Activation of t cells with mutant g-csf r

Abstract

Provided herein are particles or compositions comprising one or more nucleic acid molecules encoding a chimeric antigen receptor (CAR) and / or a polypeptide comprising i) a ligand binding extracellular domain (ECD), ii) an immune cell activating intracellular domain (ICD); and iii) a transmembrane domain linking the ECD and the ICD, polypeptides comprising the domains, and nucleic acid molecules encoding the polypeptides. Also provided herein are compositions comprising the nucleic acid molecules and methods of using the compositions. Also provided herein are viral particles comprising the nucleic acid molecules.

Description

[0001] This application is a divisional application of the invention patent application filed on June 13, 2024, with application number 202480046363.0 and invention title "Activation of T cells using mutant G-CSFR".

[0002] Related applications

[0003] This application claims the benefits of U.S. Provisional Application Serial No. 63 / 508,055, filed June 14, 2023, and U.S. Provisional Application Serial No. 63 / 510,745, filed June 28, 2023, each of which is incorporated herein by reference in its entirety.

[0004] Reference sequence list submitted electronically

[0005] This application includes a sequence list, which is submitted electronically in XML format and incorporated herein by reference in its entirety. The XML copy created on June 5, 2024, is named “INH-023WO_SL.xml” and has a size of 146,320 bytes. Technical Field

[0006] The embodiments provided herein relate to mutant G-CSFR constructs and methods of using these constructs. The embodiments provided herein also relate to viral particles comprising a heterologous viral glycoprotein, a targeting moiety, and a nucleic acid molecule encoding a heterologous molecule of interest. In some embodiments, the nucleic acid molecule encodes a chimeric antigen receptor and a mutant G-CSFR. The embodiments provided herein further relate to compositions comprising the viral particles provided herein and methods of using these compositions. Background Technology

[0007] Chimeric antigen receptor T-cell (CAR-T) cell therapy has revolutionized anti-tumor treatment, particularly for hematologic malignancies. However, significant limitations such as high cost, long manufacturing cycles, and safety concerns remain and hinder the widespread applicability of this technology. In vivo CAR-T is based on the concept that CAR molecules can be delivered in vivo to target T cells using a T-cell targeting carrier. Traditional CAR-T methods require the isolation and proliferation of individualized T cells before infusion back into the patient after CAR transduction. Therefore, a potential drawback of in vivo CAR-T is the assumption that CAR-containing T cells lack proliferation, which may limit the efficacy of the method. Embodiments of the present invention address these and other needs. Summary of the Invention

[0008] In some embodiments, particles or compositions are provided. In some embodiments, the particles or compositions comprise one or more nucleic acid molecules encoding: a) a chimeric antigen receptor (CAR); and b) a polypeptide comprising: i) a ligand-binding extracellular domain (ECD); ii) an immune cell-activating intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD.

[0009] In some embodiments, the immune cell activation ICD comprises an ICD of hGCSF-R (human granulocyte colony-stimulating factor receptor) or a mutant or fragment thereof. In some embodiments, the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111, or an amino acid sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

[0010] In some embodiments, the ECD is a GCSF-R ECD. In some embodiments, the GCSF-R ECD is a human GCSF-R ECD. In some embodiments, the human GCSF-R ECD contains the amino acid sequence of SEQ ID NO: 107 or SEQ ID NO: 108.

[0011] In some embodiments, the ECD is a human growth hormone receptor (hGH-R) ECD or a mutant or fragment thereof. In some embodiments, the hGH-R ECD comprises the amino acid sequence of SEQ ID NO: 114 or SEQ ID NO: 115, or an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

[0012] In some embodiments, the peptide comprises: an ECD as a GCSFR-ECD or a variant thereof (e.g., as shown herein); a transmembrane domain as a GCSF-R transmembrane domain or a variant thereof (e.g., as shown herein); and an immune-activated ICD as a GCSF-R ICD or a variant thereof (e.g., as shown herein).

[0013] In some embodiments, the peptide comprises: an ECD as hGH-R-ECD or a variant thereof (e.g., as shown herein); a transmembrane domain as a GCSF-R transmembrane domain or a variant thereof (e.g., as shown herein); and an immune-activated ICD as a GCSF-R ICD or a variant thereof (e.g., as shown herein).

[0014] In some embodiments, the peptide comprises: an ECD as hGH-R-ECD or a variant thereof (e.g., as shown herein); a transmembrane domain as gp130 transmembrane domain or a variant thereof (e.g., as shown herein); and an immune-activated ICD comprising gp130 ICD or a variant thereof (e.g., as shown herein) and IL-2Rβ ICD or a variant thereof (e.g., as shown herein).

[0015] In some embodiments, the peptide comprises: an ECD as hGH-R-ECD or a variant thereof (e.g., as shown herein); a transmembrane domain as a GCSF-R transmembrane domain or a variant thereof (e.g., as shown herein); and an immune-activated ICD comprising a GCSF-R ICD or a variant thereof (e.g., as shown herein) and an IL-7Rα ICD or a variant thereof (e.g., as shown herein).

[0016] In some embodiments, the polypeptide comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124, or a variant thereof. In some embodiments, the variant comprises a polypeptide having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124.

[0017] In some embodiments, a nucleic acid molecule is provided that encodes a polypeptide comprising i) a ligand-binding extracellular domain (ECD) as provided herein; ii) an immune cell-activating intracellular domain (ICD) as provided herein; and iii) a transmembrane domain linking the ECD to the ICD as provided herein.

[0018] In some embodiments, the nucleic acid molecule further encodes a chimeric antigen receptor (CAR). In some embodiments, the CAR is as provided herein. In some embodiments, the CAR is a CD20 CAR as provided herein. In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity with SEQ ID NO: 99, at least 90% identity with SEQ ID NO: 99, at least 95% identity with SEQ ID NO: 99, at least 99% identity with SEQ ID NO: 99, or at least 100% identity with SEQ ID NO: 99. In some embodiments, the nucleic acid sequence encoding the polypeptide and the nucleic acid sequence encoding the CAR are separated by a nucleic acid sequence encoding a cleavable peptide sequence (such as the 2A sequence).

[0019] In some embodiments, a polypeptide is provided. In some embodiments, the polypeptide comprises i) a ligand-binding extracellular domain (ECD) as provided herein; ii) an immune cell-activating intracellular domain (ICD) as provided herein; and iii) a transmembrane domain linking the ECD to the ICD as provided herein.

[0020] In some embodiments, particles comprising peptides as provided herein are provided. In some embodiments, the particles are viruses or viral vectors. In some embodiments, the viral vector is a lentiviral vector, an AV vector, an AAV vector, etc. In some embodiments, the viral vector is a pseudotyped viral vector.

[0021] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion, and one or more nucleic acid molecules encoding one or more heterologous molecules of interest, wherein the targeting portion comprises a polypeptide having the formula T-S1, wherein T is a target-binding domain and S1 is a stem portion, and wherein the nucleic acid molecule encoding the heterologous molecule of interest is a nucleic acid molecule as provided herein.

[0022] In some embodiments, the stem portion S1 comprises a variant Fc protein. In some embodiments, the variant Fc protein comprises a transmembrane domain, such as, but not limited to, a CD8 or CD28 transmembrane domain. In some embodiments, the variant Fc protein comprises an effector mutation, wherein the effector mutation inhibits the interaction between the Fc protein and Fc-interacting proteins, such as FcγR, C1q, FcRβ, or FcRn.

[0023] In some implementations, the variant Fc protein is a variant IgG1 Fc protein containing one or more mutations selected from the group consisting of: L234A, L235A, N297A, P329G, I253A, H310A, and H435A.

[0024] In some embodiments, the variant IgG1 Fc protein comprises an amino acid sequence having at least 80% identity with SEQ ID NO: 104, at least 85% identity with SEQ ID NO: 104, at least 90% identity with SEQ ID NO: 104, at least 95% identity with SEQ ID NO: 104, at least 98% identity with SEQ ID NO: 104, or at least 100% identity with SEQ ID NO: 104.

[0025] In some implementations, the variant Fc protein is a variant IgG2 Fc protein containing one or more mutations selected from the group consisting of: N297A, P329G, I253A, H310A, and H435A.

[0026] In some implementations, the variant Fc protein is a variant IgG4 Fc protein containing one or more mutations selected from the group consisting of: S228P, L235E, N297A, P329G, I253A, H310A, and H435A.

[0027] In some embodiments, the targeting portion having formula T-S1 includes a stem portion S1 having formula L1-Fc-L2-X1, wherein L1 is a linker or is absent, Fc is a variant Fc protein, L2 is a linker or is absent, and X1 is a polypeptide containing a transmembrane domain, wherein the targeting portion having formula T-S1 has formula T-L1-Fc-L2-X1.

[0028] In some embodiments, the polypeptide containing the transmembrane domain (X1) comprises a peptide having the formula ECD-T M -ICD polypeptides, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M It is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain or a protein that promotes the incorporation of the target portion into the viral particle envelope, or it is absent; wherein the target portion having the formula T-L1-Fc-L2-X1 has the formula T-L1-Fc-L2-ECD-T M -ICD.

[0029] In some embodiments, the stem portion S1 comprises formula L3-X1, wherein L3 is a flexible peptide linker and X1 is a polypeptide containing a transmembrane domain; wherein the targeting portion having formula T-S1 has formula T-L3-X1.

[0030] In some embodiments, the polypeptide containing the transmembrane domain (X1) comprises a peptide having the formula ECD-T M-ICD polypeptides, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M It is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain or a protein that promotes the incorporation of the target portion into the viral particle envelope, or it is absent; wherein the target portion having the formula T-L3-X1 has the formula T-L3-ECD-T M -ICD.

[0031] In some embodiments, the targeting portion binds to CD7. In some embodiments, the targeting portion comprises a polypeptide having a heavy chain variable region comprising HCDR1 of SEQ ID NO: 30, HCDR2 of SEQ ID NO: 31, and HCDR3 of SEQ ID NO: 32, or a variant thereof. In some embodiments, the targeting portion comprises a polypeptide having a light chain variable region comprising LCDR1 of SEQ ID NO: 33, LCDR2 of SEQ ID NO: 34, and LCDR3 of SEQ ID NO: 35, or a variant thereof. In some embodiments, the heavy chain comprises a heavy chain variable region having at least 90% sequence identity with SEQ ID NO: 36. In some embodiments, the light chain comprises a light chain variable region having at least 90% sequence identity with SEQ ID NO: 37. In some embodiments, the CD7-binding targeting portion comprises a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 38. In some embodiments, the target portion that binds to CD7 comprises a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 39.

[0032] In some embodiments, the targeting portion binds to CD8. In some embodiments, the targeting portion comprises a polypeptide having a heavy chain variable region comprising HCDR1 of SEQ ID NO: 42, HCDR2 of SEQ ID NO: 43, and HCDR3 of SEQ ID NO: 44, or a variant thereof. In some embodiments, the targeting portion comprises a polypeptide having a light chain variable region comprising LCDR1 of SEQ ID NO: 45, LCDR2 of SEQ ID NO: 46, and LCDR3 of SEQ ID NO: 47, or a variant thereof. In some embodiments, the heavy chain comprises a heavy chain variable region having at least 90% sequence identity with SEQ ID NO: 48. In some embodiments, the light chain comprises a light chain variable region having at least 90% sequence identity with SEQ ID NO: 49. In some embodiments, the CD8-binding targeting portion comprises a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 50. In some embodiments, the target portion that binds to CD8 comprises a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 51.

[0033] In some implementations, the heteroviral glycoprotein is the SVCV-G polypeptide as provided herein.

[0034] In some embodiments, the heteroviral glycoprotein is the VSV-G polypeptide. In some embodiments, the VSV-G polypeptide comprises substitutions at positions I182, T214, and T352 of SEQ ID NO: 2. In some embodiments, the substitution at position 182 is I182D or I182E. In some embodiments, the substitution at position 214 is T214N. In some embodiments, the substitution at position 352 is T352A.

[0035] In some embodiments, the viral particles provided herein also comprise nucleic acid molecules encoding a heterologous molecule of interest. In some embodiments, the heterologous molecule of interest is as provided herein. In some embodiments, the heterologous molecule of interest is a CAR. In some embodiments, the CAR comprises an antigen-binding domain having a heavy chain variable region having at least 95% identity with SEQ ID NO: 89 and a light chain variable region having at least 95% identity with SEQ ID NO: 90. In some embodiments, the CAR comprises an antigen-binding domain having a heavy chain variable region having at least 95% identity with SEQ ID NO: 94 and a light chain variable region having at least 95% identity with SEQ ID NO: 95.

[0036] In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence having at least 95% identity with SEQ ID NO: 92. In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence having at least 95% identity with SEQ ID NO: 93. In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence having at least 95% identity with SEQ ID NO: 96. In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence having at least 95% identity with SEQ ID NO: 97.

[0037] In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity with SEQ ID NO: 99, at least 90% identity with SEQ ID NO: 99, at least 95% identity with SEQ ID NO: 99, at least 99% identity with SEQ ID NO: 99, or at least 100% identity with SEQ ID NO: 99.

[0038] In some embodiments, a method for infecting cells is provided. In some embodiments, the method includes contacting the cells with viral particles as provided herein.

[0039] In some embodiments, a method for infecting a subject's cells is provided. In some embodiments, the method includes administering to the subject a pharmaceutical composition comprising viral particles as provided herein.

[0040] In some embodiments, methods for delivering a heterologous molecule of interest into cells are provided. In some embodiments, the method includes contacting the cells with viral particles as provided herein, wherein the viral particles contain nucleic acid molecules encoding the heterologous molecule of interest.

[0041] In some embodiments, a method is provided for delivering a heterologous molecule of interest to a subject's cells. In some embodiments, the method includes administering to a subject a viral particle, as provided herein, wherein the viral particle contains a nucleic acid molecule encoding the heterologous molecule of interest.

[0042] In some embodiments, a method for treating a subject's disease or condition is provided. In some embodiments, the method includes administering to the subject a viral particle as provided herein, wherein the viral particle contains a nucleic acid molecule encoding a heterologous molecule of interest for treating the disease or condition.

[0043] In some embodiments, a method for delivering a heterologous molecule to target cells is provided. In some embodiments, the method includes contacting the cells with viral particles as provided herein, wherein the viral particles contain nucleic acid molecules encoding the heterologous molecule.

[0044] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heteroviral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heteroviral molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heteroviral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the target-binding domain comprises a sequence selected from SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stem portion S1 comprises a variant Fc protein comprising a sequence as a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28. In some embodiments, the variant of SEQ ID NO: 26 comprises one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant of SEQ ID NO: 28 comprises one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant Fc protein further comprises a transmembrane domain containing a sequence selected from SEQ ID NO: 61 or SEQ ID NO: 62. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is a nucleic acid molecule as provided herein.

[0045] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heteroviral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heteroviral molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heteroviral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the target-binding domain comprises a sequence selected from SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stem portion S1 comprises the formula L1-Fc-L2-X1, wherein L1 is a linker comprising a sequence selected from SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, or SEQ ID NO: 76, or is absent; Fc is a variant Fc protein comprising a sequence as a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28; L2 is a linker comprising a sequence selected from SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, or SEQ ID NO: 76, or is absent; and X1 is a polypeptide comprising a transmembrane domain. In some embodiments, the variant of SEQ ID NO:26 comprises one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant of SEQ ID NO:27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A. In some embodiments, the variant of SEQ ID NO:28 comprises one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the polypeptide comprising the transmembrane domain (X1) has the formula ECD-T. M-ICD, wherein the ECD is an extracellular domain having a sequence selected from SEQ ID NO:59 or SEQ ID NO:60, or a fragment thereof, or is absent; T M It is a transmembrane domain or fragment thereof having the sequence of SEQ ID NO: 61 or SEQ ID NO: 62; and the ICD is an intracellular domain or a protein that facilitates the incorporation of the target portion into the envelope of the viral particle, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is a nucleic acid molecule as provided herein.

[0046] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heteroviral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heteroviral molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heteroviral glycoprotein comprises a sequence selected from SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the target-binding domain comprises a sequence selected from SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stem portion S1 comprises the formula L1-Fc-L2-X1, wherein L1 is a linker containing the sequence of SEQ ID NO: 55 or is absent; Fc is a variant Fc protein containing a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27 or SEQ ID NO: 28, L2 is a linker containing the sequence of SEQ ID NO: 55 or is absent; and X1 is a polypeptide containing a transmembrane domain. In some embodiments, the variant of SEQ ID NO: 26 comprises one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A and H435A. In some embodiments, the variant of SEQ ID NO: 27 comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A and H435A. In some embodiments, the variant of SEQ ID NO: 28 comprises one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the polypeptide comprising a transmembrane domain (X1) has the formula ECD-T.M -ICD, wherein ECD is an extracellular domain or a fragment thereof having the sequence of SEQ ID NO: 60, or is absent; T M It is a transmembrane domain or fragment thereof having the sequence of SEQ ID NO: 62; and the ICD is an intracellular domain or a protein that facilitates the incorporation of the target portion into the envelope of a viral particle, wherein the ICD contains an env incorporation motif having the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is a nucleic acid molecule as provided herein.

[0047] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heterologous viral glycoprotein comprises a sequence selected from SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stem portion S1 comprises the formula L1-Fc-L2-X1, wherein L1 is a linker comprising the sequence of SEQ ID NO: 55; Fc is a variant Fc protein comprising the sequence of SEQ ID NO: 104; L2 is a linker or is absent; and X1 is a polypeptide comprising a transmembrane domain. In some embodiments, the polypeptide comprising the transmembrane domain (X1) has the formula ECD-T. M -ICD, where ECD is an extracellular domain having the sequence SEQ ID NO: 60; T M It is a transmembrane domain having the sequence of SEQ ID NO: 62; and the ICD is an intracellular domain or a protein that facilitates the incorporation of a targeted portion into the envelope of a viral particle, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest encodes a chimeric antigen receptor and polypeptide as provided herein, the polypeptide comprising i) a ligand-binding extracellular domain (ECD) as provided herein, ii) an immune cell activation intracellular domain (ICD) as provided herein, and ii) a transmembrane domain linking the ECD to the ICD as provided herein.

[0048] In some embodiments, a viral particle is provided, wherein the viral particle comprises a heterologous viral glycoprotein, a targeting portion, and a nucleic acid molecule encoding a heterologous molecule of interest. In some embodiments, the heterologous viral glycoprotein comprises an amino acid sequence having at least 90% identity with SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identity with SEQ ID NO: 23 or SEQ ID NO: 25, at least 99% identity with SEQ ID NO: 23 or SEQ ID NO: 25, or at least 100% identity with SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the targeting portion comprises an amino acid sequence having at least 90% identity with SEQ ID NO: 98, at least 95% identity with SEQ ID NO: 98, at least 99% identity with SEQ ID NO: 98, or at least 100% identity with SEQ ID NO: 98. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest encodes a chimeric antigen receptor and a polypeptide as provided herein, the polypeptide comprising i) a ligand-binding extracellular domain (ECD) as provided herein, ii) an immune cell activation intracellular domain (ICD) as provided herein, and ii) a transmembrane domain linking the ECD to the ICD as provided herein.

[0049] In some embodiments, a viral particle is provided, wherein the viral particle comprises a heterologous viral glycoprotein, a targeting portion, and a nucleic acid molecule encoding a heterologous molecule of interest. In some embodiments, the heterologous viral glycoprotein comprises an amino acid sequence having at least 90% identity with SEQ ID NO: 52 or SEQ ID NO: 53, at least 95% identity with SEQ ID NO: 53, at least 99% identity with SEQ ID NO: 53, or at least 100% identity with SEQ ID NO: 52 or SEQ ID NO: 53. In some embodiments, the targeting portion comprises an amino acid sequence having at least 90% identity with SEQ ID NO: 98, at least 95% identity with SEQ ID NO: 98, at least 99% identity with SEQ ID NO: 98, or at least 100% identity with SEQ ID NO: 98. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest encodes a chimeric antigen receptor and a polypeptide as provided herein, the polypeptide comprising i) a ligand-binding extracellular domain (ECD) as provided herein, ii) an immune cell activation intracellular domain (ICD) as provided herein, and ii) a transmembrane domain linking the ECD to the ICD as provided herein.

[0050] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO:38 or SEQ ID NO:39. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible peptide linker comprising the amino acid sequence of SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, or SEQ ID NO:58, and X1 is a peptide linker comprising a transmembrane domain. In some embodiments, the polypeptide (X1) comprising the transmembrane domain has the formula ECD-T. M -ICD, wherein ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent, T M The ICD is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and is an intracellular domain or a protein that facilitates the incorporation of the target portion into the viral particle envelope, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is the nucleic acid molecule as provided herein.

[0051] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO:50 or SEQ ID NO:51. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible peptide linker comprising the amino acid sequence of SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, or SEQ ID NO:58, and X1 is a peptide linker comprising a transmembrane domain. In some embodiments, the polypeptide (X1) comprising the transmembrane domain has the formula ECD-T. M-ICD, wherein ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent, T M The ICD is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and is an intracellular domain or a protein that facilitates the incorporation of the target portion into the viral particle envelope, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is the nucleic acid molecule as provided herein.

[0052] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heterologous viral glycoprotein comprises the sequence of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible peptide linker comprising the amino acid sequence of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, or SEQ ID NO: 58, and X1 is a peptide linker comprising a transmembrane domain. In some embodiments, the polypeptide (X1) containing a transmembrane domain has the formula ECD-T M -ICD, where ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent, T M The ICD is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and is an intracellular domain or a protein that facilitates the incorporation of the target portion into the viral particle envelope, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is the nucleic acid molecule as provided herein.

[0053] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heterologous viral glycoprotein comprises the sequence of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 52, or SEQ ID NO: 53. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 50, or SEQ ID NO: 51. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible peptide linker comprising the amino acid sequence of SEQ ID NO: 55, and X1 is a peptide linker comprising a transmembrane domain. In some embodiments, the polypeptide (X1) comprising the transmembrane domain has the formula ECD-T. M -ICD, wherein ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, or a fragment thereof, or is absent, T M The ICD is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and is an intracellular domain or a protein that facilitates the incorporation of the target portion into the viral particle envelope, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or the ICD is absent. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is the nucleic acid molecule as provided herein.

[0054] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heterologous viral glycoprotein comprises the sequence of SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible peptide linker comprising the amino acid sequence of SEQ ID NO: 55, and X1 is a polypeptide linker comprising a transmembrane domain. In some embodiments, the polypeptide (X1) comprising the transmembrane domain has the formula ECD-T. M-ICD, where ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59, T M It is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61, and the ICD is an intracellular domain or a protein that facilitates the incorporation of a target portion into the envelope of a viral particle, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is the nucleic acid molecule as provided herein.

[0055] In some embodiments, a viral particle is provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting portion comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein T is a target-binding domain and S1 is a stem portion. In some embodiments, the heterologous viral glycoprotein comprises the sequence of SEQ ID NO: 52 or SEQ ID NO: 53. In some embodiments, the target-binding domain comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the stem portion S1 comprises the formula L3-X1, wherein L3 is a flexible peptide linker comprising the amino acid sequence of SEQ ID NO: 55, and X1 is a peptide linker comprising a transmembrane domain. In some embodiments, the polypeptide (X1) comprising the transmembrane domain has the formula ECD-T. M -ICD, where ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59, T M The ICD is a transmembrane domain containing the amino acid sequence of SEQ ID NO: 61, and is an intracellular domain or a protein that facilitates the incorporation of a target portion into the envelope of a viral particle, wherein the ICD contains an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest encodes a chimeric antigen receptor and polypeptide as provided herein, the polypeptide comprising i) a ligand-binding extracellular domain (ECD) as provided herein, ii) an immune cell activation intracellular domain (ICD) as provided herein, and ii) a transmembrane domain linking the ECD to the ICD as provided herein.

