Chimeric antigen receptors and uses thereof

Abstract

The disclosure relates to new chimeric antigen receptors (CARs), new CAR-T cells, new methods for manufacturing new CARs and new CAR-T cells, as well as new methods for treating diseases, including viral disorders.

Description

ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION CHIMERIC ANTIGEN RECEPTORS AND USES THEREOF CROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of U.S. Provisional Application No.63 / 665,232, which was filed June 27, 2024, is titled “CHIMERIC ANTIGEN RECEPTORS AND USES THEREOF,” and is incorporated herein by reference in its entirety. SEQUENCE LISTING

[0002] The electronic sequence listing filed herewith, titled “STFD-014-PCT_SL.xml,” created June 27, 2025, and having a file size of 486,360 bytes is incorporated herein by reference in its entirety. BACKGROUND

[0003] Many patients with viral infections are incurable with standard therapy. In addition, traditional treatment options often have serious side effects.

[0004] Recent developments using chimeric antigen receptor (CAR) modified autologous T cell (CART) therapy, which relies on redirecting T cells to a suitable cell-surface molecule on cancer cells such as B cell malignancies, show promising results in harnessing the power of the immune system to treat B cell malignancies and other cancers (see, e.g., Sadelain et al., Cancer Discovery 3:388-398 (2013)). The clinical results of the murine derived CART19 (i.e., “CTL019”) have shown promise in establishing complete remissions in patients suffering with CLL as well as in childhood ALL (see, e.g., Kalos et al., Sci Transl Med 3:95ra73 (2011), Porter et al., NEJM 365:725-733 (2011), Grupp et al., NEJM 368:1509-1518 (2013)). Besides the ability for the chimeric antigen receptor on the genetically modified T cells to recognize and destroy the targeted cells, a successful therapeutic T cell therapy needs to have the ability to proliferate and persist over time, in order to survey for relapse or reinfection. The variable quality of T cells, resulting from anergy, suppression, or exhaustion, will have effects on CAR- transformed T cells’ performance, over which skilled practitioners have limited control at this time. To be effective, CAR-transformed patient T cells need to persist and maintain the ability to proliferate in response to the cognate antigen. SUMMARY OF EMBODIMENTS

[0005] The invention provides new chimeric antigen receptors (CARs) , new CAR-T cells, new methods for manufacturing new CARs and new CAR-T cells, as well as new methods for treating diseases, including viral disorders. In some embodiments, the viral disorder is a disease 1ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION caused by a virus. In some embodiments, the viral disorder is a viral infection. In some embodiments, the viral disorder is a cancer caused by a virus. In some embodiments, the viral disorder comprises a cell expressing one or more of an EBV antigen, an SL9 antigen, a Nef antigen, and a SARS antigen. In some embodiments, the viral disorder comprises a cell expressing one or more of (1) an EBV antigen and an HLA molecule comprising HLA-A*02, or a functional variant thereof, (2) an SL9 antigen and an HLA molecule comprising HLA- A*02, or a functional variant thereof, (3) a Nef antigen and an HLA molecule comprising HLA- A*24, or a functional variant thereof, or (4) a SARS antigen and an HLA molecule comprising HLA-B*08, or a functional variant thereof. In some embodiments, the viral disorder is one differentiated by having EBV-A*02 as a biomarker therefor. In some ebmodiments, the viral disorder caused by Epstein through Barr virus. In some ebmodiments, the viral disorder is a cancer caused by Epstein through Barr virus. In some ebmodiments, the viral disorder is mononucleosis. In some embodiments, the viral disorder is one differentiated by having SL9- A02 as a biomarker therefor. In some embodiments, the viral disorder is one differentiated by having Nef_A*24 as a biomarker therefor. In some embodiments, the viral disorder is infection with human immunodeficiency virus 1 (HIV-1). In some embodiments, the viral disorder is acquired immune deficiency syndrome (AIDS). In some embodiments, the viral disorder is one differentiated by having SARS-B*08 as a biomarker therefor. In some embodiment, the viral disorder is infection with SARS-CoV2. In some embodiment, the viral disorder is COVID-19.

[0006] In some embodiments, the disclosure relates to a method of treating a viral disorder comprising administering to a patient in need thereof a pharmaceutical composition comprising a therapeutically effective amount of cells comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR) molecule that binds to cells expressing EBV, SL9, Nef, and SARS antigens. In some embodiments, the CAR molecule binds to cell expressing EBV and an HLA molecule comprising HLA-A*02:01, or a function variant thereof. In some embodiments, the CAR molecule binds to cell expressing SL9 and an HLA molecule comprising HLA-A*02 or a function variant thereof. In some embodiments, the CAR molecule binds to cell expressing Nef and an HLA molecule comprising HLA-A*24 or a function variant thereof. In some embodiments, the CAR molecule binds to cell expressing SARS and an HLA molecule comprising HLA-B*08 or a function variant thereof.

[0007] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ 2ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO:ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50).

[0008] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 4ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100).

[0009] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), 5ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150).

[0010] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), 6ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200).

[0011] In some embodiment, the first nucleic acid sequence encodes a peptide comprising an amino acid sequence with at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of one of SEQ ID NOS: 1 through 200.

[0012] In some embodiments, the first nucleic acid sequence encodes a peptide comprising GTHDKCENPKX1X2X3X4X5X6X7X8QNX9NVVX10TNKELEDIYDESN (SEQ ID NO:202), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 202, wherein X1X2X3X4X5X6 X7X8 is one of SEQ ID NOS: 1 through 200 or a functional variant thereof comprising about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200, and X9 and X10 are any amino acid. In some embodiments, the nucleic acid molecule further comprises a second nucleic acid sequence encoding a peptide comprising KEETKEVLKKFKEKVNQFX11EHAFDIINKYGDKEIX12NMMX13MLLWRVX14FRSFRI DANNVELIKFNIRVLDWIMAEADNDLSYFISQ (SEQ ID NO:203), or a functional variant 7ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION thereof comprising from about 75% sequence identity to SEQ ID NO: 203, wherein X11, X12, X13, and X14are any amino acid. In some embodiments, the nucleic acid molecule comprises the first nucleic acid sequence and a third nucleic acid sequence encoding a single chain antibody or antibody fragment that binds to a cell expressing CD3, the first nucleic acid sequence and the third nucleic acid sequence together encode a chimeric peptide comprising a first domain and a second domain, the first domain comprising the peptide encoded by the first nucleic acid sequence, and the second domain comprising the single chain antibody or antibody fragment that binds to a cell expressing CD3. In some embodiments, the nucleic acid molecule comprising the first and third nucleic acid sequences further comprises the second nucleic acid sequence, and the first domain comprises the peptide encoded by the first nucleic acid sequence and the peptide encoded by the second nucleic acid sequence.

[0013] In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is about 3x104cells / kg body weight for patients with less than or equal to 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is about 1.5 x 106cells for patients with more than 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is from about 3.0 x 105to 1.5 x l06cells. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is about 10 x l04cells / kg body weight for patients with less than or equal to about 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is from about 5 x106cells for patients with more than about 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is about 3.0x l04cells / kg body weight for patients with less than or equal to 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is about 1.5 x 106cells for patients with more than 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is about 1x104cells / kg body weight, 2x 104cells / kg body weight, 3 x 104cells / kg body weight, 4 x 104cells / kg body weight, 5 x104cells / kg body weight, 6 x104cells / kg body weight, 7 x104cells / kg body weight, 8 x104cells / kg body weight, 9 x104cells / kg body weight, 10 x1048ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION cells / kg body weight, 11 x 104cells / kg body weight, 12 x 104cells / kg body weight, 13 x 104cells / kg body weight, 14 x 104cells / kg body weight, 15 x 104cells / kg body weight, 16 x 104cells / kg body weight, 17 x 104cells / kg body weight, 18 x 104cells / kg body weight, 19 x 104cells / kg body weight, 20 x 104cells / kg body weight, 21 x 104cells / kg body weight, 22 x 104cells / kg body weight, 23 x 104cells / kg body weight, 24 x 104cells / kg body weight, 25 x 104cells / kg body weight, 26 x 104cells / kg body weight, 27 x 104cells / kg body weight, 28 x 104cells / kg body weight, 29 x 104cells / kg body weight, 30 x 104cells / kg body weight, 40 x 104cells / kg body weight, 50 x 104cells / kg body weight, 60 x 104cells / kg body weight, 70 x 104cells / kg body weight, 80 x 104cells / kg body weight, or about 90 x 104cells / kg body weight. In some embodiments of the method of treating a viral disorder, the therapeutically effective amount of cells is about 0.5 x 106cells, 1 x106cells, 1. 5 x 106cells, 2 x 106cells, 2.5 x106cells, 3 x106cells, 3.5 x 106cells, 4 x106cells, 4.5 x 106cells, 5 x 106cells, 5.5 x 106cells, 6 x 106cells, 6.5 x 106cells, 7 x 106cells, 7.5 x 106cells, 8 x 106cells, 8.5 x 106cells, 9 x 106cells, 9.5 x 106cells, 10 x 106cells, 10.5 x 106cells, 11 x 106cells, 11.5 x 106cells, 12 x 106cells, 12.5 x106cells, 13 x 106cells, 13.5 x106cells, 14 x106cells,14.5 xl 06cells, 15 x 106cells, 16 x 106cells, 17 x 106cells, 18 x 106cells, 19 x 106cells, 20 x 106cells, 2l x 106cells, 22 x 106cells, 23 x 106cells, 24 x 106cells, 25 x 106cells, 26 xl 06cells, 27 x 106cells, 28x106cells, 29 x 106cells, 30 x 106cells, 31 x 106cells, 32 x 106cells, 33 x 106cells, 34x106cells, 35 x 106cells, 36 x 106cells, 37 x 106cells, 38 x 106cells, 39 x 106cells, 40 x 106cells, 41 x 106cells,42x 106cells, 43 x 106cells, 44 x 106cells, or 45 x 106cells.

[0014] In some embodiments, the disclosure relates to a nucleic acid molecule encoding a chimeric antigen receptor (CAR) molecule that binds to cells expressing an EBV antigen, an SL9 antigen,a Nef antigen, and a SARS antigen. In some embodiments, the CAR molecule binds to cell expressing an EBV antigen and an HLA molecule comprising HLA-A*02:01, or a function variant thereof. In some embodiments, the CAR molecule binds to cell expressing an SL9 antigen and an HLA molecule comprising HLA-A*02 or a function variant thereof. In some embodiments, the CAR molecule binds to cell expressing Nef antigen and an HLA molecule comprising HLA-A*24 or a function variant thereof. In some embodiments, the CAR molecule binds to cell expressing a SARS antigen and an HLA molecule comprising HLA- B*08 or a function variant thereof.

[0015] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ 9ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 10ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50).

[0016] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 11ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100).

[0017] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), 12ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150).

[0018] In some embodiments, the nucleic acid molecule comprises a first nucleic acid sequence encoding a peptide comprising KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), 13ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200).

[0019] In some embodiment, the first nucleic acid sequence encodes a peptide comprising an amino acid sequence with at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of one of SEQ ID NOS: 1 through 200.

[0020] In some embodiments, the first nucleic acid sequence encodes a peptide comprising GTHDKCENPKX1X2X3X4X5X6X7X8QNX9NVVX10TNKELEDIYDESN (SEQ ID NO:202), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 202, wherein X1X2X3X4X5X6 X7X8 is one of SEQ ID NOS: 1 through 200 or a functional variant thereof comprising about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200, and X9 and X10 are any amino acid. In some embodiments, the nucleic acid molecule further comprises a second nucleic acid sequence encoding a peptide comprising KEETKEVLKKFKEKVNQFX11EHAFDIINKYGDKEIX12NMMX13MLLWRVX14FRSFRI DANNVELIKFNIRVLDWIMAEADNDLSYFISQ (SEQ ID NO:203), or a functional variant 14ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION thereof comprising from about 75% sequence identity to SEQ ID NO: 203, wherein X11, X12, X13, and X14are any amino acid. In some embodiments, the nucleic acid molecule comprises the first nucleic acid sequence and a third nucleic acid sequence encoding a single chain antibody or antibody fragment that binds to a cell expressing CD3, the first nucleic acid sequence and the third nucleic acid sequence together encode a chimeric peptide comprising a first domain and a second domain, the first domain comprising the peptide encoded by the first nucleic acid sequence, and the second domain comprising the single chain antibody or antibody fragment that binds to a cell expressing CD3. In some embodiments, the nucleic acid molecule comprising the first and third nucleic acid sequences further comprises the second nucleic acid sequence, and the first domain comprises the peptide encoded by the first nucleic acid sequence and the peptide encoded by the second nucleic acid sequence.

[0021] In some embodiments, the disclosure provides a nucleic acid molecule encoding a chimeric antigen receptor (CAR) molecule, wherein said CAR molecule comprises: a first CAR comprising a first antigen binding domain which binds to NYESO-1 Kras_G12V, PHOX2B, or Prame; a first transmembrane domain; a first co-stimulatory signaling domain; and / or a first primary signaling domain.

[0022] In some embodiments of a nucleic acid encoding a CAR molecule disclosed herein, the nucleotide sequence encoding the first primary signaling domain is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 100% different from the nucleotide sequence encoding the second primary signaling domain. In some embodiments, the nucleotide sequence encoding the first primary signaling domain differs by at least 1 nucleotide, 10 nucleotides, 20 nucleotides, 30 nucleotides, 40 nucleotides, 50 nucleotides, 60 nucleotides, 70 nucleotides, 80 nucleotides, 90 nucleotides, 100 nucleotides, 150 nucleotides, 200 nucleotides or all nucleotides from the nucleotide sequence encoding the second primary signaling domain.

[0023] In some embodiments, the disclosure relates to a pharmaceutical composition comprising a nucleic acid and a pharmaceutically acceptable carrier. In some embodiments, the nucleic acid is as described above. In some embodiments, the nucleic acid is as described above. In some embodiments, the disclosure relates to a pharmaceutical composition comprising a peptide as described herein and a pharmaceutically acceptable carrier. In some embodiments, the peptide is encoded by a nucleic acid as summarized above. In some embodiments, the peptide is encoded by a nucleic acid as summarized below. In some embodiments, the disclosure relates to a pharmaceutical composition comprising a cell herein and a pharmaceutically acceptable carrier. 15ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0024] In some embodiments, the disclosure relates to a cell comprising a nucleic acid herein. In some embodiments, the nucleic acid is as summarized above. In some embodiments, the nucleic acid is as summarized below.

[0025] In some embodiments, the cell is an immune effector cell, e.g., T cell (e.g., CD3+, CD4+ or CD8+ T cell), or an NK cell. In some embodiments, the cell is a human cell.

[0026] In some embodiments, provided herein is a method of providing anti-tumor immunity, comprising administering to a subject in need thereof, an effective amount of a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, a CAR molecule disclosed herein, e.g., a dual CAR molecule disclosed herein.

[0027] In some emobidments, the disclosure relates to a pharmaceutical composition comprising a nucleic acid.

[0028] In some embodiments, the disclosure provides a method of treating a subject having a disease associated with an antigen, comprising administering to the subject in need thereof, an effective amount of a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, a CAR molecule disclosed herein, e.g., a dual CAR molecule.

[0029] In some embodiments, the disclosure provides a bispecific antigen binding domain, comprising a first antigen binding domain which binds to EBV, SL9, Nef, or SARS and a second antigen binding domain which binds to a co-stimulatory molecule. In some embodiments, the disclosure provides a cell comprising a CAR comprising a first domain and a second domain. In some embodiments, the first domain comprises one or more of SEQ ID NOS:1 through 200, or functional variants thereof. In some embodiments, the first domain comprises a SEQ ID NO:202, or functional variants thereof.

[0030] In some embodiments, the first domain comprises a SEQ ID NO: 203, or a functional variant thereof.

[0031] In some embodiments, the first domain comprises a combination of two or more of: (1) one of SEQ ID NOS: 1 through 200, or functional variants thereof; (2) SEQ ID NO: 202 or functional variant thereof; and SEQ ID NO: 203, or functional variant thereof.

[0032] In some embodiments, provided herein is a chimeric antigen receptor (CAR), comprising a monospecific or bispecific antigen binding domain described herein.

[0033] In yet another embodiment, the disclosure provides a nucleic acid encoding a chimeric antigen receptor (CAR), which comprises a monspecific or bispecific antigen binding domain described herein.

[0034] In some embodiments of the bispecific antigen binding domain described herein, e.g., a CAR comprising the bispecific antigen binding domain, or nucleic acid encoding a CAR 16ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION comprising the bispecific antigen binding domain, the first antigen binding domain can be upstream (e.g., in an NH2-terminal orientation) of the second antigen binding domain, or the first antigen binding domain can be downstream (e.g., in a COOH-terminal orientation) of the second antigen binding domain.

[0035] In some embodiments, each of the first antigen binding domain and second antigen binding domains comprise a scFv, e.g., a light chain variable (VL) domain and a heavy chain variable (VH) domain. In some embodiments, the first antigen binding domain comprises an scFv comprising a first VH (VHl) and a first VL (VLl ). In some embodiments, the second antigen binding domain comprises an scFv comprising a second VH (VH2) and a second VL (VL2).

[0036] In some embodiments, a bispecific antigen binding domain has any one of the following N terminal to C terminal configurations: VL1-VH1-VH2-VL2; VH1-VL1-VH2- VL2; VL1-VH1- VL2-VH2; VH1-VL1-VL2-VH2, VH2-VL2-VL1-VH1; VL2-VH2-VL1- VH1; VH2-VL2-VH1-VLl; or VL2-VH2-VH1-VL1.

[0037] In some embodiments, a CAR comprising a bispecific antigen binding domain comprises the amino acid sequence of SEQ ID NO: 202.

[0038] In some embodiments, the disclosure provides a vector comprising a nucleic acid sequence encoding a CAR molecule disclosed herein, a nucleic acid encoding a bispecific antigen binding domain disclosed herein, or a nucleic acid encoding a CAR comprising a bispecific antigen binding domain disclosed herein.

[0039] In some embodiments, provided herein is a pharmaceutical composition comprising a nucleic acid encoding a CAR molecule disclosed herein or a pharmaceutical composition comprising: CAR molecule disclosed herein. In some embodiments, the pharmaceutical composition comprises an pharmaceutically acceptable carrier, such as excipient, a carrier, a diluent and / or a stabilizer.

[0040] In yet another embodiment, the disclosure provides a pharmaceutical composition comprising a bispecific antigen binding domain disclosed herein, a CAR comprising a bispecific antigen binding domain disclosed herein, or a CAR nucleic acid encoding a bispecific antigen binding domain disclosed herein. In some embodiments, the pharmaceutical composition comprises an excipient, a carrier, a diluent and / or a stabilizer.

[0041] In some embodiments, provided herein is a method of providing anti-tumor immunity, comprising administering to a subject in need thereof, a therapeutically effective amount of a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, a CAR disclosed herein, e.g., a tandem CAR disclosed herein. 17ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0042] In some embodiments, the disclosure provides a method of treating a subject having a disease associated with an antigen, comprising administering to the subject in need thereof, an effective amount of a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, a CAR disclosed herein, e.g., a tandem CAR disclosed herein.

[0043] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following enumerated embodiments.

[0044] The disclosure also provides a nucleic acid molecule encoding a chimeric antigen receptor (CAR) molecule, wherein said CAR molecule comprises: (a) a first CAR comprising a first antigen binding domain which binds to an EBV antigen, an SL9 antigen,a Nef antigen, or a SARS antigen; a first transmembrane domain; a first co-stimulatory signaling domain; and / or a first primary signaling domain. In some embodiments, the CAR molecule further comprises: (b) a second CAR comprising a second antigen binding domain which binds to a second antigen; a second transmembrane domain; a second co- stimulatory domain; and / or a second primary signaling domain.

[0045] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first transmembrane domain is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 100% different from the nucleotide sequence encoding the second transmembrane domain.

[0046] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first transmembrane domain differs by at least 1 nucleotide, 10 nucleotides, 20 nucleotides, 30 nucleotides, 40 nucleotides, 50 nucleotides, 60 nucleotides, 70 nucleotides, 80 nucleotides, 90 nucleotides, 100 nucleotides, 150 nucleotides, 200 nucleotides or all nucleotides from the nucleotide sequence encoding the second transmembrane domain.

[0047] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first co-stimulatory signaling domain differs by at least 1 nucleotide, 10 nucleotides, 20 nucleotides, 30 nucleotides, 40 nucleotides, 50 nucleotides, 60 nucleotides, 70 nucleotides, 80 nucleotides, 90 nucleotides, 100 nucleotides, 120 nucleotides, or all nucleotides from the nucleotide sequence encoding the second co-stimulatory signaling domain. 18ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0048] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first primary signaling domain is at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 100% different from the nucleotide sequence encoding the second primary signaling domain.

[0049] Some embodiments include nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first primary signaling domain differs by at least 1 nucleotide, 10 nucleotides, 20 nucleotides, 30 nucleotides, 40 nucleotides, 50 nucleotides, 60 nucleotides, 70 nucleotides, 80 nucleotides, 90 nucleotides, 100 nucleotides, 150 nucleotides, 200 nucleotides, 250 nucleotides, 300 nucleotides or all nucleotides from the nucleotide sequence encoding the second primary signaling domain.

[0050] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the first CAR and / or the second CAR comprises a signal peptide, e.g., a peptide comprising a stretch of hydrophobic amino acids, e.g., 5-16 residues.

[0051] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the signal peptide is chosen from a CD8alpha signal peptide, an interleukin 2 signal peptide, a human albumin signal peptide, a human chymotrypsinogen signal peptide, a human trypsinogen-2 signal peptide or other similar signal peptides disclosed in Stern B. et al. “Improving mammalian cell factories : The selection of signal peptide has a major impact on recombinant protein synthesis and secretion in mammalian cells.” (2007).

[0052] The nucleic acid molecule of any of the preceding embodiments, wherein the protease cleavage site or internal ribosomal entry site is situated between the first CAR and the second CAR.

[0053] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the protease cleavage site is situated such that a cell can express a fusion protein comprising a first CAR and a second CAR, optionally wherein the fusion protein is processed into two peptides by proteolytic cleavage.

[0054] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first CAR and the nucleotide sequence encoding the second CAR are disposed on a single nucleic acid construct.

[0055] Some embodiments include nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide encoding the first CAR and the nucleic acid encoding the second CAR are disposed on the same vector.

[0056] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first CAR and the nucleotide 19ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION sequence encoding the second CAR are disposed on different nucleic acid constructs, e.g., the nucleotide sequence encoding the first CAR is disposed on a first nucleic acid construct, and the nucleotide sequence encoding the second CAR is disposed on a second nucleic acid construct.

[0057] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the first CAR is disposed on a first vector.

[0058] Some embodiments comprise the nucleic acid molecule of any of the preceding embodiments, wherein the nucleotide sequence encoding the second CAR is disposed on a second vector.

[0059] In some embodiments, the nucleic acid molecule comprises a viral element, e.g., a viral packaging element.

[0060] Some embodiments comprise a vector comprising the nucleic acid molecule of any of embodiments described herein.

[0061] Some embodiments comprise the vector of any of the preceding embodiments, wherein the vector is chosen from a DNA, a RNA, a plasmid, a lentivirus vector, adenoviral vector, or a retrovirus vector.

[0062] Some embodiments comprise a cell (e.g., an immune effector cell) comprising the vector of an embodiment herein, or the nucleic acid molecule of an embodiment herein.

[0063] Some embodiments comprise the cell of any of the preceding embodiments, wherein the cell comprises a nucleic acid encoding the CAR molecule. The cell of any of the preceding embodiments, comprising the nucleic acid molecule of any of the preceding embodiments.

[0064] Some embodiments comprise the cell of embodiment of any any of the preceding embodiments, wherein the cell is a human cell.

[0065] Some embodiments comprise a method of making a cell (e.g., an immune effector cell) comprising transducing an immune effector cell, e.g., a T cell or NK cell, with a vector of an embodiment herein.

[0066] Some embodiments comprise a method of making a cell (e.g., an immune effector cell) comprising introducing a nucleic acid molecule of any one of the preceding embodiments, into an immune effector cell, e.g., a T cell or NK cell.

[0067] Some embodiments comprise a method of generating a population of RNA- engineered cells comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, where the RNA comprises a nucleic acid molecule of any of the preceding embodiments. 20ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION The bispecific antigen bind S, wherein the first antigen binding domain can be upstream (e.g., in an N-terminal orientation) of the second antigen binding domain, or the first antigen binding domain can be downstream (e.g., in a C-terminal orientation) of the second antigen binding domain.

[0068] Some embodiments comprise the bispecific antigen binding domain of any of the preceding embodiments, wherein each of the first antigen binding domain and second antigen binding domains comprise a scFv, e.g., a light chain variable (VL) domain and a heavy chain variable (VH) domain. The bispecific antigen binding domain of any of the preceding embodiments, wherein the VH can be upstream or downstream of the VL. The bispecific antigen binding domain of any of the preceding embodiments, wherein the first antigen binding domain comprises an scFv comprising a first VH (VHl) and a first VL (VL1).

[0069] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments, wherein the first antigen binding domain comprises an scFv comprising a first VH (VHl) and a first VL (VL1), wherein the second antigen binding domain comprises an scFv comprising a second VH (VH2) and a second VL(VL2).

[0070] Some embodiments comprise the bispecific antigen binding domain of any any preceding embodiments, wherein the first antigen binding domain is arranged with VHl upstream of VLl.

[0071] Some embodiments comprise the bispecific antigen binding domain of any, wherein first antigen binding domain is arranged with VLl upstream of VHl.

[0072] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments, wherein the second antigen binding domain is arranged with VH2 upstream of VL2. The bispecific antigen binding domain of any preceding embodiments, wherein the second antigen binding domain is arranged with VL2 upstream of VH2.

[0073] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments wherein the antigen binding domain has the following N terminal to C terminal configuration: VL1-VH1-VH2-VL2. The bispecific antigen binding domain of any preceding embodiments wherein the antigen binding domain has the following N terminal to C terminal configuration: VH1-VL1-VH2-VL2.

[0074] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments, wherein the antigen binding domain has the following N terminal to C terminal configuration: VL1-VH1-VL2-VH2. 21ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0075] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments, wherein the antigen binding domain has the following N terminal to C terminal configuration:VH1-VL1-VL2-VH2.

