Activation of T cells by mutant G-CSFR

Abstract

This specification provides particles or compositions comprising one or more nucleic acid molecules encoding a polypeptide comprising a chimeric antigen receptor (CAR) and / or 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 same; and nucleic acid molecules encoding the same. This specification also provides compositions comprising the same and methods for using the same. This specification also provides viral particles comprising the same.

Description

[Technical Field]

[0001] (Related applications) This application claims the interests of U.S. Provisional Applications 63 / 508,055 (filed June 14, 2023) and U.S. Provisional Applications 63 / 510,745 (filed June 28, 2023), which are incorporated herein by reference in their entirety.

[0002] (Reference to electronically submitted sequence listings) This application includes a sequence listing submitted electronically in XML format, the entirety of which is incorporated herein by reference. The XML copy, created on June 5, 2024, is named "INH-023WO_SL.xml" and is 146,320 bytes in size.

[0003] (Field of invention) The embodiments provided herein relate to variant G-CSFR constructs and methods for using them. The embodiments provided herein also relate to viral particles comprising heterologous viral glycoproteins, a targeting moiety, and a nucleic acid molecule encoding the heterologous molecule of interest. In some embodiments, the nucleic acid molecule encodes a chimeric antigen receptor and a variant G-CSFR. The embodiments provided herein further relate to compositions comprising the viral particles provided herein and methods for using them. [Background technology]

[0004] Chimeric antigen receptor T (CAR-T) cell therapy has revolutionized antitumor treatment, particularly for hematological malignancies. However, significant limitations such as high cost, long manufacturing time, and safety concerns persist, hindering the broad 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 T cell targeting vectors. Conventional CAR-T methods require the isolation and proliferation of individualized T cells after CAR transduction before they can be injected back into the patient. Thus, one potential pitfall of in vivo CAR-T is the hypothetical lack of proliferation of CAR-containing T cells, which could limit the effectiveness of the method. This embodiment addresses these needs, as well as others. [Overview of the project]

[0005] In some embodiments, particles or compositions are provided. In some embodiments, the particles or compositions comprise one or more nucleic acid molecules encoding a polypeptide comprising 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.

[0006] In some embodiments, the immune cell activation ICD comprises an ICD of hGCSF-R (human granulocyte colony-stimulating factor receptor), or a variant 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 thereto.

[0007] 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.

[0008] In some embodiments, the ECD is a human growth hormone receptor (hGH-R) ECD, or a variant or fragment thereof. In some embodiments, the hGH-R ECD includes 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 thereto.

[0009] In some embodiments, the polypeptide comprises an ECD which is a GCSFR-ECD, or a variant thereof (e.g., as described herein); a transmembrane domain which is a GCSF-R transmembrane domain, or a variant thereof (e.g., as described herein); and an immunoactivated ICD which is a GCSF-R ICD, or a variant thereof (e.g., as described herein).

[0010] In some embodiments, the polypeptide comprises an ECD which is hGH-R-ECD, or a variant thereof (e.g., as described herein); a transmembrane domain which is a GCSF-R transmembrane domain, or a variant thereof (e.g., as described herein); and an immunoactivated ICD which is a GCSF-R ICD, or a variant thereof (e.g., as described herein).

[0011] In some embodiments, the polypeptide comprises an ECD which is hGH-R-ECD, or a variant thereof (e.g., as described herein); a transmembrane domain which is a gp130 transmembrane domain, or a variant thereof (e.g., as described herein); and an immunoactivated ICD containing a gp130 ICD, or a variant thereof (e.g., as described herein) and an IL-2Rβ ICD, or a variant thereof (e.g., as described herein).

[0012] In some embodiments, the polypeptide comprises an ECD which is hGH-R-ECD, or a variant thereof (e.g., as described herein); a transmembrane domain which is a GCSF-R transmembrane domain, or a variant thereof (e.g., as described herein); and a GCSF-R ICD, or a variant thereof (e.g., as described herein) and an IL-7RαICD, or a variant thereof (e.g., as described herein).

[0013] 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 any of the aforementioned variants. 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 respect to 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.

[0014] In some embodiments, a nucleic acid molecule is provided, which comprises i) a ligand-binding extracellular domain (ECD) provided herein; ii) an immune cell-activating intracellular domain (ICD) provided herein; and iii) a transmembrane domain that ligates the ECD to the ICD provided herein.

[0015] 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 the CD20 CAR provided herein. In some embodiments, the CAR comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 99, at least 90% identity to SEQ ID NO: 99, at least 95% identity to SEQ ID NO: 99, at least 99% identity to SEQ ID NO: 99, or at least 100% identity to 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 a 2A sequence.

[0016] In some embodiments, polypeptides are provided. In some embodiments, the polypeptide comprises i) a ligand-binding extracellular domain (ECD) provided herein; ii) an immune cell-activating intracellular domain (ICD) provided herein; and iii) a transmembrane domain ligating the ECD to the ICD provided herein.

[0017] In some embodiments, particles comprising polypeptides provided herein are provided. 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 vectors are pseudotyped viral vectors.

[0018] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety, and one or more nucleic acid molecules encoding one or more heterologous molecules of interest, wherein the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety, and the nucleic acid molecule encoding the heterologous molecule of interest is a nucleic acid molecule provided herein.

[0019] In some embodiments, the stalk portion S1 comprises a modified Fc protein. In some embodiments, the modified Fc protein comprises a transmembrane domain, such as but not limited to a CD8 or CD28 transmembrane domain. In some embodiments, the modified Fc protein comprises an effector mutation, which inhibits the interaction between the Fc protein and Fc-interacting proteins such as FcγR, C1q, FcRβ, or FcRn.

[0020] In some embodiments, the modified Fc protein is a modified IgG1 Fc protein containing one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A.

[0021] In some embodiments, the modified IgG1 Fc protein contains an amino acid sequence having at least 80% identity to SEQ ID NO: 104, at least 85% identity to SEQ ID NO: 104, at least 90% identity to SEQ ID NO: 104, at least 95% identity to SEQ ID NO: 104, at least 98% identity to SEQ ID NO: 104, or at least 100% identity to SEQ ID NO: 104.

[0022] In some embodiments, the modified Fc protein is a modified IgG2 Fc protein containing one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A.

[0023] In some embodiments, the modified Fc protein is a modified IgG4 Fc protein containing one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A.

[0024] In some embodiments, the targeting moiety having formula T-S1 comprises a stalk moiety S1 having formula L1-Fc-L2-X1, where L1 is a linker or absent, Fc is a modified Fc protein, L2 is a linker or absent, and X1 is a polypeptide containing a transmembrane domain, and the targeting moiety having formula T-S1 has formula T-L1-Fc-L2-X1.

[0025] In some embodiments, the polypeptide containing the transmembrane domain (X1) is ECD-T M - Contains a polypeptide having the formula ICD, where ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M ICD is the transmembrane domain of a transmembrane protein, and ICD is a protein that promotes the incorporation of the targeting portion into the intracellular domain or the envelope of a viral particle, or is absent; the targeting portion having the formula T-L1-Fc-L2-X1 is the formula T-L1-Fc-L2-ECD-T M - Has an ICD.

[0026] In some embodiments, the stalk portion S1 comprises formula L3-X1, where L3 is a flexible peptide linker and X1 is a polypeptide comprising a transmembrane domain; the targeting portion having formula T-S1 comprises formula T-L3-X1.

[0027] In some embodiments, the polypeptide containing the transmembrane domain (X1) is ECD-T M - Contains a polypeptide having the formula ICD, where ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent; T M ICD is the transmembrane domain of a transmembrane protein, and ICD is a protein that promotes the incorporation of the targeting portion into the intracellular domain or the envelope of a viral particle, or is absent; the targeting portion having formula T-L3-X1 is formula T-L3-ECD-T M - Has an ICD.

[0028] In some embodiments, the targeted portion binds to CD7. In some embodiments, the targeted portion includes a polypeptide having a heavy chain variable region including 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 targeted portion includes a polypeptide having a light chain variable region including 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 includes a heavy chain variable region having at least 90% sequence identity with SEQ ID NO: 36. In some embodiments, the light chain includes a light chain variable region having at least 90% sequence identity with SEQ ID NO: 37. In some embodiments, the targeted portion that binds to CD7 includes a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 38. In some embodiments, the targeted portion that binds to CD7 includes a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 39.

[0029] In some embodiments, the targeted portion binds to CD8. In some embodiments, the targeted portion includes a polypeptide having a heavy chain variable region containing 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 targeted portion includes a polypeptide having a light chain variable region containing LCDR1 of SEQ ID NO: 45, LCDR2 of SEQ ID NO: 46, and LCDR3 of SEQ ID NO: 47, or a variant thereof as described above. In some embodiments, the heavy chain includes a heavy chain variable region having at least 90% sequence identity with SEQ ID NO: 48. In some embodiments, the light chain includes a light chain variable region having at least 90% sequence identity with SEQ ID NO: 49. In some embodiments, the targeted portion that binds to CD8 includes a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 50. In some embodiments, the targeted portion that binds to CD8 includes a polypeptide having a sequence having at least 90% sequence identity with SEQ ID NO: 51.

[0030] In some embodiments, the heterologous viral glycoprotein is the SVCV-G polypeptide provided herein.

[0031] In some embodiments, the heterologous viral glycoprotein is a VSV-G polypeptide. In some embodiments, the VSV-G polypeptide includes 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.

[0032] In some embodiments, the viral particles provided herein further comprise a nucleic acid molecule encoding the 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.

[0033] In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence that is at least 95% identical to SEQ ID NO: 92. In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence that is at least 95% identical to SEQ ID NO: 93. In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence that is at least 95% identical to SEQ ID NO: 96. In some embodiments, the CAR includes an antigen-binding domain having an amino acid sequence that is at least 95% identical to SEQ ID NO: 97.

[0034] In some embodiments, the CAR includes 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.

[0035] In some embodiments, methods for infecting cells are provided. In some embodiments, the method includes bringing cells into contact with viral particles provided herein.

[0036] In some embodiments, methods are provided for infecting cells in a target. In some embodiments, the method includes administering a pharmaceutical composition comprising viral particles provided herein to the target.

[0037] In some embodiments, a method for delivering a target heterologous molecule to a cell is provided. In some embodiments, the method comprises contacting a cell with a viral particle provided herein, the viral particle comprising a nucleic acid molecule encoding the target heterologous molecule.

[0038] In some embodiments, methods are provided for delivering a target heterologous molecule to target cells. In some embodiments, the method comprises administering viral particles provided herein to the target, the viral particles comprising a nucleic acid molecule encoding the target heterologous molecule.

[0039] In some embodiments, methods are provided for treating a disease or disorder in a subject. In some embodiments, the method comprises administering viral particles provided herein to a subject, the viral particles comprising nucleic acid molecules encoding a heterogeneous molecule of interest for treating a disease or disorder.

[0040] In some embodiments, methods are provided for delivering heterologous molecules to target cells. In some embodiments, the method comprises contacting cells with viral particles provided herein, the viral particles comprising nucleic acid molecules encoding heterologous molecules.

[0041] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, the heterologous viral glycoprotein comprises a sequence selected from SEQ ID NOs: 22, 23, 24, 25, 52, or 53. In some embodiments, the target-binding domain comprises a sequence selected from SEQ ID NOs: 38, 39, 50, or 51. In some embodiments, the stalk moiety S1 comprises a modified Fc protein comprising a sequence that is a modified version of SEQ ID NOs: 26, 27, or 28. In some embodiments, the modified version of SEQ ID NOs: 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 contains 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 contains 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 includes 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 the nucleic acid molecule provided herein.

[0042] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is the target-binding domain and S1 is the stalk moiety. In some embodiments, the heterologous viral glycoprotein comprises a sequence selected from SEQ ID NOs: 22, 23, 24, 25, 52, or 53. In some embodiments, the target-binding domain comprises a sequence selected from SEQ ID NOs: 38, 39, 50, or 51. In some embodiments, the stalk portion S1 comprises the formula L1-Fc-L2-X1, where L1 is a linker containing or absent 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; Fc is a modified Fc protein containing a sequence that is a modified version of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28; L2 is a linker containing or absent 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; and X1 is a polypeptide containing a transmembrane domain. In some embodiments, the modified version 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 contains 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 contains one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A. In some embodiments, the polypeptide containing the transmembrane domain (X1) is ECD-T M - The formula is ICD, and 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; TM is a transmembrane domain having the sequence or fragment thereof of SEQ ID NO: 61 or SEQ ID NO: 62; ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, and ICD may include an env incorporation motif comprising the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or ICD may not be present. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is the nucleic acid molecule provided herein.

[0043] In some embodiments, virus particles are provided. In some embodiments, the virus particles comprise a heterologous viral glycoprotein, a targeting moiety 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 stalk portion. In some embodiments, the heterologous viral 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 stalk portion S1 comprises the formula L1-Fc-L2-X1, wherein L1 is a linker comprising the sequence of SEQ ID NO: 55 or is absent; Fc is a modified Fc protein comprising a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28, L2 is a linker comprising the sequence of SEQ ID NO: 55 or is absent; 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 ECD is an extracellular domain having the sequence of SEQ ID NO: 60, or a fragment thereof, or is absent; T Mis a transmembrane domain or fragment thereof having the sequence of SEQ ID NO: 62; ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, and ICD may include an env incorporation motif containing the amino acid sequence of SEQ ID NO: 63 or SEQ ID NO: 64, or ICD may not be present. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest is the nucleic acid molecule provided herein.

[0044] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is the target-binding domain and S1 is the stalk moiety. 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 SEQ ID NO: 39a. In some embodiments, the stalk moiety S1 comprises the formula L1-Fc-L2-X1, where L1 is a linker comprising the sequence of SEQ ID NO: 55; Fc is a modified Fc protein comprising the sequence of SEQ ID NO: 104; L2 is a linker and is absent; and X1 is a polypeptide comprising a transmembrane domain. In some embodiments, the polypeptide comprising the transmembrane domain (X1) is ECD-T M - The formula is ICD, and ECD is the extracellular domain having the sequence of sequence number 60; T M is a transmembrane domain having the sequence of SEQ ID NO: 62; ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and ICD includes an env incorporation motif having the amino acid sequence of SEQ ID NO: 63. In some embodiments, a nucleic acid molecule encoding a heterologous molecule of interest encodes a chimeric antigen receptor provided herein, and a polypeptide comprising i) a ligand-binding extracellular domain (ECD) provided herein, ii) an immune cell-activating intracellular domain (ICD) provided herein, and ii) a transmembrane domain linking the ECD to the ICD provided herein.

[0045] In some embodiments, a viral particle is provided, which comprises a heterologous viral glycoprotein, a targeting moiety, 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 to SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identity to SEQ ID NO: 23 or SEQ ID NO: 25, at least 99% identity to SEQ ID NO: 23 or SEQ ID NO: 25, or at least 100% identity to SEQ ID NO: 23 or SEQ ID NO: 25. In some embodiments, the targeting moiety comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 98, at least 95% identity to SEQ ID NO: 98, at least 99% identity to SEQ ID NO: 98, or at least 100% identity to SEQ ID NO: 98. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest encodes a chimeric antigen receptor provided herein, and a polypeptide comprising i) a ligand-binding extracellular domain (ECD) provided herein, ii) an immune cell-activating intracellular domain (ICD) provided herein, and ii) a transmembrane domain linking the ECD to the ICD provided herein.

[0046] In some embodiments, a viral particle is provided, which comprises a heterologous viral glycoprotein, a targeting moiety, 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 to SEQ ID NO: 52 or SEQ ID NO: 53, at least 95% identity to SEQ ID NO: 52 or SEQ ID NO: 53, at least 99% identity to SEQ ID NO: 52 or SEQ ID NO: 53, or at least 100% identity to SEQ ID NO: 52 or SEQ ID NO: 53. In some embodiments, the targeting moiety comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 98, at least 95% identity to SEQ ID NO: 98, at least 99% identity to SEQ ID NO: 98, or at least 100% identity to SEQ ID NO: 98. In some embodiments, the nucleic acid molecule encoding the heterologous molecule of interest encodes a chimeric antigen receptor provided herein, and a polypeptide comprising i) a ligand-binding extracellular domain (ECD) provided herein, ii) an immune cell-activating intracellular domain (ICD) provided herein, and ii) a transmembrane domain linking the ECD to the ICD provided herein.

[0047] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is a target-binding domain and S1 is a stalk moiety. 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 stalk moiety S1 comprises the formula L3-X1, where 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 polypeptide linker comprising a transmembrane domain. In some embodiments, the polypeptide comprising the transmembrane domain (X1) is ECD-T M-Having the formula ICD, ECD is either 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 comprising the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and the ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, and the ICD comprises an env incorporation motif comprising 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 provided herein.

[0048] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is a target-binding domain and S1 is a stalk moiety. 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 stalk moiety S1 comprises the formula L3-X1, where 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 polypeptide linker comprising a transmembrane domain. In some embodiments, the polypeptide comprising the transmembrane domain (X1) is ECD-T M -Having the formula ICD, ECD is either 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 comprising the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and the ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, and the ICD comprises an env incorporation motif comprising 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 provided herein.

[0049] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is a target-binding domain and S1 is a stalk moiety. 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 stalk moiety S1 comprises the formula L3-X1, where 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 polypeptide linker comprising a transmembrane domain. In some embodiments, the polypeptide comprising the transmembrane domain (X1) is ECD-T M -Having the formula ICD, ECD is either 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 comprising the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and the ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, and the ICD comprises an env incorporation motif comprising 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 provided herein.

[0050] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is a target-binding domain and S1 is a stalk moiety. 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 stalk moiety S1 comprises the formula L3-X1, where 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 comprising the transmembrane domain (X1) is ECD-T M -Having the formula ICD, ECD is either 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 comprising the amino acid sequence of SEQ ID NO: 61 or SEQ ID NO: 62, and the ICD is an intracellular domain or protein that facilitates the incorporation of the targeting portion into the envelope of the viral particle, and the ICD comprises an env incorporation motif comprising 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 provided herein.

[0051] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is a target-binding domain and S1 is a stalk moiety. 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 stalk moiety S1 comprises the formula L3-X1, where 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 comprising the transmembrane domain (X1) is ECD-T M - It has the formula ICD, and ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59, T M ICD is a transmembrane domain comprising the amino acid sequence of SEQ ID NO: 61, and ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and ICD comprises an env incorporation motif comprising 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 provided herein.

[0052] In some embodiments, viral particles are provided. In some embodiments, the viral particle comprises a heterologous viral glycoprotein, a targeting moiety comprising a polypeptide having the formula T-S1, and a nucleic acid molecule encoding a heterologous molecule of interest, where T is a target-binding domain and S1 is a stalk moiety. 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 stalk moiety S1 comprises the formula L3-X1, where 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 comprising the transmembrane domain (X1) is ECD-T M - It has the formula ICD, and ECD is an extracellular domain containing the amino acid sequence of SEQ ID NO: 59, T M The ICD is a transmembrane domain comprising the amino acid sequence of SEQ ID NO: 61, and the ICD is an intracellular domain or protein that facilitates the incorporation of a targeting portion into the envelope of a viral particle, and the ICD comprises an env incorporation motif comprising 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 provided herein, and a polypeptide comprising i) a ligand-binding extracellular domain (ECD) provided herein, ii) an immune cell-activating intracellular domain (ICD) provided herein, and ii) a transmembrane domain linking the ECD to the ICD provided herein.

[0053] In some embodiments, the CAR includes 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. [Brief explanation of the drawing]

[0054] [Figure 1A] This specification shows exemplary chimeric GCSFR structures. [Figure 1B] This specification shows exemplary chimeric GCSFR structures.

[0055] [Figure 2A] Figure 2A shows the crystal structure of VSV-G bound to LDL-R. [Figure 2B] Figure 2B shows the crystal structure of VSV-G bound to LDL-R.

[0056] [Figure 3A] The effect of adding negatively charged amino acids to VSV-G is demonstrated. LDL-R binding interacts with innate affinity and fusion properties. Figure 3A shows the titration of the VSV-G construct on SupT1 cells. [Figure 3B] The effect of adding negatively charged amino acids to VSV-G is demonstrated. The LDL-R bond interacts with the innate affinity and fusion properties. Figure 3B shows the functional titer of each construct calculated from the titration in Figure 3A.

[0057] [Figure 4-1] This shows the alignment of ectodomains of different VSV-G proteins from different strains. [Figure 4-2] This shows the alignment of ectodomains of different VSV-G proteins from different strains. [Figure 4-3] This shows the alignment of ectodomains of different VSV-G proteins from different strains. [Figure 4-4] This shows the alignment of ectodomains of different VSV-G proteins from different strains.

[0058] [Figure 5]This study demonstrates the effects of various VSV-G mutations on the serum stability of viral constructs combined with a CD7 binder.

[0059] [Figure 6] This study demonstrates the effects of various VSV-G mutations on the serum stability of viral constructs combined with a CD7 binder.

[0060] [Figure 7] The ability of various rhabdoviral G proteins to transduce SupT1 and PBMC cells, either alone or in combination with a CD7 binder, is demonstrated.

[0061] [Figure 8A] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8B] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8C] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8D] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8E] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8F] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8G] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8H]Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8I] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8J] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8K] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 8L] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0062] [Figure 8M] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0063] [Figure 9A] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9B] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9C] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9D] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9E]Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9F] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9G] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9H] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9I] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9J] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9K] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown. [Figure 9L] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0064] [Figure 9M] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD7 binder disclosed herein are shown.

[0065] [Figure 10A] Flow cytometry data of human PBMCs transduced with exemplary vectors containing the CD8 binder disclosed herein are shown.

[0066] [Figure 10B] Flow cytometry data of non-human primate PBMCs transduced with exemplary vectors containing the CD8 binder disclosed herein are shown.

[0067] [Figure 11A] This demonstrates the ability of VSV-G* pseudotyped lentivirus particles containing a modified Fc stalk and a CD7 binder to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 11B] This shows cell transduction in the absence of a CD7 binder. VSV-G* indicates VSV-G(I182E, T214N, T352A). [Figure 11C] The MOI calculated from SupT1 titrations shows the transduction of human and non-human primate PBMCs. VSV-G* indicates VSV-G(I182E, T214N, T352A).