[0056] In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity with SEQ ID NO: 99, at least 90% identity with SEQ ID NO: 99, at least 95% identity with SEQ ID NO: 99, at least 99% identity with SEQ ID NO: 99, or at least 100% identity with SEQ ID NO: 99.

[0057] This application also relates to the following implementation schemes numbered below.

[0058] 1. A particle or composition comprising one or more nucleic acid molecules encoding one of the following:

[0059] a) Chimeric antigen receptor (CAR); and

[0060] b) A polypeptide, wherein the polypeptide comprises:

[0061] i) Ligand-binding extracellular domain (ECD);

[0062] ii) Immune cell activation intracellular domain (ICD); and

[0063] iii) A transmembrane structural domain connecting the ECD and the ICD.

[0064] 2. The particle according to embodiment 1, wherein the immune cell activating ICD comprises an ICD of hGCSF-R or a mutant or fragment thereof.

[0065] 3. The particle according to embodiment 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111, or an amino acid sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

[0066] 4. The particle according to embodiment 3, wherein the hGCSF-R ICD does not contain the sequence of SEQ ID NO:110.

[0067] 5. The particle according to embodiment 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111 and does not contain amino acid residues 800-813, 775-813, 750-813, 725-813 or 716-813 of SEQ ID NO: 125 or fragments thereof located at the C-terminal residue of SEQ ID NO: 111.

[0068] 6. The particle according to embodiment 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111 and amino acid residues 716-812 of SEQ ID NO: 125 or a fragment thereof located at the C-terminal residue of SEQ ID NO: 111, provided that the ICD does not contain a continuous fragment of amino acid residues 716-813 of SEQ ID NO: 125.

[0069] 7. The particle according to embodiment 1, wherein the ECD is a GCSF-R ECD, optionally wherein the GCSF-R ECD is a human GCSF-R ECD containing the amino acid sequence of SEQ ID NO: 107 or SEQ ID NO: 108.

[0070] 8. The particle according to embodiment 1, wherein the ECD comprises an ECD of human growth hormone receptor (hGH-R) or a mutant or fragment thereof, wherein the ECD of human growth hormone receptor (hGH-R) comprises an amino acid sequence encoded by exons 1-7 of hGH-R or a mutant or fragment thereof.

[0071] 9. The particle according to embodiment 8, wherein the ECD of the hGH-R contains a mutation of C259R.

[0072] 10. The particles according to embodiment 8, wherein the ECD of the hGH-R comprises the amino acid sequence of SEQ ID NO: 114 or SEQ ID NO: 115, or an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

[0073] 11. The particle according to embodiment 1, wherein the ICD comprises an ICD of glycoprotein 130 (“gp130ICD”) or a variant or fragment thereof.

[0074] 12. The particle according to embodiment 1, wherein the ICD comprises a fragment of the GCSF-R ICD, wherein the GCSF-R ICD fragment does not contain the STAT3, SH3 and / or SHC structural domains.

[0075] 13. The particle according to embodiment 1, wherein the ICD comprises IL-2RβICD or a fragment or variant thereof.

[0076] 14. The particles according to embodiment 1, wherein the ICD comprises IL-7RαICD or a fragment or variant thereof.

[0077] 15. The particle according to embodiment 1, wherein the ICD comprises a gp130 ICD or a variant or fragment thereof linked to an IL-2Rβ ICD or a fragment thereof.

[0078] 16. The particle according to embodiment 1, wherein the transmembrane domain comprises a GCSF-R, gp130, or hGH-R transmembrane domain.

[0079] 17. The particle according to embodiment 1, wherein the polypeptide comprises:

[0080] a) ECDs as GCSFR-ECDs or variants thereof; transmembrane domains as GCSF-R transmembrane domains or variants thereof; and immune-activated ICDs as GCSF-R ICDs or variants thereof;

[0081] b) ECDs as hGH-R-ECDs or variants thereof; transmembrane domains as GCSF-R transmembrane domains or variants thereof; and immune-activated ICDs as GCSF-R ICDs or variants thereof;

[0082] c) ECDs as hGH-R-ECDs or variants thereof; transmembrane domains as gp130 transmembrane domains or variants thereof; and immune-activated ICDs containing gp130 ICDs or variants thereof and IL-2Rβ ICDs or variants thereof; or

[0083] d) ECDs as hGH-R-ECD or variants thereof; transmembrane domains as GCSF-R transmembrane domains or variants thereof; and immune-activated ICDs containing GCSF-R ICDs or variants thereof and IL-7Rα ICDs or variants thereof.

[0084] 18. The particle according to embodiment 1, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120 or SEQ ID NO: 124, or a variant thereof.

[0085] 19. The particles according to embodiment 18, wherein the variant comprises a polypeptide having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124.

[0086] 20. The particle according to embodiment 1, wherein the CAR comprises an antigen-binding domain that binds to CD20.

[0087] 21. The particle according to embodiment 20 of the numbered scheme, wherein the CAR comprises an amino acid sequence having at least 85% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, at least 90% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, at least 95% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, or at least 99% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, or at least 100% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129.

[0088] 22. The particle according to any one of embodiments 1 to 21, wherein the particle is a vector, virus, liposome, plasmid or cell.

[0089] 23. The particle according to any one of embodiments 1 to 22, wherein the particle is a viral vector, and wherein the viral vector is a lentiviral vector, an AV vector, an AAV vector, etc.

[0090] 24. A nucleic acid molecule encoding a polypeptide, said nucleic acid molecule comprising isolated nucleic acid molecules, said polypeptide comprising:

[0091] i) Ligand-binding extracellular domain (ECD);

[0092] ii) Immune cell activation intracellular domain (ICD); and

[0093] iii) A transmembrane structural domain connecting the ECD and the ICD.

[0094] 25. The nucleic acid molecule according to embodiment 24, wherein the nucleic acid molecule encodes a polypeptide according to any one of embodiments 1 to 19.

[0095] 26. The nucleic acid molecule according to embodiment 24, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120 or SEQ ID NO: 124, or a variant thereof.

[0096] 27. The nucleic acid molecule according to embodiment 26, wherein the variant comprises a polypeptide having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 16, or SEQ ID NO: 20.

[0097] 28. The nucleic acid molecule according to any one of embodiments 24 to 27, wherein the nucleic acid molecule further encodes a chimeric antigen receptor (CAR).

[0098] 29. A viral particle comprising a heterologous viral glycoprotein, a targeting portion, and one or more nucleic acid molecules encoding one or more heterologous molecules of interest.

[0099] The targeting portion comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stem portion.

[0100] The nucleic acid molecule encoding the heterologous molecule of interest is a nucleic acid molecule according to any one of the numbered embodiments 24 to 28.

[0101] 30. The viral particle according to embodiment 29, wherein the target-binding domain (T) binds to: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR α, TCR β, TCR γ, TCR δ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7 or CXCR3; glycosylated CD43 epitopes expressed in acute leukemia or lymphoma but not on hematopoietic progenitor cells; glycosylated CD43 epitopes expressed in non-hematopoietic cancers; A kinase anchoring protein 4 (AKAP-4); adrenergic receptor β3 (ADRB3); AFP; anaplastic lymphoma kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie2); anti-desmosome core protein 1 (Dsg1) autoantibody; anti-desmosome core protein 3 (Dsg3) autoantibody; B7H3 (CD276); biotin; bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; cancer / testis antigen 1 (NY-ESO) -1); Cancer / Testis Antigen 2 (LAGE-1a); Carbonic Anhydrase IX (CA1X); Carcinoembryonic Antigen (CEA); CCCTC Binding Factor (Zinc Finger Protein) Like (BORIS or Imprinted Site Regulation Sibling Factor); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32 (FCGR2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGL11; CD200R; CD276 / B7H3; CD300 Molecular-like Family Member f (CD300LF); CDH1-CD324; CDH6; CDH17; CDH19; Chromosome X Open Reading Frame 61 (CXORF61); Seal Protein 6 (CLDN6); Seal Protein 1 8.2 (CLD18A2 or CLDN18A.2); CMV pp65; C-MYC epitope tag; Cripto; CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24); CSF2RA (GM-CSFR-α); C-type lectin domain family 12 member A (CLEC12A); C-type lectin-like molecule-1 (CLL-1 or CLECL1); Cyclin B1; Cytochrome P450 IB 1 (CYP1B 1); DLL3; EBV-EBNA3c; EGF-bke module 2 containing mucin-like hormone receptor-like receptor (EMR2); Elongation factor 2 mutant (ELF2M); Hepatin B2;Hepatic glycoprotein A receptor 2 (EphA2); Epidermal growth factor receptor (EGFR); Epidermal growth factor receptor variant III (EGFRviii); Epithelial cell adhesion molecule (EPCAM); ERG; ETS translocation variant gene 6 located on chromosome 12p (ETV6-AML); Fc fragment of IgA receptor (FCAR or CD89); Fc receptor-like 5 (FCRL5); Fibroblast activation protein α (FAP); FITC; Fms-like tyrosine kinase 3 (FLT3); Folate receptor α (Fra or FR 1); Follicle receptor β (FRb); Follicle-stimulating hormone receptor (FSHR); Fos-associated antigen 1; Fucosyl-GM1; G protein-coupled receptor class C5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; Ganglioside G2 (GD2); Ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(l-4)bDGlcp(ll)Cer); Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l- 4) bDGlcp(ll)Cer); GD3; GFRα4; Glycoprotein 100 (gplOO); Phosphatidylinositol proteoglycan-3 (GPC3); Gonadotropin receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylate cyclase C (GCC); Mutant heat shock protein 70-2 (muthsp70-2); Hepatitis A virus cell receptor 1 (HAVCR1); GloboH glycosylceramide hexasaccharide moiety (GloboH); High molecular weight melanoma-associated antigen (HMWMAA); HIV1 envelope glycoprotein; HLA; HLA-DOA; HLA-A; HLA-A2; HLA-B; HLA-C; HLA-DM; HLA-DOB; HLA-DP; HLA-DQ; HLA-DR; HLA-G; HTLV1-Tax; Human papillomavirus E6 (HPVE6); Human papillomavirus E7 (HPV E7); human telomerase reverse transcriptase (hTERT); IgE; IL13Ra2; IL11Ra; immunoglobulin λ-like polypeptide 1 (IGLL1); influenza A hemagglutinin (HA); insulin-like growth factor 1 receptor (IGF-I receptor); interleukin 11 receptor α (IL-11Ra); interleukin 13 receptor subunit α-2 (IL-13Ra2 or CD213A2); intestinal carboxylesterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; pod protein; leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); luteinizing hormone receptor (LHR); Lewis (Y) antigen; Lews Ag; Livl; locus K9 (LY6K)Low-conductivity chloride channels; Lymphocyte antigen 6 complex; Lymphocyte antigen 75 (LY75); Lymphocyte-specific protein tyrosine kinase (LCK); Breast differentiation antigen (NY-BR-1); T-cell recognized melanoma antigen 1 (MelanA or MARTI); Melanoma-associated antigen 1 (MAGE-A1); Melanoma cancer testis antigen-1 (MAD-CT-1); Melanoma cancer testis antigen-2 (MAD-CT-2); Melanoma cell apoptosis inhibitor (ML-IAP); Mesothelin; MPL; Cell surface-associated mucin 1 (MUC1); N-acetylglucosamine transferase V (NA17); Ligin-4; Neural cell adhesion molecule (NCAM); NKG2D; N YBR1; O-acetyl-GD2 ganglioside (OacGD2); olfactory receptor 51E2 (OR51E2); oncogene fusion protein composed of breakpoint cluster region (BCR) and Abelson murine leukemia virus oncogene homolog 1 (Abl) (bcr-abl); P53 mutant; pairing box protein Pax-3 (PAX3); pairing box protein Pax-5 (PAX5); pan-connector protein 3 (PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor β (PDGFR-β); polysialic acid; proapocrine-binding protein sp32 (OY-TES1); prostate enzyme; prostate cancer tumor antigen-1 (PCT) A-1 or galactohemagglutinin 8); prostate stem cell antigen (PSCA); prostate-specific membrane antigen (PSMA); prostatic acid phosphatase (PAP); Prostein; serine protease 21 (Testisin or PRSS21); proteasome macropain subunit β9 (LMP2); PTK7; Ras G12V; Ras homolog family member C (RhoC); Rat sarcoma (Ras) mutant; Receptor to advanced glycation end products (RAGE-1); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Receptor tyrosine protein kinase ERBB2 or Her-22 / neu; Renal pervasive protein 1 (RU1); Renal pervasive protein 2 (RU2); Sarcoma translocation breakpoint; Serine 2 (TMPRSS2) ETS fusion gene; Sialoyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialic acid Lewis antigen); Sperm protein 17 (SPA17); T cell recognized squamous cell carcinoma antigen 3 (SART3); Stage-specific embryonic antigen-4 (SSEA-4); STEAP1; Survival protein; Synovial sarcoma X breakpoint 2 (SSX2); TCR γ-substituted reading frame protein (TARP); TCR-β1 chain; TCR-β2 chain; TCR-δ chain; TCR-γ chain; TCRγ-δ; telomerase; TGFβR2;Antigens recognized by TNT antibodies: thyroid-stimulating hormone receptor (TSHR); Timl- / HVCR1; tissue factor 1 (TF1); Tn ag; Tn antigen ((Tn Ag) or (GalNAca-Ser / Thr)); TNF receptor family member B cell maturation (BCMA); transglutaminase 5 (TGS5); transmembrane protease; TROP2; tumor endothelial marker 1 (TEM1 / CD248); tumor endothelial marker 7-associated (TEM7R); tumor protein p53 (p53); tumor-associated glycoprotein 72 (TAG72); tyrosinase; tyrosinase-associated protein 2 (TRP-2); urolytic protein 2 (UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein (WT1); or X antigen family member 1A (XAGE1).

[0102] 31. The viral particle according to embodiment 30, wherein the target-binding domain (T) binds to CD7.

[0103] 32. The viral particle according to embodiment 31, wherein the target-binding domain (T) binding to CD7 comprises a polypeptide, the polypeptide comprising a sequence having at least 90% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, at least 95% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, at least 99% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, or a sequence as shown in SEQ ID NO: 38 or SEQ ID NO: 39.

[0104] 33. The viral particle according to any one of embodiments 29 to 32, wherein the heterologous viral glycoprotein is a VSV-G polypeptide.

[0105] 34. The viral particle according to embodiment 33, wherein the VSV-G polypeptide comprises the sequence of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24 or SEQ ID NO:25.

[0106] 35. A viral particle comprising a heterologous viral glycoprotein, a targeting portion, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein;

[0107] The heteroviral glycoprotein comprises an amino acid sequence having at least 90% identity with SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identity with SEQ ID NO: 23 or SEQ ID NO: 25, at least 99% identity with SEQ ID NO: 23 or SEQ ID NO: 25, or at least 100% identity with SEQ ID NO: 23 or SEQ ID NO: 25.

[0108] The targeting portion comprises an amino acid sequence having at least 90% identity with SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, at least 95% identity with SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, at least 99% identity with SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, or at least 100% identity with SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127; and

[0109] The nucleic acid molecule that encodes the heterologous molecule of interest

[0110] Encoding a chimeric antigen receptor and a polypeptide, said polypeptide comprising:

[0111] i) Ligand-binding extracellular domain (ECD);

[0112] ii) Immune cell activation intracellular domain (ICD); and

[0113] iii) A transmembrane structural domain connecting the ECD and the ICD.

[0114] 36. The viral particle according to embodiment 35, wherein the CAR comprises an amino acid sequence having at least 85% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, at least 90% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, at least 95% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, at least 99% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129, or at least 100% identity with SEQ ID NO: 99, SEQ ID NO: 128 or SEQ ID NO: 129.

[0115] 37. The viral particle according to embodiment 35 or 36, wherein the polypeptide encoded by the nucleic acid molecule comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120 or SEQ ID NO: 124, or a variant thereof.

[0116] 38. The viral particle according to any one of embodiments 35 to 37, wherein the chimeric antigen receptor and the polypeptide are encoded by separate nucleic acid molecules.

[0117] 39. The viral particle according to any one of embodiments 35 to 37, wherein the chimeric antigen receptor and the polypeptide are encoded by the same nucleic acid molecule.

[0118] 40. The viral particle according to embodiment 39, wherein the chimeric antigen receptor and the polypeptide are separated by a sequence encoding a cleavable peptide such as peptide 2A.

[0119] 41. A method of infecting, transfecting, or transducing cells, the method comprising contacting the cells with particles according to any one of embodiments 1 to 23 or viral particles according to any one of embodiments 29 to 40.

[0120] 42. A method of delivering one or more heterologous molecules of interest into a cell, the method comprising contacting the cell with a particle according to any one of embodiments 1 to 23 or a viral particle according to any one of embodiments 29 to 40, wherein the viral particle contains a nucleic acid molecule encoding the heterologous molecule of interest.

[0121] 43. The method according to embodiment 41 or 42, wherein the method further comprises administering to the cells or to a subject one or more compounds, such as, but not limited to, growth hormone, G-CSF, granocyte, filgrastim (Neupogen, Zarcio, Nivestim, Accofil), long-acting (pegylated) filgrastim (Neulasta, Pelmeg, Ziextenco), or risperidone (Lonquex).

[0122] 44. A method of treating a disease or condition of a subject, the method comprising administering to the subject a particle according to any one of embodiments 1 to 23 or a viral particle according to any one of embodiments 29 to 40, wherein the viral particle contains a nucleic acid molecule encoding a heterologous molecule of interest for treating the disease or condition, wherein the nucleic acid molecule encoding the heterologous molecule of interest is a nucleic acid molecule according to any one of embodiments 24 to 28.

[0123] 45. A method for treating a disease or ailment of a subject, the method comprising:

[0124] i) Administering to the subject the particles according to any one of embodiments 1 to 23 or the viral particles according to any one of embodiments 29 to 40, and

[0125] ii) Administering an effective amount of one or more compounds capable of binding to the ECD of human GCSFR to the subject, thereby treating the disease or condition.

[0126] The viral particles contain one or more nucleic acid molecules encoding one or more heterologous molecules of interest for the treatment of the disease or condition.

[0127] The one or more nucleic acid molecules encoding one or more heterologous molecules of interest are one or more nucleic acid molecules according to any one of the numbered embodiments 24 to 28.

[0128] 46. ​​A method for treating a disease or ailment of a subject, the method comprising:

[0129] i) Administering to the subject the particles according to any one of embodiments 1 to 23 or the viral particles according to any one of embodiments 29 to 40, and

[0130] ii) Administering an effective amount of one or more compounds capable of binding to the ECD of human GHR to the subject, thereby treating the disease or condition.

[0131] The viral particles contain one or more nucleic acid molecules encoding one or more heterologous molecules of interest for the treatment of the disease or condition.

[0132] The one or more nucleic acid molecules encoding one or more heterologous molecules of interest are one or more nucleic acid molecules according to any one of the numbered embodiments 24 to 28.

[0133] 47. A method for treating a disease or ailment of a subject, the method comprising:

[0134] i) Administering the subject with the viral particles according to any one of embodiments 29 to 40, and

[0135] ii) Administering an effective amount of one or more compounds capable of binding to the ECD of human GHR to the subject, thereby treating the disease or condition.

[0136] The viral particles contain nucleic acid molecules encoding a heterologous molecule of interest for the treatment of the disease or condition.

[0137] The nucleic acid molecule or plurality of nucleic acid molecules encoding the heterologous molecule of interest encodes a chimeric antigen receptor and a polypeptide, wherein the polypeptide comprises:

[0138] i) Ligand-binding extracellular domain (ECD);

[0139] ii) Immune cell activation intracellular domain (ICD); and

[0140] iii) A transmembrane structural domain connecting the ECD and the ICD.

[0141] 48. The method according to embodiment 47 of the numbered scheme, wherein the CAR comprises an amino acid sequence having at least 85% identity with SEQ ID NO: 99, at least 90% identity with SEQ ID NO: 99, at least 95% identity with SEQ ID NO: 99, at least 99% identity with SEQ ID NO: 99, or at least 100% identity with SEQ ID NO: 99.

[0142] 49. The method according to embodiment 47 or 48, wherein the polypeptide encoded by the nucleic acid molecule comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120 or SEQ ID NO: 124, or a variant thereof.

[0143] 50. A method of delivering one or more heterologous molecules to a target cell, the method comprising contacting the cell with a particle according to any one of embodiments 1 to 23 or a viral vector or particle according to any one of embodiments 29 to 40, wherein the particle comprises one or more nucleic acid molecules encoding the one or more heterologous molecules. Attached Figure Description

[0144] Figure 1A and Figure 1B An exemplary chimeric GCSFR construct provided in this article is illustrated.

[0145] Figure 2A and Figure 2B The crystal structure of VSV-G combined with LDL-R is illustrated. Figure 2A The crystal structure of VSV-G bound to CR3 of LDL-R is illustrated. Figure 2B The crystal structure of VSV-G with CR2 bonded to LDL-R is illustrated.

[0146] Figure 3A and Figure 3B The effect of adding negatively charged amino acids to the VSV-G:LDL-R binding interface on native affinity and cohesion is illustrated. Figure 3A The titration of the VSV-G construct on SupT1 cells is illustrated. Figure 3B Examples are given by Figure 3A The functional titer of each construct is calculated by titration.

[0147] Figure 4 The comparison of extracellular domains of different VSV-G proteins from different strains is illustrated.

[0148] Figure 5 The effects of various VSV-G mutations on the serum stability of viral constructs combined with CD7 binding compounds are illustrated.

[0149] Figure 6 The effects of various VSV-G mutations on the serum stability of viral constructs combined with CD7 binding compounds are illustrated.

[0150] Figure 7The ability of various rhabdoviral G proteins, alone or in combination with CD7 conjugates, to transduce SupT1 and PBMC cells is illustrated.

[0151] Figures 8A-8L Flow cytometry data of human PBMCs transduced with an exemplary vector containing a CD7 conjugate as disclosed herein are shown.

[0152] Figure 8M Flow cytometry data of human PBMCs transduced with an exemplary vector containing a CD7 conjugate as disclosed herein are shown.

[0153] Figure 9A -L shows flow cytometry data of non-human primate PBMCs transduced with an exemplary vector containing CD7 conjugates as disclosed herein.

[0154] Figure 9M Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing CD7 conjugates as disclosed herein are shown.

[0155] Figure 10A Flow cytometry data of human PBMCs transduced with an exemplary vector containing a CD8 conjugate as disclosed herein are shown.

[0156] Figure 10B Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing CD8 conjugates as disclosed herein are shown.

[0157] Figure 11A VSV-G carrying CD7 conjugates with variant Fc stems is illustrated. The ability of pseudotyped lentiviral particles to transduce SupT1 cells and human and non-human primate PBMCs. Figure 11B The transduction of cells in the absence of CD7 binders is illustrated. Figure 11C Transduction of human and non-human primate PBMCs based on MOI calculated by SupT1 titration is illustrated. VSV-G This indicates VSV-G (I182E, T214N, T352A).