[0076] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments, wherein a linker is disposed between the first antigen binding domain and the second antigen binding domain.

[0077] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments, wherein the linker is disposed between the scFv of the first antigen binding domain and the scFv of the second antigen binding domain.

[0078] Some embodiments comprise the bispecific antigen binding domain of any preceding embodiments, wherein the linker is disposed between: VHl and VH2 if the construct has the configuration of: VL1-VH1-VH2-VL2; VLl and VH2 if the construct has the configuration of: VH1-VL1-VH2-VL2; VHl and VL2 if the construct has the configuration ofVLl-VHl-VL2-VH2; or VLl and VL2 if the construct has the configuration of VH1-VL1- VL2-VH2.

[0079] In some embodiments, the bispecific antigen binding domain of any one of any preceding embodiments comprises a linker, wherein the linker is long enough to avoid mispairing between the domains of the two scFvs. In some embodiments, the linker is a (Gly4- Ser)n linker (SEQ ID NO: 251), wherein n is 1, 2, 3, 4, 5, or 6. In some embodiments, the bispecific antigen binding domain comprises a linker, wherein the linker comprises of the amino acid sequence: LAEAAAK (SEQ ID NO: 240).

[0080] In some embodiments, the disclosure relates to the bispecific antigen binding domain of any preceding embodiments, wherein a linker is disposed between the VL and VH of the scFv of the first antigen binding domain, e.g., a linker described herein.

[0081] In some embodiments, the disclosure relates to the bispecific antigen binding domain of any preceding embodiments, wherein a linker is disposed between the VL and VH of the scFv of the second antigen binding domain, e.g., a linker described herein.

[0082] In some embodiments, the disclosure relates to a bispecific chimeric antigen receptor (CAR), comprising the bispecific antigen binding domain of any one of the previously disclosed embodiments. A nucleic acid construct encoding a bispecific chimeric antigen receptor (CAR), wherein the nucleic acid construct encodes the bispecific antigen binding domain of any one chimeric antigen receptor (CAR), comprising a bispecific antigen binding domain which comprises: a first antigen binding domain which binds to an EBV antigen, an SL9 antigen,a Nef antigen, or a SARS antigen and a second antigen binding domain which 22ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION binds to CD137 and / or CD3, wherein the CAR comprises a transmembrane domain, a co- stimulatory domain and / or a primary signaling domain.

[0083] In some embodiments, the disclosure relates to a nucleic acid molecule of any of the preceding embodiments, wherein the protease cleavage site is situated such that a cell can express a fusion protein comprising a first CAR and a second CAR, optionally wherein the fusion protein is processed into two peptides by proteolytic cleavage. A CAR comprising the bispecific antigen binding domain of any one of the preceding embodiments.

[0084] In some embodiments, the disclosure relates to a CAR of the previous embodiment, comprising: a bispecific antigen binding domain; a transmembrane domain; and a co- stimulatory signaling domain; a bispecific antigen binding domain; a transmembrane domain; and a primary signaling domain; or a bispecific antigen binding domain; a transmembrane domain; a co-stimulatory signaling domain; and a first primary signaling domain.

[0085] In some embodiments, the disclosure relates to a CAR of any one of the previously disclosed embodiments, wherein the CAR comprises a transmembrane domain, wherein the transmembrane domain is chosen from the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CDS, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154.

[0086] In some embodiments, the disclosure relates to a CAR of any one of the previously disclosed embodiments, wherein the bispecific antigen binding domain is connected to the transmembrane domain by a hinge region, e.g., a hinge region described herein.

[0087] In some embodiments, the disclosure relates to a CAR of any one of the previously disclosed embodiments, wherein the CAR comprises a co- stimulatory domain, wherein the co-stimulatory domain comprises a signaling domain of OX40, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CDl la / CD18), ICOS (CD278) or 4-lBB (CD137).

[0088] In some embodiments, the disclosure relates to a CAR of any one of the previously disclosed embodiments, wherein the co-stimulatory domain comprises a 4-lBB signaling domain. The CAR of any one of the previously disclosed embodiments, wherein the CAR comprises a primary signaling domain comprising a signaling domain of CD3 zeta.

[0089] In some embodiments, the disclosure relates to a CAR of any one of the previously disclosed embodiments, wherein the CAR comprises (1) one or more amino acid sequence selected from SEQ ID NO: 1 through 200 and (2) an amino acid sequence of SEQ ID NO: 202, and / or (3) and amino acid sequence of SEQ ID NO: 203, or an amino acid sequence comprising at least about 95%, 96%, 97%, 98%, or 99% sequence identity thereto. 23ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0090] In some embodiments, the disclosure relates to a method of making a cell (e.g., an immune effector cell) comprising: transducing an immune effector cell, e.g., a T cell or NK cell, with a vector of embodiment 143; or introducing a CAR nucleic acid molecule of any one of embodiments 129 to 142, into an immune effector cell, e.g., a T cell or NK cell.

[0091] In some embodiments, the disclosure relates to a pharmaceutical composition comprising the nucleic acid encoding the CAR molecule of any one of the previously disclosed embodiments or the bispecific antigen binding domain of any one of embodiments, the CAR of any one of above identified embodiments.

[0092] In some embodiments, the disclosure relates to a method of providing anti-tumor immunity, comprising administering to a subject in need thereof, a therapeutically effective amount of a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, the nucleic acid encoding a CAR molecule of any one of embodiments according to the invention , the bispecific antigen binding domain of any one of embodiments according to the invention, the CAR of any one of embodiment according to the invention or the CAR nucleic acid of any one of embodiments according to the invention.

[0093] In some embodiments, the disclosure relates to a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, the nucleic acid encoding a CAR molecule of any one of the preceding embodiments, the bispecific antigen binding domain of any one of the preceding embodiments of the CAR of any one of the preceding embodiments, or the CAR nucleic acid of for use in a method of providing anti-tumor immunity to a subject.

[0094] In some embodiments, the disclosure relates to a method or the use of the previously disclosed two embodiments, wherein the cell is a T cell or an NK cell. The method or the use of the previously disclosed three embodiments, wherein the cell is a human cell. The method of any one of the preceding four embodiments, wherein the cell is an autologous cell or an allogeneic cell.

[0095] In some embodiments, the disclosure relates to a method of treating a subject having a disease associated with an antigen, comprising administering to the subject in need thereof, an effective amount of a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, a nucleic acid encoding a CAR molecule of any one of the disclosed embodiments, a bispecific antigen binding domain of any one of the disclosed embodiments, the CAR of any one of the disclosed embodiments or the CAR nucleic acid of any one of the disclosed embodiments.

[0096] In some embodiments, the disclosure relates to a kit comprising the cell as summarized above or a pharmaceutical composition as summarized above. 24ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0097] In some embodiments, the disclosure relates to a kit comprising a nucleic acid as disclosed herein and a cell capable of being transformed by the nucleic acid. BRIEF DESCRIPTION OF THE DRAWINGS

[0098] FIGS. 1A through 1D depict a determination of the specificity of [ TRACeR] _(MHC-I,A02)1'(NY-ESO-l) with different peptides. FIG. 1A. Alanine scan of the NY- ESO-1 antigen showing the requirement of S1, M4 and W5. Residues colored in yellow are solvent exposed and residues L2 L3, L6, T7, and V9 have side chains buried in MHC-1 groove. FIG. 1A discloses SEQ ID NOS 252, 252, and 262, respectively, in order of appearance. FIG. 1B. X-scan on the three key residues for TRACeR (left) and 1G4 TCR (right), MFI (mean fluorescence intensity) signal was normalized as a percentage of wild type binding signal. SI V peptide synthesis failed due to the hydrophobicity. FIG.1C. Correlation plot of the binding levels between TRACeR construct with a panel of 97 individual HLA-I allotypes upon incubation with the wild-type NY-ESO-1 or the non-binder NY-ESO-1WSA peptides. The dashed line represents a conceptual I:1 correlation (no difference between the peptides). FIG. 1D. Alignment ofMHC-1 and MRI structures. NY-ESO-1 peptide is shown in light grey (first position shown as stick) and the 5-OP-RU antigen is shown in grey. a schematic of the tandem constructs of the present disclosure. Each tandem CAR comprises a bispecific antigen binding domain comprising a NYESO-1 antigen binding domain and a CD3 antigen binding domain.

[0099] FIGS. 2A through 2F depict [TRACeR] _(MHC-I,A02)1'(NY-ESO-l)-antiCD3 scFv BiTE mediate target- specific cancer killing and CAR-T killing. FIG. 2A shows a Schematic representation of the BiTE construct.[(TRACeR)]_(MHC-I,A02)1'(NY-ESO- 1)recognizes NY-ESO-1 antigen presented by surface MHC-1, and the anti-CD3 scFv was used for the T cell binding module. FIG. 2B shows in vitro tumor cell killing assays with [TRACeR] _(MHC-I,A02)1'(NY-ESO- 1)-antiCD3 BiTEs or control BiTEs. Activated human total CD3+ cells were mixed 2:1 with tumor cell lines for 18 hours then analyzed for tumor specific killing by flow cytometry. Frequency oflive tumor cells were normalized to PBS treated control cells. One of two independent repeats is shown. FIGS.2C and 2D show anti- CD3 / anti-CD28 activated total T cells that were incubated with indicated tumor cell lines and treated with TRACeR BiTEs, control bispecific antibodies or PBS for 18 hours. Control bispecific antibodies were used at a concentration of 10 ng / mL. Cells were then isolated and analyzed by flow cytometry. Cells analyzed are live, CD3+CD4+ and CD3+CD8+ cells. FIG.2E. CD69 and 4-IBB expression in T cells incubated with HBL-1 25ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION cells FIG. 2 D: CD69 and 4-lBB expression in T cells incubated with HLY-1 cells. Statistical significance was determined by one-way ANOVA followed by Dunnett's multiple comparisons test. FIG. 2E is a schematic representation of the [TRACeR]_(MHC-I,A02Y'(NY-ESO-l )-CAR T cells recognizes NY-ESO-1 antigen presented by surface MHC-1. 2F. In vitro tumor cell killing assays with [TRACeR] _(MHC- I,A02)"'(NY-ESO-l)-CAR T cells or control IG4 TCR. Engineered T cells were mixed I: I with tumor cell lines for 18 hours then analyzed for tumor specific killing by flow cytometry. Frequency of live tumor cells co-cultured with engineered T cells were normalized to those co-cultured with untransduced T cells.

[0100] FIG. 3 depicts Circular permutation scheme for redesigning TRACeR scaffold in schematic and crystal forms, along with cell binding data with NY-ESO-1 peptide bound to activating cells. FIG.3 includes SEQ ID NOS 263 and 254, respectively, in order of appearance.

[0101] FIGS.4A through 4C depict Cross-allele specificity profiling of [TRACeR] _(MHC- I,A02)A(NY-ESO-l). FIG. 4A. Scatterplot of the BLOSUM62 sequence similarity to A*02:01 for HLA residues interfacing with the TRACeR versus the predicted percent rank binding affinity. The black dashed line represents a 5% rank binding affinity denoting a weak binder per NetMHCPan. FIG. 4B. Polymorphisms (red spheres) present in 10 HLAs (HLA-A*02:06, HLA- A*02:07, HLA-A*02:09, HLA- A*02:l l, HLA-A*02:131, HLA-A*02:l 7, HLA-A*02:18, HLA-A*02:240, HLA- A*02:448, HLA-A*02:642) relative to HLA-A*02:01 mapped onto the TRACeRNY- ESO-l ,A02 crystal structure. FIG. 4C. Binding levels of TRACeR construct with a panel of 97 individual HLA-1 allotypes upon incubation with the wild-type NY-ESO-1 or the non-binder NY-ESO-1 W5A peptides.

[0102] FIGS. 5A and 5B depict Structural alignment of predicted model and crystal structure of TRA CeRNr-Eso-1,Ao2. FIG.5A. Alignment of TRACeR helical bundle (yellow and green) with the hom MAM N terminal do(grey). FIG. 5B. Alignment of TRACeR monomeric binding mode with computational model (model is shown in shadow. FIG. 6 depicts SSM analysis of [TRACeR] _(MHC-I,A02)A(NY-ESO-l), non-circular permuted. Site saturate mutagenesis library of [TRACeR] _(MHC- I,A02)A(NY-ESO-l) was stained with 10 nM pMHC monomer and top 0.5% binding population was collected as child pool. Enrichment ratio of each mutation was calculated as (sequence count percentage in child pool) / (sequence count percentage in mother pool), normalized based on the enrichment ratio of wild type. Deep sequencing was 26ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION performed with Illumina 2*300 Kit at Stanford PAN facility. FIG.6 includes SEQ ID NO: 264.

[0104] FIG. 7 depicts a comparison of TRACeRZ, i!PAo1 to TCR:pHLA-I structures in the Protein Data Bank. TCR:nonamer / HLA-1 structural dataset was generated using amodified version ofHLA3DB1as described previously. A selection resulted in 67 crystalstructures. Complexes were analyzed using PDBePISA2as implemented in CCP4 (v.8.0)3to obtain peptide / receptor and HLA / receptor interface area values. Scatter plot depicting the interface area between the immune receptor and the HLA (x-axis) or the peptide (y-axis) (black: TCRs, grey: scFvs). The corresponding interface areas of the TRACeR with HLA- A*02:01 and NY-ESO-1 peptide is shown as an triangle.

[0105] FIG. 8 depicts purification of a〖TRACeR〗_(MHC-I,A02)^(NY-ESO-1)- antiCD3 BiTEs construct by the gel image and the dimer construct shows cancer killing.

[0106] FIGS. 9A through 9C depict designing dimerized TRACeR into monomer binder. FIG. 9A. Inspired by crystal structure, we redesigned the domain-swapped dimer into a monomer by connecting two monomers and removing one ARE site. Surface residues were redesigned with ProteinMPNN. FIG. 9B. Monomeric TRACeR purification SDS-PAGE gel and SEC curve (superdex 75). Monomeric TRACeR can be easily purified from E.coli and is highly soluble. FIG.9B. Monomeric TRACeR is still peptide specific based on yeast surface display (staining concentration: 50 nM tetramer) FIG.9C. Titration of monomeric [TRACeR] _(MHC-I,A02)A(NY-ESO-l) on T2 cells pulsing NY-ESO-1 peptide or EBV peptide as negative control. FIG. 9C. The connection scheme to create the rewired monomer. The equivalence to the crystal structure is denoted by the inverted labels on the helices (Hl, H2 and H3).

[0107] FIGS. l0A through l0C depict receptor design, expression and tetramer bindingof engineered T cell receptors. FIG. 10A. F I G . 10 A i n c l u d e s S E Q I D N O :265. Engineered CAR and TCR receptor construct design. FIG. 10B. Engineered receptorexpression 13 days post sorting. Quantification of median fluorescent intensity of anti-myc signal. FIG. 10C. Binding of engineered T cells to cognate HLA-A *02:01 human NY- ESO-1157-165 Cl 65V SLLMWITQV (SEQ ID NO: 252) or control HLA-A*02:0l EBV LMP2 426-434 CLGGLLTMV (SEQ ID NO: 254) tetramers. Quantification of median fluorescent intensity of each tetramer signal.

[0108] FIGS.11A through 11E depict engineered monomeric TRACeR-I shows multi- HLA allelic compatibility against viral targets FIG. 11A shows a rewiring scheme to 27ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION connect domain-swapped TRACeR into a monomer. FIG. 11B illustrates a model of rewired TRACeR on MHC-I. FIG. 11C shows SPR affinity measurements of the rewired tumor-targeting〖^^^^^^^^^^^^〗_(^^^^^^−^^, ^^02)^(^^^^−^^^^^^−1). FIG. 11D is an open-book view of the TRACeR interface with library positions colored in salmon and peptide in cyan. FIG. 11E illustrates TRACeR-I binding to a panel of diverse disease-relevant epitopic peptides presented by diverse HLAs. Final enrichment round before NGS is shown; multiple sequences are present in the pool. Staining concentration: on target, 25 nM tetramer (100 nM MHC + avidity); off target, 100 nM tetramer (400 nM MHC + avidity).

[0109] FIG. 12 depicts FACS enrichment of A02 / SL9-TRACeR mutagenesis library. HLA-A*02:01 / SL9 PE tetramers were used to stain for on-target binding.

[0110] FIG. 13 depicts binding kinetics of TRACeRs determined by Surface Plasmon Resonance (SPR). A concentration series of soluble TRACeRs binding (response units, RU) to immobilized HLA-A*02:01 / SL9 target over time (seconds). A global model (black lines) was fit to the binding data (colored lines) to determine kinetic parameters. Affinity maturation of TRACeR (A02 SL9) improved binding strength by several orders of magnitude from a KD of 46.3 μM to 60.8 nM. DETAILED DESCRIPTION Definitions

[0111] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

[0112] The term “a” and “an” refers to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0113] The term “about” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20%, ±10%, ±5%,, ±1%, or ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. For recitation of numeric ranges herein, each intervening number therebetween with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 28ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6,9, and 7.0 are explicitly contemplated.

[0114] As used herein, the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of subjects without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in theart. For example, Berge et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19.

[0115] The term “Chimeric Antigen Receptor,” a “CAR,” or a “CAR molecule” refers to a set of polypeptides, typically two in the simplest embodiments, which when in an immune effector cell, provides the cell with specificity for a target cell and with intracellular signal generation. In some embodiments, a CAR comprises at least an extracellular antigen binding domain, a transmembrane domain and a cytoplasmic signaling domain (also referred to herein as “an intracellular signaling domain”) comprising a functional signaling domain derived from a stimulatory molecule and / or costimulatory molecule as defined below. In some embodiments, the set of polypeptides are contiguous with each other, e.g., are in the same polypeptide chain, e.g., comprise a chimeric fusion protein. In some embodiments, the set of polypeptides are not contiguous with each other, e.g., are in different polypeptide chains. In some embodiments, the set of polypeptides include a dimerization switch that, upon the presence of a dimerization molecule, can couple the polypeptides to one another, e.g., can couple an antigen binding domain to an intracellular signaling domain.

[0116] In some embodiments, the stimulatory molecule is the zeta chain associated with the T cell receptor complex.

[0117] In some embodiments, the cytoplasmic signaling domain further comprises one or more functional signaling domains derived from at least one costimulatory molecule as defined below.

[0118] In some embodiments, the costimulatory molecule is chosen from the costimulatory molecules described herein, e.g., 4-IBB (i.e., CD137), CD27 and / or CD28. In some embodiments, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain derived from a stimulatory molecule.

[0119] In some embodiments, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain derived from a costimulatory molecule and 29ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION a functional signaling domain derived from a stimulatory molecule. In some embodiments, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising two functional signaling domains derived from one or more costimulatory molecule(s) and a functional signaling domain derived from a stimulatory molecule. In some embodiments, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising at least two functional signaling domains derived from one or more costimulatory molecule(s) and a functional signaling domain derived from a stimulatory molecule.

[0120] In some embodiment, the CAR comprises an optional leader sequence at the amino- terminus (N-ter) of the CAR fusion protein. In some embodiments, the CAR further comprises a leader sequence at the N-terminus of the extracellular antigen binding domain, wherein the leader sequence is optionally cleaved from the antigen binding domain (e.g., a scFv) during cellular processing and localization of the CAR to the cellular membrane.

[0121] The term “signaling domain” refers to the functional portion of a protein which acts by transmitting information within the cell to regulate cellular activity via defined signaling pathways by generating second messengers or functioning as effectors by responding to such messengers.

[0122] As used herein, the term “binding domain” refers to a protein, e.g., an immunoglobulin chain or fragment thereof, comprising at least one immunoglobulin variable domain sequence. The term “binding domain” (also referred to herein as “antibody molecule”) encompasses antibodies and antibody fragments. In some embodiments an antibody molecule is a multispecific antibody molecule, e.g., it comprises a plurality of immunoglobulin variable domain sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In some embodiments, a multispecific antibody molecule is a bispecific antibody molecule. A bispecific antibody has specificity for no more than two antigens. A bispecific antibody molecule is characterized by a first immunoglobulin variable domain sequence which has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope.

[0123] The term “antibody fragment” refers to at least one portion of an antibody, that retains the ability to specifically interact with (e.g., by binding, steric hinderance, stabilizing / destabilizing, spatial distribution) an epitope of an antigen. Examples of antibody 30ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION fragments include, but are not limited to, Fab, Fab’, F(ab’)2, Fv fragments, scFv antibody fragments, disulfide-linked Fvs (sdFv), a Fd fragment consisting of the VH and CHI domains, linear antibodies, single domain antibodies such as sdAb (either VL or VH), camelid VHH domains, multi-specific antibodies formed from antibody fragments such as a bivalent fragment comprising two Fab fragments linked by a disulfide brudge at the hinge region, and an isolated CDR or other epitope binding fragments of an antibody. An antigen binding fragment can also be incorporated into single domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv (see, e.g., Hollinger and Hudson, Nature Biotechnology 23:1126-1136, 2005). Antigen binding fragments can also be grafted into scaffolds based on polypeptides such as a fibronectin type III (Fn3) (see U.S. Patent No.: 6,703,199, which describes fibronectin polypeptide minibodies). The term “scFv” refers to a fusion protein comprising at least one antibody fragment comprising a variable region of a light chain and at least one antibody fragment comprising a variable region of a heavy chain, wherein the light and heavy chain variable regions are contiguously linked, e.g., via a synthetic linker, e.g., a short flexible polypeptide linker, and capable of being expressed as a single chain polypeptide, and wherein the scFv retains the specificity of the intact antibody from which it is derived. Unless specified, as used herein an scFv may have the VL and VH variable regions in either order, e.g., with respect to the N- terminal and C-terminal ends of the polypeptide, the scFv may comprise VL-linker-VH or may comprise VH-linker-VL.

[0124] The term “complementarity determining region” or “CDR,” as used herein, refers to the sequences of amino acids within antibody variable regions which confer antigen specificity and binding affinity. The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of lmmunological Interest,” 5thEd. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering scheme), Al-Lazikani et al., (1997) JMB 273, 927-948 (“Chothia” numbering scheme) and ImMunoGenTics (IMGT) numbering (Lefranc, M.-P., The Immunologist, 7, 132-136 (1999); Lefranc, M.-P. et al., Dev. Comp. Immunol., 27, 55-77 (2003) (“IMGT” numbering scheme). For example, for classic formats, under Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDRl ), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDRl), 50-56 (LCDR2), and 89-97 (LCDR3). Under Chothia, the CDR amino acids in the VH are numbered 26-32 (HCDRl ), 52- 56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in VL 31ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION are numbered 26-32 (LCDRl ), 50-52 (LCDR2), and 91-96 (LCDR3). By combining the CDR definitions of both Kabat and Chothia, the CDRs consist of amino acid residues 26-35 (HCDRl ), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDRl ), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL. Under IMGT, the CDR amino acid residues in the VH are numbered approximately 26-35 (CDRl), 51-57 (CDR2) and 93-102 (CDR3), and the CDR amino acid residues in the VL are numbered approximately 27-32 (CDRl), 50-52 (CDR2), and 89-97 (CDR3) (numbering according to “IMGT”). Under IMGT, the CDR regions of an antibody can be determined using the program IMGT / DomainGap Align.

[0125] The portion of the CAR of the invention comprising an antibody or antibody fragment thereof may exist in a variety of forms where the antigen binding domain is expressed as part of a contiguous polypeptide chain including, for example, a single domain antibody fragment (sdAb), a single chain antibody (scFv), a humanized antibody, or bispecific antibody (Harlow et al., 1999, In: Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, NY; Harlow et al., 1989, In: Antibodies: A Laboratory Manual, Cold Spring Harbor, New York; Houston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883; Bird et al., 1988, Science 242:423-426). In some embodiments, the antigen binding domain of a CAR composition of the invention comprises an antibody fragment. In a further embodiment, the CAR comprises an antibody fragment that comprises a scFv.

[0126] The term “antibody heavy chain,” refers to the larger of the two types of polypeptide chains present in antibody molecules in their naturally occurring conformations, and which normally determines the class to which the antibody belongs.

[0127] The term “antibody light chain,” refers to the smaller of the two types of polypeptide chains present in antibody molecules in their naturally occurring conformations. Kappa (x) and lambda (1-) light chains refer to the two major antibody light chain isotypes.

[0128] The term “recombinant antibody” refers to an antibody which is generated using recombinant DNA technology, such as, for example, an antibody expressed by a bacteriophage or yeast expression system. The term should also be construed to mean an antibody which has been generated by the synthesis of a DNA molecule encoding the antibody and which DNA molecule expresses an antibody protein, or an amino acid sequence specifying the antibody, wherein the DNA or amino acid sequence has been obtained using recombinant DNA or amino acid sequence technology which is available and well known in the art.

[0129] The term “antigen” or “Ag” refers to a molecule that provokes an immune response. This immune response may involve either antibody production, or the activation of specific 32ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION immunologically-competent cells, or both. The skilled artisan will understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA. A skilled artisan will understand that any DNA, which comprises a nucleic acid sequence or a partial nucleic acid sequence encoding a protein that elicits an immune response therefore encodes an “antigen” as that term is used herein. Furthermore, one skilled in the art will understand that an antigen need not be encoded solely by a full length nucleic acid sequence of a gene. It is readily apparent that the present disclosure includes, but is not limited to, the use of partial nucleic acid sequences of more than one gene and that these nucleic acid sequences are arranged in various combinations to encode polypeptides that elicit the desired immune response. Moreover, a skilled artisan will understand that an antigen need not be encoded by a “gene” at all. It is readily apparent that an antigen can be generated, synthesized, or can be derived from a biological sample, or might be macromolecule besides a polypeptide. Such a biological sample can include, but is not limited to a tissue sample, a tumor sample, a cell or a fluid with other biological components.

[0130] The term “anti-viral effect” refers to a biological effect which can be manifested by various means, including but not limited to, e.g., a decrease in number of virus infected cells, an increase in life expectancy, decrease in viral infected cell proliferation, or amelioration of various physiological symptoms associated with the viral disorder condition. An “anti-viral effect” can also be manifested by the ability of the peptides, polynucleotides, cells and antibodies of the invention in prevention of the occurrence of a viral disorder in the first place.