[0068] [Figure 12A] This demonstrates the ability of VSV-G* pseudotyped lentivirus particles, possessing a CD7 binder with a flexible stalk, to transduce SupT1 cells and human and non-human primate PBMCs. [Figure 12B] This demonstrates cell transduction in the absence of a CD7 binder. [Figure 12C] The transduction of human and non-human primate PBMCs is shown from the perspective of MOI calculated from SupT1 titration. VSV-G* refers to VSV-G(I182E, T214N, T352A).

[0069] [Figure 13A] This study demonstrates the ability of viral particles containing CD7 binders with flexible stalks of varying lengths to transduce SupT1 cells, compared to other IgG-based binders. [Figure 13B]This study demonstrates the ability of viral particles containing CD7 binders with flexible stalks of varying lengths to transduce activated PBMC cells, compared to other IgG-based binders.

[0070] [Figure 14A] This demonstrates the ability of SVCV-G pseudotyped lentivirus particles, possessing a modified Fc-stalk CD7 binder, to transduce SupT1 cells and human and non-human primate PBMCs. [Figure 14B] This demonstrates cell transduction in the absence of a CD7 binder. [Figure 14C] The MOI calculated from SupT1 titrations shows the transduction of human and non-human primate PBMCs.

[0071] [Figure 15A] This demonstrates the ability of SVCV-G pseudotyped lentiviral particles, possessing a CD7 binder with a flexible stalk, to transduce SupT1 cells and human and non-human primate PBMCs. [Figure 15B] This demonstrates cell transduction in the absence of a CD7 binder. [Figure 15C] This study demonstrates the transduction of human and non-human primate PBMCs from the perspective of MOI calculated from SupT1 titration.

[0072] [Figure 16A] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 16B] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 16C]This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 16D] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A).

[0073] [Figure 17A] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 17B] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 17C] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 17D] This study compares the ability of VSV-G* pseudotyped lentiviral particles (Panels A and B) and SVCV-G pseudotyped lentiviral particles (Panels C and D) to transduce SupT1 cells and human and non-human primate PBMCs. VSV-G* refers to VSV-G(I182E, T214N, T352A).

[0074] [Figure 18A] This is a comparison of off-target transduction of GFP in a panel of B cell lines compared to control SupT1 cells. Figure 18A shows data for VSV-G* pseudotyped lentiviral particles with a CD7 binder containing a modified Fc stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 18B] This is a comparison of off-target transduction of GFP in a panel of B cell lines compared to control SupT1 cells. Figure 18B shows data for SVCV-G pseudotyped lentivirus particles with a CD7 binder containing a modified Fc stalk.

[0075] [Figure 19A] This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to control SupT1 cells. Figure 19A shows data for VSV-G* pseudotyped lentiviral particles with a CD7 binder containing a modified Fc stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 19B] This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to control SupT1 cells. Figure 19B shows data for SVCV-G pseudotyped lentiviral particles with a CD7 binder containing a modified Fc stalk.

[0076] [Figure 20A] This is a comparison of off-target transduction of GFP in a panel of B cell lines compared to the control SupT1. Figure 20A shows data for VSV-G* pseudotyped lentiviral particles with a CD7 binder having a flexible stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 20B] This is a comparison of off-target transduction of GFP in a panel of B cell lines compared to the control SupT1. Figure 20B shows data for SVCV-G pseudotyped lentiviral particles with a CD7 binder having a flexible stalk.

[0077] [Figure 21A] This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to control SupT1. Figure 21A shows data for VSV-G* pseudotyped lentiviral particles with a CD7 binder containing a flexible stalk. VSV-G* refers to VSV-G(I182E, T214N, T352A). [Figure 21B] This is a comparison of off-target transduction of CAR20-T2A-GFP constructs in a panel of B cell lines compared to control SupT1. Figure 21B shows data for SVCV-G pseudotyped lentiviral particles with a CD7 binder containing a flexible stalk.

[0078] [Figure 22A] This specification demonstrates the ability of VSV-G* pseudotyped lentiviral particles utilizing the CD7 binder and CD20-CAR transgene provided herein to kill Daudi lymphoma cells (Figure 22A) or Raji lymphoma cells (Figure 22B). The CAR20 constructs utilized the antigen-binding domain provided herein, which includes rituximab or CD20AB1 SEQ ID NO: 92 or 93. [Figure 22B] This specification demonstrates the ability of VSV-G* pseudotyped lentiviral particles utilizing the CD7 binder and CD20-CAR transgene provided herein to kill Daudi lymphoma cells (Figure 22A) or Raji lymphoma cells (Figure 22B). The CAR20 constructs utilized the antigen-binding domain provided herein, which includes rituximab or CD20AB1 SEQ ID NO: 92 or 93.

[0079] [Figure 23A] This specification demonstrates the toxicity of ligand GCSF to PBMCs transduced with the CAR20-2A-chimeric GCSFR construct provided herein. Figure 23A shows the results for the control without the chimeric receptor. [Figure 23B]This specification demonstrates the toxicity of ligand GCSF to PBMCs transduced with the CAR20-2A-chimeric GCSFR construct provided herein. Figure 23B shows the results for GCSFR-d15. [Figure 23C] This specification demonstrates the toxicity of ligand GCSF to PBMCs transduced with the CAR20-2A-chimeric GCSFR construct provided herein. Figure 23C shows the results for G7R1. [Figure 23D] This specification demonstrates the toxicity of ligand GCSF to PBMCs transduced with the CAR20-2A-chimeric GCSFR construct provided herein. Figure 23D shows the results for G21 / R1. [Figure 23E] This specification demonstrates the toxicity of ligand GCSF to PBMCs transduced with the CAR20-2A-chimeric GCSFR construct provided herein. Figure 23E shows the results for G2R2. [Figure 23F] This specification demonstrates the toxicity of ligand GCSF to PBMCs transduced with the CAR20-2A-chimeric GCSFR construct provided herein. Figure 23F shows the results for G27 / 2R1.

[0080] [Figure 24A] The toxicity of ligand hGH to transduced PBMCs with the CAR20-2A-chimerin GHR construct provided herein is shown. Figure 24A shows the results for GHR-GCSFRd715. [Figure 24B] The toxicity of ligand hGH to transduced PBMCs with the CAR20-2A-chimerin GHR construct provided herein is shown. Figure 24B shows the results for GHR-G7R1. [Figure 24C] The toxicity of ligand hGH to transduced PBMCs with the CAR20-2A-chimerin GHR construct provided herein is shown. Figure 24C shows the results for GHR-G21 / 7R1. [Figure 24D]The toxicity of ligand hGH to transduced PBMCs with the CAR20-2A-chimerin GHR construct provided herein is shown. Figure 24D shows the results for GHR-G2R2. [Figure 24E] The toxicity of ligand hGH to transduced PBMCs with the CAR20-2A-chimerin GHR construct provided herein is shown. Figure 24E shows the results for GHR-G27 / 2R1.

[0081] [Figure 25A] This specification demonstrates the ability of the chimeric GCSFR receptor provided herein to induce proliferation of transduced PBMCs. Figure 25A shows the results for the control without the chimeric receptor. [Figure 25B] This specification demonstrates the ability of the chimeric GCSFR receptor provided herein to induce proliferation of transduced PBMCs. Figure 25B shows the results for GCSFR-d15. [Figure 25C] This specification demonstrates the ability of the chimeric GCSFR receptor provided herein to induce proliferation of transduced PBMCs. Figure 25C shows the results for G7R1. [Figure 25D] This specification demonstrates the ability of the chimeric GCSFR receptor to induce proliferation of transduced PBMCs. Figure 25D shows the results for G21 / R1. [Figure 25E] This specification demonstrates the ability of the chimeric GCSFR receptor provided herein to induce proliferation of transduced PBMCs. Figure 25E shows the results for G2R2. [Figure 25F] This specification demonstrates the ability of the chimeric GCSFR receptor provided herein to induce proliferation of transduced PBMCs. Figure 25F shows the results for G27 / 2R1.

[0082] [Figure 26A] This specification demonstrates the ability of the chimeric GHR receptor provided herein to induce proliferation of transduced PBMCs. Figure 26A shows the results for GHR-GCSFRd715. [Figure 26B]This specification demonstrates the ability of the chimeric GHR receptor provided herein to induce proliferation of transduced PBMCs. Figure 26B shows the results for GHR-G7R1. [Figure 26C] This specification demonstrates the ability of the chimeric GHR receptor provided herein to induce proliferation of transduced PBMCs. Figure 26C shows the results for GHR-G21 / 7R1. [Figure 26D] This specification demonstrates the ability of the chimeric GHR receptor provided herein to induce proliferation of transduced PBMCs. Figure 26D shows the results for GHR-G2R2. [Figure 26E] This specification demonstrates the ability of the chimeric GHR receptor provided herein to induce proliferation of transduced PBMCs. Figure 26E shows the results for GHR-G27 / 2R1.

[0083] [Figure 27A] The results of tracking experiments on PBMC-induced proliferation using selected chimeric receptor constructs are shown. Figure 27A shows the results for GCSFR FL. [Figure 27B] The results of tracking experiments on PBMC-induced proliferation using selected chimeric receptor constructs are shown. Figure 27B shows the results for GCSFRd715. [Figure 27C] The results of tracking experiments on PBMC-induced proliferation using selected chimeric receptor constructs are shown. Figure 27C shows the results for GCSFR-G2R2. [Figure 27D] The results of tracking experiments on PBMC-induced proliferation using selected chimeric receptor constructs are shown. Figure 27D shows the results for GHR-GCSFR. [Figure 27E] The results of tracking experiments on PBMC-induced proliferation using selected chimeric receptor constructs are shown. Figure 27E shows the results for GHR-dFN-GCSFR. [Figure 27F] The results for GHR-G2R2 are shown below.

[0084] [Figure 28A] The VSV-G* pseudotyped lentiviral particles utilizing the CD7 binder and CD20-CAR transgene provided herein demonstrate the ability to deplete B cells in vivo in huCD34 NSG mice. B cells were detected via evaluation for CD20 (Figure 28A) or CD19 (Figure 28B). [Figure 28B] The VSV-G* pseudotyped lentiviral particles utilizing the CD7 binder and CD20-CAR transgene provided herein demonstrate the ability to deplete B cells in vivo in huCD34 NSG mice. B cells were detected via evaluation for CD20 (Figure 28A) or CD19 (Figure 28B).

[0085] [Figure 29A] This specification demonstrates the ability of VSV-G* pseudotyped lentiviral particles, utilizing the CD7 binder and CD20-CAR transgene provided herein, to prevent tumorigenesis in vivo. Figure 29A shows the experimental design. [Figure 29B] This specification demonstrates the ability of VSV-G* pseudotyped lentiviral particles, utilizing the CD7 binder and CD20-CAR transgene provided herein, to prevent tumor formation in vivo. Figure 29B shows that mice intravenously received the lentiviral particles provided herein before Raji tumor injection had significantly lower tumor loads than mice receiving a control (GFP) vector or untreated mice. Tumor loads were measured by IVIS imaging.

[0086] [Figure 30A] This specification demonstrates the ability of VSV-G* pseudotyped lentiviral particles, utilizing the CD7 binder and CD20-CAR transgene provided herein, to eliminate established Raji tumors in vivo. Figure 30A shows the experimental design. [Figure 30B]This specification demonstrates the ability of VSV-G* pseudotyped lentiviral particles, utilizing the CD7 binder and CD20-CAR transgene provided herein, to eliminate established Raji tumors in vivo. Figure 30B shows that mice that received the lentiviral particles provided herein intravenously 6 days after Raji tumor injection had tumor volume reduced to below the detection limit. Tumor volume measured by IVIS imaging.

[0087] [Figure 31A] This shows an exemplary CAR-GCSF receptor construct for regulated cell proliferation. [Figure 31B] Flow cytometry data demonstrates controllable CAR cell proliferation in vitro. [Figure 31C] This shows the dose-response of CAR-T cells to exogenous GCSF agonists. [Figure 31D] This document presents an exemplary in vivo experimental design for evaluating CAR-GSCFR constructs. [Figure 31E] This document presents an exemplary in vivo experimental design for evaluating CAR-GSCFR constructs. [Figure 31F] This shows significant proliferation of CAR20+ cells in the presence of a GCSF agonist. [Figure 31G] This shows significant proliferation of CAR20+ cells in the presence of a GCSF agonist.

[0088] [Figure 32A] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GCSFR construct, or GFP after treatment with the indicated concentrations of GCSF or complete medium are shown. Figure 32A shows the proliferation of PBMCs from PBMC donor 1 transduced with CAR20 alone. [Figure 32B] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GCSFR construct, or GFP after treatment with the indicated concentrations of GCSF or complete medium are shown. Figure 32B shows the proliferation of PBMCs from PBMC donor 2 transduced with CAR20 alone. [Figure 32C] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GCSFR construct, or GFP after treatment with the indicated concentrations of GCSF or complete medium are shown. Figure 32C shows the proliferation of PBMCs from PBMC donor 1 transduced with CAR20 and GCSFR constructs. [Figure 32D] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GCSFR construct, or GFP after treatment with the indicated concentrations of GCSF or complete medium are shown. Figure 32D shows the proliferation of PBMCs from PBMC donor 2 transduced with CAR20 and GCSFR constructs. [Figure 32E] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GCSFR construct, or GFP after treatment with the indicated concentrations of GCSF or complete medium are shown. Figure 32E shows the proliferation of PBMCs from PBMC donor 1 transduced with GFP and GCSFR constructs. [Figure 32F] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GCSFR construct, or GFP after treatment with the indicated concentrations of GCSF or complete medium are shown. Figure 32F shows the proliferation of PBMCs from PBMC donor 2 transduced with GFP and GCSFR constructs.

[0089] [Figure 33A] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GHR construct, or GFP after treatment with GH or complete medium at the indicated concentrations are shown. Figure 33A shows the proliferation of PBMCs from PBMC donor 1 transduced with CAR20 alone. [Figure 33B] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GHR construct, or GFP after treatment with GH or complete medium at the indicated concentrations are shown. Figure 33B shows the proliferation of PBMCs from PBMC donor 2 transduced with CAR20 alone. [Figure 33C]The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GHR construct, or GFP after treatment with GH or complete medium at the indicated concentrations are shown. Figure 33C shows the proliferation of PBMCs from PBMC donor 1 transduced with CAR20 and GHR constructs. [Figure 33D] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GHR construct, or GFP after treatment with GH or complete medium at the indicated concentrations are shown. Figure 33D shows the proliferation of PBMCs from PBMC donor 2 transduced with CAR20 and GHR constructs. [Figure 33E] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GHR construct, or GFP after treatment with GH or complete medium at the indicated concentrations are shown. Figure 33E shows the proliferation of PBMCs from PBMC donor 1 transduced with GFP and GHR constructs. [Figure 33F] The results of cell count analysis of PBMCs transduced with a combination of CAR20 construct, GHR construct, or GFP after treatment with GH or complete medium at the indicated concentrations are shown. Figure 33F shows the proliferation of PBMCs from PBMC donor 2 transduced with GFP and GHR constructs. [Modes for carrying out the invention]

[0090] This specification provides particles or compositions comprising a chimeric antigen receptor (CAR) and one or more nucleic acid molecules encoding 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. As provided herein, the particles may be viral particles. Thus, viral particles comprising a heterologous viral glycoprotein, a targeting moiety, and one or more nucleic acid molecules encoding one or more heterologous molecules of interest are also provided herein, where the targeting moiety comprises a polypeptide having the formula T-S1, where T is the target-binding domain and S1 is the stalk moiety, and the 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-activating 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 the transmembrane domain provided herein. In some embodiments, S1 comprises a flexible polypeptide 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.

[0091] Furthermore, the viral particles may contain a VSV-G protein, which can be used to pseudotype viruses, such as lentiviruses. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of the New Jersey virus strain for bullous stomatitis, the Indiana virus strain for bullous stomatitis, the Alagoas virus strain for bullous stomatitis, the Maraba virus strain for bullous stomatitis, or the Carajas virus strain for bullous stomatitis. Examples of such proteins are provided herein.

[0092] Pseudotyped viruses, including those provided herein, which contain mutant VSV-G proteins, can be used in conjunction with a targeting moiety to facilitate the fusion of the pseudotyped virus with specific cells or tissues based on the expression of targets on those cells or tissues. As provided herein, the targeting moiety can be linked to an Fc protein, which may be called a stalk protein, which contains a transmembrane domain to facilitate the binding of the targeting moiety to the surface of the virus. In some embodiments, the Fc protein includes an Fc effector mutation, such as those provided herein.

[0093] Unless otherwise defined, all technical and scientific terms have the same meanings as those commonly understood by those skilled in the art in the field to which the disclosed embodiments belong.

[0094] As used herein, the terms "a" or "an" mean "at least one" or "one or more" unless the context clearly indicates otherwise.

[0095] Where used herein, the term “about” means that a number is an approximation and small variations will not significantly affect the implementation of the disclosed embodiment. Where numerical limitations are used, unless the context indicates otherwise, “about” means that the number may vary by ±10% and remain within the scope of the disclosed embodiment. Furthermore, where a phrase describes “about x to y,” the term “about” can be used interchangeably with the phrase “about x to y” unless the context indicates otherwise, modifying both x and y.

[0096] As used herein, the terms “individual,” “subject,” or “patient,” used interchangeably, mean any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates such as humans.

[0097] As used herein, the terms “comprising” (and any other form of “comprising,” such as “comprise,” “comprises,” and “comprised”), “having” (and any other form of “having,” such as “have,” and “has”), “including” (and any other form of “includings,” and “include”), or “containing” (and any other form of “containing,” such as “contains,” and “contain”) are inclusive or non-exclusive and do not exclude additional unlisted elements or process steps. Any process or composition using the transitional phrase “comprise” or “comprising” can be said to describe the same thing as the transitional phrase “consisting of” or “consists.”

[0098] As used herein, the term “contact” means bringing two elements together in an in vitro or in vivo setting. For example, “contact” of a virus or vector described herein with an individual, patient, or cells includes administering the virus to an individual or patient, such as a human, and introducing a compound into, for example, a sample containing cells or a purified preparation containing cells.

[0099] As used herein, the terms “fused” or “linked,” when used in reference to proteins having different domains or heterogeneous sequences, mean that the protein domains are part of the same peptide chain, connected to each other by either peptide bonds or other covalent bonds. Domains or sections may be directly linked or fused to each other, or another domain or peptide sequence may exist between the two domains or sequences, and such sequences are considered to be fused or linked to each other. In some embodiments, the various domains or proteins provided herein are either directly linked or fused to each other, or a linker sequence, such as a glycine / serine sequence described herein, links the two domains together.

[0100] A "disease" is a state of animal health in which the animal is unable to maintain homeostasis, and if the disease does not improve, the animal's health continues to deteriorate. In contrast, a "disorder" in animals is a state of health in which the animal is able to maintain homeostasis, but the animal's health is less desirable than in the absence of the disorder. Leaving a disorder untreated does not necessarily lead to a further decline in the animal's health.

[0101] "Effective dose" or "therapeutic effective dose" is used interchangeably herein and refers to the amount of a compound, formulation, material, or composition described herein that is effective in achieving a particular biological outcome or providing a therapeutic or preventive benefit. Such an outcome may include, but is not limited to, an amount that, when administered to a mammal, elicits a detectable level of immune cell activation compared to the immune cell activation detected in the absence of the composition. The immune response can be readily assessed by many methods recognized in the art. Those skilled in the art will understand that the amount of composition administered herein may vary and can be readily determined based on several factors, such as the disease or condition being treated, the age and health and physical condition of the mammal being treated, the severity of the disease, and the particular compound being administered.

[0102] "Code" refers to the inherent properties of a particular sequence of nucleotides in a polynucleotide (e.g., a gene, cDNA, or mRNA) and the resulting biological properties, which serve as a template for the synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA, and mRNA) or a defined sequence of amino acids. Thus, a gene codes for a protein if the transcription and translation of the mRNA corresponding to that gene produces a protein in a cell or other biological system. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is usually provided in a sequence listing, and the non-coding strand, which is used as a template for the transcription of a gene or cDNA, may be referred to as coding for a protein or other product of that gene or cDNA.

[0103] An "expression vector" refers to a vector containing recombinant polynucleotides that include an expression regulatory sequence operably ligated to the nucleotide sequence to be expressed. An expression vector contains sufficient cis-acting elements for expression; other elements for expression may be supplied by the host cell or in an in vitro expression system. Expression vectors include all vectors known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes), and viruses (e.g., Sendai virus, lentivirus, retrovirus, adenovirus, and adeno-associated virus) that incorporate recombinant polynucleotides.

[0104] As used herein, the term "ex vivo" with respect to cells transfected, transfected, or transformed ex vivo means cells that are transfected, transfected, or transformed outside of the subject, i.e., the cells are removed from the subject before such cells are transfected, transfected, or transformed.

[0105] As used herein, “identity” refers to subunit sequence identity between two polymer molecules, such as between two polynucleotide or polypeptide molecules, or between two nucleic acid or amino acid molecules. Two amino acid sequences are identical if they have the same residue at the same position; for example, if a position in each of two polypeptide molecules is occupied by arginine, they are identical at that position. The degree of identity or identical residue at the same position in alignment between two amino acid or nucleic acid sequences is often expressed as a percentage. Identity between two amino acid or nucleic acid sequences is a linear function of the number of matching or identical positions: for example, if half of the positions in the two sequences are identical, the two sequences are 50% identical, and if 90% of the positions (e.g., 9 out of 10) match or are identical, the two amino acid sequences are 90% identical.

[0106] "Substantially identical" means a polypeptide or nucleic acid molecule that exhibits at least 50% identity with respect to a reference amino acid sequence (e.g., any one of the amino acid sequences described herein) or a nucleic acid sequence (e.g., any one of the nucleic acid sequences described herein). In some embodiments, such sequences are at least 60%, 80%, or 85%, or 90%, 95%, or even 99%, identical at the amino acid level or nucleic acid level to the sequence used for comparison. Other percentages of identity with respect to specific sequences are described herein.