[0158] Figure 12A An example is VSV-G carrying a CD7 conjugate with a flexible stem. The ability of pseudotyped lentiviral particles to transduce SupT1 cells and human and non-human primate PBMCs. Figure 12B The transduction of cells in the absence of CD7 binders is illustrated. Figure 12C Transduction of human and non-human primate PBMCs based on MOI calculated by SupT1 titration is illustrated. VSV-G This indicates VSV-G (I182E, T214N, T352A).

[0159] Figure 13A This demonstrates the ability of CD7 conjugates carrying flexible stems of varying lengths to transduce SupT1 cells, compared to other IgG-based conjugates. Figure 13B This demonstrates the ability of CD7 conjugates carrying flexible stems of varying lengths to transduce activated PBMCs compared to other IgG-based conjugates.

[0160] Figure 14A An example is SVCV-G carrying a CD7 conjugate with a variant Fc stem. The ability of pseudotyped lentiviral particles to transduce SupT1 cells and human and non-human primate PBMCs. Figure 14B The transduction of cells in the absence of CD7 binders is illustrated. Figure 14C Transduction of human and non-human primate PBMCs based on MOI calculated by SupT1 titration is illustrated.

[0161] Figure 15A The ability of lentiviral particles carrying CD7 conjugates with flexible stems, pseudotyped SVCV-G, to transduce SupT1 cells and human and non-human primate PBMCs is illustrated. Figure 15B The transduction of cells in the absence of CD7 binders is illustrated. Figure 15C Transduction of human and non-human primate PBMCs based on MOI calculated by SupT1 titration is illustrated.

[0162] Figures 16A-16D It's VSV-G Comparison of the transduction capabilities of pseudotyped lentiviral particles (Figures A and B) and SVCV-G pseudotyped lentiviral particles (Figures C and D) into SupT1 cells and human and non-human primate PBMCs. VSV-G This indicates VSV-G (I182E, T214N, T352A).

[0163] Figures 17A-17D It's VSV-G Comparison of the transduction capabilities of pseudotyped lentiviral particles (Figures A and B) and SVCV-G pseudotyped lentiviral particles (Figures C and D) into SupT1 cells and human and non-human primate PBMCs. VSV-G This indicates VSV-G (I182E, T214N, T352A).

[0164] Figure 18A and Figure 18BThis is a comparison of off-target transduction of GFP in a group of B cell lines compared to the control SupT1 cells. Figure 18A An example is VSV-G carrying a CD7 conjugate with a mutant Fc stem. Data on pseudomorphic lentivirus particles. VSV-G This indicates VSV-G (I182E, T214N, T352A). Figure 18B Data are illustrated for lentiviral particles pseudotyped with SVCV-G carrying CD7 conjugates with mutant Fc stems.

[0165] Figure 19A and Figure 19B This is a comparison of off-target transduction of the CAR20-T2A-GFP construct in a group of B cell lines compared to the control SupT1 cells. Figure 19A An example is VSV-G carrying a CD7 conjugate with a mutant Fc stem. Data on pseudomorphic lentivirus particles. VSV-G This indicates VSV-G (I182E, T214N, T352A). Figure 19B Data are illustrated for lentiviral particles pseudotyped with SVCV-G carrying CD7 conjugates with mutant Fc stems.

[0166] Figure 20A and Figure 20B This is a comparison of off-target transduction of GFP in a group of B cell lines compared to the control SupT1. Figure 20A An example is VSV-G carrying a CD7 conjugate with a flexible stem. Data on pseudomorphic lentivirus particles. VSV-G This indicates VSV-G (I182E, T214N, T352A). Figure 20B Data on lentiviral particles carrying CD7 conjugates with flexible stems and pseudotyped SVCV-G are illustrated.

[0167] Figure 21A and Figure 21B This is a comparison of off-target transduction of the CAR20-T2A-GFP construct in a set of B cell lines compared to the control SupT1. Figure 21A An example is VSV-G carrying a CD7 conjugate with a flexible stem. Data on pseudomorphic lentivirus particles. VSV-G This indicates VSV-G (I182E, T214N, T352A). Figure 21B Data on lentiviral particles carrying CD7 conjugates with flexible stems and pseudotyped SVCV-G are illustrated.

[0168] Figure 22A and Figure 22B Examples of VSV-G transgenics utilizing CD7 conjugates and CD20-CARs as provided herein are illustrated. Pseudotyped lentiviral particles kill Daudi lymphoma cells ( Figure 22A ) or Raji lymphoma cells ( Figure 22B The CAR20 construct utilizes an antigen-binding domain comprising rituximab or, as provided herein, CD20AB1 SEQ ID NO 92 or 93.

[0169] Figures 23A-23F The toxicity of the ligand GCSF to PBMCs transduced using the CAR20-2A-chimeric GCSFR construct as provided herein is illustrated. Figure 23A The results are illustrated in the control group without chimeric receptors. Figure 23B The results for GCSFR-d15 are shown. Figure 23C The results for G7R1 are shown. Figure 23D The results for G21 / R1 are shown. Figure 23E The results for G2R2 are shown. Figure 23F The results for G27 / 2R1 are shown.

[0170] Figures 24A-24E The toxicity of the ligand hGH to PBMCs transduced using the CAR20-2A-chimeric GHR construct as provided herein is illustrated. Figure 24A The results for GHR-GCSFRd715 are shown. Figure 24B The results for GHR-G7R1 are shown. Figure 24C The results for GHR-G21 / 7R1 are shown. Figure 24D The results of GHR-G2R2 are shown. Figure 24E The results for GHR-G27 / 2R1 are shown.

[0171] Figures 25A-25F This illustrates the ability of chimeric GCSFR receptor-induced PBMC proliferation provided in this paper. Figure 25A The results are illustrated in the control group without chimeric receptors. Figure 25B The results for GCSFR-d15 are shown. Figure 25C The results for G7R1 are shown. Figure 25D The results for G21 / R1 are shown. Figure 25E The results for G2R2 are shown. Figure 25F The results for G27 / 2R1 are shown.

[0172] Figures 26A-26E This illustrates the ability of chimeric GHR receptor-induced PBMC proliferation provided in this paper. Figure 26AThe results for GHR-GCSFRd715 are shown. Figure 26B The results for GHR-G7R1 are shown. Figure 26C The results for GHR-G21 / 7R1 are shown. Figure 26D The results of GHR-G2R2 are shown. Figure 26E The results for GHR-G27 / 2R1 are shown.

[0173] Figures 27A-27E The results of subsequent PBMC-induced proliferation experiments using selected chimeric receptor constructs are illustrated. Figure 27A The results for GCSFR FL are shown. Figure 27B The results for GCSFRd715 are shown. Figure 27C The results for GCSFR-G2R2 are shown as an example. Figure 27D The results of GHR-GCSFR are shown as an example. Figure 27E The results for GHR-dFN-GCSFR are shown as an example. Figure 27F The results of GHR-G2R2 are shown.

[0174] Figure 28A and Figure 28B Examples of VSV-G transgenics utilizing CD7 conjugates and CD20-CARs as provided herein are illustrated. The ability of pseudotyped lentiviral particles to deplete B cells in huCD34 NSG mice in vivo. This was assessed by evaluating CD20 (… Figure 28A ) or CD19 ( Figure 28B (To detect B cells)

[0175] Figure 29A and Figure 29B Examples of VSV-G transgenics utilizing CD7 conjugates and CD20-CARs as provided herein are illustrated. The ability of pseudomorphic lentiviral particles to prevent tumor formation in the body. Figure 29A The experimental design was described. Figure 29B Mice that received intravenous lentiviral particles as presented herein prior to Raji tumor infusion had significantly lower tumor burden than mice receiving the control (GFP) vector or untreated mice. Tumor burden was measured via IVIS imaging.

[0176] Figure 30A and Figure 30B Examples of VSV-G transgenics utilizing CD7 conjugates and CD20-CARs as provided herein are illustrated. The ability of pseudomorphic lentiviral particles to eliminate established Raji tumors in vivo. Figure 30A The experimental design was described. Figure 30BThis study illustrates how tumor burden in mice receiving intravenous lentiviral particles as described herein decreased to below the detection limit 6 days after Raji tumor infusion. Tumor burden was measured via IVIS imaging.

[0177] Figure 31A An example CAR-GCSF receptor construct for tunable expansion in cells is shown. Figure 31B Flow cytometry data demonstrating tunable CAR cell proliferation in vitro are presented. Figure 31C The dose-response of CAR-T cells to exogenous GCSF agonists is shown. Figure 31D and Figure 31E An example in vivo experimental design for evaluating the CAR-GSCFR construct is described. Figure 31F and Figure 31G The study demonstrated significant proliferation of CAR20+ cells in the presence of GCSF agonists.

[0178] Figures 32A-32F The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GCSFR construct, or GFP when treated with a specified concentration of GCSF or complete medium are shown. Figure 32A The growth of PBMCs from PBMC donor 1 transduced with a single CAR20 is illustrated. Figure 32B The growth of PBMCs from PBMC donor 2 transduced with a single CAR20 is illustrated. Figure 32C The growth of PBMCs from PBMC donor 1, transduced using CAR20 and GCSFR constructs, is illustrated. Figure 32D The growth of PBMCs from PBMC donor 2 transduced with CAR20 and GCSFR constructs is illustrated. Figure 32E The growth of PBMCs from PBMC donor 1 transduced with GFP and GCSFR constructs is illustrated. Figure 32F The growth of PBMCs from PBMC donor 2 transduced with GFP and GCSFR constructs is illustrated.

[0179] Figures 33A-33F The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GHR construct, or GFP when treated with a specified concentration of GH or complete medium are shown. Figure 33A The growth of PBMCs from PBMC donor 1 transduced with a single CAR20 is illustrated. Figure 33B The growth of PBMCs from PBMC donor 2 transduced with a single CAR20 is illustrated. Figure 33C The growth of PBMCs from PBMC donor 1 transduced with CAR20 and GHR constructs is illustrated. Figure 33DThe growth of PBMCs from PBMC donor 2 transduced with CAR20 and GHR constructs is illustrated. Figure 33E The growth of PBMCs from PBMC donor 1 transduced with GFP and GHR constructs is illustrated. Figure 33F The growth of PBMCs from PBMC donor 2 transduced with GFP and GHR constructs is illustrated. Detailed Implementation

[0180] This document provides particles or compositions comprising one or more nucleic acid molecules encoding a chimeric antigen receptor (CAR) and a polypeptide comprising i) a ligand-binding extracellular domain (ECD); ii) an immune cell activation intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD. As provided herein, the particles may be viral particles. Therefore, this document also provides viral particles comprising a heterologous viral glycoprotein, a targeting portion, and one or more nucleic acid molecules encoding one or more heterologous molecules of interest, wherein the targeting portion comprises a polypeptide having the formula T-S1, where T is the target-binding domain and S1 is a stem portion, and wherein one or more nucleic acid molecules encode a chimeric antigen receptor (CAR) and a polypeptide comprising i) a ligand-binding extracellular domain (ECD); ii) an immune cell activation intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD. 。 In some embodiments, S1 comprises, for example, a mutant Fc polypeptide that can be linked to a transmembrane domain as provided herein. In some embodiments, S1 comprises a flexible polypeptide as provided herein. Mutant Fc polypeptides or flexible polypeptides can be incorporated into viral particles to help facilitate the targeting of viral particles to specific cell types.

[0181] Additionally, viral particles may contain VSV-G proteins that can be used, for example, to pseudotype viruses such as lentiviruses. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of vesicular stomatitis New Jersey virus strain, vesicular stomatitis Indiana virus strain, vesicular stomatitis Aragos virus strain, vesicular stomatitis Malaba virus strain, or vesicular stomatitis Carajás virus strain. Examples of such proteins are provided herein.

[0182] Pseudogenized viruses containing mutant VSV-G proteins (such as those provided herein) can be used in conjunction with targeting moieties to promote the fusion of the pseudotyped virus with specific cells or tissues based on the expression of the target on those cells or tissues. As provided herein, the targeting moieties can be linked to an Fc protein, which may refer to a stem protein containing a transmembrane domain to facilitate attachment of the targeting moieties to the viral surface. In some embodiments, the Fc protein contains Fc effector mutations, such as those provided herein.

[0183] Unless otherwise defined, all technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed embodiments pertain.

[0184] As used herein, unless the context clearly indicates otherwise, the term "a" means "at least one" or "one or more".

[0185] As used herein, the term “about” means an approximate index value, and small variations will not significantly affect the practice of the disclosed embodiments. When numerical limits are used, the “about” index value may vary by ±10% and remain within the range of the disclosed embodiments unless the context otherwise specifies. Furthermore, when the phrase “about x to y” is used, the term “about” modifies both x and y unless the context otherwise specifies, and can be used interchangeably with the phrase “about x to about y”.

[0186] As used herein, the interchangeable terms “individual” or “subject” or “patient” mean any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates such as humans.

[0187] As used herein, the terms “comprise” (and any form of inclusion, such as “comprise”, “comprises”, and “comprised”), “have” (and any form of having, such as “have” and “has”), “include” (and any form of inclusion, such as “includes” and “include”), or “contains” (and any form of containing, such as “contains” and “contains”) are inclusive or open-ended and do not exclude additional, unlisted elements or method steps. Any step or composition using the transitional phrase “comprise” or “comprising” may also be described as being identical to that described by the transitional phrase “consisting of” or “consists”.

[0188] As used herein, the term “contact” means bringing two elements together in an in vitro or in vivo system. For example, “contacting” a virus or vector described herein with an individual or patient or cells includes administering a virus to an individual or patient, such as a human, and, for example, introducing a compound into a sample containing cells or a purified preparation containing cells.

[0189] As used herein, the terms “fusion” or “linkage” when referring to proteins with different domains or heterologous sequences mean that the protein domains are part of the same peptide chain and are linked to each other by peptide bonds or other covalent bonds. Domains or segments may be directly linked or fused to each other, or another domain or peptide sequence may be between two domains or sequences, and such sequences will still be considered fused or linked to each other. In some embodiments, the various domains or proteins provided herein are directly linked or fused to each other, or adapter sequences such as the glycine / serine sequences described herein link two domains together.

[0190] "Disease" is a state of health in which the animal is unable to maintain homeostasis, and in which the animal's health continues to deteriorate if the disease is not treated. In contrast, an animal's "symptom" is a state of health in which the animal is able to maintain homeostasis, but in which the animal's health is worse than it would be without the symptom. A symptom, if left untreated, does not necessarily lead to a further decline in the animal's health.

[0191] The terms "effective amount" or "therapeutic effective amount" are used interchangeably herein and refer to the amount of a compound, formulation, material, or composition as described herein that effectively achieves a particular biological outcome or provides a therapeutic or preventative benefit. Such an outcome may include, but is not limited to, the amount of immune cell activation that, when administered to a mammal, results in a detectable level of immune cell activation compared to that detected in the absence of the composition. Immune responses can be readily assessed using a wide range of methods recognized in the art. Those skilled in the art will understand that the amount of the composition administered herein varies and can be readily determined based on many factors, such as the disease or symptom being treated, the age and health and physical condition of the mammal being treated, the severity of the disease, the specific compound administered, etc.

[0192] "Encoding" refers to the inherent characteristics of a specific nucleotide sequence in a polynucleotide (such as a gene, cDNA, or mRNA) for use as a template in biological processes to synthesize other polymers and macromolecules having defined nucleotide sequences (i.e., rRNA, tRNA, and mRNA) or defined amino acid sequences, and the resulting biological characteristics. Therefore, if the transcription and translation of the mRNA corresponding to a gene produces a protein in a cell or other biological system, then the gene encodes that protein. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is typically provided in the sequence listing, and the non-coding strand, which serves as a template for transcription of the gene or cDNA, can be referred to as the protein or other product encoding that gene or cDNA.

[0193] "Expression vector" refers to a vector containing a recombinant polynucleotide that includes an expression control sequence operatively linked to a nucleotide sequence to be expressed. The expression vector contains sufficient cis-acting elements for expression; other elements for expression may be provided by a host cell or in an in vitro expression system. Expression vectors include all expression vectors known in the art, such as clomids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., Sendai virus, lentivirus, retrovirus, adenovirus, and adeno-associated virus) incorporating recombinant polynucleotides.

[0194] As used in this article, the phrase “ex vivo” in relation to transducing, transfecting or transforming cells means transducing, transfecting or transforming cells outside of a subject, i.e., removing cells from a subject prior to transducing, transfecting or transforming such cells.

[0195] As used herein, “identity” refers to the subunit sequence identity between two polymer molecules, such as two nucleic acid or amino acid molecules, or such as two polynucleotide or polypeptide molecules. Two amino acid sequences are identical when they have the same residue at the same position; for example, if each position in two polypeptide molecules is occupied by arginine, then they are identical at that position. The degree of identity, or similarity, between two amino acid or two nucleic acid sequences in an alignment is usually expressed as a percentage. Identity between two amino acid or two nucleic acid sequences is a direct function of the number of matching or identical positions; for example, if half of the positions in two sequences are identical, then the two sequences are 50% identical; if 90% of the positions (e.g., 9 out of 10) are matching or identical, then the two amino acid sequences are 90% identical.

[0196] "Substantially identical" means that the polypeptide or nucleic acid molecule exhibits at least 50% identity with a reference amino acid sequence (e.g., any one of the amino acid sequences described herein) or nucleic acid sequence (e.g., any one of the nucleic acid sequences described herein). In some embodiments, such a sequence is at least 60%, 80%, or 85%, or 90%, 95%, or even 99% identical to the sequence used for comparison at the amino acid level or nucleic acid level. Other percentages of identity with respect to specific sequences are described herein.

[0197] Sequence identity can be measured / determined using sequence analysis software, such as the sequence analysis software package from Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, including the BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX programs. Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. Conserved substitutions typically include those within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. In exemplary methods for determining the degree of identity, the BLAST program can be used, where a probability score between e3 and e100 indicates closely related sequences. In some embodiments, sequence identity is determined using BLAST with default settings.

[0198] For the purposes of the embodiments provided herein, compositions comprising various proteins may, in some cases, contain amino acid sequences that have sequence identity with the amino acid sequences disclosed herein. Therefore, in some embodiments, depending on the specific sequence, the degree of sequence identity with the SEQ ID NO disclosed herein is preferably greater than 50% (e.g., 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater). Other identity percentages are also provided herein in addition to these percentages. Identity between peptides can be determined using an affine gap search with parameters gap opening penalty (12) and gap extension penalty = 1, as implemented in the MPSRCH program (Oxford Molecular).

[0199] These proteins, compared to publicly available proteins, may include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) conserved amino acid substitutions, where one amino acid is replaced by another amino acid with a related side chain. Genetically encoded amino acids are generally classified into four families: (1) acidic, i.e., aspartic acid and glutamic acid; (2) basic, i.e., lysine, arginine, and histidine; (3) nonpolar, i.e., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan; and (4) uncharged polar, i.e., glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes collectively classified as aromatic amino acids. Generally, substitution of a single amino acid within these families does not significantly affect biological activity. Proteins may have one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) single amino acid deletions relative to publicly available protein sequences. In addition to the publicly available protein sequence, the protein may also contain one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) insertions (e.g., each of 1, 2, 3, 4, or 5 amino acids).

[0200] As used in this article, the phrase “in vivo” in relation to transducing, transfecting, or transforming cells means transducing, transfecting, or transforming such cells in a subject without removing the cells from the subject prior to transducing, transfecting, or transforming them.

[0201] "Separated" means altered or removed from its natural state. For example, nucleic acids or peptides that are naturally present in living organisms are not "separated," but the same nucleic acids or peptides that are partially or completely separated from their natural counterparts are "separated." Separated nucleic acids or proteins can exist in a substantially purified form or in non-natural environments, such as, for example, host cells.

[0202] As used herein, “lentivirus” refers to a genus of the family Retroviridae capable of infecting non-dividing cells. Non-limiting examples of lentiviruses are HIV, SIV, and FIV. Vectors or virus-like particles derived from lentiviruses can be used to transduce cells and deliver genes or other molecules, enabling their expression in cells either in vitro (ex vivo) or in vivo.

[0203] As used herein, the term "modified" refers to an altered state or structure of a molecule or cell as presented herein. Molecules can be modified in many ways, including chemically, structurally, and functionally, such as through mutation, substitution, insertion, or deletion (e.g., internal deletion truncation). Cells can be modified by introducing nucleic acids or expressing heterologous proteins.

[0204] As used herein, the term "modulation" means mediating an increase or decrease in a subject's response level compared to the response level of a subject in the absence of treatment or the compound, and / or compared to the response level of other subjects who are otherwise identical but untreated. This term includes interfering with and / or influencing natural signals or responses, thereby mediating a beneficial therapeutic response in a subject (such as a human).

[0205] Unless otherwise specified, "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate to each other and encode the same amino acid sequence. Phrases encoding protein or RNA may also contain introns, to the extent that nucleotide sequences encoding proteins may contain introns in some forms.

[0206] The term "oligonucleotide" usually refers to short polynucleotides. It should be understood that when a nucleotide sequence is represented by a DNA sequence (i.e., A, T, C, G), this also provides the corresponding RNA sequence (i.e., A, U, C, G) in which "U" replaces "T".

[0207] "Parenteral" administration of the composition includes techniques such as subcutaneous (sc), intravenous (iv), intramuscular (im), or intrasternal injection or infusion.

[0208] As used herein, the term "polynucleotide" is defined as a nucleotide chain. Furthermore, nucleic acids are polymers of nucleotides. Therefore, as used herein, the terms "nucleic acid" and "polynucleotide" are interchangeable. As used herein, polynucleotides include, but are not limited to, all nucleic acid sequences obtained by any method available in the art, including, but not limited to, recombinant methods, i.e., cloning nucleic acid sequences from recombinant libraries or cell genomes using cloning techniques and PCR, as well as synthetic methods.

[0209] As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably and refer to compounds consisting of multiple amino acid residues covalently linked by peptide bonds. As used herein, the term refers to both short chains (which are also commonly referred to in the art as, for example, peptides, oligopeptides, and oligomers) and long chains (which are commonly referred to in the art as proteins, of which there are many types). “Polypeptide” includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, and so on. Polypeptides include natural peptides, recombinant peptides, synthetic peptides, or combinations thereof.

[0210] Unless otherwise explicitly stated, the terms “heteroviral structural protein” and “heteroviral glycoprotein” as used herein are synonymous and interchangeable. Therefore, unless otherwise explicitly stated, any implementation that refers to “heteroviral structural protein” should be understood to mean “heteroviral glycoprotein”, and vice versa.

[0211] As used herein, the term "pseudotyped" or "pseudotyped viral particle" refers to a viral particle carrying a glycoprotein derived from another enveloped virus or a viral vector encoding an envelope glycoprotein from a virus different from its parent virus. Therefore, the host range of a vector particle can be expanded or altered depending on the type of cell surface receptor used by the glycoprotein. For example, viruses can be pseudotyped using VSV-G mutant proteins as described herein.

[0212] As used herein with respect to antibodies, the term "specifically binding" refers to an antibody that recognizes a specific antigen but substantially does not recognize or bind to other molecules in the sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more species. However, this cross-species reactivity itself does not alter the antibody's specific classification. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen. However, this cross-reactivity itself does not alter the antibody's specific classification. In some cases, the term "specifically binding" or "specifically binding" may be used in relation to the interaction of an antibody, protein, or peptide with a second chemical substance to mean that the interaction depends on the presence of a specific structure on the chemical substance (e.g., an antigenic determinant or epitope); for example, the antibody recognizes and binds to a specific protein structure, rather than a protein in general. If an antibody is specific for epitope "A," then in a reaction containing labeled "A" and an antibody, the presence of a molecule containing epitope A (or free, unlabeled A) will reduce the amount of labeled A that binds to the antibody. In some implementations, the targeting portions of viral particles containing mutant VSV-G proteins or other viral structural proteins used for pseudotypening of viruses, as described herein, can bind specifically to their targets.

