Antigen-binding domains and methods of use thereof

Abstract

Provided herein are antibodies and antigen binding fragments thereof specific for V-set Immunoglobulin domain containing 2 (VSIG2). Also provided herein are cells, nucleic acids, vectors, compositions, and methods directed to antibodies or antigen-binding domains thereof specific for VSIG2.

Description

Invention title: ANTIGEN-BINDING DOMAINS AND METHODS OF USE THEREOF. Provided herein are antibodies and antigen-binding fragments thereof specific for the V-set Immunoglobulin domain containing 2 (VSIG2). Also provided herein are cells, nucleic acids, vectors, compositions, and methods directed to antibodies or antigen-binding domains thereof specific for VSIG2. Fig. 1 Abstract

Claims

CLAIMSWhat is claimed is:

1. An isolated antibody or antigen binding fragment thereof that specifically binds to human V- set Immunoglobulin domain containing 2 (VSIG2) comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH comprises a VH complementarity region 1 (CDRH1) having the amino acid sequence of SEQ ID NO: 1 or 2, and a VH complementarity region 2 (CDRH2) having the amino acid sequence of SEQ ID NO: 3 or 4; wherein the VL comprises a VL complementarity region 1 (CDRL1) having the amino acid sequence of SEQ ID NO: 6, and a VL complementarity region 2 (CDRL2) having the amino acid sequence of SEQ ID NO: 7; and wherein:(i) the VH comprises a VH complementarity region 3 (CDRH3) having the amino acid sequence of SEQ ID NO: 5, and the VL comprises a VL complementarity region 3 (CDRL3) having the amino acid sequence of SEQ ID NO: 9, or(ii) the VH comprises a VH complementarity region 3 (CDRH3) having the amino acid sequence of any one of SEQ ID NO: 67-87, and the VL comprises a VL complementarity region 3 (CDRL3) having the amino acid sequence of SEQ ID NO: 8 or 9, optionally wherein the VH has an amino acid sequence selected from the group consisting of SEQ ID NO: 16 and 88-107, optionally wherein the VL has an amino acid sequence of SEQ ID NO: 14 or SEQ ID NO: 15.

2. The antibody or antigen binding fragment thereof of claim 1, wherein:A) the CDRH3 has the amino acid sequence of SEQ ID NO: 5, and the CDRL3 has the amino acid sequence of SEQ ID NO: 9; orB) the CDRH3 has the amino acid sequence of SEQ ID NO: 67, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orC) the CDRH3 has the amino acid sequence of SEQ ID NO: 68, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orD) the CDRH3 has the amino acid sequence of SEQ ID NO: 67, and the CDRL3 has the amino acid sequence of SEQ ID NO: 9; orE) the CDRH3 has the amino acid sequence of SEQ ID NO: 68, and the CDRL3 has the amino acid sequence of SEQ ID NO: 9; orF) the CDRH3 has the amino acid sequence of SEQ ID NO: 69, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orG) the CDRH3 has the amino acid sequence of SEQ ID NO: 69, and the CDRL3 has the amino acid sequence of SEQ ID NO: 9; orH) the CDRH3 has the amino acid sequence of SEQ ID NO: 70, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orI) the CDRH3 has the amino acid sequence of SEQ ID NO: 71, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orJ) the CDRH3 has the amino acid sequence of SEQ ID NO: 72, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orK) the CDRH3 has the amino acid sequence of SEQ ID NO: 73, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orL) the CDRH3 has the amino acid sequence of SEQ ID NO: 74, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orM) the CDRH3 has the amino acid sequence of SEQ ID NO: 75, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orN) the CDRH3 has the amino acid sequence of SEQ ID NO: 76, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orO) the CDRH3 has the amino acid sequence of SEQ ID NO: 77, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orP) the CDRH3 has the amino acid sequence of SEQ ID NO: 78, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orQ) the CDRH3 has the amino acid sequence of SEQ ID NO: 79, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orR) the CDRH3 has the amino acid sequence of SEQ ID NO: 80, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orS) the CDRH3 has the amino acid sequence of SEQ ID NO: 81, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orT) the CDRH3 has the amino acid sequence of SEQ ID NO: 82, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orU) the CDRH3 has the amino acid sequence of SEQ ID NO: 83, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orV) the CDRH3 has the amino acid sequence of SEQ ID NO: 84, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orW) the CDRH3 has the amino acid sequence of SEQ ID NO: 85, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orX) the CDRH3 has the amino acid sequence of SEQ ID NO: 86, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8; orY) the CDRH3 has the amino acid sequence of SEQ ID NO: 87, and the CDRL3 has the amino acid sequence of SEQ ID NO: 8.