[0107] Sequence identity can be measured / determined using sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX programs). Such software matches identical or similar sequences by assigning a degree of homology to various substitutions, deletions, and / or other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. In an exemplary approach to determining the degree of identity, the BLAST program may be used, and the probability score between e3 and e100 indicates closely related sequences. In some embodiments, sequence identity is determined by using BLAST with default settings.

[0108] To the extent that the embodiments provided herein include compositions comprising various proteins, these proteins may, in some examples, include amino acid sequences having sequence identity with the amino acid sequences disclosed herein. Therefore, in certain embodiments, depending on the particular sequence, the degree of sequence identity is preferably greater than 50% (e.g., 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) with respect to the sequence numbers disclosed herein. In addition to these percentages, other percentages of identity are provided herein. Polypeptide identity can be determined using an affine gap search with parameters of gap-open penalty -12 and gap-extension penalty =1, using the Smith-Waterman homology search algorithm implemented in the MPSRCH program (Oxford Molecular).

[0109] These proteins may contain one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) conserved amino acid substitutions, i.e., substitutions of one amino acid with another amino acid having a corresponding side chain, compared to the disclosed proteins. Genetically encoded amino acids are generally divided into four families: (1) acidic, i.e., aspartates and glutamates; (2) basic, i.e., lysine, arginine, and histidine; (3) nonpolar, i.e., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan; and (4) non-charged, i.e., glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes classified together as aromatic amino acids. In general, single amino acid substitutions within these families do not have a significant impact on biological activity. The protein may have one or more single amino acid deletions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) relative to the disclosed protein sequence. The protein may also contain one or more insertions (e.g., each of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) relative to the disclosed protein sequence (e.g., each of 1, 2, 3, 4, or 5 amino acids).

[0110] As used herein, the term "in vivo" with respect to cells transfected, transfected, or transformed in vivo means cells that are transfected, transfected, or transformed in a subject without the cells being removed from the subject before such cells are transfected, transfected, or transformed.

[0111] "Isolated" means that it has been altered or removed from its natural state. For example, nucleic acids or peptides that are naturally present in living animals are not "isolated," but the same nucleic acids or peptides that have been partially or completely separated from their naturally occurring coexisting substances are "isolated." Isolated nucleic acids or proteins may exist in a substantially purified form or in a non-natural environment, such as a host cell.

[0112] As used herein, "lentivirus" refers to a genus of retroviridae capable of infecting non-dividing cells. Non-exclusive examples of lentiviruses include HIV, SIV, and FIV. Lentivirus-derived vectors or virus-like particles can be used to transduce cells, deliver genes or other molecules, and express them in cells either in vitro (ex vivo) or in vivo.

[0113] As used herein, the term “modified” means an altered state or structure of a molecule or cell provided herein. Molecules can be modified in many ways, including chemical, structural, and functional, such as mutation, substitution, insertion, or deletion (e.g., internal deletion, cleavage). Cells can be modified by the introduction of nucleic acids or the expression of heterologous proteins.

[0114] As used herein, the term “modulate” means mediating an increase or decrease in the level of response in an object compared to the level of response in the object in the absence of the treatment or compound, and / or compared to the level of response in an otherwise identical but untreated object. This term includes disrupting and / or influencing natural signals or responses, thereby mediating a beneficial therapeutic response in an object such as a human.

[0115] Unless otherwise specified, "nucleotide sequences encoding an amino acid sequence" include all nucleotide sequences that encode the same amino acid sequence, including degenerate versions of each other. The phrase "nucleotide sequence encoding a protein or RNA" can also include introns to the extent that a nucleotide sequence encoding a protein may contain introns in some version.

[0116] The term “oligonucleotide” typically refers to a short polynucleotide. When a nucleotide sequence is represented by a DNA sequence (i.e., A, T, C, G), it is understood that this also provides a corresponding RNA sequence (i.e., A, U, C, G) where “U” substitutes for “T”.

[0117] Parenteral administration of the composition includes, for example, subcutaneous (sc), intravenous (iv), intramuscular (im), or intrasternal injection or infusion techniques.

[0118] As used herein, the term “polynucleotide” is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Therefore, as used herein, the terms “nucleic acid” and “polynucleotide” are interchangeable. As used herein, polynucleotide includes, but is not limited to, all nucleic acid sequences obtained by any method available in the art, including recombinant methods, i.e., cloning of nucleic acid sequences from recombinant libraries or cell genomes using cloning techniques and PCR, etc., as well as by synthetic means.

[0119] As used herein, the terms “peptide,” “polypeptide,” and “protein” are interchangeable and refer to compounds consisting of multiple amino acid residues covalently linked by peptide bonds. As used herein, this term refers to both short chains, also commonly referred to in the art as peptides, oligopeptides, and oligomers, and long chains, of which there are many types, commonly referred to in the art as proteins. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, polypeptide variants, modified polypeptides, derivatives, analogs, and fusion proteins. Polypeptides include native peptides, recombinant peptides, synthetic peptides, or combinations thereof.

[0120] Unless otherwise specified, the terms “heteroviral structural protein” and “heteroviral glycoprotein” as used herein are synonymous and interchangeable. Therefore, unless otherwise specified, embodiments referring to “heteroviral structural protein” are understood to refer to “heteroviral glycoprotein,” and vice versa.

[0121] As used herein, the terms “pseudotype” or “pseudotyped viral particle” refer to a viral particle having a glycoprotein derived from another enveloped virus, or a viral vector encoding an envelope glycoprotein from a virus different from the parent virus. Therefore, the host range of the vector particle may be expanded or modified depending on the type of cell surface receptor used by the glycoprotein. For example, a virus may be pseudotyped with the VSV-G mutant protein provided herein.

[0122] As used herein with respect to antibodies, the term “specifically binding” means 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, such interspecies reactivity does not in itself alter the antibody’s classification as specific. In another example, an antibody that specifically binds to an antigen may also bind to different alleles of the antigen. However, such cross-reactivity does not in itself alter the antibody’s classification as specific. In some cases, the term “specific binding” or “specifically binding” may be used in relation to the interaction of an antibody, protein, or peptide with a second chemical species, meaning that the interaction depends on the presence of a specific structure on the chemical species (e.g., an antigenic determinant or epitope); for example, an antibody recognizes and binds to a specific protein structure rather than the protein in general. If an antibody is specific to epitope “A”, the presence of labeled “A” and molecules containing epitope A (or free unlabeled A) in the reactant containing the antibody will reduce the amount of labeled A bound to the antibody. In some embodiments, the targeting moieties described herein, which may be used to target viral particles containing mutant VSV-G proteins or other viral structural proteins used to pseudotype the virus, can specifically bind to those targets.

[0123] The term "subject" includes living organisms, including those capable of eliciting an immune response (e.g., mammals). As used herein, "subject" or "patient" may be human or a non-human mammal. Examples of non-human mammals include livestock and pets such as sheep, cattle, pigs, dogs, non-human primates, cats, and mouse mammals. In some embodiments, the subject is human.

[0124] As used herein, the term “therapeutic” means treatment and / or prevention. Therapeutic effects are obtained by suppression, remission, or eradication of a disease state.

[0125] As used herein, the terms “transfected,” “transformed,” or “transduced” refer to the process by which an exogenous nucleic acid is introduced into or into a cell. A “transfected,” “transformed,” or “transduced” cell is one that has been transfected, transformed, or transduced with an exogenous nucleic acid. Cells include primary target cells and their offspring. In some embodiments, transfection, transformation, or transduction is carried out or occurs in vivo.

[0126] When used herein, “to treat” a disease means to reduce the frequency or severity of at least one sign or symptom of the disease or disorder experienced by the subject.

[0127] A "vector" is a composition of substances containing isolated nucleic acids that encode proteins or peptides. Numerous vectors are known in the art and include, but are 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, retrovirus vectors, and lentiviral vectors.

[0128] The "carrier" or "delivery vehicle" includes viral particles, viruses, polylysine compounds, and liposomes, which facilitate the transfer of nucleic acids into cells. Proteins or peptides can also be delivered to cells using the carrier or delivery vehicle.

[0129] Scope: Throughout this disclosure, various embodiments of the embodiments may be presented in range form. It should be understood that the range form is merely for convenience and brevity and should not be interpreted as an inflexible limitation. Therefore, a range description should be considered to specifically disclose all possible subranges, as well as the individual numbers within that range. For example, a range description such as 1–6 should be considered to specifically disclose subranges such as 1–3, 1–4, 1–5, 2–4, 2–6, 3–6, as well as the individual numbers within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the width of the range. Unless otherwise explicitly stated, the disclosed range includes its endpoints. heterogeneous proteins

[0130] In some embodiments, the polypeptide is a recombinant fusion protein comprising: ii) 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.

[0131] 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 includes an ICD of hGCSF-R, or a variant or fragment thereof. In some embodiments, the hGCSF-R ICD includes substitutions, insertions, or deletions. In some embodiments, the hGCSF-R ICD includes 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 thereto. In some embodiments, the hGCSF-R ICD does not include 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 the amino acid residues 800-813, 775-813, 750-813, 725-813, or 716-813 of SEQ ID NO: 125, or the C-terminal residue of SEQ ID NO: 111.

[0132] In some embodiments, the hGCSF-R ICD includes 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 include the continuous fragment of amino acid residues 716-813 of SEQ ID NO: 125.

[0133] As used herein, the term “ligand-binding extracellular domain (ECD)” refers to a protein domain that can bind to a ligand and act to transmit a signal via the ICD. This signal may enhance cell activation or increase the rate of cell division, thereby increasing the number of cells. The ligand may be naturally occurring, synthetic, or recombinant. For example, G-CSF (granulocyte colony-stimulating factor) or human growth hormone (HGH) can be used as ligands that can bind to the ECD and transmit a signal. For example, somatropin can be used as a ligand that can bind to an ECD that can bind to HGH. Furthermore, in some embodiments, lenograstim (granocyte), filgrastim (newpogen, salzio, nivestim, acofil), long-acting (pegylated) filgrastim (pegylated filgrastim, neurusta, permeg, ziextenko), or lipezilgrastim (Lonquex) can be used as ligands that can bind to G-CSF that can bind to HGH. Examples of ECDs provided herein include, but are not limited to, G-CSFR or hGHR. While not bound by any particular theory, when these drugs are administered to patients expressing these polypeptides, T cells proliferate (increase in number), resulting in a greater response against tumors.

[0134] 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.

[0135] In some embodiments, the ECD includes the ECD of the human growth hormone receptor (hGH-R), or a variant or fragment thereof. In some embodiments, the ECD of the human growth hormone receptor (hGH) includes the amino acid sequence encoded by exons 1-7 of hGH-R, or a variant or fragment thereof. In some embodiments, the ECD of hGH-R includes the C259R variant. In some embodiments, the ECD of hGH-R includes 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 to them. In some embodiments, the ECD of hGH-R includes 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 to it.

[0136] 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 the 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 the gp130 ICD, or a fragment thereof, is the amino acid sequence of the gp130 ICD of construct G, which is described in more detail herein.

[0137] In some embodiments, the ICD includes a fragment of GCSF-R ICD. In some embodiments, the GCSF-R ICD fragment does not include the STAT3, SH3, and / or SHC domains. In some embodiments, the GCSF-R ICD fragment includes a Jak1 and / or STAT3 binding site. In some embodiments, the ICD includes IL-2Rβ ICD, or a fragment or variant thereof. In some embodiments, IL-2Rβ ICD is a fragment of IL-2Rβ ICD. In some embodiments, IL-2Rβ ICD, or a fragment thereof, includes an SHC or STAT5 binding domain. In some embodiments, the ICD includes IL-7Rα ICD, or a fragment or variant thereof. In some embodiments, IL-7RαICD is a fragment of IL-7RαICD. In some embodiments, the ICD, or a fragment thereof, includes a STAT5 and / or PI3K binding domain. In some embodiments, the ICD includes an IL-2Rβ ICD, or a gp130 ICD or a variant or fragment thereof linked to an IL-2Rβ ICD. An unrestricted example of an IL-2Rβ ICD, or a fragment thereof, is the amino acid sequence of SEQ ID NO: 123. In some embodiments, the ICD includes an IL-7Rα ICD, or a hCCSF-R ICD, or a variant or fragment thereof linked to an IL-7Rα ICD. An unrestricted example of an IL-7Rα ICD, or a fragment thereof, is the amino acid sequence of SEQ ID NO: 118. Another unrestricted example of an IL-7Rα ICD, or a fragment thereof, is the amino acid sequence of the IL-7Rα ICD of construct E, which is described in more detail herein.

[0138] In some embodiments, the transmembrane domain includes a GCSF-R, gp130, or hGH-R transmembrane domain. In some embodiments, the transmembrane domain includes an hGCSF-R transmembrane domain or a variant thereof. In some embodiments, the transmembrane domain includes a gp130 transmembrane domain or a variant thereof. An example of a gp130 transmembrane domain is the amino acid sequence of SEQ ID NO: 121, but is not limited thereto. In some embodiments, the transmembrane domain includes an hGH-R transmembrane domain or a variant thereof.

[0139] In some embodiments, the polypeptide comprises an ECD which is a GCSFR-ECD, or a variant thereof (e.g., as described herein and above); a transmembrane domain which is a GCSF-R transmembrane domain, or a variant thereof (e.g., as described herein and above); and an immunoactivated ICD which is a GCSF-R ICD, or a variant thereof (e.g., as described herein and above).

[0140] In some embodiments, the polypeptide comprises an ECD which is hGH-R-ECD, or a variant thereof (e.g., as described herein and above); a transmembrane domain which is a GCSF-R transmembrane domain, or a variant thereof (e.g., as described herein and above); and an immunoactivated ICD which is a GCSF-R ICD, or a variant thereof (e.g., as described herein and above).

[0141] In some embodiments, the polypeptide comprises an ECD which is hGH-R-ECD, or a variant thereof (e.g., as described herein and above); a transmembrane domain which is a gp130 transmembrane domain, or a variant thereof (e.g., as described herein and above); and an immunoactivated ICD containing a gp130 ICD, or a variant thereof (e.g., as described herein) and an IL-2Rβ ICD, or a variant thereof (e.g., as described herein and above).

[0142] In some embodiments, the polypeptide comprises an ECD which is hGH-R-ECD, or a variant thereof (e.g., as described herein and above); a transmembrane domain which is a GCSF-R transmembrane domain, or a variant thereof (e.g., as described herein and above); and an immunoactivated ICD which includes a GCSF-R ICD, or a variant thereof (e.g., as described herein and above); and an IL-7Rα ICD, or a variant thereof (e.g., as described herein and above).

[0143] 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 any of the aforementioned variants. 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 respect to 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.

[0144] Examples of these domains or sequences are provided herein.

[0145] Polypeptides having different domains described in this section are distinct from those known as chimeric antigen receptors. While chimeric antigen receptors may reside in the same cells as these polypeptides, these polypeptides would not be considered chimeric antigen receptors by those skilled in the art. Therefore, in some embodiments, the polypeptides do not contain an antigen-binding domain. For example, in some embodiments, the polypeptides do not contain an antigen-binding domain that is scFv, Fab, or in the form of a single-domain antibody. Other antigen-binding domains are provided herein, and in some embodiments, the polypeptides do not contain any form of antigen-binding domain. Those skilled in the art would not consider a ligand ECD domain to be the same as an antigen-binding domain. Examples of domains that polypeptides 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.

[0146] In some embodiments, these polypeptides do not contain an ECD containing a binding domain that binds to a tumor antigen. In some embodiments, the polypeptides do not contain a costimulatory domain. In some embodiments, the polypeptides do not contain a CD3ζ signaling domain. In some embodiments, the polypeptides do not contain a CD28ζ signaling domain. In some embodiments, the polypeptides do not contain an immune receptor tyrosine-based activation motif (ITAM). In some embodiments, the polypeptides do not contain a costimulatory endodomain. In some embodiments, the polypeptides do not contain a 4-1BB signaling or costimulatory endodomain. In some embodiments, the polypeptides do not contain a CD28 signaling or costimulatory endodomain. In some embodiments, the polypeptides do not contain an inducible T cell costimulatory (ICOS) intracellular domain.

[0147] In some embodiments, the polypeptide does not contain the CH2, CH3, CD28, or CD8α hinge region.

[0148] In some embodiments, polypeptides containing the ECD of G-CSFR or their variants, such as those provided herein, do not contain an antigen-binding domain.

[0149] These polypeptides can be incorporated into any of the particles provided herein. According to some embodiments, particles comprising polypeptides are provided. 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 vectors are pseudotyped viral vectors. Non-limiting examples of pseudotyped viral vectors are provided herein, but they can be pseudotyped with any viral structural protein. In some embodiments, the particles further comprise chimeric antigen receptors, for example, chimeric antigen receptors comprising an antigen-binding domain that binds to a tumor antigen.

[0150] Nucleic acid molecules encoding chimeric antigen receptors (CARs), polypeptides, or combinations thereof are also provided herein. In some embodiments, the CAR is as provided herein. In some embodiments, the CAR and polypeptide are encoded by a unique nucleic acid molecule. In some embodiments, the CAR and polypeptide are encoded by the same nucleic acid molecule. Methods for encoding multiple proteins of interest from a single nucleic acid molecule are known in the art, and any such method is within the scope of this disclosure. In some embodiments, a single nucleic acid molecule encodes the CAR and polypeptide as a bisistronic molecule, and the CAR and polypeptide 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 bisistronic construct, where, from 5' to 3', the nucleic acid molecule encodes the CAR, a cleavable linker (e.g., a 2A linker), and a polypeptide. In some embodiments, the nucleic acid molecule encodes a bisistronic construct, and from 5' to 3', the nucleic acid molecule encodes a polypeptide, a cleavable linker (e.g., a 2A linker), and a CAR. In some embodiments, particles comprising the nucleic acid molecule are provided. 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 vectors are pseudotyped viral vectors. Non-limiting examples of pseudotyped viral vectors are provided herein, but any viral structural protein can be pseudotyped. Viral particles containing heterologous viral glycoproteins and targeting moieties

[0151] In some embodiments, a viral particle comprising a heterologous viral glycoprotein and a targeting moiety is provided. In some embodiments, the targeting moiety comprises a polypeptide having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. In some embodiments, S1 comprises a modified Fc protein, which comprises a transmembrane domain such as, but not limited to, a CD8 or CD28 transmembrane domain. Thus, in some embodiments, the stalk moiety S1 comprises an N-terminal to C-terminal orientation of the modified Fc transmembrane domain. In some embodiments, the modified Fc protein comprises an effector mutation, which inhibits the interaction between the Fc protein and Fc-interacting proteins such as FcγR, C1q, FcRβ, or FcRn.

[0152] In some embodiments, the S1 stalk portion is bound to the surface of the virus particle via a transmembrane domain. In some embodiments, the modified Fc protein is a variant of the IgG1 Fc, IgG2 Fc, or IgG4 Fc protein. In some embodiments, the modified Fc protein includes a variant of the sequence of SEQ ID NO: 26 (IgG1 Fc), SEQ ID NO: 27 (IgG2 Fc), or SEQ ID NO: 28 (IgG4 Fc).

[0153] In some embodiments, the modified Fc protein is the modified IgG1 Fc protein (SEQ ID NO: 26). In some embodiments, the modified IgG1 Fc protein contains one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A of SEQ ID NO: 26, numbered according to Kabat's EU numbering index, as described in Edelman, GM et al., "The covalent structure of an entire 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, the whole of which is incorporated herein by reference. Any of the mutations L234A, L235A, N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 26 may or may not be present, and the mutations may be combined in any combination. In some embodiments, the modified IgG1 Fc protein contains mutations corresponding to L234A and L235A in SEQ ID NO: 26. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to N297A in SEQ ID NO: 26. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to P329G in SEQ ID NO: 26. In some embodiments, the modified IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, and P329G in SEQ ID NO: 26. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to I253A in SEQ ID NO: 26. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to H310A in SEQ ID NO: 26. In some embodiments, the modified IgG1 Fc protein contains the mutation corresponding to H435A in SEQ ID NO: 26.In some embodiments, the modified IgG1 Fc protein contains mutations corresponding to I253A, H310A, and H435A in SEQ ID NO: 26. In some embodiments, the modified IgG1 Fc protein contains mutations corresponding to L234A, L235A, N297A, P329G, I253A, H310A, and H435A in SEQ ID NO: 26.

[0154] In some embodiments, the modified Fc protein, including the modified IgG1 Fc protein, includes a cleavage of the IgG1 Fc sequence. The cleavage 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 modified Fc protein, including the modified IgG1 Fc protein, includes a cleavage of SEQ ID NO: 26. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the SEQ ID NO: 26. In some embodiments, the cleavage includes the deletion of an amino acid from the N-terminus of SEQ ID NO: 26. In some embodiments, the cleavage includes the deletion of an amino acid from the C-terminus of SEQ ID NO: 26. In some embodiments, the cleavage includes the deletion of an amino acid from both the N-terminus and C-terminus of SEQ ID NO: 26. In some embodiments, the cleavage of SEQ ID NO: 26 includes the amino acid sequence of SEQ ID NO: 103. EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(Sequence ID 103)

[0155] In some embodiments, the modified IgG1 Fc protein contains one or more mutations selected from the group consisting of L19A, L20A, N82A, P114G, I38A, H95A, and H220A, compared to SEQ ID NO: 103. 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. Any of 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 modified IgG1 Fc protein contains mutations corresponding to L19A and L20A of SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to N82A of SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to P114G of SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains mutations corresponding to L19A, L20A, N82A, and P114G of SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to I38A of SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains a mutation corresponding to H95A of SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains the mutation corresponding to H220A in SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains the mutations corresponding to I38A, H95A, and H220A in SEQ ID NO: 103. In some embodiments, the modified IgG1 Fc protein contains the mutations corresponding to L19A, L20A, N82A, P114G, I38A, H95A, and H220A in SEQ ID NO: 103.In some embodiments, the modified IgG1 Fc protein contains mutations corresponding to L18A, L19A, N82A, P114G, I38A, H95A, and H220A, respectively, in SEQ ID NO: 103. Exemplary modified IgG1 proteins containing L18A, L19A, N82A, P114G, I38A, H95A, and H220A are shown below in the amino acid sequence of SEQ ID NO: 104: EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMASRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLAQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNAYTQKSLSLSPGK(Sequence ID 104)

[0156] In some embodiments, the modified Fc protein is the modified IgG2 Fc protein (SEQ ID NO: 27). In some embodiments, the modified IgG2 Fc protein contains one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A, where these positions correspond to SEQ ID NO: 27 numbered according to Kabat's EU numbering index. 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 modified IgG2 Fc protein contains the mutation corresponding to N297A in SEQ ID NO: 27. In some embodiments, the modified IgG2 Fc protein contains the mutation corresponding to P329G in SEQ ID NO: 27. In some embodiments, the modified IgG2 Fc protein contains the mutations corresponding to N297A and P329G in SEQ ID NO: 27. In some embodiments, the modified IgG2 Fc protein contains a mutation corresponding to I253A in SEQ ID NO: 27. In some embodiments, the modified IgG2 Fc protein contains a mutation corresponding to H310A in SEQ ID NO: 27. In some embodiments, the modified IgG2 Fc protein contains a mutation corresponding to H435A in SEQ ID NO: 27. In some embodiments, the modified IgG2 Fc protein contains mutations corresponding to I253A, H310A, and H435A in SEQ ID NO: 27.