[0213] The term "subject" includes living organisms, including those that can elicit an immune response (e.g., mammals). As used herein, a "subject" or "patient" can be a human or a non-human mammal. Non-human mammals include, for example, livestock and pets such as sheep, cattle, pigs, dogs, non-human primates, cats, and rodents. In some embodiments, the subject is a human.

[0214] As used in this article, the term "therapeutic" refers to treatment and / or prevention. Therapeutic effects are achieved by suppressing, alleviating, or eradicating a disease state.

[0215] As used herein, the terms “transfected,” “transformed,” or “transduced” refer to the process of transferring or introducing exogenous nucleic acids into cells. “Transfected,” “transformed,” or “transduced” cells are cells that have been transfected, transformed, or transduced with exogenous nucleic acids. Cells include primary subject cells and their progeny. In some embodiments, transfection, transformation, or transduction occurs in vivo.

[0216] As used in this article, “treatment” means reducing the frequency or severity of at least one sign or symptom of a disease or condition experienced by the subject.

[0217] A "vector" is a composition of material containing isolated nucleic acids encoding proteins or peptides. Many vectors are known in the art, including but not limited to linear polynucleotides, plasmids, DNA, and RNA. Examples of viral vectors include, but are not limited to, Sendai virus vectors, adenovirus vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, etc.

[0218] "Carriers" or "delivery mediators" include viral particles, viruses, polylysine compounds, and liposomes that facilitate the transfer of nucleic acids into cells. Carriers or delivery mediators can also be used to deliver proteins or peptides into cells.

[0219] Scope: Throughout this disclosure, various aspects of the embodiments may be presented in a scope format. It should be understood that the scope format is for convenience and brevity only and should not be construed as an inflexible limitation. Therefore, the scope description should be considered as having specifically disclosed all possible sub-scopes and the individual values ​​within those scopes. For example, a scope such as 1 to 6 should be considered as having specifically disclosed sub-scopes such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and the individual numbers within those scopes, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the width of the scope. Unless otherwise expressly stated to the contrary, the disclosed scope also includes the endpoints of the scope.

[0220] Heterogeneous proteins

[0221] In some embodiments, peptides are provided as recombinant fusion proteins comprising: ii) a ligand-binding extracellular domain (ECD); iii) an immune cell-activating intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD.

[0222] As used herein, the term "immune cell activation intracellular domain (ICD)" refers to a domain that, when activated, can activate or increase the number of immune cells. In some embodiments, the immune cell activation ICD comprises the ICD of hGCSF-R or a mutant or fragment thereof. In some embodiments, the ICD of hGCSF-R comprises substitutions, insertions, or deletions. In some embodiments, the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111, or an amino acid sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it. In some embodiments, the hGCSF-R ICD does not comprise the sequence of SEQ ID NO: 110. In some embodiments, the hGCSF-R ICD contains the sequence of SEQ ID NO: 111 and does not contain amino acid residues 800-813, 775-813, 750-813, 725-813 or 716-813 of SEQ ID NO: 125 or fragments thereof located at the C-terminal residues of SEQ ID NO: 111.

[0223] In some embodiments, the hGCSF-R ICD contains the sequence of SEQ ID NO: 111 and amino acid residues 716-812 of SEQ ID NO: 125 or a fragment of the residues at the C-terminus of SEQ ID NO: 111, provided that the ICD does not contain a continuous fragment of amino acid residues 716-813 of SEQ ID NO: 125.

[0224] As used herein, the term "ligand-binding extracellular domain (ECD)" refers to a protein domain that can bind to a ligand and can be used to transmit signals via the ICD. This signal can be a signal that enhances cell activation or increases the rate of cell division to increase cell number. The ligand can be naturally occurring, synthetic, or prepared recombinantly. For example, G-CSF (granulocyte colony-stimulating factor) or human growth hormone (HGH) can be used as ligands that can bind to the ECD to transmit signals. For example, growth hormone can be used as a ligand that can bind to the ECD, which in turn can bind to HGH. Additionally, in some implementations, lenograstim (Granocyte), filgrastim (Neupogen, Zarcio, Nivestim, Accofil), long-acting (pegylated) filgrastim (pegfilgrastim, Neulasta, Pelmeg, Ziextenco), or lipegfilgrastim (Lonquex) can be used as ligands that can bind to G-CSF, which in turn binds to HGH. Examples of ECDs provided in this article include, but are not limited to, G-CSFR or hGHR. Without being bound by any particular theory, when these drugs are administered to patients expressing these peptides, T cells will proliferate (increase in number) and lead to a greater response against the tumor.

[0225] In some embodiments, the ECD is a GCSF-R ECD. In some embodiments, the GCSF-R ECD is a human GCSF-R ECD. In some embodiments, the human GCSF-R ECD contains the amino acid sequence of SEQ ID NO: 107 or SEQ ID NO: 108.

[0226] In some embodiments, the ECD comprises an ECD of the human growth hormone receptor (hGH-R) or a mutant or fragment thereof. In some embodiments, the ECD of the human growth hormone receptor (hGH-R) comprises an amino acid sequence encoded by exons 1-7 of hGH-R or a mutant or fragment thereof. In some embodiments, the ECD of hGH-R comprises a mutation of C259R. In some embodiments, the ECD of hGH-R comprises the amino acid sequence of SEQ ID NO: 114 or SEQ ID NO: 115, or an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it. In some embodiments, the ECD of hGH-R comprises the amino acid sequence of SEQ ID NO: 114 or SEQ ID NO: 115 excluding the leader sequence, or an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

[0227] In some embodiments, the ICD comprises an ICD of glycoprotein 130 (“gp130 ICD”) or a variant or fragment thereof. In some embodiments, the gp130 ICD comprises a fragment of gp130 ICD. In some embodiments, the gp130 ICD fragment comprises a Jak1 binding site and / or an SHP-2 / STAT3 binding site. A non-limiting example is the amino acid sequence of SEQ ID NO: 122. Another non-limiting example of gp130 ICD or a fragment thereof includes the amino acid sequence of the gp130 ICD of construct G, which is described in further detail herein.

[0228] In some embodiments, the ICD contains a fragment of the GCSF-R ICD. In some embodiments, the GCSF-R ICD fragment does not contain STAT3, SH3, and / or SHC domains. In some embodiments, the GCSF-R ICD fragment contains Jak1 and / or STAT3 binding sites. In some embodiments, the ICD contains IL-2RβICD or a fragment or variant thereof. In some embodiments, IL-2RβICD is a fragment of IL-2RβICD. In some embodiments, IL-2RβICD or a fragment thereof contains an SHC or STAT5 binding domain. In some embodiments, the ICD contains IL-7RαICD or a fragment or variant thereof. In some embodiments, IL-7RαICD is a fragment of IL-7RαICD. In some embodiments, the ICD contains IL-7RαICD or a fragment thereof, containing STAT5 and / or PI3K binding domains. In some embodiments, the ICD contains a gp130 ICD or a variant or fragment thereof linked to IL-2RβICD or a fragment thereof. A non-limiting example of IL-2RβICD or a fragment thereof is the amino acid sequence of SEQ ID NO: 123. In some embodiments, the ICD comprises an hCCSF-R ICD or a variant or fragment thereof linked to IL-7RαICD or a fragment thereof. A non-limiting example of IL-7RαICD or a fragment thereof is the amino acid sequence of SEQ ID NO: 118. Another non-limiting example of IL-7RαICD or a fragment thereof includes the amino acid sequence of IL-7RαICD of construct E, which is described in further detail herein.

[0229] In some embodiments, the transmembrane domain comprises a GCSF-R, gp130, or hGH-R transmembrane domain. In some embodiments, the transmembrane domain comprises an hGCSF-R transmembrane domain or a variant thereof. In some embodiments, the transmembrane domain comprises a gp130 transmembrane domain or a variant thereof. Examples of gp130 transmembrane domains include, but are not limited to, the amino acid sequence of SEQ ID NO: 121. In some embodiments, the transmembrane domain comprises an hGH-R transmembrane domain or a variant thereof.

[0230] In some embodiments, the peptide comprises an ECD as a GCSFR-ECD or a variant thereof (e.g., as shown herein and above); a transmembrane domain as a GCSF-R transmembrane domain or a variant thereof (e.g., as shown herein and above); and an immune-activated ICD as a GCSF-R ICD or a variant thereof (e.g., as shown herein and above).

[0231] In some embodiments, the peptide comprises an ECD as hGH-R-ECD or a variant thereof (e.g., as shown herein and above); a transmembrane domain as a GCSF-R transmembrane domain or a variant thereof (e.g., as shown herein and above); and an immune-activating ICD as a GCSF-R ICD or a variant thereof (e.g., as shown herein and above).

[0232] In some embodiments, the peptide comprises an ECD as hGH-R-ECD or a variant thereof (e.g., as shown herein and above); a transmembrane domain as gp130 transmembrane domain or a variant thereof (e.g., as shown herein and above); and an immune-activating ICD comprising gp130 ICD or a variant thereof (e.g., as shown herein and above) and IL-2Rβ ICD or a variant thereof (e.g., as shown herein and above).

[0233] In some embodiments, the peptide comprises an ECD as hGH-R-ECD or a variant thereof (e.g., as shown herein and above); a transmembrane domain as a GCSF-R transmembrane domain or a variant thereof (e.g., as shown herein and above); and an immune-activating ICD comprising a GCSF-R ICD or a variant thereof (e.g., as shown herein and above) and an IL-7Rα ICD or a variant thereof (e.g., as shown herein and above).

[0234] In some embodiments, the polypeptide comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124, or a variant thereof. In some embodiments, the variant comprises a polypeptide having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124.

[0235] Examples of these domains or sequences are provided in this specification.

[0236] The peptides with different domains described in this section are different from so-called chimeric antigen receptors. Chimeric antigen receptors can coexist with these peptides in the same cell, but those skilled in the art would not consider the peptide to be a chimeric antigen receptor. Therefore, in some embodiments, the peptide does not contain an antigen-binding domain. For example, in some embodiments, the peptide does not contain an antigen-binding domain in the form of scFv, Fab, or a single-domain antibody. Other antigen-binding domains are provided herein, and in some embodiments, the peptide does not contain any form of antigen-binding domain. Those skilled in the art would not consider the ligand ECD domain to be the same as the antigen-binding domain. Examples of domains that a peptide may not contain are also described in Guedan et al., Engineering and Design of Chimeric Antigen Receptors, Molecular Therapy: Methods & Clinical Development, Vol. 12, March 2019, pp. 145-156, which is incorporated herein by reference in its entirety.

[0237] In some embodiments, these peptides do not contain an ECD containing a binding domain for binding tumor antigens. In some embodiments, the peptides do not contain a co-stimulatory domain. In some embodiments, the peptides do not contain a CD3ζ signaling domain. In some embodiments, the peptides do not contain a CD28ζ signaling domain. In some embodiments, the peptides do not contain an immune receptor tyrosine-based activation motif (ITAM). In some embodiments, the peptides do not contain a co-stimulatory intracellular domain. In some embodiments, the peptides do not contain a 4-1BB signaling or co-stimulatory intracellular domain. In some embodiments, the peptides do not contain a CD28 signaling or co-stimulatory intracellular domain. In some embodiments, the peptides do not contain an inducible T cell co-stimulatory factor (ICOS) intracellular domain.

[0238] In some implementations, the polypeptide does not contain the CH2, CH3, CD28, or CD8α hinge regions.

[0239] In some implementations, peptides containing G-CSFR-containing ECDs or variants thereof (such as those provided herein) do not contain an antigen-binding domain.

[0240] These polypeptides can be incorporated into any of the particles provided herein. Particles containing polypeptides are provided according to some embodiments. In some embodiments, the particles are vectors (e.g., viruses, liposomes, plasmids, etc.) or cells. In some embodiments, the particles are viruses or viral vectors. In some embodiments, the viral vectors are lentiviral vectors, AV vectors, AAV vectors, etc. In some embodiments, the viral vector is a pseudotyped viral vector. Non-limiting examples of pseudotyped viral vectors are provided herein, but they can be pseudotyped using any viral structural protein. In some embodiments, the particles also contain chimeric antigen receptors, such as chimeric antigen receptors containing antigen-binding domains that bind to tumor antigens.

[0241] This document also provides nucleic acid molecules encoding chimeric antigen receptors (CARs), peptides, or combinations thereof. In some embodiments, the CAR is as provided herein. In some embodiments, the CAR and peptide are encoded by distinct nucleic acid molecules. In some embodiments, the CAR and peptide are encoded by the same nucleic acid molecule. Methods for encoding multiple proteins of interest by a single nucleic acid molecule are known in the art, and any such methods are within the scope of this disclosure. In some embodiments, a single nucleic acid molecule encodes the CAR and peptide as a bicistronic molecule, wherein the CAR and peptide are separated by a cleavable peptide linker. Cleavable peptide linkers are known in the art, and any such cleavable linker is within the scope of this disclosure. In some embodiments, the cleavable peptide linker is a 2A peptide linker. In some embodiments, the nucleic acid molecule encodes a bicistronic construct, wherein from 5' to 3', the nucleic acid molecule encodes the CAR, a cleavable linker (e.g., a 2A linker), and the peptide. In some embodiments, the nucleic acid molecule encodes a bicistronic construct, wherein from 5' to 3', the nucleic acid molecule encodes the peptide, a cleavable linker (e.g., a 2A linker), and the CAR. In some embodiments, particles comprising the nucleic acid molecule are provided. In some embodiments, the particle is a vector (e.g., a virus, liposome, plasmid, etc.) or a cell. In some embodiments, the particle is a virus or a viral vector. In some embodiments, the viral vector is a lentiviral vector, an AV vector, an AAV vector, etc. In some embodiments, the viral vector is a pseudotyped viral vector. This document provides non-limiting examples of pseudotyped viral vectors, but pseudotyped viral vectors can be used with any viral structural protein.

[0242] Viral particles containing heterologous viral glycoproteins and targeting components

[0243] In some embodiments, a viral particle comprising a heterologous viral glycoprotein and a targeting portion is provided. In some embodiments, the targeting portion comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stem portion. In some embodiments, S1 comprises a variant Fc protein, wherein the variant Fc protein comprises a transmembrane domain, such as, but not limited to, a CD8 or CD28 transmembrane domain. Thus, in some embodiments, the stem portion S1 comprises an N-terminal to C-terminal orientation (transmembrane domain) of the variant Fc. In some embodiments, the variant Fc protein comprises an effector mutation, wherein the effector mutation inhibits the interaction between the Fc protein and Fc-interacting proteins, such as FcγR, C1q, FcRβ, or FcRn.

[0244] In some embodiments, the S1 stem portion is attached to the surface of the viral particle via a transmembrane domain. In some embodiments, the variant Fc protein is a variant of the IgG1 Fc, IgG2 Fc, or IgG4 Fc protein. In some embodiments, the mutant Fc protein comprises a variant of the sequence SEQ ID NO: 26 (IgG1 Fc), SEQ ID NO: 27 (IgG2 Fc), or SEQ ID NO: 28 (IgG4 Fc).

[0245] In some embodiments, the mutant Fc protein is the variant IgG1 Fc protein (SEQ ID NO: 26). In some embodiments, the variant IgG1 Fc protein comprises one or more mutations corresponding to those mutations selected from the group consisting of: L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26 according to the Kabat EU number index, as described in Edelman, GM et al., “The covalent structure of anentire gammaG immunoglobulin molecule.” Proceedings of the National Academy of Sciences of the United States of America, Vol. 63, 1 (1969): 78-85. doi:10.1073 / pnas.63.1.78, which is incorporated herein by reference in its entirety. The mutations L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26 may or may not be present, and the mutations may be combined in any combination. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A and L235A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to N297A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to P329G of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, and P329G of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to I253A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H310A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H435A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to I253A, H310A, and H435A of SEQ ID NO: 26. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26.

[0246] In some embodiments, the variant Fc protein comprising the variant IgG1 Fc protein comprises a truncated IgG1 Fc sequence. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the IgG1 Fc sequence. In some embodiments, the variant Fc protein comprising the variant IgG1 Fc protein comprises a truncated SEQ ID NO: 26. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of SEQ ID NO: 26. In some embodiments, truncating includes the deletion of amino acids from the N-terminus of SEQ ID NO: 26. In some embodiments, truncating includes the deletion of amino acids from the C-terminus of SEQ ID NO: 26. In some embodiments, truncating includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 26. In some embodiments, the truncated SEQ ID NO: 26 comprises the amino acid sequence of SEQ ID NO: 103:

[0247] EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 103)

[0248] In some embodiments, the variant IgG1 Fc protein contains one or more mutations relative to SEQ ID NO: 103, selected from the group consisting of L19A, L20A, N82A, P114G, I38A, H95A, and H220A. It should be understood that positions L19, L20, N82, P114, I38, H95, and H220 refer only to SEQ ID NO: 103. Those skilled in the art will readily recognize that positions L234, L235, N297, P329, I253, H310, and H435, numbered according to Kabat's EU numbering system, correspond to positions L19, L20, N82, P114, I38, H95, and H220 of SEQ ID NO: 103, respectively. The mutations L19A, L20A, N82A, P114G, I38A, H95A, and H220A in SEQ ID NO: 103 may or may not be present, and the mutations may be combined in any combination. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L19A and L20A of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to N82A of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to P114G of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L19A, L20A, N82A, and P114G of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to I38A of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H95A of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to H220A of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to I38A, H95A, and H220A of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains mutations corresponding to L19A, L20A, N82A, P114G, I38A, H95A, and H220A of SEQ ID NO: 103. In some embodiments, the variant IgG1 Fc protein contains a mutation corresponding to each of L18A, L19A, N82A, P114G, I38A, H95A, and H220A of SEQ ID NO: 103.Exemplary variants of IgG1 protein comprising each of L18A, L19AN82A, P114G, I38A, H95A, and H220A are shown in the amino acid sequence of SEQ ID NO: 104 below.

[0249] EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMASRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLAQDWLNGKEYKCKVSNKALGAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNAYTQKSLSLSPGK (SEQ ID NO: 104)

[0250] In some embodiments, the mutant Fc protein is the variant IgG2 Fc protein (SEQ ID NO: 27). In some embodiments, the variant IgG2 Fc protein comprises one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A according to the Kabat EU number index, because those positions correspond to SEQ ID NO: 27. Any of the mutations N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 27 may or may not be present, and the mutations may be combined in any combination. In some embodiments, the variant IgG2 Fc protein comprises a mutation corresponding to N297A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises a mutation corresponding to P329G of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein comprises mutations corresponding to both N297A and P329G of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to I253A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to H310A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains the mutation corresponding to H435A of SEQ ID NO: 27. In some embodiments, the variant IgG2 Fc protein contains mutations corresponding to I253A, H310A, and H435A of SEQ ID NO: 27.

[0251] In some embodiments, the variant Fc protein comprising the variant IgG2 Fc protein comprises a truncated IgG2 Fc sequence. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the IgG2 Fc sequence. In some embodiments, the variant Fc protein comprising the variant IgG2 Fc protein comprises a truncated SEQ ID NO: 27. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of SEQ ID NO: 27. In some embodiments, truncating includes the deletion of amino acids from the N-terminus of SEQ ID NO: 27. In some embodiments, truncating includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 27.

[0252] In some embodiments, the mutant Fc protein is the variant IgG4 Fc protein (SEQ ID NO: 28). In some embodiments, the variant IgG4 Fc protein comprises one or more mutations selected from the group consisting of: S228P, L235E, N297A, P329G, I253A, H310A, and H435A according to the Kabat EU number index, because those positions correspond to SEQ ID NO: 28. Any of the mutations S228P, L235E, N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 28 may or may not be present, and the mutations may be combined in any combination. In some embodiments, the variant IgG4 Fc protein comprises the mutation corresponding to S228P of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein comprises the mutation corresponding to L235E of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to N297A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to P329G of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains mutations corresponding to S228P, L235E, N297A, and P329G of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to I253A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to H310A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains a mutation corresponding to H435A of SEQ ID NO: 28. In some embodiments, the variant IgG4 Fc protein contains mutations corresponding to I253A, H310A, and H435A of SEQ ID NO: 28.

[0253] In some embodiments, the variant Fc protein comprising the variant IgG4 Fc protein comprises a truncated IgG4 Fc sequence. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the IgG4 Fc sequence. In some embodiments, the variant Fc protein comprising the variant IgG4 Fc protein comprises a truncated SEQ ID NO: 28. Truncating may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of SEQ ID NO: 28. In some embodiments, truncating includes the deletion of amino acids from the N-terminus of SEQ ID NO: 28. In some embodiments, truncating includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 28.

[0254] In some embodiments, the stem portion S1 comprising the variant Fc protein is given by the formula L1-Fc-L2-X1, where L1 is a linker or is absent; Fc is the variant Fc protein; L2 is a linker or is absent; and X1 is a polypeptide comprising a transmembrane domain. Therefore, the targeting portion comprising the formula T-S1 can also be given by the formula T-L1-Fc-L2-X1, where T is a target-binding domain; L1 is a linker or is absent; Fc is the variant Fc protein; L2 is a linker or is absent; and X1 is a polypeptide comprising a transmembrane domain. Therefore, it should be understood that in some embodiments, the stem portion S1 can be given by the formula L1-Fc-L2-X1. In some embodiments, the target-binding domain T is as provided herein. In some embodiments, the variant Fc protein is as provided herein.

[0255] In some embodiments, L1 and L2 are each independently a peptide linker. In some embodiments, the peptide linker includes (GGGGA). n (SEQ ID NO: 54), (GGGGS) n (SEQ ID NO: 55), (EAAAK) n (SEQ ID NO: 73), A(EAAAK) n A (SEQ ID NO: 74), (XP) n(SEQ ID NO: 75) (where X is Ala, Lys, or Glu), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESKST (SEQ ID NO: 58), AEAAAKEAAAKA (SEQ ID NO: 76) or combinations thereof, wherein each n is independently 1-5. In some embodiments, each n is independently 1. In some embodiments, each n is independently 2. In some embodiments, each n is independently 3. In some embodiments, each n is independently 4. In some embodiments, each n is independently 5. In some embodiments, each n is independently greater than 5. In some embodiments, L1 is not present. In some embodiments, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1-5. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 2. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 3. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 4. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 5. In some implementations, L1 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently greater than 5. In some embodiments, L2 is absent. In some embodiments, L2 is (GGGGA).n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1-5. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 1. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 2. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 3. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently 4. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO:55), where each n is independently 5. In some implementations, L2 is (GGGGA). n (SEQ ID NO: 54) or (GGGGS) n (SEQ ID NO: 55), where each n is independently greater than 5.

[0256] In some implementations, X1 includes a formula ECD-T M -ICD polypeptides, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M It is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the targeting portion into the viral particle envelope, or it may not exist. Therefore, in some embodiments, the targeting portion comprising formula T-S1, which may also be given by formula T-L1-Fc-L2-X1, may also be given by formula T-L1-Fc-L2-ECD-T. M -ICD is given, where T is the target-binding domain; L1 is the linker or absent; Fc is the variant Fc protein; L2 is the linker or absent; ECD is the extracellular domain or fragment of a cell surface protein, or absent; T MIt is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the target portion into the viral particle envelope, or it may not exist. Therefore, it should be understood that in some embodiments, the stem portion S1 may be composed of the formula L1-Fc-L2-ECD-T M -ICD provides this information.