[0131] The term “allogeneic” refers to any material derived from a different animal of the same species as the individual to whom the material is introduced. Two or more individuals are said to be allogeneic to one another when the genes at one or more loci are not identical. In some embodiments, allogeneic material from individuals of the same species may be sufficiently unlike genetically to interact antigenically.

[0132] The term “xenogeneic” refers to a graft derived from an animal of a different species.

[0133] The term “combination” refers to either a fixed combination in one dosage unit form, or a combined administration where a compound of the present disclosure and a combination partner (e.g. another drug as explained below, also referred to as “therapeutic agent” or “co-agent”) may be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative, e.g. synergistic effect. The single components may be packaged in a kit or 33ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION separately. One or both of the components (e.g., powders or liquids) may be reconstituted or diluted to a desired dose prior to administration. The terms “co-administration” or “combined administration” or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g. a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time. The term “pharmaceutical combination” as used herein means a product that results from the mixing or combining of more than one therapeutic agent and includes both fixed and non-fixed combinations of the therapeutic agents. The term “fixed combination” means that the therapeutic agents, e.g. a compound of the present disclosure and a combination partner, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non- fixed combination” means that the therapeutic agents, e.g. a compound of the present disclosure and a combination partner, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g. the administration of three or more therapeutic agent.

[0134] The term “cancer” refers to a disease characterized by the rapid and uncontrolled growth of aberrant cells. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. Examples of various cancers are described herein and include but are not limited to, breast cancer, prostate cancer, ovarian cancer, cervical cancer, skin cancer, pancreatic cancer, colorectal cancer, renal cancer, liver cancer, brain cancer, lymphoma, leukemia, lung cancer and the like. The terms “tumor” and “cancer” are used interchangeably herein, e.g., both terms encompass solid and liquid tumors. As used herein, the term “cancer” or “tumor” includes premalignant, as well as malignant cancers and tumors.

[0135] As used herein, unless otherwise specified, the terms “prevent,” “preventing” and “prevention” refer to an action that occurs before the subject begins to suffer from the condition, or relapse of the condition. Prevention need not result in a complete prevention of the condition; partial prevention or reduction of the condition or a symptom of the condition, or reduction of the risk of developing the condition, is encompassed by this term.

[0136] Administered “in combination”, as used herein, means that two (or more) different treatments are delivered to the subject during the course of the subject’s affliction with the disorder, e.g., the two or more treatments are delivered after the subject has been diagnosed with the disorder and before the disorder has been cured or eliminated or treatment has ceased 34ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION for other reasons. In some embodiments, the delivery of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as “simultaneous” or “concurrent delivery”. In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments of either case, the treatment is more effective because of combined administration. For example, the second treatment is more effective, e.g., an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in the absence of the first treatment, or the analogous situation is seen with the first treatment. In some embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive. The delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered. In some embodiments, the CAR-expressing cell is administered at a dose and / or dosing schedule described herein, and the B-cell inhibitor, or agent that enhances the activity of the CDI 9 CAR-expressing cell is administered at a dose and / or dosing schedule described herein.

[0137] “Derived from” as that term is used herein, indicates a relationship between a first and a second molecule. It generally refers to structural similarity between the first molecule and a second molecule and does not connote or include a process or source limitation on a first molecule that is derived from a second molecule. For example, in the case of an intracellular signaling domain that is derived from a CD3zeta molecule, the intracellular signaling domain retains sufficient CD3zeta structure such that is has the required function, namely, the ability to generate a signal under the appropriate conditions. It does not connote or include a limitation toa particular process of producing the intracellular signaling domain, e.g., it does not mean that, to provide the intracellular signaling domain, one must start with a CD3zeta sequence and delete unwanted sequence, or impose mutations, to arrive at the intracellular signaling domain.

[0138] The term “conservative sequence modifications” refers to amino acid modifications that do not significantly affect or alter the binding characteristics of the antibody or antibody fragment containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions and deletions. Modifications can be introduced into an antibody or antibody fragment of the invention by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue 35ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within a CAR of the invention can be replaced with other amino acid residues from the same side chain family and the altered CAR can be tested using the functional assays described herein.

[0139] The term “stimulation,” refers to a primary response induced by binding of a stimulatory molecule (e.g., a TCR / CD3 complex or CAR) with its cognate ligand (or tumor antigen in the case of a CAR) thereby mediating a signal transduction event, such as, but not limited to, signal transduction via the TCR / CD3 complex or signal transduction via the appropriate NK receptor or signaling domains of the CAR. Stimulation can mediate altered expression of certain molecules.

[0140] The term “stimulatory molecule,” refers to a molecule expressed by an immune cell, e.g., T cell, NK cell, or B cell, that provides the cytoplasmic signaling sequence(s) that regulates activation of the immune cell in a stimulatory way for at least some embodiment of the immune cell signaling pathway. In some embodiments, the signal is a primary signal that is initiated by, for instance, binding of a TCR / CD3 complex with an MHC molecule loaded with peptide, and which leads to mediation of a T cell response, including, but not limited to, proliferation, activation, differentiation, and the like. A primary cytoplasmic signaling sequence (also referred to as a “primary signaling domain”) that acts in a stimulatory manner may contain a signaling motif which is known as immunoreceptor tyrosine-based activation motif or ITAM. Examples of an ITAM containing cytoplasmic signaling sequence that is of particular use in the invention includes, but is not limited to, those derived from CD3 zeta, common FcR gamma (FCERI G), Fc gamma Rlla, FcR beta (Fc Epsilon RI b), CD3 gamma, CD3 delta , CD3 epsilon, CD79a, CD79b, DAPI0, and DAP12. In a specific CAR of the invention, the intracellular signaling domain in any one or more CARS of the invention comprises an intracellular signaling sequence, e.g., a primary signaling sequence of CD3-zeta. In a specific CAR of the invention, the primary signaling sequence of CD3-zeta is the sequence, or the equivalent residues from a non-human species, e.g., mouse, rodent, monkey, ape and the like. 36ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0141] The term “antigen presenting cell” or “APC” refers to an immune system cell such as an accessory cell (e.g., a B-cell, a dendritic cell, and the like) that displays a foreign antigen complexed with major histocompatibility complexes (MHC’s) on its surface. T-cells may recognize these complexes using their T-cell receptors (TCRs). APCs process antigens and present them to T-cells.

[0142] “Immune effector cell,” as that term is used herein, refers to a cell that is involved in an immune response, e.g., in the promotion of an immune effector response. Examples of immune effector cells include T cells, e.g., alpha / beta T cells and gamma / delta T cells, B cells, natural killer (NK) cells, natural killer T (NK-T) cells, mast cells, and myeloid-derived phagocytes.

[0143] “Immune effector function or immune effector response,” as that term is used herein, refers to function or response, e.g., of an immune effector cell, that enhances or promotes an immune attack of a target cell. E.g., an immune effector function or response refers a property of a T or NK cell that promotes killing or the inhibition of growth or proliferation, of a target cell. In the case of a T cell, primary stimulation and co-stimulation are examples of immune effector function or response.

[0144] An “intracellular signaling domain,” as the term is used herein, refers to an intracellular portion of a molecule. The intracellular signaling domain generates a signal that promotes an immune effector function of the CAR containing cell, e.g., a CART cell or CAR- expressing NK cell. Examples of immune effector function, e.g., in a CART cell or CAR- expressing NK cell, include cytolytic activity and helper activity, including the secretion of cytokines.

[0145] In some embodiments, the intracellular signaling domain can comprise a primary intracellular signaling domain. Exemplary primary intracellular signaling domains include those derived from the molecules responsible for primary stimulation, or antigen dependent simulation. In some embodiments, the intracellular signaling domain can comprise a costimulatory intracellular domain. Exemplary costimulatory intracellular signaling domains include those derived from molecules responsible for costimulatory signals, or antigen independent stimulation. For example, in the case of a CART, a primary intracellular signaling domain can comprise a cytoplasmic sequence of a T cell receptor, and a costimulatory intracellular signaling domain can comprise cytoplasmic sequence from co-receptor or costimulatory molecule.

[0146] A primary intracellular signaling domain can comprise a signaling motif which is known as an immunoreceptor tyrosine-based activation motif or ITAM. Examples of ITAM- 37ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION containing primary cytoplasmic signaling sequences include, but are not limited to, those derived from CD3 zeta, common FcR gamma (FCERl G), Fc gamma Rlla, FcR beta (Fc Epsilon Rl b), CD3 gamma, CD3 delta, CD3 epsilon, CD79a, CD79b, DAPlO and DAP12.

[0147] The term “zeta” or alternatively “zeta chain”, “CD3-zeta” or “TCR-zeta” is defined as the protein provided as GenBan Acc. No. BAG36664. l, or the equivalent residues from a non-human species, e.g., mouse, rodent, monkey, ape and the like, and a “zeta stimulatory domain” or alternatively a “CD3-zeta stimulatory domain” or a “TCR-zeta stimulatory domain” is defined as the amino acid residues from the cytoplasmic domain of the zeta chain or functional derivative thereof, that are sufficient to functionally transmit an initial signal necessary for T cell activation. In one embodiment the cytoplasmic domain of zeta comprises residues 52 through 164 of GenBank Acc. No. BAG36664. l or the equivalent residues from a non-human species, e.g., mouse, rodent, monkey, ape and the like, that are functional orthologs thereof. In some embodiments, the “zeta stimulatory domain” or a “CD3-zeta stimulatory domain” is the sequence provided as SEQ ID NO: 96.

[0148] The term “costimulatory molecule” refers to the cognate binding partner on a T cell that specifically binds with a costimulatory ligand, thereby mediating a costimulatory response by the T cell, such as, but not limited to, proliferation. Costimulatory molecules are cell surface molecules other than antigen receptors or their ligands that contribute to an efficient immune response. Costimulatory molecules include, but are not limited to an MHC class I molecule, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, 64ignalling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-l(CDI la / CD18), 4-IBB (CD137), B7-H3, CDS, ICAM-1, ICOS 5 (CD278), GITR, BAFFR,LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRFI), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLAI, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CDI Id, ITGAE, CD103, ITGAL, CDI la, LFA-1, ITGAM, CDI lb, ITGAX, CDI le, ITGBI, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE / RANKL, DNAMI (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAMI, CRTAM, Ly9 (CD229), CD160 (BY55), PSGLI, CDI00 (SEMA4D), CD69, SLAMF6 (NTB-A, Lyl08), SLAM (SLAMFI, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG / Cbp, CD19a, and a ligand that specifically binds with CD83.

[0149] A costimulatory intracellular signaling domain can be the intracellular portion of a costimulatory molecule. A costimulatory molecule can be represented in the following protein 38ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION families: TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), and activating NK cell receptors. Examples of such molecules include CD27, CD28, 4-IBB (CD137), OX40, GITR, CD30, CD40, ICOS, BAFFR, HVEM, ICAM-1, lymphocyte function-associated antigen-I (LFA-1), CD2, 25CSD, CD7, CD287, LIGHT, NKG2C, NKG2D, SLAMF7, NKp80, NKp30, NKp44, NKp46, CD160, B7-H3, and a ligand that specifically binds with CD83, and the like.

[0150] The intracellular signaling domain can comprise the entire intracellular portion, or the entire native intracellular signaling domain, of the molecule from which it is derived, or a functional fragment or derivative thereof.

[0151] The term “4-lBB” refers to a member of the TNFR superfamily with an amino acid sequence provided as GenBank Acc. No. AAA62478.2, or the equivalent residues from a non- human species, e.g., mouse, rodent, monkey, ape and the like; and a “4-lBB costimulatory domain” is defined as amino acid residues 214-255 of GenBank Accession No. AAA62478.2, or the equivalent residues from a non-human species, e.g., mouse, rodent, monkey, ape and the like. In some embodiments, the “4-lBB costimulatory domain” is the sequence provided as SEQ ID NO:4 or the equivalent residues from a non-human species, e.g., mouse, rodent, monkey, ape and the like.

[0152] The term “encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (e.g., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene, cDNA, or RNA, encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleic acid sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

[0153] Unless otherwise specified, a “nucleic acid sequence encoding an amino acid sequence” includes all nucleic acid sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleic acid sequence that encodes a protein or a RNA may also include introns to the extent that the nucleic acid sequence encoding the protein may in some version contain an intron(s). 39ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0154] The term “effective amount” or “therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result.

[0155] The term “endogenous” refers to any material from or produced inside an organism, cell, tissue or system.

[0156] The term “exogenous” refers to any material introduced from or produced outside an organism, cell, tissue or system.

[0157] The term “expression” refers to the transcription and / or translation of a particular nucleic acid sequence driven by a promoter.

[0158] The term “transfer vector” refers to a composition of matter which comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell.

[0159] Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “transfer vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to further include non-plasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, a polylysine compound, liposome, and the like. Examples of viral transfer vectors include, but are not limited to, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like.

[0160] The term “expression vector” refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleic acid sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, including cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., lentiviruses, retroviruses, adenoviruses, and adeno- associated viruses) that incorporate the recombinant polynucleotide.

[0161] The term “lentivirus” refers to a genus of the Retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect non-dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient methods of a gene delivery vector. HIV, SIV, and FIV are all examples of lentiviruses.

[0162] The term “lentiviral vector” refers to a vector derived from at least a portion of a lentivirus genome, including especially a self-inactivating lentiviral vector as provided in 40ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION Milone et al., Mol. Ther. 17(8): 1453-1464 (2009). Other examples of lentivirus vectors that may be used in the clinic, include but are not limited to, e.g., the LENTIVECTOR® gene delivery technology from Oxford BioMedica, the LENTIMAX™ vector system from Lentigen and the like. Nonclinical types of lentiviral vectors are also available and would be known to one skilled in the art.

[0163] The “percent identity” or “percent homology” of two polynucleotide or two polypeptide sequences is determined by comparing the sequences using the GAP computer program (a part of the GCG Wisconsin Package, version 10.3 (Accelrys, San Diego, Calif.)) using its default parameters. “Identical” or “identity” as used herein in the context of two or more nucleic acids or amino acid sequences, may mean that the sequences have a specified percentage of residues that are the same over a specified region. The percentage may be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) may be considered equivalent. Identity may he performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0. Briefly, the BLAST algorithm, which stands for Basic Local Alignment Search Tool is suitable for determining sequence similarity. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov). This algorithm involves first identifying high scoring sequence pair (HSPs) by identifying short words of length Win the query sequence that either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Extension for the word hits in each direction are halted when: 1) the cumulative alignment score falls off by the quantity X from its maximum achieved value; 2) the cumulative score goes to zero or below, due to the accumulation of one or more negative- scoring residue alignments; or 3) the end of either sequence is reached. The Blast algorithm 41ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION parameters W, T and X determine the sensitivity and speed of the alignment. The Blast program uses as defaults a word length (W) of 11, the BLOSUM62 scoring matrix (see Henikoff et al., Proc. Natl. Acad. Sci. USA, 1992, 89, 10915-10919, which is incorporated herein by reference in its entirety) alignments (B) of 50, expectation (E) of 10, M=5, N=4, and a comparison of both strands. The BLAST algorithm (Karlin et al., Proc. Natl. Acad. Sci. USA, 1993, 90, 5873- 5787, which is incorporated herein by reference in its entirety) and Gapped BLAST perform a statistical analysis of the similarity between two sequences. One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide sequences or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to another if the smallest sum probability in comparison of the test nucleic acid to the other nucleic acid is less than about 1, less than about 0.1, less than about 0.01, and less than about 0.001. Two single-stranded polynucleotides are “the complement” of each other if their sequences can be aligned in an anti-parallel orientation such that every nucleotide in one polynucleotide is opposite its complementary nucleotide in the other polynucleotide, without the introduction of gaps, and without unpaired nucleotides at the 5’ or the 3’ end of either sequence. A polynucleotide is “complementary” to another polynucleotide if the two polynucleotides can hybridize to one another under moderately stringent conditions. Thus, a polynucleotide can be complementary to another polynucleotide without being its complement.

[0164] The term “homologous” or “identity” refers to the subunit sequence identity between two polymeric molecules, e.g., between two nucleic acid molecules, such as, two DNA molecules or two RNA molecules, or between two polypeptide molecules. When a subunit position in both of the two molecules is occupied by the same monomeric subunit; e.g., if a position in each of two DNA molecules is occupied by adenine, then they are homologous or identical at that position. The homology between two sequences is a direct function of the number of matching or homologous positions; e.g., if half (e.g., five positions in a polymer ten subunits in length) of the positions in two sequences are homologous, the two sequences are 50% homologous; if 90% of the positions (e.g., 9 of I 0), are matched or homologous, the two sequences are 90% homologous.

[0165] “Humanized” forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab’, F(ab’)2 or other antigen-binding subsequences of antibodies), which contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies and antibody fragments thereof are human immunoglobulins (recipient antibody or antibody fragment) in 42ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION which residues from a complementarity-determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity, and capacity. In some instances, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, a humanized antibody / antibody fragment can comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications can further refine and optimize antibody or antibody fragment performance. In general, the humanized antibody or antibody fragment thereof will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or a significant portion of the FR regions are those of a human immunoglobulin sequence. The humanized antibody or antibody fragment can also comprise at least a portion of an immunoglobulin constant region(Fe), typically that of a human immunoglobulin. For further details, see Jones et al., Nature, 321: 522-525, 1986; Reichmann et al., Nature, 332: 323-329, 1988; Presta, Curr. Op. Struct. Biol., 2:593-596, 1992.

[0166] “Fully human” refers to an immunoglobulin, such as an antibody or antibody fragment, where the whole molecule is of human origin or consists of an amino acid sequence identical to a human form of the antibody or immunoglobulin.

[0167] “Murine” refers to mice or rats. For example, a murine antibody or fragment thereof contains the sequence of an antibody or fragment thereof that is isolated from a murine animal, e.g., mouse or rat.

[0168] The term “isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.

[0169] In the context of the present disclosure, the following abbreviations for the commonly occurring nucleic acid bases are used. “A” refers to adenosine, “C” refers to cytosine, “G” refers to guanosine, “T” refers to thymidine, and “U” refers to uridine.

[0170] The term “operably linked” or “transcriptional control” refers to functional linkage between a regulatory sequence and a heterologous nucleic acid sequence resulting in expression of the latter. For example, a first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked 43ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Operably linked DNA sequences can be contiguous with each other and, e.g., where necessary to join two protein coding regions, are in the same reading frame.

[0171] The term “parenteral” administration of an immunogenic composition includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, intratumoral, or infusion techniques.

[0172] The term “nucleic acid” or “polynucleotide” refers to deoxyribonucleic acids (DNA) or ribonucleic acids (RNA) and polymers thereof in either single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementarity sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res.19:5081 (1991); Ohtsuka et al., J. Biol. Chem.260:2605-2608 (1985); and Rossolini et al., Mol. Cell. Probes 8:91-98 (1994)).

[0173] The terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein’s or peptide’s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. A polypeptide includes a natural peptide, a recombinant peptide, or a combination thereof.

[0174] The term “promoter” refers to a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence. 44ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0175] The term “promoter / regulatory sequence” refers to a nucleic acid sequence that is required for expression of a gene product operably linked to the promoter / regulatory sequence. In some instances, this sequence may be the core promoter sequence and in other instances, this sequence may also include an enhancer sequence and other regulatory elements which are required for expression of the gene product. The promoter / regulatory sequence may, for example, be one that expresses the gene product in a tissue specific manner.

[0176] The term “constitutive” promoter refers to a nucleic acid sequence that, when operably linked with a polynucleotide that encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell.

[0177] The term “inducible” promoter refers to a nucleic acid sequence that, when operably linked with a polynucleotide that encodes or specifies a gene product, causes the gene product to 10 be produced in a cell substantially only when an inducer that corresponds to the promoter is present in the cell.

[0178] The term “tissue-specific” promoter refers to a nucleic acid sequence that, when operably linked with a polynucleotide that encodes or specifies a gene product, causes the gene product to be produced in a cell substantially only if the cell is a cell of the tissue type corresponding to the promoter.

[0179] The term “flexible polypeptide linker” or “linker” as used in the context of a scFv refers to a peptide linker that consists of amino acids such as glycine and / or serine residues used alone or in combination, to link variable heavy and variable light chain regions together. In some embodiments, the flexible polypeptide linker is a Gly / Ser linker and comprises the amino acid sequence (Gly-Gly-Gly-Ser) (SEQ ID NO: 289), repeated n times where n is a positive integer equal to or greater than 1. For example, n=l, n=2, n=3. N=4, n=5 and n=6, n=7, n=8, n=9 and n=l0. In some embodiments, the flexible polypeptide linker is (Gly4 Ser)3 (SEQ ID NO: 282). In some embodiments, the linkers include multiple repeats of (Gly2Ser), and (GlySer). In some embodiments, the polypeptide does not include a linker, e.g., (n=0). Also included within the scope of the invention are linkers described in WO2012 / 138475, incorporated herein by reference).

[0180] As used herein, a 5’ cap (also termed an RNA cap, an RNA 7-methylguanosine cap or an RNA m7G cap) is a modified guanine nucleotide that has been added to the “front” or 5’ end of a eukaryotic messenger RNA shortly after the start of transcription. The 5’ cap consists of a terminal group which is linked to the first transcribed nucleotide. Its presence is critical for recognition by the ribosome and protection from RNAses. Cap addition is coupled to transcription, and occurs co-transcriptionally, such that each influences the other. Shortly after 45ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION the start of transcription, the 5’ end of the mRNA being synthesized is bound by a cap- synthesizing complex associated with RNA polymerase. This enzymatic complex catalyzes the chemical reactions that are required for mRNA capping. Synthesis proceeds as a multi-step biochemical reaction. The capping moiety can be modified to modulate functionality of mRNA such as its stability or efficiency of translation.

[0181] As used herein, “in vitro transcribed RNA” refers to RNA, preferably mRNA, that has been synthesized in vitro. Generally, the in vitro transcribed RNA is generated from an in vitro transcription vector. The in vitro transcription vector comprises a template that is used to generate the in vitro transcribed RNA

[0182] As used herein, a “poly(A)” is a series of adenosines attached by polyadenylation to the mRNA. In the preferred embodiment of a construct for transient expression, the polyA is between 50 and 5000 (SEQ ID NO: 255), preferably greater than 64, more preferably greater than 100, most preferably greater than 300 or 400. Poly(A) sequences can be modified chemically or enzymatically to modulate mRNA functionality such as localization, stability or efficiency of translation.

[0183] As used herein, “polyadenylation” refers to the covalent linkage of a polyadenylyl moiety, or its modified variant, to a messenger RNA molecule. In eukaryotic organisms, most messenger RNA (mRNA) molecules are polyadenylated at the 3’ end. The 3’ poly(A) tail is a long sequence of adenine nucleotides (often several hundred) added to the pre- mRNA through the action of an enzyme, polyadenylate polymerase. In higher eukaryotes, the poly(A) tail is added onto transcripts that contain a specific sequence, the polyadenylation signal. The poly(A) tail and the protein bound to it aid in protecting mRNA from degradation by exonucleases. Polyadenylation is also important for transcription termination, export of the mRNA from the nucleus, and translation. Polyadenylation occurs in the nucleus immediately after transcription of DNA into RNA, but additionally can also occur later in the cytoplasm. After transcription has been terminated, the mRNA chain is cleaved through the action of an endonuclease complex associated with RNA polymerase. The cleavage site is usually characterized by the presence of the base sequence AAUAAA near the cleavage site. After the mRNA has been cleaved, adenosine residues are added to the free 3’ end at the cleavage site.

[0184] As used herein, “transient” refers to expression of a non-integrated transgene for a period of hours, days or weeks, wherein the period of time of expression is less than the period of time for expression of the gene if integrated into the genome or contained within a stable plasmid replicon in the host cell. 46ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0185] As used herein, the terms “treat”, “treatment” and “treating” refer to the reduction or amelioration of the progression, severity and / or duration of a proliferative disorder, or the amelioration of one or more symptoms (preferably, one or more discernible symptoms) of a proliferative disorder resulting from the administration of one or more therapies (e.g., one or more therapeutic agents such as a CAR of the invention). In specific embodiments, the terms “treat”, “treatment” and “treating” refer to the amelioration of at least one measurable physical parameter of a proliferative disorder, such as growth of a tumor, not necessarily discernible by the patient. In some embodiments, the terms “treat”, “treatment” and “treating” -refer to the inhibition of the progression of a proliferative disorder, either physically by, e.g., stabilization of a discernible symptom, physiologically by, e.g., stabilization of a physical parameter, or both. In some embodiments, the terms “treat”, “treatment” and “treating” refer to the reduction or stabilization viral infection, number of cells infected with virus, or viral disorder symptoms.

[0186] The term “signal transduction pathway” refers to the biochemical relationship between a variety of signal transduction molecules that play a role in the transmission of a signal from one portion of a cell to another portion of a cell. The phrase “cell surface receptor” includes molecules and complexes of molecules capable of receiving a signal and transmitting signal across the membrane of a cell.

[0187] The term “subject” is intended to include living organisms in which an immune response can be elicited (e.g., mammals, human).

[0188] The term, a “substantially purified” cell refers to a cell that is essentially free of other cell types. A substantially purified cell also refers to a cell which has been separated from other cell types with which it is normally associated in its naturally occurring state. In some instances, a population of substantially purified cells refers to a homogenous population of cells. In other instances, this term refers simply to cell that have been separated from the cells with which they are naturally associated in their natural state. In some embodiments, the cells are cultured in vitro. In other embodiments, the cells are not cultured in vitro.

[0189] The term “therapeutic” as used herein means a treatment. A therapeutic effect is obtained by reduction, suppression, remission, or eradication of a disease state.

[0190] The term “prophylaxis” as used herein means the prevention of or protective treatment for a disease or disease state.

[0191] In the context of the present disclosure, “viral antigen” or “viral disorder antigen” or “antigen associated with a viral disorder” refer to antigens that are common to specific viral disorders. 47ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0192] The term “transfected” or “transformed” or “transduced” refers to a process by which exogenous nucleic acid is transferred or introduced into the host cell. A “transfected” or “transformed” or “transduced” cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny.

[0193] The term “specifically binds,” refers to an antibody, or a ligand, which recognizes and binds with a binding partner (e.g., a stimulatory tumor antigen) protein present in a sample, but which antibody or ligand does not substantially recognize or bind other molecules in the sample.