[0157] In some embodiments, the modified Fc protein, including the modified IgG2 Fc protein, includes a cleavage of the IgG2 Fc sequence. The cleavage 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 modified Fc protein, including the modified IgG2 Fc protein, includes a cleavage of SEQ ID NO: 27. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the SEQ ID NO: 27. In some embodiments, the cleavage includes the deletion of an amino acid from the N-terminus of SEQ ID NO: 27. In some embodiments, the cleavage includes the deletion of an amino acid from the C-terminus of SEQ ID NO: 27. In some embodiments, the cleavage includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 27.

[0158] In some embodiments, the modified Fc protein is the modified IgG4 Fc protein (SEQ ID NO: 28). In some embodiments, the modified IgG4 Fc protein contains one or more mutations selected from the group consisting of S228P, L235E, N297A, P329G, I253A, H310A, and H435A, where these positions correspond to SEQ ID NO: 28 numbered according to Kabat's EU numbering index. 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 modified IgG4 Fc protein contains the mutation corresponding to S228P in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains the mutation corresponding to L235E in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains a mutation corresponding to N297A in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains a mutation corresponding to P329G in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains mutations corresponding to S228P, L235E, N297A, and P329G in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains a mutation corresponding to I253A in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains a mutation corresponding to H310A in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains a mutation corresponding to H435A in SEQ ID NO: 28. In some embodiments, the modified IgG4 Fc protein contains mutations corresponding to I253A, H310A, and H435A in SEQ ID NO: 28.

[0159] In some embodiments, the modified Fc protein, including the modified IgG4 Fc protein, includes a cleavage of the IgG4 Fc sequence. The cleavage 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 modified Fc protein, including the modified IgG4 Fc protein, includes a cleavage of SEQ ID NO: 28. The cleavage may include the deletion of any number of amino acids from the N-terminus, C-terminus, or both of the SEQ ID NO: 28. In some embodiments, the cleavage includes the deletion of an amino acid from the N-terminus of SEQ ID NO: 28. In some embodiments, the cleavage includes the deletion of an amino acid from the C-terminus of SEQ ID NO: 28. In some embodiments, the cleavage includes the deletion of amino acids from both the N-terminus and C-terminus of SEQ ID NO: 28.

[0160] In some embodiments, the stalk moiety S1, comprising the modified Fc protein, is given by the formula L1-Fc-L2-X1, where L1 is a linker or absent; Fc is the modified Fc protein; L2 is a linker or absent; and X1 is a polypeptide comprising a transmembrane domain. Thus, the targeting moiety comprising the formula T-S1 may also be given by the formula T-L1-Fc-L2-X1, where T is a target-binding domain; L1 is a linker or absent; Fc is the modified Fc protein; L2 is a linker or absent; and X1 is a polypeptide comprising a transmembrane domain. Therefore, it should be understood that in some embodiments, the stalk moiety S1 may 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 modified Fc protein is as provided herein.

[0161] In some embodiments, L1 and L2 are each independently polypeptide linkers. In some embodiments, the polypeptide linker is (GGGGA) n (Sequence No. 54), (GGGGS) n (Sequence ID 55), (EAAAK) n(Sequence ID 73), A(EAAAK) n A (Sequence ID 74), (XP) n The formula includes (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 a combination thereof, where each n is independently 1 to 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 absent. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55) , In the formula, each n is independently 1 to 5. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 1. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 2. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55) , And in the formula, each n is independently 3. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 4. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n(Sequence ID 55), where each n is independently 5. In some embodiments, L1 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55) , In the formula, each n is independently greater than 5. In some embodiments, L2 does not exist. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55) is given by the formula, where each n is independently 1 to 5. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55) , And each n is independently 1. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 2. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 3. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 4. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence ID 55), where each n is independently 5. In some embodiments, L2 is (GGGGA) n (Sequence No. 54) or (GGGGS) n (Sequence code 55), where each n is independently greater than 5.

[0162] In some embodiments, X1 is ECD-T M - Contains a polypeptide having the formula ICD, where ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent; TM is the transmembrane domain of a transmembrane protein; ICD is either the intracellular domain of the protein, or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or is absent. Therefore, in some embodiments, the targeting portion comprising formula T-S1, which can also be given by formula T-L1-Fc-L2-X1, is formula T-L1-Fc-L2-ECD-T M -Can also be given by ICD, where T is the target-binding domain; L1 is a linker or is absent; Fc is a modified Fc protein; L2 is a linker or is absent; ECD is the extracellular domain of a cell surface protein or a fragment thereof or is absent; T M is the transmembrane domain of a transmembrane protein; ICD is either the intracellular domain of the protein, or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or is absent. Therefore, in some embodiments, the stalk portion S1 is of the formula L1-Fc-L2-ECD-T M -Please understand what can be provided by ICD.

[0163] In some embodiments, the stalk portion S1 does not include the modified Fc region. In some embodiments, the stalk portion S1 is given by formula L3-X1, where L3 is a flexible peptide linker and X1 is a polypeptide comprising a transmembrane domain provided herein. Thus, in some embodiments, the targeting portion comprising 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 comprising a transmembrane domain provided herein. Thus, it should be understood that in some embodiments, the stalk portion S1 may be given by formula L3-X-1. In some embodiments, the S1 stalk portion is bound to the surface of the virus particle via a transmembrane domain.

[0164] In some embodiments, the flexible peptide linker L3 may be any flexible peptide linker. In some embodiments, L3 may be, but is not limited to (GGGGA) n (Sequence No. 54), (GGGGS) n A flexible linker is selected from the group including (SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESKST (SEQ ID NO: 58), or any combination thereof, where each n is an integer independently selected from 1 to 4. In some embodiments, each n is an integer independently 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 (Sequence ID 54) where n is 1. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 2. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 3. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 4. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 5. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 6. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 7. In some embodiments, L3 is (GGGGA)n (SEQ ID NO: 54), and n is 8. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54), and n is 9. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54), and n is 10. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54), and n is greater than 10. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 1. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 2. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 3. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 4. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 5. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 6. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 7. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 8. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 9. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55), and n is 10. In some embodiments, 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).

[0165] In some embodiments, the flexible peptide linker L3 can be any flexible peptide linker. In some embodiments, L3 includes, but is 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), EGKSSGSGSESKST (SEQ ID NO: 58), or any combination thereof, and is selected from the group of flexible linkers, where 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 embodiments, L3 is (GGGGA) n (SEQ ID NO: 54) and n is 2. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54) and n is 3. In some embodiments, L3 is (GGGGA) n (SEQ ID NO: 54) and n is 4. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55) and n is 1. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55) and n is 2. In some embodiments, L3 is (GGGGS) n (SEQ ID NO: 55) and n is 3. In some embodiments, 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).

[0166] In some embodiments, L3 is, but is not limited to (GGGGA) n (Sequence No. 54), (GGGGS) n A flexible linker is selected from the group including (SEQ ID NO: 55), GSAGSAAGSGEF (SEQ ID NO: 56), KESGSVSSEQLAQFRSLD (SEQ ID NO: 57), EGKSSGSGSESKST (SEQ ID NO: 58), or any combination thereof, where each n is an integer independently 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 (Sequence ID 54) where n is 1. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 2. In some embodiments, L3 is (GGGGA) n (Sequence ID 54) where n is 4. In some embodiments, L3 is (GGGGS) n (Sequence ID 55), where n is 1. In some embodiments, L3 is (GGGGS) n (Sequence ID 55), where n is 2. In some embodiments, L3 is (GGGGS) n (Sequence ID 55) where n is 4. In some embodiments, L3 is GSAGSAAGSGEF (Sequence ID 56). In some embodiments, L3 is KESGSVSSEQLAQFRSLD (Sequence ID 57). In some embodiments, L3 is EGKSSGSGSESKST (Sequence ID 58).

[0167] In some embodiments, X1 is ECD-T M - Contains a polypeptide having the formula ICD, where ECD is either the extracellular domain of a cell surface protein or a fragment thereof, or is absent; T Mis the transmembrane domain of a transmembrane protein; ICD is either the intracellular domain of the protein, or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or is absent. Therefore, in some embodiments, the targeting portion comprising formula T-S1, which can also be given by formula T-L3-X1, is formula T-L3-ECD-T M -Can also be given by ICD, 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 is absent; T M is the transmembrane domain of a transmembrane protein; ICD is either the intracellular domain of the protein, or a protein that facilitates the incorporation of the targeting portion into the envelope of a viral particle, or is absent. Therefore, in some embodiments, the stroke portion S1 is of formula L3-ECD-T M -Please understand what can be provided by ICD.

[0168] In some embodiments, the ECD is absent. In some embodiments, the ECD may be any suitable extracellular domain or a fragment thereof. In some embodiments, the ECD may originate from a different protein compared to the transmembrane domain. The ECD domain may 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 contains the polypeptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the CD8 ECD domain consists of or is essentially the polypeptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain contains a polypeptide that is 25 to 45 amino acids long. In some embodiments, the ECD comprises a polypeptide that is at least or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, the ECD domain is the CD28 ECD domain or a fragment thereof. In some embodiments, the CD28 ECD domain comprises the polypeptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the CD28 ECD domain consists of or is essentially the polypeptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, the ECD domain comprises a polypeptide that is 25 to 45 amino acids long.In some embodiments, the ECD comprises a polypeptide that is at least or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60).

[0169] In some embodiments, T M This can be any suitable transmembrane domain or fragment thereof. In some embodiments, T M The domain is CD8 or CD28T. M It is a domain or a fragment thereof. In some embodiments, T M The domain is CD8 T M It is a domain or a fragment thereof. In some embodiments, CD8 T M The domain contains the polypeptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, CD8 T M The domain consists of, or is essentially, the polypeptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M This comprises a polypeptide that is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide of IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61). In some embodiments, T M The domain is CD28 T M It is a domain or a fragment thereof. In some embodiments, CD28 T M The domain contains the polypeptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, CD28 T M The domain consists of, or is essentially, the polypeptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62). In some embodiments, T MThis includes a polypeptide that is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62).

[0170] In some embodiments, T M The domain originates from the same protein as ECD. In some embodiments, T M The domain originates from a protein different from ECD. In some embodiments, T M The domain is CD8 or CD28T. M A domain or fragment thereof, and an ECD domain is a CD8 or CD28 ECD domain or fragment thereof. In some embodiments, T M The domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide IYIWAPLAGTCGVLLLSLVITLYCNHRN (SEQ ID NO: 61), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60). In some embodiments, T MThe domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 59). In some embodiments, T M The domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 62), and the ECD domain is at least, or about 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the peptide KIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP (SEQ ID NO: 60).

[0171] In some embodiments, the transmembrane domain is linked to the intracellular domain (ICD) of a transmembrane protein, or a fragment thereof. In some embodiments, the ICD is absent. In some embodiments, the ICD is derived from the same or a different protein as the TM domain. In some embodiments, the ICD includes an Env integration motif. The Env integration motif is a molecule, e.g., a polypeptide, that may help facilitate the integration of proteins into the viral envelope. A non-limiting example of an Env integration motif is a polypeptide containing the amino acid sequence NRVRQGYS (SEQ ID NO: 63). This is a non-limiting example, and other peptide sequences, e.g., GGTETSQVAPA (SEQ ID NO: 64), may be used. In some embodiments, the Env integration motif includes the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 64, or a combination thereof. In some embodiments, the Env integration motif includes the amino acid sequence of SEQ ID NO: 63. In some embodiments, the Env integration motif includes the amino acid sequence of SEQ ID NO: 64.

[0172] In some embodiments, the target-binding domain "T" is any polypeptide or polynucleotide that can be used to bind to a desired target. In some embodiments, T is one of the following: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR alpha, TCR beta, TCR gamma, TCR delta, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3; a glycosylated CD43 epitope expressed in acute leukemia or lymphoma but not in hematopoietic progenitor cells; a glycosylated CD43 epitope expressed on non-hematopoietic cancers; A kinase anchor protein 4 (AKAP-4); adrenaline receptor β3 (ADRB3); AFP; anaplastic lymphoma kinase kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie2); autoantibody against desmoglein 1 (Dsg1); autoantibody against desmoglein 3 (Dsg3); B7H3 (CD276); biotin; bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; cancer / testicular antigen 1 (NY-ESO-1); cancer / testicular antigen 2 (LAGE-LA); carbonic anhydrase IX (CA1X); carcinoembryonic antigen (CEA); CC CTC binding factor (Zinc Finger Protein)-like (BORIS or Brother of the Regulator of lmprinted Sites); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32(FCGR2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGLll; CD200R; CD276 / B7H3; CD300 molecule-like family member f (CD300LF);CDH1-CD324; CDH6; CDH17; CDH19; chromosome X open reading frame 61 (CXORF61); Claudin 6 (CLDN6); Claudin 18.2 (CLD18A2 or CLDN18A.2); CMV pp65; C-MYC epitope tag; Cripto; CS1 (also called 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-containing mucin-like hormone receptor 2 (EMR2);elongation factor 2 mutated (ELF2M);ephrin B2;ephrin type A receptor 2 (EphA2);epidermal growth factor receptor (EGFR);epidermal growth factor receptor variant III (EGFRviii);epidermal cell adhesion molecule (EPCAM);ERG;ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML);IgA receptor Fc fragment (FCAR or CD89);Fc receptor-like 5 5, FCRL5); Fibroblast-activating protein α (FAP); FITC; Fms-like tyrosine kinase 3 (FLT3); Folate receptor alpha (FRa or FR1); Folate receptor β (FRb); Follicle-stimulating hormone receptor (FSHR); Fos-related antigen 1; Fucosyl-GM1; G protein-coupled receptor class C group 5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; Ganglioside G2 (GD2); Ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(ll)Cer);Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l-4)bDGlcp(ll)Cer); GD3; GFRalpha4; Glycoprotein 100 (gplOO); Glypican-3 (GPC3); Gonadotropin hormone receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylyl cyclase C (GCC); Heat shock protein 70-2 mutated (mut hsp70-2); Hepatitis A virus cellular receptor 1 1. HAVCR1; hexasaccharide portion of globoH glycoceramide (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 papilloma virus E6 (HPV E6); human papilloma virus E7 (HPV E7); human telomerase reverse transcriptase (hTERT); IgE; IL13Ra2; IL1 lRa; immunoglobulin λ-like polypeptide 1 (IGLL1); influenza A hemagglutinin (HA); insulin-like growth factor 1 receptor (IGF-I receptor); interleukin-11 receptor alpha (IL-llRa); interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2); intestinal carboxylesterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; Regmine; 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; Gene locus K9 (LY6K); Low conductance chloride channel; Lymphocyte antigen 6 complex; Lymphocyte antigen 75 (LY75); Lymphocyte-specific protein tyrosine kinase (LCK); Mammary gland differentiation antigen (NY-BR-1); Melanoma antigen recognized by T cell 1 (Melanoa or MART 1); Melanoma-associated antigen 1 (MAGE-A1); Melanoma cancer testis antigen-1 antigen-1, MAD-CT-1); melanoma cancer testis antigen-2 (MAD-CT-2); melanoma inhibitor of apoptosis (ML-IAP); mesothelin; MPL; mucin 1 cell surface-bound (MUC1); N-acetylglucosaminyltransferase V (NA17); nectin-4; neural cell adhesion molecule (NCAM); NKG2D; NYBR1; O-acetyl-GD2 ganglioside (OAcGD2); olfactory receptor 51E2 (OR51E2); breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1. 1. Oncogene fusion protein consisting of Ab1 (bcr-ab1); P53 variant; paired box protein Pax-3 (PAX3); paired box protein Pax-5 (PAX5); pannexin 3 (PANX3); PDL1; P-glycoprotein;Placenta-specific 1 (PLAC1); Platelet-derived growth factor receptor beta (PDGFR-beta); Polysialic acid; Proacrosin-binding protein sp32 (OY-TES1); Prostase; Prostate cancer tumor antigen-1 (PCT A-1 or galectin 8); Prostate stem cell antigen (PSCA); Prostate-specific membrane antigen (PSMA); Prostatic acid phosphatase (PAP); Prostain; Protease serine 21 (Testisin or PRSS21); Proteasome (Prosome, Macropain) subunit beta type 9 (LMP2); PTK7; Ras G12V; Ras homolog family member C (RhoC); Rat sarcoma (Rat sarcoma (Ras) variant; advanced glycation end product receptor (RAGE-1); receptor tyrosine kinase-like orphan receptor 1 (ROR1); receptor tyrosine-protein kinase ERBB2 or Her-22 / neu; renal ubiquitous 1 (renal ubiquitous 2, RU1); renal ubiquitous 2 (renal ubiquitous 2, RU2); sarcoma translocation breakpoint; serine 2 (TMPRSS2) ETS fusion gene; sialyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialyl Lewis antigen); sperm protein 17 (SPA17); squamous cell carcinoma antigen 3 (SART3) recognized by T cells; stage-specific embryonic antigen-4 (SSEA-4); STEAP1; Survivin; synovial sarcoma, X breakpoint 2. SSX2); TCR-γ Alternate Reading Frame Protein (TARP); TCR-beta-1 chain; TCR-beta-2 chain; TCR-delta chain; TCR-gamma chain; TCR-gamma-delta chain;Telomerase; TGFbetaR2; 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-related (TEM7R); Tumor protein p53 (p53); Tumor-associated glycoprotein 72 (TAG72); Tyrosinase; Tyrosinase-related protein 2 (TRP-2); Uroplakin 2 (uroplakin 2. UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein (WT1);Alternatively, it is any polypeptide, polynucleotide, or fragment thereof that binds to the X antigen family, member 1A (XAGE1). In some embodiments, the targeting moiety "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. In the context of this disclosure, it should be understood that "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 antibody fragments of any isotype, including but not limited to Fab, Fv, scFv, and Fd fragments, chimeric antibodies, humanized antibodies, single-chain antibodies (scAb), single-domain antibodies (dAb), single-domain heavy-chain antibodies, single-domain light-chain antibodies, bispecific antibodies, multispecific antibodies, and fusion proteins comprising the antigen-binding moiety of an antibody and a non-antibody protein. In some embodiments, the antibody is selected from the group including scFv, Fab, VHH, single-domain antibodies, etc. In some embodiments, the antibody is scFv. In some embodiments, the antibody is Fab. In some embodiments, the antibody is a VHH antibody. In some embodiments, the antibody is a single-domain antibody.

[0173] In some embodiments, the viral particles provided herein are pseudotyped viral particles. In some embodiments, the viral particles are pseudotyped using viral glycoproteins of Paramyxoviridae viruses. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Morbilliviruses, such as measles virus. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Measles virus. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Henipaviruses, such as Nipah virus, Cedar virus, or Hendra virus. In some embodiments, the pseudotyped virus-like particles are pseudotyped using viral glycoproteins of Nipah virus. In some embodiments, the polypeptides or antibodies provided herein are linked to envelope glycoproteins G or H of Paramyxoviridae viruses via a linker. In some embodiments, the Paramyxoviridae viruses are Morbilliviruses, such as measles virus. In some embodiments, the Paramyxoviridae viruses are Henipaviruses, such as Nipah virus, Cedar virus, or Hendra virus.

[0174] As provided herein, viruses can be pseudotyped with the VSV-G protein (either its wild-type or a mutant). While not bound by any particular theory, a mutant VSV-G protein containing a mutation at position 182 (e.g., a lentivirus) can be used to pseudotype a virus and transduce cells if the virus contains a targeting moiety. This mutation inhibits or reduces the affinity of VSV-G for its native 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 the targeted cells.

[0175] In some embodiments, VSV-G proteins are provided that contain a mutation at position 198 compared to SEQ ID NO: 1, or at position 182 compared to SEQ ID NO: 2. SEQ ID NO: 1 is the full-length protein, and SEQ ID NO: 2 is the ectodomain of the VSV-G protein. The 16-mer signal peptide 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. Thus, the mutation can be mentioned in the context of SEQ ID NO: 2, but it should be understood that it is also made in the context of SEQ ID NO: 1, which includes the leader sequence, and is therefore 16 position numbers greater than the position enumerated for 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 embodiments, the mutation is the I182E mutation compared to SEQ ID NO: 2.

[0176] In some embodiments, VSV-G proteins are provided that contain a mutation at position 198 compared to SEQ ID NO: 10, or at position 182 compared to SEQ ID NO: 11. SEQ ID NO: 10 is the full-length protein, and SEQ ID NO: 11 is the ectodomain of the VSV-G protein. The 16-mer signal peptide 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, it should be understood that the mutation may be referred to in the context of SEQ ID NO: 11, but also in the context of SEQ ID NO: 10, which includes the leader sequence, and is therefore 16 position numbers greater than the position enumerated for 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 embodiments, the mutation is the T182E mutation compared to SEQ ID NO: 11.

