor progression. Recombinant human IL-18 has been evaluated as a cancer immunotherapy agent, but this approach has not been successful, at least in part due to a human feedback loop in which IL-18 administration increases the production of IL-18BP, which neutralizes the administered IL-18 (see, e.g., Robertson et al., Clinical Cancer Res. 12:4265-4273, 2006).

[0003] IL-18BP is a high-affinity IL-18 decoy receptor and is frequently upregulated in tumors. Research suggests that IL-18BP is a secreted immune checkpoint and a barrier to IL-18 immune activity (see, for example, Zhou et al., Nature. 583(7817):609-614, 2020). IL-18BP is thought to inhibit the pro-inflammatory activity of IL-18 by sequestering IL-18 from its cell surface receptor. The affinity of IL-18 for IL-18BP is higher than that of IL-18 for the IL-18 receptor, and IL-18BP is frequently present in greater amounts than IL-18, resulting in strict regulation. Furthermore, IL-18BP has been shown to balance immune responses, particularly Th1 and Th2, and plays an important role in autoimmune diseases (see, e.g., Park et al., Biomedicines. 10(7):1750, 2022). Thus, there is a need for agents and methods that specifically regulate the activity of IL-18BP, and such agents and methods are provided herein. Summary of the Invention

[0004] The present disclosure relates to antibodies that bind to interleukin-18 binding protein (IL-18BP), and related compositions thereof, which may be used in any of a variety of therapeutic and diagnostic methods, including the treatment or diagnosis of cancer and other diseases.

[0005] Aspects of the present disclosure include antibodies that bind to interleukin-18 binding protein (IL-18BP), wherein at least one antibody competes with IL-18 for binding to IL-18BP. In another aspect, the present disclosure provides antibodies that bind to IL-18BP and interfere with the binding of IL-18BP to IL-18. In another aspect, the present disclosure provides antibodies that bind to IL-18BP and are antagonists of IL-18BP, wherein the antibodies antagonize the binding activity between IL-18BP and IL-18. In some embodiments, the present disclosure provides antibodies that bind to preformed IL-18-IL-18BP complexes. In some embodiments, the present disclosure provides antibodies that bind to free IL-18BP. In some embodiments, the antibodies bind to a conformational epitope of IL-18BP. In some embodiments, the antibody binds to two or more of amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In some embodiments, the antibody binds to amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In some embodiments, the antibody binds to a linear epitope of IL-18BP. In some embodiments, the antibody binds to the binding interface between IL-18 and mature IL-18BP. In some embodiments, the antibody binds to amino acid residues S75, H79, T116, and S119 of SEQ ID NO: 372.

[0006] In some embodiments, the antibody binds to human IL-18BP and cynomolgus IL-18BP, but not to mouse IL-18BP, hi some embodiments, the antibody binds to human IL-18BP, cynomolgus IL-18BP, and mouse IL-18BP.

[0007] In some embodiments, the present disclosure also includes antibodies that bind to interleukin-18 binding protein (IL-18BP), wherein at least one antibody has a V complementarity determining region selected from Table A1. H CDR1 sequence, V HCDR2 sequence, and V H A heavy chain variable region (V) containing CDR3 sequences and variants thereof that specifically bind to IL-18BP. H ), and a V of a complementarity-determining region selected from Table A1 L CDR1 sequence, V L CDR2 sequence, and V L A light chain variable region (V) containing CDR3 sequences and variants thereof that specifically bind to IL-18BP. L ) is included.

[0008] In some embodiments, V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V H has at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions. L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V L has at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

[0009] Also included are isolated polynucleotides encoding the anti-IL-18BP antibodies described herein, expression vectors containing the isolated polynucleotides, and isolated host cells containing the vectors. Also provided are one or more isolated polynucleotides encoding the anti-IL-18BP antibodies described herein. For example, the V of the antibodies disclosed herein H A first polynucleotide encoding a region and a V region of the antibody disclosed herein L A second polynucleotide encoding the region is provided herein.

[0010] Certain embodiments include pharmaceutical compositions comprising an anti-IL-18BP antibody described herein and a pharmaceutically acceptable carrier. In some embodiments, the composition is optionally a sterile injectable solution suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

[0011] Also included are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein. In some embodiments, the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, optionally a proliferative disease or disorder selected from lymphoproliferative disorders, myeloproliferative disorders, proliferative enterocolitis, proliferative diabetic retinopathy, and proliferative renal diseases. In some embodiments, the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18. In some embodiments, the cancer is selected from the group consisting of bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (liver cell carcinoma), and / or hepatocellular carcinoma (liver cell carcinoma). and / or ovarian cancer), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, urothelial cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer.

[0012] Also included is a method of screening anti-IL-18BP antibodies for the ability to block or inhibit binding between IL-18 and IL-18BP, the method comprising: (a) determining the binding affinity of the antibody for (i) IL-18BP alone and (ii) a low-IL-18 fusion protein, the low-IL-18 fusion protein comprising IL-18 fused to IL-18BP via a flexible linker (and optionally a protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein is linked to the IL-18BP portion of the fusion protein so as to sterically block the IL-18-binding site of the IL-18BP portion of the fusion protein; (b) comparing the binding affinity of (i) with the binding affinity of (ii); and (c) identifying or selecting the antibody as having the ability to block or inhibit binding between IL-18 and IL-18BP if the binding affinity of (i) is significantly stronger than the binding affinity of (ii).

[0013] Some aspects of the invention include isolated low IL-18 fusion proteins comprising, from N- to C-terminal, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP. In some embodiments, the low IL-18 fusion protein comprises, consists of, or consists essentially of an amino acid sequence that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1.

[0014] Also included are methods of stimulating an immune response in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein. In some embodiments, the immune response is an IL-18-mediated immune response. In certain embodiments, the IL-18-mediated immune response comprises induction of IFN-gamma, CXCL10, and / or TNFα in the subject. [Brief explanation of the drawings]

[0015] [Figure 1A-B] 1A-1B show the antibody family and IL-18-reduced design of the present disclosure. FIG. 1A shows the antibody family of the present disclosure. FIG. 1B shows a schematic of an exemplary IL-18-reduced expression cassette. Landmarks in the gene from the N' to C' terminus include the osteonectin signal peptide, the human IL-18 coding region, a flexible gly-ser linker interrupted by a tobacco etch virus (TEV) protease cleavage site, the human IL-18BP coding sequence, and a 6x HIS tag. [Figure 1C] Figure 1C shows the family of antibodies of the present disclosure and the design of low IL-18. Figure 1C shows an outline of a model derived from the crystal structure of human IL-18 complexed with ectromelia virus IL-18BP (shown). The N-terminus of IL-18BP and the C-terminus of IL-18 are indicated by boxes and arrows. [Figure 2A] Figure 2A shows the effect of anti-IL-18BP antibody on IL-18-mediated IFNγ induction in healthy human peripheral blood mononuclear cells (PBMCs) in vitro. Anti-IL-18BP antibody increased IFNγ in a dose-dependent manner. [Figure 2B] Figure 2B shows the effect of anti-IL-18BP antibody on IL-18-mediated IFNγ induction in healthy human peripheral blood mononuclear cells (PBMCs) in vitro. Anti-IL-18BP antibody increased IFNγ in a dose-dependent manner. [Figure 2C] Figure 2C shows the effect of anti-IL-18BP antibody on IL-18-mediated IFNγ induction in healthy human peripheral blood mononuclear cells (PBMCs) in vitro. Anti-IL-18BP antibody increased IFNγ in a dose-dependent manner. [Figure 2D] Figure 2D shows the effect of anti-IL-18BP antibody on IL-18-mediated IFNγ induction in healthy human peripheral blood mononuclear cells (PBMCs) in vitro. Anti-IL-18BP antibody increased IFNγ in a dose-dependent manner. [Figure 2E]Figure 2E shows the effect of anti-IL-18BP antibody on IL-18-mediated IFNγ induction in healthy human peripheral blood mononuclear cells (PBMCs) in vitro. Anti-IL-18BP antibody increased IFNγ in a dose-dependent manner. [Figure 2F] Figure 2F shows the effect of anti-IL-18BP antibody on IL-18-mediated IFNγ induction in healthy human peripheral blood mononuclear cells (PBMCs) in vitro. Anti-IL-18BP antibody increased IFNγ in a dose-dependent manner. [Figure 2G] Figure 2G shows the effect of anti-IL-18BP antibody on IL-18-mediated IFNγ induction in healthy human peripheral blood mononuclear cells (PBMCs) in vitro. Anti-IL-18BP antibody increased IFNγ in a dose-dependent manner. [Figure 3A] Figure 3A shows the ability of anti-IL-18BP antibodies to release IL-18 from pre-complexes with IL-18BP. After IL-18 was allowed to form a pre-complex with IL-18BP, anti-IL-18BP antibodies were added to human PBMCs. This resulted in a dose-dependent increase in IFNγ. [Figure 3B] Figure 3B shows the ability of anti-IL-18BP antibodies to release IL-18 from pre-complexes with IL-18BP. After IL-18 was allowed to form a pre-complex with IL-18BP, anti-IL-18BP antibodies were added to human PBMCs. This resulted in a dose-dependent increase in IFNγ. [Figure 3C] Figure 3C shows the ability of anti-IL-18BP antibodies to release IL-18 from pre-complexes with IL-18BP. After IL-18 was allowed to form a pre-complex with IL-18BP, anti-IL-18BP antibodies were added to human PBMCs. This resulted in a dose-dependent increase in IFNγ. [Figure 3D] Figure 3D shows the ability of anti-IL-18BP antibodies to release IL-18 from pre-complexes with IL-18BP. After IL-18 was allowed to form a pre-complex with IL-18BP, anti-IL-18BP antibodies were added to human PBMCs. This resulted in a dose-dependent increase in IFNγ. [Figure 3E]Figure 3E shows the ability of anti-IL-18BP antibodies to release IL-18 from pre-complexes with IL-18BP. After IL-18 was allowed to form a pre-complex with IL-18BP, anti-IL-18BP antibodies were added to human PBMCs. This resulted in a dose-dependent increase in IFNγ. [Figure 3F] Figure 3F shows the ability of anti-IL-18BP antibodies to release IL-18 from pre-complexes with IL-18BP. After IL-18 was allowed to form a pre-complex with IL-18BP, anti-IL-18BP antibodies were added to human PBMCs. This resulted in a dose-dependent increase in IFNγ. [Figure 3G] Figure 3G shows the ability of anti-IL-18BP antibodies to release IL-18 from pre-complexes with IL-18BP. After IL-18 was allowed to form a pre-complex with IL-18BP, anti-IL-18BP antibodies were added to human PBMCs. This resulted in a dose-dependent increase in IFNγ. [Figure 4] FIG. 4 shows the ability of anti-human IL-18BP antibodies to elicit IL-18-induced production of the cytokine CCL2 in human PBMCs by ELISA. [Figure 5A] Figure 5A shows the results of an assay of cynomolgus monkey PBMCs. Figure 5A shows the effect of anti-IL-18BP antibodies on IL-18-mediated IFNγ induction in vitro in cynomolgus monkey PBMCs. [Figure 5B] Figure 5B shows the results of an assay of cynomolgus monkey PBMCs, demonstrating the ability of anti-IL-18BP antibodies to release IL-18 from pre-existing complexes with IL-18BP via IFNγ release. [Figure 6] FIG. 6 shows the ability of IL-18BP antibodies to disrupt the IL-18:IL-18BP complex and induce IFNγ secretion from NK cells. DETAILED DESCRIPTION OF THE INVENTION

[0016] The present disclosure relates to antibodies that specifically bind to interleukin-18 binding protein (IL-18BP). Some embodiments include certain humanized antibodies that can bind to IL-18BP and block or reduce the inhibitory binding of IL-18BP to its ligand, IL-18, thereby increasing IL-18-mediated downstream signaling. Thus, in certain embodiments, anti-IL-18BP antibodies are antagonists or inhibitors of IL-18BP.

[0017] The IL-18BP antagonist antibodies described herein are useful for the treatment and prevention of various diseases and conditions, such as cancer. Accordingly, some embodiments relate to the use of anti-IL-18BP antibodies for the diagnosis, evaluation, and treatment of diseases and conditions, including those associated with IL-18 and / or associated with the activity or aberrant expression of IL-18BP.

[0018] The practice of the present disclosure will employ, unless specifically indicated to the contrary, conventional methods of virology, immunology, microbiology, molecular biology, and recombinant DNA technology within the skill of the art, many of which are described below for illustrative purposes. Such techniques are fully explained in the literature. See, for example, Current Protocols in Molecular Biology or Current Protocols in Immunology, John Wiley & Sons, New York, NY (2009); Ausubel et al., Short Protocols in Molecular Biology, 3 rded., Wiley & Sons, 1995; Sambrook and Russell, Molecular Cloning: A Laboratory Manual (3rd Edition, 2001); Maniatis et al. Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., 1984); Nucleic Acid Hybridization (B. Hames & S. Higgins, eds., 1985); Transcription and Translation (B. Hames & S. Higgins, eds., 1984); Animal Cell Culture (R. Freshney, ed., 1986); Perbal, A Practical Guide to Molecular Cloning (1984), and other similar references.

[0019] definition Unless otherwise defined, all technical terms, notations, and other technical and scientific terms or terminology used herein are intended to have the same meaning as commonly understood by those skilled in the art to which the claimed subject matter pertains. In some instances, terms having commonly understood meanings are defined herein for clarity and / or ready reference. The inclusion of such definitions herein should not necessarily be interpreted as representing a substantial departure from the definitions commonly understood in the art.

[0020] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the content clearly dictates otherwise.

[0021] As used herein, the term "about" is understood by those of ordinary skill in the art and varies to some extent depending on the context in which the term is used. In some embodiments, when referring to a measurable value, such as an amount, a period of time, etc., the term "about" is intended to encompass variations accepted in the art based on the standard error in making such measurements. In some embodiments, when referring to such values, the term "about" is intended to encompass a ±10% variation from the specified value.

[0022] As used herein, the term "antibody" includes not only intact polyclonal or monoclonal antibodies, but also antigen-binding fragments thereof (e.g., dAb, Fab, Fab', F(ab')2, Fv, etc.), single chain (scFv), synthetic variants thereof, naturally occurring variants, fusion proteins comprising an antibody portion with an antigen-binding fragment of the required specificity, humanized antibodies, chimeric antibodies, and any other modified configuration of an immunoglobulin molecule that contains an antigen-binding site or fragment (epitope-recognition site) of the required specificity. Specific features and characteristics of antibodies (and antigen-binding fragments thereof) are described in more detail herein.

[0023] As used herein, the term "antigen-binding fragment" refers to a polypeptide fragment that contains at least one CDR of an immunoglobulin heavy and / or light chain and that binds to an antigen of interest. In this regard, an antigen-binding fragment of an antibody described herein is a V CDR from an antibody that binds to a target molecule. H and V L In certain embodiments, the antigen-binding fragments of the present disclosure may comprise one, two, three, four, five, or all six CDRs of the V of an antibody disclosed herein. H Array and V L The sequence includes all six CDRs.

[0024] The binding properties of antibodies and their antigen-binding fragments can be quantified using methods well known in the art (see Davies et al., Annual Rev. Biochem. 59:439-473, 1990). In certain embodiments, the antibodies described herein comprise heavy and light chain complementarity-determining regions (CDRs), each of which is interposed between a set of heavy and light chain framework regions (FRs), which provide support for the CDRs and define the spatial relationship of the CDRs to each other. As used herein, the term "CDR set" refers to the three hypervariable regions of a heavy or light chain V region. Proceeding from the N-terminus of the heavy or light chain, these regions are designated "CDR1," "CDR2," and "CDR3," respectively. The antigen-binding site therefore comprises six CDRs, including a CDR set from each of the heavy and light chain V regions. A polypeptide comprising a single CDR (e.g., CDR1, CDR2, or CDR3) is referred to herein as a "molecular recognition unit." Crystallographic analysis of numerous antigen-antibody complexes has demonstrated that the amino acid residues of the CDRs form extensive contacts with the bound antigen, with the most extensive antigen contacts being with the heavy chain CDR3. Thus, the molecular recognition unit is primarily responsible for the specificity of the antigen-binding site.

[0025] The structure and location of immunoglobulin variable domains can be determined by reference to Kabat, EA et al., Sequences of Proteins of Immunological Interest. 4th Edition. U.S. Department of Health and Human Services, 1987, and updates thereof.

[0026] Also included are monoclonal antibodies, which refer to homogeneous antibody populations composed of amino acids (natural and non-natural) involved in selective binding of an epitope. The term "monoclonal antibody" encompasses not only intact and full-length monoclonal antibodies, but also fragments thereof (e.g., Fab, Fab', F(ab')2, Fv, etc.), single chain (scFv), variants thereof, fusion proteins containing the antigen-binding portion, humanized monoclonal antibodies, chimeric monoclonal antibodies, and any other modified configuration of an immunoglobulin molecule containing an antigen-binding fragment (epitope recognition site) of the required specificity and ability to bind to the epitope. It is not intended to be limited with regard to the source of the antibody or the manner in which it is made (e.g., by hybridoma, phage selection, recombinant expression, transgenic animals). The term includes whole immunoglobulins, as well as fragments such as those described above under the definition of "antibody."

[0027] In certain embodiments, antibodies are made human-like, for example, by generating chimeric antibodies. Chimeric antibodies are generally prepared using recombinant techniques and have an antigen-binding site derived from an immunoglobulin from a non-human species, with the remaining immunoglobulin structure of the molecule based on the structure and / or sequence of a human immunoglobulin. The antigen-binding site may comprise either a complete variable domain fused onto a constant domain, or only a CDR (whole or part) grafted onto suitable framework regions in the variable domain. The epitope-binding site may be wild-type or modified by one or more amino acid substitutions. This eliminates the constant region as an immunogen in human individuals, although the possibility of an immune response to the foreign variable region remains (LoBuglio et al., PNAS USA 86:4220-4224, 1989; Queen et al., PNAS USA. 86:10029-10033, 1988; Riechmann et al., Nature. 332:323-327, 1988). Exemplary methods for antibody humanization include those described in US Pat. No. 7,462,697.

[0028] Another approach focuses not only on providing human-derived constant regions, but also on modifying the variable regions and reshaping them as closely as possible to human form. Both heavy and light chain variable regions are known to contain three complementarity-determining regions (CDRs) that vary in response to the epitope in question and determine binding ability, flanked by four framework regions (FRs) that are relatively conserved in a given species and presumably provide a scaffold for the CDRs. When a non-human antibody is prepared for a specific epitope, the variable region can be reshaped or humanized by grafting CDRs derived from the non-human antibody onto the FRs present in the modified human antibody. Application of this approach to various antibodies has been reviewed by Sato et al., Cancer Res. 53:851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J. Immunol. 148:1149-1154, 1992. In some embodiments, a humanized antibody retains all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs from the mouse antibody). In some embodiments, only a portion of the CDR sequences are transplanted from a non-human antibody (Bowers et al., J. Biol. Chem. 288:7688-7696, 2013).In certain embodiments, a humanized antibody has one or more CDRs (one, two, three, four, five, six) that are altered relative to the original antibody, also referred to as one or more CDRs "derived from" one or more CDRs of the original antibody.

[0029] In certain embodiments, the antibody is a chimeric antibody. In this regard, a chimeric antibody is composed of an antigen-binding fragment of an antibody operably linked or otherwise fused to a heterologous Fc portion of a different antibody. In certain embodiments, the Fc domain or heterologous Fc domain is of human origin. In certain embodiments, the Fc domain or heterologous Fc domain is of murine origin. In other embodiments, the heterologous Fc domain can be from an Ig class different from that of the parent antibody, including IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), and IgM. In further embodiments, the heterologous Fc domain can be composed of CH2 and CH3 domains derived from one or more of the different Ig classes. As described above for humanized antibodies, an antigen-binding fragment of a chimeric antibody can include only one or more of the CDRs of an antibody described herein (e.g., one, two, three, four, five, or six CDRs of an antibody described herein), or can include the entire variable domain (VL, VH, or both).

[0030] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer, or group of elements or integers, but not the exclusion of any other element or integer or group of elements or integers.

[0031] "Consisting of" means including, but not limited to, everything that follows the phrase "consisting of." Thus, the phrase "consisting of" indicates that the listed elements are required or essential, and that no other elements will be present. By "consisting essentially of," it is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or function specified in this disclosure for the listed elements. Thus, the phrase "consisting essentially of" indicates that the listed elements are required or essential, but that other elements are optional and may or may not be present depending on whether they substantially affect the activity or function of the listed elements.

[0032] "Immune response" refers to any immunological response derived from the immune system, including responses from the cellular and humeral, innate, and adaptive immune systems. Exemplary cellular immune cells include, for example, lymphocytes, macrophages, T cells, B cells, NK cells, neutrophils, eosinophils, dendritic cells, mast cells, monocytes, and all subsets thereof. Cellular responses include, for example, effector functions, cytokine release, phagocytosis, efferocytosis, translocation, trafficking, proliferation, differentiation, activation, suppression, cell-cell interactions, apoptosis, and the like. Humoral responses include, for example, IgG, IgM, IgA, IgE, responses, and their corresponding effector functions.