[0194] A subject “responds” to treatment if a parameter of a viral in the subject is retarded or reduced by a detectable amount, e.g., about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more as determined by any appropriate measure, e.g., by symptoms. In one example, a subject responds to treatment if the subject experiences a life expectancy extended by about 5%, 10%, 20%, 30%, 40%, 50% or more beyond the life expectancy predicted if no treatment is administered. In another example, a subject responds to treatment, if the subject has an increased disease-free survival, overall survival or increased time to progression. Several methods can be used to determine if a patient responds to a treatment including, for example, criteria provided by NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines®). For example, in the context of BALL, a complete response or complete responder, may involve one or more of: < 5% BM blast, >1000 neutrophil / ANC ( / µL). >100,000 platelets ( / µL) with no circulating blasts or extramedullary disease (no lymphadenopathy, splenomegaly, skin / gum infiltration / testicular mass / CNS involvement), Trilineage hematopoiesis, and no recurrence for 4 weeks. A partial responder may involve one or more of >50% reduction in BM blast, >1000 neutrophil / ANC ( / µL). >100,000 platelets ( / µL). A non-responder can show disease progression, e.g.,> 25% in BM blasts. In some embodiments, a complete responder is defined as having 7% or greater CD27+ CD45RO- cells in the CD8+ population. In some embodiments, the percent of CAR+ cells at pre- harvest levels distinguish responders (e.g., complete responders and partial responders) from non-responders (NR).

[0195] The term “relapse” as used herein refers to reappearance of a viral disorder after an initial period of responsiveness. The initial period of responsiveness may involve the level of symptoms or viral load falling below a certain threshold, e.g., below 20%, 1%, 10%, 5%, 4%, 3%, 2%, or 1%. The reappearance may involve the level symptoms or viral load rising above a certain threshold, e.g., above 20%, 1%, 10%, 5%, 4%, 3%, 2%, or 1%. In some embodiments, 48ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION the initial period of 30 responsiveness lasts at least 1, 2, 3, 4, 5, or 6 days; at least 1, 2, 3, or 4 weeks; at least 1, 2, 3, 4, 6, 8, 10, or 12 months; or at least 1, 2, 3, 4, or 5 years.

[0196] “Regulatable chimeric antigen receptor (RCAR),”as that term is used herein, refers to a set of polypeptides, typically two in the simplest embodiments, which when in a RCARX cell, provides the RCARX cell with specificity for a target cell, typically a virus infected cell, and with regulatable intracellular signal generation or proliferation, which can optimize an immune effector property of the RCARX cell. An RCARX cell relies at least in part, on an antigen binding domain to provide specificity to a target cell that comprises the antigen bound by the antigen binding domain. In some embodiments, an RCAR includes a dimerization switch that, upon the presence of a dimerization molecule, can couple an intracellular signaling domain to the antigen binding domain.

[0197] “Membrane anchor” or “membrane tethering domain”, as that term is used herein, refers to a polypeptide or moiety, e.g., a myristoyl group, sufficient to anchor an extracellular or intracellular domain to the plasma membrane.

[0198] “Switch domain,” as that term is used herein, e.g., when referring to an RCAR, refers to an entity, typically a polypeptide-based entity, that, in the presence of a dimerization molecule, associates with another switch domain. The association results in a functional coupling of a first entity linked to, e.g., fused to, a first switch domain, and a second entity linked to, e.g., fused to, a second switch domain. A first and second switch domain are collectively referred to as a dimerization switch. In some embodiments, the first and second switch domains are the same as one another, e.g., they are polypeptides having the same primary amino acid sequence, and are referred to collectively as a homodimerization switch. In some embodiments, the first and second switch domains are different from one another, e.g., they are polypeptides having different primary amino acid sequences and are referred to collectively as a heterodimerization switch. In some embodiments, the switch is intracellular. In some embodiments, the switch is extracellular. In some embodiments, the switch domain is a polypeptide-based entity, e.g., FKBP or FRB-based, and the dimerization molecule is small molecule, e.g., a rapalogue. In some embodiments, the switch domain is a polypeptide- based entity, e.g., an scFv that binds a myc peptide, and the dimerization molecule is a polypeptide, a fragment thereof, or a multimer of a polypeptide, e.g., a myc ligand or multimers of a myc ligand that bind to one or more myc scFvs. In some embodiments, the switch domain is a polypeptide-based entity, e.g., myc receptor, and the dimerization molecule is an antibody or fragments thereof, e.g., myc antibody. 49ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0199] “Dimerization molecule,” as that term is used herein, e.g., when referring to an RCAR, refers to a molecule that promotes the association of a first switch domain with a second switch domain. In some embodiments, the dimerization molecule does not naturally occur in the subject, or does not occur in concentrations that would result in significant dimerization. In some embodiments, the dimerization molecule is a small molecule, e.g., rapamycin or a rapalogue, e.g, RAD001.

[0200] The term “bioequivalent” refers to an amount of an agent other than the reference compound (e.g., RAD00I), required to produce an effect equivalent to the effect produced by the reference dose or reference amount of the reference compound (e.g., RAD001). In some embodiments the effect is the level of mTOR inhibition, e.g., as measured by P70 S6 kinase inhibition, e.g., as evaluated in an in vivo or in vitro assay, e.g., as measured by an assay described herein, e.g., the Boulay assay, or measurement of phosphorylated S6 levels by western blot. In some embodiments, the effect is alteration of the ratio of PD- I positive / PD- I negative T cells, as measured by cell sorting. In some embodiments a bioequivalent amount or dose of an mTOR inhibitor is the amount or dose that achieves the same level of P70 S6 kinase inhibition as does the reference dose or reference amount of a reference compound

[0201] In some embodiments, a bioequivalent amount or dose of an mTOR inhibitor is the amount or dose that achieves the same level of alteration in the ratio of PD- I positive / PD- I negative T cells as does the reference dose 20 or reference amount of a reference compound.

[0202] The term “low, immune enhancing, dose” when used in conjunction with an mTOR inhibitor, e.g., an allosteric mTOR inhibitor, e.g., RAD00I or rapamycin, or a catalytic mTOR inhibitor, refers to a dose of mTOR inhibitor that partially, but not fully, inhibits mTOR activity, e.g., as measured by the inhibition of P70 S6 kinase activity. Methods for evaluating Mtor activity, e.g., by inhibition of P70 S6 kinase, are discussed herein. The dose is insufficient to result in complete immune suppression but is sufficient to enhance the immune response. In some embodiments, the low, immune enhancing, dose of mTOR inhibitor results in a decrease in the number of PD-1 positive T cells and / or an increase in the number of PD-1 negative T cells, or an increase in the ratio of PD-1 negative T cells / PD- 1 positive T cells. In some embodiments, the low, immune enhancing, dose of mTOR inhibitor results in an increase in the number of nai:ve T cells. In some embodiments, the low, immune enhancing, dose of mTOR inhibitor results in one or more of the following: an increase in the expression of one or more of the following markers: CD62Lhigh, 5 CD127high, CD27+, and BCL2, e.g., on memory T cells, e.g., memory T cell precursors; a decrease in the expression of KLRGl, e.g., 50ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION on memory T cells, e.g., memory T cell precursors; and an increase in the number of memory T cell precursors, e.g., cells with any one or combination of the following characteristics: increased CD62Lhigh, increased CD127high,increased CD27+, decreased KLRG1, and increased BCL2; wherein any of the changes described above occurs, e.g., at least transiently, e.g., as compared to a non-treated subject.

[0203] Ranges: throughout this disclosure, various embodiments of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically 20 disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. As another example, a range such as 95-99% identity, includes something with 95%, 96%, 97%, 98% or 99% sequence identity, and includes subranges such as 96-99%, 96-98%, 96-97%, 97-99%, 97-98% and 98-99% sequence identity. This applies regardless of the breadth of the range.

[0204] The disclosure provides, at least in part, novel nucleic acid molecules encoding Chimeric Antigen Receptor (CAR) molecules comprising a first CAR comprising a peptide chosen from one of SEQ ID NO:1 through 200 or a variant thereof and a second CAR comprising a ligand to a costimulatory molecule or immunologically active protein. In some emodiments, the costimulatory molecule is CD28. The disclosure provides, at least in part, novel nucleic acid molecules encoding Chimeric Antigen Receptor (CAR) molecules comprising a first CAR comprising a EBV, SL9, Nef, or SARS targeting CAR and a second CAR comprising a ligand to a costimulatory molecule or immunologically active protein, e.g., dual CARs as described herein. In some embodiments, the first CAR comprises an antigen binding domain, and a first transmembrane domain; a first co-stimulatory signaling domain; and / or a first primary signaling domain. In some embodiments, the second CAR comprises an antigen binding domain, and a second transmembrane domain; a second co-stimulatory signaling domain; and / or a second primary signaling domain. In some embodiments of a CAR molecule disclosed herein, the CAR molecule comprises two identical polypeptide sequences, e.g., of a first and second transmembrane domain; a first and second co-stimulatory domain; and / or a first and second primary signaling domain, the polypeptide sequences of which are encoded by different nucleotide sequences. Also disclosed herein are methods of using said 51ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION CAR molecules. In some embodiments, the disclosure relates to nucleic acid molecules encoding a monovalent CAR comprising either the first CAR or the second CAR

[0205] Without wishing to be bound by theory, it is believed that in some embodiments, a nucleic acid molecule encoding a CAR molecule, e.g., a dual CAR molecule, is optimized, e.g., codon optimized, to prevent recombination, e.g., homologous recombination. In some embodiments, a CAR molecule, e.g., a dual CAR molecule, comprises two domains, e.g., a first transmembrane domain and a second transmembrane domain, each of which comprises a similar amino acid sequence but is encoded by a different nucleotide sequence.

[0206] In some embodiments, a CAR molecule disclosed herein comprises a first CAR comprising a first antigen binding domain which binds a first transmembrane domain; a first co- stimulatory signaling domain; and / or a first primary signaling domain.

[0207] In some embodiments, the first antigen binding domain comprises one or more (e.g., all three) light chain complementarity determining region I (LC CDR1), light chain complementarity determining region 2 (LC CDR2), and light chain complementarity determining region 3 (LC CDR3) of a binding domain described herein; and / or one or more (e.g., all three) heavy chain complementarity determining region 1 (HC CDR1 ), heavy chain complementarity determining region 2 (HC CDR2), and heavy chain complementarity determining region 3 (HC CDR3) of a binding domain described herein, In some embodiments, the antigen binding domain comprises a LC CDRI, LC CDR2 and LC CDR3 of a binding domain described herein; and / or a HC CDR1, HC CDR2 and HC CDR3 of a binding domain described herein.

[0208] In some embodiments, a CAR molecule disclosed herein comprises a first CAR comprising a first transmembrane domain and a second CAR comprising a second transmembrane domain. In some embodiments, the first transmembrane domain and the second transmembrane domain comprise the same amino acid sequence, e.g., as disclosed herein. In some embodiments, the first transmembrane domain and the second transmembrane domain are encoded by a first nucleotide sequence and a second nucleotide sequence, respectively. In some embodiments, the first nucleotide sequence and the second nucleotide sequence differ by at least one nucleotide.

[0209] In some embodiments, the first transmembrane domain and the second transmembrane domain are the same transmembrane domain, e.g., chosen from the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD4S, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CDIS4. 52ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0210] In some embodiments, the first transmembrane domain and the second transmembrane domain are different transmembrane domains, e.g., chosen from the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD4S, CD4, CDS, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CDIS4.

[0211] In some embodiments, a nucleic acid molecule encoding a CAR molecule described herein comprises a first CAR comprising a first transmembrane domain and a second CAR comprising a second transmembrane domain. In some embodiments, the first transmembrane domain and the second transmembrane domain comprise the CD8 alpha transmembrane domain. In some embodiments, the first transmembrane domain and the second transmembrane domain comprise the amino acid sequence of SEQ ID NO: 5 or an amino acid sequence with at least about 90% identity thereto.

[0212] In some embodiments, a nucleotide sequence that encodes the first transmembrane domain and is comprised in the nucleic acid molecule is different from a nucleotide sequence that encodes the second transmembrane domain and is comprised in the nucleic acid molecule.

[0213] In some embodiments, a CAR molecule disclosed herein comprises a first CAR comprising a first co-stimulatory domain and a second CAR comprising a second co- stimulatory domain. In some embodiments, the first co-stimulatory domain and the second co- stimulatory domain comprise the same amino acid sequence, e.g., as disclosed herein. In some embodiments, the first co-stimulatory domain and the second co-stimulatory domain are encoded by a first nucleotide sequence and a second nucleotide sequence, respectively. In some embodiments, the first nucleotide sequence and the second nucleotide sequence differ by at least one nucleotide.

[0214] In some embodiments, the first co-stimulatory domain and the second co- stimulatory domain are the same co-stimulatory domain, e.g., chosen from a signaling domain of OX40, CD2, CD27, CD28, CD5, ICAM-1, LFA-1 (CD1 la / CD18), ICOS (CD278) or 4- lBB (CD137).

[0215] In some embodiments, the first co-stimulatory domain and the second co- stimulatory domain are different co-stimulatory domains, e.g., chosen from a signaling domain of OX40, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD1 la / CD18), COS (CD278) or 4- lBB (CD137).

[0216] In some embodiments, a nucleic acid molecule encoding a CAR molecule described herein comprises a first CAR comprising a first co-stimulatory domain and a second CAR comprising a second co-stimulatory domain. In some embodiments, the first co-stimulatory domain and the second co-stimulatory domain comprise a 4-lBB co-stimulatory domain. 53ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0217] In some embodiments, a nucleotide sequence that encodes the first co-stimulatory domain and is comprised in the nucleic acid molecule is different from a nucleotide sequence that encodes the second co-stimulatory domain and is comprised in the nucleic acid molecule.

[0218] In some embodiments, the present disclosure provides a nucleic acid molecule encoding a CAR molecule, e.g., comprising (i) a first CAR comprising a NYESO-1 antigen binding domain and (ii) a second CAR comprising a second antigen binding domain. In some embodiments, the nucleic acid comprises RNA or DNA In some embodiments, the nucleic acid sequences encoding (i) and (ii) are situated in the same orientation, e.g., transcription of the nucleic acid sequences encoding (i) and (ii) proceeds in the same direction. In some embodiments, the nucleic acid sequences encoding (i) and (ii) are situated in different orientations. In some embodiments, a single promoter controls expression of the nucleic acid sequences encoding (i) and (ii). In some embodiments, a nucleic acid encoding a protease cleavage site (such as a T2A, P2A, E2A, or F2A cleavage site) is situated between the nucleic acid sequences encoding (i) and (ii). In some embodiments, the protease cleavage site is placed such that a cell can express a fusion protein comprising (i) and (ii), which protein is subsequently processed into two peptides by proteolytic cleavage. In some embodiments, the nucleic acid sequences encoding (i) is upstream of the nucleic acid sequences encoding (ii), or the nucleic acid sequences encoding (ii) is upstream of the nucleic acid sequences encoding (i). In some embodiments, a first promoter controls expression of the nucleic acid sequence encoding (i) and a second promoter controls expression of the nucleic acid sequence encoding (ii). In some embodiments, the nucleic acid is a plasmid. In some embodiments, the nucleic acid comprises a viral packaging element. In some embodiments, the present disclosure provides a cell, e.g., an immune effector cell, comprising the nucleic acid described herein, e.g., a nucleic acid comprising (i) and as described above. The cell may comprise a protease (e.g., endogenous or exogenous) that cleaves a T2A, P2A, E2A, or F2A cleavage site.

[0219] Exemplary nucleotide and amino acid sequences of a CAR molecule, e.g., dual CAR molecule or a monovalent CAR molecule disclosed herein are provided in Tables 1A and 1B. In some embodiments, the disclosure relates to an effector cell comprising any one or combination of the amino acid sequences of Table 1A or any one or combination of the nucleic acid sequences of Table 1B. Table 1A: Single and Dual CAR amino acid sequences Identifier SEQ ID Sequence5ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION Antigen 201 GTHDKCENPKX1X2X3X4X5X6X7X8, where binding X1X2X3X4X5X6X7X8is one of SEQ ID NOS: 1 domain 1 through 200 FT is V 1 Y AN , KE NITable 1B: Single and Dual CAR nucleic acid sequences Identifier SEQ ID NO. Nucleic Acid Sequence55ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION Antigen490ggcacccacgacaagtgcgagaaccccaagnnnnnnnnnnnnnnnnnnnnnnnnbinding where nnnnnnnnnnnnnnnnnnnnnnnn is one of SEQ ID NOS: 290– domain 1 489 nca ac 0– cagag S: n)11tg( cgt ac 14gtgctg ac

[0220] Table 2 provides nucleotide and amino acid sequences for additional CAR components, e.g., signal peptide, linkers and P2A sites. Table 2: Additional CAR components Identifier SEQ SequenceATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION Signal peptide 1 256 atggccctccctgtcaccgccctgctgcttccgctggctcttctgctccacgccgctcggcc c 206 MALPVTALLLPLALLLHAARP gacggc ccggtctgctt cttt L ct c c L c c ct A57ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION CD8 hinge and 221 accaccacccctgcgcctcggcctcctaccccggctcccactatcgcgagccagccgct transmembrane ga domain 4 gcctgcggcctgaggcttgccgaccggccgctggcggcgccgtgcatactcggggcct ctg L ag tgc a t ca gat caagcg G a a t g a K I P58ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION CD3zeta 233 agagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaacca Sequence 2 gct ctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtg t g g K I P ca cg ct g ca KR gctgg aca aaa gat KR59ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION Linker 242 GGGGSGGGGSGGGGS between scFVs 3 accccp q g g q the nucleic acid sequences encoding each. SEQ Amino Acid Sequence SEQ Nucleic Acid Sequence ID ID60ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 34 EWFETYVA 323 gagtggttcgagacctacgtggcc 35 EWFWIAHD 324 gagtggttctggatcgcccacgac 36 EWFYTYVH 325 gagtggttctacacctacgtgcacATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 92 YMNLNNFG 381 tacatgaacctgaacaacttcggc 93 QYELSNQM 382 cagtacgagctgagcaaccagatg 94 YMNLYNIG 383 tacatgaacctgtacaacatcggcATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 150 WTNNFGKT 439 tggaccaacaacttcggcaagacc 151 KDHGIIVA 440 aaggaccacggcatcatcgtggcc 152 AHEWWTHN 441 gcccacgagtggtggacccacaac, molecules comprising one or more nucleic acid sequences encoding an antibody or antibody fragment comprising a first binding domain and a second binding domain, the first binding 63ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION domain comprising a peptide that binds to CD3 and a second binding domain that binds to a viral antigen, wherein the second binding domain comprises a peptide comprises an amino acid sequence chosen from SEQ ID NOS: 1 through 202 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 1 through 202. In some embodiments, the one or more nucleic acid sequences comprise the sequence of one or more of SEQ ID NOS: 290–489, or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 290–489. In some embodiments, the antibody or antibody fragment is a bispecific T-cell engager (or BiTE) comprising a first binding domain comprising a peptide that binds to CD3 and a second binding domain that binds to a viral antigen, wherein the second binding domain comprises a peptide comprises an amino acid sequence chosen from SEQ ID NO: 1 through 202 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NO: 1 through 202. Although not bound by theory, the purpose of the BiTE is to transiently associate or link a CD-3 expressing T cell associated with the first binding region and a cancer cell expressing a tumor associated antigen associated with the second binding region. Therefore the BiTE bridges a transient binding connection between the CD-3 expressing cell and a cancer target cell, thereby inducing destruction of the cancer cell. In some embodiments, the BiTE comprises a [TRACeR〗_(MHC-I,A02)^(cancer antigen binding domain)- antiCD3 BiTE comprising a first binding domain comprising a peptide binding CD3 and a second binding domain comprising one or more amino acid sequence chosen from SEQ ID NOS: 1 through 205 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 1 through 205. In some embodiments, the one or more nucleic acid sequences comprise the sequence of one or more of SEQ ID NOS: 290– 494, or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 290–494. In some embodiments, the second binding region comprises a scaffold domain, a portion of an MHC Class I or Class II molecule, and an amino acid sequence disclosed herein positioned contiguously or non-contiguously within or fused to the scaffold domain and the portion of the MHC Class I or Class II domain. In some embodiments, the disclosure relates to a composition comprising the one or more nucleic acid molecules. In some 64ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION embodiments, the disclosure relates to a pharmaceutical composition comprising the one or more nucleic acid molecules. In some embodiments, the one or more nucleic acid sequences encode the first binding domain comprising a peptide that binds to CD3. In some embodiments, the peptide that binds to CD3 is one disclosed in U.S. Patent No. 9,688,760 (“Anti-leukocyte adhesion for the mitigation of potential adverse events caused by CD3-specific binding domains”); 8,247,194 (“Preparation of scFv antibody fragments”); or 7,235,641 (“Bispecific antibodies”), which are incorporated herein by reference in their entireties. In some embodiments, the one or more nucleic acid sequence encodes the peptide that binds to CD3 disclosed in the U.S. Patent Nos. 9,688,760; 8,247,194; or 7,235,641. In some embodiments, the peptide that binds to CD3 comprises one or more anti-CD3 CDRs. In some embodiments, the one or more anti-CD3 CDRs comprise one or more anti-CD3 heavy chain CDR. In some embodiments, the one or more anti-CD3 CDRs comprise one or more anti-CD3 light chain CDR. In some embodiments, the one or more anti-CD3 CDRs comprise one or more anti-CD3 heavy chain CDR and one or more anti-CD3 light chain CDR. In some embodiments, the one or more anti-CD3 heavy chain CDR comprise one or more of CD3 CDR-H1 of SEQ ID NO: 503 (RYTMH) or SEQ ID NO: 504 (GYTFTRYTMH), CD3 CDR-H2 of SEQ ID NO: 505 (YINPSRGYTNYNQKFKD) or CD3 CDR-H3 of SEQ ID NO: 506 (YYDDHYCLDY), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 503 through 506. In some embodiments, the one or more anti-CD3 light chain CDR comprise one or more of CD3 CDR-L1 of SEQ ID NO: 507 (RASSSVSYMN), CD3 CDR-L2 of SEQ ID NO: 508 (DTSKVAS) and CD3 CDR-L3 of SEQ ID NO: 509 (QQWSSNPLT), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 507 through 509. In some embodiments, the one or more nucleic acid sequences encoding the peptide that binds CD-3 encode a heavy chain amino acid sequence comprising the sequence DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRG YTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG TTLTVSS (SEQ ID NO: 510), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 510. In some embodiments, the one or more nucleic acid sequences encoding the heavy chain comprise gatatcaaactgcagcagtcaggggctgaactggcaagacctggggcctcagtgaagatgtcctgcaagacttctggctacaccttta 65ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION ctaggtacacgatgcactgggtaaaacagaggcctggacagggtctggaatggattggatacattaatcctagccgtggttatactaat tacaatcagaagttcaaggacaaggccacattgactacagacaaatcctccagcacagcctacatgcaactgagcagcctgacatctg aggactctgcagtctattactgtgcaagatattatgatgatcattactgccttgactactggggccaaggcaccactctcacagtctcctc a (SEQ ID NO: 511), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 511. In some embodiments, one or more nucleic acid sequences encoding the peptide that binds CD-3 encode a light chain amino acid sequence comprising the sequence DIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGV PYRFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK (SEQ ID NO: 512), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 512. In some embodiments, the one or more nucleic acid sequences encoding the light chain comprise gacattcagctgacccagtctccagcaatcatgtctgcatctccaggggagaaggtcaccatgacctgcagagccagttcaagtgtaa gttacatgaactggtaccagcagaagtcaggcacctcccccaaaagatggatttatgacacatccaaagtggcttctggagtcccttat cgcttcagtggcagtgggtctgggacctcatactctctcacaatcagcagcatggaggctgaagatgctgccacttattactgccaaca gtggagtagtaacccgctcacgttcggtgctgggaccaagctggagctgaaa (SEQ ID NO: 513), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 513. In some embodiments, the disclosure relates to a composition or pharmaceutical composition comprising the one or more nucleic acid molecules. In some embodiments, the composition or pharmaceutical composition further comprises one or more nucleic acid sequences encoding a peptide that binds to CD-19. In some embodiments, the peptide that binds to CD-19 comprises one or more anti-CD-19 CDRs. In some embodiments, the one or more anti-CD19 CDRs comprise one or more anti-CD19 heavy chain CDR. In some embodiments, the one or more anti-CD19 CDRs comprise one or more anti-CD19 light chain CDR. In some embodiments, the one or more anti-CD19 CDRs comprise one or more anti-CD19 heavy chain CDR and one or more anti-CD19 light chain CDR. In some embodiments, the one or more anti-CD19 heavy chain CDR comprise one or more of CD19 CDR-H1 of SEQ ID NO: 514 (SYWMN) or SEQ ID NO: 515 (GYAFSSYWMN), CD19 CDR-H2 of SEQ ID NO: 516 (QIWPGDGDTNYNGKFKG) and CD19 CDR-H3 of SEQ ID NO: 517 (RETTTVGRYYYAMDY), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 514 through 517. In some embodiments, the 66ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION one or more anti-CD19 light chain CDR comprise one or more of CD19 CDR-L1 of SEQ ID NO: 518 (KASQSVDYDGDSYLN), CD19 CDR-L2 of SEQ ID NO: 519 (DASNLVS) and CD19 CDR-L3 of SEQ ID NO: 520 (QQSTEDPWT), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 518 through 520.