[0177] In some embodiments, VSV-G proteins are provided that contain a mutation at position 198 compared to SEQ ID NO: 12, or at position 182 compared to SEQ ID NO: 13. SEQ ID NO: 12 is the full-length protein, and SEQ ID NO: 13 is the ectodomain of the VSV-G protein. The 16-mer signal peptide MLRLFLFCFLALGAHS (SEQ ID NO: 67), as shown at the N-terminus of SEQ ID NO: 12, is cleaved, leaving the protein of SEQ ID NO: 13. Therefore, it should be understood that the mutation may be referred to in the context of SEQ ID NO: 13, but also in the context of SEQ ID NO: 12, which includes the leader sequence, and is therefore 16 position numbers greater than the position enumerated for 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 embodiments, the mutation is the A182E mutation compared to SEQ ID NO: 13.

[0178] In some embodiments, VSV-G proteins are provided that contain a mutation at position 203 compared to SEQ ID NO: 14, or at position 182 compared to SEQ ID NO: 15. SEQ ID NO: 14 is the full-length protein, and SEQ ID NO: 15 is the ectodomain of the VSV-G protein. The 21-mer signal peptide MKMKMVIAGLILCIGILPAIG (SEQ ID NO: 68) shown at the N-terminus of SEQ ID NO: 14 is cleaved, leaving the protein of SEQ ID NO: 15. Therefore, it should be understood that the mutation may be referred to in the context of SEQ ID NO: 15, but also in the context of SEQ ID NO: 14, which includes the leader sequence, and is therefore 21 positions higher than the position listed for 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 embodiments, the mutation is the V182E mutation compared to SEQ ID NO: 15.

[0179] In some embodiments, VSV-G proteins are provided that contain a mutation at position 199 compared to SEQ ID NO: 16, or at position 182 compared to SEQ ID NO: 17. SEQ ID NO: 16 is the full-length protein, and SEQ ID NO: 17 is the ectodomain of the VSV-G protein. The 17-mer signal peptide MTPAFILCMLLAGSSWA (SEQ ID NO: 69) shown at the N-terminus of SEQ ID NO: 16 is cleaved, leaving the protein of SEQ ID NO: 17. Therefore, the mutation may be referred to in the context of SEQ ID NO: 17, but it should be understood that it may also be referred to in the context of SEQ ID NO: 16, which includes the leader sequence, and therefore has a position number 17 higher than the position listed for 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 embodiments, the mutation is the V182E mutation compared to SEQ ID NO: 17.

[0180] In some embodiments, VSV-G proteins are provided that contain a mutation at position 199 compared to SEQ ID NO: 18, or at position 182 compared to SEQ ID NO: 19. SEQ ID NO: 18 is the full-length protein, and SEQ ID NO: 19 is the ectodomain 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, it should be understood that the mutation may be referred to in the context of SEQ ID NO: 19, but also in the context of SEQ ID NO: 18, which includes the leader sequence, and is therefore 17 positions higher than the position enumerated with respect to 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 embodiments, the mutation is the V182E mutation compared to SEQ ID NO: 19.

[0181] In some embodiments, VSV-G proteins are provided that contain a mutation at position 199 compared to SEQ ID NO: 20, or at position 182 compared to SEQ ID NO: 21. SEQ ID NO: 20 is the full-length protein, and SEQ ID NO: 21 is the ectodomain of the VSV-G protein. The 17-mer signal peptide MLVLYLLLSLLALGAQC (SEQ ID NO: 71) shown at the N-terminus of SEQ ID NO: 20 is cleaved, leaving the protein of SEQ ID NO: 21. Therefore, it should be understood that the mutation may be referred to in the context of SEQ ID NO: 21, but also in the context of SEQ ID NO: 20, which includes the leader sequence, and is therefore 17 position numbers greater than the position listed for 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 embodiments, the mutation is the I182E mutation compared to SEQ ID NO: 21.

[0182] Where used herein, when a polypeptide is said to have mutations compared to a reference sequence, such comparison is based on alignment, such as using BlastP, ClustalW, or ClutalOmega alignment software with default parameters. For example, position 182 can be found in SEQ ID NO: 2 and can also be found when comparing with other strains, as shown in Figure 3. Figure 3 shows cluster alignments of wild-type sequences of the external domains of various strains of the VSV-G protein. The residues in bold and underlined are the residues that align to position 182 of SEQ ID NO: 2 in various strains. SEQ ID NO: 2 refers to the ectodomain of the VSV-G protein from the Indiana strain. SEQ ID NO: 11 refers to the ectodomain of the VSV-G protein from the New Jersey strain. SEQ ID NO: 13 refers to the ectodomain of the VSV-G protein from the Marraba strain. SEQ ID NO: 15 refers to the ectodomain of the VSV-G protein from the Carajas strain. SEQ ID NO: 17 refers to the ectodomain of the VSV-G protein from the Alagoa strain. Sequence ID 19 refers to the ectodomain of the VSV-G protein from the Cocal strain. Sequence ID 21 refers to the ectodomain of the VSV-G protein from the Morreton strain. Therefore, the residue aligned at residue 182 compared to Sequence ID 2 can also be mutated as provided herein.

[0183] 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.

[0184] In some embodiments, the mutation at position 182 compared to sequence number 2 is I182S, I182H, I182T, I182Q, or I182N. In some embodiments, the mutation at position 182 compared to sequence number 11 is T182S, T182H, T182Q, or T182N. In some embodiments, the mutation at position 182 compared to sequence number 13 is A182S, A182H, A182T, A182Q, or A182N. In some embodiments, the mutation at position 182 compared to sequence number 15 is V182S, V182H, V182T, V182Q, or V182N. In some embodiments, the mutation at position 182 compared to sequence number 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.

[0185] These mutations may be described with reference to SEQ ID NO: 1 or SEQ ID NO: 2, which are VSV-G proteins from the Indiana strain, but these mutations may also be used in other strains of the VSV-G protein. For example, the mutations can be produced in the VSV-G New Jersey strain, the VSV-G Marraba strain, the VSV-G Carajas strain, the VSV-G Alagoa strain, the VSV-G Cocal strain, or the VSV-G Morreton strain. In some embodiments, the respective sequences are as provided herein. Examples of these 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 external domain of the VSV-G New Jersey strain is sequence number 10 and sequence number 11, respectively; the wild-type full-length or external domain of the VSV-G Marraba strain is sequence number 12 and sequence number 13, respectively; the wild-type full-length or external domain of the VSV-G Carajas strain is sequence number 14 and sequence number 15, respectively; the wild-type full-length or external domain of the VSV-G Alagoa strain is sequence number 16 and sequence number 17, respectively; the wild-type full-length or external domain of the VSV-G Cocal strain is sequence number 18 and sequence number 19, respectively; or the wild-type full-length or external domain of the VSV-G Morreton strain is sequence number 20 and sequence number 21, respectively.

[0186] VSV-G proteins containing a 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 (which is incorporated herein by reference in its entirety). For example, VSV-G proteins may contain mutations at positions corresponding to positions 8, 47, 209 and / or 354 of SEQ ID NO: 2.

[0187] In some embodiments, the substitution at position 8 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, excluding Y. In some embodiments, the substitution at position 209 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, excluding H. In some embodiments, the substitution at position 47 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, excluding K or R. In some embodiments, the substitution at position 354 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, excluding K or R.

[0188] In some embodiments, the substitution is at position 47 or 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.

[0189] In some embodiments, the substitution at position 8 is alanine, i.e., H8A.

[0190] In some embodiments, the substitution at position 47 is Q or N, i.e., K47Q or K47N.

[0191] In some embodiments, the protein contains a mutation (substitution) at position 10. In some embodiments, the substitution / mutation is Q10A, Q10R, or Q10K.

[0192] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 2 is 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%, 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 the I182D or I182E mutation. In some embodiments, the VSV-G protein contains the I182S, I182H, I182T, I182Q, or I182N mutation.

[0193] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 11 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) when containing a mutation at position 182. In some embodiments, the polypeptide contains the T182D or T182E mutation. In some embodiments, the VSV-G protein contains the T182S, T182H, T182Q, or T182N mutation.

[0194] 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 compared to SEQ ID NO: 13 (or SEQ ID NO: 12 if a full-length protein is used). In some embodiments, the polypeptide contains the A182D or A182E mutation. In some embodiments, the VSV-G protein contains the A182S, A182H, A182T, A182Q, or A182N mutation.

[0195] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 15 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 using a full-length protein) when containing a mutation at position 182. In some embodiments, the polypeptide contains the V182D or V182E mutation. In some embodiments, the VSV-G protein contains the V182S, V182H, V182T, V182Q, or V182N mutation.

[0196] 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 the V182D or V182E mutation. In some embodiments, the VSV-G protein contains the V182S, V182H, V182T, V182Q, or V182N mutation.

[0197] 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 the V182D or V182E mutation. In some embodiments, the VSV-G protein contains the V182S, V182H, V182T, V182Q, or V182N mutation.

[0198] In some embodiments, a protein containing a mutation at position 182 compared to SEQ ID NO: 21 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) when containing a mutation at position 182. In some embodiments, the polypeptide contains the I182D or I182E mutation. In some embodiments, the VSV-G protein contains the I182S, I182H, I182T, I182Q, or I182N mutation. Viral glycoproteins

[0199] The mutant VSV-G protein can be used, for example, to pseudotype a virus (e.g., a lentivirus). Thus, in some embodiments, viral particles comprising the mutant VSV-G protein provided herein are provided. In some embodiments, the viral particles comprise a VSV-G protein having a mutation at position 198 compared to SEQ ID NO: 1. In some embodiments, a protein having a mutation at position 182 compared to SEQ ID NO: 2 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) with the mutation at position 182. In some embodiments, the polypeptide contains the I182D or I182E mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein contains the I182S, I182H, I182T, I182Q, or I182N mutation.

[0200] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 198 compared to SEQ ID NO: 10. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 11 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 compared to SEQ ID NO: 11. In some embodiments, the VSV-G protein contains T182S, T182H, T182Q, or T182N mutations.

[0201] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 198 compared to SEQ ID NO: 12. In some embodiments, the protein with 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 compared to SEQ ID NO: 13. In some embodiments, the VSV-G protein contains A182S, A182H, A182T, A182Q, or A182N mutations.

[0202] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 203 compared to SEQ ID NO: 14. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 15 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 compared to SEQ ID NO: 15. In some embodiments, the VSV-G protein includes the V182S, V182H, V182T, V182Q, or V182N mutation.

[0203] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 199 compared to SEQ ID NO: 16. In some embodiments, the protein with 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 compared to SEQ ID NO: 17. In some embodiments, the VSV-G protein includes the V182S, V182H, V182T, V182Q, or V182N mutation.

[0204] In some embodiments, the viral particles contain a VSV-G protein with a mutation at position 199 compared to SEQ ID NO: 18. In some embodiments, the protein with 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 compared to SEQ ID NO: 19. In some embodiments, the VSV-G protein includes the V182S, V182H, V182T, V182Q, or V182N mutation.

[0205] In some embodiments, the viral particle contains a VSV-G protein with a mutation at position 199 compared to SEQ ID NO: 20. In some embodiments, the protein with a mutation at position 182 compared to SEQ ID NO: 21 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 the I182D or I182E mutation compared to SEQ ID NO: 21. In some embodiments, the VSV-G protein contains I182S, I182H, I182T, I182Q, or I182N mutations.

[0206] In some embodiments, the VSV-G protein further includes mutations at positions corresponding to positions 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: 1 is T352. In some embodiments, the VSV-G protein includes a mutation corresponding to the T214N mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein includes a mutation corresponding to the T352A mutation compared to SEQ ID NO: 2. In some embodiments, the VSV-G protein includes the T214N and T352A mutations compared to SEQ ID NO: 2. These mutations can be combined with any other mutations 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 NOs. 22 and 23, which combine I182D or I182E with the T214N and T352A mutations, respectively. The sequences are also illustrated below along with the leader sequences that are removed during protein processing. VSV-G protein_I196D, T230N, and T368A mutations (with leader sequence and adjusted numbering) MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQS CGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKD LFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 24) VSV-G protein_I182D, T214N, and T352A mutations (no leader sequence) KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAV IVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLDSMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFP ECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 22) VSV-G protein with I196E, T230N, and T368A mutations (with leader sequence and adjusted numbering) MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQS CGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKD LFAAARFPECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 25) VSV-G protein with I182E, T214N, and T352A mutations (no leader sequence) KFTIVFPHNQKGNWKNVPSNYHYCPSSSDLNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPSVEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAV IVQVTPHHVLVDEYTGEWVDSQFINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLESMDITFFSEDGELSSLGKEGTGFRSNYFAYENGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFP ECPEGSSISAPSQTSVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLKYFETRYIRVDIAAPILSRMVGMISGTTAERELWDDWAPYEDVEIGPNGVLRTSSGYKFPLYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWFSSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK(Sequence ID 23)

[0207] Other strains of the VSV-G protein described herein may also contain mutations corresponding to T214N and / or T352A in SEQ ID NO: 2, as shown in SEQ ID NOs: 22 and 23.

[0208] In some embodiments, the composition includes mutations described in Hwang et al., Gene Ther 2013 Aug; Bren nan et al.; 20(8):807-15. (Epub 2013 Jan 31), which are incorporated herein by reference in their entirety. For example, the mutations may be at positions 230, 368, 66, and / or 162, corresponding to SEQ ID NO: 1. If the leader sequence is removed, the positions are 16 positions fewer compared to SEQ ID NO: 2. In some embodiments, the mutations at those positions are, for example, T230N, T368A, K66T, S162T, or any combination thereof. In some embodiments, the VSV-G protein includes the T230N and T368A mutations. In some embodiments, the VSV-G polypeptide includes K66T, S162T, T230N, and T368A. These positions correspond to the positions in the full-length protein (SEQ ID NO: 1). In some embodiments, the VSV-G protein includes the T230N mutation, the T368A mutation, the K66T mutation, the S162T mutation, or any combination thereof. In some embodiments, the VSV-G protein further includes one or more mutations (e.g., the mutations described in U.S. Patent Application Publication No. 20200216502, which is incorporated herein by reference in its entirety) in addition to the mutation corresponding to position 182 of SEQ ID NO: 2. For example, the VSV-G protein may further include mutations at positions 8, 47, 209 and / or 354 of SEQ ID NO: 2.

[0209] In some embodiments, the substitution at position 8 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, except Y. In some embodiments, the substitution at position 209 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, except H. In some embodiments, the substitution at position 47 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, except K or R. In some embodiments, the substitution at position 354 is by any amino acid other than the amino acid shown at that position in the sequence of SEQ ID NO: 2, except K or R. In some embodiments, the substitution is at position 47 or 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.

[0210] Furthermore, in some embodiments, the virus may be pseudotyped with other viral structural proteins instead of the VSV-G protein or its variants.

[0211] For example, a viral particle can be pseudotyped with the carp spring viremia virus G (SVCV-G) protein and transduction into a cell if the virus contains a targeting moiety. The provided carp spring viremia virus G protein can be used in some embodiments to transduce target cells and deliver heterologous molecules to target cells. In some embodiments, a carp spring viremia virus G protein comprising SEQ ID NO: 52 is provided. SEQ ID NO: 52 is a full-length protein, and SEQ ID NO: 53 is the ectodomain of the carp spring viremia virus G protein with the N-terminal signal peptide removed. Thus, in some embodiments, the protein comprises the amino acid sequence of SEQ ID NO: 53. The carp spring viremia virus G protein can be used to pseudotype viruses such as, for example, lentiviruses, but is not limited to these. Thus, in some embodiments, viral particles comprising the carp spring viremia virus G protein provided herein are provided. In some embodiments, the virus particles contain a carp spring viremia virus G protein comprising a sequence that is 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% identical to the sequence of SEQ ID NO: 52 or SEQ ID NO: 53.

[0212] Sequence of the carp spring viremia virus G (SEQ ID NO: 52 - with leader sequence) MSIISYIAFLLLIDSNLGIPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDDLGF PPQSCGWASVTTVSNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWV EKTAGTLTTIHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSLNGTDPRTNASRVFWDDWELD GNIYQGFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENGNPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSKRPATESHEMRSLV

[0213] Sequence of carp spring viremia virus G (SEQ ID NO: 53 - no leader sequence): IPIFVPSGRNISWQPVIQPFDYQCPIHGNLPNTMGLSATKLTIKSPSVFSTDKVSGWICHAAEWKTTCDYRWYGPQYITHSIHPISPTIDECRRIIQRIASGTDEDLGFPPQSCGWASVTTVSNTNYRVVPHSVHLEPYGGHWIDHEFNGGECREKVCEMKGNHSIWITEETVQHECAKHIEEVEGIMYGNVPRGDVMYANNFIIDRHHRVYRFGGSCQMKFCNKDGIKFARGDWVEKTAGTLTT IHDNVPKCVDGTLVSGHRPGLDLIDTVFNLENVVEYTLCEGTKRKINKQEKLTSVDLSYLAPRIGGFGSVFRVRNGTLERGSTTYIRIEVEGPIVDSNLGTDPRTNASRVFWDDWELDGNIYQGFNGVYKGKDGKIHIPLNMIESGIIDDELQHAFQADIIPHPHYDDDEIREDDIFFDNTGENPVDAVVEWVSGWGTSLKFFGMTLVALILIFLLIRCCVACTYLMKRSPRATESHEMRSLV The target part

[0214] In some embodiments, the viral particle comprises a targeting moiety having the formula T-S1, where T is a target-binding domain and S1 is a stalk moiety. The targeting moiety can be used to target viral particles containing a mutant VSV-G protein or SVCV-G protein to cells expressing a target to which the targeting moiety binds. In some embodiments, the target-binding domain is an antibody, scFv antibody, antigen-binding domain, ankyrin repeat (e.g., DARPIN), VHH domain antibody, nanobody, single-domain antibody, FN3 domain, or any combination thereof. The target-binding domain may be bound to the viral surface via a modified Fc protein provided herein (e.g., L1-Fc-L2-X1) or via a flexible polypeptide provided herein (e.g., L3-X1). In some embodiments, the targeted moiety is bound (fused or ligated) to envelope glycoprotein G or H of a Paramyxoviridae virus, such as a morbillivirus like measles virus, or a henipavirus like nipah virus, cedar virus, or hendra virus. In some embodiments, the targeted moiety may be bound (fused or ligated) to glycoproteins of Rhabdoviridae viruses such as bullous stomatitis New Jersey virus, bullous stomatitis Indiana virus, bullous stomatitis Alagoas virus, bullous interstitial flu Maraba virus, bullous stomatitis Carajas virus, parainfluenza virus, Spodoptera frugiperda rhabdovirus isolate Sf-G, Drosophila obscura Sigmavirus 10A, Wuhan insect virus 7, perch virus, or koi spring viremia virus. In some embodiments, the VSV protein is a mutant protein such as those provided herein. In some embodiments, the targeted portion is bound to a glycoprotein of a filoviridae virus such as Ebola virus, or to a glycoprotein of an arenaviridae virus such as Machupovirus.

[0215] In some embodiments, the target cell is scFv. In some embodiments, the target-binding domain is a single-domain antibody. In some embodiments, the target cell is VHH.