[0257] In some embodiments, the stem portion S1 does not contain a variant Fc region. In some embodiments, the stem portion S1 is given by formula L3-X1, where L3 is a flexible peptide linker and X1 is a polypeptide containing a transmembrane domain as provided herein. Therefore, in some embodiments, the targeting portion containing formula T-S1 may also be given by formula T-L3-X1, where T is a target-binding domain, L3 is a flexible peptide linker, and X1 is a polypeptide containing a transmembrane domain as provided herein. Therefore, it should be understood that in some embodiments, the stem portion S1 may be given by formula L3-X1. In some embodiments, the S1 stem portion is attached to the surface of the viral particle via a transmembrane domain.

[0258] In some embodiments, the flexible peptide linker L3 can be any flexible peptide linker. In some embodiments, L3 is selected from the group consisting of flexible linkers, including but not limited to (GGGGA). n (SEQ ID NO: 54), (GGGGS) n (SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESSKST (SEQ ID NO: 58), or any combination thereof, wherein each n is independently an integer selected from 1 to 4. In some embodiments, each n is independently an integer selected from 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, or 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, each n is independently greater than 10. In some embodiments, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 1. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 2. In some implementations, L3 is (GGGGA). n(SEQ ID NO:54) and n is 3. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 4. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 5. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 6. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 7. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 8. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 9. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 10. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is greater than 10. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 1. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 2. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 3. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 4. In some implementations, L3 is (GGGGS). n (SEQ ID NO:55) and n is 5. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 6. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 7. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 8. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 9. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 10. In some implementations, L3 is (GGGGS).n (SEQ ID NO: 55) and n is greater than 10. In some embodiments, L3 is GSAGSAAGSGEF (SEQ ID NO: 56). In some embodiments, L3 is KESGSVSSEQLAQFRSLD (SEQ ID NO: 57). In some embodiments, L3 is EGKSSGSGSESKST (SEQ ID NO: 58).

[0259] In some embodiments, the flexible peptide linker L3 can be any flexible peptide linker. In some embodiments, L3 is selected from the group consisting of flexible linkers, including but not limited to (GGGGA). n (SEQ ID NO: 54), (GGGGS) n (SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESSKST (SEQ ID NO: 58), or any combination thereof, wherein each n is independently an integer selected from 1 to 4. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 1. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 2. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 3. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 4. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 1. In some implementations, L3 is (GGGGS). n (SEQ ID NO:55) and n is 2. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 3. In some implementations, L3 is (GGGGS). n(SEQ ID NO: 55) and n is 4. In some embodiments, L3 is GSAGSAAGSGEF (SEQ ID NO: 56). In some embodiments, L3 is KESGSVSSEQLAQFRSLD (SEQ ID NO: 57). In some embodiments, L3 is EGKSSGSGSESKST (SEQ ID NO: 58).

[0260] In some implementations, L3 is selected from the group consisting of flexible joints, including but not limited to (GGGGA). n (SEQ ID NO:54), (GGGGS) n (SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESSKST (SEQ ID NO: 58), or any combination thereof, wherein each n is independently an integer selected from 1, 2, or 4. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 4. In some embodiments, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 1. In some implementations, L3 is (GGGGA). n (SEQ ID NO: 54) and n is 2. In some implementations, L3 is (GGGGA). n (SEQ ID NO:54) and n is 4. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 1. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 2. In some implementations, L3 is (GGGGS). n (SEQ ID NO: 55) and n is 4. In some embodiments, L3 is GSAGSAAGSGEF (SEQ ID NO: 56). In some embodiments, L3 is KESGSVSSEQLAQFRSLD (SEQ ID NO: 57). In some embodiments, L3 is EGKSSGSGSESKST (SEQ ID NO: 58).

[0261] In some implementations, X1 includes a formula ECD-T M -ICD polypeptides, wherein ECD is an extracellular domain of a cell surface protein or a fragment thereof, or is absent; T MIt is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the targeting portion into the viral particle envelope, or it is absent. Therefore, in some embodiments, the targeting portion comprising formula T-S1, which may also be given by formula T-L3-X1, may also be given by formula T-L3-ECD-T. M -ICD is given, where T is the target-binding domain, L3 is the flexible peptide linker, and ECD is the extracellular domain of a cell surface protein or a fragment thereof, or it may not exist; T M It is a transmembrane domain of a transmembrane protein; and the ICD is an intracellular domain of a protein or a protein that promotes the incorporation of the target portion into the viral particle envelope, or it may not exist. Therefore, it should be understood that in some embodiments, the stem portion S1 may be derived from formula L3-ECD-T. M -ICD provides this information.

[0262] In some embodiments, the ECD is absent. In some embodiments, the ECD can be any suitable extracellular domain or a fragment thereof. In some embodiments, the ECD originates from a different protein than the transmembrane domain. The ECD domain can be the entire ECD domain or a fragment thereof. In some embodiments, the ECD domain is a CD8 or CD28 ECD domain or a fragment thereof. In some embodiments, the ECD domain is a CD8 ECD domain or a fragment thereof. In some embodiments, the CD8 ECD domain comprises a polypeptide of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the CD8 ECD domain consists of or is substantially composed of a polypeptide of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain comprises a polypeptide of 25-45 amino acids in length. In some embodiments, the ECD comprises at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the peptide of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain is a CD28 ECD domain or a fragment thereof. In some embodiments, the CD28 ECD domain comprises a polypeptide of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the CD28 ECD domain is composed of or substantially composed of the polypeptide of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the ECD domain comprises a polypeptide of 25-45 amino acids in length. In some embodiments, the ECD contains at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the peptide of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO:60).

[0263] In some implementations, T M It can be any suitable transmembrane domain or fragment thereof. In some embodiments, T MThe structural domain is CD8 or CD28 T M A domain or a fragment thereof. In some implementations, T M The structural domain is CD8 T M A domain or a fragment thereof. In some implementations, CD8 T M The domain contains a polypeptide of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, CD8 T M The domain is composed of or substantially composed of a polypeptide of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M The polypeptide contains at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the peptide identical to that of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M The structural domain is CD28T M A domain or a fragment thereof. In some implementations, CD28 T M The domain comprises a polypeptide of the form FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, CD28 T M The domain is composed of or substantially composed of a polypeptide of the form FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, T M The polypeptide contains at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the same peptide as FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62).

[0264] In some implementations, T M The domain originates from the same protein as ECD. In some implementations, T M The domain originates from a protein different from ECD. In some implementations, T M The structural domain is CD8 or CD28 T M A structural domain or a fragment thereof, and the ECD structural domain is a CD8 or CD28 ECD structural domain or a fragment thereof. In some embodiments, T MThe peptide with the structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The peptide with the structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, T M The peptide with the structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62), and the ECD structural domain is at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The peptides of FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62) have at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in terms of their structural domains, and the peptides of KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60) have at least or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in terms of their ECD domains.

[0265] In some embodiments, the transmembrane domain is linked to the intracellular domain (ICD) of a cellular transmembrane protein or a fragment thereof. In some embodiments, the ICD is absent. In some embodiments, the ICD originates from a protein that is the same as or different from the TM domain. In some embodiments, the ICD contains an Env incorporation motif. An Env incorporation motif is a molecule, such as a polypeptide, that can facilitate protein incorporation into the viral envelope. A non-limiting example of an Env incorporation motif is a polypeptide containing the amino acid sequence NRVRQGYS (SEQ ID NO: 63). This is a non-limiting example, and other peptide sequences, such as, but not limited to, GGTETSQVAPA (SEQ ID NO: 64), can be used. In some embodiments, the Env incorporation motif contains the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 64, or a combination thereof. In some embodiments, the Env incorporation motif contains the amino acid sequence of SEQ ID NO: 63. In some embodiments, the Env incorporation motif contains the amino acid sequence of SEQ ID NO: 64.

[0266] In some implementations, the target-binding domain "T" can be any polypeptide or polynucleotide that can be used to bind to the desired target. In some implementations, T is any polypeptide, polynucleotide, or fragment thereof that binds to: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR α, TCR β, TCR γ, TCR δ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3; glycosylated CD43 epitopes expressed on acute leukemia or lymphoma but not on hematopoietic progenitor cells; glycosylated CD43 epitopes expressed on non-hematopoietic cancers; A kinase anchoring protein 4 (AKAP-4); adrenaline receptor β3 (ADRB3); AFP; anaplastic lymphoma kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie 2) Anti-desmosome core protein 1 (Dsg1) autoantibody; anti-desmosome core protein 3 (Dsg3) autoantibody; B7H3 (CD276); biotin; bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; cancer / testis antigen 1 (NY-ESO-1); cancer / testis antigen 2 (LAGE-la); carbonic anhydrase IX (CA1X); carcinoembryonic antigen (CEA); CCCTC binding factor (zinc finger protein)-like (BORIS or imprinted site regulator sibling factor); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32 (FCG) R2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGLll; CD200R; CD276 / B7H3; CD300 molecular-like family member f (CD300LF); CDH1-CD324; CDH6; CDH17; CDH19; Chromosome X open reading frame 61 (CXORF61); Sealing protein 6 (CLDN6); Sealing protein 18.2 (CLD18A2 or CLDN18A.2); CMV pp65; C-MYC epitope tag; Cripto; CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24); CSF2RA (GM-CSFR-α); C-type lectin domain family 12 member A (CLEC12A); C-type lectin-like molecule-1 (CLL-1 or CLECL1); Cyclin B1; Cytochrome P450 IB 1 (CYP1B 1); DLL3; EBV-EBNA3c;Contains EGF-bke module 2 (EMR2) containing mucin-like hormone receptor-like receptors; Elongation factor 2 mutant (ELF2M); liver glycoside B2; liver glycoside type A receptor 2 (EphA2); epidermal growth factor receptor (EGFR); epidermal growth factor receptor variant III (EGFRviii); epithelial cell adhesion molecule (EPCAM); ERG; ETS translocation variant gene 6 located on chromosome 12p (ETV6-AML); Fc fragment of IgA receptor (FCAR or CD89); Fc receptor-like 5 (FCRL5); fibroblast activating protein α (FAP); FITC; Fms FLT3; folate receptor α (FRa or FR1); folate receptor β (FRb); follicle-stimulating hormone receptor (FSHR); Fos-associated antigen 1; fucosylated GM1; G protein-coupled receptor class C5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; ganglioside G2 (GD2); ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(l-4)bDGlcp(ll)Cer); ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l- 4) bDGlcp(ll)Cer); GD3; GFRα4; Glycoprotein 100 (gplOO); Phosphatidylinositol proteoglycan-3 (GPC3); Gonadotropin receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylate cyclase C (GCC); Mutant heat shock protein 70-2 (mut hsp70-2); Hepatitis A virus cell receptor 1 (HAVCR1); GloboH glycosylceramide hexasaccharide moiety (GloboH); High molecular weight melanoma-associated antigen (HMWMAA); HIV1 envelope glycoprotein; HLA; HLA-DOA; HLA-A; HLA-A2; HLA-B; HLA-C; HLA-DM; HLA-DOB; HLA-DP; HLA-DQ; HLA-DR; HLA-G; HTLV1-Tax; Human papillomavirus E6 (HPV) E6); Human papillomavirus E7 (HPV E7); Human telomerase reverse transcriptase (hTERT); IgE; IL13Ra2; IL11Ra; Immunoglobulin λ-like polypeptide 1 (IGLL1); Influenza A hemagglutinin (HA); Insulin-like growth factor 1 receptor (IGF-I receptor); Interleukin 11 receptor α (IL-11Ra); Interleukin 13 receptor subunit α-2 (IL-13Ra2 or CD213A2); Intestinal carboxylesterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; podin; Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2).Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); Luteinizing hormone receptor (LHR); Lewis (Y) antigen; Lews Ag; Livl; Locus K9 (LY6K); Low-conductivity chloride channel; Lymphocyte antigen 6 complex; Lymphocyte antigen 75 (LY75); Lymphocyte-specific protein tyrosine kinase (LCK); Breast differentiation antigen (NY-BR-1); T-cell recognized melanoma antigen 1 (MelanA or MARTI); Melanoma-associated antigen 1 (MAGE-A1); Melanoma cancer testis antigen-1 (MAD-CT-1); Melanoma cancer testis antigen-2 (MAD-CT-2); Melanoma cell apoptosis inhibitor (ML-IAP); Mesothelin; MPL; Cell surface-associated mucin 1 (MUC1); N-acetylglucosamine transferase V (NA17); Ligin-4; Neural cell adhesion molecule (NCAM); NKG 2D; NYBR1; O-acetyl-GD2 ganglioside (OAcGD2); olfactory receptor 51E2 (OR51E2); oncogene fusion protein composed of breakpoint cluster region (BCR) and Abelson murine leukemia virus oncogene homolog 1 (Abl) (bcr-abl); P53 mutant; pairing box protein Pax-3 (PAX3); pairing box protein Pax-5 (PAX5); pan-connecting protein 3 (PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor β (PDGFR-β); polysialic acid; proapocrine-binding protein sp32 (OY-TES1); prostate enzyme; prostate cancer tumor antigen-1 (PCT) A-1 or galactohemagglutinin 8); prostate stem cell antigen (PSCA); prostate-specific membrane antigen (PSMA); prostatic acid phosphatase (PAP); Prostein; serine protease 21 (Testisin or PRSS21); proteasome macropain subunit β Type 9 (LMP2); PTK7; RasG12V; Ras homolog family member C (RhoC); Rat sarcoma (Ras) mutant; Receptor to advanced glycation end products (RAGE-1); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Receptor tyrosine protein kinase ERBB2 or Her-22 / neu; Renal universal protein 1 (RU1); Renal universal protein 2 (RU2); Sarcoma translocation breakpoint; Serine 2 (TMPRSS2) ETS fusion gene; Sialoyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialic acid Lewis antigen); Sperm protein 17 (SPA17); T cell recognized squamous cell carcinoma antigen 3 (SART3); Stage-specific embryonic antigen-4 (SSEA-4); STEAP1; Survival protein; Synovial sarcoma X breakpoint 2 (SSX2).TCR γ-substituted reading frame protein (TARP); TCR-β1 chain; TCR-β2 chain; TCR-δ chain; TCR-γ chain; TCRγ-δ; telomerase; TGFβR2; antigen recognized by TNT antibody; thyroid-stimulating hormone receptor (TSHR); Timol / HVCR1; tissue factor 1 (TF1); Tn ag; Tn antigen ((Tn) (Ag) or (GalNAca-Ser / Thr)); TNF receptor family member B cell maturation (BCMA); transglutaminase 5 (TGS5); transmembrane protease; TROP2; tumor endothelial marker 1 (TEM1 / CD248); tumor endothelial marker 7 associated (TEM7R); tumor protein p53 (p53); tumor-associated glycoprotein 72 (TAG72); tyrosinase; tyrosinase-associated protein 2 (TRP-2); urolytic protein 2 (UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein (WT1); or X antigen family member 1A (XAGE1). In some embodiments, the target domain "T" binds to CD7. In some embodiments, the target-binding domain "T" binds to CD8. In some embodiments, the target-binding domain "T" is an antibody. It should be understood that, in the context of this disclosure, "antibody" refers not only to a "complete" antibody comprising two identical heavy chains, two identical light chains, and two antigen-binding fragments, but also to any isotype of antibody, antibody fragment (including but not limited to Fab, Fv, scFv, and Fd fragments), chimeric antibody, humanized antibody, single-chain antibody (scAb), single-domain antibody (dAb), single-domain heavy chain antibody, single-domain light chain antibody, bispecific antibody, multispecific antibody, and fusion protein comprising the antigen-binding portion of an antibody and a non-antibody protein. In some embodiments, the antibody is selected from the group including scFv, Fab, VHH, single-domain antibody, etc. In some embodiments, the antibody is scFv. In some embodiments, the antibody is Fab. In some embodiments, the antibody is VHH. In some embodiments, the antibody is a single-domain antibody.

[0267] In some embodiments, the viral particles provided herein are pseudotyped viral particles. In some embodiments, viral particles are pseudotyped using viral glycoproteins of viruses from the Paramyxoviridae family. In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of the morbillivirus genus (such as Measlesvirus). In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Measlesvirus. In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Henipavirus (such as Nipah virus, Cedar virus, or Hendra virus). In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Nipah virus. In some embodiments, pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Nipah virus. In some embodiments, peptides or antibodies as provided herein are linked to envelope glycoproteins G or H of viruses from the Paramyxoviridae family via a linker. In some implementations, the viruses of the Paramyxoviridae family are those of the Measlesvirus genus, such as measles virus. In other implementations, the viruses of the Paramyxoviridae family are Hennipa viruses, such as Nipah virus, Cedar virus, or Hendra virus.

[0268] As provided herein, viruses can be pseudotyped using the VSV-G protein (wild type or a mutant thereof). Without being bound by any particular theory, a mutant VSV-G protein containing a mutation at position 182, which can be used to pseudotype viruses (e.g., lentiviruses) when the virus contains a target region, can be used to pseudotype the virus and transduce cells. This mutation inhibits or reduces the affinity of VSV-G for its natural co-receptor LDL-R. In some embodiments, the provided mutant VSV-G protein can be used to transduce target cells and deliver heterologous molecules to target cells.

[0269] In some embodiments, a VSV-G protein comprising a mutation at position 198 compared to SEQ ID NO: 1 or a mutation at position 182 compared to SEQ ID NO: 2 is provided. SEQ ID NO: 1 is the full-length protein, and SEQ ID NO: 2 is the extracellular domain of the VSV-G protein. The 16-mer signal peptide of MKCLLYLAFLFIGVNC (SEQ ID NO: 65) as shown at the N-terminus of SEQ ID NO: 1 is cleaved, leaving the protein of SEQ ID NO: 2. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 2, it should be understood that the mutation also occurs in the context of SEQ ID NO: 1 containing the leader sequence, and will therefore be a position number 16 positions higher than that described in SEQ ID NO: 2. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the I182D mutation compared to SEQ ID NO: 2. In some implementations, the mutation is the I182E mutation compared to SEQ ID NO: 2.

[0270] In some embodiments, a VSV-G protein comprising a mutation at position 198 compared to SEQ ID NO: 10 or a mutation at position 182 compared to SEQ ID NO: 11 is provided. SEQ ID NO: 10 is the full-length protein, and SEQ ID NO: 11 is the extracellular domain of the VSV-G protein. The 16-mer signal peptide of MLSYLIFALVVSPILG (SEQ ID NO: 66) as shown at the N-terminus of SEQ ID NO: 10 is cleaved, leaving the protein of SEQ ID NO: 11. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 11, it should be understood that the mutation also occurs in the context of SEQ ID NO: 10 containing the leader sequence, and will therefore be a position number 16 positions higher than that described in SEQ ID NO: 11. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the T182D mutation compared to SEQ ID NO: 11. In some implementations, the mutation is the T182E mutation compared to SEQ ID NO: 11.

[0271] In some embodiments, a VSV-G protein comprising a mutation at position 198 compared to SEQ ID NO: 12 or a mutation at position 182 compared to SEQ ID NO: 13 is provided. SEQ ID NO: 12 is the full-length protein, and SEQ ID NO: 13 is the extracellular domain of the VSV-G protein. The 16-mer signal peptide MLRLFLFCFLALGAHS (SEQ ID NO: 67) shown at the N-terminus of SEQ ID NO: 12 is cleaved, leaving the protein of SEQ ID NO: 13. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 13, it should be understood that the mutation also occurs in the context of SEQ ID NO: 12 containing the leader sequence, and will therefore be a position number 16 positions higher than that described in SEQ ID NO: 13. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the A182D mutation compared to SEQ ID NO: 13. In some implementations, the mutation is the A182E mutation compared to SEQ ID NO: 13.

[0272] In some embodiments, a VSV-G protein comprising a mutation at position 203 compared to SEQ ID NO: 14 or a mutation at position 182 compared to SEQ ID NO: 15 is provided. SEQ ID NO: 14 is the full-length protein, and SEQ ID NO: 15 is the extracellular domain of the VSV-G protein. The 21-mer signal peptide of MKMKMVIAGLILCIGILPAIG (SEQ ID NO: 68) as shown at the N-terminus of SEQ ID NO: 14 is cleaved, leaving the protein of SEQ ID NO: 15. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 15, it should be understood that the mutation also occurs in the context of SEQ ID NO: 14 containing the leader sequence, and will therefore be a position number 21 more than the position described in SEQ ID NO: 15. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the V182D mutation compared to SEQ ID NO: 15. In some implementations, the mutation is the V182E mutation compared to SEQ ID NO: 15.

[0273] In some embodiments, a VSV-G protein comprising a mutation at position 199 compared to SEQ ID NO: 16 or a mutation at position 182 compared to SEQ ID NO: 17 is provided. SEQ ID NO: 16 is the full-length protein, and SEQ ID NO: 17 is the extracellular domain of the VSV-G protein. The 17-mer signal peptide of MTPAFILCMLLAGSSWA (SEQ ID NO: 69), as shown at the N-terminus of SEQ ID NO: 16, is cleaved, leaving the protein of SEQ ID NO: 17. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 17, it should be understood that the mutation also occurs in the context of SEQ ID NO: 16 containing the leader sequence, and will therefore be a position number 17 more than the position described in SEQ ID NO: 17. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the V182D mutation compared to SEQ ID NO: 17. In some implementations, the mutation is the V182E mutation compared to SEQ ID NO: 17.

[0274] In some embodiments, a VSV-G protein comprising a mutation at position 199 compared to SEQ ID NO: 18 or a mutation at position 182 compared to SEQ ID NO: 19 is provided. SEQ ID NO: 18 is the full-length protein, and SEQ ID NO: 19 is the extracellular domain of the VSV-G protein. The 17-mer signal peptide MNFLLLTFIVLPLCSHA (SEQ ID NO: 70) shown at the N-terminus of SEQ ID NO: 18 is cleaved, leaving the protein of SEQ ID NO: 19. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 19, it should be understood that the mutation also occurs in the context of SEQ ID NO: 18 containing the leader sequence, and will therefore be a position number 17 positions higher than that described in SEQ ID NO: 19. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the V182D mutation compared to SEQ ID NO: 19. In some implementations, the mutation is the V182E mutation compared to SEQ ID NO: 19.

[0275] In some embodiments, a VSV-G protein comprising a mutation at position 199 compared to SEQ ID NO: 20 or a mutation at position 182 compared to SEQ ID NO: 21 is provided. SEQ ID NO: 20 is the full-length protein, and SEQ ID NO: 21 is the extracellular domain of the VSV-G protein. The 17-mer signal peptide of MLVLYLLLSLLALGAQC (SEQ ID NO: 71), as shown at the N-terminus of SEQ ID NO: 20, is cleaved, leaving the protein of SEQ ID NO: 21. Therefore, although the mutation may be mentioned in the context of SEQ ID NO: 21, it should be understood that the mutation also occurs in the context of SEQ ID NO: 20 containing the leader sequence, and will therefore be a position number 17 positions higher than that described in SEQ ID NO: 21. In some embodiments, the mutation inhibits or reduces the binding of the VSV-G protein to the LDL receptor (LDL-R). In some embodiments, the mutation is the I182D mutation compared to SEQ ID NO: 21. In some implementations, the mutation is the I182E mutation compared to SEQ ID NO: 21.