[0223] In some embodiments, the disclosure also relates to compositions and pharmaceutical compositions comprising an antibody or antibody fragment comprising a first binding domain and a second binding domain, the first binding domain comprising a peptide that binds to CD3 and a second binding domain that binds to a viral antigen, wherein the second binding domain comprises a peptide comprises an amino acid sequence chosen from SEQ ID NO: 1 through 202 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NO: 1 through 202. In some embodiments, the antibody or antibody fragment is a bispecific T-cell engager (or BiTE) comprising a first binding domain comprising a peptide that binds to CD3 and a second binding domain that binds to a viral antigen, wherein the second binding domain comprises a peptide comprises an amino acid sequence chosen from SEQ ID NO: 1 through 202 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NO: 1 through 202. Although not bound by theory, the purpose of the BiTE is to transiently associate or link a CD-3 expressing T cell associated with the first binding region and a cancer cell expressing a tumor associated antigen associated with the second binding region. Therefore the BiTE bridges a transient binding connection between the CD-3 expressing cell and a cancer target cell, thereby inducing destruction of the cancer cell. In some embodiments, the BiTE comprises a [TRACeR〗_(MHC-I,A02)^(cancer antigen binding domain)- antiCD3 BiTE comprising a first binding domain comprising a peptide binding CD3 and a second binding domain comprising one or more amino acid sequence chosen from SEQ ID NO: 1 through 205 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 1 through 205. In some embodiments, the second binding region comprises a scaffold domain, a portion of an MHC Class I or Class II molecule, and an amino acid sequence disclosed herein positioned contiguously or non- contiguously within or fused to the scaffold domain and the portion of the MHC Class I or Class II domain. In some embodiments, the peptide that binds to CD3 is one disclosed in U.S. 67ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION Patent No.9,688,760 (“Anti-leukocyte adhesion for the mitigation of potential adverse events caused by CD3-specific binding domains”); 8,247,194 (“Preparation of scFv antibody fragments”); or 7,235,641 (“Bispecific antibodies”), which are incorporated herein by reference in their entireties. In some embodiments, the peptide that binds to CD3 comprises one or more anti-CD3 CDRs. In some embodiments, the one or more anti-CD3 CDRs comprise one or more anti-CD3 heavy chain CDR. In some embodiments, the one or more anti-CD3 CDRs comprise one or more anti-CD3 light chain CDR. In some embodiments, the one or more anti-CD3 CDRs comprise one or more anti-CD3 heavy chain CDR and one or more anti- CD3 light chain CDR. In some embodiments, the one or more anti-CD3 heavy chain CDR comprise one or more of CD3 CDR-H1 of SEQ ID NO: 503 (RYTMH) or SEQ ID NO: 504 (GYTFTRYTMH), CD3 CDR-H2 of SEQ ID NO: 505 (YINPSRGYTNYNQKFKD) or CD3 CDR-H3 of SEQ ID NO: 506 (YYDDHYCLDY), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 503 through 506. In some embodiments, the one or more anti-CD3 light chain CDR comprise one or more of CD3 CDR- L1 of SEQ ID NO: 507 (RASSSVSYMN), CD3 CDR-L2 of SEQ ID NO: 508 (DTSKVAS) and CD3 CDR-L3 of SEQ ID NO: 509 (QQWSSNPLT), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 506 through 508. In some embodiments, the peptide that binds CD-3 comprises a heavy chain amino acid sequence comprising the sequence DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRG YTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG TTLTVSS (SEQ ID NO: 510), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 510. In some embodiments, the peptide that binds CD-3 comprises a light chain amino acid sequence comprising the sequence DIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGV PYRFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK (SEQ ID NO: 511), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 511. In some embodiments, the composition or pharmaceutical composition further comprises a peptide that binds to CD-19. In some embodiments, the peptide that binds to CD-19 comprises one or more anti-CD-19 CDRs. In some embodiments, the one or more anti-CD19 CDRs 68ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION comprise one or more anti-CD19 heavy chain CDR. In some embodiments, the one or more anti-CD19 CDRs comprise one or more anti-CD19 light chain CDR. In some embodiments, the one or more anti-CD19 CDRs comprise one or more anti-CD19 heavy chain CDR and one or more anti-CD19 light chain CDR. In some embodiments, the one or more anti-CD19 heavy chain CDR comprise one or more of CD19 CDR-H1 of SEQ ID NO: 512 (SYWMN) or SEQ ID NO: 513 (GYAFSSYWMN), CD19 CDR-H2 of SEQ ID NO: 514 (QIWPGDGDTNYNGKFKG) and CD19 CDR-H3 of SEQ ID NO: 515 (RETTTVGRYYYAMDY), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 512 through 515. In some embodiments, the one or more anti-CD19 light chain CDR comprise one or more of CD19 CDR-L1 of SEQ ID NO: 516 (KASQSVDYDGDSYLN), CD19 CDR-L2 of SEQ ID NO: 517 (DASNLVS) and CD19 CDR-L3 of SEQ ID NO: 518 (QQSTEDPWT), or a functional variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to the respective one of SEQ ID NOS: 516 through 518.

[0224] BiTE therapies and manufacturing the same are known in the art but may be as disclosed in Slaney, CY et al., Cancer Discov.8(8):924-934 (2018), Ellerman, D. Methods, 154(1): 102-117 (2019), and Vafa, O. et al. Forehead. Oncol.15 April 2020; doi.org / 10.3389 / fonc.2020.00446, the contents of each of which are incorporated by reference in their entireties. See also U.S. Patent Nos.12,258,417 and 11,661,462, which are incorporated herein by reference in its entirety.

[0225] In some embodiments, the disclosure relates to a composition or pharmaceutical composition comprising an antibody or antibody fragment herein or a or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity thereto. In some embodiments, the disclosure relates to a composition or pharmaceutical composition comprising one or more nucleic acid molecule comprising one or more nucleic acid sequence encoding an antibody or antibody fragment herein or a or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity thereto. In some embodiments, the disclosure relates to compositions and pharmaceutical compositions comprising a dual CAR herein or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity thereto.

[0226] In some embodiments, the disclosure relates to compositions and pharmaceutical compositions comprising one more nucleic acid molecule comprising one or more nucleic acid 69ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION sequences encoding a dual CAR herein or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity thereto. In some embodiments, the disclosure relates to compositions and pharmaceutical compositions comprising a monovalent CAR herein or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity thereto. In some embodiments, the disclosure relates to compositions and pharmaceutical compositions comprising one or more nucleic acid molecules comprising one or more nucleic acid sequences encoding a monovalent CAR herein or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity thereto.

[0227] In some embodiments, an antibody or antibody fragment herein comprises one or more amino acid sequence comprising the sequence of SEQ ID NO: 1 through 200 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 1 through 200. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are contiguous. In some embodiments, each sequence selected from SEQ ID NO: 1 through 200 is contiguous in that there is no amino acid intervening between any of the eight amino acids. In some embodiments, at least one sequence selected from SEQ ID NO: 1 through 200 is non- contiguous by having a spacer of one or more amino acid inserted between two of residues thereof. Some embodiments comprise one or more nucleic acid sequence encoding an antibody or antibody fragment herein. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid 70ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION sequence of 1 through 202 or a variant thereof is contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202.

[0228] In some embodiments, a BiTE herein comprises one or more amino acid sequence comprising the sequence of SEQ ID NO: 1 through 200 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 1 through 200. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are contiguous. In some embodiments, each sequence selected from SEQ ID NO: 1 through 200 is contiguous in that there is no amino acid intervening between any of the eight amino acids. In some embodiments, at least one sequence selected from SEQ ID NO: 1 through 200 is non- contiguous by having a spacer of one or more amino acid inserted between two of residues thereof. Some embodiments, comprise one or more nucleic acid sequence encoding a BiTE herein. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 71ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 202. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202.

[0229] In some embodiments, a CAR herein comprises one or more amino acid sequence comprising the sequence of SEQ ID NO: 1 through 200 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 1 through 200. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are contiguous. In some embodiments, each sequence selected from SEQ ID NO: 1 through 200 is contiguous in that there is no amino acid intervening between any of the eight amino acids. In some embodiments, at least one sequence selected from SEQ ID NO: 1 through 200 is non- contiguous by having a spacer of one or more amino acid inserted between two of residues thereof. Some embodiments comprise one or more nucleic acid sequence encoding a CAR herein. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the 72ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202.

[0230] In some embodiments, of a dual CAR herein, at least one or both of the CARs comprises one or more amino acid sequence comprising the sequence of SEQ ID NO: 1 through 200 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 1 through 200. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are contiguous. In some embodiments, each sequence selected from SEQ ID NO: 1 through 200 is contiguous in that there is no amino acid intervening between any of the eight amino acids. In some embodiments, at least one sequence selected from SEQ ID NO: 1 through 200 is non-contiguous by having a spacer of one or more amino acid inserted between two of residues thereof. Some embodiments comprise one or more nucleic acid sequence encoding a dual CAR herein. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202. 73ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0231] In some embodiments, of a tandem CAR herein, at least one or both of the CARs comprises one or more amino acid sequence comprising the sequence of SEQ ID NO: 1 through 200 or a variant thereof having at least about 70%, about 72%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% sequence identity to SEQ ID NO: 1 through 200. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are non-contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, the two amino acid sequences are contiguous. In some embodiments, where the one or more amino acid sequence comprises two or more amino acid sequences, at least two of the amino acid sequences are contiguous. In some embodiments, each sequence selected from SEQ ID NO: 1 through 200 is contiguous in that there is no amino acid intervening between any of the eight amino acids. In some embodiments, at least one sequence selected from SEQ ID NO: 1 through 200 is non-contiguous by having a spacer of one or more amino acid inserted between two of residues thereof. Some embodiments comprise one or more nucleic acid sequence encoding a tandem CAR herein. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the first nine amino acids of SEQ ID NOS: 201, 205, or 202, in the orientation from amino to carboxy terminal of the first nine amino acids of SEQ ID NOS: 201, 205, or 202 and then the sequence of 1 through 202 or a variant thereof. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202. In some embodiments, the amino acid sequence of 1 through 202 or a variant thereof is non-contiguous with the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202 in the orientation from amino to carboxy terminal of the sequence of 1 through 202 or a variant thereof then the QNL SEQ ID NO: 205 or the QNX of SEQ ID NO: 202. Tandem CARs 74ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0232] In some embodiments, disclosed herein are CARs comprising a bispecific antigen binding domain, e.g., tandem CARs. In some embodiments, a bispecific antigen binding domain comprises two antigen binding domains, e.g., a first antigen binding domain and a second antigen binding domain. In some embodiments of the bispecific antigen binding domain, the first antigen binding domain is an antibody molecule, e.g., an antibody binding domain (e.g., a scFv). In some embodiments of the bispecific antigen binding domain, the second antigen binding domain is an antibody molecule, e.g., an antibody binding domain (e.g., a scFv). Within each antibody molecule, e.g., scFv, of the bispecific antigen binding domain, the VH can be upstream or downstream of the VL.

[0233] In some embodiments, the upstream antibody or antibody fragment (e.g., scFv) is arranged with its VH (VHI) upstream of its VL (VLI) and the downstream antibody or antibody fragment (e.g., scFv) is arranged with its VL (VL2) upstream of its VH (VH2), such that the overall bispecific antibody molecule has the arrangement VH1-VL1-VL2-VH2, from an N- to C- terminal orientation.

[0234] In some embodiments, the upstream antibody or antibody fragment (e.g., scFv) is arranged with its VL (VLI) upstream of its VH (VHI) and the downstream antibody or antibody fragment (e.g., scFv) is arranged with its VH (VH2) upstream of its VL (VL2), such that the overall bispecific antibody molecule has the arrangement VL1-VH1-VH2-VL2, from an N- to C- terminal orientation.

[0235] In some embodiments, the upstream antibody or antibody fragment (e.g., scFv) is arranged with its VL (VLI) upstream of its VH (VHI) and the downstream antibody or antibody fragment (e.g., scFv) is arranged with its VL (VL2) upstream of its VH (VH2), such that the overall bispecific antibody molecule has the arrangement VL1-VH1-VL2-VH2, from an N- to C- terminal orientation. In yet other embodiments, the upstream antibody or antibody fragment (e.g., scFv) is arranged with its VH (VHI) upstream of its VL (VLI) and the downstream antibody or antibody fragment (e.g., scFv) is arranged with its VH (VH2) upstream of its VL (VL2), such that the overall bispecific antibody molecule has the arrangement VH1-VL1-VH2- VL2, from an N- to C- terminal orientation.

[0236] In any of the aforesaid configurations, optionally, a linker is disposed between the two antibodies or antibody fragments (e.g., scFvs), e.g., between VLI and VL2 if the construct is arranged as VH1-VL1-VL2-VH2; between VHI and VH2 if the construct is arranged as VLI- VH1-VH2-VL2; between VHI and VL2 if the construct is arranged as VL1-VH1-VL2-VH2; or between VLl and VH2 if the construct is arranged as VH1-VL1-VH2-VL2. In general, the linker between the two scFvs should be long enough to avoid mispairing between the 75ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION domains of the two scFvs. The linker may be a linker as described herein. In some embodiments, the linker is a (Gly4-Ser)n linker (SEQ ID NO: 251), wherein n is 1, 2, 3, 4, 5, or 6. In some embodiments, the linker is (Gly4-Ser)n (SEQ ID NO: 249), wherein n = 1, e.g., the linker has the amino acid sequence Gly4-Ser (SEQ ID NO: 249). In some embodiments, the linker is (Gly4-Ser)n, wherein n= 4 (SEQ ID NO: 250). In some embodiments, the linker comprises, e.g., consists of, the amino acid sequence: LAEAAAK (e.g., SEQ ID NO: 240).

[0237] In any of the aforesaid configurations, optionally, a linker is disposed between the VL and VH of the first scFv. Optionally, a linker is disposed between the VL and VH of the second scFv. In constructs that have multiple linkers, any two or more of the linkers can be the same or different. Accordingly, in some embodiments, a bispecific CAR comprises VLs, VHs, and optionally one or more linkers in an arrangement as described herein.

[0238] In some embodiments, each antibody molecule, e.g., each antigen binding domain (e.g., each scFv) comprises a linker between the VH and the VL regions. In some embodiments, the linker between the VH and the VL regions is a (Gly4-Ser)n linker (SEQ ID NO: 251), wherein n is 1, 2, 3, 4, 5, or 6. In some embodiments, the linker is (Gly4-Ser)n (SEQ ID NO: 249), wherein n = 1, e.g., the linker has the amino acid sequence Gly4-Ser. In some embodiments, the linker is (Gly4-Ser)n (SEQ ID NO: 249), wherein n= 4 (SEQ ID NO: 250). In some embodiments, the VH and VL regions are connected without a linker. Split CAR

[0239] In some embodiments, the CAR-expressing cell uses a split CAR. The split CAR approach is described in more detail in PCT publications WO2014 / 055442 and WO2014 / 055657, incorporated herein by reference. Briefly, a split CAR system comprises a cell expressing a first CAR having a first antigen binding domain and a costimulatory domain (e.g., 4-lBB), and the cell also expresses a second CAR having a second antigen binding domain and an intracellular signaling domain (e.g., CD3 zeta). When the cell encounters the first antigen, the costimulatory domain is activated, and the cell proliferates. When the cell encounters the second antigen, the intracellular signaling domain is activated and cell-killing activity begins. Thus, the CAR-expressing cell is only fully activated in the presence of both antigens. RNA Transfection

[0240] Disclosed herein are methods for producing an in vitro transcribed RNA CAR. The present disclosure also includes a CAR encoding RNA construct that can be directly transfected into a cell. A method for generating mRNA for use in transfection can involve in vitro 76ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION transcription (IVT) of a template with specially designed primers, followed by polyA addition, to produce a construct containing 3’ and 5’ untranslated sequence (“UTR”), a 5’ cap and / or Internal Ribosome Entry Site (IRES), the nucleic acid to be expressed, and a polyA tail, typically 50- 2000 bases in length. RNA so produced can efficiently transfect different kinds of cells. In some embodiments, the template includes sequences for the CAR.

[0241] In one embodiment the CAR, e.g., dual CAR or tandem CAR, is encoded by a messenger RNA (mRNA). In one embodiment the mRNA encoding the CAR, e.g., dual CAR or tandem CAR, is introduced into an immune effector cell, e.g., a T cell or a NK cell, for production of a CAR- expressing cell, e.g., a CART cell or a CAR NK cell.

[0242] In some embodiments, the in vitro transcribed RNA of a CAR can be introduced to a cell as a form of transient transfection. The RNA is produced by in vitro transcription using a polymerase chain reaction (PCR)-generated template. DNA of interest from any source can be directly converted by PCR into a template for in vitro mRNA synthesis using appropriate primers and RNA polymerase. The source of the DNA can be, for example, genomic DNA, plasmid DNA, phage DNA, cDNA, synthetic DNA sequence or any other appropriate source of DNA.

[0243] The desired temple for in vitro transcription is a CAR of the present disclosure. For example, the template for the RNA CAR comprises an extracellular region comprising a single chain variable domain of an anti-tumor antibody; a hinge region, a transmembrane domain (e.g., a transmembrane domain of CD8a); and a cytoplasmic region that includes an intracellular signaling domain, e.g., comprising the signaling domain of CD3-zeta and the signaling domain of 4-lBB.

[0244] In some embodiments, the DNA to be used for PCR contains an open reading frame. The DNA can be from a naturally occurring DNA sequence from the genome of an organism. In some embodiments, the nucleic acid can include some or all of the 5’ and / or 3’ untranslated regions (UTRs). The nucleic acid can include exons and introns. In some embodiments, the DNA to be used for PCR is a human nucleic acid sequence. In some embodiments, the DNA to be used for PCR is a human nucleic acid sequence including the 5’ and 3’ UTRs. The DNA can alternatively be an artificial DNA sequence that is not normally expressed in a naturally occurring organism. An exemplary artificial DNA sequence is one that contains portions of genes that are ligated together to form an open reading frame that encodes a fusion protein. The portions of DNA that are ligated together can be from a single organism or from more than one organism. 77ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0245] PCR is used to generate a template for in vitro transcription of mRNA which is used for transfection. Methods for performing PCR are well known in the art. Primers for use in PCR are designed to have regions that are substantially complementarity to regions of the DNA to be used as a template for the PCR. “Substantially complementarity,” as used herein, refers to sequences of nucleotides where a majority or all of the bases in the primer sequence are complementarity, or one or more bases are non-complementarity, or mismatched. Substantially complementarity sequences are able to anneal or hybridize with the intended DNA target under annealing conditions used for PCR. The primers can be designed to be substantially complementarity to any portion of the DNA template. For example, the primers can be designed to amplify the portion of a nucleic acid that is normally transcribed in cells (the open reading frame), including 5’ and 3’ UTRs. The primers can also be designed to amplify a portion of a nucleic acid that encodes a particular domain of interest. In some embodiments, the primers are designed to amplify the coding region of a human cDNA, including all or portions of the 5’ and 3’ UTRs. Primers useful for PCR can be generated by synthetic methods that are well known in the art. “Forward primers” are primers that contain a region of nucleotides that are substantially complementarity to nucleotides on the DNA template that are upstream of the DNA sequence that is to be amplified. “Upstream” is used herein to refer to a location 5, to the DNA sequence to be amplified relative to the coding strand. “Reverse primers” are primers that contain a region of nucleotides that are substantially complementarity to a double-stranded DNA template that are downstream of the DNA sequence that is to be amplified. “Downstream” is used herein to refer to a location 3’ to the DNA sequence to be amplified relative to the coding strand.

[0246] Any DNA polymerase useful for PCR can be used in the methods disclosed herein. The reagents and polymerase are commercially available from a number of sources.

[0247] Chemical structures with the ability to promote stability and / or translation efficiency may also be used. The RNA preferably has 5’ and 3’ UTRs. In some embodiments, the 5’ UTR is between one and 3000 nucleotides in length. The length of 5’ and 3’ UTR sequences to be added to the coding region can be altered by different methods, including, but not limited to, designing primers for PCR that anneal to different regions of the UTRs. Using this approach, one of ordinary skill in the art can modify the 5’ and 3’ UTR lengths required to achieve optimal translation efficiency following transfection of the transcribed RNA.

[0248] The 5’ and 3’ UTRs can be the naturally occurring, endogenous 5’ and 3’ UTRs for the nucleic acid of interest. Alternatively, UTR sequences that are not endogenous to the nucleic acid of interest can be added by incorporating the UTR sequences into the forward 78ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION and reverse primers or by any other modifications of the template. The use of UTR sequences that are not endogenous to the nucleic acid of interest can be useful for modifying the stability and / or translation efficiency of the RNA For example, it is known that AU-rich elements in 3’ UTR sequences can decrease the stability of mRNA Therefore, 3’ UTRs can be selected or designed to increase the stability of the transcribed RNA based on properties ofUTRs that are well known in the art.

[0249] In some embodiments, the 5’ UTR can contain the Kozak sequence of the endogenous nucleic acid. Alternatively, when a 5’ UTR that is not endogenous to the nucleic acid of interest is being added by PCR as described above, a consensus Kozak sequence can be redesigned by adding the 5’ UTR sequence. Kozak sequences can increase the efficiency of translation of some RNA transcripts, but does not appear to be required for all RNAs to enable efficient translation. The requirement for Kozak sequences for many mRNAs is known in the art. In some embodiments, the 5’ UTR can be 5’UTR of an RNA virus whose RNA genome is stable in cells. In some embodiments, various nucleotide analogues can be used in the 3’ or 5’ UTR to impede exonuclease degradation of the mRNA.

[0250] To enable synthesis of RNA from a DNA template without the need for gene cloning, a promoter of transcription should be attached to the DNA template upstream of the sequence to be transcribed. When a sequence that functions as a promoter for an RNA polymerase is added to the 5’ end of the forward primer, the RNA polymerase promoter becomes incorporated into the PCR product upstream of the open reading frame that is to be transcribed. In one preferred embodiment, the promoter is a T7 polymerase promoter, as described elsewhere herein. Other useful promoters include, but are not limited to, T3 and SP6 RNA polymerase promoters.

[0251] Consensus nucleic acid sequences for T7, T3 and SP6 promoters are known in the art.

[0252] In an embodiment, the mRNA has both a cap on the 5’ end and a 3’ poly(A) tail which determine ribosome binding, initiation of translation and stability mRNA in the cell. On a circular DNA template, for instance, plasmid DNA, RNA polymerase produces a long concatameric product which is not suitable for expression in eukaryotic cells. The transcription of plasmid DNA linearized at the end of the 3’ UTR results in normal sized mRNA which is not effective in eukaryotic transfection even if it is polyadenylated after transcription.

[0253] On a linear DNA template, phage T7 RNA polymerase can extend the 3’ end of the transcript beyond the last base of the template (Schenborn and Mierendorf, Nuc Acids Res., 13:6223-36 (1985); Nacheva and Berzal-Herranz, Eur. J. Biochem., 270:1485-65 (2003). 79ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0254] The conventional method of integration of polyA / T stretches into a DNA template is molecular cloning. However, polyA / T sequence integrated into plasmid DNA can cause plasmid instability, which is why plasmid DNA templates obtained from bacterial cells are often highly contaminated with deletions and other aberrations. This makes cloning procedures not only laborious and time consuming but often not reliable. That is why a method which allows construction of DNA templates with polyA / T 3’ stretch without cloning highly desirable.

[0255] The polyA / T segment of the transcriptional DNA template can be produced during PCR by using a reverse primer containing a polyT tail, such as 100T tail (SEQ ID NO: 257) (size can be 50-5000 T (SEQ ID NO: 258)), or after PCR by any other method, including, but not limited to, DNA ligation or in vitro recombination. Poly(A) tails also provide stability to RNAs and reduce their degradation. Generally, the length of a poly(A) tail positively correlates with the stability of the transcribed. RNA In some embodiments, the poly(A) tail is between 100 and 5000 adenosines (SEQ ID NO: 259).

[0256] Poly(A) tails of RNAs can be further extended following in vitro transcription with the use of a poly(A) polymerase, such as E. coli polyA polymerase (E-PAP). In some embodiments, increasing the length of a poly(A) tail from 100 nucleotides to between 300 and 400 nucleotides (SEQ ID NO: 260) results in about a two-fold increase in the translation efficiency of the RNA Additionally, the attachment of different chemical groups to the 3’ end can increase mRNA stability. Such attachment can contain modified / artificial nucleotides, aptamers and other compounds. For example, ATP analogs can be incorporated into the poly(A) tail using poly(A) polymerase. ATP analogs can further increase the stability of the RNA 5’ caps on also provide stability to RNA molecules. In a preferred embodiment, RNAs produced by the methods disclosed herein include a 5’ cap. The 5’ cap is provided using techniques known in the art and described herein (Cougot, et al., Trends in Biochem. Sci., 29:436-444 (2001); Stepinski, et al., RNA, 7:1468-95 (2001); Elango, et al., Biochim. Biophys. Res. Commun., 330:958-966 (2005)).

[0257] The RNAs produced by the methods disclosed herein can also contain an internal ribosome entry site (IRES) sequence. The IRES sequence may be any viral, chromosomal or artificially designed sequence which initiates cap-independent ribosome binding to mRNA and facilitates the initiation of translation. Any solutes suitable for cell electroporation, which can contain factors facilitating cellular permeability and viability such as sugars, peptides, lipids, proteins, antioxidants, and surfactants can be included. 80ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0258] RNA can be introduced into target cells using any of a number of different methods, for instance, commercially available methods which include, but are not limited to, electroporation (Amaxa Nucleofector-11 (Amaxa Biosystems, Cologne, Germany)), (ECM 830 (BTX) (Harvard Instruments, Boston, Mass.) or the Gene Pulser II (BioRad, Denver, Colo.), Multiporator (Eppendorf, Hamburg Germany), cationic liposome mediated transfection using lipofection, polymer encapsulation, peptide mediated transfection, or biolistic particle delivery systems such as “gene guns” (see, for example, Nishikawa, et al. Hum Gene Ther., 12(8):861-70 (2001). Non-viral delivery methods

[0259] In some embodiments, non-viral methods can be used to deliver a nucleic acid encoding a CAR described herein into a cell or tissue or a subject. In some embodiments, the non-viral method includes the use of a transposon (also called a transposable element). In some embodiments, a transposon is a piece of DNA that can insert itself at a location in a genome, for example, a piece of DNA that is capable of self-replicating and inserting its copy into a genome, or a piece of DNA that can be spliced out of a longer nucleic acid and inserted into another place in a genome. For example, a transposon comprises a DNA sequence made up of inverted repeats flanking genes for transposition.

[0260] Exemplary methods of nucleic acid delivery using a transposon include a Sleeping

[0261] Beauty transposon system (SBTS) and a piggyBac (PB) transposon system. See, e.g., Aronovich et al. Hum. Mol. Genet. 20.Rl(201 l):Rl4-20; Singh et al. Cancer Res. 15(2008):2961-2971; Huang et al. Mol. Ther.16(2008): 580-589; Grabundzija et al. Mol. Ther. 18(2010): 1200-1209; Kebriaei et al. Blood.122.21(2013):166; Williams. Molecular Therapy 16.9(2008):1515-16; Bell et al. Nat. Protoc. 2.12(2007):3153-65; and Ding et al. Cell. 122.3(2005):473-83, all of which are incorporated herein by reference.