[0216] In some embodiments, the targeted moieties include CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR alpha, TCR beta, TCR gamma, TCR delta, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3; a glycosylated CD43 epitope expressed in acute leukemia or lymphoma but not in hematopoietic progenitor cells; a glycosylated CD43 epitope expressed on non-hematopoietic cancers; A kinase anchor protein 4 (AKAP-4); adrenaline receptor β3 (ADRB3); AFP; and anaplastic lymphoma kinase. kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie2); autoantibody against desmoglein 1 (Dsg 1); autoantibody against desmoglein 3 (Dsg 3); B7H3 (CD276); biotin; bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; cancer / testicular antigen 1 (NY-ESO-1); cancer / testicular antigen 2 (LAGE-LA); carbonic anhydrase IX (CA1X); carcinoembryonic antigen (CEA); CC CTC binding factor (Zinc Finger Protein)-like (BORIS or Brother of the Regulator of lmprinted Sites);CCR4;CD5;CD19;CD20;CD22;CD24;CD30;CD32(FCGR2A);CD33;CD34;CD38;CD44v6;CD72;CD79a;CD79b;CD97;CD99;CD123;CD171;CD179a;CD179b-IGLll;CD200R;CD276 / B7H3;CD300 molecule-like family member f (CD300LF);CDH1-CD324;CDH6;CDH17;CDH19;X chromosome open reading frame 61 (CXORF61); Claudin 6 (CLDN6); Claudin 18.2 (CLD18A2 or CLDN18A.2); CMV pp65; C-MYC epitope tag; Cripto; CS1 (also called 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-containing mucin-like hormone receptor-like molecule 2 (EMR2); Elongation factor 2 variant Muted (ELF2M); Ephrin B2; Ephrin 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-like tyrosine kinase 3 (FLT3); Folate receptor alpha (FRa or FR1); Folate receptor β (FRb); Follicle-stimulating hormone receptor (FSHR); Fos-related antigen 1; Fucosyl-GM1; G protein-coupled receptor class C group 5 member D (GPRC5D); G protein-coupled receptor 20 20, GPR20); GAD; Ganglioside G2 (GD2); Ganglioside GD3 (αNeu 5 Ac(2-8)αNeu 5 Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer); Ganglioside GM3 (αNeu 5 Ac(2-3)bDClalp(1-4)bDGlcp(1-1)Cer); GD3; GFRalpha4; Glycoprotein 100 (gplOO);Glypican-3 (GPC3); Gonadotropin hormone receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylyl cyclase C (GCC); Heat shock protein 70-2 mutated (mut hsp70-2); Hepatitis A virus cellular receptor 1 (HAVCR1); Hexasaccharide portion of globoH glycoceramide (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 papilloma virus E6 (HPV) E6); Human papilloma virus 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 alpha (IL-11Ra); Interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2); Enteral carboxylesterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; Regmine; Leukocyte immunoglobulin-like receptor subfamily A member 2 2. LILRA2); Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); Luteinizing hormone receptor (LHR); Lewis (Y) antigen; Lews Ag; Livl; Gene locus K9 (LY6K); Low conductance chloride channel; Lymphocyte antigen 6 complex;Lymphocyte antigen 75 (LY75); lymphocyte-specific protein tyrosine kinase (LCK); mammary gland differentiation antigen (NY-BR-1); melanoma antigen recognized by T cell 1 (Melanoa or MART 1); melanoma-associated antigen 1 (MAGE-A1); melanoma cancer testis antigen-1 (MAD-CT-1); melanoma cancer testis antigen-2 (MAD-CT-2); melanoma inhibitor of apoptosis (ML-IAP); mesothelin; MPL; mucin 1 cell surface-bound (MUC1); N-acetylglucosaminyl transferase V (NA17); nectin-4; neural cell adhesion molecule (NCAM); NKG2D; NYBR1; O-acetyl-GD2 ganglioside (O-acetyl-GD2 ganglioside (OAcGD2); olfactory receptor 51E2 (OR51E2); oncogene fusion protein consisting of a cleavage cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Ab1) (bcr-ab1); P53 variant; paired box protein Pax-3 (PAX3); paired box protein Pax-5 (PAX5); pannexin 3 (PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor beta (PDGFR-beta); polysialic acid; proacrosin-binding protein sp32 (OY-TES1); prostase; prostate cancer tumor antigen-1 (PCT A-1 or galectin 8); prostate stem cell antigen (PSCA); prostate-specific membrane antigen (PSMA);Prostatic acid phosphatase (PAP); prostain; protease serine 21 (testisin or PRSS21); proteasome (prosome, macropain) subunit beta 9 (LMP2); PTK7; Ras G12V; Ras homolog family member C (RhoC); rat sarcoma (Ras) mutant; progressive glycation end product receptor (RAGE-1); receptor tyrosine kinase-like orphan receptor 1 (ROR1); receptor tyrosine-protein kinase ERBB2 or Her-22 / neu; renal ubiquitous 1 (RU1); renal ubiquitous 2 (RU2); sarcoma translocation breakpoint; serine 2 (TMPRSS2) ETS fusion gene; sialyl Lewis adhesion molecule molecule, sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialyl Lewis antigen); sperm protein 17 (SPA17); squamous cell carcinoma antigen 3 recognized by T cells (SART3); stage-specific embryonic antigen-4 (SSEA-4); STEAP1; Survivin; synovial sarcoma, X breakpoint 2 (SSX2); TCR gamma alternative reading frame protein (TARP); TCR-beta 1 chain; TCR-beta 2 chain; TCR-delta chain; TCR-gamma chain; TCR-gamma-delta; telomerase; TGFbetaR2; 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;It binds to tumor endothelial marker 1 (TEM1 / CD248); tumor endothelial marker 7-related (TEM7R); tumor protein p53 (p53); tumor-associated glycoprotein 72 (TAG72); tyrosinase; tyrosinase-related protein 2 (TRP-2); uroplakin 2 (UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein (WT1); or the X antigen family, member 1A (XAGE1). In some embodiments, the targeting moiety binds to CD7. In some embodiments, the targeting moiety binds to CD8.

[0217] In some embodiments, the targeting portion binds to a target present on a cell, such as an immune cell. In some embodiments, the cell is an immune cell, such as a T cell, B cell, NK cell, dendritic cell, neutrophil, macrophage, cancer cell; or, for example, a CD3+ T cell; CD4+ T cell; CD7+ T cell, CD8+ T cell; CD19+ B cell; CD19+ cancer cell; CD20+ B cell; CD20+ cancer cell; CD30+ lung epithelial cell; CD34+ hematopoietic stem cell; CD105+ endothelial cell; CD105+ hematopoietic stem cell; CD117+ hematopoietic stem cell; CD133+ cancer cell; EpCAM+ cancer cell; GluA2+ neuron; GluA4+ neuron; hematopoietic stem cell; hepatocyte; Her2 / Neu+ cancer cell; NKG2D+ natural killer cell; SLC1A3+ astrocyte; SLC7A10+ adipocyte, etc. In some embodiments, the cell is a T cell. In some embodiments, the cell is a B cell. In some embodiments, the cells are CD7+ T cells and / or CD8+ T cells. CD7-conjugated polypeptide

[0218] In some embodiments, the targeting moiety (e.g., polypeptide) binds to CD7.

[0219] In some embodiments, the polypeptide that binds to CD7 is an antibody that binds to non-human primate CD7. In some embodiments, the polypeptide that binds to CD7 is an antibody that binds to human CD7. The sequence of human CD7 (UniProtKB P09564) is as follows (SEQ ID NO: 29). MAGPPRLLLLPLLLALARGLPGALAAQEVQQSPHCTTVPVGASVNITCSTSGGLRGIYLRQLGPQPQDIIYYEDGVVPTTDRRFRGRIDFSGSQDNLTITMHRLQLSDTGTYTCQAITEVNVYGSGTLVLVTEEQSQGWHRCSDAPPRASALPAPPTGSALPDPQTASALPDPPAASALPAALAVISFLLGLGLGVACVLARTQIKKLCSWRDKNSAACVVYEDMSHSRCNTLSSPNQYQ(Sequence ID 29).

[0220] In some embodiments, the CD7 to which the polypeptide binds is expressed on the surface of the cell. 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, lymphoid progenitor cell, hematopoietic stem cell, myeloid 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-stem cell CD8+CCR7+.

[0221] In some embodiments, the antibody includes an Fc region. The Fc region may be linked to the heavy or light chain of the antibody. In some embodiments, the Fc region is IgG Fc. In some embodiments, IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, IgG Fc is IgG1 Fc. In some embodiments, the antibody includes a constant Fc region as described herein, such as SEQ ID NOs. 26, 27, or 28, or a variant thereof provided herein.

[0222] In some embodiments, polypeptides (e.g., CD7-binding polypeptides) are provided herein. In some embodiments, antibodies (e.g., anti-CD7 antibodies) are provided herein. In some embodiments, the antibody is a recombinant antibody that binds to CD7. In some embodiments, the CD7 protein is a human CD7 protein. In some embodiments, the CD7 protein is a non-human CD7 protein (e.g., mouse, rat, pig, dog, or non-human primate). As used herein, the term “recombinant antibody” refers to an antibody that does not exist in nature. In some embodiments, the term “recombinant antibody” refers to an antibody that has not been isolated from a human subject.

[0223] In some embodiments, an antibody or an antigen-binding fragment thereof is provided, and the antibody or antibody fragment comprises a peptide selected from the following table, which indicates the CDR based on Kabat numbering. TIFF2026521535000002.tif24170

[0224] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to non-human primate CD7. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to human CD7. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having a sequence selected from SEQ ID NOs. 33 to 35. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 33. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 34. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 35. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having a sequence selected from SEQ ID NOs. 30 to 32. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 30. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 31. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 32. CDRs referenced in embodiments throughout this specification can be replaced with CDRs characterized by different forms such as Chothia and IMGT.

[0225] In some embodiments, the polypeptide, antibody, or antibody-conjugated 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.

[0226] In some embodiments, the polypeptide, antibody, or antibody-conjugated 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.

[0227] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises (i) a light chain having any one of the aforementioned enumerated combinations of LCDR1, LCDR2, and LCDR3 sequences; and (ii) a heavy chain having any one of the aforementioned enumerated combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0228] Different CDR motifs can be combined in any combination, including those not shown in the table above. For example, the following embodiments are provided as non-limiting examples of such combinations.

[0229] In some embodiments, a polypeptide, antibody, or antigen-binding fragment thereof includes (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; the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 32; or any of the above-described variants.

[0230] The previous paragraph may refer to CDR under the Kabat system, but equivalent CDR sequences from the naming of IMGT and CHOTHIA can be used.

[0231] 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.

[0232] In some embodiments, the polypeptide comprises a heavy chain variable region peptide or a variant thereof having one of the following sequences: TIFF2026521535000003.tif19170

[0233] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences, or a variant thereof: TIFF2026521535000004.tif19170

[0234] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V of SEQ ID NO: 36 H Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V of SEQ ID NO: 37 L Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V H Peptides and V L Contains peptides, V H The peptide includes the sequence of SEQ ID NO: 36 or a variant thereof; 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 is V H Peptides and V L Contains peptides, V HThe peptide includes the sequence of SEQ ID NO: 36 or a variant thereof; 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 is V H Peptides and V L Contains peptides, V H The peptide includes the sequence of SEQ ID NO: 36 or a variant thereof; 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; V L The peptide contains the sequence of SEQ ID NO: 37.

[0235] V H and V L The array is V H and V L This includes, but is not limited to, the scFv format in which regions are linked by peptide linkers, and may be any other format. 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) is an example, where each n is independently 1 to 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 includes SEQ ID NO: 36 and SEQ ID NO: 37.

[0236] As provided herein, polypeptides, antibodies, or antigen-binding fragments thereof may be sequence variants.

[0237] The polypeptide or antibody sequence may be modified to obtain human IgG antibodies. The sequence transformations provided herein may be modified to obtain other types of antibodies. CDRs can also be linked to other antibodies, proteins, or molecules to create antibody fragments that bind to CD7.

[0238] In some embodiments, the polypeptide or antibody provided herein is a targeting portion on the surface of an engineered viral particle. In some embodiments, the targeting portion enables binding to target cells. In some embodiments, the targeting portion is a CD7 binding portion, such as the polypeptide or antibody provided herein. In some embodiments, the target-binding domain ("T") includes 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: 38: DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS(Sequence ID 38) Alternatively, it may be substantially similar to or an active fragment of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") includes the sequence described in SEQ ID NO: 38. In some embodiments, the target-binding domain ("T") described 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.

[0239] In some embodiments, the polypeptide or antibody 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 enables binding to a target cell. In some embodiments, the targeting portion is a CD7 binding portion, such as a polypeptide or antibody provided herein. In some embodiments, the target-binding domain ("T") includes 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: 39. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 39. QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (Sequence ID 39) Alternatively, it may be substantially similar to or an active fragment of SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") includes the sequence described in SEQ ID NO: 39. In some embodiments, the target-binding domain ("T") described 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. CD8-bound polypeptide

[0240] In some embodiments, the targeting moiety (e.g., polypeptide) can be bound to CD8.

[0241] In some embodiments, the polypeptide binds to CD8. In some embodiments, the polypeptide binds to CD8-α. In some embodiments, the polypeptide binds to CD8-beta. In some embodiments, the polypeptide binds to a CD8 heterodimer. In some embodiments, the CD8 heterodimer contains CD8-alpha and CD8-beta subunits. In some embodiments, the polypeptide binds to a CD8-alpha homodimer. In some embodiments, the polypeptide that binds to CD8 is an antibody that binds to non-human primate CD8. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to non-human primate CD8-α. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to non-human primate CD8-beta. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to a non-human primate CD8-α homodimer. In some embodiments, the antibody that binds to non-human primate CD8 is an antibody that binds to a non-human primate CD8 heterodimer. In some embodiments, the polypeptide that binds to CD8 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-alpha. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-beta. In some embodiments, the antibody that binds to human CD8 is an antibody that binds to human CD8-alpha 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-alpha (UniProtKB Q8TAW8) is as follows (SEQ ID NO: 40): MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGCYFCSALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV (Sequence ID 40).

[0242] The sequence of human CD8-beta (UniProtKB Q8TD28) is as follows (sequence number 41): MRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKISLSNMRIYWLRQRQAPSSDSHHEFLALWDSAKGTIHGEEVEQEKIAVFRDASRFILNLTSVKPEDSGIYFCMIVGSPELTFGKGTQLSVVDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCSPITLGLLVAGVLVLLVSLGVAIHLCCRRRRARLRFMKQLYK (Sequence ID 41).

[0243] In some embodiments, the CD8 to which the polypeptide binds is expressed on the surface of the cell. 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, alpha-beta T cell, gamma-delta T cell, lymphoid progenitor 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.

[0244] In some embodiments, the antibody includes an Fc region. The Fc region may be linked to the heavy or light chain of the antibody. In some embodiments, the Fc region is IgG Fc. In some embodiments, IgG is selected from IgG1, IgG2, IgG3, or IgG4. In some embodiments, IgG Fc is IgG1 Fc. In some embodiments, the antibody includes a constant Fc region as described herein, such as SEQ ID NOs: 26, 27, or 28 or a variant thereof.

[0245] In some embodiments, polypeptides (e.g., CD8-conjugated polypeptides) are provided herein. In some embodiments, antibodies (e.g., anti-CD8 antibodies) are provided herein. 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 does not exist in nature. In some embodiments, the term “recombinant antibody” refers to an antibody that has not been isolated from a human subject.

[0246] In some embodiments, an antibody or an antigen-binding fragment thereof is provided, and the antibody or antibody fragment comprises a peptide selected from the following table, which indicates the CDR based on Kabat numbering. TIFF2026521535000005.tif24170

[0247] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to non-human primate CD8. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain or light chain CDR provided in the table above and binds to human CD8. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having a sequence selected from SEQ ID NOs. 45 to 47. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 45. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 46. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a light chain CDR having the sequence of SEQ ID NO. 47. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having a sequence selected from SEQ ID NOs. 42 to 44. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 42. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 43. In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain CDR having the sequence of SEQ ID NO: 44. The CDRs referenced in embodiments throughout this specification can be replaced with CDRs characterized by different forms such as Chothia and IMGT.

[0248] In some embodiments, the polypeptide, antibody, or antibody-conjugated 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.

[0249] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises a heavy chain variable region having HCDR1, HCDR2, and HCDR3, where 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.

[0250] In some embodiments, the polypeptide, antibody, or antibody-conjugated fragment comprises (i) a light chain having any one of the aforementioned enumerated combinations of LCDR1, LCDR2, and LCDR3 sequences; and (ii) a heavy chain having any one of the aforementioned enumerated combinations of HCDR1, HCDR2, and HCDR3 sequences.

[0251] Different CDR motifs can be combined in any combination, including those not shown in the table above. For example, the following embodiments are provided as non-limiting examples of such combinations.

[0252] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof includes (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; the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 44; or any of the above-described modifications.

[0253] The previous paragraph may refer to CDR under the Kabat system, but equivalent CDR sequences from the naming of IMGT and CHOTHIA can be used.

[0254] 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.

[0255] In some embodiments, the polypeptide comprises a heavy chain variable region peptide or a variant thereof having one of the following sequences: TIFF2026521535000006.tif23170

[0256] In some embodiments, the polypeptide comprises a light chain variable region peptide having one of the following sequences, or a variant thereof: TIFF2026521535000007.tif19170

[0257] In some embodiments, the polypeptide, antibody, or antigen-binding fragment thereof is V of SEQ ID NO: 48 H Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V of SEQ ID NO: 49 L Contains peptides. In some embodiments, polypeptides, antibodies, or antigen-binding fragments thereof are V H Peptides and V L Contains peptides, V H The peptide comprises the sequence of SEQ ID NO: 48 or a variant thereof; 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 is V H Peptides and V L Contains peptides, V HThe peptide comprises the sequence of SEQ ID NO: 48 or a variant thereof; 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 is V H Peptides and V L Contains peptides, V H The peptide comprises the sequence of SEQ ID NO: 48 or a variant thereof; 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; V L The peptide contains the sequence of SEQ ID NO: 49.

[0258] V H and V L The array is V H and V L The peptide linker may be any form, including but not limited to the scFv form in which the region is linked by 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 to 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 includes SEQ ID NOs: 48 and 49.

[0259] As provided herein, polypeptides, antibodies, or antigen-binding fragments thereof may be sequence variants.

[0260] The polypeptide or antibody sequence may be modified to obtain human IgG antibodies. The sequence transformations provided herein may be modified to obtain other types of antibodies. CDRs can also be linked to other antibodies, proteins, or molecules to create antibody fragments that bind to CD8.

[0261] In some embodiments, the polypeptide or antibody provided herein is a targeting portion on the surface of an engineered viral particle. In some embodiments, the targeting portion enables binding to target cells. In some embodiments, the target-binding domain ("T") is a CD8-binding portion, such as the polypeptide or antibody provided herein. In some embodiments, the target-binding domain ("T") includes 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: 50 NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS(Sequence ID 50) Alternatively, it is substantially similar to SEQ ID NO: 50, or an active fragment of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") described in SEQ ID NO: 50 includes the sequence described in SEQ ID NO: 50. In some embodiments, the target-binding domain ("T") described 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.

[0262] In some embodiments, the polypeptide or antibody provided herein is a targeting moiety 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 moiety enables binding to target cells. In some embodiments, the target-binding domain ("T") is a CD8-binding moiety, such as the polypeptide or antibody provided herein. The target-binding domain ("T") includes 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: 51: EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR (Sequence ID 51) Alternatively, it is substantially similar to SEQ ID NO: 51, or is an active fragment of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 90% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 95% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes a sequence that is at least 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") includes the sequence described in SEQ ID NO: 51. In some embodiments, the target-binding domain ("T") described 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. Targeted portion including the Fc domain

[0263] In some embodiments, V H and V LThe polypeptide is linked to a stalk portion S1 containing an Fc region. In some embodiments, the Fc region is as provided herein. In some embodiments, the Fc region is a modified Fc region provided herein. Non-exclusive mutations in the Fc region are provided herein. In some embodiments, the modified Fc region includes a sequence that is a variant of SEQ ID NO: 26, SEQ ID NO: 27, or SEQ ID NO: 28 provided herein. In some embodiments, the variant of SEQ ID NO: 26 includes one or more mutations selected from the group consisting of L234A, L235A, N297A, P329G, I253A, H310A, and H435A provided herein. In some embodiments, the variant of SEQ ID NO: 27 includes one or more mutations selected from the group consisting of N297A, P329G, I253A, H310A, and H435A 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 provided herein. As provided herein, the heavy chain may be linked to the Fc region. In some embodiments, the Fc region further comprises (e.g., linked to) a transmembrane domain. In some embodiments, the Fc region further comprising the transmembrane domain has the formula L1-Fc-L2-X1, where L1 is or is absent a linker provided herein, Fc is the variant Fc region provided herein, L2 is or is absent a linker provided herein, and X1 is a polypeptide comprising the transmembrane domain provided herein. As provided herein, X1 is of the formula ECD-T M -May contain a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M The transmembrane domain provided herein is the transmembrane domain, and the ICD is either an intracellular domain provided herein or absent. Examples of ECDs include, but are not limited to, the CD8 and / or CD28 extracellular domains provided herein. MExamples include, but are not limited to, the CD8 and / or CD28 transmembrane domains provided herein. In some embodiments, X1 is T M It includes, and ECD and ICD are absent. In some embodiments, X1 is ECD, T M It includes, and ICD is not present. In some embodiments, X1 is T M and includes ICD, but does not include ECD. In some embodiments, X1 is ECD, T M , and ICD. It should be understood that in any of the following embodiments, the ECD, ICD, or both may be optionally omitted. Thus, X1 may be CD8 and / or CD28 ECD, CD8 and / or CD28 T M Embodiments including an ICD with an Env embedded motif are understood to include the following X1 members: i) CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and ICDs including the Env built-in motif; ii) CD8 and / or CD28 T M , and ICDs including Env embedded motifs in which no ECD exists;iii) CD8 and / or CD28 ECDs in which no ICD exists, and CD8 and / or CD28 T M ; and iv) CD8 and / or CD28 T in which neither ECD nor ICD exists M Similarly, X1 is CD8 and / or CD28 T M Embodiments including an ICD with an Env embedded motif are understood to include the following X1 members: i) CD8 and / or CD28 T M ICDs including the Env built-in motif; and ii) CD8 and / or CD28 T for which no ICD exists. M Similarly, X1 is CD8 and / or CD28 ECD and CD8 and / or CD28 T M Embodiments including the following X1 members are understood to include: i) CD8 and / or CD28 ECD and CD8 and / or CD28 T M ii) CD8 and / or CD28 T where ECD does not existM Furthermore, the above explanation applies to specific ECDs, T M It should be understood that this also applies to embodiments in which ICD is not described. For example, if X1 is ECD, CD8 and / or CD28 T M Embodiments including ICD are understood to include the following X1 members: i) ECD, CD8 and / or CD28 T M and ICD;ii) ECDs for which no ICD exists, and CD8 and / or CD28 T M iii) CD8 and / or CD28 T for which ECD does not exist M , ICD; and iv) CD8 and / or CD28 T for which ECD and ICD are not present. M Similarly, X1 is CD8 and / or CD28 T M Furthermore, embodiments including the ICD are understood to include the following X1 members: i) CD8 and / or CD28 T M and ICD; and ii) CD8 and / or CD28 T for which no ICD exists. M Similarly, X1 is ECD and CD8 and / or CD28 T M Embodiments including the following X1 members are understood to include: i) ECD and CD8 and / or CD28 T M ii) ECD does not exist CD8 and / or CD28 T M Unless otherwise explicitly stated, the above embodiments and descriptions are applicable to any of the following embodiments.

[0264] In some embodiments, X1 is CD8 and / or CD28 ECD, T M , and ICD. In some embodiments, X1 is ECD, CD8 and / or CD28 T M , and ICD. In some embodiments, X1 is CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and ICD. In some embodiments, X1 is ECD, T M, and ICD, the ICD includes the env embedded motif provided herein. In some embodiments, X1 is CD8 and / or CD28 ECD, T M , and ICD, the ICD includes the env embedded motif provided herein. In some embodiments, X1 is ECD, CD8 and / or CD28 T M , and ICD, the ICD includes the env embedded motif provided herein. In some embodiments, X1 is CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and ICD, the ICD including the env embedded motif provided herein.