[0276] As used herein, when a peptide is considered to have a mutation compared to a reference sequence, this comparison is based on alignment software such as BlastP, ClustalW, or ClustalOmega using default parameters. For example, position 182 can be found in SEQ ID NO: 2 and is also compared with other strains as shown in Figure 3. Figure 3 illustrates the clustal alignment of the wild-type sequences of the extracellular domain of the VSV-G protein from various strains. The bolded and underlined residues are the residues aligned with position 182 of SEQ ID NO: 2 from various strains. SEQ ID NO: 2 refers to the extracellular domain of the VSV-G protein of the Indiana strain. SEQ ID NO: 11 refers to the extracellular domain of the VSV-G protein of the New Jersey strain. SEQ ID NO: 13 refers to the extracellular domain of the VSV-G protein of the Malabar strain. SEQ ID NO: 15 refers to the extracellular domain of the VSV-G protein of the Carajás strain. SEQ ID NO: 17 refers to the extracellular domain of the VSV-G protein of the Aragoa strain. SEQ ID NO: 19 refers to the extracellular domain of the VSV-G protein of the Kocal strain. SEQ ID NO: 21 refers to the extracellular domain of the VSV-G protein of the Morton strain. Therefore, residues aligned to residue 182 of SEQ ID NO: 2 can also be mutated as described herein.

[0277] In some embodiments, the mutation at position 182 compared to SEQ ID NO: 2 is not alanine. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 2 is not valine.

[0278] In some embodiments, the mutation at position 182 compared to SEQ ID NO: 2 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 11 is T182S, T182H, T182Q, or T182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 13 is A182S, A182H, A182T, A182Q, or A182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 15 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 17 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 19 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to SEQ ID NO: 21 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 is not a hydrophobic residue. In some embodiments, the mutation at position 182 is a charged residue. In some embodiments, the mutation at position 182 is a negatively charged residue.

[0279] Although the mutation may be described with reference to SEQ ID NO: 1 or SEQ ID NO: 2 (which is the VSV-G protein from the Indiana strain), the mutation can also be used in other strains of the VSV-G protein. For example, the mutation can be made in the New Jersey strain of VSV-G, the Malaba strain of VSV-G, the Carajás strain of VSV-G, the Alagoa strain of VSV-G, the Cocal strain of VSV-G, or the Morton strain of VSV-G. In some embodiments, the sequence of each strain is as provided herein. Examples of these strains can be found, for example, in U.S. Patent Application Publication No. 20200216502, which is incorporated herein by reference. For example, the wild-type full-length or extracellular domains of the New Jersey strain of VSV-G are SEQ ID NO:10 and SEQ ID NO:11, respectively; the wild-type full-length or extracellular domains of the Malaba strain of VSV-G are SEQ ID NO:12 and SEQ ID NO:13, respectively; the wild-type full-length or extracellular domains of the Carajás strain of VSV-G are SEQ ID NO:14 and SEQ ID NO:15, respectively; the wild-type full-length or extracellular domains of the Alagoa strain of VSV-G are SEQ ID NO:16 and SEQ ID NO:17, respectively; the wild-type full-length or extracellular domains of the Cocal strain of VSV-G are SEQ ID NO:18 and SEQ ID NO:19, respectively; or the wild-type full-length or extracellular domains of the Morton strain of VSV-G are SEQ ID NO:20 and SEQ ID NO:21, respectively.

[0280] The VSV-G protein containing the mutation at position 182 compared to SEQ ID NO: 2 may also contain other mutations, such as those described in U.S. Patent Application Publication No. 20200216502, the entire contents of which are incorporated herein by reference. For example, the VSV-G protein may contain mutations at positions corresponding to positions 8, 47, 209, and / or 354 of SEQ ID NO: 2.

[0281] In some embodiments, the substitution at position 8 is made by any amino acid other than Y that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 209 is made by any amino acid other than H that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 47 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 354 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2.

[0282] In some embodiments, the replacement is located at position 47 or at position 354, or at both positions 47 and 354, and is replaced by A, G, F, or Q. In some embodiments, the replacement is A or Q.

[0283] In some implementations, the substitution at position 8 is alanine, i.e., H8A.

[0284] In some implementations, the substitution at position 47 is Q or N, i.e., K47Q or K47N.

[0285] In some embodiments, the protein contains a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0286] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 2 contains a mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 2 (or SEQ ID NO: 1 if a full-length protein is used). In some embodiments, the polypeptide contains an I182D or I182E mutation. In some embodiments, the VSV-G protein contains an I182S, I182H, I182T, I182Q, or I182N mutation.

[0287] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 11 contains a mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 11 (or SEQ ID NO: 10 if a full-length protein is used). In some embodiments, the polypeptide contains a T182D or T182E mutation. In some embodiments, the VSV-G protein contains a T182S, T182H, T182Q, or T182N mutation.

[0288] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 13 contains a mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 13 (or SEQ ID NO: 12 if a full-length protein is used). In some embodiments, the polypeptide contains an A182D or A182E mutation. In some embodiments, the VSV-G protein contains an A182S, A182H, A182T, A182Q, or A182N mutation.

[0289] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 15 contains a mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 15 (or SEQ ID NO: 14 if a full-length protein is used). In some embodiments, the polypeptide contains a V182D or V182E mutation. In some embodiments, the VSV-G protein contains a V182S, V182H, V182T, V182Q, or V182N mutation.

[0290] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 17 contains a mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17 (or SEQ ID NO: 16 if a full-length protein is used). In some embodiments, the polypeptide contains a V182D or V182E mutation. In some embodiments, the VSV-G protein contains a V182S, V182H, V182T, V182Q, or V182N mutation.

[0291] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 19 contains a mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 19 (or SEQ ID NO: 18 if a full-length protein is used). In some embodiments, the polypeptide contains a V182D or V182E mutation. In some embodiments, the VSV-G protein contains a V182S, V182H, V182T, V182Q, or V182N mutation.

[0292] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 21 contains a mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 21 (or SEQ ID NO: 20 if a full-length protein is used). In some embodiments, the polypeptide contains an I182D or I182E mutation. In some embodiments, the VSV-G protein contains an I182S, I182H, I182T, I182Q, or I182N mutation.

[0293] Viral glycoprotein

[0294] The mutant VSV-G protein can be used, for example, for pseudotypening viruses (such as, but not limited to, lentiviruses). Therefore, in some embodiments, viral particles comprising the mutant VSV-G protein as provided herein are provided. In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 198 compared to SEQ ID NO: 1. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 2 contains the mutation at position 182 and is at least, or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 2 (or SEQ ID NO: 1 if a full-length protein is used). In some embodiments, the peptide comprises an I182D or I182E mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises an I182S, I182H, I182T, I182Q, or I182N mutation.

[0295] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 198 compared to SEQ ID NO: 10. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 11 contains the mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 11 (or SEQ ID NO: 10 if the full-length protein is used). In some embodiments, the polypeptide contains a T182D or T182E mutation compared to SEQ ID NO: 11. In some implementations, the VSV-G protein contains T182S, T182H, T182Q, or T182N mutations.

[0296] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 198 compared to SEQ ID NO: 12. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 13 contains the mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 13 (or SEQ ID NO: 12 if a full-length protein is used). In some embodiments, the polypeptide contains an A182D or A182E mutation compared to SEQ ID NO: 13. In some implementations, the VSV-G protein contains mutations of A182S, A182H, A182T, A182Q, or A182N.

[0297] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 203 compared to SEQ ID NO: 14. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 15 contains the mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 15. In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 15. In some implementations, the VSV-G protein contains mutations of V182S, V182H, V182T, V182Q, or V182N.

[0298] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 199 compared to SEQ ID NO: 16. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 17 contains the mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17. In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 17. In some implementations, the VSV-G protein contains mutations of V182S, V182H, V182T, V182Q, or V182N.

[0299] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 199 compared to SEQ ID NO: 18. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 19 contains the mutation at position 182 and is at least, about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 19 (or SEQ ID NO: 18 if the full-length protein is used). In some embodiments, the polypeptide contains a V182D or V182E mutation compared to SEQ ID NO: 19. In some implementations, the VSV-G protein contains mutations of V182S, V182H, V182T, V182Q, or V182N.

[0300] In some embodiments, the viral particle comprises the VSV-G protein containing a mutation at position 199 compared to SEQ ID NO: 20. In some embodiments, the protein containing a mutation at position 182 compared to SEQ ID NO: 21 contains the mutation at position 182 and is at least or about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 21 (or SEQ ID NO: 20 if the full-length protein is used). In some embodiments, the polypeptide contains an I182D or I182E mutation compared to SEQ ID NO: 21. In some implementations, the VSV-G protein contains I182S, I182H, I182T, I182Q, or I182N mutations.

[0301] In some embodiments, the VSV-G protein further comprises a mutation at the position corresponding to position 214 and / or 352 of SEQ ID NO: 2. In some embodiments, the residue corresponding to position 214 of SEQ ID NO: 2 is T214. In some embodiments, the residue corresponding to position 352 of SEQ ID NO: 2 is T352. In some embodiments, the VSV-G protein comprises a mutation corresponding to the T214N mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises a mutation corresponding to the T352A mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein comprises both T214N and T352A mutations compared to SEQ ID NO: 2. These mutations may be combined with any other mutations as provided herein. In some embodiments, the T214N and / or T352A mutations are combined with the I182E or I182D mutation. In some embodiments, the VSV-G protein comprises the amino acid sequences of SEQ ID NO: 22 and SEQ ID NO: 23, which combine I182D or I182E with the T214N and T352A mutations, respectively. These sequences are further illustrated below using leader sequences that are removed during protein processing.

[0302] VSV-G protein mutations I196D, T230N, and T368A (with leader sequence and adjusted numbering)

[0303] MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(SEQ ID NO:24)

[0304] VSV-G protein mutations: I182D, T214N, and T352A (without leader sequence)

[0305] KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(SEQ ID NO: 22)

[0306] VSV-G protein with I196E, T230N, and T368A mutations (with leader sequence and adjusted numbering)

[0307] MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(SEQ ID NO:25)

[0308] VSV-G protein with I182E, T214N and T352A mutations (without leader sequence)

[0309] KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEA VIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAAR FPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVL RTSSGYKFPLYMIGGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK (SEQ ID NO: 23)

[0310] Other strains of the VSV-G protein described herein may also contain mutations corresponding to T214N and / or T352A in SEQ ID NO: 2 and are shown in SEQ ID NO: 22 and SEQ ID NO: 23.

[0311] In some embodiments, the composition comprises mutations as described in Hwang et al., Gene Ther, Aug 2013; 20(8):807-15. (Epub, Jan 31, 2013), which is incorporated herein by reference in its entirety. For example, mutations may be at positions 230, 368, 66, and / or 162 corresponding to SEQ ID NO: 1. These positions would be 16 fewer than those in SEQ ID NO: 2 when the leader sequence is removed. In some embodiments, mutations at those positions are, for example, T230N, T368A, K66T, S162T, or any combination thereof. In some embodiments, the VSV-G protein comprises the T230N and T368A mutations. In some embodiments, the VSV-G polypeptide comprises K66T, S162T, T230N, and T368A. These positions are those corresponding to the positions in the full-length protein (SEQ ID NO: 1). In some embodiments, the VSV-G protein comprises the T230N mutation, the T368A mutation, the K66T mutation, the S162T mutation, or any combination thereof. In some embodiments, in addition to the mutation corresponding to position 182 of SEQ ID NO: 2, the VSV-G protein also comprises one or more mutations, such as those described in U.S. Patent Application Publication No. 20200216502, the entire contents of which are incorporated herein by reference. For example, the VSV-G protein may also comprise mutations at positions corresponding to positions 8, 47, 209, and / or 354 of SEQ ID NO: 2.

[0312] In some embodiments, the substitution at position 8 is made by any amino acid other than Y that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 209 is made by any amino acid other than H that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 47 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution at position 354 is made by any amino acid other than K or R that is different from the amino acid indicated at that position in sequence SEQ ID NO: 2. In some embodiments, the substitution is located at position 47 or position 354, or at both positions 47 and 354, and is substituted by A, G, F, or Q. In some embodiments, the substitution is A or Q. In some embodiments, the substitution at position 8 is alanine, i.e., H8A. In some embodiments, the substitution at position 47 is Q or N, i.e., K47Q or K47N. In some embodiments, the protein contains a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0313] In addition, in some implementations, other viral structural proteins can be used to pseudotype the virus instead of the VSV-G protein or its mutants.

[0314] For example, viral particles can be pseudotyped using the carp viremia virus G (SVCV-G) protein, and cells can be transduced when the virus contains a targeting portion. In some embodiments, the provided carp viremia virus G protein can be used to transduce target cells and deliver heterologous molecules to target cells. In some embodiments, a carp viremia virus G protein comprising SEQ ID NO: 52 is provided. SEQ ID NO: 52 is the full-length protein, and SEQ ID NO: 53 is the extracellular domain of the carp viremia virus G protein with the N-terminal signal peptide removed. Therefore, in some embodiments, the protein comprises the amino acid sequence of SEQ ID NO: 53. The carp viremia virus G protein can be used, for example, to pseudotype viruses (such as, but not limited to, lentiviruses). Therefore, in some embodiments, viral particles comprising the carp viremia virus G protein as provided herein are provided. In some embodiments, the viral particle contains a carp spring viremia virus G protein containing at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the same sequence as SEQ ID NO: 52 or SEQ ID NO: 53.

[0315] The sequence of carp spring viremia virus-G (SEQ ID NO: 52 - with leader sequence)

[0316] MSIISYIAFLLLIDSNLGIPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDLGFPPQSCGWASVTTVSNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWVEKTAGTLTTIHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSLNGTDPRTNASRVFWDDWELDGNIYQGFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENGNPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSKRPATESHEMRSLV

[0317] Sequence of carp spring viremia virus-G (SEQ ID NO: 53 - without leader sequence) :

[0318] IPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDLGFPPQSCGWASVTTVSNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWVEKTAGTLTTIHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSLNGTDPRTNASRVFWDDWELDGNIYQGFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENGNPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSKRPATESHEMRSLV

[0319] Targeted portion

[0320] In some embodiments, the viral particle includes a targeting portion having the formula T-S1, where T is a target-binding domain and S1 is a stem portion. The targeting portion can be used to target viral particles containing mutant VSV-G or SVCV-G proteins to cells expressing a target to which the targeting portion binds. In some embodiments, the target-binding domain is an antibody, scFv antibody, antigen-binding domain, an ankyrin repeat sequence (e.g., DARPIN), a VHH domain antibody, a nanobody, a single-domain antibody, an FN3 domain, or any combination thereof. The target-binding domain can be attached to the viral surface via a variant Fc protein (e.g., L1-Fc-L2-X1) as provided herein or via a flexible polypeptide (e.g., L3-X1) as provided herein. In some embodiments, the targeting portion is attached (fused to or linked) to the envelope glycoprotein G or H of a virus of the Paramyxoviridae family, such as a measlesvirus, or a Hennipa virus, such as Nipah virus, Cedar virus, or Hendra virus. In some embodiments, the targeting portion may attach (fuse or link) to a glycoprotein of a virus belonging to the Rhabdoviridae family, such as vesicular stomatitis virus (VSV), vesicular stomatitis virus (VSV), vesicular stomatitis virus (VSV), vesicular stomatitis virus (VSV), vesicular stomatitis virus (VSV), parainfluenza virus, fall armyworm rhabdovirus isolate Sf G, Drosophila sigmavirus 10A, Wuhan insect virus 7, perch virus, or carp spring viremia virus. In some embodiments, the VSV protein is a mutant protein, such as those described herein. In some embodiments, the targeting portion attaches to a glycoprotein of a virus belonging to the Filoviridae family (such as Ebola virus) or a virus belonging to the Arenaviridae family (such as Machupo virus).

[0321] In some implementations, the target-binding domain is scFv. In some implementations, the target-binding domain is a single-domain antibody. In some implementations, the target-binding domain is VHH.

[0322] In some implementations, the targeting portion binds to the following: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR α, TCR β, TCR γ, TCR δ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3; glycosylated CD43 epitopes expressed on acute leukemia or lymphoma but not on hematopoietic progenitor cells; glycosylated CD43 epitopes expressed on non-hematopoietic cancers; A kinase anchoring protein 4 (AKAP-4); adrenaline receptor β3 (ADRB3); AFP; anaplastic lymphoma kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie 2) Anti-desmosome core protein 1 (Dsg1) autoantibody; anti-desmosome core protein 3 (Dsg3) autoantibody; B7H3 (CD276); biotin; bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; cancer / testis antigen 1 (NY-ESO-1); cancer / testis antigen 2 (LAGE-la); carbonic anhydrase IX (CA1X); carcinoembryonic antigen (CEA); CCCTC binding factor (zinc finger protein)-like (BORIS or imprinted site regulator sibling factor); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32 (FCG) R2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGLll; CD200R; CD276 / B7H3; CD300 molecular-like family member f (CD300LF); CDH1-CD324; CDH6; CDH17; CDH19; Chromosome X open reading frame 61 (CXORF61); Sealing protein 6 (CLDN6); Sealing protein 18.2 (CLD18A2 or CLDN18A.2); CMV pp65; C-MYC epitope tag; Cripto; CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24); CSF2RA (GM-CSFR-α); C-type lectin domain family 12 member A (CLEC12A); C-type lectin-like molecule-1 (CLL-1 or CLECL1); Cyclin B1; Cytochrome P450 IB 1 (CYP1B 1); DLL3; EBV-EBNA3c; EGF-bke module 2 containing mucin-like hormone receptor-like receptor (EMR2); Elongation factor 2 mutant (ELF2M); Hepatin B2; Hepatin A receptor 2 (EphA2)Epidermal growth factor receptor (EGFR); Epidermal growth factor receptor variant III (EGFRviii); Epithelial cell adhesion molecule (EPCAM); ERG; ETS translocation variant gene 6 located on chromosome 12p (ETV6-AML); Fc fragment of IgA receptor (FCAR or CD89); Fc receptor-like 5 (FCRL5); Fibroblast activation protein α (FAP); FITC; Fms-like tyrosine kinase 3 (FLT3); Folate receptor α (FRa or FR1); Folate receptor β (FRb); Follicle-stimulating hormone receptor (FSHR); Fos-associated antigen 1; Fucosyl-GM1; G protein-coupled receptor class C5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; Ganglioside G2 (GD2); Ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(l-4)bDGlcp(ll)Cer); Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l- 4) bDGlcp(ll)Cer); GD3; GFRα4; Glycoprotein 100 (gplOO); Phosphatidylinositol proteoglycan-3 (GPC3); Gonadotropin receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylate cyclase C (GCC); Mutant heat shock protein 70-2 (mut hsp70-2); Hepatitis A virus cell receptor 1 (HAVCR1); GloboH glycosylceramide hexasaccharide moiety (GloboH); High molecular weight melanoma-associated antigen (HMWMAA); HIV1 envelope glycoprotein; HLA; HLA-DOA; HLA-A; HLA-A2; HLA-B; HLA-C; HLA-DM; HLA-DOB; HLA-DP; HLA-DQ; HLA-DR; HLA-G; HTLV1-Tax; Human papillomavirus E6 (HPV) E6); Human papillomavirus E7 (HPV E7); Human telomerase reverse transcriptase (hTERT); IgE; IL13Ra2; IL11Ra; Immunoglobulin λ-like polypeptide 1 (IGLL1); Influenza A hemagglutinin (HA); Insulin-like growth factor 1 receptor (IGF-I receptor); Interleukin 11 receptor α (IL-11Ra); Interleukin 13 receptor subunit α-2 (IL-13Ra2 or CD213A2); Intestinal carboxylesterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; pod protein; Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); Luteinizing hormone receptor (LHR); Lewis (Y) antigen; Lews Ag; Livl; Locus K9 (LY6K); Low-conductivity chloride channel;Lymphocyte antigen 6 complex; Lymphocyte antigen 75 (LY75); Lymphocyte-specific protein tyrosine kinase (LCK); Breast differentiation antigen (NY-BR-1); T-cell recognized melanoma antigen 1 (MelanA or MARTI); Melanoma-associated antigen 1 (MAGE-A1); Melanoma cancer testis antigen-1 (MAD-CT-1); Melanoma cancer testis antigen-2 (MAD-CT-2); Melanoma cell apoptosis inhibitor (ML-IAP); Mesothelin; MPL; Cell surface-associated mucin 1 (MUC1); N-acetylglucosamine transferase V (NA17); Ligin-4; Neural cell adhesion molecule (NCAM); NKG2D; NYBR1; O-acetyl-GD2 ganglioside (OAcGD2); olfactory receptor 51E2 (OR51E2); oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia virus oncogene homolog 1 (Abl) (bcr-abl); P53 mutant; pairing box protein Pax-3 (PAX3); pairing box protein Pax-5 (PAX5); pan-connector protein 3 (PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor β (PDGFR-β); polysialic acid; proapocrine-binding protein sp32 (OY-TES1); prostate enzyme; prostate cancer tumor antigen-1 (PCT) A-1 or galactohemagglutinin 8); prostate stem cell antigen (PSCA); prostate-specific membrane antigen (PSMA); prostatic acid phosphatase (PAP); Prostein; serine protease 21 (Testisin or PRSS21); proteasome macropain subunit β9 (LMP2); PTK7; Ras G12V; Ras homolog family member C (RhoC); Rat sarcoma (Ras) mutant; Receptor to advanced glycation end products (RAGE-1); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Receptor tyrosine protein kinase ERBB2 or Her-22 / neu; Renal pervasive protein 1 (RU1); Renal pervasive protein 2 (RU2); Sarcoma translocation breakpoint; Serine 2 (TMPRSS2) ETS fusion gene; Sialoyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialic acid Lewis antigen); Sperm protein 17 (SPA17); T cell recognized squamous cell carcinoma antigen 3 (SART3); Stage-specific embryonic antigen-4 (SSEA-4); STEAP1; Survival protein; Synovial sarcoma X breakpoint 2 (SSX2); TCR γ-substituted reading frame protein (TARP); TCR-β1 chain; TCR-β2 chain; TCR-δ chain; TCR-γ chain; TCRγ-δ; telomerase; TGFβR2; antigen recognized by TNT antibody;Thyroid-stimulating hormone receptor (TSHR); Timl- / HVCR1; Tissue factor 1 (TF1); Tn ag; Tn antigen ((Tn Ag) or (GalNAca-Ser / Thr)); TNF receptor family member B cell maturation (BCMA); transglutaminase 5 (TGS5); transmembrane protease; TROP2; tumor endothelial marker 1 (TEM1 / CD248); tumor endothelial marker 7 associated (TEM7R); tumor protein p53 (p53); tumor-associated glycoprotein 72 (TAG72); tyrosinase; tyrosinase-associated protein 2 (TRP-2); urolytic protein 2 (UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein (WT1); or X antigen family member 1A (XAGE1). In some embodiments, the targeting portion binds to CD7. In some embodiments, the targeting portion binds to CD8.