[0033] "Expression control sequences" include regulatory sequences of nucleic acids or corresponding amino acids, such as promoters, leaders, enhancers, introns, recognition motifs for RNA or DNA binding proteins, polyadenylation signals, terminators, internal ribosome entry sites (IRES), secretion signals, subcellular localization signals, and the like, which can affect the transcription or translation of coding sequences in host cells, or the intracellular or cellular location. Exemplary expression control sequences are described in Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990).

[0034] The term "isolated" polypeptide or protein, as referred to herein, means that the subject protein (1) is free from at least some other proteins with which it would typically be found in nature; (2) is essentially free from other proteins from the same source, e.g., from the same species; (3) is expressed by cells from a different species; (4) is separated from at least about 50 percent of the polynucleotides, lipids, carbohydrates, or other materials with which it is naturally associated; (5) is not associated (by covalent or non-covalent interactions) with portions of proteins with which it is naturally associated; (6) is operably associated (by covalent or non-covalent interactions) with polypeptides with which it is not naturally associated; or (7) does not occur in nature. Such isolated proteins may be encoded by genomic DNA, cDNA, mRNA, or other RNA, may be of synthetic origin, or any combination thereof. In certain embodiments, an isolated protein is substantially free from proteins or polypeptides found in its natural environment or other contaminants that would interfere with its use (therapeutic, diagnostic, preventative, research, or otherwise).

[0035] Certain embodiments include biologically active "variants" and "fragments" of the polypeptides (e.g., antibodies) described herein, as well as polynucleotides encoding the same. "Variant" includes one or more substitutions, additions, deletions, and / or insertions relative to a reference polypeptide or polynucleotide (see, e.g., Tables and Sequence Listing). A variant polypeptide or polynucleotide comprises an amino acid or nucleotide sequence with at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity or similarity or homology to a reference sequence, as described herein, and substantially retains the activity of the reference sequence. Also included are sequences that consist of a reference sequence or that differ from a reference sequence by the addition, deletion, insertion, or substitution of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, or more amino acids or nucleotides, and that substantially retain the activity of the reference sequence. In certain embodiments, the additions or deletions include C-terminal and / or N-terminal additions and / or deletions.

[0036] The term "sequence identity" or, as used herein, includes, for example, "at least 50% identical sequence", refers to the degree to which sequences are identical on a nucleotide-by-nucleotide basis or on an amino acid-by-amino acid basis over a comparison window.Thus, "sequence identity percentage" can be calculated by: comparing two optimally aligned sequences over a comparison window; determining the number of positions where identical nucleic acid bases (e.g., A, T, C, G, I) or identical amino acid residues (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys and Met) occur in both sequences, resulting in the number of identical positions; dividing the number of identical positions by the total number of positions in the comparison window (i.e., window size), and multiplying the result by 100 to obtain the percentage of sequence identity. Optimal alignment of sequences for aligning a comparison window can be performed by computerized implementations of algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Drive, Madison, Wis., USA), or by inspection and best alignment (i.e., resulting in the highest percentage of homology over the comparison window) generated by any of a variety of selected methods. Reference can also be made to the BLAST family of programs, for example, as disclosed by Altschul et al., Nucl. Acids Res. 25:3389, 1997.

[0037] The terms "individual," "subject," and "person" are used interchangeably herein and refer to any subject for whom treatment or therapy is desired. The subject may be a mammalian subject. Mammalian subjects include, for example, humans, non-human primates, rodents (e.g., rats, mice), lagomorphs (e.g., rabbits), ungulates (e.g., cows, sheep, pigs, horses, goats, etc.), and the like. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human primate, e.g., a cynomolgus monkey. In some embodiments, the subject is a companion animal (e.g., a cat, a dog).

[0038] As used herein, the term "therapeutically effective amount," "therapeutic dose," "prophylactically effective amount," or "diagnostically effective amount" is the amount of an agent (e.g., an anti-IL-18BP antibody, an immunotherapeutic agent) required to elicit a desired biological response following administration.

[0039] As used herein, "treatment" of a subject (e.g., a mammal, such as a human primate or non-human primate) or cell is any type of intervention used in an attempt to alter the natural course of a disease or disorder. Treatment includes, but is not limited to, the administration of a pharmaceutical composition and can be performed either prophylactically or following the initiation of a pathological event or contact with a pathogenic agent. Also included is prophylactic treatment, which can be directed at reducing the rate of progression of the disease or condition being treated, delaying the onset of the disease or condition, or reducing the severity of its onset. "Treatment" or "prevention" does not necessarily indicate complete eradication, cure, or prevention of the disease or condition, or its associated symptoms.

[0040] Anti-IL-18BP antibody Certain embodiments include antibodies that bind to IL-18BP. In some embodiments, the antibodies modulate (e.g., interfere with, antagonize, inhibit) the binding of IL-18BP to its ligand, interleukin-18 (IL-18). In certain embodiments, the antibodies bind to the complementarity-determining region V H CDR1, VH CDR2, and V H The heavy chain variable region (V H ), and complementarity-determining region V L CDR1, V L CDR2, and V L The light chain variable region (V L ) characterized by or including them. H Array, V H CDR1 sequence, V H CDR2 sequence, V H CDR3 sequence, V L Array, V L CDR1 sequence, V L CDR2 sequence, and V L The CDR3 sequences are provided in Tables A1 and A2 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7]

[0041] Thus, in certain embodiments, the antibody comprises a V complementarity determining region selected from Table A1. H CDR1 sequence, V H CDR2 sequence, and V H V containing the CDR3 sequence H sequences, and variants thereof that bind to IL-18BP, and V of the complementarity determining region selected from Table A1L CDR1 sequence, V L CDR2 sequence, and V L V containing the CDR3 sequence L In certain embodiments, the antibody comprises the sequence V H CDR1 sequence, V H CDR2 sequence, and V H V containing the CDR3 sequence H Sequence, and V L CDR1 sequence, V L CDR2 sequence, and V L V containing the CDR3 sequence L sequences, in which all of the CDR sequences are derived from one named antibody (e.g., SA04a) in Table A1.

[0042] In certain embodiments, the CDR sequences are as follows: V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 4 to 6, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 7 to 9, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 10 to 12, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 13 to 15, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 16 to 18, respectively; V H CDR1 sequence, VH CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 19 to 21, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 22 to 24, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 25 to 27, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 28 to 30, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 31 to 33, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 34 to 36, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 37 to 39, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 40 to 42, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 43 to 45, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 46 to 48, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 49 to 51, respectively, and V LCDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 52 to 54, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 55 to 57, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 58 to 60, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 61 to 63, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 64 to 66, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 67 to 69, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 70 to 72, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 73 to 75, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 76 to 78, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 79 to 81, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 82 to 84, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 85 to 87, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 88 to 90, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 91 to 93, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 94 to 96, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 97 to 99, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 100 to 102, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 103 to 105, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 106-108, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 109 to 111, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; V H CDR1 sequence, V H CDR2 sequence, and VH The CDR3 sequences include SEQ ID NOs: 115 to 117, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 121 to 123, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 124 to 126, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 127 to 129, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 130 to 132, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 133 to 135, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 136 to 138, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 139 to 141, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 142 to 144, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 145 to 147, respectively, and V LCDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 148 to 150, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 151 to 153, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 154 to 156, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 157 to 159, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 160 to 162, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 163 to 165, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 169 to 171, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 172 to 174, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 175 to 177, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V Lthe CDR3 sequences comprise SEQ ID NOs: 178 to 180, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 181 to 183, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 184 to 186, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 187 to 189, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 190 to 192, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 193 to 195, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 199 to 201, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 205 to 207, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 208-210, respectively; V HCDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 211 to 213, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 214-216, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 217 to 219, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 223 to 225, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 229 to 231, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 232 to 234, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 235 to 237, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; V H CDR1 sequence, V H CDR2 sequence, and V HThe CDR3 sequences include SEQ ID NOs: 241 to 243, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 244 to 246, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 247 to 249, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 250 to 252, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 253 to 255, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 256 to 258, respectively; V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 259 to 261, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L The CDR3 sequences comprise SEQ ID NOs: 262 to 264, respectively.

[0043] Also included are variants of the aforementioned CDRs. Exemplary variants bind to IL-18BP and have one or more of the individual CDRs, e.g., a V or VL as described herein. H CDR1, V H CDR2, V H CDR3, V L CDR1, V L CDR2, and / or V L Any one or more of the CDR3 sequences may have a total of 1, 2, or 3 changes. Exemplary "changes" include amino acid substitutions, additions, and deletions.

[0044] Exemplary V H Array and V L Sequence, and V H / V L The sequence pairs are provided below in Table A2 (CDRs are underlined). [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6]

[0045] Thus, in certain embodiments, the antibody binds to IL-18BP and is selected from Table A2. H Array and corresponding V L In certain embodiments, V H includes a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, e.g., V H has at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in one or more framework regions. L includes a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, e.g., V L has at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in one or more framework regions. Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and V H In certain embodiments, the V region is derived from the same named antibody (e.g., SA04a antibody). H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and is derived from the same one named antibody (e.g., SA04a antibody) as the VH region, where no changes are present in the underlined CDRs of Table A2. Thus, the antibody comprises a VH region at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to each sequence from one named antibody (e.g., SA04a) of Table A2. H Array and V L Alternatively, the antibody may comprise a sequence, in which case the antibody comprises the CDRs of one of the named antibodies (e.g., SA04a) listed in Table A1.

[0046] In some embodiments, the V of the antibody H and V L is as follows: V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 265, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 266; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 267, and V Lcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 268; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 269, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 270; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 271, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 272; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 273, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 274; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 275, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 276; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and V Lcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 281, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 282; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 283, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 284; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 285, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 286; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 287, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 288; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 289, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 290; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 291, and V Lcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 292; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 293, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 294; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 295, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 296; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 297, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 298; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 299, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 300; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 301, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 302; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 303, and V Lcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 304; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 305, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 306; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 307, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 308; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 309, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 310; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 311, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 312; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 313, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 314; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 315, and V Lcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 316; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 317, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 318; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 319, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 320; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 321, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 322; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 323, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 324; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 325, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 326; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 327, and V Lcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 328; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 329, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 330; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 331, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 332; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 335, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 336; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 337, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 338; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 339, and V Lcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 340; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 341, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 342; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 343, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 344; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 345, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 346; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 347, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 348; V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 349, and V L comprises a sequence that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 350; or V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 351, and V Lcomprises a sequence that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:352.

[0047] Also, variants thereof that bind to IL-18BP, such as the aforementioned V H Sequence and / or V L These are variants having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in any one or more framework regions of the sequence. Exemplary "changes" include amino acid substitutions, additions, and deletions.

[0048] As mentioned above, the antibodies described herein bind to IL-18BP. In certain embodiments, the antibodies bind to human IL-18BP, cynomolgus monkey IL-18BP, and / or mouse IL-18BP, or a region, fragment, or epitope thereof.

[0049] Human interleukin-18 binding protein, or IL-18BP, is encoded by the IL18BP gene (Gene ID: 10068; and UniProt: O95998) and has at least three isoforms. In some embodiments, an antibody of the present disclosure binds to isoform A of IL-18BP. In some embodiments, an antibody of the present disclosure binds to isoform B of IL-18BP. In some embodiments, an antibody of the present disclosure binds to both isoform A and isoform C of IL-18BP. In some embodiments, an antibody of the present disclosure binds to all isoforms of IL-18BP. It is an inhibitor of the early Th1 cytokine response and the pro-inflammatory cytokine IL-18. For example, IL-18BP binds to IL-18 and inhibits the binding of IL-18 to its receptor, thereby inhibiting IL-18-induced IFN-gamma production, among other IL-18 signaling activities. The amino acid sequences of human, cynomolgus monkey, and mouse IL-18BP isoforms are presented below in Table B1. The signal peptide is underlined in the table below. [Table 3]

[0050] Thus, in certain embodiments, the antibody binds to the mature IL-18BP sequence of Table B1, for example, in the region excluding the signal peptide (underlined).

[0051] In certain embodiments, the antibody binds to a conformational epitope of the mature IL-18BP sequence of SEQ ID NO: 372 (mature human isoform A). In exemplary embodiments, an antibody of the disclosure binds to at least two residues selected from the group consisting of T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In exemplary embodiments, an antibody of the disclosure binds to residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In exemplary embodiments, the antibody binds to a V sequence with at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 347. H , and V of a sequence having at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 348 L In exemplary embodiments, the VHCDR1, VHCDR2, and VHCDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and the VLCDR1, VLCDR2, and VLCDR3 sequences comprise SEQ ID NOs: 250-252, respectively.

[0052] In certain embodiments, the antibody binds to an epitope comprising the IL-18-binding interface of mature IL-18B. Residues on IL-18BP that interact with IL-18 have been identified as follows: R61, Y69, S75, H79, T116, S119, and R131. In exemplary embodiments, an antibody of the present disclosure binds to residues S75, H79, T116, and S119, which are also recognized by IL-18. In exemplary embodiments, the antibody binds to the V region of a sequence with at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 347. H , and V of a sequence having at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identity to SEQ ID NO: 348 L In exemplary embodiments, the VHCDR1, VHCDR2, and VHCDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and the VLCDR1, VLCDR2, and VLCDR3 sequences comprise SEQ ID NOs: 250-252, respectively.

[0053] In certain embodiments, the antibody binds to a linear epitope of the mature IL-18BP sequence of SEQ ID NO: 372 (mature human isoform A). In certain embodiments, the antibody has orthologous specificity or orthologous cross-reactivity with respect to IL-18BP. For example, in certain embodiments, the antibody binds to human IL-18BP and cynomolgus monkey IL-18BP, but does not bind (specifically or substantially) to mouse IL-18BP. In some embodiments, the antibody binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

[0054] In some embodiments, the antibody is directed to human IL-18BP by measuring the binding affinity between IL-18 and IL-18BP (K DIn some examples, the antibody binds to human IL-18BP with a binding affinity of about 1 pM to about 10 pM to about 600 pM, or 65 pM, or about, at least about, or less than about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM, or optionally about 1 pM to about 600 pM, 1 pM to about 600 pM, 1 pM to about 650 pM, or 65 pM. pM to about 500pM, 1pM to about 400pM, 1pM to about 300pM, about 1pM to about 200pM, about 1pM to about 100pM, about 1pM to about 50pM, about 1pM to about 40pM, about 1pM to about 30pM, about 1pM to about 20pM, about 1 pM to about 10pM, about 1pM to about 5pM, about 5pM to about 600pM, about 5pM to about 500pM, about 5pM to about 400pM, about 5pM to about 300pM, about 5pM to about 200pM, about 5pM to about 100pM, about 5pM to about 50pM, about 5 pM to about 40 pM, about 5 pM to about 30 pM, about 5 pM to about 20 pM, about 5 pM to about 10 pM, about 10 pM to about 600 pM, about 10 pM to about 500 pM, about 10 pM to about 400 pM, about 10 pM to about 300 pM, about 10 pM to about 200 pM, about 10 pM to about 100 pM, about 10 pM to about 50 pM, about 10 pM to about 40 pM, about 10 pM to about 30 pM, about 10 pM to about 20 pM, or about 20 pM to about 600 pM, about 20 pM to about 500 pM, about 20 pM to about It binds with a binding affinity of about 400 pM, about 20 pM to about 300 pM, about 20 pM to about 200 pM, about 20 pM to about 100 pM, about 20 pM to about 50 pM, about 20 pM to about 40 pM, about 20 pM to about 30 pM, or about 30 pM to about 600 pM, about 30 pM to about 500 pM, about 30 pM to about 400 pM, about 30 pM to about 300 pM, about 30 pM to about 200 pM, about 30 pM to about 100 pM, about 30 pM to about 50 pM, or about 30 pM to about 40 pM. Dmay be determined by the biolayer interference (BLI) assay described herein. For example, binding kinetics measurements may be performed on a ForteBio (now Sartorius) Octet RED96e instrument by loading the mAb onto an anti-human constant domain (AHC) biosensor (ForteBio) in 10x kinetics buffer consisting of PBS containing 0.1% BSA and 0.02% Tween 20 for 90-120 seconds, achieving spectral shift values ​​of 0.8-1.2 nm. Binding may then be performed in the presence of a two-fold dilution series of hIL-18BP and allowed to proceed for 90-120 seconds. Dissociation may be measured over 300-1200 seconds. The dilution series may start at 100 nM for weaker variants or 10 nM for the most potent mAb.

[0055] In some embodiments, the antibody, or antigen-binding fragment thereof, is an IL-18BP antagonist. In some examples, the antibody, or antigen-binding fragment thereof, antagonizes the binding and / or signaling activity between IL-18BP and its ligand, IL-18. In some embodiments, the antibody, or antigen-binding fragment thereof, antagonizes or reduces the binding and / or signaling activity between IL-18BP and IL-18 by about or at least about 10-1000% (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000% or more), e.g., in a cell-based assay. In some examples, an antagonistic anti-IL-18BP antibody or antigen-binding fragment thereof blocks the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signal transduction, e.g., IL-18-mediated induction of IFN-γ, CXCL10, and TNFα. These functional activities may be measured by the assays disclosed herein. For example, an antibody may be incubated with IL-18BP (e.g., human IL-18BP) followed by the addition of IL-18 (e.g., recombinant human IL-18). The resulting solution may then be added to IL-18 reporter HEK293 cells. The cells respond to exogenously added IL-18 by expressing the NF-κB / AP-1-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene. The effect of the antibody may then be analyzed by its effect on the reporter cells compared with an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section of this specification. Another potential assay involves measuring the inhibition of IFNγ expression by anti-IL-18BP mAb in KG-1 cells. Briefly, IL-18BP may be blocked first using serial dilutions of the antibody, then IL-18 may be added to the mixture, and the mixture may be added to KG-1 cells and incubated.Secreted IFN-γ can then be measured according to standard means, such as ELISA, and the effect of the test antibody can be compared to that of an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section herein. Yet another possible assay involves incubating PBMCs with the test antibody, IL-12, and IL-18, and measuring IFN-γ and / or CCL2 by standard methods. The effect of the test antibody may be compared to that of an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section herein. A further assay involves incubation of NK cells with precomplexed hIL-18 / hIL-18BP, followed by the addition of IL-12, followed by the addition of serial dilutions of the test antibody. The effect of the test antibody is compared to that of an appropriate control, such as an isotype control antibody. Further details are disclosed in the Materials and Methods section herein.

[0056] Certain embodiments include a method of screening anti-IL-18BP antibodies for the ability to block or inhibit binding between IL-18 and IL-18BP, the method comprising: (a) determining the binding affinity of the antibody to (i) IL-18BP alone, and (ii) an IL-18-reduced fusion protein, the fusion protein comprising IL-18 fused to IL-18BP via a flexible linker (and optionally a protease cleavage site therebetween), the fusion the IL-18 portion of the protein is linked to the IL-18BP portion of the fusion protein to sterically block the IL-18 binding site of the IL-18BP portion of the fusion protein; (b) comparing the binding affinity of (i) with the binding affinity of (ii); and (c) identifying or selecting the antibody as having the ability to block or inhibit binding between IL-18 and IL-18BP if the binding affinity of (i) is significantly stronger than the binding affinity of (ii). Certain embodiments include (c) identifying or selecting the antibody as having the ability to block or inhibit binding between IL-18 and IL-18BP if the binding affinity of (i) is about or at least about 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, 200-fold, 300-fold, 400-fold, 500-fold, 600-fold, 700-fold, 800-fold, 900-fold, or 1000-fold or more stronger than the binding affinity of (ii). In certain embodiments, the IL-18 and IL-18BP are murine IL-18 and IL-18BP. In some embodiments, the IL-18 and IL-18BP are human IL-18 and IL-18BP. In some embodiments, the IL-18-reduced fusion protein comprises, from N- to C-terminus, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP. In certain embodiments, the IL-18-reduced fusion protein comprises an amino acid sequence that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1.

[0057] For illustrative purposes only, binding interactions (e.g., binding affinity) between any combination of IL-18BP, IL-18 (e.g., low IL-18), and / or anti-IL-18BP antibodies described herein, or binding / signaling between IL-18BP and IL-18, can be detected and quantified using a variety of routinely used methods, including Biacore® assays (e.g., coupled to a sensor chip using an appropriately tagged soluble reagent), FACS analysis using cells (either natural or recombinant) expressing IL-18BPr on their cell surface, immunoassays, fluorescent staining assays, ELISA assays, and microcalorimetry, e.g., ITC (isothermal titration calorimetry). Similarly, the functional properties of anti-IL-18BP antibodies can be evaluated using a variety of methods known to those skilled in the art, including affinity / binding assays (e.g., surface plasmon resonance, competitive inhibition assays); cytotoxicity assays, cell survival assays, cell proliferation or differentiation assays, and cancer cell and / or tumor growth inhibition using in vitro or in vivo models. Other assays may be used to test the ability of the antibodies described herein to modulate (e.g., inhibit) IL-18BP and / or IL-18-mediated responses. The antibodies described herein may be tested for in vitro and in vivo efficacy. Such assays may be performed using well-established protocols known to those skilled in the art (see, e.g., Current Protocols in Molecular Biology (Greene Publ. Assoc. Inc. & John Wiley & Sons, Inc., NY, NY); Current Protocols in Immunology (Edited by: John E. Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, Warren Strober 2001 John Wiley & Sons, NY, NY); or commercially available kits.