[0265] In some embodiments, V provided herein H and V L The polypeptide is linked to a stalk portion (S1) containing an Fc region provided herein. In some embodiments, V provided herein H and V L The polypeptide is linked to a stalk portion (S1) which includes an Fc region containing a transmembrane domain provided herein. In some embodiments, the Fc region further containing a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a modified Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide containing a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M -May contain a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof, which may or may not be provided herein. M V is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein or not provided herein. In some embodiments, V provided herein H and V L Polypeptides are CD8 and / or CD28 ECD, T M, and are connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD. In some embodiments, V provided herein H and V L Polypeptides are ECD, CD8 and / or CD28 T M , and is connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V provided herein H and V L Polypeptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and is connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V provided herein H and V L Polypeptides are ECD, T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V provided herein H and V L Polypeptides are CD8 and / or CD28 ECD, T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V provided herein H and V L Polypeptides are ECD, CD8 and / or CD28 T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V provided herein H and V L Polypeptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M, and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, the ECD, T M , and the V provided herein, connected to the stalk portion (S1) which includes the Fc region (L1-Fc-L2-X1) including the ICD H and V L Polypeptides are immobilized on 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 embodiments, V H and V L Polypeptides bind to immune cells, such as those provided herein.

[0266] In some embodiments, V has the sequence described in Sequence ID No. 36. H A peptide and V having the sequence described in Sequence ID No. 37 L Peptides are ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 36. H A peptide and V having the sequence described in Sequence ID No. 37 L The peptide is CD8 and / or CD28 ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 36. H A peptide and V having the sequence described in Sequence ID No. 37 L The peptides are ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 36. H A peptide and V having the sequence described in Sequence ID No. 37 LThe peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 36. H A peptide and V having the sequence described in Sequence ID No. 37 L Peptides are ECD, T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 36. H A peptide and V having the sequence described in Sequence ID No. 37 L The peptide is CD8 and / or CD28 ECD, T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 36. H A peptide and V having the sequence described in Sequence ID No. 37 L The peptides are ECD, CD8 and / or CD28 T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 36. H A peptide and V having the sequence described in Sequence ID No. 37 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, the ECD, T M V having the sequence described in Sequence ID No. 36, and linked to the stalk portion (S1) including the Fc region (L1-Fc-L2-X1) containing the ICD. HA peptide and V having the sequence described in Sequence ID No. 37 L The peptides are immobilized on 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 embodiments, V H and V L Polypeptides bind to immune cells, such as those provided herein.

[0267] In some embodiments, V includes 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 sequence number 36. H Peptide; and V containing 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: 37. L The peptide is ligated to a stalk moiety (S1) containing an Fc region provided herein. In some embodiments, a V containing 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 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: 37. L The peptide is linked to a stalk moiety (S1) containing an Fc region containing a transmembrane domain provided herein. In some embodiments, the Fc region further containing a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or Fc is a modified Fc region provided herein, L2 is a linker provided herein or absent, and X1 is a polypeptide containing a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M-May contain a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof, which may or may not be provided herein. M V is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein or not provided herein. In some embodiments, V includes 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 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: 37. L Peptides are ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes 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 sequence number 36. H Peptide; and V containing 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: 37. L The peptide is CD8 and / or CD28 ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including the ICD. In some embodiments, V includes 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 sequence number 36. H Peptide; and V containing 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: 37. L The peptides are ECD, CD8 and / or CD28 T M, and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes 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 sequence number 36. H Peptide; and V containing 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: 37. L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes 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 sequence number 36. H Peptide; and V containing 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: 37. L Peptides are ECD, T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, the ICD including an env embedded motif provided herein. In some embodiments, a V containing 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 sequence number 36. H Peptide; and V containing 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: 37. L The peptide is CD8 and / or CD28 ECD, T M, and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, the ICD including an env embedded motif provided herein. In some embodiments, a V containing 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 sequence number 36. H Peptide; and V containing 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: 37. L The peptides are ECD, CD8 and / or CD28 T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, the ICD including an env embedded motif provided herein. In some embodiments, a V containing 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 sequence number 36. H Peptide; and V containing 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: 37. L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, the ICD including an env embedded motif provided herein. In some embodiments, a V containing 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 sequence number 36. H Peptides; and ECD, T MV includes 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 sequence number 37 linked to the stalk portion (S1) including the Fc region (L1-Fc-L2-X1) containing the ICD. L The peptide is immobilized on 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 embodiments, V H and V L Polypeptides bind to immune cells, such as those provided herein.

[0268] In some embodiments, V H Peptides and V L A polypeptide containing a peptide, V H The peptide contains 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; V L A polypeptide is provided, comprising a peptide whose sequence 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: 37.

[0269] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 37. However, V H Peptides and VL The peptide comprises a light chain CDR having the sequence of SEQ ID NOs. 33-35; and / or a heavy chain CDR having the sequence of SEQ ID NOs. 30-32. In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 37. However, V H Peptides and V 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 CDR in the chain is as described in the combinations provided herein.

[0270] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 37. However, 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. 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.

[0271] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 37. However, V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 33 and containing at most one conservative amino acid substitution, LCDR2 having the sequence of SEQ ID NO: 34 and containing at most one conservative amino acid substitution, and LCDR3 having the sequence of SEQ ID NO: 35 and containing at most one conservative amino acid substitution; V H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 30 and containing at most one conservative amino acid substitution, HCDR2 having the sequence of SEQ ID NO: 31 and containing at most one conservative amino acid substitution, and HCDR3 having the sequence of SEQ ID NO: 32 and containing at most one conservative amino acid substitution.

[0272] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 36, V L The peptide contains the sequence of SEQ ID NO: 37.

[0273] In some embodiments, the polypeptides provided herein bind to non-human primate CD7. In some embodiments, the polypeptides provided herein bind to human CD7.

[0274] As provided herein, different polypeptides (V H or V L ) may or may not be linked to a peptide linker, and may instead be a continuous sequence. In some embodiments, the peptide linker is (GGGGS) n The sequence (SEQ ID NO: 55) is included, where each n is independently 1 to 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 is V H -ZV L or V L -ZV H It can be expressed by the formula, where Z is a peptide linker. In some embodiments, Z is (GGGGS) n (Sequence ID 55) where each n is independently 1 to 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.

[0275] In some embodiments, formula V L -ZV H A polypeptide comprising the linked peptide represented by includes the heavy chain variable region described in SEQ ID NO: 36, linked to the light chain variable region described in SEQ ID NO: 37 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, via the peptide linker V H V connected to L The polypeptide containing has the sequence described below. DILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSS (Sequence ID 38).

[0276] In some embodiments, the polypeptide contains 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: 38. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 38. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 38. In some embodiments, the polypeptide described in SEQ ID NO: 38 is an antibody or its antigen-binding fragment. 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.

[0277] In some embodiments, formula V H -ZV L A polypeptide comprising the linked peptide represented by includes a light chain variable region described in SEQ ID NO: 37 linked to the heavy chain variable region described in SEQ ID NO: 36 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, via the peptide linker V L V connected to HThe polypeptide containing has the sequence described below. QVQLQQPGAELVKPGASVKLSCKASGYPFTSYWIHWVKQRPGRGLEWLGRIDPNSGDTKYNEKFKNKATLTVDKSSTTAYMQLSSLTSEDSAVYYCARSPYYSNDNSMDYWGQGTSVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNDSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQSNSWPTTFGGGTKLEIKR (Sequence ID 39).

[0278] In some embodiments, the polypeptide contains 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: 39. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 39. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 39. In some embodiments, the polypeptide described in SEQ ID NO: 39 is an antibody or its antigen-binding fragment. 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.

[0279] In some embodiments, the polypeptide 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: 38, and comprises a stalk portion (S1) that includes an Fc region, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a modified Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M -May contain a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide 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: 38, and includes ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains 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: 38, and includes CD8 and / or CD28 ECD, T M, and a stalk portion (S1)D including an Fc region (L1-Fc-L2-X1) containing IC. In some embodiments, the polypeptide includes 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: 38, and includes ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains 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: 38, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains 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: 38, and includes ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes 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: 38, and includes CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes 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: 38, and includes an ECD, CD8 and / or CD28 T M, and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes 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: 38, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the sequence includes an ECD, T which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 38. M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, 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 embodiments, the polypeptide binds to immune cells such as those provided herein.

[0280] In some embodiments, the polypeptide 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: 39, and comprises a stalk portion (S1) that includes an Fc region, such as those provided herein. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a modified Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T M -May contain a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof, which may or may not be provided herein. M ECD is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein or not provided herein. In some embodiments, the polypeptide 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: 39, and ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains 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: 39, and includes CD8 and / or CD28 ECD, T M, and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains 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: 39, and includes ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains 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: 39, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide contains 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: 39, and includes ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes 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: 39, and includes CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes 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: 39, and includes an ECD, CD8 and / or CD28 T M, and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes 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: 39, and includes CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the sequence includes an ECD, T which is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of sequence number 39. M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, 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 embodiments, the polypeptide binds to immune cells such as those provided herein.

[0281] In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 38 and includes a stalk portion (S1) including an Fc region, such as those provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 38 and includes a stalk portion (S1) including an Fc region, the Fc region further includes a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further including a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a modified Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide including a transmembrane domain provided herein. As provided herein, X1 is of the formula ECD-T M -May contain a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof provided herein, or is absent. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 38, ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 38, CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 38, ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 38, and CD8 and / or CD28 ECD, CD8 and / or CD28 T M, and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 38, ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 38, CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 38, ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1)) including an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 38, CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the sequence includes the sequence described in Sequence ID No. 38, and the ECD, T M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, 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 embodiments, the polypeptide binds to immune cells such as those provided herein.

[0282] In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 39 and includes a stalk portion (S1) including an Fc region, such as those provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 39 and includes a stalk portion (S1) including an Fc region, the Fc region further includes a transmembrane domain, such as those provided herein. In some embodiments, the Fc region further including a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a modified Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide including a transmembrane domain provided herein. As provided herein, X1 is of the formula ECD-T M -May contain a polypeptide having an ICD, wherein the ECD is an extracellular domain or fragment thereof, which may or may not be provided herein. M is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein or not provided herein. In some embodiments, the polypeptide comprises a sequence having the sequence described in SEQ ID NO: 39, ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 39, CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 39, ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 39, and CD8 and / or CD28 ECD, CD8 and / or CD28 T M, and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including ICD. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 39, ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 39, CD8 and / or CD28 ECD, T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, wherein the ICD includes the env embedded motif provided herein. In some embodiments, the polypeptide includes a sequence having the sequence described in SEQ ID NO: 39, ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the polypeptide includes a sequence having the sequence described in Sequence ID No. 39, CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, wherein the ICD includes an env embedded motif provided herein. In some embodiments, the sequence includes the sequence described in Sequence ID No. 39, and the ECD, T M A polypeptide comprising a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing ICD is immobilized on the surface of a viral particle, 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 embodiments, the polypeptide binds to immune cells such as those provided herein.

[0283] In some embodiments, polypeptides provided herein that include the formula T-S1 include an amino acid sequence in which T is a target-binding domain and S1 is a stalk moiety, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 98. (Sequence ID 98) Alternatively, it may be substantially similar to or an active fragment of SEQ ID NO: 98. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 98. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98. In some embodiments, the polypeptide provided herein, comprising the formula T-S1, comprises the amino acid sequence of SEQ ID NO: 98.

[0284] In some embodiments, polypeptides provided herein that include the formula T-S1 include an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 126, where T is a target-binding domain and S1 is a stalk portion. (Sequence ID 126)

[0285] In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 126. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 126. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 126. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises the amino acid sequence of SEQ ID NO: 126.

[0286] In some embodiments, polypeptides provided herein that include the formula T-S1 include an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 127, where T is a target-binding domain and S1 is a stalk portion. (Sequence ID 127) In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 127. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 127. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 127. In some embodiments, the polypeptide provided herein, comprising formula T-S1, comprises the amino acid sequence of SEQ ID NO: 127.

[0287] In some embodiments, a polypeptide provided herein, comprising the formula T-S1, wherein T is a target-binding domain and S1 is a stalk portion, comprises an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% 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 the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptides provided herein, comprising the formula T-S1, comprise an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, where the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptides provided herein, comprising the formula T-S1, comprise an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, where the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptides provided herein, comprising the formula T-S1, comprise the amino acid sequence of SEQ ID NO: 98, where the target-binding domain "T" of the polypeptide corresponds to amino acids 25-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98.

[0288] In some embodiments, polypeptides provided herein comprising the formula T-S1, wherein T is V H and V LThe target-binding domain, which includes S1, is a polypeptide whose stalk portion contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 98, and the target-binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L The T-S1 portion corresponds to amino acids 172-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptides provided herein that include the formula T-S1 have an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 98, and the V portion of the target-binding domain H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L The T-S1 portion corresponds to amino acids 172-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptides provided herein that include the formula T-S1 have an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, and the V portion of the target-binding domain H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L The stalk portion "S1" of the polypeptide corresponds to amino acids 172-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide provided herein comprising formula T-S1 comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L The T-S1 portion corresponds to amino acids 172-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98. In some embodiments, the polypeptide provided herein, which includes the formula T-S1, comprises the amino acid sequence of SEQ ID NO: 98, and the V portion of the target-binding domain HThis corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L This corresponds to amino acids 172-283 of SEQ ID NO: 98, and the stalk portion "S1" of the polypeptide corresponds to amino acids 284-634 of SEQ ID NO: 98.

[0289] In some embodiments, a polypeptide provided herein is provided, comprising the formula T-L1-Fc-L2-X1, where T is a target-binding domain, L1 is a polypeptide linker or absent, Fc is a modified Fc domain provided herein, L2 is a polypeptide linker or absent, and X1 is a polypeptide comprising a transmembrane domain, comprising an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% 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, 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 polypeptides provided herein comprising the formula T-L1-Fc-L2-X1 comprises 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 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 polypeptides provided herein comprising the formula T-L1-Fc-L2-X1 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, 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 provided herein, comprising the formula T-L1-Fc-L2-X1, 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, 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.

[0290] In some embodiments, the polypeptides provided herein include the formula T-L1-Fc-L2-X1, where T is V H and V L A target-binding domain comprising L1 is a polypeptide linker or absent, Fc is a modified Fc domain provided herein, L2 is a polypeptide linker or absent, X1 is a polypeptide comprising a transmembrane domain and comprising an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 98, and the target-binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. LA polypeptide is provided in which L1 corresponds to amino acids 172-283 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 provided herein comprising the formula T-L1-Fc-L2-X1 comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 98, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 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 polypeptides provided herein comprising the formula T-L1-Fc-L2-X1 comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, and the target-binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 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 polypeptides provided herein comprising the formula T-L1-Fc-L2-X1 comprises an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, and the target-binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 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 provided herein comprising the formula T-L1-Fc-L2-X1 comprises the amino acid sequence of SEQ ID NO: 98, and the target-binding domain VH This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds to amino acids 172-283 of sequence number 98, L1 contains amino acids 284-301 of sequence number 98, Fc contains amino acids 302-542 of sequence number 98, L2 is absent, and X1 corresponds to amino acids 543-634 of sequence number 98.

[0291] In some embodiments, T-L1-Fc-L2-ECD-T M A polypeptide provided herein comprising the formula -ICD, wherein T is a target-binding domain, L1 is a polypeptide linker or is absent, Fc is a modified Fc domain provided herein, L2 is a polypeptide linker or is absent, ECD is an extracellular domain, and T M ICD is a transmembrane domain, ICD is an intracellular domain containing an env embedded motif, and contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 98, the polypeptide target binding domain "T" 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, T M A polypeptide is provided in which the formula T-L1-Fc-L2-ECD-T contains amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the formula T-L1-Fc-L2-ECD-T is provided. M- The polypeptides provided herein, including ICD, contain 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 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 component contains amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, contain 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, 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 formula T-L1-Fc-L2-ECD-T includes amino acids 585-612 of SEQ ID NO: 98, the ICD includes amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif includes amino acids 627-634 of SEQ ID NO: 98. In some embodiments, the formula T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including ICD, contain an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, 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, T M The component contains amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M- The polypeptides provided herein, including the formula ICD, comprise the amino acid sequence of SEQ ID NO: 98, 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, ECD comprises amino acids 543-584 of SEQ ID NO: 98, T M The first element contains amino acids 585-612 of SEQ ID NO: 98, the second element contains amino acids 613-634 of SEQ ID NO: 98, and the third element (env) contains amino acids 627-634 of SEQ ID NO: 98.

[0292] In some embodiments, T-L1-Fc-L2-ECD-T M A polypeptide provided herein comprising the formula -ICD, wherein T is V H and V L A target-binding domain comprising, L1 is a polypeptide linker or absent, Fc is a modified Fc domain provided herein, L2 is a polypeptide linker or absent, ECD is an extracellular domain, T M ICD is a transmembrane domain, and ICD is an intracellular domain containing an env embedded motif, and contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 98, and the V of the target binding domain H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds 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, T MA polypeptide is provided in which the is composed of amino acids 585-612 of SEQ ID NO: 98, the ICD is composed of amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif is composed of amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 98, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds 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, T M The component contains amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, and the target binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds 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, T M The component contains amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, and the target binding domain V HThis corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds 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, T M The component contains amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptide provided herein, comprising the formula ICD, comprises the amino acids of SEQ ID NO: 98, and the target-binding domain V H This corresponds to amino acids 25-150 of sequence number 98, and is the V of the target-binding domain. L L1 corresponds 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, T M The first element contains amino acids 585-612 of SEQ ID NO: 98, the second element contains amino acids 613-634 of SEQ ID NO: 98, and the third element (env) contains amino acids 627-634 of SEQ ID NO: 98.

[0293] In some embodiments, the polypeptides provided herein are of the formula T-L1-Fc-L2-ECD-T M - Containing ICD, where T is a target-binding domain, L1 is a polypeptide linker or is absent, Fc is a modified Fc domain provided herein, L2 is a polypeptide linker or is absent, ECD is an extracellular domain, T ML1 is a transmembrane domain, ICD is an intracellular domain containing an env embedded motif, and contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 98, the polypeptide target binding domain "T" contains the amino acid sequence of 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 of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, comprise an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 98, the target-binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39, corresponding to amino acids 25-283 of SEQ ID NO: 98, L1 comprises the amino acid sequence of SEQ ID NO: 72, corresponding to amino acids 284-301 of SEQ ID NO: 98, Fc comprises the amino acid sequence of SEQ ID NO: 104, corresponding to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60, corresponding to amino acids 543-584 of SEQ ID NO: 98, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M- The polypeptides provided herein, including the formula ICD, comprise an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, the target-binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39, corresponding to amino acids 25-283 of SEQ ID NO: 98, L1 comprises the amino acid sequence of SEQ ID NO: 72, corresponding to amino acids 284-301 of SEQ ID NO: 98, Fc comprises the amino acid sequence of SEQ ID NO: 104, corresponding to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60, corresponding to amino acids 543-584 of SEQ ID NO: 98, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, comprise an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, the target-binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39, corresponding to amino acids 25-283 of SEQ ID NO: 98, L1 comprises the amino acid sequence of SEQ ID NO: 72, corresponding to amino acids 284-301 of SEQ ID NO: 98, Fc comprises the amino acid sequence of SEQ ID NO: 104, corresponding to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60, corresponding to amino acids 543-584 of SEQ ID NO: 98, T M The amino acid sequence of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M- The polypeptide provided herein, including the formula ICD, comprises the amino acid sequence of SEQ ID NO: 98, the target-binding domain "T" of the polypeptide comprises the amino acid sequence of SEQ ID NO: 39, corresponding to amino acids 25-283 of SEQ ID NO: 98, L1 comprises the amino acid sequence of SEQ ID NO: 72, corresponding to amino acids 284-301 of SEQ ID NO: 98, Fc comprises the amino acid sequence of SEQ ID NO: 104, corresponding to amino acids 302-542 of SEQ ID NO: 98, L2 is absent, ECD comprises the amino acid sequence of SEQ ID NO: 60, corresponding to amino acids 543-584 of SEQ ID NO: 98, T M The sequence contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98; the ICD contains amino acids 613-634 of SEQ ID NO: 98; and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98.

[0294] In some embodiments, T-L1-Fc-L2-ECD-T M A polypeptide provided herein comprising the formula -ICD, wherein T is V H and V L A target-binding domain comprising, L1 is a polypeptide linker or absent, Fc is a modified Fc domain provided herein, L2 is a polypeptide linker or absent, ECD is an extracellular domain, T M ICD is a transmembrane domain, and ICD is an intracellular domain containing an env embedded motif, and contains an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 98, and the V of the target binding domain H This contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 98, and the target binding domain V LL1 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 polypeptide comprises the amino acid sequence of SEQ ID NO: 62, corresponding to amino acids 585-612 of SEQ ID NO: 98; the ICD comprises amino acids 613-634 of SEQ ID NO: 98; and the env embedded motif comprises the amino acid sequence of SEQ ID NO: 63, corresponding to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 98, and the target binding domain V H This contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 98, and the target binding domain V L L1 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 of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 98, and the target binding domain V HThis contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 98, and the target binding domain V L L1 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 of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptides provided herein, including the formula ICD, have an amino acid sequence that is at least 98% identical to the sequence of SEQ ID NO: 98, and the target binding domain V H This contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 98, and the target binding domain V L L1 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 of SEQ ID NO: 62 corresponds to amino acids 585-612 of SEQ ID NO: 98, the ICD contains amino acids 613-634 of SEQ ID NO: 98, and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 corresponds to amino acids 627-634 of SEQ ID NO: 98. In some embodiments, T-L1-Fc-L2-ECD-T M - The polypeptide provided herein, which includes the formula ICD, comprises the amino acid sequence of SEQ ID NO: 98, and the target binding domain VH This contains the amino acid sequence of SEQ ID NO: 36, corresponding to amino acids 25-150 of SEQ ID NO: 98, and the target binding domain V L L1 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 sequence contains the amino acid sequence of SEQ ID NO: 62 and corresponds to amino acids 585-612 of SEQ ID NO: 98; the ICD contains amino acids 613-634 of SEQ ID NO: 98; and the env embedded motif contains the amino acid sequence of SEQ ID NO: 63 and corresponds to amino acids 627-634 of SEQ ID NO: 98.