3. The antibody or antigen binding fragment thereof of claim 1 or claim 2, wherein the antibody or antigen binding fragment thereof is an antigen binding fragment, optionally wherein the antigen binding fragment comprises a F(ab) fragment, a F(ab’) fragment, or a single chain variable fragment (scFv), optionally wherein the antigen binding fragment comprises a single chain variable fragment (scFv), optionally wherein the scFv comprises a VH and a VL separated by a peptide linker, optionally wherein the antigen-binding domain comprises the structure VH- L-VL or VL-L-VH, wherein VH is the heavy chain variable domain, L is the peptide linker, and VL is the light chain variable domain, optionally wherein the peptide linker comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 21-37, optionally wherein the scFv comprises an amino acid sequence selected from the group consisting of: SEQ ID Nos: 108-132.

4. A chimeric protein comprising an antibody or antigen binding fragment thereof of any one of claims 1-3 and a heterologous molecule or moiety, optionally wherein the chimeric protein isan antibody-drug conjugate, and wherein the heterologous molecule or moiety comprises a therapeutic agent, optionally wherein the chimeric protein is a chimeric antigen receptor (CAR), and wherein the heterologous molecule or moiety comprises a polypeptide selected from the group consisting of: a transmembrane domain, one or more intracellular signaling domains, a hinge domain, a spacer region, one or more peptide linkers, and combinations thereof, optionally wherein the CAR is an inhibitory CAR comprising one or more intracellular inhibitory domains that inhibit an immune response, optionally wherein the one or more intracellular inhibitory domains comprise an ICD derived from PD-1, CTLA4, TIGIT, BTLA, LIR1 (LILRB1), TIM3, KIR3DL1, NKG2A , LAG3, LAIR1, SIRPa, KIR2DL1, KIR2DL2, KIR2DL3, KIR3DL2, KLRG-1, CEACAM1, LIR2, LIR3, LIR5, SIGLEC-2, SIGLEC-10, PECAM-1, CD72, IRTA2, IRTA4, NKIR, TLT1, PCDHGC3, MPZL1, FCGR2B, SIGLEC-6, MPIG6B, SIGLEC-12, LIR8, IRTA1, KIR2DL4, KIR2DL5, SIGLEC-7, or FCRH3, optionally wherein the intracellular inhibitory domain comprises the amino acid sequence VRIRQKKAQGSTSSTRLHEPEKNAREITQDTNDITYADLNLPKGKKPAPQAAEPNNHTE YASIQTSPQPASEDTLTYADLDMVHLNRTPKQPAPKPEPSFSEYASVQVPRK (SEQ ID NO: 139) or an amino acid sequence with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to one or more of the amino acid sequenceVRIRQKKAQGSTSSTRLHEPEKNAREITQDTNDITYADLNLPKGKKPAPQAAEPNNHTE YASIQTSPQPASEDTLTYADLDMVHLNRTPKQPAPKPEPSFSEYASVQVPRK (SEQ ID NO: 139), optionally wherein the intracellular inhibitory domain further comprises an enzymatic inhibitory domain and / or an additional intracellular inhibitory co-signaling domain, optionally wherein the CAR comprises a spacer region between the antigen-binding domain and the transmembrane domain, optionally wherein the spacer region has an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-52.

5. An engineered expression system comprising: a. a first nucleic acid sequence encoding a first CAR, wherein the first CAR comprises: i. a first extracellular antigen-binding domain that binds an antigen selected from the group consisting of CEACAM5, CEA, CEACAM1, and CEACAM6; ii. a first transmembrane domain; andiii. one or more intracellular signaling domains; and b. a second nucleic acid sequence encoding a second CAR, wherein the second CAR comprises the antibody or antigen binding fragment of any one of claims 1 - 3 or the chimeric protein of claim 4.