[0262] The SBTS includes two components: 1) a transposon containing a transgene and 2) a source of transposase enzyme. The transposase can transpose the transposon from a carrier plasmid (or other donor DNA) to a target DNA, such as a host cell chromosome / genome. For example, the transposase binds to the carrier plasmid / donor DNA, cuts the transposon (including transgene(s)) out of the plasmid, and inserts it into the genome of the host cell. See, e.g., Aronovich et al.

[0263] Exemplary transposons include a pT2-based transposon. See, e.g., Grabundzija et al. Nucleic Acids Res. 41.3(2013): 1829-47; and Singh et al. Cancer Res. 68.8(2008): 2961- 2971, all of which are incorporated herein by reference. Exemplary transposases include a Tcl / mariner-type transposase, e.g., the SBl0 transposase or the SBl1 transposase (a hyperactive 81ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION transposase which can be expressed, e.g., from a cytomegalovirus promoter). See, e.g., Aronovich et al.; Kebriaei et al.; and Grabundzija et al., all of which are incorporated herein by reference.

[0264] Use of the SBTS permits efficient integration and expression of a transgene, e.g., a nucleic acid encoding a CAR described herein. Provided herein are methods of generating a cell, e.g., T cell or NK cell, that stably expresses a CAR described herein, e.g., using a transposon system such as SBTS. In accordance with methods described herein, in some embodiments, one or more nucleic acids, e.g., plasmids, containing the SBTS components are delivered to a cell (e.g., Tor NK cell). For example, the nucleic acid(s) are delivered by standard methods of nucleic acid (e.g., plasmid DNA) delivery, e.g., methods described herein, e.g., electroporation, transfection, or lipofection. In some embodiments, the nucleic acid contains a transposon comprising a transgene, e.g., a nucleic acid encoding a CAR described herein. In some embodiments, the nucleic acid contains a transposon comprising a transgene (e.g., a nucleic acid encoding a CAR described herein) as well as a nucleic acid sequence encoding a transposase enzyme. In some embodiments, a system with two nucleic acids is provided, e.g., a dual-plasmid system, e.g., where a first plasmid contains a transposon comprising a transgene, and a second plasmid contains a nucleic acid sequence encoding a transposase enzyme. For example, the first and the second nucleic acids are co-delivered into a host cell.

[0265] In some embodiments, cells, e.g., Tor NK cells, are generated that express a CAR described herein by using a combination of gene insertion using the SBTS and genetic editing using a nuclease (e.g., Zinc finger nucleases (ZFNs), Transcription Activator-Like Effector Nucleases (TALENs), the CRISPR / Cas system, or engineered meganuclease re- engineered homing endonucleases ). In some embodiments, use of a non-viral method of delivery permits reprogramming of cells, e.g., Tor NK cells, and direct infusion of the cells into a subject. Advantages of non-viral vectors include but are not limited to the ease and relatively low cost of producing sufficient amounts required to meet a patient population, stability during storage, and lack of immunogenicity. Nucleic Acid Constructs Encoding a CAR

[0266] The present disclosure also provides nucleic acid molecules encoding one or more CAR constructs described herein. In some embodiments, the nucleic acid molecule is provided as a messenger RNA transcript. In some embodiments, the nucleic acid molecule is provided as a DNA construct. 82ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0267] The nucleic acid sequences coding for the desired molecules can be obtained using recombinant methods known in the art, such as, for example by screening libraries from cells expressing the gene, by deriving the gene from a vector known to include the same, or by isolating directly from cells and tissues containing the same, using standard techniques. Alternatively, the gene of interest can be produced synthetically, rather than cloned.

[0268] The present disclosure also provides vectors in which a DNA of the present disclosure is inserted. Vectors derived from retroviruses such as the lentivirus are suitable tools to achieve long-term gene transfer since they allow long-term, stable integration of a transgene and its propagation in daughter cells. Lentiviral vectors have the added advantage over vectors derived from onco-retroviruses such as murine leukemia viruses in that they can transduce non- proliferating cells, such as hepatocytes. They also have the added advantage of low immunogenicity.

[0269] In some embodiments, the vector comprising the nucleic acid encoding the desired CAR of the invention is an adenoviral vector (A5 / 35). In some embodiments, the expression of nucleic acids encoding CARs can be accomplished using of transposons such as sleeping beauty, crisper, CAS9, and zinc finger nucleases. See below June et al. 2009Nature Reviews Immunology 9.10: 704-716, which is incorporated herein by reference.

[0270] In brief summary, the expression of natural or synthetic nucleic acids encoding CARs is typically achieved by operably linking a nucleic acid encoding the CAR polypeptide or portions thereof to a promoter, and incorporating the construct into an expression vector. The vectors can be suitable for replication and integration eukaryotes. Typical cloning vectors contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence.

[0271] The expression constructs of the present disclosure may also be used for nucleic acid immunization and gene therapy, using standard gene delivery protocols. Methods for gene delivery are known in the art. See, e.g., U.S. Pat. Nos. 5,399,346, 5,580,859, 5,589,466, incorporated by reference herein in their entireties. In some embodiments, the disclosure provides a gene therapy vector.

[0272] The nucleic acid can be cloned into a number of types of vectors. For example, the nucleic acid can be cloned into a vector including, but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.

[0273] Further, the expression vector may be provided to a cell in the form of a viral vector Viral vector technology is well known in the art and is described, for example, in Sambrook et 83ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION al., 2012, MOLECULAR CLONING: A LABORATORY MANUAL, volumes 1 -4, Cold Spring Harbor Press, NY), and in other virology and molecular biology manuals. Viruses, which are useful as vectors include, but are not limited to, retroviruses, adenoviruses, adeno- associated viruses, herpes viruses, and lentiviruses. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers, (e.g., WO 01 / 96584; WO 01 / 29058; and U.S. Pat. No. 6,326,193).

[0274] A number of viral based systems have been developed for gene transfer into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. A selected gene can be inserted into a vector and packaged in retroviral particles using techniques known in the art. The recombinant virus can then be isolated and delivered to cells of the subject either in vivo or ex vivo. A number of retroviral systems are known in the art. In some embodiments, adenovirus vectors are used. A number of adenovirus vectors are known in the art. In some embodiments, lentivirus vectors are used.

[0275] Additional promoter elements, e.g., enhancers, regulate the frequency of transcriptional initiation. Typically, these are located in the region 30-110 bp upstream of the start site, although a number of promoters have been shown to contain functional elements downstream of the start site as well. The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or moved relative to one another. In the thymidine kinase (tk) promoter, the spacing between promoter elements can be increased to 50 bp apart before activity begins to decline. Depending on the promoter, it appears that individual elements can function either cooperatively or independently to activate transcription. Exemplary promoters include the CMV IE gene, EF-la, ubiquitin C, or phosphoglycerokinase (PGK) promoters.

[0276] An example of a promoter that is capable of expressing a CAR transgene in a mammalian T cell is the EF-1 alpha (EFla) promoter. The native EFla promoter drives expression of the alpha subunit of the elongation factor-1 complex, which is responsible for the enzymatic delivery of aminoacyl tRNAs to the ribosome. The EFla promoter has been extensively used in mammalian expression plasmids and has been shown to be effective in driving CAR expression from transgenes cloned into a lentiviral vector. See, e.g., Milone et al., Mol. Ther.17(8): 1453- 1464 (2009). In some embodiments, the EFla promoter comprises the sequence as known in the art.

[0277] Another example of a promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence capable 84ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION of driving high levels of expression of any polynucleotide sequence operatively linked thereto. However, other constitutive promoter sequences may also be used, including, but not limited to the simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, an avian leukemia virus promoter, an Epstein-Barr virus immediate early promoter, a Rous sarcoma virus promoter, as well as human gene promoters such as, but not limited to, the actin promoter, the myosin promoter, the elongation factor-lex promoter, the hemoglobin promoter, and the creatine kinase promoter. Further, the invention should not be limited to the use of constitutive promoters. Inducible promoters are also contemplated as part of the invention. The use of an inducible promoter provides a molecular switch capable of turning on expression of the polynucleotide sequence which it is operatively linked when such expression is desired, or turning off the expression when expression is not desired. Examples of inducible promoters include, but are not limited to a metallothionine promoter, a glucocorticoid promoter, a progesterone promoter, and a tetracycline promoter.

[0278] In order to assess the expression of a CAR polypeptide or portions thereof, the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of expressing cells from the population of cells sought to be transfected or infected through viral vectors. In other embodiments, the selectable marker may be carried on a separate piece of DNA and used in a co- transfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers include, for example, antibiotic-resistance genes, such as neo and the like.

[0279] Reporter genes are used for identifying potentially transfected cells and for evaluating the functionality of regulatory sequences. In general, a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and that encodes a polypeptide whose expression is manifested by some easily detectable property, e.g., enzymatic activity. Expression of the reporter gene is assayed at a suitable time after the DNA has been introduced into the recipient cells. Suitable reporter genes may include genes encoding luciferase, beta- galactosidase, chloramphenicol acetyl transferase, secreted alkaline phosphatase, or the green fluorescent protein gene (e.g., Ui-Tei et al., 2000 FEBS Letters 479: 79-82). Suitable expression systems are well known and may be prepared using known techniques or obtained commercially. In general, the construct with the minimal 5’ flanking region showing the highest level of expression of reporter gene is identified as the promoter. Such promoter regions 85ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION may be linked to a reporter gene and used to evaluate agents for the ability to modulate promoter- driven transcription.

[0280] Methods of introducing and expressing genes into a cell are known in the art. In the context of an expression vector, the vector can be readily introduced into a host cell, e.g., mammalian, bacterial, yeast, or insect cell by any method in the art. For example, the expression vector can be transferred into a host cell by physical, chemical, or biological means.

[0281] Physical methods for introducing a polynucleotide into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. Methods for producing cells comprising vectors and / or exogenous nucleic acids are well-known in the art. See, for example, Sambrook et al., 2012, MOLECULAR CLONING: A LABORATORY MANUAL, volumes 1 -4, Cold Spring Harbor Press, NY). A preferred method for the introduction of a polynucleotide into a host cell is calcium phosphate transfection.

[0282] Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells. Other viral vectors can be derived from lentivirus, poxviruses, herpes simplex virus 1, adenoviruses and adeno-associated viruses, and the like. See, for example, U.S. Pat. Nos. 5,350,674 and 5,585,362.

[0283] Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as a delivery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle). Other methods of state-of-the-art targeted delivery of nucleic acids are available, such as delivery of polynucleotides with targeted nanoparticles or other suitable sub-micron sized delivery system.

[0284] In the case where a non-viral delivery system is utilized, an exemplary delivery vehicle is a liposome. The use of lipid formulations is contemplated for the introduction of the nucleic acids into a host cell (in vitro, ex vivo or in vivo). In some embodiments, the nucleic acid may be associated with a lipid. The nucleic acid associated with a lipid may be encapsulated in the aqueous interior of a liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via a linking molecule that is associated with both the liposome and the oligonucleotide, entrapped in a liposome, complexed with a liposome, dispersed in a solution containing a lipid, mixed with a lipid, combined with a lipid, contained 86ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION as a suspension in a lipid, contained or complexed with a micelle, or otherwise associated with a lipid. Lipid, lipid / DNA or lipid / expression vector associated compositions are not limited to any particular structure in solution. For example, they may be present in a bilayer structure, as micelles, or with a “collapsed” structure. They may also simply be interspersed in a solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty substances which may be naturally occurring or synthetic lipids. For example, lipids include the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds which contain long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes.

[0285] Lipids suitable for use can be obtained from commercial sources. For example, dimyristyl phosphatidylcholine (“DMPC”) can be obtained from Sigma, St. Louis, MO; dicetyl phosphate (“DCP”) can be obtained from K & K Laboratories (Plainview, NY); cholesterol (“Choi”) can be obtained from Calbiochem-Behring; dimyristyl phosphatidylglycerol (“DMPG”) and other lipids may be obtained from Avanti Polar Lipids, Inc. (Birmingham, AL.). Stock solutions of lipids in chloroform or chloroform / methanol can be stored at about -20°C. Chloroform is used as the only solvent since it is more readily evaporated than methanol. “Liposome” is a generic term encompassing a variety of single and multilamellar lipid vehicles formed by the generation of enclosed lipid bilayers or aggregates. Liposomes can be characterized as having vesicular structures with a phospholipid bilayer membrane and an inner aqueous medium. Multilamellar liposomes have multiple lipid layers separated by aqueous medium. They form spontaneously when phospholipids are suspended in an excess of aqueous solution. The lipid components undergo self-rearrangement before the formation of closed structures and entrap water and dissolved solutes between the lipid bilayers (Ghosh et al., 1991 Glycobiology 5: 505-10).

[0286] However, compositions that have different structures in solution than the normal vesicular structure are also encompassed. For example, the lipids may assume a micellar structure or merely exist as nonuniform aggregates of lipid molecules. Also contemplated are lipofectamine- nucleic acid complexes.

[0287] Regardless of the method used to introduce exogenous nucleic acids into a host cell or otherwise expose a cell to the inhibitor of the present disclosure, in order to confirm the presence of the recombinant DNA sequence in the host cell, a variety of assays may be performed. Such assays include, for example, “molecular biological” assays well known to those of skill in the art, such as Southern and Northern blotting, RT-PCR and PCR; “biochemical” assays, such as detecting the presence or absence of a particular peptide, e.g., 87ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION by immunological means (ELISAs and Western blots) or by assays described herein to identify agents falling within the scope of the invention.

[0288] The present disclosure further provides a vector comprising a CAR encoding nucleic acid molecule. In some embodiments, a CAR vector can be directly transduced into a cell, e.g., a T cell or NK cell. In some embodiments, the vector is a cloning or expression vector, e.g., a vector including, but not limited to, one or more plasmids (e.g., expression plasmids, cloning vectors, minicircles, minivectors, double minute chromosomes), retroviral and lentiviral vector constructs. In some embodiments, the vector is capable of expressing the CAR construct in mammalian T cells or NK cells. In some embodiments, the mammalian T cell is a human T cell. Nucleic Acid Constructs Encoding a Bispecific T Cell Engager

[0289] The present disclosure also provides nucleic acid molecules encoding one or more bispecific T cell engager construct described herein. In some embodiments, the nucleic acid molecule is provided as a messenger RNA transcript. In some embodiments, the nucleic acid molecule is provided as a DNA construct.

[0290] The nucleic acid sequences coding for the desired molecules can be obtained using recombinant methods known in the art, such as, for example by screening libraries from cells expressing the gene, by deriving the gene from a vector known to include the same, or by isolating directly from cells and tissues containing the same, using standard techniques. Alternatively, the gene of interest can be produced synthetically, rather than cloned.

[0291] In brief summary, the expression of natural or synthetic nucleic acids encoding bispecific T cell engagers is, in some embodiments, achieved by operably linking a nucleic acid encoding the bispecific T cell engager polypeptide or portions thereof to a promoter, and incorporating the construct into an expression vector. Typical cloning vectors contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence.

[0292] The nucleic acid can be cloned into a number of types of vectors. For example, the nucleic acid can be cloned into a vector including, but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors. Methods of Manufacture / Production

[0293] The present disclosure also provides methods of making a cell disclosed herein, e.g., methods of engineering a T cell or NK cell to express a nucleic acid molecule encoding 88ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION one or more CAR constructs described herein. In some embodiments, the manufacturing methods disclosed herein are used to manufacture a cell comprising a nucleic acid molecule encoding two

[0294] CARs disclosed herein (e.g., a tandem and / or dual CAR disclosed herein). In some embodiments, the manufacturing methods disclosed herein are used to manufacture a cell comprising a nucleic acid molecule encoding a diabody CAR disclosed herein, e.g., an anti- NYESO-1 / anti-CD3 diabody CAR disclosed herein. In some embodiments, the manufacturing methods disclosed herein are used to manufacture a cell comprising two nucleic acid molecules, each of which encodes a CAR disclosed herein (e.g., one nucleic acid molecule encoding an anti-NYESO-1 CAR and one nucleic acid molecule encoding an anti-CD3 CAR). In some embodiments, provided herein is a population of cells (for example, immune effector cells, for example, T cells or NK cells) made by any of the manufacturing processes described herein.

[0295] In some embodiments, the methods disclosed herein may manufacture immune effector cells engineered to express one or more CARs in less than 24 hours. Without wishing to be bound by theory, the methods provided herein preserve the undifferentiated phenotype of T cells, such as naive T cells, during the manufacturing process. These CAR-expressing cells with an undifferentiated phenotype may persist longer and / or expand better in vivo after infusion. In some embodiments, CART cells produced by the manufacturing methods provided herein comprise a higher percentage of stem cell memory T cells, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using single-cell or bulk RNA-seq or flow cytometry using markers known in the art. In some embodiments, CART cells produced by

[0296] In some embodiments, the manufacturing methods provided herein comprise a lower percentage of effector T cells, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using single-cell RNA-seq. In some embodiments, CART cells produced by the manufacturing methods provided herein better preserve the sternness of T cells, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. In some

[0297] In some embodiments, CART cells produced by the manufacturing methods provided herein show a lower level of hypoxia, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. In some embodiments, CART cells produced by the manufacturing methods provided herein show a lower level of autophagy, compared to CART cells produced by the traditional manufacturing process, e.g., 89ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION as measured using scRNA-seq. In some embodiments, the methods disclosed herein do not involve using a bead, such as Dynabeads® (for example, CD3 / CD28 Dynabeads®), and do not involve a de-beading step. In some embodiments, the CART cells manufactured by the methods disclosed herein may be administered to a subject with minimal ex vivo expansion, for example, less than 2 days, less than 1 day, less than 12 hours, less than 8 hours, less than 6 hours, less than 4 hours, less than 3 hours, less than 2 hours, less than 1 hour, or no ex vivo expansion. Accordingly, the methods described herein provide a fast manufacturing process of making improved CAR-expressing cell products for use in treating a disease in a subject.

[0298] In some embodiments, the present disclosure provides methods of making a population of cells (for example, T cells) that express a chimeric antigen receptor (CAR) (e.g., one or more CARs, e.g., two CARs) comprising: (i) contacting a population of cells (for example, T cells, for example, T cells isolated from a frozen or fresh leukapheresis product) with an agent that stimulates a CD3 / TCR complex and / or an agent that stimulates a costimulatory molecule on the surface of the cells; (ii) contacting the population of cells (for example, T cells) with a nucleic acid molecule(s) (for example, a DNA or RNA molecule) encoding the CAR(s), thereby providing a population of cells (for example, T cells) comprising the nucleic acid molecule, and harvesting the population of cells (for example, T cells) for storage (for example, reformulating the population of cells in cryopreservation media) or administration, wherein: (a) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i), for example, no later than 12, 13, 14, 15, 16, 17, or 18 hours after the beginning of step (i), for example, no later than 18 hours after the beginning of step (i), and step (iii) is performed no later than 26 hours after the beginning of step (i), for example, no later than 22, 23, or 24 hours after the beginning of step (i), for example, no later than 24 hours after the beginning of step (i); (b) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i), for example, no later than 12, 13, 14, 15, 16, 17, or 18 hours after the beginning of step (i), for example, no later than 18 hours after the beginning of step (i), and step (iii) is performed no later than 30 hours after the beginning of step (ii), for example, no later than 22, 23, 24, 25, 26, 27, 28, 29, or 30 hours after the beginning of step (ii); and / or (c) the population of cells from step (iii) are not expanded, or expanded by no more than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%, for example, no more than 10%, as assessed by the number of living cells compared to the population of cells at the beginning of step (i); or d) the population of cells from step (iii) are fewer, or less by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, or 70%, for example, as assessed by the number of living cells compared to the population of cells at the beginning of step (i). In some embodiments, 90ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION the nucleic acid molecule in step (ii) is a DNA molecule. In some embodiments, the nucleic acid molecule in step (ii) is an RNA molecule. In some embodiments, the nucleic acid molecule in step (ii) is on a viral vector, for example, a viral vector chosen from a lentivirus vector, an adenoviral vector, or a retrovirus vector. In some embodiments, the nucleic acid molecule in step (ii) is on a non-viral vector. In some embodiments, the nucleic acid molecule in step (ii) is on a plasmid. In some embodiments, the nucleic acid molecule in step (ii) is not on any vector. In some embodiments, step (ii) comprises transducing the population of cells (for example, T cells) a viral vector(s) comprising a nucleic acid molecule encoding the CAR(s). In some embodiments of the aforementioned methods, the methods further comprise adding an adjuvant or a transduction enhancement reagent in the cell culture medium to enhance transduction efficiency. In some embodiments, the adjuvant or transduction enhancement reagent comprises a cationic polymer. In some embodiments, the adjuvant or transduction enhancement reagent is selected from: LentiBOOS T™ (Sirion Biotech), vectofusin-1, F108 (Poloxamer 338 or Pluronic® F-38), hexadimethrine bromide (Polybrene), PEA, Pluronic F68, Pluronic Fl27(Poloxamer 407 or Synperonic PE / F-127), Protamine Sulfate, Synperonic or LentiTrans™. In some embodiments, the adjuvant is LentiBOOST™ (Sirion Biotech). In other embodiments, the adjuvant is Fl08 (Poloxamer 338 or Pluronic® F-38). In other embodiments, the adjuvant is Pluronic Fl27 (Poloxamer 407 or Synperonic PE / F-127).

[0299] In some embodiments, the population of cells (for example, T cells) is collected from an apheresis sample (for example, a leukapheresis sample) from a subject. In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject and shipped as a frozen sample (for example, a cryopreserved sample) to a cell manufacturing facility. Then the frozen apheresis sample is thawed, and T cells (for example, CD4+ T cells and / or CD8+ T cells) are selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are then seeded for CART manufacturing using the activation process described herein. In some embodiments, the selected T cells (for example, CD4+ T cells and / or CD8+ T cells) undergo one or more rounds of freeze- thaw before being seeded for CART manufacturing.

[0300] In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject and shipped as a fresh product (for example, a product that is not frozen) to a cell manufacturing facility. T cells (for example, CD4+ T cells and / or CD8+ T cells) are selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells 91ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION and / or CD8+ T cells) are then seeded for CART manufacturing using the activation process described herein. In some embodiments, the selected T cells (for example, CD4+ T cells and / or CD8+ T cells) undergo one or more rounds of freeze-thaw before being seeded for CART manufacturing.

[0301] In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject. T cells (for example, CD4+ T cells and / or CD8+ T cells) are selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are then shipped as a frozen sample (for example, a cryopreserved sample) to a cell manufacturing facility. The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are later thawed and seeded for CART manufacturing using the activation process described herein.

[0302] In some embodiments, cells (for example, T cells) are contacted with anti-CD3 and anti- CD28 antibodies and, for example, immediately followed by transduction with a vector (for example, a lentiviral vector) (e.g. one or more vectors) encoding a CAR (e.g. one or more CARs). 24 hours after culture initiation, the cells are washed and formulated for storage or administration.

[0303] Without wishing to be bound by theory, brief CD3 and CD28 stimulation may promote efficient transduction of self-renewing T cells. Compared to traditional CART manufacturing approaches, the activation process provided herein does not involve prolonged ex vivo expansion. Similar to the cytokine process, the activation process provided herein also preserves undifferentiated T cells during CART manufacturing.

[0304] In some embodiments, cells (for example, T cells) are contacted with anti-CD3 and anti- CD28 antibodies for, for example, 12 hours, followed by transduction with a vector (for example, a lentiviral vector) (e.g. one or more vectors) encoding a CAR (e.g. one or more CARs). 24 hours after culture initiation, the cells are washed and formulated for storage or administration.

[0305] Without wishing to be bound by theory, brief CD3 and CD28 stimulation may promote efficient transduction of self-renewing T cells. Compared to traditional CART manufacturing approaches, the activation process provided herein does not involve prolonged ex vivo expansion. Similar to the cytokine process, the activation process provided herein also preserves undifferentiated T cells during CART manufacturing.

[0306] In some embodiments, the population of cells is contacted with an agent that stimulates a CD3 / TCR complex and / or an agent that stimulates a costimulatory molecule on 92ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION the surface of the cells. In some embodiments, T cells are selected using anti-CD4 and anti- CDS beads by positive selection, using, for example, a cell sorting machine (for example, a CliniMACS® Prodigy® device).

[0307] In some embodiments, T cells are selected using anti-CD45RA and anti-CCR7 beads by positive selection, using, for example, a cell sorting machine (for example, a CliniMACS® Prodigy® device).

[0308] In some embodiments, T cells are selected using anti-CD45RA and anti-CD27 beads by positive selection, using, for example, a cell sorting machine (for example, a CliniMACS® Prodigy® device).

[0309] In some embodiments, T cells are selected using anti-CD3 and anti-CD28 beads by positive selection, using, for example, a cell sorting machine (for example, a CliniMACS® Prodigy® device).

[0310] In some embodiments, T cells are selected using anti-lineage beads (except for T cell) by negative selection, using, for example, a cell sorting machine (for example, a CliniMACS® Prodigy® device).