[0295] In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L Peptides are ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptide is CD8 and / or CD28 ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptides are ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V has the sequence described in sequence number 48. H Peptide and V having the sequence described in Sequence ID No. 49 L Peptides are ECD, T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptide is CD8 and / or CD28 ECD, T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptides are ECD, CD8 and / or CD28 T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, V has the sequence described in Sequence ID No. 48. H Peptide and V having the sequence described in Sequence ID No. 49 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and connected to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD including an env embedded motif provided herein. In some embodiments, the ECD, T M V having the sequence described in sequence number 48, and linked to the stalk portion (S1) which includes the Fc region (L1-Fc-L2-X1) including the ICD. HPeptide and V having the sequence described in Sequence ID No. 49 L The peptides are immobilized on 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 embodiments, V H and V L The peptides bind to immune cells, such as those provided herein.

[0296] In some embodiments, V includes 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 sequence number 48. H Peptide; and V containing 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: 49 L The peptide is ligated to a stalk moiety (S1) containing an Fc region provided herein. In some embodiments, a V containing 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 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: 49 L The peptide is linked to a stalk moiety (S1) which includes an Fc region further comprising a transmembrane domain. In some embodiments, the Fc region further comprising a transmembrane domain has the formula L1-Fc-L2-X1, where L1 is a linker provided herein or is absent, Fc is a modified Fc region provided herein, L2 is a linker provided herein or is absent, and X1 is a polypeptide comprising a transmembrane domain provided herein. As provided herein, X1 is a polypeptide of the formula ECD-T MThe polypeptide may also contain an ICD, where ECD is an extracellular domain or fragment thereof provided herein, or is absent, TM is a transmembrane domain provided herein, and ICD is an intracellular domain provided herein, or is absent. In some embodiments, the polypeptide contains 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 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: 49 L Peptides are ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes 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 sequence number 48. H Peptide; and V containing 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: 49 L The peptide is CD8 and / or CD28 ECD, T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes 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 sequence number 48. H Peptide; and V containing 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: 49 L The peptides are ECD, CD8 and / or CD28 T M, and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes 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 sequence number 48. H Peptide; and V containing 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: 49 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and are linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing the ICD. In some embodiments, V includes 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 sequence number 48. H Peptide; and V containing 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: 49 L Peptides are ECD, T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, the ICD including an env embedded motif provided herein. In some embodiments, a V containing 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 sequence number 48. H Peptide; and V containing 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: 49 L The peptide is CD8 and / or CD28 ECD, T M, and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, the ICD including an env embedded motif provided herein. In some embodiments, a V containing 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 sequence number 48. H Peptide; and V containing 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: 49 L The peptides are ECD, CD8 and / or CD28 T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) containing an ICD, the ICD including an env embedded motif provided herein. In some embodiments, a V containing 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 sequence number 48. H Peptide; and V containing 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: 49 L The peptides are CD8 and / or CD28 ECD, CD8 and / or CD28 T M , and linked to a stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD, the ICD includes an env embedded motif provided herein. In some embodiments, the sequence of sequence V includes an 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 sequence number 48 linked to the stalk portion (S1) including an Fc region (L1-Fc-L2-X1) including an ICD. HPeptide; and V containing 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: 49 L The peptide is immobilized on 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 embodiments, V H and V L The peptides bind to immune cells, such as those provided herein.

[0297] In some embodiments, V H Peptides and V L A polypeptide containing a peptide, V H The peptide contains 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; V L A polypeptide is provided, comprising a peptide whose sequence 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: 49.

[0298] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 49. However, V H Peptides and V LThe peptide comprises a light chain CDR having the sequence of SEQ ID NOs. 45-47; and / or a heavy chain CDR having the sequence of SEQ ID NOs. 42-44. In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 49. However, V H Peptides and V L The peptide comprises a light chain CDR1 having the sequence of SEQ ID NO: 45; a light chain CDR2 having the sequence of SEQ ID NO: 46; a light chain CDR3 having the sequence of SEQ ID NO: 47; and / or a heavy chain CDR1 having the sequence of SEQ ID NO: 42; a heavy chain CDR2 having the sequence of SEQ ID NO: 43; and a heavy chain CDR3 having the sequence of SEQ ID NO: 44. In some embodiments, V H or V L The CDR in the chain is as described in the combinations provided herein.

[0299] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 49. However, V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 45; LCDR2 having the sequence of SEQ ID NO: 46; and LCDR3 having the sequence of SEQ ID NO: 47.H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 42; HCDR2 having the sequence of SEQ ID NO: 43; and HCDR3 having the sequence of SEQ ID NO: 44.

[0300] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains 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; V L The peptide contains 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: 49. However, V L The peptide comprises LCDR1 having the sequence of SEQ ID NO: 45 and containing at most one conservative amino acid substitution; LCDR2 having the sequence of SEQ ID NO: 46 and containing at most one conservative amino acid substitution; and LCDR3 having the sequence of SEQ ID NO: 47 and containing at most one conservative amino acid substitution; V H The peptide comprises HCDR1 having the sequence of SEQ ID NO: 42 and containing at most one conservative amino acid substitution, HCDR2 having the sequence of SEQ ID NO: 43 and containing at most one conservative amino acid substitution, and HCDR3 having the sequence of SEQ ID NO: 44 and containing at most one conservative amino acid substitution.

[0301] In some embodiments, the polypeptide is V H Peptides and V L Contains peptides, V H The peptide contains the sequence of SEQ ID NO: 48, V L The peptide contains the sequence of SEQ ID NO: 49.

[0302] In some embodiments, the polypeptides provided herein bind to non-human primate CD8. In some embodiments, the polypeptides provided herein bind to human CD8.

[0303] As provided herein, different polypeptides (V H or V L ) may or may not be linked to a peptide linker, and may instead be a continuous sequence. In some embodiments, the peptide linker is (GGGGS) n The sequence (SEQ ID NO: 55) is included, where each n is independently 1 to 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 is V H -ZV L or V L -ZV H It can be expressed by the formula, where Z is a peptide linker. In some embodiments, Z is (GGGGS) n (Sequence ID 55) where each n is independently 1 to 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.

[0304] In some embodiments, formula V L -ZV H A polypeptide comprising the linked peptide represented by includes the heavy chain variable region described in SEQ ID NO: 48, linked to the light chain variable region described in SEQ ID NO: 49 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, via the peptide linker V H V connected to L The polypeptide containing has the sequence described below. NIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSS(Sequence No. 50).

[0305] In some embodiments, the polypeptide contains 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: 50. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 50. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 50. In some embodiments, the polypeptide described in SEQ ID NO: 50 is an antibody or its antigen-binding fragment. In some embodiments, the antibody 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.

[0306] In some embodiments, formula V H -ZV L A polypeptide comprising the linked peptide represented by includes a light chain variable region described in SEQ ID NO: 49 linked to the heavy chain variable region described in SEQ ID NO: 48 via the linker sequence GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 72). In some embodiments, via the peptide linker V L V connected to HThe polypeptide containing has the sequence described below. EVQLQQSGPELVKPGASVKISCKASRYTFTDYNLHWVKLSHEKSLEWIGFIYPYNGGTGYNQKFKNKAKLTVDYSSSTAYMELRSLTSVDAAVYYCARDHRYNEGVSFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSNIVLTQSPASLAVSLGQRATISCRASESVDGFGNSFMNWYQQKPGQSPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPYTFGGGTKLEIKR (Sequence ID 51).

[0307] In some embodiments, the polypeptide contains 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: 51. In some embodiments, the polypeptide contains a sequence that is at least 90% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide contains a sequence that is at least 95% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide contains a sequence that is at least 99% identical to the sequence of SEQ ID NO: 51. In some embodiments, the polypeptide contains the sequence described in SEQ ID NO: 51. In some embodiments, the polypeptide described in SEQ ID NO: 51 is an antibody or its antigen-binding fragment. In some embodiments, the antibody is an anti-CD8 antibody. In some embodiments, the anti-CD8 antibody binds to non-human primate CD8. In some embodiments...

Claims

1. a) Chimeric antigen receptors (CARs); and b) A polypeptide, i) Ligand-binding extracellular domain (ECD); ii) Intracellular domains that activate immune cells (ICDs); and iii) A particle or composition comprising one or more nucleic acid molecules encoding a polypeptide, which includes a transmembrane domain linking the ECD to the ICD.

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

3. The particle according to claim 2, wherein the hGCSF-R ICD includes the sequence of Sequence 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 thereto.

4. The particle according to claim 3, wherein the hGCSF-R ICD does not contain the sequence of sequence number 110.

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

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

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

10. The particle according to claim 8, wherein the ECD of the hGH-R includes 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 thereto.

11. The particle according to claim 1, wherein the ICD comprises an ICD of glycoprotein 130 ("gp130 ICD"), or a modified or fragment thereof.

12. The particle according to claim 1, wherein the ICD comprises a fragment of the GCSF-R ICD, and the GCSF-R ICD fragment does not contain the STAT3, SH3, and / or the SHC domain.

13. The particle according to claim 1, wherein the ICD comprises IL-2Rβ ICD, or a fragment or modified thereof.

14. The particle according to claim 1, wherein the ICD comprises IL-7Rα ICD, or a fragment or modified thereof.

15. The particle according to claim 1, wherein the ICD comprises an IL-2Rβ ICD, or a gp130 ICD linked to a fragment thereof, or a modified or fragment thereof.

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

17. The polypeptide described above, a) ECDs that are GCSFR-ECDs, or their variants; transmembrane domains that are GCSF-R transmembrane domains, or their variants; and immunoactivated ICDs that are GCSF-R ICDs, or their variants; b) ECDs that are hGH-R-ECD, or variants thereof; transmembrane domains that are GCSF-R transmembrane domains, or variants thereof; and immunoactivated ICDs that are GCSF-R ICD, or variants thereof; c) ECDs that are hGH-R-ECD, or variants thereof; transmembrane domains that are gp130 transmembrane domains, or variants thereof; and immunoactivating ICDs comprising gp130 ICD, or variants thereof and IL-2Rβ ICD, or variants thereof; or d) The particle according to claim 1, comprising an ECD which is hGH-R-ECD, or a variant thereof; a transmembrane domain which is a GCSF-R transmembrane domain, or a variant thereof; and an immunoactivating ICD which comprises a GCSF-R ICD, or a variant thereof and an IL-7Rα ICD, or a variant thereof.

18. The particle according to claim 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 any of the aforementioned modified versions.

19. The particle according to claim 18, wherein the modified product 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 respect to 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.

20. The particle according to claim 1, wherein the CAR includes an antigen-binding domain that binds to CD20.

21. The particle according to claim 20, wherein the CAR contains 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, 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.

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

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

24. i) Ligand-binding extracellular domain (ECD); ii) Intracellular domains that activate immune cells (ICDs); and iii) A nucleic acid molecule comprising an isolated nucleic acid molecule encoding a polypeptide including a transmembrane domain that links the ECD to the ICD.

25. A nucleic acid molecule according to claim 24, which encodes a polypeptide according to any one of claims 1 to 19.

26. The nucleic acid molecule according to claim 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 any of the above-mentioned variants.

27. The nucleic acid molecule according to claim 26, wherein the modified product 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 respect to 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.

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

29. A viral particle comprising a heterologous viral glycoprotein, a targeting moiety, and one or more nucleic acid molecules encoding one or more target heterologous molecules, The targeting moiety has the formula T-S １ The polypeptide comprises having T as the target-binding domain and S １ However, this is the stalk part, and A virus particle in which the nucleic acid molecule encoding the target heterologous molecule is the nucleic acid molecule described in any one of claims 24 to 28.

30. The target-binding domain (T) may be CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR alpha, TCR beta, TCR gamma, TCR delta, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3; a glycosylated CD43 epitope expressed in acute leukemia or lymphoma but not in hematopoietic progenitor cells; a glycosylated CD43 epitope expressed on non-hematopoietic cancers; A kinase anchor protein 4 (AKAP-4); adrenaline receptor β3 (ADRB3); AFP; anaplastic Lymphoma kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie2); autoantibody against desmoglein 1 (Dsg1); autoantibody against desmoglein 3 (Dsg3); 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); CC CTC binding factor (Zinc Finger Protein)-like (BORIS or Brother of the Regulator of lmprinted Sites); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32 (FCGR2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGLL; CD200R; CD276 / B7H3; CD300 molecule-like family member f (CD300LF); CDH1-CD324; CDH6; CDH17; CDH19;X chromosome open reading frame 61 (CXORF61); Claudin 6 (CLDN6); Claudin l8.2 (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 IB1 (CYP1B 1) DLL3; EBV-EBNA3c, EGF-bke module-containing mucin-like hormone receptor 2 (EMR2); elongation factor 2 mutated (ELF2M); ephrin B2; ephrin type A receptor 2 (EphA2); epidermal growth factor receptor (EGFR); epidermal growth factor receptor variant III (EGFRviii); epidermal cell adhesion molecule (EPCAM); ERG; ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML); IgA receptor Fc fragment (FCAR or CD89); Fc receptor-like 5 5, FCRL5); Fibroblast-activating protein α (FAP); FITC; Fms-like tyrosine kinase 3 (FLT3); Folate receptor alpha (Fra or FR1); Folate receptor β (FRb); Follicle-stimulating hormone receptor (FSHR); Fos-related antigen 1; Fucosyl-GM1; G protein-coupled receptor class C group 5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; Ganglioside G2 (GD2); Ganglioside GD3 (αNeu 5 Ac(2-8)αNeu 5 Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer);Ganglioside GM3 (αNeu 5 Ac(2-3) bDClarp(1-4) bDClcp(1-1)Cer); GD3; GFRalpha4; Glycoprotein 100 (gplOO); Glypican-3 (GPC3); Gonadotropin hormone receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylyl cyclase C (GCC); Heat shock protein 70-2 mutated (mut hsp70-2); Hepatitis A virus cellular receptor 1 1. HAVCR1); hexasaccharide portion of globeH glycoceramide (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 papilloma virus E6 (HPV E6); human papilloma virus E7 (HPV E7); human telomerase reverse transcriptase (hTERT); IgE; IL13Ra2; Ill lRa; immunoglobulin λ-like polypeptide 1 (IGLL1); influenza A hemagglutinin (HA); insulin-like growth factor 1 receptor (IGF-I receptor); interleukin-11 receptor alpha (IL-11Ra); interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2); intestinal carboxylesterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; Regmine; leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); luteinizing hormone receptor (LHR); Lewis (Y) antigen;Lewis Ag; Livl; K9 locus (LY6K); low conductance chloride channel; lymphocyte antigen 6 complex; lymphocyte antigen 75 (LY75); lymphocyte-specific protein tyrosine kinase (LCK); mammary gland differentiation antigen (NY-BR-1); melanoma antigen recognized by T cell 1 (Melanoa or MART 1); melanoma-associated antigen 1 (MAGE-A1); melanoma cancer testis antigen-1 (MAD-CT-1); melanoma cancer testis antigen-2 (MAD-CT-2); melanoma inhibitor of apoptosis (ML-IAP); mesothelin; MPL; mucin 1 cell surface-bound (MUC1); N-acetylglucosaminyltransferase V (NA17); nectin-4; neural cell adhesion molecule (NCAM); NKG2D; NYBR1; O-acetyl-GD2 ganglioside (OacGD2); olfactory receptor 51E2 (OR51E2); oncogene fusion protein consisting of a cleavage cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Ab1) (bcr-ab1); P53 variant; paired box protein Pax-3 (PAX3); paired box protein Pax-5 (PAX5); pannexin 3 (pannexin 3. PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor beta (PDGFR-beta); polysialic acid; proacrosin-binding protein sp32 (OY-TES1); prostase; prostate cancer tumor antigen-1 (PCT A-1 or galectin 8); prostate stem cell antigen (PSCA);Prostate-specific membrane antigen (PSMA); prostatic acid phosphatase (PAP); prostain; protease serine 21 (testisin or PRSS21); proteasome (prosome, macropain) subunit beta 9 (LMP2); PTK7; Ras G12V; Ras homolog family member C (RhoC); rat sarcoma (Ras) mutant; advanced glycation end product receptor (RAGE-1); receptor tyrosine kinase-like orphan receptor 1 (ROR1); receptor tyrosine-protein kinase ERBB2 or Her-22 / neu; renal ubiquitous 1 (RU1); renal ubiquitous 2 (renal ubiquitous 2, RU2); sarcoma translocation breakpoint; serine 2 (TMPRSS2) ETS fusion gene; sialyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialyl Lewis antigen); sperm protein 17 (SPA17); squamous cell carcinoma antigen 3 recognized by T cells (SART3); stage-specific embryonic antigen-4 (SSEA-4); STEAP1; Survivin; synovial sarcoma, X breakpoint 2 (SSX2); TCR gamma alternative reading frame protein (TCR Gamma Alternate Reading Frame Protein (TARP); TCR-beta-1 chain; TCR-beta-2 chain; TCR-delta chain; TCR-gamma chain; TCR-gamma-delta; telomerase; TGFbetaR2; 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-related (TEM7R); tumor protein p53 (p53); tumor-associated glycoprotein 72 (TAG72); tyrosinase; tyrosinase-related protein 2 (TRP-2); uroplakin 2 (UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein A viral particle according to claim 29, which binds to protein (WT1); or to member 1A (XAGE1) of the X antigen family.

31. The virus particle according to claim 30, wherein the target-binding domain (T) binds to CD7.

32. The virus particle according to claim 31, wherein the target binding domain (T) that binds to CD7 comprises a polypeptide having at least 90% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, a sequence having at least 95% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, a sequence having at least 99% sequence identity with SEQ ID NO: 38 or SEQ ID NO: 39, or a sequence described in SEQ ID NO: 38 or SEQ ID NO:

39.

33. The virus particle according to any one of claims 29 to 32, wherein the heterologous viral glycoprotein is a VSV-G polypeptide.

34. The virus particle according to claim 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.

35. A viral particle comprising a heterologous viral glycoprotein, a targeting moiety, and a nucleic acid molecule encoding the target heterologous molecule, 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; The targeted portion includes an amino acid sequence having at least 90% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, at least 95% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, at least 99% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, or at least 100% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127; and The nucleic acid molecule encoding the target heterologous molecule, Chimeric antigen receptor, and i) Ligand-binding extracellular domain (ECD); ii) Intracellular domains that activate immune cells (ICDs); and iii) A virus particle encoding a polypeptide including a transmembrane domain that links the ECD to the ICD.

36. The virus particle according to claim 35, wherein the CAR includes 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.

37. The virus particle according to claim 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 any of the aforementioned variants.

38. The virus particle according to any one of claims 35 to 37, wherein the chimeric antigen receptor and the polypeptide are encoded by separate nucleic acid molecules.

39. The virus particle according to any one of claims 35 to 37, wherein the chimeric antigen receptor and the polypeptide are encoded by the same nucleic acid molecule.

40. The virus particle according to claim 39, wherein the chimeric antigen receptor and the polypeptide are separated by a sequence encoding a cleavable peptide, such as a 2A peptide.

41. A method for infecting, transfecting, or transfecting cells, wherein the method includes contacting the cells with particles according to any one of claims 1 to 23 or with viral particles according to any one of claims 29 to 40.

42. A method for delivering one or more target heterologous molecules to a cell, wherein the method comprises contacting the cell with a particle according to any one of claims 1 to 23 or a viral particle according to any one of claims 29 to 40, wherein the viral particle comprises a nucleic acid molecule encoding the target heterologous molecule.

43. The method according to claim 41 or 42, further comprising administering to the cells or subject one or more compounds that bind to the ECD of the polypeptide encoded by one or more nucleic acid molecules, such as but not limited to somatropin, G-CSF, lenograstim (granocyte), filgrastim (newpogen, salzio, nivestim, acofil), long-acting (pegylated) filgrastim (pegylated filgrastim, neurusta, permeg, ziextenko), or lipezilgrastim (lonkex).

44. A method for treating a disease or disorder in a subject, wherein the method comprises administering to the subject a particle according to any one of claims 1 to 23 or a viral particle according to any one of claims 29 to 40, wherein the viral particle comprises a nucleic acid molecule encoding a target heterogeneous molecule for treating the disease or disorder, and the nucleic acid molecule encoding the target heterogeneous molecule is the nucleic acid molecule according to any one of claims 24 to 28.

45. A method for treating a disease or disorder in a subject, wherein the method is i) Administering the particles described in any one of claims 1 to 23 or the virus particles described in any one of claims 29 to 40 to the subject, 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 disorder, The virus particle comprises one or more nucleic acid molecules encoding one or more target heterogeneous molecules for treating the disease or disorder, A method wherein the one or more nucleic acid molecules encoding one or more target heterogeneous molecules are the one or more nucleic acid molecules described in any one of claims 24 to 28.

46. A method for treating a disease or disorder in a subject, wherein the method is i) Administering the particles described in any one of claims 1 to 23 or the virus particles described in any one of claims 29 to 40 to the subject, 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 disorder, The virus particle comprises one or more nucleic acid molecules encoding one or more target heterogeneous molecules for treating the disease or disorder, A method wherein the one or more nucleic acid molecules encoding one or more target heterogeneous molecules are the one or more nucleic acid molecules described in any one of claims 24 to 28.

47. A method for treating a disease or disorder in a subject, wherein the method is i) Administering the virus particles described in any one of claims 29 to 40 to the subject, 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 disorder, The virus particle comprises a nucleic acid molecule encoding a heterogeneous molecule intended for treating the disease or disorder. The nucleic acid molecule or a plurality of nucleic acid molecules encoding the target heterologous molecule are chimeric antigen receptors, and i) Ligand-binding extracellular domain (ECD); ii) Intracellular domains that activate immune cells (ICDs); and iii) A method for encoding a polypeptide comprising a transmembrane domain that links the ECD to the ICD.

48. The method according to claim 47, wherein the CAR includes 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.

49. The method according to claim 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 any of the above-mentioned variants.

50. A method for delivering one or more heterologous molecules to target cells, wherein the method comprises contacting the cells with particles according to any one of claims 1 to 23 or with a viral vector or particles according to any one of claims 29 to 40, wherein the particles comprise one or more nucleic acid molecules encoding the one or more heterologous molecules.

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