[0058] In certain embodiments, the Fc region of the antibody comprises, consists of, or consists essentially of an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or a hybrid and / or variant thereof. In certain embodiments, the Fc region comprises, consists of, or consists essentially of an Fc from human IgG1 or IgG4 (see, e.g., Allberse and Schuurman, Immunology. 105:9-19, 2002), or a fragment or variant thereof.

[0059] In certain embodiments, antibodies comprise variant or otherwise modified Fc regions, including those with altered properties or biological activity compared to a wild-type Fc region. Examples of modified Fc regions include regions having a mutated sequence, e.g., by substitution, insertion, deletion, or truncation of one or more amino acids relative to the wild-type sequence, hybrid Fc polypeptides composed of domains from different immunoglobulin classes / subclasses, Fc polypeptides with altered glycosylation / sialylation patterns, and Fc polypeptides that are modified or derivatized, e.g., by biotinylation (see, e.g., U.S. Patent Application Publication No. 2010 / 0209424), phosphorylation, sulfation, etc., or any combination of the foregoing. Such modifications can affect the binding properties of the Fc region to one or more specific FcRs (e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, FcγRIIIb, FcRn), its pharmacokinetic properties (e.g., stability or half-life, bioavailability, tissue distribution, volume of distribution, concentration, elimination rate constant, elimination rate, area under the curve (AUC), clearance, C max , T max , C min, wander mutations), its immunogenicity, its complement fixation or activation, and / or CDC / ADCC / ADCP-related activity of the Fc region compared to the corresponding wild-type Fc sequence of the antibody. Included are modified Fc regions of human and / or murine origin.

[0060] In certain embodiments, the antibody comprises a hybrid Fc region, e.g., an Fc region comprising a combination of Fc domains (e.g., hinge, CH2, CH3, CH4) from immunoglobulins of different species (e.g., human, mouse), different Ig classes, and / or different Ig subclasses. Also included are antibodies comprising derivatized or otherwise modified Fc regions. In certain aspects, the Fc region is modified, e.g., by phosphorylation, sulfation, acylation, glycosylation, methylation, farnesylation, acetylation, amidation, and the like, relative to a wild-type or native Fc region. In certain embodiments, the Fc region comprises a wild-type or native glycosylation pattern, or alternatively, it comprises increased glycosylation relative to the native form, decreased glycosylation relative to the native form, or it is completely deglycosylated. As an example of an altered Fc glycoform, reduced glycosylation of the Fc region reduces binding to the C1q region of the first complement component C1, decreases ADCC-related activity, and / or decreases CDC-related activity. Certain embodiments thus employ deglycosylated or aglycosylated Fc regions. For exemplary production of aglycosylated Fc regions, see, e.g., WO 2005 / 047337. Another example of an Fc region glycoform is generated by substituting a cysteine ​​residue at position Q295 according to the Kabat et al. numbering system (see, e.g., U.S. Patent Application No. 2010 / 0080794). Certain embodiments include Fc regions in which approximately 80-100% of the glycoproteins in the Fc region comprise a mature core carbohydrate structure lacking fucose (see, e.g., U.S. Patent Application No. 2010 / 0255013). Some embodiments include Fc regions that are optimized by substitutions or deletions to reduce the level of fucosylation, e.g., to increase affinity for FcγRI, FcγRIa, or FcγRIIIa, and / or to improve phagocytosis by FcγRIIa-expressing cells (see U.S. Patent Applications Nos. 2010 / 0249382 and 2007 / 0148170).

[0061] As another example of a modified Fc glycoform, the Fc region of the antibody may comprise oligomannose-type N-glycans, optionally having one or more of the following: increased ADCC effector activity, increased binding affinity for FcγRIIIA (and certain other FcRs), similar or increased binding specificity for the target of an IL-18BP polypeptide, similar or higher binding affinity for the target of an IL-18BP polypeptide, and / or similar or lower binding affinity for the mannose receptor, relative to a corresponding Fc region comprising complex-type N-glycans (see, e.g., U.S. Patent Application No. 2007 / 0092521 and U.S. Patent No. 7,700,321). As another example, increased affinity of the Fc region for FcγRs has been achieved using engineered glycoforms generated by expression of the antibody in engineered or variant cell lines (see, e.g., Umana et al., Nat Biotechnol. 17:176-180, 1999; Davies et al., Biotechnol Bioeng. 74:288-294, 2001; Shields et al., J Biol Chem. 277:26733-26740, 2002; Shinkawa et al., 2003, J Biol Chem. 278:3466-3473, 2003; and U.S. Patent Application No. 2007 / 0111281). Certain Fc region glycoforms contain an increased proportion of N-glycosidically linked complex glycans that do not have the 1-position of fucose attached to the 6-position of the N-acetylglucosamine at the reducing end of the glycan (see, e.g., U.S. Patent Application Publication No. 2010 / 0092997). Certain embodiments may include an IgG Fc region that is glycosylated with at least one galactose moiety linked to each terminal sialic acid moiety by an α-2,6 linkage, optionally wherein the Fc region has greater anti-inflammatory activity than the corresponding wild-type Fc region (see, e.g., U.S. Patent Application Publication No. 2008 / 0206246).Some of these and related altered glycosylation approaches, as described herein, have produced substantial enhancements in the ability of Fc regions to selectively bind to FcRs, such as FcγRIII, mediate ADCC, and modify other properties of the Fc region.

[0062] Particular variant, fragment, hybrid, or otherwise modified Fc regions of antibodies may have altered binding to one or more FcRs and / or corresponding changes in effector function compared to the corresponding wild-type Fc sequence (e.g., same species, same Ig class, same Ig subclass). For example, such Fc regions may have increased binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In other embodiments, variant, fragment, hybrid, or modified Fc regions may have decreased binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. Particular FcRs are described elsewhere herein.

[0063] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to increase binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors relative to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations to increase binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors relative to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to increase effector function. In some embodiments, at least one antibody comprises an Fc domain selected from human IgG1 and human IgG3 comprising one or more mutations to increase effector function.

[0064] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with enhanced effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG1 and human IgG3, with one or more mutations to increase effector function. In some embodiments, the antibody is a partial blocking antibody comprising an Fc domain with enhanced effector activity. In some embodiments, the partial blocking antibody comprises an Fc domain selected from human IgG1 and human IgG3, with one or more mutations to increase effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with enhanced effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG1 or human IgG3, with one or more mutations to increase effector function.

[0065] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to reduce binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations to reduce binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to reduce effector function. In some embodiments, the antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function.

[0066] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a partial blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the partial blocking antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG2 and human IgG4 comprising one or more mutations to reduce effector function.

[0067] Specific examples of Fc variants with altered (e.g., increased, decreased) effector function / FcR binding are described, for example, in U.S. Patent Nos. 5,624,821 and 7,425,619; U.S. Patent Application Nos. 2009 / 0017023, 2009 / 0010921, and 2010 / 0203046; and WO2000 / 42072 and WO2004 / 016750. Specific examples include human Fc regions with one or more substitutions at positions 298, 333, and / or 334, e.g., S298A, E333A, and / or K334A (based on the EU index numbering of Kabat et al.), which have been shown to increase binding to the activating receptor FcγRIIIa and decrease binding to the inhibitory receptor FcγRIIb. These mutations can be combined to produce double and triple mutant variants with further improvements in FcR binding. Particular embodiments include the S298A / E333A / K334A triple mutant, which increases binding to FcγRIIIa, decreases binding to FcγRIIb, and increases ADCC (see, e.g., Shields et al., J. Biol. Chem. 276:6591-6604, 2001; and Presta et al., Biochem. Soc. Trans. 30:487-490, 2002). See also Umana et al. (supra); and engineered Fc glycoforms with increased FcR binding, as disclosed in U.S. Pat. No. 7,662,925. Some embodiments include an Fc region comprising one or more substitutions based on the EU index of Kabat et al. selected from 434S, 252Y / 428L, 252Y / 434S, and 428L / 434S (see U.S. Patent Application Nos. 2009 / 0163699 and 20060173170). Some embodiments include an Fc region comprising one or more substitutions based on the EU index of Kabat et al. selected from L234A, L235A, and G237A (see U.S. Patent Application No. 17 / 779,425). Some embodiments include an Fc region comprising substitutions at L234A and L235A based on the EU index of Kabat et al.Some embodiments include an Fc region comprising substitutions at L234A and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising substitutions at L235A and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising substitutions at L234A, L235A, and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising substitutions at L234A, L235A, and G237A based on the EU index of Kabat et al. Some embodiments include an Fc region comprising one or more substitutions selected from M252Y, S254T, and T256E based on the EU index of Kabat et al. Some embodiments include an Fc region comprising one or more substitutions selected from M428L and N434S based on the EU index of Kabat et al. In some embodiments, the Fc substitutions disclosed herein are substitutions into an Fc domain selected from human IgG1, IgG2, IgG3, or IgG4. In some embodiments, the Fc substitutions disclosed herein are substitutions into a human IgG1 Fc domain. In some embodiments, the Fc substitutions disclosed herein are substitutions into a human IgG2 Fc domain. In some embodiments, the Fc substitutions disclosed herein are substitutions into a human IgG3 Fc domain. In some embodiments, the Fc substitutions disclosed herein are substitutions into a human IgG4 Fc domain. In some embodiments, the antibodies of the present disclosure comprise an Fc substitution disclosed herein and a V that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to each sequence from one of the named antibodies of Table A2 (e.g., SA04a). H Array and V L The antibody comprises a sequence, in which case the antibody comprises the CDRs of one of the named antibodies (e.g., SA04a) listed in Table A1.

[0068] Certain variant hybrids or modified Fc regions may have altered solubility compared to the corresponding wild-type Fc sequence. In certain embodiments, such Fc regions may have increased solubility compared to the corresponding wild-type Fc sequence. In other embodiments, variant hybrids or modified Fc regions may have decreased solubility compared to the corresponding wild-type Fc sequence. Solubility can be measured, for example, in vitro (e.g., under physiological conditions) according to routine techniques in the art. Exemplary solubility measurements are described elsewhere herein.

[0069] The variant Fc region can also have one or more mutant hinge regions, for example, as described in U.S. Patent Application Publication No. 2003 / 0118592. For example, one or more cysteines in the hinge region can be deleted or substituted with different amino acids. The mutant hinge region can contain no cysteine ​​residues, or it can contain one, two, or three fewer cysteine ​​residues than the corresponding wild-type hinge region. In some embodiments, Fc regions with this type of mutant hinge region exhibit reduced dimerization ability compared to the wild-type Ig hinge region.

[0070] In some embodiments, the antibody or antigen-binding fragment thereof may be conjugated to one or more cytotoxic or chemotherapeutic agents. In some embodiments, the antibodies disclosed herein are conjugated or operably linked to a radioisotope to form a radioconjugate and / or a macrocyclic chelator useful for complexing radioactive metal ions. The antibodies used in any of the compositions, methods, and / or kits described herein can be combined with one or more of the additional agents described herein.

[0071] Methods of Use and Pharmaceutical Compositions Certain embodiments relate to methods of treating, ameliorating symptoms, and / or slowing the progression of a disease or condition in a subject in need thereof, comprising administering to the subject an antibody that binds to IL-18BP, or a pharmaceutical composition comprising the same, as described herein. Also included are methods of stimulating an immune response, e.g., an IL-18-mediated immune response, in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein. In some examples, the antibody antagonizes the binding / signaling activity between IL-18BP and its ligand, IL-18, thereby increasing IL-18-mediated signaling or activity (e.g., increased induction of IFN-gamma, CXCL10, and / or TNFα). In some embodiments, the disease or condition is cancer or tumor, or an infectious disease, as described above. In some embodiments, the disease is any disease in which activation of the immune system may be beneficial.

[0072] In some embodiments, the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, such as a lymphoproliferative disorder, a myeloproliferative disorder, proliferative enterocolitis, proliferative diabetic retinopathy, or a proliferative renal disease, as described above. In some examples, the cancer or tumor expresses or overexpresses IL-18BP, IL-18, or both. In some examples, the proliferative disease or disorder is associated with increased expression of IL-18BP, IL-18, or both. In some examples, the cancer is a primary cancer. In some examples, the cancer is a metastatic cancer. Accordingly, certain embodiments include a method of treating, reducing the severity of, or preventing cancer in a patient in need thereof, comprising administering to the patient a composition described herein, wherein the antibody is an IL-18BP antagonist, thereby treating, reducing the severity of, or preventing cancer.

[0073] Examples of cancer include, but are not limited to, bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (HCC), and hepatocellular carcinoma (HCC). Cancers include, for example, alveolar carcinoma (hepatocellular carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer. In certain embodiments, the cancer is metastatic cancer, e.g., that has metastasized to bone.

[0074] Also provided are antibodies, antigen-binding fragments thereof, or pharmaceutical compositions of the present disclosure for use as pharmaceuticals. The antibodies, fragments thereof, or pharmaceutical compositions of the present disclosure may be for use in any of the therapeutic methods disclosed herein. In certain embodiments, the antibodies, fragments thereof, or pharmaceutical compositions of the present disclosure may be for use in a method of treating, ameliorating the symptoms of, and / or slowing the progression of any of the diseases or disorders disclosed herein, such as cancer, tumors, or other proliferative diseases or disorders, such as lymphoproliferative disorders, myeloproliferative disorders, proliferative enterocolitis, proliferative diabetic retinopathy, or proliferative kidney diseases.

[0075] Certain embodiments include combination therapies, e.g., the combination therapies include administering a pharmaceutical composition described herein (comprising an anti-IL-18BP antibody) in combination with one or more additional therapeutic agents, e.g., an immunostimulatory agent, an immune checkpoint modulator, and / or a chemotherapeutic agent. In some embodiments, the additional therapeutic agent comprises IL-18, including human IL-18 (or a functional variant or fragment thereof).

[0076] In some embodiments, the additional therapeutic agent comprises an immune checkpoint modulator. Specific examples of immune checkpoint modulators include "antagonists" of one or more inhibitory immune checkpoint molecules and "agonists" of one or more stimulatory immune checkpoint molecules. Generally, immune checkpoint molecules are components of the immune system that either upregulate (costimulatory molecules) or downregulate (decreasing) signals. Targeting immune checkpoint molecules has therapeutic potential in cancer, as cancer cells can disrupt the natural function of immune checkpoint molecules (see, e.g., Sharma and Allison, Science. 348:56-61, 2015; Topalian et al., Cancer Cell. 27:450-461, 2015; Pardoll, Nature Reviews Cancer. 12:252-264, 2012). In some embodiments, an immune checkpoint modulator (e.g., antagonist, agonist) "binds" or "specifically binds" to one or more immune checkpoint molecules, as described herein.

[0077] In certain embodiments, the immune checkpoint modulator is a polypeptide or peptide. The terms "peptide" and "polypeptide" are used interchangeably herein, although in certain instances, the term "peptide" can refer to a short polypeptide, e.g., a polypeptide consisting of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 amino acids (including all integers and ranges therebetween, e.g., 5-10, 8-12, 10-15). Polypeptides and peptides may be composed of naturally occurring and / or non-naturally occurring amino acids, as described herein.

[0078] Antibodies are also included as polypeptides. Thus, in some embodiments, an immune checkpoint modulating polypeptide agent is an "antibody," as described herein.

[0079] In some embodiments, the agent is or includes a "ligand," e.g., a native ligand of an immune checkpoint molecule. "Ligand" generally refers to a substance or molecule that forms a complex with a target molecule (e.g., a biomolecule) to achieve a biological purpose, and includes "protein ligands." Protein ligands generally generate a signal by binding to a site on a target molecule or target protein. Thus, certain agents are protein ligands that naturally bind to an immune checkpoint molecule and generate a signal. Also included are "modified ligands." Modified ligands are, for example, pharmacokinetic modifiers, e.g., protein ligands fused to an Fc region derived from an immunoglobulin.

[0080] In some embodiments, the agent is a "small molecule," which refers to an organic compound of synthetic or biological origin (biomolecule), but which is often not a polymer. Organic compounds refer to a large class of chemical compounds whose molecules contain carbon, typically excluding those that contain only carbonates, simple oxides of carbon, or cyanides. "Biomolecules" generally refer to organic molecules produced by living organisms, including large macromolecules (biopolymers), such as peptides, polysaccharides, and nucleic acids, as well as small molecules, such as primary and secondary metabolites, lipids, phospholipids, glycolipids, sterols, glycerolipids, vitamins, and hormones. "Polymer" generally refers to a large molecule or macromolecule composed of repeating structural units, typically linked by covalent bonds.

[0081] In some embodiments, the immune checkpoint modulator is an antagonist or inhibitor of one or more inhibitory immune checkpoint molecules, including Programmed Death-Ligand 1 (PD-L1), Programmed Death-Ligand 2 (PD-L2), Programmed Death 1 (PD-1), Cytotoxic T-Lymphocyte-Associated Protein 4 (CTLA-4), Indoleamine 2,3-Dioxygenase (IDO), Tryptophan 2,3-Dioxygenase (TDO), T-cell Immunoglobulin Domain and Mucin Domain 3 (TIM-3), Lymphocyte Activation Gene-3 (LAG-3), V-domain Ig suppressor of T cell activation (VISTA), B and T Lymphocyte Attenuator (BTLA), CD160, and T-cell immunoreceptor with Ig and ITIM domains (TIGIT).

[0082] In certain embodiments, the agent is a PD-1 (receptor) antagonist or inhibitor, the targeting of which has been shown to restore immune function in the tumor environment (see, e.g., Phillips et al., Int Immunol. 27:39-46, 2015). PD-1 is a cell surface receptor belonging to the immunoglobulin superfamily and expressed on T cells and pro-B cells. PD-1 interacts with two ligands, PD-L1 and PD-L2. PD-1 functions as an inhibitory immune checkpoint molecule, for example, by reducing or preventing T cell activation, which in turn reduces autoimmunity and promotes self-tolerance. The inhibitory effect of PD-1 is achieved, at least in part, through a dual mechanism: promoting apoptosis of antigen-specific T cells in lymph nodes while reducing apoptosis of regulatory T cells (suppressor T cells). Some examples of PD-1 antagonists or inhibitors include antibodies or small molecules that specifically bind to PD-1 and reduce one or more of its immunosuppressive activities, such as downstream signaling or interaction with PD-L1. Specific examples of PD-1 antagonists or inhibitors include the antibodies nivolumab, pembrolizumab, PDR001, MK-3475, AMP-224, AMP-514, and pidilizumab, and antigen-binding fragments thereof (see, e.g., U.S. Patent Nos. 8,008,449, 8,993,731, 9,073,994, 9,084,776, 9,102,727, 9,102,728, 9,181,342, 9,217,034, 9,387,247, 9,492,539, 9,492,540, and U.S. Patent Application Nos. 2012 / 0039906 and 2015 / 0203579).

[0083] In some embodiments, the agent is a PD-L1 antagonist or inhibitor. As mentioned above, PD-L1 is one of the natural ligands of the PD-1 receptor. Common examples of PD-L1 antagonists or inhibitors include antibodies or small molecules that specifically bind to PD-L1 and reduce one or more of its immunosuppressive activities, such as binding to the PD-1 receptor. Specific examples of PD-L1 antagonists include the antibodies atezolizumab (MPDL3280A), avelumab (MSB0010718C), and durvalumab (MEDI4736), and antigen-binding fragments thereof (see, e.g., U.S. Patent Nos. 9,102,725, 9,393,301, 9,402,899, and 9,439,962).

[0084] In some embodiments, the agent is a PD-L2 antagonist or inhibitor. As discussed above, PD-L2 is one of the natural ligands of the PD-1 receptor. Common examples of PD-L2 antagonists or inhibitors include antibodies or small molecules that specifically bind to PD-L2 and reduce one or more of its immunosuppressive activities, such as binding to the PD-1 receptor.

[0085] In some embodiments, the agent is a CTLA-4 antagonist or inhibitor. CTLA4 or CTLA-4 (cytotoxic T-lymphocyte-associated protein 4), also known as CD152 (cluster of differentiation 152), is a protein receptor that functions as an inhibitory immune checkpoint molecule by sending an inhibitory signal to T cells when it binds to CD80 or CD86, for example, on the surface of antigen-presenting cells. Common examples of CTLA-4 antagonists or inhibitors include antibodies or small molecules that specifically bind to CTLA-4. Specific examples include the antibodies ipilimumab and tremelimumab, and antigen-binding fragments thereof. At least part of the activity of ipilimumab is believed to be mediated by antibody-dependent cell-mediated cytotoxicity (ADCC) killing of CTLA-4-expressing suppressor Tregs.