[0311] In some embodiments, the agent that stimulates a CD3 / TCR complex is an agent that stimulates CD3. In some embodiments, the agent that stimulates a costimulatory molecule is an agent that stimulates CD28, ICOS, CD27, HVEM, LIGHT, CD40, 4-lBB, OX40, DR3, GITR, CD30, TIMI, CD2, CD226, or any combination thereof. In some embodiments, the agent that stimulates a costimulatory molecule is an agent that stimulates CD28. In some embodiments, the agent that stimulates a CD3 / TCR complex is chosen from an antibody (for example, a single- domain antibody (for example, a heavy chain variable domain antibody), a peptibody, a Fab fragment, or a scFv), a small molecule, or a ligand (for example, a naturally- existing, recombinant, or chimeric ligand). In some embodiments, the agent that stimulates a CD3 / TCR complex is an antibody. In some embodiments, the agent that stimulates a CD3 / TCR complex is an anti-CD3 antibody. In some embodiments, the agent that stimulates a costimulatory molecule is chosen from an antibody (for example, a single-domain antibody (for example, a heavy chain variable domain antibody), a peptibody, a Fab fragment, or a scFv), a small molecule, or a ligand (for example, a naturally-existing, recombinant, or chimeric ligand). In some embodiments, the agent that stimulates a costimulatory molecule is an antibody. In some embodiments, the agent that stimulates a costimulatory molecule is an anti-CD28 antibody. In some embodiments, the agent that stimulates a CD3 / TCR complex or the agent that stimulates a costimulatory molecule does not comprise a bead. In some embodiments, the agent that stimulates a CD3 / TCR complex comprises an anti-CD3 antibody 93ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION covalently attached to a colloidal polymeric nanomatrix. In some embodiments, the agent that stimulates a costimulatory molecule comprises an anti-CD28 antibody covalently attached to a colloidal polymeric nanomatrix. In some embodiments, the agent that stimulates a CD3 / TCR complex and the agent that stimulates a costimulatory molecule comprising T Cell TransAct™.

[0312] In some embodiments, the matrix comprises or consists of a polymeric, for example, biodegradable or biocompatible inert material, for example, which is non-toxic to cells. In some embodiments, the matrix is composed of hydrophilic polymer chains, which obtain maximal mobility in aqueous solution due to hydration of the chains. In some embodiments, the mobile matrix may be of collagen, purified proteins, purified peptides, polysaccharides, glycosaminoglycans, or extracellular matrix compositions. A polysaccharide may include for example, cellulose ethers, starch, gum arabic, agarose, dextran, chitosan, hyaluronic acid, pectins, xanthan, guar gum or alginate. Other polymers may include polyesters, polyethers, polyacrylates, polyacrylamides, polyamines, polyethylene imines, polyquaternium polymers, polyphosphazenes, polyvinylalcohols, polyvinylacetates, polyvinylpyrrolidones, block copolymers, or polyurethanes. In some embodiments, the mobile matrix is a polymer of dextran.

[0313] In some embodiments, the population of cells is contacted with a nucleic acid molecule (e.g. one or more nucleic acid molecules) encoding a CAR (e.g. one or more CARs). In some embodiments, the population of cells is transduced with a DNA molecule (e.g. one or more DNA ) encoding a CAR (e.g. one or more CARs).

[0314] In some embodiments, in the case of a co-transduction of two nucleic acid molecules (e.g., lentiviral vectors), each of which encodes a CAR disclosed herein (e.g., one nucleic acid molecule encoding an anti-CD22 CAR and one nucleic acid molecule encoding an anti-CD 19 CAR, as disclosed herein), each of the vectors containing nucleic acid molecules encoding the

[0315] CAR can be added to the reaction mixture (e.g., containing a cell population), e.g., at 1:1 ratio.

[0316] Without wishing to be bound by theory, it is believed that, in some embodiments, using different MOIs for the vectors containing nucleic acid molecules that encode distinct CAR molecules may affect the final composition of the cellular population.

[0317] In some embodiments, in the case of a co-transduction of a lentiviral vector encoding an anti- EBV-A02, SL9-A02, Nef_A24, or SARS-B08 and a lentiviral vector encoding an anti- EBV-A02, SL9-A02, Nef_A24, or SARS-B08, a population of cells is 94ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION contacted with the first viral vector at an MOI that is higher than, equal to, or less than an MOI at which the population of cells is contacted with the second viral vector. In some embodiments, the population of cells is contacted with the first viral vector at an MOI that is higher than an MOI at which the population of cells is contacted with the second viral vector.

[0318] In some embodiments, the population of cells is contacted with the first viral vector at first MOI and with the second viral vector at a second MOI, such that a resultant population of cells comprises a first population of cells that comprise the first CAR but not the second CAR, a second population of cells that comprise the second car, but not the first CAR, and a third population of cells that comprise both the first CAR and the second CAR. Co-transduction methods are as described in the art, e.g., in WO 2021 / 108661 which is incorporated by reference in its entirety.

[0319] In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs simultaneously with contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0.5 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 20 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 19 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 18 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 17 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or 95ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 16 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 15 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 14 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 14 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 13 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 12 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 11 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 10 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 9 hours after the beginning of contacting the population of cells with the agent that stimulates 96ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 8 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 7 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 6 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 5 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 4 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 3 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs no later than 2 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting thepopulation of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 1 hour after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule(s) encoding the CAR(s) occurs no later than 30 minutes after the beginning of contacting the population of cells with 97ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above.

[0320] In some embodiments, the population of cells is harvested for storage or administration. In some embodiments, the population of cells is harvested for storage or administration no later than 72, 60, 48, 36, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, or 18 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 26 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 25 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 24 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 23 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 22 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above.

[0321] In some embodiments, the population of cells is not expanded ex vivo.

[0322] In some embodiments, the population of cells is expanded by no more than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, or 60%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 5%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule 98ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 10%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 15%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 20%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 25%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 30%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 35%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 40%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule on the surface of the cells described above.

[0323] In some embodiments, the population of cells is expanded by no more than 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 11, 12, 16, 20, 24, 36, or 48 hours, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above.

[0324] In some embodiments, the activation process is conducted in serum free cell media. In some embodiments, the activation process is conducted in cell media comprising one or 99ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION more cytokines chosen from: IL-2, IL-15 (for example, hetIL-15 (IL-15 / sIL-l 5Ra)), or IL-6 (for example, IL-6 / sIL-6Ra). In some embodiments, hetIL-15 comprises the amino acid sequence of NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVT AMKCFLLELQVISLESGDASIH DTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSITCPPP MS VEHADIWVKSYSL YSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDP ALVHQRPAPPSTVTTAGVTPQPESLSPSGKEPAASSPSSNNTAATTAAIVPGSQLMPSKS P STGTTEISSHESSHGTPSQTTAKNWELTASASHQPPGVYPQG(SEQ ID NO: 253).

[0325] In some embodiments, hetIL-15 comprises an amino acid sequence having at least about 70, 75, 80, 85,90, 95, or 99% identity to SEQ ID NO: 309. In some embodiments, the activation process is conducted in cell media comprising a LSD 1 inhibitor. In some embodiments, the activation process is conducted in cell media comprising a MALTI inhibitor. In some embodiments, the serum free cell media comprises a serum replacement. In some embodiments, the serum replacement is CTS™ Immune Cell Serum Replacement (ICSR). In some embodiments, the level of ICSR can be, for example, up to 5%, for example, about 1%, 2%, 3%, 4%, or 5%. Without wishing to be bound by theory, using cell media, for example, Rapid Media, comprising ICSR, for example, 2% ICSR may improve cell viability during a manufacture process described herein.

[0326] In some embodiments, the present disclosure provides methods of making a population of cells (for example, T cells) that express a chimeric antigen receptor (CAR) comprising: (a) providing an apheresis sample (for example, a fresh or cryopreserved leukapheresis sample) collected from a subject; (b) selecting T cells from the apheresis sample (for example, using negative selection, positive selection, or selection without beads); (c)seeding isolated T cells at, for example, 1 x 106to 1 x 107cells / mL; (d) contacting T cells withan agent that stimulates T cells, for example, an agent that stimulates a CD3 / TCR complex and / or an agent that stimulates a costimulatory molecule on the surface of the cells (for example, contacting T cells with anti-CD3 and / or anti-CD28 antibody, for example, contacting T cells with TransAct); (e) contacting T cells with a nucleic acid molecule(s) (for example, a DNA or RNA molecule) encoding the CAR(s) (for example, contacting T cells with a virus comprising a nucleic acid molecule(s) encoding the CAR(s)) for, for example, 6-48 hours, for example, 20-28 hours; and (f) washing and harvesting T cells for storage (for example, reformulating T cells in cryopreservation media) or administration. In some embodiments, step 100ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION (f) is performed no later than 30 hours after the beginning of step (d) or (e), for example, no later than 22, 23, 24, 25, 26, 27, 28, 29, or 30 hours after the beginning of step (d) or (e).

[0327] In some embodiments, provided herein is a population of cells (for example, immune effector cells, for example, T cells or NK cells) made by any of the manufacturing processes described herein (e.g., the Activation Process described herein).

[0328] In some embodiments, the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells at the end of the manufacturing process (for example, at the end of the cytokine process or the activation process described herein) (1) is the same as, (2) differs, for example, by no more than 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15%, from, or (3) is increased, for example, by at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25%, as compared to, the percentage of naïve cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the population of cells at the beginning of the manufacturing process (for example, at the beginning of the cytokine process or the activation process described herein). In some embodiments, the population of cells at the end of the manufacturing process (for example, at the end of the cytokine process or the activation process described herein) shows a higher percentage of naïve cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells (for example, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50% higher), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0329] In some embodiments, the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells at the end of the manufacturing process (for example, at the end of the cytokine process or the activation process described herein) is not less than 20, 25, 30, 35, 40, 45, 50, 55, or 60%.

[0330] In some embodiments, the percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells, CD45RO+ central memory T cells, and / or CCR7+ central memory T cells, in the population of cells at the end of the manufacturing process (for example, at the end of the cytokine process or the activation process described herein) (1) is the same as, (2) differs, for example, by no more than 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15% from, or (3) is decreased, for example, by at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25%, as compared to, the percentage of 101ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells, in the population of cells at the beginning of the manufacturing process (for example, at the beginning of the cytokine process or the activation process described herein). In some embodiments, the population of cells at the end of the manufacturing process (for example, at the end of the cytokine process or the activation process described herein) shows a lower percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells (for example, at least 5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,25,30,35,40, 45, or 50% lower), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5,6,7,8,9,10,11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3,4,5,6,7,8,9,10,11,12,13,14, or15 days).

[0331] In some embodiments, the percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells, in the population of cells at the end of the manufacturing process (for example, at the end of the cytokine process or the activation process described herein) is no more than 40, 45, 50, 55, 60, 65, 70, 75, or 80%.

[0332] In some embodiments, the population of cells at the end of the manufacturing process (for example, at the end of the cytokine process or the activation process described herein) after being administered in vivo, persists longer or expands at a higher level (for example, at least 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90% higher), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0333] In some embodiments, the population of cells has been enriched for IL6R- expressing cells (for example, cells that are positive for IL6Ra and / or IL6R ) prior to the beginning of the manufacturing process (for example, prior to the beginning of the cytokine process or the activation process described herein). In some embodiments, the population of cells comprises, for example, no less than 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80% ofIL6R- expressing cells (for example, cells that are positive for IL6Ra and / or IL6R ) at the beginning of the manufacturing process (for example, at the beginning of the cytokine process or the activation process described herein). Cytokine Process 102ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0334] In some embodiments, the present disclosure provides methods of making a population of cells (for example, T cells) that express a chimeric antigen receptor (CAR) (e.g., one or more CARs, e.g., two CARs) comprising: (1) contacting a population of cells with a cytokine chosen from IL-2, IL-7, IL-15, IL-21, IL-6, or a combination thereof, (2) contacting the population of cells (for example, T cells) with a nucleic acid molecule(s) (for example, a DNA or RNA molecule) encoding the CAR(s), thereby providing a population of cells (for example, T cells) comprising the nucleic acid molecule, and (3) harvesting the population of cells (for example, T cells) for storage (for example, reformulating the population of cells in cryopreservation media) or administration, wherein: (a) step (2) is performed together with step (1) or no later than 5 hours after the beginning of step (1), for example, no later than 1, 2, 3, 4, or 5 hours after the beginning of step (1), and step (3) is performed no later than 26 hours after the beginning of step (1), for example, no later than 22, 23, or 24 hours after the beginning of step (1), for example, no later than 24 hours after the beginning of step (1), or (b) the population of cells from step (3) are not expanded, or expanded by no more than 5, 10, 15, 20, 25, 30, 35, or 40%, for example, no more than 10%, for example, as assessed by the number of living cells, compared to the population of cells at the beginning of step (1). In some embodiments, the nucleic acid molecule in step (2) is a DNA molecule. In some embodiments, the nucleic acid molecule in step (2) is an RNA molecule. In some embodiments, the nucleic acid molecule in step (2) is on a viral vector, for example, a viral vector chosen from a lentivirus vector, an adenoviral vector, or a retrovirus vector. In some embodiments, the nucleic acid molecule in step (2) is on a non-viral vector. In some embodiments, the nucleic acid molecule in step (2) is on a plasmid. In some embodiments, the nucleic acid molecule in step (2) is not on any vector. In some embodiments, step (2) comprises transducing the population of cells (for example, T cells) with a viral vector comprising a nucleic acid molecule(s) encoding the CAR(s). In some embodiments, the cells are engineered to comprise a nucleic acid molecule encoding a tandem or dual CAR disclosed herein In some embodiments, the cells are engineered to comprise a nucleic acid molecule encoding a diabody CAR disclosed herein

[0335] In some embodiments, the cells are engineered to comprise two nucleic acid molecules, each of which encodes a CAR disclosed herein. In some embodiments, the population of cells (for example, T cells) is collected from an apheresis sample (for example, a leukapheresis sample) from a subject. In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject and shipped as a frozen sample (for example, a cryopreserved sample) to a cell manufacturing facility. The frozen apheresis sample is then thawed, and T cells (for example, CD4+ T cells and / or CD8+ T cells) are 103ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are then seeded for CART manufacturing using the cytokine process described herein. In some embodiments, at the end of the cytokine process, the CART cells are cryopreserved and later thawed and administered to the subject. In some embodiments, the selected T cells (for example, CD4+ T cells and / or CD8+ T cells) undergo one or more rounds of freeze-thaw before being seeded for CART manufacturing.

[0336] In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject and shipped as a fresh product (for example, a product that is not frozen) to a cell manufacturing facility. T cells (for example, CD4+ T cells and / or CD8+ T cells) are selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are then seeded for CART manufacturing using the cytokine process described herein. In some embodiments, the selected T cells (for example, CD4+ T cells and / or CD8+ T cells) undergo one or more rounds of freeze-thaw before being seeded for CART manufacturing.

[0337] In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject. T cells (for example, CD4+ T cells and / or CD8+ T cells) are selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are then shipped as a frozen sample (for example, a cryopreserved sample) to a cell manufacturing facility. The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are later thawed and seeded for CART manufacturing using the cytokine process described herein.

[0338] In some embodiments, after cells (for example, T cells) are seeded, one or more cytokines (for example, one or more cytokines chosen from IL-2, IL-7, IL-15 (for example, hetIL-15 (IL15 / sIL-15Ra)), IL-21, or IL-6 (for example, IL-6 / sIL-6R)) as well as a vector (for example, a lentiviral vector) (e.g. one or more vectors) encoding a CAR (e.g., one or more CARs) are added to the cells. After incubation for 20-24 hours, the cells are washed and formulated for storage or administration.

[0339] Different from traditional CART manufacturing approaches, the cytokine process provided herein does not involve CD3 and / or CD28 stimulation, or ex vivo T cell expansion. T cells that are contacted with anti-CD3 and anti-CD28 antibodies and expanded extensively ex vivo tend to show differentiation towards a central memory phenotype. Without wishing to 104ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION be bound by theory, the cytokine process provided herein preserves or increases the undifferentiated phenotype of T cells during CART manufacturing, generating a CART product that may persist longer after being infused into a subject.

[0340] In some embodiments, the population of cells is contacted with one or more cytokines (for example, one or more cytokines chosen from IL-2, IL-7, IL-15 (for example, hetIL-15 (IL15 / sIL-15Ra)), IL-21, or IL-6 (for example, IL-6 / sIL-6Ra).

[0341] In some embodiments, the population of cells is contacted with IL-2. In some embodiments, the population of cells is contacted with IL-7. In some embodiments, the population of cells is contacted with IL-15 (for example, hetIL-(ILI 5 / sIL-l 5Ra)). In some embodiments, the population of cells is contacted with IL-21. In some embodiments, the population of cells is contacted with IL-6 (for example, IL-6 / sIL-6Ra). In some embodiments, the population of cells is contacted with IL-2 and IL-7. In some embodiments, the population of cells is contacted with IL-2 and IL-15 (for example, hetIL-15 (IL15 / sIL-15Ra)). In some embodiments, the population of cells is contacted with IL-2 and IL-21. In some embodiments, the population of cells is contacted with IL-2 and IL-6 (for example, IL-6 / sIL-6Ra). In some embodiments, the population of cells is contacted with IL-7 and IL-15 (for example, hetIL-15 (ILI 5 / sIL-l 5Ra)). In some embodiments, the population of cells is contacted with IL-7 and IL-6. In some embodiments, the population of cells is contacted with IL-7 and IL-6 (for example, IL-6 / sIL-6Ra). In some embodiments, the population of cells is contacted with IL- 15 (for example, hetIL-15 (ILl 5 / sIL-l 5Ra)) and IL-21. In some embodiments, the population of cells is contacted with IL-15 (for example, hetIL-15 (IL15 / sIL-15Ra)) and IL-6 (for example, IL-6 / sIL- 6Ra). In some embodiments, the population of cells is contacted with IL- 21 and IL-6 (for example, IL-6 / sIL-6Ra). In some embodiments, the population of cells is contacted with IL-7, IL-15 (for example, hetIL-15 (IL15 / sIL-15Ra)), and IL-21. In some embodiments, the population of cells is further contacted with a LSD1 inhibitor. In some embodiments, the population of cells is further contacted with a MALTl inhibitor.

[0342] In some embodiments, the population of cells is contacted with 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, or 300 U / ml of IL-2. In some embodiments, the population of cells is contacted with 1,2,3,4,5,6, 7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20ng / ml of IL-7. In some embodiments, the population of cells is contacted with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, or 20 ng / ml of IL-15.

[0343] In some embodiments, the population of cells is contacted with a nucleic acid molecule (e.g. one or more nucleic acid molecules) encoding a CAR (e.g., one or more CARs). 105ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION In some embodiments, the population of cells is transduced with a DNA molecule (e.g. one or more DNA molecules) encoding a CAR (e.g. one or more CARs).

[0344] In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs simultaneously with contacting the population of cells with the one or more cytokines described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs no later than 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs no later than 5 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs no later than 4 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs no later than 3 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs no later than 2 hours after the beginning of contacting the population of cells with the one or more cytokines described above.

[0345] In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR(s) occurs no later than 1 hour after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, the population of cells is harvested for storage or administration. In some embodiments, the population of cells is harvested for storage or administration no later than 72, 60, 48, 36, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, or 18 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, the population of cells is harvested for storage or administration no later than 26 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, the population of cells is harvested for storage or administration no later than 25 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, the population of cells is harvested for storage or administration no later than 24 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, the population of cells is harvested for storage or administration no later than 106ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 23 hours after the beginning of contacting the population of cells with the one or more cytokines described above. In some embodiments, the population of cells is harvested for storage or administration no later than 22 hours after the beginning of contacting the population of cells with the one or more cytokines described above.

[0346] In some embodiments, the population of cells is not expanded ex vivo. In some embodiments, the population of cells is expanded by no more than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, or 60%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 5%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 10%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 15%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 20%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 25%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 30%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 35%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. In some embodiments, the population of cells is expanded by no more than 40%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above.

[0347] In some embodiments, the population of cells is expanded by no more than 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 11, 12, 16, 20, 24, 36, or 48 hours, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the one or more cytokines described above. 107ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0348] In some embodiments, the population of cells is not contacted in vitro with an agent that stimulates a CD3 / TCR complex (for example, an anti-CD3 antibody) and / or an agent that stimulates a costimulatory molecule on the surface of the cells (for example, an anti-CD28 antibody), or if contacted, the contacting step is less than 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 hours.

[0349] In some embodiments, the population of cells is contacted in vitro with an agent that stimulates a CD3 / TCR complex (for example, an anti-CD3 antibody) and / or an agent that stimulates a costimulatory molecule on the surface of the cells (for example, an anti-CD28 antibody) for 20, 21, 22, 23, 24, 25, 26, 27, or 28 hours.

[0350] In some embodiments, the population of cells manufactured using the cytokine process provided herein shows a higher percentage of naive cells among CAR-expressing cells (for example, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, or 60% higher), compared with cells made by an otherwise similar method which further comprises contacting the population of cells with, for example, an agent that binds a CD3 / TCR complex (for example, an anti-CD3 antibody) and / or an agent that binds a costimulatory molecule on the surface of the cells (for example, an anti-CD28 antibody).

[0351] In some embodiments, the cytokine process provided herein is conducted in cell media comprising no more than 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, or 8% serum. In some embodiments, the cytokine process provided herein is conducted in cell media comprising a LSD1 inhibitor, a MALT1 inhibitor, or a combination thereof.

[0352] Elutriation

[0353] In some embodiments, the methods described herein feature an elutriation method that removes unwanted cells, for example, monocytes and blasts, thereby resulting in an improved enrichment of desired immune effector cells suitable for CAR expression. In some embodiments, the elutriation method described herein is optimized for the enrichment of desired immune effector cells suitable for CAR expression from a previously frozen sample, for example, a thawed sample. In some embodiments, the elutriation method described herein provides a preparation of cells with improved purity as compared to a preparation of cells collected from the elutriation protocols known in the art. In some embodiments, the elutriation method described herein includes using an optimized viscosity of the starting sample, for example, cell sample, for example, thawed cell sample, by dilution with certain isotonic solutions (for example, PBS), and using an optimized combination of flow rates and collection volume for each fraction collected by an elutriation device. Exemplary elutriation methods that 108ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION could be applied in the present disclosure are described on pages 48-51 of WO 2017 / 117112, herein incorporated by reference in its entirety. Density Gradient Centrifugation

[0354] Manufacturing of adoptive cell therapeutic product requires processing the desired cells, for example, immune effector cells, away from a complex mixture of blood cells and blood elements present in peripheral blood apheresis starting materials. Peripheral blood- derived lymphocyte samples have been successfully isolated using density gradient centrifugation through Ficoll solution. However, Ficoll is not a preferred reagent for isolating cells for therapeutic use, as Ficoll is not qualified for clinical use. In addition, Ficoll contains glycol, which has toxic potential to the cells. Furthermore, Ficoll density gradient centrifugation of thawed apheresis products after cryopreservation yields a suboptimal T cell product, for example, as described in the Examples herein. For example, a loss of T cells in the final product, with a relative gain of non-T cells, especially undesirable B cells, blast cells and monocytes was observed in cell preparations isolated by density gradient centrifugation through Ficoll solution.

[0355] Without wishing to be bound by theory, it is believed that immune effector cells, for example, T cells, dehydrate during cryopreservation to become denser than fresh cells. Without wishing to be bound by theory, it is also believed that immune effector cells, for example, T cells, remain denser longer than the other blood cells, and thus are more readily lost during Ficoll density gradient separation as compared to other cells. Accordingly, without wishing to be bound by theory, a medium with a density greater than Ficoll is believed to provide improved isolation of desired immune effector cells in comparison to Ficoll or other mediums with the same density as Ficoll, for example, 1.077 g / mL.

[0356] In some embodiments, the density gradient centrifugation method described herein includes the use of a density gradient medium comprising iodixanol. In some embodiments, the density gradient medium comprises about 60% iodixanol in water.

[0357] In some embodiments, the density gradient centrifugation method described herein includes the use of a density gradient medium having a density greater than Ficoll. In some embodiments, the density gradient centrifugation method described herein includes the use of a density gradient medium having a density greater than 1.077 g / mL, for example, greater than 1.077 g / mL, greater than 1.1 g / mL, greater than 1.15 g / mL, greater than 1.2 g / mL, greater than 1.25 g / mL, greater than 1.3 g / mL, greater than 1.31 g / mL. In some embodiments, the density gradient medium has a density of about 1.32 g / mL. Additional embodiments of density gradient 109ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION centrifugation are described on pages 51-53 of WO 2017 / 117112, herein incorporated by reference in its entirety.

[0358] Enrichment by Selection

[0359] Provided herein are methods for selection of specific cells to improve the enrichment of the desired immune effector cells suitable for CAR expression. In some embodiments, the selection comprises a positive selection, for example, selection for the desired immune effector cells. In some embodiments, the selection comprises a negative selection, for example, selection for unwanted cells, for example, removal of unwanted cells. In some embodiments, the positive or negative selection methods described herein are performed under flow conditions, for example, by using a flow-through device, for example, a flow-through device described herein. Exemplary positive and negative selections are described on pages 53-57 of WO 2017 / 117112, herein incorporated by reference in its entirety. Selection methods can be performed under flow conditions, for example, by using a flow- through device, also referred to as a cell processing system, to further enrich a preparation of cells for desired immune effector cells, for example, T cells, suitable for CAR expression. Exemplary flow-through devices are described on pages 57- 70 of WO 2017 / 117112, herein incorporated by reference in its entirety. Exemplary cell separation and debeading methods are described on pages 70-78 of WO 20l 7 / 117112, herein incorporated by reference in its entirety.

[0360] Selection procedures are not limited to ones described on pages 57-70 of WO 2017 / 117112. Negative T cell selection via removal of unwanted cells with CD19, CD14 and CD26 Miltenyi beads in combination with column technology (CliniMACS® Plus or CliniMACS® Prodigy®) or positive T cell selection with a combination of CD4 and CD8 Miltenyi beads and column technology (CliniMACS® Plus or CliniMACS® Prodigy®) can be used. Alternatively, column-free technology with releasable CD3 beads (GE Healthcare) can be used. In addition, bead-free technologies such as ThermoGenesis X-series devices can be utilized as well.

[0361] Methods of making a cell disclosed herein include those known in the art, e.g., as described in CN 108103105, CN 108085342, CN 108018312, CN 107287164, WO 18052947, WO 17123956, WO 17114497, WO 17103596, WO 17068421, WO 17023803, WO 17015427, WO 16196388, WO 16168595, WO 14186469, WO 17165245, WO 18106732, WO 17015490, WO 18075813, WO 18102761, WO 17127755, WO 17214333, WO 18059549, WO 17190100, WO 16180778, WO 18057823, and / or CN 106957822, each one of which is hereby incorporated by reference in its entirety.