6. The engineered expression system of claim 5, wherein the first CAR comprises a first spacer between the first extracellular antigen-binding domain and the first transmembrane domain, optionally wherein the first spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-52, optionally wherein the first spacer comprises the amino acid sequence of SEQ ID NO: 5, and / or wherein the second CAR comprises a second spacer between the second extracellular antigen-binding domain and the second transmembrane domain, optionally wherein the second spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-52, optionally wherein the second spacer comprises the amino acid sequence of SEQ ID NO: 50.

7. The engineered expression system of claim 5 or claim 6, wherein the one or more intracellular signaling domains of the first CAR are selected from the group consisting of: a CD3zeta-chain intracellular signaling domain, a CD3epsilon-chain intracellular signaling domain, a CD97 intracellular signaling domain, a CD1 la-CD18 intracellular signaling domain, a CD2 intracellular signaling domain, an ICOS intracellular signaling domain, a CD27 intracellular signaling domain, a CD 154 intracellular signaling domain, a CD8 intracellular signaling domain, an 0X40 intracellular signaling domain, a 4- IBB intracellular signaling domain, a CD28 intracellular signaling domain, a ZAP40 intracellular signaling domain, a CD30 intracellular signaling domain, a GITR intracellular signaling domain, an HVEM intracellular signaling domain, a DAP 10 intracellular signaling domain, a DAP 12 intracellular signaling domain, a MyD88 intracellular signaling domain, a 2B4 intracellular signaling domain, an NKp46 intracellular signaling domain, an NKp30 intracellular signaling domain, an NKp44 intracellular signaling domain, an NKG2D intracellular signaling domain, a CD226 intracellular signaling domain, and a CD 160 intracellular signaling domain, optionally wherein the first CAR comprises a CD28 intracellular signaling domain and a CD3zeta-chain intracellular signaling domain; and / or optionally wherein the first transmembrane domain are selected from the group consisting of: a CD8 transmembrane domain, a CD28 transmembrane domain, a CD25 transmembrane domain, a CD7 transmembrane domain, a CD3zeta-chain transmembrane domain, a CD4 transmembrane domain, a 4- IBB transmembrane domain, an 0X40 transmembrane domain, an ICOS transmembrane domain, a CTLA-4 transmembrane domain, aLAX transmembrane domain, a LAT transmembrane domain, a PD-1 transmembrane domain, a LAG-3 transmembrane domain, a TIM3 transmembrane domain, a KIR3DS1 transmembrane domain, a KIR3DL1 transmembrane domain, an NKG2D transmembrane domain, an NKG2A transmembrane domain, a TIGIT transmembrane domain, a 2B4 transmembrane domain, and a BTLA transmembrane domain, optionally wherein the first CAR comprises a CD28 transmembrane domain.

8. The engineered expression system of any one of claims 5-7, wherein the first and second nucleic acid sequences are comprised within a single expression vector; or wherein the first nucleic acid sequence is comprised within a first expression vector and the second nucleic acid sequence is comprised within a second expression vector.