[0086] In some embodiments, the agent is an IDO antagonist or inhibitor, or a TDO antagonist or inhibitor. IDO and TDO are tryptophan catabolic enzymes with immunosuppressive properties. For example, IDO is known to suppress T cells and NK cells, generate and activate Tregs and myeloid-derived suppressor cells, and promote tumor angiogenesis. Common examples of IDO and TDO antagonists or inhibitors include antibodies or small molecules that specifically bind to IDO or TDO (see, e.g., Platten et al., Front Immunol. 5:673, 2014) and reduce or inhibit one or more immunosuppressive activities. Specific examples of IDO antagonists or inhibitors include indoximod (NLG-8189), 1-methyl-tryptophan (1MT), β-carboline (norharman, 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat (see, e.g., Sheridan, Nature Biotechnology. 33: 321-322, 2015). Specific examples of TDO antagonists or inhibitors include 680C91 and LM10 (see, e.g., Pilotte et al., PNAS USA. 109: 2497-2502, 2012).

[0087] In some embodiments, the agent is a TIM-3 antagonist or inhibitor. T-cell immunoglobulin domain and mucin domain 3 (TIM-3) is expressed on activated human CD4+ T cells and regulates Th1 and Th17 cytokines. TIM-3 also acts as a negative regulator of Th1 / Tc1 function by inducing cell death upon interaction with its ligand, galectin-9. TIM-3 contributes to a suppressive tumor microenvironment, and its overexpression is associated with poor prognosis in various cancers (see, e.g., Li et al., Acta Oncol. 54:1706-13, 2015). Common examples of TIM-3 antagonists or inhibitors include antibodies or small molecules that specifically bind to TIM-3 and reduce or inhibit one or more of its immunosuppressive activities.

[0088] In some embodiments, the agent is an antagonist or inhibitor of LAG-3. Lymphocyte Activation Gene-3 (LAG-3) is expressed on activated T cells, natural killer cells, B cells, and plasmacytoid dendritic cells. Like CTLA-4 and PD-1, LAG-3 negatively regulates T cell proliferation, activation, and homeostasis (see, e.g., Workman and Vignali, European Journal of Immun. 33:970-9, 2003; and Workman et al., Journal of Immun. 172:5450-5, 2004) and has been reported to play an important role in the suppressive function of Tregs (see, e.g., Huang et al., Immunity. 21:503-13, 2004). LAG3 also maintains CD8+ T cells in a tolerogenic state and associates with PD-1 to maintain the exhaustion of CD8 T cells. General examples of LAG-3 antagonists or inhibitors include antibodies or small molecules that specifically bind to LAG-3 and inhibit one or more of its immunosuppressive activities. Specific examples include the antibody BMS-986016 and antigen-binding fragments thereof.

[0089] In some embodiments, the agent is a VISTA antagonist or inhibitor. V-domain Ig suppressor of T cell activation (VISTA) is an inhibitory immune checkpoint regulator that is expressed primarily on hematopoietic cells, suppresses T cell activation, induces Foxp3 expression, and is highly expressed in the tumor microenvironment. VISTA suppresses anti-tumor T cell responses in the tumor microenvironment (see, e.g., Lines et al., Cancer Res. 74:1924-32, 2014). Common examples of VISTA antagonists or inhibitors include antibodies or small molecules that specifically bind to VISTA and reduce one or more of its immunosuppressive activities.

[0090] In some embodiments, the agent is a BTLA antagonist or inhibitor. Expression of B- and T-lymphocyte attenuator (BTLA; CD272) is induced during T cell activation and inhibits T cells through interaction with tumor necrosis family receptors (TNF-R) and the B7 family of cell surface receptors. BTLA is a ligand for tumor necrosis factor (receptor) superfamily, member 14 (TNFRSF14) and is also known as herpes virus entry mediator (HVEM). The BTLA-HVEM complex negatively regulates T cell immune responses, for example, by inhibiting the function of human CD8+ cancer-specific T cells (see, e.g., Derre et al., J Clin Invest 120:157-67, 2009). Common examples of BTLA antagonists or inhibitors include antibodies or small molecules that specifically bind to BTLA-4 and reduce one or more of its immunosuppressive activities.

[0091] In some embodiments, the agent is an antagonist or inhibitor of HVEM, e.g., an antagonist or inhibitor that specifically binds to HVEM and interferes with its interaction with BTLA or CD 160. Common examples of HVEM antagonists or inhibitors include antibodies or small molecules that specifically bind to HVEM and optionally reduce HVEM / BTLA and / or HVEM / CD 160 interactions, thereby reducing one or more of the immunosuppressive activities of HVEM.

[0092] In some embodiments, the agent is a CD160 antagonist or inhibitor, e.g., an antagonist or inhibitor that specifically binds to CD160 and interferes with its interaction with HVEM. Common examples of CD160 antagonists or inhibitors include antibodies or small molecules that specifically bind to CD160 and optionally reduce the CD160 / HVEM interaction, thereby reducing or inhibiting one or more of its immunosuppressive activities.

[0093] In some embodiments, the agent is a TIGIT antagonist or inhibitor. T cell Ig and ITIM domain (TIGIT) is a co-inhibitory receptor present on the surface of various lymphoid cells that suppresses anti-tumor immunity, for example, via Tregs (Kurtulus et al., J Clin Invest. 125:4053-4062, 2015). Common examples of TIGIT antagonists or inhibitors include antibodies or small molecules that specifically bind to TIGIT and reduce one or more of its immunosuppressive activities (see, e.g., Johnston et al., Cancer Cell. 26:923-37, 2014).

[0094] In certain embodiments, the immune checkpoint modulator is an agonist of one or more stimulatory immune checkpoint molecules, examples of which include OX40, CD40, Glucocorticoid-Induced TNFR Family Related Gene (GITR), CD137 (4-1BB), CD27, CD28, CD226, and Herpes Virus Entry Mediator (HVEM).

[0095] In some embodiments, the agent is an OX40 agonist. OX40 (CD134) promotes the proliferation of effector and memory T cells and suppresses the differentiation and activity of T regulatory cells (see, e.g., Croft et al., Immunol Rev. 229:173-91, 2009). Its ligand is OX40L (CD252). OX40 signaling influences both T cell activation and survival, and therefore plays an important role in initiating anti-tumor immune responses in lymph nodes and maintaining anti-tumor immune responses in the tumor microenvironment. Common examples of OX40 agonists include antibodies, small molecules, or ligands that specifically bind to OX40 and increase one or more of its immunostimulatory activities. Specific examples include OX86, OX-40L, Fc-OX40L, GSK3174998, MEDI0562 (humanized OX40 agonist), MEDI6469 (murine OX4 agonist), and MEDI6383 (OX40 agonist), and antigen-binding fragments thereof.

[0096] In some embodiments, the agent is a CD40 agonist. CD40 is expressed on antigen-presenting cells (APCs) and some malignant tumors. Its ligand is CD40L (CD154). On APCs, ligation results in upregulation of costimulatory molecules, potentially eliminating the need for T cell help in antitumor immune responses. CD40 agonist therapy plays an important role in APC maturation and their migration from tumors to lymph nodes, resulting in improved antigen presentation and T cell activation. Anti-CD40 agonist antibodies have induced robust responses and durable anticancer immunity in animal models, effects mediated, at least in part, by cytotoxic T cells (see, e.g., Johnson et al., Clin Cancer Res. 21:1321-1328, 2015; and Vonderheide and Glennie, Clin Cancer Res. 19:1035-43, 2013). Common examples of CD40 agonists include antibodies, small molecules, or ligands that specifically bind to CD40 and increase one or more of its immunostimulatory activities. Specific examples include sotigalilmab, CP-870,893, dacetuzumab, Chi Lob 7 / 4, ADC-1013, CD40L, rhCD40L, and antigen-binding fragments thereof.

[0097] In some embodiments, the agent is a GITR agonist. Glucocorticoid-Induced TNFR family Related gene (GITR) increases T cell expansion, inhibits Treg suppressive activity, and prolongs the survival of T effector cells. GITR agonists have been shown to promote anti-tumor responses by disrupting the stability of the Treg lineage (see, e.g., Schaer et al., Cancer Immunol Res. 1:320-31, 2013). These diverse mechanisms indicate that GITR plays an important role in initiating immune responses in lymph nodes and maintaining immune responses in tumor tissues. Its ligand is GITRL. Common examples of GITR agonists include antibodies, small molecules, or ligands that specifically bind to GITR and increase one or more of its immunostimulatory activities. Specific examples include GITRL, INCAGN01876, DTA-1, MEDI1873, and antigen-binding fragments thereof.

[0098] In some embodiments, the agent is a CD137 agonist. CD137 (4-1BB) is a member of the tumor necrosis factor (TNF) receptor family, and cross-linking of CD137 enhances T cell proliferation, IL-2 secretion, survival, and cytolytic activity. CD137-mediated signaling also protects T cells, such as CD8+ T cells, from activation-induced cell death. Common examples of CD137 agonists include antibodies, small molecules, or ligands that specifically bind to CD137 and increase one or more of its immunostimulatory activities. Specific examples include CD137 (or 4-1BB) ligands (see, e.g., Shao and Schwarz, J. Leukoc. Biol. 89:21-9, 2011), and utomirumab, an antibody comprising an antigen-binding fragment thereof.

[0099] In some embodiments, the agent is a CD27 agonist. Stimulation of CD27 increases the antigen-specific expansion of naive T cells and contributes to the long-term maintenance of T cell memory and T cell immunity. Its ligand is CD70. Targeting human CD27 with agonistic antibodies stimulates T cell activation and antitumor immunity (see, e.g., Thomas et al., Oncoimmunology. 2014;3:e27255.doi:10.4161 / onci.27255; and He et al., J Immunol. 191:4174-83, 2013). General examples of CD27 agonists include antibodies, small molecules, or ligands that specifically bind to CD27 and increase one or more of its immunostimulatory activities. Specific examples include the antibodies valilumab and CDX-1127 (1F5), which contain CD70 and antigen-binding fragments thereof.

[0100] In some embodiments, the agent is a CD28 agonist. CD28 is constitutively expressed on CD4+ T cells and a subset of CD8+ T cells. Its ligands include CD80 and CD86, and stimulation of these ligands increases T cell proliferation. Common examples of CD28 agonists include antibodies, small molecules, or ligands that specifically bind to CD28 and increase one or more of its immunostimulatory activities. Specific examples include CD80, CD86, the antibody TAB08, and antigen-binding fragments thereof.

[0101] In some embodiments, the agent is a CD226 agonist. CD226 shares a ligand with TIGIT and is a stimulatory receptor opposite to TIGIT. CD226 engagement enhances T cell activation (see, for example, Kurtulus et al., J Clin Invest. 125:4053-4062, 2015; Bottino et al., J Exp Med. 1984:557-567, 2003; and Tahara-Hanaoka et al., Int Immunol. 16:533-538, 2004). Common examples of CD226 agonists include antibodies, small molecules, or ligands (e.g., CD112, CD155) that specifically bind to CD226 and increase one or more of its immunostimulatory activities.

[0102] In some embodiments, the agent is an HVEM agonist. Herpesvirus entry mediator (HVEM), also known as tumor necrosis factor receptor superfamily member 14 (TNFRSF14), is a human cell surface receptor of the TNF receptor superfamily. HVEM is present on a variety of cells, including T cells, APCs, and other immune cells. Unlike other receptors, HVEM is expressed at high levels on resting T cells and is downregulated upon activation. HVEM signaling has been shown to play an important role in the early stages of T cell activation and during the expansion of tumor-specific lymphocyte populations in lymph nodes. Common examples of HVEM agonists include antibodies, small molecules, or ligands that specifically bind to HVEM and increase one or more of its immunostimulatory activities.

[0103] In some embodiments, the additional therapeutic agent comprises a chemotherapeutic agent, e.g., a small molecule chemotherapeutic agent, non-limiting examples of which include alkylating agents, antimetabolites, cytotoxic antibiotics, topoisomerase inhibitors (type 1 or type II), and anti-microtubule agents, among others.

[0104] In certain embodiments, the methods and compositions described herein are sufficient to cause a statistically significant decrease in the amount of viable tumor, e.g., at least a 10%, 20%, 30%, 40%, 50% or greater decrease in tumor mass, or tumor regression as indicated by an altered (e.g., statistically significantly decreased) scan dimension. In some embodiments, the methods and compositions described herein reduce the rate of cancer growth (e.g., in vivo or in vitro, including cancer cells isolated from a biopsy or other sample and grown in vitro) by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to an untreated control. In some examples, the methods and compositions described herein reduce cancer cell initiation, migration, adhesion, invasiveness, and / or metastasis by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to untreated controls. In some examples, the methods and compositions described herein reduce angiogenesis in the tumor environment by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to untreated controls.

[0105] In certain embodiments, the disease or condition is myelodysplastic syndrome (MDS) (see, e.g., Wang et al., Blood. 140 (Supplement 1):12297, 2022). For example, in response, methods of antagonizing IL-18BP are a viable approach. MDS refers to a group of cancers in which immature blood cells in the bone marrow fail to mature and, as a result, fail to develop into healthy blood cells. Accordingly, certain embodiments include methods of treating, reducing the severity of, or preventing MDS in a patient in need thereof, comprising administering to the patient a composition described herein, wherein the antibody is an IL-18BP antagonist, thereby treating, reducing the severity of, or preventing MDS.

[0106] In some embodiments, the disease or condition is an infectious disease. For example, in certain embodiments, the infectious disease is selected from a viral (see, e.g., Vecchie et al., J Cell Physiol. 236(3):1638-1657, 2021), bacterial (see, e.g., Kinoshita et al., Ann Surg. 240(2):313-20, 2004), fungal (e.g., yeast), and protozoan infection. Accordingly, some embodiments include a method of treating, reducing the severity of, or preventing an infectious disease in a patient in need thereof, the method comprising administering to the patient a composition described herein, wherein the antibody is an IL-18BP antagonist, thereby treating, reducing the severity of, or preventing the infectious disease.

[0107] In certain embodiments, the methods and compositions described herein are sufficient to result in stable disease, hi certain embodiments, the methods and compositions described herein are sufficient to result in a clinically relevant reduction in symptoms of a particular disease indication known to a skilled clinician.

[0108] For in vivo use, certain embodiments include pharmaceutical compositions comprising an antibody described herein and a pharmaceutically acceptable carrier. To prepare a therapeutic or pharmaceutical composition, an effective or desired amount of one or more agents is mixed with any pharmaceutical carrier or excipient known to those of skill in the art to be appropriate for the particular agent and / or mode of administration. Pharmaceutical carriers can be liquid, semi-liquid, or solid. Solutions or suspensions used for parenteral, intradermal, intraocular, subcutaneous, direct instillation into the bladder, or topical application can contain, for example, sterile diluents (such as water), saline (e.g., phosphate-buffered saline; PBS), fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents (such as benzyl alcohol and methylparabens); antioxidants (such as ascorbic acid and sodium bisulfite) and chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); buffers (such as acetates, citrates, and phosphates). If administered intravenously (e.g., by IV infusion), suitable carriers include saline or phosphate buffered saline (PBS), as well as solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof.

[0109] Administration of the agents described herein, in pure form or in suitable therapeutic or pharmaceutical compositions, can be via any of the accepted modes of administration for agents that perform similar functions. Therapeutic or pharmaceutical compositions can be prepared by combining the agent-containing composition with a suitable physiologically acceptable carrier, diluent, or excipient, and can be formulated into preparations in solid, semi-solid, liquid, or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. Other pharmaceutically active ingredients (including other small molecules described elsewhere herein) and / or suitable excipients, such as salts, buffers, and stabilizers, can also be present in the composition, but need not be.

[0110] Administration can be achieved by a variety of different routes, including oral, parenteral, nasal, intravenous, ocular, intradermal, intramuscular, subcutaneous, placement in the bladder, inhalation, sublingual, buccal, rectal, vaginal, or topical. The preferred mode of administration depends on the nature of the condition being treated or prevented. Certain embodiments include administration by IV infusion.

[0111] Carriers can include, for example, pharmaceutically or physiologically acceptable carriers, excipients, or stabilizers that are nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. Often, the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include buffers, such as phosphate, citrate, other organic acids, and the like; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine, histidine, and / or lysine, and the like; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or non-ionic surfactants, such as polysorbate 20 (TWEEN™), polyethylene glycol (PEG), poloxamer (PLURONICS™), and the like.

[0112] In some embodiments, one or more agents can be encapsulated in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacylate) microcapsules, respectively), in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980). The particles or liposomes may further comprise other therapeutic or diagnostic agents.

[0113] The exact dosage and duration of treatment depend on the disease to be treated, and in some cases may be empirically determined.Controlled clinical trials may also be carried out.Dosage may also vary depending on the severity of the condition to be alleviated.Pharmaceutical compositions are generally formulated and administered to exert therapeutically useful effects while minimizing undesirable side effects.The composition may be administered once, or may be divided into multiple smaller doses that are administered at regular intervals.For any particular subject, the specific dosage regimen may be adjusted over time according to individual needs.

[0114] Thus, typical routes of administration of these and related therapeutic or pharmaceutical compositions include, but are not limited to, oral, parenteral, nasal, intravenous, ocular, intradermal, intramuscular, subcutaneous, placement in the bladder, transdermal, inhalation, sublingual, buccal, rectal, vaginal, and topical. As used herein, the term "parenteral" includes subcutaneous injection, intravenous, injection into the bladder, intramuscular, intrasternal injection, or infusion techniques. Therapeutic or pharmaceutical compositions according to certain embodiments of the present disclosure are formulated so that the active ingredients contained therein are bioavailable upon administration of the composition to a subject or patient. The composition administered to a subject or patient may take the form of one or more dosage units, where, for example, a tablet may be a single dosage unit and a container of the agents described herein in aerosol form may hold multiple dosage units. Actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 23rd Edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered typically contains a therapeutically effective amount of an agent described herein for treatment of the disease or condition of interest.

[0115] Therapeutic or pharmaceutical compositions can be in solid or liquid form.In some embodiments, the carrier is particulate, and the composition is, for example, in tablet or powder form.The carrier can be liquid, and the composition is, for example, oral oil, injectable liquid, or aerosol, which is useful, for example, in inhalation administration.When intended for oral administration, pharmaceutical compositions are preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension, and gel form are included in the form considered herein as either solid or liquid.Certain embodiments include sterile injection solutions.

[0116] As a solid composition for oral administration, the pharmaceutical composition may be formulated into a powder, granules, gel, compressed tablet, pill, capsule, chewing gum, wafer, or the like. Such solid compositions typically contain one or more inert diluents or edible carriers. One or more of the following may also be present: binders such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, tragacanth, or gelatin; excipients such as starch, lactose, or dextrin; disintegrants such as alginic acid, sodium alginate, Primogel, corn starch, and the like; lubricants such as magnesium stearate or Stereotex; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; flavorings such as peppermint, methyl salicylate, or orange flavor; and coloring agents. When the pharmaceutical composition is in the form of a capsule, e.g., a gelatin capsule, it may contain, in addition to the above materials, a liquid carrier such as polyethylene glycol or oil.

[0117] Therapeutic or pharmaceutical compositions may be in the form of a liquid, such as an elixir, syrup, solution, gel, emulsion, or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, preferred compositions contain, in addition to the compound, one or more of a sweetener, a preservative, a dye / colorant, and a flavor enhancer. In compositions intended to be administered by injection, one or more of a surfactant, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffer, a stabilizer, and an isotonic agent may be included.

[0118] Liquid therapeutic or pharmaceutical compositions, whether in solution, suspension, or other similar form, may contain one or more of the following adjuvants: sterile diluents, such as water for injection, saline, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils, such as synthetic mono- or diglycerides, polyethylene glycols, glycerin, propylene glycol, or other solvents that serve as solvents or suspending media; antibacterial agents, such as benzyl alcohol or methylparabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, phosphates, and agents for adjusting tonicity, such as sodium chloride or glucose. Parenteral preparations can be enclosed in ampoules, disposable syringes, or multiple-dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. Pharmaceutical compositions for injection are preferably sterilized.

[0119] Liquid therapeutic or pharmaceutical compositions intended for either parenteral, ocular, or oral administration should contain an amount of agent such that an appropriate dosage will be obtained. Typically, this amount is at least 0.01% of the agent of interest in the composition. When intended for oral administration, this amount can vary to be between 0.1 and about 70% by weight of the composition. Certain oral therapeutic or pharmaceutical compositions contain between about 4% and about 75% of the agent of interest. In certain embodiments, therapeutic or pharmaceutical compositions and preparations are prepared so that a parenteral dosage unit contains between 0.01 and 10% by weight of the agent of interest before dilution.