[0323] In some embodiments, the targeting portion binds to a target present on cells, such as immune cells. In some embodiments, the cells are immune cells, such as, but not limited to, T cells, B cells; NK cells, dendritic cells, neutrophils, macrophages, cancer cells; or, for example, CD3+ T cells; CD4+ T cells, CD7+ T cells, CD8+ T cells; CD19+ B cells, CD19+ cancer cells; CD20+ B cells, CD20+ cancer cells; CD30+ lung epithelial cells; CD34+ hematopoietic stem cells; CD105+ endothelial cells; CD105+ hematopoietic stem cells; CD117+ hematopoietic stem cells; CD133+ cancer cells; EpCAM+ cancer cells; GluA2+ neurons; GluA4+ neurons; hematopoietic stem cells; hepatocytes; Her2 / Neu+ cancer cells; NKG2D+ natural killer cells; SLC1A3+ astrocytes; SLC7A10+ adipocytes. In some embodiments, the cells are T cells. In some embodiments, the cells are B cells. In some implementations, the cells are CD7+ T cells and / or CD8+ T cells.

[0324] CD7-binding peptide

[0325] In some implementations, the target moiety (e.g., a peptide) binds to CD7.

[0326] In some embodiments, the CD7-binding peptide is an antibody that binds to non-human primate CD7. In some embodiments, the CD7-binding peptide is an antibody that binds to human CD7. The sequence of human CD7 (UniProtKB P09564) is as follows (SEQ ID NO: 29):

[0327] MAGPPRLLLLPLLLALARGLPGALAAQEVQQSPHCTTVPVGASVNITCSTSGGLRGIYLRQLGPQPQDIIYYEDGVVPTTDRRFRGRIDFSGSQDNLTITMHRLQLSDTGTYTCQAITEVNV YGSGTLVLVTEEQSQGWHRCSDAPPRASALPAPPTGSALPDPQTASALPDPPAASALPAALAVISFLLGLGVACVLARTQIKKLCSWRDKNSAACVVYEDMSHSRCNTLSSPNQYQ (SEQ ID NO: 29).

[0328] In some embodiments, the peptide-bound CD7 is expressed on the cell surface. In some embodiments, the cell is an immune cell. In some embodiments, the immune cell is a CD7+ T cell, CD4+ T cell, CD8+ T cell, NK cell, α-β T cell, γ-δ T cell, lymphoprogenitor cell, hematopoietic stem cell, bone marrow cell, monocyte, macrophage, central memory T cell, effector memory T cell, stem cell-like memory T cell, naive T cell, activated T cell, regulatory T cell (TReg), terminally differentiated effector memory T cell (TEMRA), resident memory T cell (TRM), or T cell CD8+CCR7+.

[0329] In some embodiments, the antibody comprises an Fc region. The Fc region may be linked to either the heavy or light chain of the antibody. In some embodiments, the Fc region is an IgG Fc. In some embodiments, the IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, the IgG fc is an IgG1 Fc. In some embodiments, the antibody comprises a constant Fc region as shown herein, such as SEQ ID NO: 26, 27, or 28, or a mutant thereof as provided herein.

[0330] In some embodiments, this document provides a peptide (e.g., a CD7-binding peptide). In some embodiments, this document provides an antibody (e.g., an anti-CD7 antibody). In some embodiments, the antibody is a recombinant antibody that binds to CD7. In some embodiments, the CD7 protein is the human CD7 protein. In some embodiments, the CD7 protein is a non-human CD7 protein (e.g., mouse, rat, pig, dog, non-human primate). As used herein, the term "recombinant antibody" refers to an antibody that is not naturally occurring. In some embodiments, the term "recombinant antibody" refers to an antibody that has not been isolated from a human subject.

[0331] In some embodiments, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antibody fragment comprises a peptide selected from the table below, which describes the CDR based on Kabat numbering.

[0332]

[0333] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to non-human primate CD7. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to human CD7. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 33-35. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 33. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 34. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 35. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having a sequence selected from SEQ ID NO: 30-32. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 30. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 31. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 32. The CDRs mentioned throughout the embodiments of this specification are interchangeable with CDRs characterized in different formats, such as Chothia and IMGT.

[0334] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain variable region having LCDR1, LCDR2, and LCDR3, wherein LCDR1 has the sequence of SEQ ID NO: 33, LCDR2 has the sequence of SEQ ID NO: 34, and LCDR3 has the sequence of SEQ ID NO: 35.

[0335] In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain variable region having HCDR1, HCDR2, and HCDR3, wherein HCDR1 has the sequence of SEQ ID NO: 30, HCDR2 has the sequence of SEQ ID NO: 31, and HCDR3 has the sequence of SEQ ID NO: 32.

[0336] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises: (i) a light chain having any of the aforementioned combinations of LCDR1, LCDR2, and LCDR3 sequences; and (ii) a heavy chain having any of the aforementioned combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0337] Different CDR motifs can be combined in any combination (including those not described in the table above). For example, the following embodiments are provided as non-limiting examples of such combinations.

[0338] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 33; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 34; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 35; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 30; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 31; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 32; or a variation thereof.

[0339] Although the preceding paragraphs may refer to CDRs according to the Kabat system, equivalent CDR sequences from the names IMGT and CHOTHIA can be used.

[0340] In some embodiments, the light chain variable region CDR1 is replaced by any of the other light chain CDR1 sequences. In some embodiments, the light chain variable region CDR2 is replaced by any of the other light chain CDR2 sequences. In some embodiments, the light chain variable region CDR3 is replaced by any of the other light chain CDR3 sequences. In some embodiments, the heavy chain variable region CDR1 is replaced by any of the other heavy chain CDR1 sequences. In some embodiments, the heavy chain variable region CDR2 is replaced by any of the other heavy chain CDR2 sequences. In some embodiments, the heavy chain variable region CDR3 is replaced by any of the other heavy chain CDR3 sequences.

[0341] In some embodiments, the polypeptide comprises a heavy chain variable region peptide or a variant thereof having one of the following sequences:

[0342]

[0343] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences or a variant thereof:

[0344]

[0345] In some embodiments, the peptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 36. H Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 37. L Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof contains V. H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36 or a variant thereof; and V L The peptide comprises the sequence of SEQ ID NO: 37 or a variant thereof. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36 or a variant thereof; and V L The peptide comprises the sequence of SEQ ID NO: 37 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to non-human primate CD7. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36 or a variant thereof; and V L The peptide comprises the sequence of SEQ ID NO: 37 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to human CD7. In some embodiments, V H The peptide contains the sequence of SEQ ID NO: 36; and V L The peptide contains the sequence of SEQ ID NO: 37.

[0346] V H and V L The sequence can be of any form, including but not limited to the scFv form, where V H and V LThe region connects to the peptide linker. Examples of peptide linkers that can be used to connect the various peptides described herein include, but are not limited to: (GGGGS) n (SEQ ID NO: 55), where each n is independently 1-5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, the variable region is not linked to the peptide linker. In some embodiments, the polypeptide comprises SEQ ID NO: 36 and SEQ ID NO: 37.

[0347] As provided in this article, peptides, antibodies, or their antigen-binding fragments can be sequence variants.

[0348] The sequences of peptides or antibodies can be modified to generate human IgG antibodies. Transformations of the sequences provided herein can be modified to generate other types of antibodies. CDRs can also be linked with other antibodies, proteins, or molecules to generate antibody fragments that bind to CD7.

[0349] In some embodiments, the peptides or antibodies provided herein are targeting portions on the surface of engineered viral particles. In some embodiments, the targeting portion allows binding to target cells. In some embodiments, the targeting portion is a CD7 binding portion, such as the peptides or antibodies provided herein. In some embodiments, the target-binding domain (“T”) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 38.

[0350] DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGG GSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS (SEQ ID NO: 38)

[0351] Alternatively, it may be substantially similar to SEQ ID NO: 38, or an active fragment of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises at least 90% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises at least 95% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises at least 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) comprises the sequence shown in SEQ ID NO: 38. In some embodiments, the target-binding domain (“T”) shown in SEQ ID NO: 38 is an antibody or an antigen-binding fragment thereof. In some embodiments, the target-binding domain (“T”) is an anti-CD7 antibody.

[0352] In some embodiments, the peptide or antibody, as provided herein, is a targeting portion on the surface of an engineered viral particle. In some embodiments, the engineered viral particle is a pseudotyped virus-like particle. In some embodiments, the targeting portion allows binding to target cells. In some embodiments, the targeting portion is a CD7 binding portion, such as the peptide or antibody provided herein. In some embodiments, the target-binding domain (“T”) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) comprises at least 90% of the sequence of SEQ ID NO: 39.

[0353] QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGS GGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (SEQ ID NO: 39)

[0354] Alternatively, it may be substantially similar to SEQ ID NO: 39, or an active fragment of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) comprises a sequence that is at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) comprises a sequence that is at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain (“T”) as shown in SEQ ID NO: 39 is an antibody or an antigen-binding fragment thereof. In some embodiments, the target-binding domain (“T”) is an anti-CD7 antibody. In some embodiments, the anti-CD7 antibody binds to non-human primate CD7. In some embodiments, the anti-CD7 antibody binds to human CD7.

[0355] CD8-binding peptide

[0356] In some implementations, the targeting portion (e.g., a peptide) can bind to CD8.

[0357] In some embodiments, the peptide binds to CD8. In some embodiments, the peptide binds to CD8-α. In some embodiments, the peptide binds to CD8-β. In some embodiments, the peptide binds to a CD8 heterodimer. In some embodiments, the CD8 heterodimer comprises CD8-α and CD8-β subunits. In some embodiments, the peptide binds to a CD8-α homodimer. In some embodiments, the peptide binding to CD8 is an antibody binding to non-human primate CD8. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8-α. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8-β. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8-α homodimer. In some embodiments, the antibody binding to non-human primate CD8 is an antibody binding to non-human primate CD8 heterodimer. In some embodiments, the CD8-binding peptide is an antibody that binds to human CD8. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-α. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-β. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-α homodimer. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8 heterodimer. The sequence of human CD8-α (UniProtKB Q8TAW8) is as follows (SEQ ID NO: 40):

[0358] MALPPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGCYFCSALS NSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV (SEQ ID NO: 40).

[0359] The sequence of human CD8-β (UniProtKB Q8TD28) is as follows (SEQ ID NO: 41):

[0360] MRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKISLSNMRIYWLRQRQAPSSDSHHEFLALWDSAKGTIHGEEVEQEKIAVFRDASRFILNLTSVK PEDSGIYFCMIVGSPELTFGKGTQLSVVDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCSPITLGLLVAGVLVLLVSLGVAIHLCCRRRRARLRFMKQLYK (SEQ ID NO:41).

[0361] In some embodiments, the peptide-bound CD8 is expressed on the cell surface. In some embodiments, the cell is an immune cell. In some embodiments, the immune cell is a CD7+ T cell, CD4+ T cell, CD8+ T cell, NK cell, α-β T cell, γ-δ T cell, lymphoprogenitor cell, hematopoietic stem cell, bone marrow cell, monocyte, macrophage, central memory T cell, effector memory T cell, stem cell-like memory T cell, naive T cell, activated T cell, regulatory T cell (TReg), terminally differentiated effector memory T cell (TEMRA), resident memory T cell (TRM), or T cell CD8+CCR7+. In some embodiments, the cell is a CD8+ T cell. In some embodiments, the cell is a CD8+ cell.

[0362] In some embodiments, the antibody comprises an Fc region. The Fc region may be linked to either the heavy or light chain of the antibody. In some embodiments, the Fc region is an IgG Fc. In some embodiments, the IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, the IgG Fc is an IgG1 Fc. In some embodiments, the antibody comprises a constant Fc region as shown herein, such as SEQ ID NO: 26, 27, or 28, or a variant thereof.

[0363] In some embodiments, this document provides a peptide (e.g., a CD8-binding peptide). In some embodiments, this document provides an antibody (e.g., an anti-CD8 antibody). In some embodiments, the antibody is a recombinant antibody that binds to CD8. In some embodiments, the CD8 protein is a human CD8 protein. In some embodiments, the CD8 protein is a non-human CD8 protein (e.g., mouse, rat, pig, dog, non-human primate). As used herein, the term "recombinant antibody" refers to an antibody that is not naturally occurring. In some embodiments, the term "recombinant antibody" refers to an antibody that has not been isolated from a human subject.

[0364] In some embodiments, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antibody fragment comprises a peptide selected from the table below, which describes the CDR based on Kabat numbering.

[0365]

[0366] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to non-human primate CD8. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain or light chain CDR as provided in the table above and binds to human CD8. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 45-47. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 45. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 46. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a light chain CDR having a sequence selected from SEQ ID NO: 47. In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having a sequence selected from SEQ ID NO: 42-44. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 42. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 43. In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO: 44. The CDRs mentioned throughout the embodiments of this specification may be interchanged with CDRs characterized in different formats, such as Chothia and IMGT.

[0367] In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a light chain variable region having LCDR1, LCDR2, and LCDR3, wherein LCDR1 has the sequence of SEQ ID NO: 45, LCDR2 has the sequence of SEQ ID NO: 46, and LCDR3 has the sequence of SEQ ID NO: 47.

[0368] In some embodiments, the peptide, antibody, or antibody-binding fragment thereof comprises a heavy chain variable region having HCDR1, HCDR2, and HCDR3, wherein HCDR1 has the sequence of SEQ ID NO: 42, HCDR2 has the sequence of SEQ ID NO: 43, and HCDR3 has the sequence of SEQ ID NO: 44.

[0369] In some embodiments, the polypeptide, antibody, or antibody-binding fragment thereof comprises: (i) a light chain having any of the aforementioned combinations of LCDR1, LCDR2, and LCDR3 sequences; and (ii) a heavy chain having any of the aforementioned combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0370] Different CDR motifs can be combined in any combination (including those not described in the table above). For example, the following embodiments are provided as non-limiting examples of such combinations.

[0371] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 45; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 46; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 47; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 42; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 43; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 44; or a variation thereof.

[0372] Although the preceding paragraphs may refer to CDRs according to the Kabat system, equivalent CDR sequences from the names IMGT and CHOTHIA can be used.

[0373] In some embodiments, the light chain variable region CDR1 is replaced by any of the other light chain CDR1 sequences. In some embodiments, the light chain variable region CDR2 is replaced by any of the other light chain CDR2 sequences. In some embodiments, the light chain variable region CDR3 is replaced by any of the other light chain CDR3 sequences. In some embodiments, the heavy chain variable region CDR1 is replaced by any of the other heavy chain CDR1 sequences. In some embodiments, the heavy chain variable region CDR2 is replaced by any of the other heavy chain CDR2 sequences. In some embodiments, the heavy chain variable region CDR3 is replaced by any of the other heavy chain CDR3 sequences.

[0374] In some embodiments, the polypeptide comprises a heavy chain variable region peptide or a variant thereof having one of the following sequences:

[0375]

[0376] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences or a variant thereof:

[0377]

[0378] In some embodiments, the peptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 48. H Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V of SEQ ID NO: 49. L Peptide. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof contains V. H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 48 or a variant thereof; and V L The peptide comprises the sequence of SEQ ID NO: 49 or a variant thereof. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 48 or a variant thereof; and V L The peptide comprises the sequence of SEQ ID NO: 49 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to non-human primate CD8. In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 48 or a variant thereof; and V L The peptide comprises the sequence of SEQ ID NO: 49 or a variant thereof, and the polypeptide, antibody, or antigen-binding fragment thereof binds to human CD8. In some embodiments, V H The peptide contains the sequence of SEQ ID NO: 48; and V L The peptide contains the sequence of SEQ ID NO: 49.

[0379] V H and V L The sequence can be of any form, including but not limited to the scFv form, where V H and V LThe region is attached to a peptide linker. Examples of peptide linkers that can be used to link the various peptides provided herein include, but are not limited to: (GGGGS)n (SEQ ID NO: 55), where each n is independently 1-5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, the variable region is not attached to a peptide linker. In some embodiments, the polypeptide comprises SEQ ID NO: 48 and SEQ ID NO: 49.

[0380] As provided in this article, peptides, antibodies, or their antigen-binding fragments can be sequence variants.

[0381] The sequences of peptides or antibodies can be modified to generate human IgG antibodies. Transformations of the sequences provided herein can be modified to generate other types of antibodies. CDRs can also be linked with other antibodies, proteins, or molecules to generate antibody fragments that bind to CD8.

[0382] In some embodiments, the peptides or antibodies provided herein are targeting portions on the surface of engineered viral particles. In some embodiments, the targeting portion allows binding to target cells. In some embodiments, the target-binding domain (“T”) is a CD8 binding portion, such as the peptides or antibodies provided herein. In some embodiments, the target-binding domain (“T”) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 50.

[0383] NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGGGGSGGGGSGGGGSG GGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS (SEQ ID NO: 50)

[0384] Alternatively, it may be substantially similar to SEQ ID NO: 50, or an active fragment of SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) comprises at least 90% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) comprises at least 95% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) comprises at least 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) comprises the sequence shown in SEQ ID NO: 50. In some embodiments, the target-binding domain (“T”) as shown in SEQ ID NO: 50 is an antibody or an antigen-binding fragment thereof. In some embodiments, the targeting portion is an anti-CD8 antibody.

[0385] In some embodiments, the peptide or antibody, as provided herein, is a targeting portion on the surface of an engineered viral particle. In some embodiments, the engineered viral particle is a pseudotyped virus-like particle. In some embodiments, the targeting portion allows binding to target cells. In some embodiments, the target-binding domain (“T”) is a CD8 binding portion, such as the peptide or antibody provided herein. In some embodiments, the target-binding domain (“T”) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 51.

[0386] EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSSGGGGSGG GGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR (SEQ ID NO: 51)

[0387] Alternatively, it may be substantially similar to SEQ ID NO: 51, or an active fragment of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises at least 90% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises at least 95% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises at least 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) comprises the sequence shown in SEQ ID NO: 51. In some embodiments, the target-binding domain (“T”) shown in SEQ ID NO: 51 is an antibody or an antigen-binding fragment thereof. In some embodiments, the targeting portion is an anti-CD8 antibody. In some embodiments, the anti-CD8 antibody binds to non-human primate CD8. In some embodiments, the anti-CD8 antibody binds to human CD8.

[0388] Target portion containing Fc domain

[0389] In some implementation schemes, V H and V LThe polypeptide is linked to a stem portion S1 containing the Fc region. In some embodiments, the Fc region is as provided herein. In some embodiments, the Fc region is a variant Fc region as provided herein. Non-restrictive mutations in the Fc region are provided herein. In some embodiments, the variant Fc region contains a sequence as a variant of SEQ ID NO:26, SEQ ID NO:27, or SEQ ID NO:28 as provided herein. In some embodiments, the variant of SEQ ID NO:26 contains one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A as provided herein. In some embodiments, the variant of SEQ ID NO:27 contains one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A as provided herein. In some embodiments, the variant of SEQ ID NO: 28 comprises one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A as provided herein. As provided herein, the heavy chain may be linked to the Fc region. In some embodiments, the Fc region also comprises (e.g., linked to) a transmembrane domain. In some embodiments, the Fc region also comprising the transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising the transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. Examples of ECDs include, but are not limited to, the CD8 and / or CD28 extracellular domains as described herein. T M Examples include, but are not limited to, CD8 and / or CD28 transmembrane domains as provided herein. In some embodiments, X1 includes T M Furthermore, ECD and ICD are not present. In some implementations, X1 includes ECD and T. M And ICD does not exist. In some implementations, X1 includes T. M And ICD, but ECD is not present. In some implementations, X1 includes ECD, T MAnd ICD. In any of the following embodiments, it should be understood that ECD, ICD, or both may optionally be absent. Thus, X1 comprises CD8 and / or CD28 ECD, CD8 and / or CD28 T. M Implementations of ICDs containing Env-doped motifs are understood to include the following X1 members: i) CD8 and / or CD28 ECD, CD8 and / or CD28 T M and ICDs containing Env-doped motifs; ii) CD8 and / or CD28 T M and ICDs containing Env-doped motifs, where ECDs are absent; iii) CD8 and / or CD28 ECDs and CD8 and / or CD28 T M iv) where ICD is absent; and iv) CD8 and / or CD28 T M Where neither ECD nor ICD exists. Similarly, where X1 contains CD8 and / or CD28 T. M Implementations of ICDs containing Env-doped motifs are understood to include the following X1 members: i) CD8 and / or CD28 T M and ICDs containing Env-doped motifs; and ii) CD8 and / or CD28 T M Where ICD is absent. Similarly, where X1 contains CD8 and / or CD28 ECD and CD8 and / or CD28 T. M The implementation scheme is understood to include the following X1 members: i) CD8 and / or CD28 ECD and CD8 and / or CD28 T M ; and ii) CD8 and / or CD28 T M The ECD is not present. It should also be understood that the foregoing explanation does not address specific ECDs or Ts. M This is also correct in the implementation scheme of the ICD. For example, where X1 includes an ECD, CD8, and / or CD28 T. M The implementation scheme of ICD will be understood to include the following X1 members: i) ECD, CD8 and / or CD28 T M and ICD; ii) ECD and CD8 and / or CD28 T M iii) CD8 and / or CD28 T M and ICD, where ECD is absent; and iv) CD8 and / or CD28 T M Where ECD and ICD are absent. Similarly, where X1 contains CD8 and / or CD28 T. M The implementation scheme of ICD is understood to include the following X1 members: i) CD8 and / or CD28T Mand ICD; and ii) CD8 and / or CD28 T M Where ICD is absent. Similarly, where X1 contains ECD and CD8 and / or CD28T. M The implementation scheme is understood to include the following X1 members: i) ECD and CD8 and / or CD28 T M ; and ii) CD8 and / or CD28T M Where the ECD does not exist. Unless otherwise stated, the foregoing examples and explanations apply to any of the following implementations.

[0390] In some implementations, X1 includes CD8 and / or CD28 ECD, T M And ICD. In some implementations, X1 includes an ECD, CD8, and / or CD28 T. M and ICD. In some implementations, X1 includes CD8 and / or CD28 ECD, CD8 and / or CD28 TCD. M and ICD. In some implementations, X1 includes ECD, T M and ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, X1 contains CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, X1 comprises ECD, CD8, and / or CD28 T. M And ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, X1 comprises CD8 and / or CD28 ECD, CD8 and / or CD28 TCD. M and ICD, wherein the ICD contains env-incorporated motifs as provided herein.