9. The engineered expression system of any one of claims 5-8, wherein the first antigenbinding domain binds to any one of CEACAM5, CEACAM6, or CEACAM1, wherein the first antigen-binding domain comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), optionally wherein: i) the VH comprises a VH complementarity region 1 (CDRH1) , a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of an hMN14 VH, and wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of an hMN14 VL, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an hMN14 VH, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an hMN14 VL, optionally wherein the VH comprises the amino acid sequence of an hMN 14 VH, and the VL comprises the amino acid sequence of an hMN 14 VL, optionally wherein the first antigen-binding domain comprises a heavy chain variable domain (VH) and a light chain variable domain (VL); or ii) the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of a BW431 / 26 VH; optionally wherein the VL comprises a VLcomplementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of a BW431 / 26 VL, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of a BW431 / 26 VH, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of a BW431 / 26 VL, optionally wherein the VH comprises the amino acid sequence of a BW431 / 26 VH, and the VL comprises the amino acid sequence of a BW431 / 26 VL; or iii) the first antigen-binding domain comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of an A5B7 VH; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of an A5B7 VL, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an A5B7 VH, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an A5B7 VL, optionally wherein the VH comprises the amino acid sequence of an A5B7 VH, and the VL comprises the amino acid sequence of an A5B7 VL, iv) the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of an MFE23 VH; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of an MFE23 VL, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VHcomprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an MFE23 VH, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an MFE23 VL, optionally wherein the VH comprises the amino acid sequence of an MFE23 VH, and the VL comprises the amino acid sequence of an MFE23 VH; or v) the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of an hMFE23 VH; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of an hMFE23 VL, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an hMFE23 VH, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an hMFE23 VL, optionally wherein the VH comprises the amino acid sequence of an hMFE23 VH, and the VL comprises the amino acid sequence of an hMFE23 VL; or vi) the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of an FM4 VH; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of an FM4 VL, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an FM4 VH, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an FM4 VL,optionally wherein the VH comprises the amino acid sequence of an FM4 VH, and the VL comprises the amino acid sequence of an FM4 VL; or vii) the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of a cibisatamab HC; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of a cibisatamab LC, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of the VH of a cibisatamab HC, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of the VL of a cibisatamab LC, optionally wherein the VH comprises the amino acid sequence of the VH of a cibisatamab HC and the VL comprises the amino acid sequence of the VL of a cibisatamab LC; or viii) the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of a tusamitamab HC; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of a tusamitamab LC, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of the VH of a tusamitamab HC, and the LC comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of the VL of a tusamitamab LC, optionally wherein the HC comprises the amino acid sequence of the VH of a tusamitamab HC, and the VL comprises the amino acid sequence of the VL of a tusamitamab LC; or ix) the first antigen-binding domain comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises a VHcomplementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of an MRG1 VH; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of an MRG1 VL, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an MRG1 VH, and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of an MRG1 VL, optionally wherein wherein the VH comprises the amino acid sequence of an MRG1 VH, and the VL comprises the amino acid sequence of an MRG1 VL; or x) the first antigen-binding domain comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises a VH complementarity region 1 (CDRH1), a VH complementarity region 2 (CDRH2), and a VH complementarity region 3 (CDRH3) of a tinurilimab HC; wherein the VL comprises a VL complementarity region 1 (CDRL1), a VL complementarity region 2 (CDRL2), and a VL complementarity region 3 (CDRL3) of a tinurilimab LC, and wherein the antibody or antigen binding fragment thereof is humanized, optionally wherein the VH comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of the VH of a a tinurilimab HC and the VL comprises an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of the VL of a tinurilimab LC, optionally wherein the VH comprises the amino acid sequence of the VH of a tinurilimab HC and the VL comprises the amino acid sequence of the VL of a tinurilimab LC.

10. The engineered expression system of any one of claims 5-9, further comprising: a. A fourth nucleotide sequence encoding a first cytokine; andb. A fifth nucleotide sequence encoding a second cytokine.

11. The engineered expression system of any one of claims 5-10, wherein at least one of the first and the second cytokines is a controlled release cytokine, optionally wherein the controlled release cytokine has the formula:S - C - MT or MT - C - S whereinS comprises a secretable effector molecule;C comprises a protease cleavage site; andMT comprises a cell membrane tethering domain. optionally wherein the protease cleavage site is cleaved by ADAM10 and / or ADAM17, optionally wherein the protease cleavage site comprises the amino acid sequence of PRAEALKGG or VTPEPIFSLI, optionally wherein the cell membrane tethering domain comprises a transmembrane domain selected from the group consisting of: PDGFR-beta, CD8, CD28, CD3zeta-chain, CD4, 4- IBB, 0X40, ICOS, CTLA-4, PD-1, LAG-3, 2B4, LNGFR, NKG2D, EpoR, TNFR2, LIR1, B7-1, and BTLA, optionally wherein the cell membrane tethering domain comprises a B7-1 transmembrane domain comprising the B7-1 transmembrane domain amino acid sequence set forth in Table 14.

12. The engineered expression system of any one of claims 5-11, wherein the first cytokine is IL15, optionally wherein the IL15 comprises the amino acid sequence of IL15 set forth in Table 10, optionally wherein the IL15 is controlled-release IL15 (crIL15).

13. The engineered expression system of any one of claims 5-12, wherein the second cytokine is IL21, optionally wherein the IL21 comprises the amino acid sequence set forth in Table 10, optionally wherein the IL21 is controlled-release IL21 (crIL21).