[0120] Therapeutic or pharmaceutical compositions may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment, or gel base. The base may comprise, for example, one or more of the following: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickeners may also be present in therapeutic or pharmaceutical compositions for topical administration. If intended for transdermal administration, the composition may comprise a transdermal patch or iontophoresis device.

[0121] Therapeutic or pharmaceutical compositions may be intended for rectal administration, for example, in the form of suppositories that dissolve in the rectum and release the drug.Compositions for rectal administration may contain an oily base as a suitable non-irritating excipient.Such bases include, but are not limited to, lanolin, cocoa butter, and polyethylene glycol.

[0122] Therapeutic or pharmaceutical compositions may contain various materials that modify the physical form of a solid or liquid dosage unit. For example, the composition may contain a material that forms a coating shell around the active ingredient. The material that forms the coating shell is typically inert and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be encapsulated in a gelatin capsule. Therapeutic or pharmaceutical compositions in solid or liquid form may contain a component that binds to the drug and thereby assists in the delivery of the compound. Suitable components that can act in this capacity include monoclonal or polyclonal antibodies, one or more proteins, or liposomes.

[0123] Therapeutic or pharmaceutical compositions may consist essentially of dosage units that can be administered as an aerosol. The term "aerosol" is used to refer to a variety of systems, ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by liquefied or compressed gas or by an appropriate pump system that dispenses the active ingredient. The aerosol may be delivered in a single-phase, two-phase, or three-phase system to deliver the active ingredient. Aerosol delivery includes the necessary containers, activators, valves, subcontainers, and the like, which may together form a kit. One skilled in the art can determine a preferred aerosol without undue experimentation.

[0124] The compositions described herein may be prepared with carriers that protect the agent against rapid elimination from the body, such as slow release formulations or coatings. Such carriers include controlled release formulations, including, but not limited to, implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid, and others known to those skilled in the art.

[0125] Pharmaceutical compositions can be prepared by methods well known in the pharmaceutical arts. For example, therapeutic or pharmaceutical compositions intended to be administered by injection may contain one or more salts, buffers and / or stabilizers, along with sterile distilled water to form a solution. Surfactants may be added to promote the formation of a homogeneous solution or suspension. Surfactants are compounds that interact non-covalently with drugs to promote the dissolution or homogeneous suspension of the drug in an aqueous delivery system.

[0126] Therapeutic or pharmaceutical compositions may be administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the particular compound used; the metabolic stability and duration of action of the compound; the age, weight, general health, sex, and diet of the subject; the mode and time of administration; excretion rate; drug combination; the severity of the particular disorder or condition; and the subject being treated.

[0127] Also included are patient care kits comprising (a) an antibody that binds IL-18BP, as described herein; and, optionally, (b) at least one additional therapeutic agent. In certain kits, (a) and (b) are in separate therapeutic compositions. In some kits, (a) and (b) are in the same therapeutic composition.

[0128] The kits herein may also include one or more additional therapeutic agents or other components appropriate or desirable for the indication being treated or for the desired diagnostic application. The kits herein may also include one or more syringes or other components necessary or desirable to facilitate the intended mode of delivery (e.g., stent, implantable depot, etc.).

[0129] In some embodiments, the patient care kit contains separate containers, dividers, or compartments for the composition and informational material. For example, the composition may be contained in a bottle, vial, or syringe, and the informational material can be included in association with the container. In some embodiments, the separate elements of the kit are contained within a single, undivided container. For example, the composition is contained in a bottle, vial, or syringe having informational material in the form of a label attached thereto. In some embodiments, the kit contains multiple (e.g., packs) individual containers, each containing one or more unit dosage forms of the antibody (e.g., dosage forms described herein) and, optionally, at least one additional therapeutic agent. For example, the kit contains multiple syringes, ampoules, foil packets, or blister packs, each containing a single unit dose of the antibody and, optionally, at least one additional therapeutic agent. The containers of the kit can be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-tight.

[0130] The patient care kit optionally includes a device suitable for administering the composition, such as a syringe, inhaler, dropper (e.g., eye dropper), swab (e.g., cotton swab or wooden swab), or any such delivery device. In some embodiments, the device is an implantable device that dispenses a metered dose of the agent. Also included are methods of providing the kit, for example, by combining the components described herein.

[0131] Expression and purification system Certain embodiments include methods and related compositions for expressing and purifying the anti-IL-18BP antibodies described herein. Such recombinant anti-IL-18BP antibodies can be conveniently prepared using standard protocols described, for example, in Sambrook, et al. (1989, supra), especially sections 16 and 17; Ausubel et al. (1994, supra), especially chapters 10 and 16; and Coligan et al., Current Protocols in Protein Science (John Wiley & Sons, Inc. 1995-1997), especially chapters 1, 5, and 6. As one general example, an anti-IL-18BP antibody can be prepared by a procedure comprising one or more of the following steps: (a) preparing a construct comprising a polynucleotide sequence encoding an anti-IL-18BP antibody heavy and / or light chain and operably linked to regulatory elements; (b) introducing the construct into a host cell; (c) culturing the host cell to express the anti-IL-18BP antibody; and (d) isolating the anti-IL-18BP from the host cell.

[0132] Specific embodiments thus include polynucleotides encoding the anti-IL-18BP antibodies described herein, vectors containing the polynucleotides, and host cells containing the polynucleotides and / or vectors. To express the desired polypeptide, a nucleotide sequence encoding an anti-IL-18BP, or a functional equivalent, may be inserted into a suitable expression vector, i.e., a vector containing the necessary elements for the transcription and translation of the inserted coding sequence. Methods well known to those skilled in the art may be used to construct expression vectors containing a sequence encoding a polypeptide of interest and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Such techniques are described in Sambrook et al., Molecular Cloning, A Laboratory Manual (1989), and Ausubel et al., Current Protocols in Molecular Biology (1989).

[0133] A variety of expression vector / host systems are known and can be utilized to contain and express polynucleotide sequences, including, but not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus); plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems, including mammalian cells, more particularly human cell systems.

[0134] "Control elements" or "regulatory sequences" present in an expression vector are vector enhancers, promoters, and those untranslated regions of the 5' and 3' untranslated regions that interact with host cell proteins to effect transcription and translation. Such elements vary in their strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including constitutive and inducible promoters, can be used. For example, when cloning in bacterial systems, inducible promoters can be used, such as the hybrid lacZ promoter of the pBLUESCRIPT phagemid (Stratagene, La Jolla, CA) or the PSPORT1 plasmid (Gibco BRL, Gaithersburg, MD). In mammalian cell systems, promoters from mammalian genes or mammalian viruses are generally preferred. If it is necessary to generate cell lines containing multiple copies of a polypeptide-encoding sequence, SV40- or EBV-based vectors can be advantageously used with appropriate selectable markers.

[0135] In bacterial systems, numerous expression vectors can be selected depending on the intended use of the expressed polypeptide. For example, if large quantities are needed, vectors that direct high-level expression of easily purified fusion proteins can be used. Such vectors include, but are not limited to, multifunctional E. coli cloning and expression vectors, such as BLUESCRIPT (Stratagene), in which a sequence encoding a polypeptide of interest can be ligated into the vector in frame with the sequence for the amino-terminal Met and the following 7 residues of β-galactosidase, resulting in the production of a hybrid protein; pIN vectors (Van Heeke & Schuster, J. Biol. Chem. 264:5503-5509 (1989)); and the like. pGEX vectors (Promega, Madison, Wis.) can also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). Generally, such fusion proteins are soluble and can be easily purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins produced in such systems may be engineered to contain heparin, thrombin, or factor XA protease cleavage sites, allowing the cloned polypeptide of interest to be optionally released from the GST moiety.

[0136] Certain embodiments may use E. coli-based expression systems (see, e.g., Structural Genomics Consortium et al., Nature Methods. 5:135-146, 2008). These and related embodiments may rely partially or completely on ligation-independent cloning (LIC) to generate suitable expression vectors. In certain embodiments, protein expression may be controlled by T7 RNA polymerase (e.g., the pET vector series). These and related embodiments may utilize the expression host strain BL21(DE3), a λDE3 lysogen of BL21 that supports T7-mediated expression and is deficient in lon and ompT proteases for improved target protein stability. Also included are expression host strains that carry tRNA-encoding plasmids, which are rarely used in E. coli, such as the ROSETTA™(DE3) and Rosetta 2(DE3) strains. Cell lysis and sample handling can also be improved using reagents sold under the trademarks BENZONASE® nuclease and BUGBUSTER® protein extraction reagent. For cell culture, autoinduction media can improve the efficiency of many expression systems, including high-throughput expression systems. This type of media (e.g., the OVERNIGHT EXPRESS™ autoinduction system) gradually initiates protein expression through a metabolic shift without the addition of artificial inducers, such as IPTG. Certain embodiments use hexahistidine tags (such as those sold under the trademark HIS·TAG® fusions), followed by immobilized metal affinity chromatography (IMAC) purification or related techniques. However, in certain aspects, clinical-grade proteins can be isolated from E. coli inclusion bodies with or without the use of affinity tags (see, e.g., Shimp et al., Protein Expr Purif. 50:58-67, 2006). As a further example, certain embodiments may use a cold-shock inducible E. coli high-yield production system.This is because overexpression of proteins in E. coli at low temperatures improves their solubility and stability (see, e.g., Qing et al., Nature Biotechnology. 22:877-882, 2004).

[0137] Also included are high-density bacterial fermentation systems, such as high-cell-density cultures of Ralstonia eutropha, which allow protein production at cell densities exceeding 150 g / L and expression of recombinant proteins at titers exceeding 10 g / L.

[0138] In the yeast Saccharomyces cerevisiae, numerous vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PHH, can be used. For reviews, see Ausubel et al. (supra) and Grant et al., Methods Enzymol. 153:516-544 (1987). Also included is the Pichia pandoris expression system (see, e.g., Li et al., Nature Biotechnology. 24, 210-215, 2006; and Hamilton et al., Science, 301:1244, 2003). Certain embodiments include yeast systems engineered to selectively glycosylate proteins, including, inter alia, yeast with humanized N-glycosylation pathways (see, e.g., Hamilton et al., Science. 313:1441-1443, 2006; Wildt et al., Nature Reviews Microbiol. 3:119-28, 2005; and Gerngross et al., Nature-Biotechnology. 22:1409-1414, 2004; U.S. Patent Nos. 7,629,163; 7,326,681; and 7,029,872). By way of example only, recombinant yeast cultures can be grown in Fernbach flasks or 15 L, 50 L, 100 L, and 200 L fermentors, among others.

[0139] When a plant expression vector is used, expression of the polypeptide-encoding sequence can be driven by any of a number of promoters. For example, viral promoters, such as the 35S and 19S promoters of CaMV, can be used alone or in combination with the omega leader sequence from TMV (Takamatsu, EMBO J. 6:307-311 (1987)). Alternatively, plant promoters, such as the RUBISCO or small subunit heat shock promoters, can be used (Coruzzi et al., EMBO J. 3:1671-1680 (1984); Broglie et al., Science 224:838-843 (1984); and Winter et al., Results Probl. Cell Differ. 17:85-105 (1991)). These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. Such techniques are described in many publicly available reviews (see, eg, Hobbs in McGraw Hill, Yearbook of Science and Technology, pp. 191-196 (1992)).

[0140] Insect systems can also be used to express polypeptides of interest. For example, in one such system, Autographa californica nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes in Spodoptera frugiperda or Trichoplusia cells. A polypeptide-encoding sequence may be cloned into a non-essential region of the virus, such as the polyhedrin gene, and placed under control of the polyhedrin promoter. Successful insertion of the polypeptide-encoding sequence renders the polyhedrin gene inactive and produces recombinant virus lacking coat protein. The recombinant virus can then be used to infect, for example, S. frugiperda or Trichoplusia cells in which the polypeptide of interest can be expressed (Engelhard et al., Proc. Natl. Acad. Sci. USA 91:3224-3227 (1994)). Also included are baculovirus expression systems, including those utilizing SF9, SF21, and Tni cells (see, e.g., Murphy and Piwnica-Worms, Curr Protoc Protein Sci. Chapter 5:Unit 5.4, 2001). Insect systems can provide post-translational modifications similar to mammalian systems.

[0141] Many virus-based expression systems are commonly available for mammalian host cells.For example, when adenovirus is used as an expression vector, the sequence encoding the target polypeptide can be ligated into the adenovirus transcription / translation complex consisting of a late promoter and a tripartite leader sequence.Insertion into the non-essential E1 or E3 region of the viral genome can also be used to obtain a viable virus capable of expressing polypeptides in infected host cells (Logan & Shenk, Proc. Natl. Acad. Sci. USA 81:3655-3659 (1984)).Transcription enhancers, such as Rous sarcoma virus (RSV) enhancers, can also be used to increase expression in mammalian host cells.

[0142] Examples of useful mammalian host cell lines are the SV40-transformed monkey kidney CV1 line (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); mouse Sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat hepatocytes (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TR1 cells (Mather et al., Annals NYAcad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and human hepatoma line (Hep G2). Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al., PNAS USA 77:4216 (1980)); and myeloma cell lines, such as NSO and Sp2 / 0. For a review of suitable specific mammalian host cell lines for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 255-268. Certain preferred mammalian cell expression systems include CHO and HEK293-cell based expression systems.Mammalian expression systems can utilize adherent cell lines in, for example, T-flasks, roller bottles, or cell factories, or suspension cultures, such as 1 L and 5 L spinners, 5 L, 14 L, 40 L, 100 L, and 200 L stirred tank bioreactors, or 20 / 50 L and 100 / 200 L WAVE bioreactors, among others known in the art.

[0143] Also included is cell-free expression of proteins. These and related embodiments typically utilize purified RNA polymerase, ribosomes, tRNA, and ribonucleotides; these reagents may be produced by extraction from cells or from cell-based expression systems.

[0144] Specific initiation signals can also be used to achieve more efficient translation of sequences encoding a polypeptide of interest. Such signals include the ATG initiation codon and adjacent sequences. When a polypeptide-encoding sequence, its initiation codon, and upstream sequences are inserted into an appropriate expression vector, additional transcriptional or translational control signals may not be required. However, when only a coding sequence, or a portion thereof, is inserted, exogenous translational control signals, including the ATG initiation codon, should be provided. Furthermore, the initiation codon should be in the correct reading frame to ensure translation of the entire insert. Exogenous translational elements and initiation codons can be of various origins, both natural and synthetic. Expression efficiency can be enhanced by the inclusion of enhancers appropriate for the particular cell system used, such as those described in the literature (Scharf et al., Results Probl. Cell Differ. 20:125-162 (1994)).

[0145] A host cell line may also be selected for its ability to regulate the expression of inserted sequences or process the expressed protein in the desired manner. Such modifications of polypeptides include, but are not limited to, post-translational modifications, such as acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing, which cleaves a "prepro" form of the protein, may be used to facilitate correct insertion, folding, and / or function. Different host cells, such as yeast, CHO, HeLa, MDCK, HEK293, and W138, as well as bacterial cells that possess or lack specific cellular and characteristic machinery for such post-translational activities, may be selected to ensure the correct modification and processing of the foreign protein.

[0146] For long-term, high-yield production of recombinant proteins, stable expression is generally preferred. For example, cell lines stably expressing a polynucleotide of interest can be transformed using an expression vector, which may contain a viral origin of replication and / or endogenous expression elements, as well as a selectable marker gene on the same or a separate vector. After introduction of the vector, the cells may be grown in an enriched medium for approximately 1-2 days before switching them to a selective medium. The purpose of the selectable marker is to confer resistance to selection; its presence allows the growth and recovery of cells that successfully express the introduced sequence. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate for the cell type. Transient production, such as by transient transfection or infection, can also be used. Exemplary mammalian expression systems suitable for transient production include HEK293 and CHO-based systems.

[0147] Any number of selection systems may be used to recover transformed or transduced cell lines, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al., Cell 11:223-232 (1977)) and adenine phosphoribosyltransferase (Lowy et al., Cell 22:817-823 (1990)) genes, which can be used in tk- or aprt- cells, respectively. Antimetabolite, antibiotic, or herbicide resistance can also be used as the basis for selection; for example, dhfr, which confers resistance to methotrexate (Wigler et al., PNAS USA. 77:3567-70 (1980)); npt, which confers resistance to aminoglycosides, neomycin, and G-418 (Colbere-Garapin et al., J. Mol. Biol. 150:1-14 (1981)); and als or pat, which confers resistance to chlorsulfuron and phosphinothricin acetyltransferase, respectively (Murry, supra). Additional selectable genes have been described, such as trpB, which allows cells to utilize indole instead of tryptophan, or hisD, which allows cells to utilize histinol instead of histidine (Hartman & Mulligan, Proc. Natl. Acad. Sci. USA 85:8047-51 (1988)). The use of visible markers has gained popularity, with markers such as green fluorescent protein (GFP) and other fluorescent proteins (e.g., RFP, YFP), anthocyanins, β-glucuronidase and its substrate GAS, and luciferase and its substrate luciferin being widely used not only to identify transformants but also to quantify the amount of transient or stable protein expression resulting from a particular vector system (see, e.g., Rhodes et al., Methods Mol. Biol. 55:121-131 (1995)).

[0148] Also included are high-throughput protein production systems, or microproduction systems. Certain embodiments may utilize hexahistidine fusion tags for protein expression and purification on metal chelate-modified slide surfaces or MagneHis Ni-particles (see, e.g., Kwon et al., BMC Biotechnol. 9:72, 2009; and Lin et al., Methods Mol Biol. 498:129-41, 2009). Also included are high-throughput cell-free protein expression systems (see, e.g., S Sitaraman et al., Methods Mol Biol. 498:229-44, 2009). These and related embodiments can be used, for example, to generate antibody microarrays, which can then be used to screen libraries to identify antibodies and antigen-binding domains that interact with the IL-18BP polypeptide of interest.

[0149] Although the foregoing embodiments have been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in view of the teachings of the present disclosure that certain changes and modifications can be made without departing from the spirit or scope of the appended claims. The following examples are provided by way of illustration only, and not by way of limitation. Those of ordinary skill in the art will readily recognize a variety of non-critical parameters that can be changed or modified to yield essentially similar results.

[0150] Listing of Exemplary Embodiments The following non-limiting enumerated embodiments are provided by way of example.

[0151] Embodiment I-1. An antibody specific to interleukin-18 binding protein (IL-18BP), which interferes with the binding of IL-18 to IL-18BP.

[0152] Embodiment I-2. The antibody of embodiment I-1, wherein the antibody binds to a conformational epitope of IL-18BP.

[0153] Embodiment I-3. The antibody of embodiment I-2, wherein the antibody binds to two or more of amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372.

[0154] Embodiment I-4. The antibody of embodiment I-2, wherein the antibody binds to amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372.

[0155] Embodiment I-5. The antibody of embodiment I-1, wherein the antibody binds to a linear epitope of IL-18BP.

[0156] Embodiment I-6. The antibody of any one of embodiments I-1 to I-5, wherein the antibody binds to the binding interface between IL-18 and mature IL-18BP.

[0157] Embodiment I-7. The antibody of embodiment I-6, wherein the antibody binds to amino acid residues S75, H79, T116, S119 of SEQ ID NO: 372.

[0158] Embodiment I-8. The antibody of any one of embodiments 1 to 7, wherein the antibody binds to human IL-18BP and cynomolgus IL-18BP, but does not bind to mouse IL-18BP.

[0159] Embodiment I-9. The antibody of any one of embodiments I-1 to I-7, wherein the antibody binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

[0160] Embodiment I-10. The antibody was designed to measure the binding affinity (K D The antibody of any one of embodiments I-1 to I-9, which binds to IL-18BP with a binding affinity of greater than about 650 pM, optionally with a binding affinity of between about 1 pm and about 650 pm, or about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM or less.

[0161] Embodiment I-11. The antibody of any one of embodiments I-1 to I-10, wherein the antibody is an IL-18BP antagonist, antagonizing the binding activity between IL-18BP and IL-18.

[0162] Embodiment I-12. The antibody of embodiment I-11, wherein the antibody blocks the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signaling, including induction of IFN-gamma, CXCL10 and / or TNFα.

[0163] Embodiment I-13. The antibody of any one of embodiments I-1 to I-12, comprising an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or hybrids and / or variants thereof.

[0164] Embodiment I-14. The antibody of embodiment I-13, comprising an IgG Fc domain with enhanced effector function in humans, optionally an IgG1 or IgG3 Fc domain.

[0165] Embodiment I-15. The antibody of embodiment I-13, which comprises an IgG Fc domain with reduced effector function in humans, optionally an IgG2 or IgG4 Fc domain.

[0166] Embodiment I-16. The antibody of any one of embodiments I-1 to I-15, wherein the antibody is a monoclonal antibody.

[0167] Embodiment I-17. The antibody of any one of embodiments I-1 to I-16, wherein the antibody is a humanized antibody.