[0391] In some implementation schemes, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing the Fc region as described herein. In some embodiments, the V region described herein... H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region comprising a transmembrane domain as described herein. In some embodiments, the Fc region comprising the transmembrane domain also has the formula L1-Fc-L2-X1, where L1 is a linker as described herein or is absent, Fc is a variant Fc region as described herein, L2 is a linker as described herein or is absent, and X1 is a polypeptide comprising the transmembrane domain as described herein. As provided herein, X1 may comprise a portion having the formula ECD-T M-ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the V provided herein... H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M and ICD. In some implementations, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8, and / or CD28T. M and ICD. In some implementations, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M and ICD. In some implementations, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1), which includes ECD and T. M and ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M and ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8, and / or CD28 T. M and ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, the V provided herein H and V L The polypeptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. Mand ICD, wherein the ICD contains env-incorporated motifs as provided herein. In some embodiments, the env-incorporated motifs provided herein are linked to the stem portion (S1) containing the Fc region (L1-Fc-L2-X1). H and V L The peptide anchors to the surface of viral particles (such as those described in this article), and the Fc region contains ECD, T... M and ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0392] In some implementations, V having a sequence such as SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD and T. M and ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H peptides and V having the sequence shown in SEQ ID NO:37 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes CD8 and / or CD28 ECD, T M and ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, CD8, and / or CD28 T. M and ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 36 H Peptide and V having the sequence shown in SEQ ID NO: 37 LThe peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD and T. M and ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 36. H peptides and V having the sequence shown in SEQ ID NO:37 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes CD8 and / or CD28 ECD, T M and ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 36. H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, CD8, and / or CD28 T. M and ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 36. H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M and ICD, wherein the ICD contains the env incorporation motif as provided herein. In some embodiments, the V motif having the sequence shown in SEQ ID NO: 36 is linked to the stem portion (S1) containing the Fc region (L1-Fc-L2-X1). H Peptide and V having the sequence shown in SEQ ID NO: 37 L The peptide anchors to the surface of viral particles (such as those described in this article), and this Fc region contains ECD, T... M and ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0393] In some embodiments, V comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:37. L The peptide is linked to a stem portion (S1) containing the Fc region as described herein. In some embodiments, a V-shaped portion comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L The peptide is linked to a stem portion (S1) comprising an Fc region containing a transmembrane domain as provided herein. In some embodiments, the Fc region, which also includes a transmembrane domain, has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, V comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to a compound containing ECD, T MAnd the stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to ECD containing CD8 and / or CD28, T M And the stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to a T-cell containing ECD, CD8 and / or CD28 M And the stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to ECD containing CD8 and / or CD28, CD8 and / or CD28 T M And the stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. HPeptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to a compound containing ECD, T M The stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD, wherein the ICD contains the env incorporation motif as provided herein. In some embodiments, the V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to ECD containing CD8 and / or CD28, T M The stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD, wherein the ICD contains the env incorporation motif as provided herein. In some embodiments, the V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to a T-cell containing ECD, CD8 and / or CD28 M The stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD, wherein the ICD contains the env incorporation motif as provided herein. In some embodiments, the V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. LPeptide linked to ECD containing CD8 and / or CD28, CD8 and / or CD28 T M The stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD, wherein the ICD contains the env incorporation motif as provided herein. In some embodiments, the V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 36. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37. L Peptide linked to a compound containing ECD, T M And the stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD, and anchored to the surface of viral particles (such as those provided herein). In some embodiments, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0394] In some implementations, V is provided H Peptides and Vitamins L peptides are polypeptides, in which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO:36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 37.

[0395] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V LThe peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V H Peptides and Vitamins L The peptide comprises a light chain CDR having the sequence of SEQ ID NO: 33-35; and / or a heavy chain CDR having the sequence of SEQ ID NO: 30-32. In some embodiments, the polypeptide comprises V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V H Peptides and Vitamins L The peptide comprises a light chain CDR1 having the sequence of SEQ ID NO: 33; a light chain CDR2 having the sequence of SEQ ID NO: 34; a light chain CDR3 having the sequence of SEQ ID NO: 35; and / or a heavy chain CDR1 having the sequence of SEQ ID NO: 30; a heavy chain CDR2 having the sequence of SEQ ID NO: 31; and a heavy chain CDR3 having the sequence of SEQ ID NO: 32. In some embodiments, V H or V L The CDRs in the chain are shown in the combinations provided in this article.

[0396] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V LThe peptide comprises LCDR1 having the sequence of SEQ ID NO: 33; LCDR2 having the sequence of SEQ ID NO: 34; and LCDR3 having the sequence of SEQ ID NO: 35; and V H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 30; HCDR2 having the sequence of SEQ ID NO: 31; and HCDR3 having the sequence of SEQ ID NO: 32.

[0397] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 36; and V L The peptide contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical sequence to the sequence of SEQ ID NO: 37; condition V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 33, wherein LCDR1 contains at most one conserved amino acid substitution; LCDR2 having the sequence of SEQ ID NO: 34, wherein LCDR2 contains at most one conserved amino acid substitution; and LCDR3 having the sequence of SEQ ID NO: 35, wherein LCDR3 contains at most one conserved amino acid substitution; and V H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 30, wherein HCDR1 contains at most one conserved amino acid substitution; HCDR2 having the sequence of SEQ ID NO: 31, wherein HCDR2 contains at most one conserved amino acid substitution; and HCDR3 having the sequence of SEQ ID NO: 32, wherein HCDR3 contains at most one conserved amino acid substitution.

[0398] In some implementations, the polypeptide contains V H Peptides and Vitamins L peptides, of which V H The peptide contains the sequence of SEQ ID NO: 36, and V L The peptide contains the sequence of SEQ ID NO: 37.

[0399] In some embodiments, the peptides provided herein bind to non-human primate CD7. In some embodiments, the peptides provided herein bind to human CD7.

[0400] As provided in this article, the different polypeptides (V) described herein H or V L This can be linked to a peptide linker or not to form a continuous sequence. In some embodiments, the peptide linker comprises (GGGGS). n The sequence (SEQ ID NO: 55), wherein each n is independently 1-5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. The linked peptide form can be derived from formula V. H -ZV L or V L -ZV H This indicates that Z represents a peptide linker. In some embodiments, Z is (GGGGS). n (SEQ ID NO: 55), where each n is independently 1-5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0401] In some implementations, it includes formula V L -ZV H The peptide representing the linked peptide comprises a heavy chain variable region, as shown in SEQ ID NO: 36, linked to a light chain variable region, as shown in SEQ ID NO: 37, via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, it comprises a heavy chain variable region, as shown in SEQ ID NO: 36, linked to a light chain variable region, as shown in SEQ ID NO: 37, via a peptide linker. H Connected V L The polypeptide has the sequence shown below.

[0402] DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGG GSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS (SEQ ID NO: 38).

[0403] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises at least 90% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises at least 95% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises at least 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide comprises the sequence shown in SEQ ID NO: 38. In some embodiments, the polypeptide shown in SEQ ID NO: 38 is an antibody or an antigen-binding fragment thereof. In some embodiments, the antibody is an anti-CD7 antibody. In some embodiments, the anti-CD7 antibody binds to non-human primate CD7. In some embodiments, the anti-CD7 antibody binds to human CD7.

[0404] In some implementations, it includes formula V H -ZV L The peptide representing the linked peptide comprises a light chain variable region, as shown in SEQ ID NO: 37, linked to a heavy chain variable region, as shown in SEQ ID NO: 36, via a linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, it comprises a light chain variable region, as shown in SEQ ID NO: 37, linked to a heavy chain variable region, as shown in SEQ ID NO: 36, via a peptide linker. L Connected V H The polypeptide has the sequence shown below.

[0405] QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGS GGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (SEQ ID NO: 39).

[0406] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide comprises at least 90% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide comprises at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide comprises at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide comprises the sequence shown in SEQ ID NO: 39. In some embodiments, the polypeptide shown in SEQ ID NO: 39 is an antibody or an antigen-binding fragment thereof. In some embodiments, the antibody is an anti-CD7 antibody. In some embodiments, the anti-CD7 antibody binds to non-human primate CD7. In some embodiments, the anti-CD7 antibody binds to human CD7.

[0407] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region, which further comprises transmembrane domains, such as those provided herein. In some embodiments, the Fc region comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T MIt is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T M And ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8, and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO:38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T MAnd an ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 38 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, a polypeptide comprising at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO:38 and comprising a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising ECD, T M and ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0408] In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region, which further comprises transmembrane domains, such as those provided herein. In some embodiments, the Fc region comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T M And ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8, and / or CD28 T. MAnd ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO:39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T M And ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8, and / or CD28 T. M And an ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39 and comprises a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T MAnd ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, a polypeptide comprising at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO:39 and comprising a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising ECD, T M and ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0409] In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and a stem portion (S1) containing an Fc region, the Fc region further comprising a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a portion having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T MAnd ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T M And ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 38 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, a polypeptide comprising a sequence having the sequence shown in SEQ ID NO: 38 and comprising a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising an ECD, T M and ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0410] In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and a stem portion (S1) containing an Fc region (such as those provided herein). In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and a stem portion (S1) containing an Fc region, the Fc region further comprising a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, wherein L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain as provided herein. As provided herein, X1 may comprise a polypeptide having the formula ECD-T M -ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein, or is absent, T M It is a transmembrane domain as described herein, and the ICD is an intracellular domain as described herein or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, T MAnd ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, T M And ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an env incorporation motif as provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence shown in SEQ ID NO: 39 and includes a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T M And ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, a polypeptide comprising a sequence having the sequence shown in SEQ ID NO: 39 and comprising a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) is anchored to the surface of a viral particle (such as those provided herein), the Fc region comprising an ECD, T M and ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, the peptide binds to immune cells, such as those provided herein.

[0411] In some embodiments, the polypeptide comprising formula T-S1 (where T is a target-binding domain and S1 is a stem portion) provided herein comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the amino acid sequence identical to that of SEQ ID NO: 98.

[0412] (SEQ ID NO: 98)

[0413] Alternatively, it may be substantially similar to SEQ ID NO: 98, or an active fragment of SEQ ID NO: 98. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 90% of the amino acid sequence identical to that of SEQ ID NO: 98. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 95% of the amino acid sequence identical to that of SEQ ID NO: 98. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 98% of the amino acid sequence identical to that of SEQ ID NO: 98. In some embodiments, the polypeptide comprising T-S1 provided herein contains the amino acid sequence of SEQ ID NO: 98.

[0414] In some embodiments, the polypeptide comprising formula T-S1 (where T is a target-binding domain and S1 is a stem portion) provided herein comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the amino acid sequence identical to that of SEQ ID NO: 126.

[0415] (SEQ ID NO:126)

[0416] In some embodiments, the polypeptide containing formula T-S1 provided herein contains at least 90% of the amino acid sequence identical to the sequence of SEQ ID NO: 126. In some embodiments, the polypeptide containing formula T-S1 provided herein contains at least 95% of the amino acid sequence identical to the sequence of SEQ ID NO: 126. In some embodiments, the polypeptide containing formula T-S1 provided herein contains at least 98% of the amino acid sequence identical to the sequence of SEQ ID NO: 126. In some embodiments, the polypeptide containing formula T-S1 provided herein contains the amino acid sequence of SEQ ID NO: 126.

[0417] In some embodiments, the polypeptide comprising formula T-S1 (where T is a target-binding domain and S1 is a stem portion) provided herein comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the amino acid sequence identical to the sequence of SEQ ID NO: 127.

[0418] (SEQ ID NO:127)

[0419] In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 90% of the amino acid sequence identical to the sequence of SEQ ID NO: 127. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 95% of the amino acid sequence identical to the sequence of SEQ ID NO: 127. In some embodiments, the polypeptide comprising T-S1 provided herein contains at least 98% of the amino acid sequence identical to the sequence of SEQ ID NO: 127. In some embodiments, the polypeptide comprising T-S1 provided herein contains the amino acid sequence of SEQ ID NO: 127.

[0420] In some embodiments, the polypeptide comprising formula T-S1 provided herein (where T is a target-binding domain and S1 is a stem portion) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the sequence of SEQ ID NO: 98, wherein the target-binding domain “T” of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and wherein the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises at least 90% of the amino acid sequence of SEQ ID NO: 98, wherein the target-binding domain “T” of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and wherein the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and wherein the stem portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and wherein the stem portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises the amino acid sequence of SEQ ID NO: 98, wherein the target-binding domain “T” of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and wherein the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98.

[0421] In some embodiments, the polypeptide containing formula T-S1 provided herein (where T is a polypeptide containing V) H and V L The target-binding domain (and S1 is the stem portion) contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain HThe V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L The amino acids 172-283 of SEQ ID NO: 98 correspond to amino acids 284-634 of SEQ ID NO: 98, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein contains at least 90% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L The amino acids 172-283 of SEQ ID NO: 98 correspond to amino acids 284-634 of SEQ ID NO: 98, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein contains at least 95% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L The amino acids 172-283 of SEQ ID NO: 98 correspond to amino acids 284-634 of SEQ ID NO: 98, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein contains at least 98% identical amino acid sequence to the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L The amino acids 172-283 of SEQ ID NO: 98 correspond to amino acids 284-634 of SEQ ID NO: 98, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising formula T-S1 provided herein comprises the amino acid sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L The amino acids 172-283 of SEQ ID NO: 98 correspond to amino acids 284-634 of SEQ ID NO: 98, and the stem portion “S1” of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98.

[0422] In some embodiments, the polypeptide of the formula T-L1-Fc-L2-X1 provided herein (where T is a target-binding domain, L1 is a polypeptide linker or is absent, Fc is a variant Fc domain as provided herein, L2 is a polypeptide linker or is absent, and X1 is a polypeptide containing a transmembrane domain) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the sequence of SEQ ID NO: 98, wherein the target-binding domain “T” of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 comprises amino acids 284-301 of SEQ ID NO: 98, Fc comprises amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising the formula T-L1-Fc-L2-X1 provided herein contains at least 90% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the target-binding domain “T” of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98. In some embodiments, the polypeptide of formula T-L1-Fc-L2-X1 provided herein contains at least 95% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98. In some embodiments, the polypeptide of formula T-L1-Fc-L2-X1 provided herein contains at least 98% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98.In some embodiments, the polypeptide of formula T-L1-Fc-L2-X1 provided herein contains the amino acid sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98.

[0423] In some embodiments, the polypeptide comprising the formula T-L1-Fc-L2-X1 provided herein (where T is a polypeptide comprising V) H and V L The target-binding domain, L1 is a peptide linker or is absent, Fc is a variant Fc domain as provided herein, L2 is a peptide linker or is absent, and X1 is a peptide containing a transmembrane domain) comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequence to the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising the formula T-L1-Fc-L2-X1 provided herein contains at least 90% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising the formula T-L1-Fc-L2-X1 provided herein contains at least 95% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain... HThe V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising the formula T-L1-Fc-L2-X1 provided herein contains at least 98% identical amino acid sequence to the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98. In some embodiments, the polypeptide comprising the formula T-L1-Fc-L2-X1 provided herein contains the amino acid sequence of SEQ ID NO: 98, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L The amino acids 172-283 of SEQ ID NO: 98 correspond to L1, which contains amino acids 284-301 of SEQ ID NO: 98, and Fc which contains amino acids 302-542 of SEQ ID NO: 98. L2 is absent, and X1 corresponds to amino acids 543-634 of SEQ ID NO: 98.

[0424] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD peptides (where T is the target-binding domain, L1 is the peptide linker or absent, Fc is the variant Fc domain as provided in this paper, L2 is the peptide linker or absent, ECD is the extracellular domain, T M(This is a transmembrane domain, and the ICD is an intracellular domain containing an env-incorporated motif) contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains at least 90% identical amino acid sequence to that of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains at least 95% identical amino acid sequence to that of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... MThe ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains at least 98% identical amino acid sequence to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of -ICD contains the amino acid sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98.

[0425] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD polypeptide (where T is the peptide containing V) H and V L The target-binding domain, L1 is a peptide linker or is absent, Fc is the variant Fc domain as provided in this paper, L2 is a peptide linker or is absent, ECD is an extracellular domain, T MIt is a transmembrane domain, and the ICD is an intracellular domain containing an env-incorporated motif. It contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 90% of the same amino acid sequence as SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of -ICD contains at least 95% identical amino acid sequence to the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. LCorresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 98% of the same amino acid sequence as SEQ ID NO:98, wherein the V target-binding domain... H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains the amino acids of SEQ ID NO: 98, wherein the V of the target-binding domain H The V-type target-binding domain corresponds to amino acids 25-150 of SEQ ID NO: 98. L Corresponding to amino acids 172-283 of SEQ ID NO: 98, L1 contains amino acids 284-301 of SEQ ID NO: 98, Fc contains amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains amino acids 585-612 of SEQ ID NO: 98, and amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains amino acids 627-634 of SEQ ID NO: 98.

[0426] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD peptides (where T is the target-binding domain, L1 is the peptide linker or absent, Fc is the variant Fc domain as provided in this paper, L2 is the peptide linker or absent, ECD is the extracellular domain, T M The transmembrane domain, and the ICD (an intracellular domain containing an env-incorporated motif), contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide contains the amino acid sequence SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98, Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98, and T... M The ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide contains the amino acid sequence SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98, Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98, T MThe ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide contains the amino acid sequence SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98, Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98, T M The ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of the -ICD contains an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide contains the amino acid sequence SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98, Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98, T MThe ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and the ICD contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The polypeptide of -ICD contains the amino acid sequence of SEQ ID NO: 98, wherein the target-binding domain "T" of the polypeptide contains the amino acid sequence SEQ ID NO: 39 and corresponds to amino acids 25-283 of SEQ ID NO: 98, L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98, Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98, T M The amino acid sequence comprising SEQ ID NO: 62 and corresponding to amino acids 585-612 of SEQ ID NO: 98, and the ICD comprising amino acids 613-634 of SEQ ID NO: 98, wherein the env incorporation motif comprises the amino acid sequence comprising SEQ ID NO: 63 and corresponding to amino acids 627-634 of SEQ ID NO: 98.

[0427] In some implementations, the included T-L1-Fc-L2-ECD-T provided herein M -ICD polypeptide (where T is the peptide containing V) H and V L The target-binding domain, L1 is a peptide linker or is absent, Fc is the variant Fc domain as provided in this paper, L2 is a peptide linker or is absent, ECD is an extracellular domain, T M It is a transmembrane domain, and the ICD is an intracellular domain containing an env-incorporated motif. It contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences to the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain... H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 98, V of the target-binding domain.L The sequence contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 98; L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98; Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98; L2 is absent; ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98; T M The ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 90% of the same amino acid sequence as SEQ ID NO: 98, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 98, V of the target-binding domain. L The sequence contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 98; L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98; Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98; L2 is absent; ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98; T M The ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... MThe -ICD polypeptide contains at least 95% of the same amino acid sequence as SEQ ID NO: 98, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 98, V of the target-binding domain. L The sequence contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 98; L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98; Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98; L2 is absent; ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98; T M The ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains at least 98% of the same amino acid sequence as the sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 98, V of the target-binding domain. L The sequence contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 98; L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98; Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98; L2 is absent; ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98; T MThe ICD contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98, and contains amino acids 613-634 of SEQ ID NO: 98, wherein the env-incorporated motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the inclusion formula T-L1-Fc-L2-ECD-T provided herein is... M The -ICD polypeptide contains the amino acid sequence of SEQ ID NO: 98, wherein the V of the target-binding domain H Contains the amino acid sequence of SEQ ID NO: 36 and corresponds to amino acids 25-150 of SEQ ID NO: 98, V of the target-binding domain. L The sequence contains the amino acid sequence of SEQ ID NO: 37 and corresponds to amino acids 172-283 of SEQ ID NO: 98; L1 contains the amino acid sequence of SEQ ID NO: 72 and corresponds to amino acids 284-301 of SEQ ID NO: 98; Fc contains the amino acid sequence of SEQ ID NO: 104 and corresponds to amino acids 302-542 of SEQ ID NO: 98; L2 is absent; ECD contains the amino acid sequence of SEQ ID NO: 60 and corresponds to amino acids 543-584 of SEQ ID NO: 98; T M The amino acid sequence comprising SEQ ID NO: 62 and corresponding to amino acids 585-612 of SEQ ID NO: 98, and the ICD comprising amino acids 613-634 of SEQ ID NO: 98, wherein the env incorporation motif comprises the amino acid sequence comprising SEQ ID NO: 63 and corresponding to amino acids 627-634 of SEQ ID NO: 98.

[0428] In some implementations, V having a sequence as shown in SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD and T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H peptides and V having the sequence shown in SEQ ID NO:49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes CD8 and / or CD28 ECD, T MAnd ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, CD8, and / or CD28 T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. M And ICD. In some embodiments, V having a sequence as shown in SEQ ID NO: 48 H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD and T. M and ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 48. H peptides and V having the sequence shown in SEQ ID NO:49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes CD8 and / or CD28 ECD, T M and ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 48. H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) that includes ECD, CD8, and / or CD28 T. M and ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, V has a sequence as shown in SEQ ID NO: 48. H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide is linked to a stem portion (S1) containing an Fc region (L1-Fc-L2-X1) comprising CD8 and / or CD28 ECD, CD8 and / or CD28 T. Mand ICD, wherein the ICD contains an env-incorporated motif as provided herein. In some embodiments, a V motif having the sequence shown in SEQ ID NO: 48 is attached to the stem portion (S1) containing the Fc region (L1-Fc-L2-X1). H Peptide and V having the sequence shown in SEQ ID NO: 49 L The peptide anchors to the surface of viral particles (such as those described in this article), and this Fc region contains ECD, T... M and ICD. In some implementations, L1, Fc, L2, ECD, T M The identity of the ICD is as provided herein. In some implementations, V H and V L Peptides bind to immune cells (such as those described in this article).

[0429] In some embodiments, V comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO:49. L The peptide is linked to a stem portion (S1) containing the Fc region as described herein. In some embodiments, a V-shaped portion comprises at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L The peptide is linked to a stem portion (S1) comprising an Fc region that also contains a transmembrane domain. In some embodiments, the Fc region, which also contains a transmembrane domain, has the formula L1-Fc-L2-X1, where L1 is a linker as provided herein or is absent, Fc is a variant Fc region as provided herein, L2 is a linker as provided herein or is absent, and X1 is a polypeptide containing a transmembrane domain as provided herein. As provided herein, X1 may comprise a peptide having the formula ECD-T M-ICD polypeptide, wherein ECD is an extracellular domain or a fragment thereof as provided herein or is absent, TM is a transmembrane domain as provided herein, and ICD is an intracellular domain as provided herein or is absent. In some embodiments, V comprises a sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 49. L Peptide linked to a compound containing ECD, T M And the stem portion (S1) of the Fc region (L1-Fc-L2-X1) of the ICD. In some embodiments, V contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sequence identical to that of SEQ ID NO: 48. H Peptide; and V containing at least 85%, 86%, 87...

Claims

1. A particle or composition comprising one or more nucleic acid molecules encoding one of the following: a) Chimeric antigen receptor (CAR); and b) A polypeptide, wherein the polypeptide comprises: i) Ligand-binding extracellular domain (ECD); ii) Immune cell activation intracellular domain (ICD); and iii) A transmembrane structural domain connecting the ECD and the ICD.

2. The particle according to claim 1, wherein the immune cell activating ICD comprises an ICD of hGCSF-R or a mutant or fragment thereof.

3. The particle according to claim 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111, or an amino acid sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

4. The particle according to claim 3, wherein the hGCSF-R ICD does not contain the sequence of SEQ ID NO:

110.

5. The particle according to claim 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111 and does not contain amino acid residues 800-813, 775-813, 750-813, 725-813 or 716-813 of SEQ ID NO: 125 or fragments thereof located at the C-terminal residues of SEQ ID NO:

111.

6. The particle according to claim 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111 and amino acid residues 716-812 of SEQ ID NO: 125 or a fragment thereof located at the C-terminal residue of SEQ ID NO: 111, provided that the ICD does not contain a continuous fragment of amino acid residues 716-813 of SEQ ID NO:

125.

7. The particle according to claim 1, wherein the ECD is a GCSF-RECD, and optionally wherein the GCSF-RECD is a human GCSF-RECD containing the amino acid sequence of SEQ ID NO: 107 or SEQ ID NO:

108.

8. The particle of claim 1, wherein the ECD comprises an ECD of human growth hormone receptor (hGH-R) or a mutant or fragment thereof, wherein the ECD of human growth hormone receptor (hGH-R) comprises an amino acid sequence encoded by exons 1-7 of hGH-R or a mutant or fragment thereof.

9. The particle of claim 8, wherein the ECD of the hGH-R contains a mutation of C259R.

10. The particle according to claim 8, wherein the ECD of the hGH-R comprises the amino acid sequence of SEQ ID NO: 114 or SEQ ID NO: 115, or an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with it.

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