14. An engineered nucleic acid encoding the antibody or antigen binding fragment of any one of claims 1-3, the chimeric protein of claim 4, or the engineered expression system of any one of claims 5-13, or optionally a vector comprising the engineered nucleic acid.

15. A composition comprising the antibody or antigen binding fragment thereof of any one of claims 1-3 or the chimeric protein of claim 4, the engineered nucleic acid of claim 14, or theengineered expression system of any one of claims 5-13, and a pharmaceutically acceptable carrier, pharmaceutically acceptable excipient, or a combination thereof.

16. An isolated cell or population of engineered cells comprising the engineered nucleic acid of claim 14, the composition of claim 15, the engineered expression system of any one of claims 5- 13, the antigen binding fragment of any one of claims 1-3, the chimeric protein of claim 4, optionally wherein the cell or population of cells is selected from the group consisting of: a T cell, a CD8+ T cell, a CD4+ T cell, a gamma-delta T cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell, a viral-specific T cell, a Natural Killer T (NKT) cell, a Natural Killer (NK) cell, a B cell, a tumor-infiltrating lymphocyte (TIL), an innate lymphoid cell, a mast cell, an eosinophil, a basophil, a neutrophil, a myeloid cell, a macrophage, a monocyte, a dendritic cell, an erythrocyte, a platelet cell, a human embryonic stem cell (ESC), an ESC-derived cell, a pluripotent stem cell, a mesenchymal stromal cell (MSC), an induced pluripotent stem cell (iPSC), and an iPSC-derived cell, optionally wherein the cell is autologous or wherein the cell is allogeneic, optionally wherein the cell or population of cells further comprises one or more tumor-targeting chimeric receptors expressed on the cell surface, optionally wherein at least one of the one or more tumor-targeting chimeric receptors is a chimeric antigen receptor (CAR) or an engineered T cell receptor.

17. A pharmaceutical composition comprising: a. an effective amount of the cell or population of engineered cells of claim 16 and a pharmaceutically acceptable carrier, pharmaceutically acceptable excipient, or a combination thereof; or b. an effective amount of genetically modified cells expressing the antigen binding fragment of any one of claims 1-3 or the chimeric protein of claim 4 and a pharmaceutically acceptable carrier, pharmaceutically acceptable excipient, or a combination thereof, optionally wherein the pharmaceutical composition is for treating and / or preventing a tumor.

18. A method of inhibiting a cell-mediated immune response to a normal cell in a subject, the method comprising administering to a subject a. a therapeutically effective dose of the composition of claim 15,b. any of the cells of claim 16, optionally wherein the isolated cell or population of cells express the chimeric protein comprising the inhibitory CAR of claim 4, c. the composition of claim 17; optionally wherein the method further comprises stimulating a cell-mediated immune response to a tumor cell in the subject, and wherein the isolated cell or population of cells further comprises one or more tumor-targeting chimeric receptors expressed on the cell surface, optionally wherein at least one of the one or more tumor-targeting chimeric receptors is a chimeric antigen receptor (CAR) or an engineered T cell receptor, optionally wherein the normal cell comprises a human VSIG2, optionally wherein the human VSIG2 is expressed on a surface of the normal cell, optionally wherein the normal cell is a healthy (e.g., non-tumor) epithelial cell.

19. A method of treating a subject having a tumor, the method comprising administering a therapeutically effective dose of the composition of claim 15, or any of the cells of claim 16, or the composition of claim 17.

20. A kit for treating and / or preventing a tumor, comprising: a. the chimeric protein of claim 4, optionally wherein the kit further comprises written instructions for using the chimeric protein for producing one or more antigen-specific cells for treating and / or preventing a tumor in a subject. b. the cell or population of cells of claim 16, optionally wherein the kit further comprises written instructions for using the cell for treating and / or preventing a tumor in a subject c. the vector of claim 14, optionally wherein the kit further comprises written instructions for using the vector for producing one or more antigen-specific cells for treating and / or preventing a tumor in a subject; d. the composition of claim 15 or claim 17, optionally wherein the kit further comprises written instructions for using the composition for treating and / or preventing a tumor in a subject;e. the engineered nucleic acid of claim 14, optionally wherein the kit further comprises written instructions for using the nucleic acid for producing one or more antigen-specific cells for treating and / or preventing a tumor in a subject.

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