[0168] Embodiment I-18. The antibody of any one of embodiments I-1 to I-17, wherein the antibody is an scFv.

[0169] Embodiment I-19. The antibody is a) a V complementarity-determining region selected from Table A1 H CDR1 sequence, V H CDR2 sequence, and V H The heavy chain variable region (V H ), and variants thereof that specifically bind to IL-18BP; and b) a V complementarity-determining region selected from Table A1 L CDR1 sequence, V L CDR2 sequence, and V L The light chain variable region (V L ), as well as variants thereof that specifically bind to IL-18BP.

[0170] Embodiment I-20. a)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V Lthe CDR3 sequences comprise SEQ ID NOs: 4 to 6, respectively; b)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 7 to 9, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 10 to 12, respectively; c)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 13 to 15, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 16 to 18, respectively; d)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 19 to 21, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 22 to 24, respectively; e)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 25 to 27, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 28 to 30, respectively; f)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 31 to 33, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 34 to 36, respectively; g)V H CDR1 sequence, VH CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 37 to 39, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 40 to 42, respectively; h)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 43 to 45, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 46 to 48, respectively; i)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 49 to 51, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 52 to 54, respectively; j)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 55 to 57, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 58 to 60, respectively; k)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 61 to 63, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 64 to 66, respectively; l)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 67 to 69, respectively, and VL CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 70 to 72, respectively; m)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 73 to 75, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 76 to 78, respectively; n)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 79 to 81, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 82 to 84, respectively; o)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 85 to 87, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 88 to 90, respectively; p)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 91 to 93, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 94 to 96, respectively; q)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 97 to 99, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V Lthe CDR3 sequences comprise SEQ ID NOs: 100 to 102, respectively; r)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 103 to 105, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 106-108, respectively; s)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 109 to 111, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; t)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 115 to 117, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; u)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 121 to 123, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 124 to 126, respectively; v)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 127 to 129, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 130 to 132, respectively; w)VH CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 133 to 135, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 136 to 138, respectively; x)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 139 to 141, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 142 to 144, respectively; y)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 145 to 147, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 148 to 150, respectively; z)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 151 to 153, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 154 to 156, respectively; aa)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 157 to 159, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 160 to 162, respectively; bb)V H CDR1 sequence, V H CDR2 sequence, and VH The CDR3 sequences include SEQ ID NOs: 163 to 165, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; cc)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 169 to 171, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 172 to 174, respectively; dd)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 175 to 177, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 178 to 180, respectively; ee)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 181 to 183, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 184 to 186, respectively; ff)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 187 to 189, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 190 to 192, respectively; gg)V H CDR1 sequence, V H CDR2 sequence, and V HThe CDR3 sequences include SEQ ID NOs: 193 to 195, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; hh)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 199 to 201, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; ii) V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 205 to 207, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 208-210, respectively; jj)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 211 to 213, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 214-216, respectively; kk)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 217 to 219, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; ll)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 223 to 225, respectively, and VL CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; mm)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 229 to 231, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 232 to 234, respectively; nn)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 235 to 237, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; oo)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 241 to 243, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 244 to 246, respectively; pp)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 247 to 249, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 250 to 252, respectively; qq)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences include SEQ ID NOs: 253 to 255, respectively, and V L CDR1 sequence, V LCDR2 sequence, and V L the CDR3 sequences comprise SEQ ID NOs: 256-258, respectively; or rr)V H CDR1 sequence, V H CDR2 sequence, and V H The CDR3 sequences comprise SEQ ID NOs: 259 to 261, respectively, and V L CDR1 sequence, V L CDR2 sequence, and V L The antibody of embodiment I-19, wherein the CDR3 sequences comprise SEQ ID NOs: 262-264, respectively.

[0171] Embodiment I-21. V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V H 21. The antibody of any one of embodiments 1 to 20, having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

[0172] Embodiment I-22. V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally, V L The antibody of any one of embodiments I-1 to I-21, having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

[0173] Embodiment I-23. a)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 265, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 266; b)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 267, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 268; c)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 269, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 270; d)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 271, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 272; e)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 273, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 274; f)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 275, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 276; g)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; h)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; i)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 281, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 282; j)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 283, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 284; k)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 285, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 286; l)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 287, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 288; m)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 289, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 290; n)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 291, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 292; o)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 293, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 294; p)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 295, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 296; q)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 297, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 298; r)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 299, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 300; s)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 301, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 302; t)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 303, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 304; u)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 305, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 306; v)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 307, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 308; w)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 309, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 310; x)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 311, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 312; y)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 313, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 314; z)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 315, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 316; aa)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 317, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 318; bb)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 319, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 320; cc)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 321, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 322; dd)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 323, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 324; ee)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 325, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 326; ff)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 327, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 328; gg)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 329, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 330; hh)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 331, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 332; ii) V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334; jj)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 335, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 336; kk)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 337, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 338; ll)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 339, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 340; mm)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 341, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 342; nn)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 343, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 344; oo)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 345, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 346; pp)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 347, and V L comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 348; qq)V H comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 349, and V L comprises a sequence that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 350; or rr)V Hcomprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 351, and V L The antibody of any one of embodiments I-1 to I-22, comprising a sequence that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 352.

[0174] Embodiment I-24. An isolated polynucleotide encoding the isolated anti-IL-18BP antibody according to any one of embodiments I-1 to I-23, an expression vector comprising the isolated polynucleotide, or an isolated host cell comprising the vector.

[0175] Embodiment I-25. A pharmaceutical composition comprising an anti-IL-18BP antibody according to any one of embodiments I-1 to I-23, and a pharmaceutically acceptable carrier.

[0176] Embodiment I-26. The pharmaceutical composition of embodiment I-25, wherein the composition is optionally a sterile injectable solution suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

[0177] Embodiment I-27. A method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of embodiments I-25 to I-26.

[0178] Embodiment I-28. The method of embodiment I-27, wherein the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, optionally a proliferative disease or disorder selected from a lymphoproliferative disorder, a myeloproliferative disorder, a proliferative enterocolitis, a proliferative diabetic retinopathy, and a proliferative kidney disease.

[0179] Embodiment I-29. The method of embodiment I-28, wherein the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18.

[0180] Embodiment I-30. Cancers include bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (liver cell carcinoma), sarcoma, B-cell malignancies, breast cancer, The method of embodiment I-28 or I-29, wherein the cancer is selected from one or more of: ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, urothelial cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer.

[0181] Embodiment I-31. The method according to any one of embodiments I-27 to I-30, comprising administering the pharmaceutical composition according to embodiment I-25 or I-26 in combination with IL-18.

[0182] Embodiment I-32. The method of any one of embodiments I-28 to I-31, comprising administering the pharmaceutical composition of embodiment I-25 or I-26 in combination with an immune checkpoint modulator selected from an antagonist of an inhibitory immune checkpoint molecule, and an agonist of a stimulatory immune checkpoint molecule.

[0183] Embodiment I-33. The method of embodiment I-32, wherein the immune checkpoint modulator is a polypeptide, optionally an antibody, or a ligand, or a small molecule.

[0184] Embodiment I-34. The method of embodiment I-32 or I-33, wherein the inhibitory immune checkpoint molecule is selected from one or more of Programmed Death-Ligand 1 (PD-L1), Programmed Death 1 (PD-1), Programmed Death-Ligand 2 (PD-L2), Cytotoxic T-Lymphocyte-Associated protein 4 (CTLA-4), Indoleamine 2,3-dioxygenase (IDO), Tryptophan 2,3-dioxygenase (TDO), T-cell Immunoglobulin domain and Mucin domain 3 (TIM-3), Lymphocyte Activation Gene-3 (LAG-3), V-domain Ig suppressor of T cell activation (VISTA), B and T Lymphocyte Attenuator (BTLA), CD160, Herpes Virus Entry Mediator (HVEM), and T-cell immunoreceptor with Ig and ITIM domains (TIGIT).

[0185] Embodiment I-35. a) the antagonist is an antibody or small molecule that specifically binds to PD-L1 and / or PD-L2, an antagonist of PD-L1 and / or PD-L2 optionally chosen from one or more of atezolizumab (MPDL3280A), avelumab (MSB0010718C) and durvalumab (MEDI4736), optionally in which the cancer is selected from one or more of colorectal cancer, melanoma, breast cancer, non-small cell lung cancer, bladder cancer and renal cell carcinoma; b) the antagonist is a PD-1 antagonist optionally chosen from one or more of an antibody or small molecule that specifically binds to PD-1, nivolumab, pembrolizumab, MK-3475, AMP-224, AMP-514PDR001, and pidilizumab, optionally in which the PD-1 antagonist is nivolumab, and the cancer is optionally chosen from one or more of Hodgkin lymphoma, melanoma, non-small cell lung cancer, hepatocellular carcinoma, renal cell carcinoma, and ovarian cancer; c) the PD-1 antagonist is pembrolizumab, and the cancer is optionally selected from one or more of melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, and urothelial cancer; d) the antagonist is a CTLA-4 antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to CTLA-4, ipilimumab, tremelimumab, optionally in which the cancer is selected from melanoma, prostate cancer, lung cancer, and bladder cancer; e) the antagonist is an IDO antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to IDO, indoximod (NLG-8189), 1-methyl-l-tryptophan (1MT), β-carboline (norharman; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat, wherein the cancer is optionally selected from one or more of metastatic breast cancer and brain cancer, optionally glioblastoma multiforme, glioma, gliosarcoma, or malignant brain tumor; f) the antagonist is a TDO antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to TDO, 680C91, and LM10; g) the antagonist is a TIM-3 antagonist optionally selected from one or more of an antibody or a small molecule that specifically binds to TIM-3; h) the antagonist is a LAG-3 antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to LAG-3, and BMS-986016; i) the antagonist is a VISTA antagonist optionally selected from one or more of an antibody or a small molecule that specifically binds to VISTA; j) the antagonist is an antagonist of BTLA, CD160 and / or HVEM, optionally selected from one or more of an antibody or a small molecule that specifically binds to BTLA, CD160 and / or HVEM; or k) The method of embodiment I-34, wherein the antagonist is a TIGIT antagonist optionally selected from one or more of an antibody or a small molecule that specifically binds to TIGIT.

[0186] Embodiment I-36. The method of embodiment I-32 or I-33, wherein the stimulatory immune checkpoint molecule is selected from one or more of OX40, CD40, Glucocorticoid-Induced TNFR Family Related Gene (GITR), CD137 (4-1BB), CD27, CD28, CD226, and Herpes Virus Entry Mediator (HVEM).

[0187] Embodiment I-37. a) the agonist is an OX40 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to OX40, OX86, Fc-OX40L, and GSK3174998; b) the agonist is a CD40 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD40, CP-870,893, dacetuzumab, Chi Lob 7 / 4, ADC-1013, and rhCD40L, wherein the cancer is optionally selected from one or more of melanoma, pancreatic cancer, mesothelioma, and hematological cancers, optionally lymphomas such as non-Hodgkin's lymphoma; c) the agonist is a GITR agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to GITR, INCAGN01876, DTA-1, and MEDI1873; d) the agonist is a CD137 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD137, utomilumab, and 4-1BB ligand; e) the agonist is a CD27 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD27, varlilumab, and CDX-1127 (1F5); f) the agonist is a CD28 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD28, and TAB08; and / or g) The method of embodiment I-36, wherein the agonist is an HVEM agonist optionally selected from one or more of an antibody or a small molecule or a ligand that specifically binds to HVEM.

[0188] Embodiment I-38. The method according to any one of embodiments I-28 to I-37, comprising administering the pharmaceutical composition according to embodiment I-25 or I-26 in combination with at least one chemotherapeutic agent.

[0189] Embodiment I-39. The method of embodiment I-38, wherein the at least one chemotherapeutic agent is selected from one or more of an alkylating agent, an antimetabolite, a cytotoxic antibiotic, a topoisomerase inhibitor (type 1 or type II), and an anti-microtubule agent.

[0190] Embodiment I-40. a) the alkylating agent is selected from one or more of nitrogen mustards (optionally mechlorethamine, cyclophosphamide, mustine, melphalan, chlorambucil, ifosfamide, and busulfan), nitrosoureas (optionally N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, and streptozotocin), tetrazines (optionally dacarbazine, mitozolomide, and temozolomide), aziridines (optionally thiotepa, mitomycin, and diaziquone (AZQ)), cisplatin and its derivatives (optionally carboplatin and oxaplatin), and non-classical alkylating agents (optionally procarbazine and hexamethylmelamine); b) the antimetabolite is selected from one or more of antifolates (optionally methotrexate and pemetrexed), fluoropyrimidines (optionally 5-fluorouracil and capecitabine), deoxynucleoside analogs (optionally ancitabine, enocitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, fludarabine, and pentostatin), and thiopurines (optionally thioguanine and mercaptopurine); c) the cytotoxic antibiotic is selected from one or more of anthracyclines (optionally doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, and mitoxantrone), bleomycin, mitomycin C, mitoxantrone, and actinomycin; d) the topoisomerase inhibitor is selected from one or more of camptothecin, irinotecan, topotecan, etoposide, doxorubicin, mitoxantrone, teniposide, novobiocin, mervalone, and aclarubicin; and / or e) The method of embodiment I-39, wherein the anti-microtubule agent is selected from one or more of taxanes (optionally paclitaxel and docetaxel) and vinca alkaloids (optionally vinblastine, vincristine, vindesine, vinorelbine).

[0191] Embodiment I-41. The method of embodiment I-27, wherein the disease or condition is myelodysplastic syndrome (MDS).

[0192] Embodiment I-42. The method of embodiment I-27, wherein the disease or condition is an infectious disease.

[0193] Embodiment I-43. The method of embodiment I-42, wherein the infectious disease is selected from viral, bacterial, fungal (optionally yeast), and protozoal infections.

[0194] Embodiment I-44. The method according to any one of embodiments I-41 to I-43, comprising administering the pharmaceutical composition according to embodiment I-25 or I-26 in combination with IL-18.

[0195] Embodiment I-45. 1. A method of screening anti-IL-18BP antibodies for their ability to block or inhibit binding between IL-18 and IL-18BP, comprising: a) determining the binding affinity of an antibody to (i) IL-18BP alone, and (ii) an IL-18-reduced fusion protein, the IL-18-reduced fusion protein comprising IL-18 fused to IL-18BP via a flexible linker (and an optional protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein is bound to the IL-18BP portion of the fusion protein and sterically blocks the IL-18 binding site of the IL-18BP portion of the fusion protein; b) comparing the binding affinity of (i) with the binding affinity of (ii); and c) identifying or selecting an antibody as being capable of blocking or inhibiting the binding between IL-18 and IL-18BP if the binding affinity of (i) is significantly stronger than the binding affinity of (ii).

[0196] Embodiment I-46. The method of embodiment I-45, wherein the IL-18 and IL-18BP are murine IL-18 and IL-18BP.

[0197] Embodiment I-47. The method of embodiment I-45, wherein the IL-18 and IL-18BP are human IL-18 and IL-18BP. [Example]

[0198] Example 1: Generation of antagonistic monoclonal antibodies against interleukin-18 binding protein (IL-18BP) We conducted studies to generate potent human and humanized antibodies that bind to and inhibit the IL-18 / IL-18BP complex, thereby liberating IL-18 and stimulating immune activity. Potential therapeutic candidates with cross-reactivity to human and cynomolgus IL-18BP were identified. Furthermore, we generated potent anti-mouse IL-18BP mAbs closely related to their anti-human / cynomolgus counterparts, enabling us to investigate these agents in mouse tumor model systems.

[0199] material and method Immunization and isolation of antigen-specific antibodies from single B cells. Monoclonal antibodies against IL-18BP were generated by immunizing mice with IL-18BP, followed by isolation of antigen-specific single B cells using a Berkeley Lights Beacon instrument and cloning the antibody-encoding gene from each cell. Mouse mAbs were generated using autoimmune mouse strains, DiversimAb™ and DivergimAb™ (Abveris Inc.). Mice were immunized with human IL-18BP and cynomolgus IL-18BP (tagged with either his or Fc) on an alternating schedule, while titers were tested with human, cynomolgus, or mouse IL-18BP using the other tag in each case to avoid detection of tag-specific antibodies.

[0200] After high titering against IL-18BP, antigen-specific B cells from appropriate mice were isolated as single cells using a Berkeley Lights Beacon instrument, using the manufacturer's recommended procedures as described in the literature (Mullen et al., Antibody Therapeutics. 4(3):185-196, 2021). The immunoglobulin gene sequences of the antigen-specific B cells were obtained and used to generate recombinant antibodies using established methods.

[0201] Antibody naming. Some antibodies are named using the prefix "SA," a two-digit sequence, and a single-letter suffix. The suffix indicates the identity of the constant domain of the HC, such as "a" for human IgG1 and "d" for mouse IgG2a. Thus, SA04a is a human IgG1 antibody.

[0202] Variants and mutations were named in the following order: original amino acid, followed by Kabat position number (Kabat 1991), followed by the substituted amino acid. Standard single-letter codes for amino acids were used. Thus, Y32E indicates that tyrosine (Y) at Kabat position 32 has been replaced by glutamic acid (E).

[0203] Library / site-directed mutagenesis preparation. To affinity-maturate the starting mAb, a library of variants was prepared, focusing on the HC CDR and then each of the LC CDRs. To do this, amino acids in each CDR were then replaced with up to 17 amino acid substitutions. To avoid introducing potential unwanted sequence defects, cysteines and tryptophans were not included in the library, and parent amino acids already in place were not included in the screening. To generate variants at each position, two sets of mutagenic oligonucleotides (Integrated DNA Technologies (IDT), San Diego) containing the degenerate codons NDT or VHG (where N = A / C / G / T, D = A / G / T, V = A / C / G, and H = A / C / T) were used for each position (Kille, 2013; Acevedo-Rocha, 2015) and paired with appropriate 5' and 3' distal oligonucleotides (IDT) designed to allow amplification and cloning.

[0204] PCR was performed using high-fidelity DNA polymerase (Q5, New England Biolabs) according to the manufacturer's protocol. The parental plasmids (both heavy and light chains) were diluted to 10 ng / μL, and 1 μL of each was used as template in a 50 μL reaction. Gene fragments for the V regions with the degenerate codons described above were amplified by PCR and purified (Qiagen PCR purification kits were used according to the manufacturer's instructions). The gene fragments were assembled into heavy and light chain clones by either overlap extension PCR (OE-PCR) or Gibson cloning. For OE-PCR, fragments were amplified using corresponding forward and reverse primers containing restriction enzyme sites (AgeI-NheI for the heavy chain, SbfI-MfeI for the light chain) and column-purified (Qiagen PCR purification kits). Restriction enzyme digestion was performed using high-fidelity enzymes (New England Biolabs), and the fragments were ligated into the appropriate heavy and light chain vectors using T4 DNA ligase (New England Biolabs, catalog number M0202L). The empty heavy chain vector contains most of the human IgG1 constant region with an engineered NheI site (created by changing the wobble position) 12 amino acids within the constant region. The empty light chain vector contains most of the human kappa constant region with an engineered MfeI site 18 amino acids within the constant region. Gibson cloning was performed using the same empty vector fragments (Gibson Assembly® Master Mix Kit, New England Biolabs, catalog number ES2611L). Inserts were normalized to 1 ng / μL, and a total of 2 ng of insert DNA was used (1 ng per fragment). The 10 μL reaction volume consisted of 5 μL of Gibson Master Mix and QS with purified water. The reaction was incubated at 50°C for 15-60 minutes.

[0205] Ligation products from either OE-PCR or Gibson assembly were transformed into competent E. coli (Monserate Biotechnology, San Diego) by adding 2–5 μL of the ligation reaction mixture to the cells and incubating on ice for 5 minutes. The cells were heat shocked at 42°C for 30 seconds and placed on ice. 250 μL of SOC medium (BioPioneer, San Diego or Teknova, San Diego) was added, and the tube was incubated at 37°C and 200 rpm for 1 hour. 100 μL of the culture was plated onto antibiotic selection plates (BioPioneer, San Diego or Teknova, San Diego) and incubated overnight at 37°C.

[0206] Plates were sent for antibody gene colony sequencing (Genewiz or Eton). 24–48 clones per plate were sequenced. Sequences were analyzed against a reference sequence using SnapGene Software (GSL Biotech). This was an in silico cloning of the library. Clones were selected based on sequence alignment and the amino acids encoded by the mutation primers, and used to perform individual plasmid minipreps.

[0207] Mutant screening was performed by small-scale expression of library-derived plasmids in Expi293F cells cultured in 48-well plates using the method described below. Heavy and light chain pairing was performed at a 1:2 ratio (0.5 ng HC:1 ng LC plasmid per well). Replicates of the parental antibody control were included on each plate. Cells were grown in the plates for 3 days, after which the supernatant from each well was harvested and screened.

[0208] Screening of plate transfections was performed using biolayer interferometry (BLI). First, the concentration of antibody present in each sample was determined using BLI, and then the antibodies were screened for binding affinity to IL-18BP. Initial apparent binding kinetics measurements were performed on a ForteBio Octet RED96e instrument. mAbs were loaded onto an anti-human constant domain (AHC) biosensor (ForteBio) in 10x kinetics buffer consisting of PBS, 0.1% BSA, and 0.02% Tween 20 for 120 seconds, yielding spectral shift values ​​of 0.8–1.2 nm. The association phase was performed in the presence of 20 nM human, cynomolgus monkey, or mouse IL-18BP ortholog and allowed to proceed for 120 seconds. Dissociation was measured for 300 seconds to determine whether the variants exhibited improved binding or dissociation rates compared to the parent antibody. Candidates that showed apparent improved binding kinetics based on one screening concentration were then retested for overall binding kinetics, compared to their respective orthologs, and recombined with other mutations as described below.

[0209] Recombinant antibody expression and purification. Expi293F cells from the Expi293 Expression System Kit (Thermo Fisher Scientific, Cat. No. A14635) were grown in Expi293F Expression Medium (Cat. No. A1435101). 3–6 × 10 6 Cells were grown to a density of 1000 cells / mL and then counted using a hemocytometer. Plasmid DNA (1.0 μg per 1.0 mL of culture) was diluted in Opti-MEM Reduced Serum Medium (RSM) (Cat. No. 31985062). The Opti-MEM RSM value was obtained from the manufacturer's transfection recommendations. Expifectamine 293 reagent was diluted in Opti-MEM RSM, incubated for 5 minutes at room temperature, and then mixed with the diluted plasmid DNA. This mixture was allowed to incubate at room temperature for 10-20 minutes. While the expifectamine / plasmid DNA complex was incubating, Expi293F cells were transfected at 3 x 10 6The cells were diluted to a density of 1000 cells / mL and added to an Erlenmeyer flask of the desired volume (BioPioneer, 125 mL flasks: DGFPC0125S). The expifectamine / plasmid DNA complex was then slowly transferred to the shake flask containing the Expi293F cells, and the flask was placed in a shaking incubator with a 25 mm orbital throw (Infors-HT Multitron) at 37°C, 8% CO2, and 125 rpm. 18–22 hours after transfection, ExpiFectamine 293 Transfection Enhancer 1 (#100013863) and 2 (#A14350-01) were added to the cells, and the cells were returned to the shaking incubator. The cells were then incubated for an additional 4 days, after which they were purified by centrifugation at 4000 × g for 20 minutes in a refrigerated centrifuge and 0.22 μm filtration.

[0210] Antibodies were purified using a 5 mL HiTrap MabSelect SuRe (Protein A) column on an AKTA Explorer FPLC system. Residual bound protein was first removed from the column by adding 50 mL of 0.1 M glycine, pH 3.0 (elution buffer), followed by 50 mL of 50 mM glycine, 50 mM glycinate, pH 8 (binding / wash buffer). The antibody (25–400 mL) was loaded onto the column at 5 mL / min and further washed with 25 mL of equilibration / wash buffer until the UV reading reached baseline. The mAb was then eluted using a 25 mL linear gradient of 0–100% elution buffer over 2 min at 5 mL / min. Antibody elution was monitored by absorbance at 280 nm. Peak fractions were collected and pooled into a 15 mL conical tube. The material was then buffer exchanged into storage buffer (PBS, pH 7.4) using a PD10 column (Cytiva catalog number 17085101). It was then filter-sterilized (GenClone Syringe Filters, Cat. No. 25-244, attached to BD 5 mL [Cat. No. 309646] and 20 mL [Cat. No. 302830] BD Luer-Lok™ syringes) into 15 mL conical tubes for subsequent characterization.

[0211] Analysis by size exclusion HPLC (SEC-HPLC). SEC-HPLC was performed using a 5 μm particle size, 7.8 mm ID x 30 cm TSKgel G3000SW. XL The SEC-HPLC analysis was performed at 280 nm using 50 mM sodium phosphate, 200 mM arginine, pH 6.8, isocratically on an Agilent 1100 HPLC at a flow rate of 1 mL / min. Detection was performed at 280 nm using a diode array detector, and peaks were integrated using Agilent ChemStation software. SEC-HPLC standards used to calibrate the column consisted of bovine thyroglobulin, bovine IgG, chicken albumin, bovine ribonuclease A, and p-aminobenzoic acid (Sigma-Aldrich, catalog number 69385).

[0212] Analysis of binding affinity by biolayer interferometry (BLI). Binding kinetics measurements were performed on a Fortebio (now Sartorius) Octet RED96e instrument. mAbs were loaded onto anti-human constant domain (AHC) biosensors (ForteBio) for 90–120 s in 10x kinetics buffer consisting of PBS containing 0.1% BSA and 0.02% Tween 20, yielding spectral shift values ​​of 0.8–1.2 nm. Binding was performed in the presence of a two-fold dilution series of hIL-18BP and typically proceeded over 90–120 s. Dissociation was generally measured over 300–1200 s. The dilution series started at 100 nM for the weaker variants and 10 nM for the most potent mAb. Cross-reactivity to cynomolgus monkey IL-18BP and mouse IL-18BP was determined using the same method as for the IL-18BP of the appropriate species.

[0213] Activity of anti-IL-18BP mAb in IL-18 reporter HEK293 cells. InvivoGen (hkb-hmil18) IL-18 reporter HEK293 cells respond to exogenously added IL-18 by expressing the NF-κB / AP-1-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene. To perform this assay, cells were grown in complete DMEM medium (10% HI FBS, 1% PS) containing 1x HEK-Blue Selection (Invivogen, hb-sel). Cells were carefully rinsed twice with 1x PBS and lifted in 1x PBS at 37°C for 5 minutes. Cells were counted, spun down at 200 x g for 5 minutes, and 2.5 x 10 5The cells were resuspended in non-selective complete DMEM at 25,000 cells / mL. The cells were then seeded at 25,000 cells / mL in 100 μL / well of a 96-well plate (Genesee Scientific, 25-109). The plated cells were then placed in a 37°C, 5% CO2 incubator. Test antibody solutions were prepared at 90 μg / mL in non-selective DMEM and incubated with 120 ng / mL human IL-18BP at room temperature for 30 minutes. A 0.6 ng / mL stock of recombinant human IL-18 (Sino Biological, 10119-HNCE) was then prepared and added to the antibody / IL-18BP complex after incubation. The resulting solution was immediately added to the cells to achieve final concentrations of 15 μg / mL test antibody, 20 ng / mL human IL-18BP, and 0.1 ng / mL human IL-18. Cells were incubated at 37°C, 5% CO for 18-22 hours. 10 μL / well of cell supernatant was collected and mixed with 90 μL / well of complete Quanti-Blue solution (Invivogen) in a new 96-well plate and placed in an incubator at 37°C for 1-3 hours, noting the colorimetric change from purple to blue. Plates were then read at 620 nm, and data were analyzed using GraphPad Prism.

[0214] Inhibition of IFNγ expression by anti-IL-18BP mAb in KG-1 cells. Human KG-1 cells (ATCC catalog no. CCL-246) were seeded at 150k cells / well. IL-18BP (final concentration 50ng / mL, Sino Biologicals, catalog no. 10357-H08H) was pre-blocked with serial dilutions of antibody for 20 minutes at room temperature. IL-18 (final concentration 10ng / mL, R&D Systems, catalog no. 9124-IL / CF) was added to the mixture and incubated for an additional 20 minutes at room temperature. The mixture was added to the cells and incubated overnight at 37°C. Secreted IFNγ in the cell culture supernatant was measured using the R&D Systems Human IFN-gamma DuoSet (catalog no. DY285B) in a MaxiSorp flat-bottom plate (catalog no. 44-2404-21). The supernatant was diluted 1:2 with reagent diluent according to the manufacturer's protocol. Absorbance was measured at 450 nm using a Spectramax iD5 plate reader. Data were analyzed using GraphPad Prism.

[0215] PBMC assay for human IL-18 activity. Human peripheral blood mononuclear cells (PBMCs) obtained from the San Diego Blood Bank were cultured at 2 × 10 in RPMI + GlutaMAX (Gibco, Cat. No. 61870036) containing 100 U penicillin, 100 μg streptomycin (Gibco, Cat. No. 10378016), and 10% FBS (RPMI). 5 cells / well into a 96-well flat-bottom plate (GenClone, Cat. No. 25-109) or 1.7 x 10 5Cells / well were seeded into round-bottom plates (GenClone, Cat. No. 25-221). Test antibodies were added to wells at 4x the final concentration in a volume of 50 μL. 50 μL of recombinant human IL-12 (R&D systems, Cat. No. 219-IL-005) was then added at 4 ng / mL, followed by 50 μL of recombinant human IL-18 (Sino Biological, Cat. No. 10119-HNCE) at 8 ng / mL, for final concentrations of 1 ng / mL and 2 ng / mL, respectively. For assays using cyno PBMCs (iQ Biosciences, Cat. No. IQB-MnPB102), cells were plated at 1.7 x 10 cells / well in round-bottom plates. 5 Cells were seeded at 1000 x g / well. Recombinant cynomolgus IL-12 (R&D Systems, Cat. No. 10215-CL) was added at a final concentration of 1 ng / mL, and recombinant rhesus IL-18 (R&D Systems, Cat. No. 2548-RM-025 / CF [note that the amino acid sequences of rhesus and cynomolgus IL-18 are identical]) was added at a final concentration of 2 ng / mL. All dilutions were performed in RPMIc. Cells were incubated at 37°C, 5% CO2 for 48 hours. At 48 hours, 50 μL aliquots of supernatant were removed from each well and analyzed for the presence of IFNγ using a DuoSet ELISA kit (R&D Systems, catalog number DY285B for human IFNγ and catalog number DY961 for primate IFNγ) on a Nunc MaxiSorp Flat-Bottom Plate (Invitrogen, catalog number 44-2404-21) according to the manufacturer's instructions. CCL2 release was measured using a DuoSet ELISA kit (R&D Systems, catalog number DY279 for human CCL2) on a Nunc MaxiSorp Flat-Bottom Plate (Invitrogen, catalog number 44-2404-21) according to the manufacturer's instructions. Absorbance was measured at 450 nm using a Spectramax iD5 plate reader, and data was analyzed using GraphPad Prism software.

[0216] For experiments in which reactions were performed in the presence of pre-complexed hIL-18 / hIL-18BP, the design was as follows: 80 ng / mL recombinant human or cynomolgus IL-18 was incubated with recombinant human (SinoBiological, catalog no. 10357-H08H) or cynomolgus (produced in-house) IL-18BP, respectively, at 400 ng / mL at room temperature for 30 minutes. Serial dilutions of mAb were added to a 96-well round-bottom plate at 4x concentration in 50 μL / well. 50 μL / well of IL-18-IL-18BP complex was added to each well containing mAb and incubated for 1 hour at 37°C. After 1 hour, 50 μL / well of recombinant human or cynomolgus IL-12 was added at 4 ng / mL, and 50 μL of PBMCs were added at 2x10 6 cells / mL to each well to give a final concentration of 20 ng / mL IL-18, 100 ng / mL IL-18BP, 1 ng / mL IL-12, and 1 x 10 5 The final concentration of PBMCs was 100 μg / mL. All dilutions were performed in RPMIc. Control wells contained IL-12 + IL-18 alone or IL-12 + IL-18 + IL-18BP alone. Cells were incubated at 37°C, 5% CO2 for 48 hours, and supernatants were collected and analyzed for the presence of IFNγ and CCL2.

[0217] NK cell assay for human IL-18 activity. IL-18 activity was assessed using purified NK cells and precomplexed hIL-18 / hIL-18BP. 10 ng / mL recombinant human IL-18 (4x final concentration) was incubated with 50 ng / mL recombinant human IL-18BP (4x final concentration) for 30 minutes at room temperature. NK cells were purified from fresh PBMCs using the MojoSort Human NK Cell Isolation Kit (BioLegend, catalog no. 480054) according to the manufacturer's instructions. NK purity was assessed by flow cytometry and was often greater than 90%. 1.7 x 10 cells were cultured per well of a 96-well flat-bottom plate. 5NK cells were added in 50 μL of RPMIc. Pre-complexed hIL-18 / hIL-18BP was added at 50 μL / well, followed by 50 μL of recombinant human IL-12 at 4 ng / mL (for a final concentration of 1 ng / mL) and incubated at 37°C for 30 minutes. Serial dilutions of test antibodies were added at 50 μL / well at 4x concentrations. Plates were incubated at 37°C, 5% CO2 for 24 hours. Supernatants were collected and analyzed for the presence of IFNγ by ELISA as described above.

[0218] result Isolation of monoclonal antibody sequences from immunized mice. DiversimAb™ and DivergimAb™ mice (Abveris) were immunized with human and cynomolgus IL-18BP. Titers against human, cynomolgus, and mouse IL-18BP were measured by ELISA. Mice with high titers were selected for isolation of antibody-secreting B cells using the Berkeley Lights Beacon Optofluidic System (Mullen, 2021). Screening identified single cells secreting antibodies that cross-reacted with human and cynomolgus IL-18BP, some of which also bound to mouse IL-18BP. Furthermore, a novel screen was designed to identify cells secreting antibodies that failed to recognize an active-site-blocked form of IL-18BP, termed "hypo-IL-18." This enabled the screening to identify putative ligand-blocking antibodies.

[0219] Because IL-18 exhibits high affinity for binding proteins, with a KD of less than 1 nM (Kim et al., PNAS 97(3):1190-1195, 2000; Kimura et al., Allergol Int. 57(4):367-76, 2008), antagonizing the IL-18 / IL-18BP interaction may not be possible with mAbs with affinities in the nM range, such as those typically isolated from antigen-specific B cells. Therefore, to evaluate the potential blocking ability of newly identified mAb candidates, we created a novel chimeric protein, "low IL-18." Low IL-18 consists of human or mouse IL-18 tethered to its respective IL-18BP, separated by a flexible linker peptide (schematically depicted in Figure 1A-1B). The sequence of low IL-18 is provided below in Table S1. [Table 4]

[0220] The rationale for generating this molecule is that the tethered ligand cannot dissociate, blocking the active site of IL-18BP. Thus, mAbs that recognize the active binding site of BP are sterically hindered from recognizing low-density IL-18, whereas most unblocked antibodies can bind equally well to IL-18BP and low-density IL-18. The design also included a TEV protease cleavage site, allowing IL-18 to be separated from its binding protein.

[0221] The design of a low-IL-18 construct, linking the C-terminus of IL-18 to the N-terminus of IL-18BP, was based on the crystal structure (Protein Data Bank [PDB] structure 3F62). This structure contains human IL-18 complexed with ectromelia virus IL-18BP. Because orthopoxviruses, including Ectromelia, encode functional IL-18BP homologs that exhibit 17–34% amino acid identity with the mammalian ortholog IL-18BP (Calderara, 2001), the crystal structure of 3F62 was considered a guide for the design of the low-IL-18 construct. Analysis of the crystal structure suggested that the C-terminus of IL-18 is relatively close to the N-terminus of the binding protein, making a fusion protein feasible. Thus, certain embodiments include reduced IL-18 fusion proteins comprising, from N- to C-terminal, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and IL-18BP, wherein the IL-18 portion of the fusion protein is linked to the IL-18BP portion of the fusion protein and sterically blocks the IL-18-binding portion of the IL-18BP portion of the fusion protein. In certain embodiments, the reduced IL-18 fusion protein comprises, consists of, or consists essentially of an amino acid sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence in Table S1. Nucleic acid molecules encoding reduced IL-18 fusion proteins are also included.

[0222] Confirmation of activity of re-expressed mAbs, analysis of binding kinetics, blocking capacity, and biological activity. The antibody sequences were derived from single B cell screening on the Beacon instrument. To confirm activity, the antibodies were recombinantly re-expressed using isolated mouse variable region sequences fused to the human IgG1 kappa constant region. The antibodies were expressed in HEK293 cells and tested from cell culture supernatants or purified for further analysis.

[0223] Initial characterization used BLI to test binding affinity to human, cynomolgus monkey, and mouse IL-18BP. Additionally, mAbs were tested for binding to low levels of IL-18 to identify mAbs with the highest potential for blocking. The mAbs were also scored for their expression level and whether the sequence was considered problematic. Examples of problematic sequences are polytyrosine sequences in CDR3 or low levels of somatic hypermutation, indicating that the antibody had not undergone significant in vivo maturation. The results are shown in Table E1. [Table 5]

[0224] The antibodies of interest were also tested for activity in a cell-based reporter assay. This assay uses a cell line engineered to secrete alkaline phosphatase in response to IL-18 signaling, which is easily identified. IL-18 induction of alkaline phosphatase is inhibited by the addition of IL-18BP. This inhibition is alleviated by the addition of antibodies against IL-18BP, if these antibodies are capable of blocking IL-18BP / IL-18 binding. As shown in Table E2, some antibodies were able to block the IL-18 / IL-18BP interaction and induce IL-18-induced responses. Antibodies that generated activity in this assay were consistent with antibodies that were unable to bind low levels of IL-18, validating the screening method used to isolate the antibodies of interest. [Table 6]

[0225] The antibodies were then tested for biological activity in KG-1 cells. KG-1 is a human bone marrow-derived macrophage cell line that responds to IL-18 by producing IFNγ. Addition of IL-18BP blocks the ability of IL-18 to induce IFNγ expression, suppressing the response. Further addition of an anti-IL-18BP neutralizing antibody binds to IL-18BP and prevents its interaction with IL-18, liberating IL-18 and inducing IFNγ. The antibodies demonstrated the ability to liberate IL-18 from IL-18BP inhibition in this assay, as shown in Table E3. [Table 7]

[0226] mAb optimization Two mAbs were prioritized for humanization and optimization: SA04a and SA44a (see Table E4). [Table 8]

[0227] Humanization of SA04a. The heavy chain CDRs of SA04a were grafted onto three different human variable regions: IGHV7-4-1*02, IGHV1-3*04, and IGHV3-23, and the light chain CDRs were grafted onto hIGKV1-33*01, which contains the framework backmutations P44V, F71Y, and Y87F. The grafted heavy chain was grafted onto IGHV7-4-1*02, which contains the framework backmutations G26V, V37L, L45F, and Y91F. The binding characteristics of the resulting humanized mAb (SA50a) are shown in Table E5 below. [Table 9]

[0228] Humanization of SA44a. The HC and LC CDRs from the SA44a antibody obtained from DivergimAb™ were grafted onto the human germline variable regions IgHV4-30-4*01 and IGKV3-11*01, respectively. The grafting process significantly reduced binding affinity. Therefore, to optimize affinity, two back mutations (G27Y in FW1 and V71R in FW3) were reintroduced into the HC and one back mutation (L46P in FW2) was reintroduced into the LC. As shown in Table E6 below, the resulting humanized mAb (SA301a) restored full activity and superior expression compared to its original murine parent (SA44a). [Table 10]

[0229] SA301a maturation. To affinity-maturate mAb SA301a, a library of mAb variants centered on the HC and LC CDRs was prepared. To do this, amino acids in the CDRs were then replaced with up to 17 amino acid substitutions. To avoid introducing potential undesirable sequence defects, cysteines and tryptophans were not included in the library, and parent amino acids already in place were not included in the screening. Upon generation, the variants were first screened by BLI at a single concentration of human and cynomolgus IL-18BP to determine whether there were any improvements in on- or off-rates compared to the parent antibody, SA301a. Screening identified a variety of single-point variants that indeed had demonstrable improved binding kinetics. In some cases, but not all, individual mutations could be recombined to further improve binding affinity for the target antigen.

[0230] The initial screen also identified a subset of amino acid substitutions that specifically improved binding to both human and cynomolgus IL-18BP. Consequently, these changes were recombined in various combinations for further analysis.

[0231] Table E7 shows variants that led to improved binding to human and cynomolgus IL-18BP. The (N) values ​​in brackets indicate how many data points from independently performed experiments were included in the average, and ± indicates SEM. NT = not tested. ND = not determined. If a mAb was tested only twice, both values ​​are provided. Cynomolgus / human provides the ratio difference between the two species. [Table 11-1] [Table 11-2]

[0232] Further characterization of high affinity mAbs against human and cynomolgus IL-18BP The mAbs with high affinity for human and cynomolgus IL-18BP were further characterized to identify those suitable for further development. This analysis included evaluation of antibody binding properties (e.g., blocking IL-18BP-mediated neutralization of IL-18BP signaling; human IgG1 / kappa; cross-reacting with cynomolgus IL-18BP; high affinity for human IL-18BP with a KD lower than the KD of IL-18 and IL-18BP; no detectable nonspecific binding or binding to homologous proteins, etc.), functional properties (e.g., blocking IL-18 binding to IL-18BP; neutralizing IL-18BP, thus antagonizing IL-18-mediated suppression of IFNγ induction, etc.), and developability.

[0233] Functional activity. The ability of the subject mAbs to inhibit neutralization of IL-18 by IL-18BP was assessed in a number of different bioassays. These assays included activity in PBMC assays for IL-18 activity in two different formats. Experimental details for these assays are provided in the Materials and Methods section above.

[0234] IL-18, in conjunction with IL-12, induces IFNγ production by multiple cell types in PBMCs, including NK cells and T cells. IL-18BP is endogenously produced by PBMCs and upregulated by IFNγ, providing a negative feedback loop that suppresses IFNγ responses. Addition of an anti-IL-18BP neutralizing antibody binds to endogenous IL-18BP, inhibiting this negative feedback loop and inducing IFNγ production. The ability of a set of mAbs to compete with IL-18 for binding to IL-18BP and induce IL-18-mediated IFNγ was tested in vitro in healthy donor PBMCs. The concentration of IL-18 used in the assay was only sufficient to generate a low-level IFNγ response in PBMCs, likely due to inhibition by endogenous IL-18BP.

[0235] As shown in Table E8 below and Figures 2A-2G, addition of mAb relieved this inhibition and resulted in a dose-dependent increase in IFNγ production. Antibody potency correlated broadly, but not exclusively, with their binding affinity for IL-18BP.

[0236] Additionally, a second assay format was performed using human PBMCs. In this format, IL-18 was precomplexed with IL-18BP before addition to PBMCs. This assay tests the ability of antibodies to release IL-18 from pre-existing complexes with IL-18BP. As shown in Figures 3A-3G, activity was also observed in this assay, and the potency of IFNγ induction again correlated in most cases with the affinity of the highest affinity mAb. EC50 values ​​are provided in Table E8 below. [Table 12]

[0237] Anti-human IL-18BP antibodies were also able to induce IL-18-induced production of additional cytokines, such as CCL2, from PBMCs. The antibodies were tested in PBMC Assay 1 (the endogenous IL-18BP assay described above), and CCL2 release was determined by ELISA. Addition of anti-IL-18BP mAb increased CCL2 production by PBMCs (Figure 4).

[0238] The activity of the anti-IL-18BP mAb was also tested using a cynomolgus monkey PBMC assay. Both assay formats described above for human PBMC were performed using PBMC isolated from cynomolgus monkey blood. The antibody demonstrated activity in both formats, as shown in Figures 5A-5B.

[0239] Activity was also tested in an NK cell-specific assay. In this assay, purified human NK cells were incubated with precomplexed IL-18:IL-18BP, with or without the test antibody. As shown in Figure 6, the test antibody was able to disrupt the IL-18:IL-18BP complex and induce IFNγ secretion from NK cells.

[0240] Library Screening Data The binding properties were tested for variants of the parent SA301a and SA302a antibodies.

[0241] Heavy chain library screening. Octet production kinetic values ​​measured >50 nM are listed as "inactive." Positions resulting in ≥1.5-2.0-fold improvements for humans and cynomolgus monkeys (at the time of testing) were considered for further analysis. Values ​​presented for each species are apparent KDs; these were calculated from single binding concentrations collected in a semi-high-throughput format. Samples were compared to the parental background from which they were generated (normal font if the parent was SA301a; italic font if the parent was SA302a, which contains HC-C34A). [Table 13] [Table 14] [Table 15] [Table 16] [Table 17-1] [Table 17-2] [Table 18]

[0242] Light chain library screening. Octet production kinetic values ​​measured >50 nM are listed as "inactive." Positions resulting in ≥1.5-2.0-fold improvements for both human and cynomolgus monkeys were considered for further analysis. Values ​​presented for each species are apparent KDs; these were calculated from single binding concentrations collected in a semi-high-throughput format. Samples are compared to the parental background from which they were generated; i.e., font is normal if the parent is SA301a, and italic if the parent is SA326a (HC-C34A x LC-Q90L). [Table 19-1] [Table 19-2] [Table 20] [Table 21-1] [Table 21-2]

[0243] Epitope mapping The epitope of mAb SA338a on IL-18BP was determined by a crosslinking / high-resolution mass spectrometry method developed by CovalX AG (Pimenova et al., 2008, J. Mass Spectrometry 43:185). Briefly, human IL-18BP was conjugated to SA338a and crosslinked with a heterobifunctional linker. The resulting complex was digested with five different proteases (trypsin, chymotrypsin, ASP-N, elastase, and thermolysin), and the resulting peptides, whether crosslinked or not, were analyzed by high-resolution mass spectrometry.

[0244] The results indicated that SA338a recognizes a conformational epitope containing the following residues in IL-18BP. Furthermore, residues on IL-18BP that interact with IL-18 were similarly mapped using the same technique. The results demonstrate that four residues on IL-18BP are involved in IL-18 recognition and are also recognized by SA338a. These results support evidence that the antibody is a functional blocking antibody (S75, H79, T116, and S119). SA338a interacts with the following residues on IL-18BP: T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119. The residues on IL-18BP that interact with IL-18 have been identified as follows: R61, Y69, S75, H79, T116, S119, and R131. The sequence of mature IL-18BP is shown below. Residues that interact with SA338a and IL-18 are highlighted as shown. TIFF2026503273000038.tif36164

[0245] Residues known to interact with IL-18 are highlighted in bold and underlined. Residues that form the epitope for SA338a are highlighted in italics and bold. Residues S75, H79, T116, and S119 are recognized by both IL-18 and SA338a and are shown in bold, italics, and underlined.

Claims

1. An antibody specific to interleukin-18 binding protein (IL-18BP), which interferes with the binding of IL-18 to IL-18BP.

2. The antibody of claim 1, wherein the antibody binds to a conformational epitope of IL-18BP.

3. The antibody of claim 2, wherein the antibody binds to two or more of amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO:

372.

4. The antibody of claim 2, wherein the antibody binds to amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO:

372.

5. The antibody of claim 1, wherein the antibody binds to a linear epitope of IL-18BP.

6. The antibody according to any one of claims 1 to 5, which binds to the binding interface between IL-18 and mature IL-18BP.

7. The antibody of claim 6, wherein the antibody binds to amino acid residues S75, H79, T116, and S119 of SEQ ID NO:

372.

8. The antibody according to any one of claims 1 to 7, which binds to human IL-18BP and cynomolgus monkey IL-18BP, but does not bind to mouse IL-18BP.

9. The antibody according to any one of claims 1 to 7, which binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

10. The antibody is a fusion protein that binds to IL-18 and IL-18BP with a high affinity (K Ｄ 10. The antibody of any one of claims 1 to 9, which binds to IL-18BP with a binding affinity of greater than about 650 pM, optionally between about 1 pm and about 650 pm, or about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM or less.

11. The antibody according to any one of claims 1 to 10, which is an IL-18BP antagonist that antagonizes the binding activity between IL-18BP and IL-18.

12. The antibody of claim 11, wherein the antibody blocks the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signaling, including the induction of IFN-gamma, CXCL10 and / or TNFα.

13. 13. The antibody of any one of claims 1 to 12, comprising an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or hybrids and / or variants thereof.

14. 14. The antibody of claim 13, comprising an IgG Fc domain, optionally an IgG1 or IgG3 Fc domain, that has enhanced effector function in humans.

15. 14. The antibody of claim 13, comprising an IgG Fc domain with reduced effector function in humans, optionally an IgG2 or IgG4 Fc domain.

16. The antibody according to any one of claims 1 to 15, wherein the antibody is a monoclonal antibody.

17. The antibody according to any one of claims 1 to 16, wherein the antibody is a humanized antibody.

18. The antibody of any one of claims 1 to 17, wherein the antibody is an scFv.

19. The antibody a. V of a complementarity determining region selected from Table A1 Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The heavy chain variable region (V) containing the CDR3 sequence Ｈ ), and variants thereof that specifically bind to IL-18BP; and b. V of a complementarity determining region selected from Table A1 Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ The light chain variable region (V) containing the CDR3 sequence Ｌ 19. The antibody according to any one of claims 1 to 18, comprising: a IL-18BP-specifically binding IL-18BP; and a variant thereof.

20. a. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 1-3, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 4-6, respectively; b. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 7-9, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 10-12, respectively; c. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 13-15, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 16-18, respectively; d. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 19-21, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 22-24, respectively; e. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 25-27, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 28-30, respectively; f. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 31-33, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 34-36, respectively; g. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 37-39, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 40-42, respectively; h. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 43-45, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 46-48, respectively; i. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 49-51, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 52-54, respectively; j. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 55-57, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 58-60, respectively; k. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 61-63, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 64-66, respectively; l. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 67-69, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 70-72, respectively; m. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 73-75, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 76-78, respectively; n. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 79-81, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 82-84, respectively; o. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 85-87, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 88-90, respectively; p. V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 91-93, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 94-96, respectively; q. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 97-99, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 100-102, respectively; r. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 103-105, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 106-108, respectively; s. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 109-111, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; t. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 115-117, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; u. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 121-123, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 124-126, respectively; v. The above V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 127-129, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 130-132, respectively; w. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 133-135, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 136-138, respectively; x. The above V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 139-141, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 142-144, respectively; y. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 145-147, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 148-150, respectively; z. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 151-153, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 154-156, respectively; aa. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 157-159, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 160-162, respectively; bb. V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 163-165, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; cc. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 169-171, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 172-174, respectively; dd. The above V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 175-177, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 178-180, respectively; ee. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 181-183, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 184-186, respectively; ff. The above V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 187-189, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences include SEQ ID NOs: 190-192, respectively; gg. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 193-195, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; hh. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 199-201, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; ii. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 205-207, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs:208-210, respectively; jj. The above V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 211-213, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs:214-216, respectively; kk. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 217-219, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; ll. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 223-225, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; mm. Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 229-231, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 232-234, respectively; nn. Said V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 235-237, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; oo. The above V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 241-243, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 244-246, respectively; pp. V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 250-252, respectively; qq. The V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 253-255, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 256-258, respectively; or rr. V Ｈ CDR1 sequence, V Ｈ CDR2 sequence, and V Ｈ The CDR3 sequences comprise SEQ ID NOs: 259-261, respectively, and Ｌ CDR1 sequence, V Ｌ CDR2 sequence, and V Ｌ The antibody of claim 19, wherein the CDR3 sequences comprise SEQ ID NOs: 262 to 264, respectively.

21. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally said V Ｈ 21. The antibody of any one of claims 1 to 20, wherein said antibody has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

22. The V Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally said V Ｌ 22. The antibody of any one of claims 1 to 21, wherein said antibody has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

23. a. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 265, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 266; b. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 267, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 268; c. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 269, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 270; d. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 271, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 272; e. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 273, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 274; f. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 275, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 276; g. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 277, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 278; h. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 279, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 280; i. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 281, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 282; j. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 283, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 284; k. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 285, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 286; l. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 287, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 288; m. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 289, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 290; n. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 291, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 292; o. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 293, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 294; p. V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 295, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 296; q. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 297, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 298; r. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 299, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 300; s. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 301, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 302; t. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 303, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 304; u. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 305, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 306; v. The above V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 307, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 308; w. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 309, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 310; x. The above V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 311, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 312; y. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 313, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 314; z. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 315, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 316; aa. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 317, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 318; bb. V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 319, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 320; cc. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 321, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 322; dd. The above V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 323, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 324; ee. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 325, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 326; ff. The above V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 327, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 328; gg. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 329, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 330; hh. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 331, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 332; ii. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 333, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 334; jj. The above V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 335, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 336; kk. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 337, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 338; ll. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 339, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 340; mm. Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 341, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 342; nn. Said V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 343, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 344; oo. The above V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 345, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 346; pp. V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 347, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 348; qq. The V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 349, and Ｌ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 350; or rr. V Ｈ comprises a sequence at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 351, and Ｌ 23. The antibody of any one of claims 1 to 22, comprising a sequence that is at least 70, 75, 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:

352.

24. An isolated polynucleotide encoding the isolated anti-IL-18BP antibody of any one of claims 1 to 23, an expression vector comprising said isolated polynucleotide, or an isolated host cell comprising said vector.

25. A pharmaceutical composition comprising the anti-IL-18BP antibody according to any one of claims 1 to 23 and a pharmaceutically acceptable carrier.

26. 26. The pharmaceutical composition of claim 25, wherein the composition is a sterile injectable solution, optionally suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

27. A method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of claims 25-26.

28. 28. The method of claim 27, wherein the disease or condition is a cancer, or a tumor, or a proliferative disease or disorder, optionally a proliferative disease or disorder selected from lymphoproliferative disorders, myeloproliferative disorders, proliferative enterocolitis, proliferative diabetic retinopathy, and proliferative kidney disease.

29. 29. The method of claim 28, wherein the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18.

30. The cancers include bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), hepatocellular carcinoma (liver cell carcinoma), sarcoma, B-cell malignancies, 30. The method of claim 28 or 29, wherein the tumor is selected from one or more of: breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, urothelial cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer.

31. The method according to any one of claims 27 to 30, comprising administering the pharmaceutical composition according to claim 25 or 26 in combination with IL-18.

32. 32. The method of any one of claims 28 to 31, comprising administering the pharmaceutical composition of claim 25 or 26 in combination with an immune checkpoint modulator selected from an antagonist of an inhibitory immune checkpoint molecule, and an agonist of a stimulatory immune checkpoint molecule.

33. 33. The method of claim 32, wherein said immune checkpoint modulator is a polypeptide, optionally an antibody, or a ligand, or a small molecule.

34. The inhibitory immune checkpoint molecules include Programmed Death-Ligand 1 (PD-L1), Programmed Death 1 (PD-1), Programmed Death-Ligand 2 (PD-L2), Cytotoxic T-Lymphocyte-Associated protein 4 (CTLA-4), Indoleamine 2,3-dioxygenase (IDO), Tryptophan 2,3-dioxygenase (TDO), T-cell Immunoglobulin domain and Mucin domain 3 (TIM-3), Lymphocyte Activation Regulatory Factor (LIF-FRF) and Lymphocyte Activation Factor (LIF-FRF). The method of claim 32 or 33, wherein the target gene is selected from one or more of Gene-3 (LAG-3), V-domain Ig suppressor of T cell activation (VISTA), B and T lymphocyte attenuator (BTLA), CD160, Herpes Virus Entry Mediator (HVEM), and T-cell immunoreceptor with Ig and ITIM domains (TIGIT).

35. a. the antagonist is an antagonist of PD-L1 and / or PD-L2 optionally chosen from one or more of an antibody or small molecule that specifically binds to PD-L1 and / or PD-L2, atezolizumab (MPDL3280A), avelumab (MSB0010718C), and durvalumab (MEDI4736), and optionally the cancer is chosen from one or more of colorectal cancer, melanoma, breast cancer, non-small cell lung cancer, bladder cancer, and renal cell carcinoma; b. the antagonist is a PD-1 antagonist optionally chosen from one or more of an antibody or small molecule that specifically binds to PD-1, nivolumab, pembrolizumab, MK-3475, AMP-224, AMP-514PDR001, and pidilizumab, optionally wherein the PD-1 antagonist is nivolumab, and the cancer is optionally chosen from one or more of Hodgkin's lymphoma, melanoma, non-small cell lung cancer, hepatocellular carcinoma, renal cell carcinoma, and ovarian cancer; c. the PD-1 antagonist is pembrolizumab and the cancer is optionally selected from one or more of melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, and urothelial cancer; d. the antagonist is a CTLA-4 antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to CTLA-4, ipilimumab, tremelimumab, and optionally the cancer is selected from melanoma, prostate cancer, lung cancer, and bladder cancer; e. the antagonist is an IDO antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to IDO, indoximod (NLG-8189), 1-methyl-1-tryptophan (1MT), β-carboline (norharman; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat, and the cancer is optionally selected from one or more of metastatic breast cancer and brain cancer, optionally glioblastoma multiforme, glioma, gliosarcoma, or malignant brain tumor; f. the antagonist is a TDO antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to TDO, 680C91, and LM10; g. the antagonist is a TIM-3 antagonist optionally selected from one or more of an antibody or a small molecule that specifically binds to TIM-3; h. the antagonist is a LAG-3 antagonist optionally selected from one or more of an antibody or small molecule that specifically binds to LAG-3, and BMS-986016; i. the antagonist is a VISTA antagonist optionally selected from one or more of an antibody or a small molecule that specifically binds to VISTA; j. the antagonist is an antagonist of BTLA, CD160 and / or HVEM, optionally selected from one or more of an antibody or a small molecule that specifically binds to BTLA, CD160 and / or HVEM; or k) The method of claim 34, wherein the antagonist is a TIGI antagonist optionally selected from one or more of an antibody or a small molecule that specifically binds to TIGIT.

36. 34. The method of claim 32 or 33, wherein the stimulatory immune checkpoint molecule is selected from one or more of OX40, CD40, Glucocorticoid-Induced TNFR Family Related Gene (GITR), CD137 (4-1BB), CD27, CD28, CD226, and Herpes Virus Entry Mediator (HVEM).

37. a. the agonist is an OX40 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to OX40, OX86, Fc-OX40L, and GSK3174998; b. the agonist is a CD40 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD40, CP-870,893, dacetuzumab, Chi Lab 7 / 4, ADC-1013, and rhCD40L, and the cancer is optionally selected from one or more of melanoma, pancreatic cancer, mesothelioma, and hematological cancer, optionally a lymphoma, e.g., non-Hodgkin's lymphoma; c. the agonist is a GITR agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to GITR, INCAGN01876, DTA-1, and MEDI1873; d. the agonist is a CD137 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD137, utomilumab, and 4-1BB ligand; e. the agonist is a CD27 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD27, varlilumab, and CDX-1127 (1F5); f. the agonist is a CD28 agonist optionally selected from one or more of an antibody or small molecule or ligand that specifically binds to CD28, and TAB08; and / or g. The method of claim 36, wherein the agonist is an HVEM agonist, optionally selected from one or more of an antibody or a small molecule or a ligand that specifically binds to HVEM.

38. The method according to any one of claims 28 to 37, comprising administering the pharmaceutical composition according to claim 25 or 26 in combination with at least one chemotherapeutic agent.

39. 39. The method of claim 38, wherein the at least one chemotherapeutic agent is selected from one or more of an alkylating agent, an antimetabolite, a cytotoxic antibiotic, a topoisomerase inhibitor (type 1 or type II), and an anti-microtubule agent.

40. a. said alkylating agent is selected from one or more of nitrogen mustards (optionally mechlorethamine, cyclophosphamide, mustine, melphalan, chlorambucil, ifosfamide, and busulfan), nitrosoureas (optionally N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, and streptozotocin), tetrazines (optionally dacarbazine, mitozolomide, and temozolomide), aziridines (optionally thiotepa, mitomycin, and diaziquone (AZQ)), cisplatin and its derivatives (optionally carboplatin and oxaplatin), and non-classical alkylating agents (optionally procarbazine and hexamethylmelamine); b. said antimetabolite is selected from one or more of antifolates (optionally methotrexate and pemetrexed), fluoropyrimidines (optionally 5-fluorouracil and capecitabine), deoxynucleoside analogs (optionally ancitabine, enocitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, fludarabine, and pentostatin), and thiopurines (optionally thioguanine and mercaptopurine); c. the cytotoxic antibiotic is selected from one or more of anthracyclines (optionally doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin, and mitoxantrone), bleomycin, mitomycin C, mitoxantrone, and actinomycin; d. the topoisomerase inhibitor is selected from one or more of camptothecin, irinotecan, topotecan, etoposide, doxorubicin, mitoxantrone, teniposide, novobiocin, mervalone, and aclarubicin; and / or e) The method of claim 39, wherein the anti-microtubule agent is selected from one or more of taxanes (optionally paclitaxel and docetaxel) and vinca alkaloids (optionally vinblastine, vincristine, vindesine, vinorelbine).

41. 28. The method of claim 27, wherein the disease or condition is myelodysplastic syndrome (MDS).

42. 28. The method of claim 27, wherein the disease or condition is an infectious disease.

43. 43. The method of claim 42, wherein the infectious disease is selected from viral, bacterial, fungal (optionally yeast), and protozoal infections.

44. The method according to any one of claims 41 to 43, comprising administering the pharmaceutical composition according to claim 25 or 26 in combination with IL-18.

45. 1. A method for screening anti-IL-18BP antibodies for their ability to block or inhibit binding between IL-18 and IL-18BP, comprising: a. determining the binding affinity of the antibody to (i) IL-18BP alone, and (ii) an IL-18-reduced fusion protein, the IL-18-reduced fusion protein comprising IL-18 fused to IL-18BP via a flexible linker (and an optional protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein is bound to the IL-18BP portion of the fusion protein so as to sterically block the IL-18 binding site of the IL-18BP portion of the fusion protein; b. comparing the binding affinity of (i) with the binding affinity of (ii); and c) identifying or selecting the antibody as being capable of blocking or inhibiting the binding between IL-18 and IL-18BP if the binding affinity of (i) is significantly stronger than the binding affinity of (ii).

46. The method of claim 45, wherein the IL-18 and IL-18BP are mouse IL-18 and IL-18BP.

47. The method of claim 45, wherein the IL-18 and IL-18BP are human IL-18 and IL-18BP.

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