[0362] Sources of Cells 110ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0363] Prior to expansion and genetic modification or other modification, a source of cells, e.g., T cells or natural killer (NK) cells, can be obtained from a subject. The term “subject” is intended to include living organisms in which an immune response can be elicited (e.g., mammals). Examples of subjects include humans, monkeys, chimpanzees, dogs, cats, mice, rats, and transgenic species thereof. T cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In certain embodiments of the present disclosure disclosure, immune effector cells, e.g., T cells, can be obtained from a unit of blood collected from a subject using any number of techniques known to the skilled artisan, such as Ficoll™ separation. In one preferred embodiment, cells from the circulating blood of an individual are obtained by apheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets. In some embodiments, the cells collected by apheresis may be washed to remove the plasma fraction and, optionally, to place the cells in an appropriate buffer or media for subsequent processing steps. In one embodiment of the invention, the cells are washed with phosphate buffered saline (PBS). In an alternative embodiment, the wash solution lacks calcium and may lack magnesium or may lack many if not all divalent cations. Initial activation steps in the absence of calcium can lead to magnified activation. As those of ordinary skill in the art would readily appreciate a washing step may be accomplished by methods known to those in the art, such as by using a semi- automated “flow-through” centrifuge (for example, the Cobe 2991 cell processor, the Baxter CytoMate, or the Haemonetics Cell Saver 5) according to the manufacturer’s instructions. After washing, the cells may be resuspended in a variety of biocompatible buffers, such as, for example, Ca-free, Mg-free PBS, PlasmaLyte A, or other saline solution with or without buffer. Alternatively, the undesirable components of the apheresis sample may be removed and the cells directly resuspended in culture media.

[0364] In some embodiments, T cells are isolated from peripheral blood lymphocytes by lysing the red blood cells and depleting the monocytes, for example, by centrifugation through a PERCOLLTMgradient or by counterflow centrifugal elutriation. The methods described herein can include, e.g., selection of a specific subpopulation of immune effector cells, e.g., T cells, that are a T regulatory cell-depleted population, CD25+ depleted cells, using, e.g., a negative selection technique, e.g., described herein. Preferably, the population of T regulatory depleted cells contains less than 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1% of CD25+ cells. 111ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0365] In some embodiments, T regulatory cells, e.g., CD25+ T cells, are removed from the population using an anti-C25 antibody, or fragment thereof, or a CD25-binding ligand, IL- 2. In some embodiments, the anti-CD25 antibody, or fragment thereof, or CD25-binding ligand is conjugated to a substrate, e.g., a bead, or is otherwise coated on a substrate, e.g., a bead. In some embodiments, the anti-CD25 antibody, or fragment thereof, is conjugated to a substrate as described herein.

[0366] In some embodiments, the T regulatory cells, e.g., CD25+ T cells, are removed from the population using CD25 depletion reagent from Militenyi™. In some embodiments, the ratio of cells to CD25 depletion reagent is le7 cells to 20 uL, or le7 cells to 15 uL, or le7 cells to 10 uL, or le7 cells to 5 uL, or le7 cells to 2.5 uL, or le7 cells to 1.25 uL.

[0367] In some embodiments, the population of immune effector cells to be depletedincludes about 6 x 109CD25+ T cells. In other embodiments, the population of immune effectorcells to be depleted include about 1 x 109to 1x 1010CD25+ T cell, and any integer value inbetween. In some embodiments, the resulting population T regulatory depleted cells has 2 x109T regulatory cells, e.g., CD25+ cells, or less (e.g., 1 x 109, 5 x 108, 1 x 108, 5 x 107, 1 x 107,or less CD25+ cells).

[0368] In some embodiments, the T regulatory cells, e.g., CD25+ cells, are removed from the population using the CliniMAC system with a depletion tubing set, such as, e.g., tubing 162-01. In some embodiments, the CliniMAC system is run on a depletion setting such as, e.g., DEPLETION2. l.

[0369] The methods described herein can include more than one selection step, e.g., more than one depletion step. Enrichment of a T cell population by negative selection can be accomplished, e.g., with a combination of antibodies directed to surface markers unique to the negatively selected cells. One method is cell sorting and / or selection via negative magnetic immunoadherence or flow cytometry that uses a cocktail of monoclonal antibodies directed to cell surface markers present on the cells negatively selected. For example, to enrich for CD4+ cells by negative selection, a monoclonal antibody cocktail can include antibodies to CD14, CD20, CD11b, CD16, HLA-DR, and CD8.

[0370] Also provided are methods that include removing cells from the population which express a check point inhibitor, e.g., a check point inhibitor described herein, e.g., one or more of PD1 + cells, LAG3+ cells, and TIM3+ cells, to thereby provide a population of T regulatory depleted, e.g., CD25+ depleted cells, and check point inhibitor depleted cells, e.g., PD1 +, LAG3+ and / or TIM3+ depleted cells. Exemplary check point inhibitors include B7-Hl, B&-1, 112ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION CD160, PIH, 2B4, PDI, TIM3, CEACAM (e.g., CEACAM-1, CEACAM-3 and / or CEACAM- 5), LAG3, TIGIT, CTLA-4, BTLA and LAIRI. In some embodiments, check point inhibitor expressing cells are removed simultaneously with the T regulatory, e.g., CD25+ cells. For example, an anti-C25 antibody, or fragment thereof, and an anti-check point inhibitor antibody, or fragment thereof, can be attached to the same bead which can be used to remove the cells, or an anti-CD25 antibody, or fragment thereof, and the anti-check point inhibitor antibody, or fragment there, can be attached to separate beads, a mixture of which can be used to remove the cells. In some embodiments, the removal of T regulatory cells, e.g., CD25+ cells, and the removal of the check point inhibitor expressing cells is sequential, and can occur, e.g., in either order.

[0371] Methods described herein can include a positive selection step. For example, T cells can be isolated by incubation with anti-CD3 / anti-CD28 (e.g., 3x28)-conjugated beads, such as DYNABEADS® M-450 CD3 / CD28 T, for a time period sufficient for positive selection of the desired T cells. In some embodiments, the time period is about 30 minutes. In a further embodiment, the time period ranges from 30 minutes to 36 hours or longer and all integer values there between. In a further embodiment, the time period is at least 1, 2, 3, 4, 5, or 6 hours. In yet another preferred embodiment, the time period is 10 to 24 hours. In some embodiments, the incubation time period is 24 hours. Longer incubation times may be used to isolate T cells in any situation where there are few T cells as compared to other cell types, such in isolating tumor infiltrating lymphocytes (TIL) from tumor tissue or from immunocompromised individuals. Further, use of longer incubation times can increase the efficiency of capture of CD8+ T cells. Thus, by simply shortening or lengthening the time T cells are allowed to bind to the CD3 / CD28 beads and / or by increasing or decreasing the ratio of beads to T cells (as described further herein), subpopulations of T cells can be preferentially selected for or against at culture initiation or at other time points during the process. Additionally, by increasing or decreasing the ratio of anti-CD3 and / or anti-CD28 antibodies on the beads or other surface, subpopulations of T cells can be preferentially selected for or against at culture initiation or at other desired time points.

[0372] In some embodiments, a T cell population can be selected that expresses one or more of IFN-r, TNFa, IL-l 7A, IL-2, IL-3, IL-4, GM-CSF, IL-10, IL-13, granzyme B, and perforin, or other appropriate molecules, e.g., other cytokines. Methods for screening for cell expression can be determined, e.g., by the methods described in PCT Publication No.: WO 2013 / 126712. 113ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION

[0373] For isolation of a desired population of cells by positive or negative selection, the concentration of cells and surface (e.g., particles such as beads) can be varied. In certain embodiments, it may be desirable to significantly decrease the volume in which beads and cells are mixed together (e.g., increase the concentration of cells), to ensure maximum contact of cells and beads. For example, In some embodiments, a concentration of about 10 billion cells / ml, 9 billion / ml, 8 billion / ml, 7 billion / ml, 6 billion / ml, or 5 billion / ml is used. In some embodiments, a concentration of 1 billion cells / ml is used. In some embodiments, a concentration of cells from 75, 80, 85, 90, 95, or 100 million cells / ml is used. In further embodiments, concentrations of 125 or 150 million cells / ml can be used.

[0374] Using high concentrations can result in increased cell yield, cell activation, and cell expansion. Further, use of high cell concentrations allows more efficient capture of cells that may weakly express target antigens of interest, such as CD28-negative T cells, or from samples where there are many tumor cells present (e.g., leukemic blood, tumor tissue, etc.). Such populations of cells may have therapeutic value and would be desirable to obtain. For example, using high concentration of cells allows more efficient selection of CD8+ T cells that normally have weaker CD28 expression.

[0375] In a related embodiment, it may be desirable to use lower concentrations of cells. By significantly diluting the mixture of T cells and surface (e.g., particles such as beads), interactions between the particles and cells is minimized. This selects for cells that express high amounts of desired antigens to be bound to the particles. For example, CD4+ T cells express higher levels of CD28 and are more efficiently captured than CD8+ T cells in diluteconcentrations. In some embodiments, the concentration of cells used is 5 X 106 / ml. In otherembodiments, the concentration used can be from about 1 X 105 / ml to 1 X 106 / ml, and any integer value in between.

[0376] In other embodiments, the cells may be incubated on a rotator for varying lengths of time at varying speeds at eith...

Claims

ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION What is claimed is:

1. A method of treating a viral disorder, comprising administering to a patient in need thereof a pharmaceutical composition comprising a therapeutically effective amount of cells comprising a nucleic acid molecule encoding a chimeric antigen receptor (CAR) molecule that binds to cells expressing at least one of an EBV antigen, an SL9 antigen, a Nef antigen, or a SARS antigen.

2. The method of claim 1, wherein the CAR molecule binds to a cell expressing at least one of (1) the EBV antigen and an HLA molecule comprising HLA-A*02, or a functional variant thereof, (2) the SL9 antigen and an HLA molecule comprising HLA-A*02, or a functional variant thereof, (3) the Nef antigen and an HLA molecule comprising HLA-A*24, or a functional variant thereof, or (4) the SARS antigen and an HLA molecule comprising HLA-B*08, or a functional variant thereof.

3. The method of claim 1 or 2, wherein the nucleic acid molecule comprises a first nucleic acid sequence encoding an EBV-A02 binding peptide comprising one selected from the group consisting of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), and SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof 159ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50).

4. The method of claim 1 or 2, wherein the nucleic acid molecule comprises a first nucleic acid sequence encoding a SL9-A02 binding peptide comprising one selected from the group consisting of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), 160ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), and YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100).

5. The method of claim 1 or 2, wherein the nucleic acid molecule comprises a first nucleic acid sequence encoding a Nef_A24 binding peptide comprising one selected from the group consisting of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID 161ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), and WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150).

6. The method of claim 1 or 2, wherein the nucleic acid molecule comprises a first nucleic acid sequence encoding a SARS-B08 binding peptide comprising one selected from the group consisting of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 162ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), and VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200).

7. The method of any of claims 1 through 6, wherein the first nucleic acid sequence encodes a peptide comprising GTHDKCENPKX1X2X3X4X5X6X7X8QNX9NVVX10TNKELEDIYDESN (SEQ ID NO: 202), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 202, wherein X1X2X3X4X5X6 X7X8 is one of SEQ ID NOS: 1 through 200 or a functional variant thereof comprising about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200, and X9 and X10 are any amino acid. 163ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 8. The method of any one of claims 1 through 7, wherein the nucleic acid molecule further comprises a second nucleic acid sequence encoding a peptide comprising KEETKEVLKKFKEKVNQFX11EHAFDIINKYGDKEIX12NMMX13MLLWRVX14FRSFRI DANNVELIKFNIRVLDWIMAEADNDLSYFISQ (SEQ ID NO: 203), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 203, wherein X11, X12, X13, and X14 are any amino acid.

9. The method of any one of claims 1 through 8, wherein the nucleic acid molecule comprises the first nucleic acid sequence and a third nucleic acid sequence encoding a single chain antibody or antibody fragment that binds to a cell expressing CD3, the first nucleic acid sequence and the third nucleic acid sequence together encode a chimeric peptide comprising a first domain and a second domain, the first domain comprising the peptide encoded by the first nucleic acid sequence, and the second domain comprising the single chain antibody or antibody fragment that binds to a cell expressing CD3.

10. The method of claim 9, wherein the nucleic acid molecule further comprises the second nucleic acid sequence, and the first domain comprises the peptide encoded by the first nucleic acid sequence and the peptide encoded by the second nucleic acid sequence.

11. The method of any one of claims 1 through 10, wherein the therapeutically effective amount is about 3x104cells / kg body weight for patients with less than or equal to 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition.

12. The method of any of claims 1 through 10, wherein the therapeutically effective amount is from about 1.5 x 106cells for patients with more than 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition.

13. The method of any of claims 1 through 12, wherein the viral disorder comprises a cell expressing one or more of an EBV antigen, an SL9 antigen, a Nef antigen, and a SARS antigen.

14. The method of claim 12, wherein the viral disorder comprises a cell expressing one or more of (1) an EBV antigen and an HLA molecule comprising HLA-A*02, or a functional variant thereof, (2) an SL9 antigen and an HLA molecule comprising HLA-A*02, or a functional variant 164ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION thereof, (3) a Nef antigen and an HLA molecule comprising HLA-A*24, or a functional variant thereof, or (4) a SARS antigen and an HLA molecule comprising HLA-B*08, or a functional variant thereof.

15. The method any one of claims 1 through 14, wherein the viral disorder is one differentiated by having an EBV antigen as a biomarker therefor.

16. The method of claim 14 or 15, wherein the viral disorder is infection with an Epstein- Barr virus, a viral disorder caused by an Epstein-Barr virus, or a cancer caused by an Epstein- Barr virus.

17. The method of any one of claim 1 through 14, wherein the viral disorder is one differentiated by having an SL9 antigen as a biomarker therefor.

18. The method any one of claims 1 through 14 or 17, wherein the viral disorder is infection with human immunodeficiency virus 1 (HIV-1) or acquired immune deficience syndrome.

19. The method any one of claims 1 through 14, wherein the viral disorder is one differentiated by having a Nef antigen as a biomarker therefor.

20. The method any one of claims 1 through 14 or 19, wherein the viral disorder is infection with human immunodeficiency virus 1 (HIV-1) or acquired immune deficience syndrome.

21. The method any one of claims 1 through 14, wherein the viral disorder is one differentiated by having a SARS antigen as a biomarker therefor.

22. The method any one of claims 1 through 14 or 21, wherein the viral disorder is infection by SARS-Cov2 or COVID-19.

23. The method of any of claims 1 through 22, wherein the therapeutically effective amount is from about 3.0 x 105to 1.5 x l06cells.

24. The method of any of claims 1 through 22, wherein the therapeutically effective amount is about 10 x l04cells / kg body weight for patients with less than or equal to about 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition. 165ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 25. The method of any of claims 1 through 22, wherein the therapeutically effective amount is from about 5 x106cells for patients with more than about 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition.

26. The method of any of claims 1 through 22, wherein the therapeutically effective amount is about 3.0x l04cells / kg body weight for patients with less than or equal to 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition.

27. The method of any one of claims 1 through 22, wherein the therapeutically effective amount is about 1.5 x 106cells for patients with more than 50 kg body weight based on the total CAR-T cells in the pharmaceutical composition.

28. The method of any one of claims 1 through 22, wherein the therapeutically effective amount is about 1x104cells / kg body weight, 2x 104cells / kg body weight, 3 x 104cells / kg body weight, 4 x 104cells / kg body weight, 5 x104cells / kg body weight, 6 x104cells / kg body weight, 7 x104cells / kg body weight, 8 x104cells / kg body weight, 9 x104cells / kg body weight, 10 x104cells / kg body weight, 11 x 104cells / kg body weight, 12 x 104cells / kg body weight, 13 x 104cells / kg body weight, 14 x 104cells / kg body weight, 15 x 104cells / kg body weight, 16 x 104cells / kg body weight, 17 x 104cells / kg body weight, 18 x 104cells / kg body weight, 19 x 104cells / kg body weight, 20 x 104cells / kg body weight, 21 x 104cells / kg body weight, 22 x 104cells / kg body weight, 23 x 104cells / kg body weight, 24 x 104cells / kg body weight, 25 x 104cells / kg body weight, 26 x 104cells / kg body weight, 27 x 104cells / kg body weight, 28 x 104cells / kg body weight, 29 x 104cells / kg body weight, 30 x 104cells / kg body weight, 40 x 104cells / kg body weight, 50 x 104cells / kg body weight, 60 x 104cells / kg body weight, 70 x 104cells / kg body weight, 80 x 104cells / kg body weight, or about 90 x 104cells / kg body weight.

29. The method of any one of claims 1 through 22, wherein the therapeutically effective amount is about 0.5 x 106cells, 1 x106cells, 1. 5 x 106cells, 2 x 106cells, 2.5 x106cells, 3 x106cells, 3.5 x 106cells, 4 x106cells, 4.5 x 106cells, 5 x 106cells, 5.5 x 106cells, 6 x 106cells, 6.5 x 106cells, 7 x 106cells, 7.5 x 106cells, 8 x 106cells, 8.5 x 106cells, 9 x 106cells, 9.5 x 106cells, 10 x 106cells, 10.5 x 106cells, 11 x 106cells, 11.5 x 106cells, 12 x 106cells, 12.5 x106cells, 13 x 106cells, 13.5 x106cells, 14 x106cells,14.5 xl 06cells, 15 x 106cells, 16 x 106cells, 17 x 106cells, 18 x 106cells, 19 x 106cells, 20 x 106cells, 2l x 106cells, 22 x 166ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 106cells, 23 x 106cells, 24 x 106cells, 25 x 106cells, 26 xl 06cells, 27 x 106cells, 28x106cells, 29 x 106cells, 30 x 106cells, 31 x 106cells, 32 x 106cells, 33 x 106cells, 34x106cells, 35 x 106cells, 36 x 106cells, 37 x 106cells, 38 x 106cells, 39 x 106cells, 40 x 106cells, 41 x 106cells,42x 106cells, 43 x 106cells, 44 x 106cells, or 45 x 106cells.

30. The method of any one of claims 1 through 29, wherein the first nucleic acid sequence encodes a peptide comprising an amino acid sequence with at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence of one of SEQ ID NOS: 1 through 200.

31. A cell comprising a CAR molecule comprising an amino acid sequence that binds to a cell expressing at least one of (1) an EBV antigen and an HLA molecule comprising HLA- A*02, or a functional variant thereof, (2) an SL9 antigen and an HLA molecule comprising HLA-A*02, or a functional variant thereof, (3) a Nef antigen and an HLA molecule comprising HLA-A*24, or a functional variant thereof, or (4) a SARS antigen and an HLA molecule comprising HLA-B*08, or a functional variant thereof.

32. A cell comprising a nucleic acid molecule comprising a first nucleic acid sequence encoding an amino acid sequence comprising one or a combination of two or more of: EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), 167ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50).

33. A cell comprising a nucleic acid molecule comprising a first nucleic acid sequence encoding one or a combination of two or more of: YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG 168ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100).

34. A cell comprising a nucleic acid molecule comprising a first nucleic acid sequence encoding one or a combination of two or more of: WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), 169ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150).

35. A cell comprising a nucleic acid molecule comprising a nucleic acid sequence encoding one or a combination of two or more of: KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), 170ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200).

36. The cell of claim and one of claims 32 through 35, wherein the first nucleic acid sequence encodes a peptide comprising GTHDKCENPKX1X2X3X4X5X6X7X8QNX9NVVX10TNKELEDIYDESN (SEQ ID NO:202), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 202, wherein X1X2X3X4X5X6X7X8 is one of SEQ ID NOS: 1 through 200 or a functional variant thereof comprising about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200, and X9and X10are any amino acid. 171ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 37. The cell of any one of claims 32 through 36, wherein the nucleic acid molecule further comprises a second nucleic acid sequence encoding a peptide comprising KEETKEVLKKFKEKVNQFX11EHAFDIINKYGDKEIX12NMMX13MLLWRVX14FRSFRI DANNVELIKFNIRVLDWIMAEADNDLSYFISQ (SEQ ID NO:203), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 203, wherein X11, X12, X13, and X14are any amino acid.

38. The cell of any of claims 32 through 37, wherein the nucleic acid molecule comprises the first nucleic acid sequence and a third nucleic acid sequence encoding a single chain antibody or antibody fragment that binds to a cell expressing CD3, the first nucleic acid sequence and the third nucleic acid sequence together encode a chimeric peptide comprising a first domain and a second domain, the first domain comprising the peptide encoded by the first nucleic acid sequence, and the second domain comprising the single chain antibody or antibody fragment that binds to a cell expressing CD3.

39. The cell of claim 38, wherein the nucleic acid molecule further comprises the second nucleic acid sequence, and the first domain comprises the peptide encoded by the first nucleic acid sequence and the peptide encoded by the second nucleic acid sequence.

40. The cell of any of claims 32 through 39, wherein the cells is a T cell or an NK cell.

41. The cell of any of claims 32 through 40, wherein the cell further comprises one or more further nucleic acid sequences encoding one or more of a CD8α hinge-transmembrane domain, a 41BB co-stimulatory domain and a CD3ζ signaling domain.

42. The cell of claim 41 wherein the nucleic acid molecule comprises the first nucleic sequence, the second nucleic acid sequence, and the third nucleic acid sequence, respectively encoding (1) one of SEQ ID NOS:1 through 200 or SEQ ID NO: 202 and (2) SEQ ID NO: 203, or a functional variant thereof comprising from about 75% sequence identity to (1) one of SEQ ID NOS:1 through 200 or SEQ ID NO: 202 and (2) SEQ ID NO: 203, respectively.

43. A pharmaceutical composition comprising a therapeutically effective amount of one or a combination of two or more of: EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), 172ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50); and a pharmaceutically acceptable carrier. 173ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 44. A pharmaceutical composition comprising a therapeutically effective amount of one or a combination of two or more of: YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), 174ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100); and a pharmaceutically acceptable carrier.

45. A pharmaceutical composition comprising a therapeutically effective amount of one or a combination of two or more of: WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID 175ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150); and a pharmaceutically acceptable carrier.

46. A pharmaceutical composition comprising a therapeutically effective amount of one or a combination to two or more of: KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN 176ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200); and a pharmaceutically acceptable carrier.

47. The pharmaceutical composition of any one of claims 42 through 46 further comprising a cell that comprises one of SEQ ID NOS:1 through 200, or a functional variant thereof comprising from about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200.

48. The pharmaceutical composition of any one of claims 42 through 47, wherein the peptide comprises one or a combination of (1) GTHDKCENPKX1X2X3X4X5X6 X7X8QNX9NVVX10TNKELEDIYDESN (SEQ ID NO:202), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO:202, wherein X1X2X3X4X5X6 X7X8is one of SEQ ID NOS: 1 through 200 or a functional variant thereof comprising about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200, and X9 and X10 are any amino acid, and (2) KEETKEVLKKFKEKVNQFX11EHAFDIINKYGDKEIX12NMMX13MLLWRVX14FRSFRI DANNVELIKFNIRVLDWIMAEADNDLSYFISQ (SEQ ID NO:203), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 203, wherein X11, X12, X13, and X14are any amino acid.

49. The pharmaceutical composition of any of claims 42 through 48, wherein the peptide further comprises one or a combination of: a CD8α hinge-transmembrane domain, a 41BB co- stimulatory domain and a CD3ζ signaling domain.

50. A pharmaceutical composition comprising a therapeutically effective amount of a cell comprising one or more peptide selected from: EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), 177ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), and SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50); and a pharmaceutically acceptable carrier.

51. A pharmaceutical composition comprising a therapeutically effective amount of a cell comprising one or more peptide selected from: YMNLYNFG (SEQ ID NO: 51), QYELSNQL 178ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100); and a pharmaceutically acceptable carrier. 179ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 52. A pharmaceutical composition comprising a therapeutically effective amount of a cell comprising one or more peptide selected from WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), and WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), 180ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150); and a pharmaceutically acceptable carrier.

53. A pharmaceutical composition comprising a therapeutically effective amount of a cell comprising one or more peptide selected from KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), and VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), 181ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200); and a pharmaceutically acceptable carrier.

55. The pharmaceutical composition of any one of claims 43 through 54, wherein the peptide comprises an amino acid sequence comprising one or a combination of SEQ ID NO:202 and SEQ ID NO:203, or a functional variant thereof comprising an amino acid sequence comprising from about 75% sequence identity to about 99% sequence identity to SEQ ID NO:202 and SEQ ID NO:203, respectively.

56. The pharmaceutical composition of any one of claims 43 or 55, wherein the peptide further comprises a CD8α hinge-transmembrane domain, a 41BB co-stimulatory domain and a CD3ζ signaling domain.

56. A pharmaceutical composition comprising the cell of any one of claims 31 through 42 and a pharmaceutically acceptable carrier.

57. A kit comprising the cell of any one of claims 31 through 42 or the pharmaceutical composition of any one of claims 43 through 56.

58. A nucleic acid molecule comprising a first nucleic acid sequence encoding an amino acid sequence comprising one or a combination of two or more of: EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), 182ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50).

59. A nucleic acid molecule comprising a first nucleic acid sequence encoding one or a combination of two or more of: YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), 183ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100).

60. A nucleic acid molecule comprising a first nucleic acid sequence encoding one or a combination of two or more of: WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM 184ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150).

61. A nucleic acid molecule comprising a first nucleic acid sequence encoding one or a combination of two or more of: KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), 185ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200).

62. A nucleic acid molecule comprising a nucleic acid sequence encoding a peptide comprising one or a combination of (1) GTHDKCENPKX1X2X3X4X5X6 X7X8QNX9NVVX10TNKELEDIYDESN (SEQ ID NO:202), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO:202, wherein X1X2X3X4X5X6 186ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION X7X8 is one of SEQ ID NOS: 1 through 200 or a functional variant thereof comprising about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200, and X9and X10are any amino acid, and (2) KEETKEVLKKFKEKVNQFX11EHAFDIINKYGDKEIX12NMMX13MLLWRVX14FRSFRI DANNVELIKFNIRVLDWIMAEADNDLSYFISQ (SEQ ID NO:203), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 203, wherein X11, X12, X13, and X14 are any amino acid.

63. A kit comprising (1) a nucleic acid herein or the nucleic acid of any one of claims 58 through 62 and (2) a cell capable of being transformed by the nucleic acid.

64. A peptide comprising an amino acid sequence comprising one or a combination of two or more of: EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50), or a functional variant thereof comprising about 85% sequence identity to a respective one of EWFETYVH (SEQ ID NO: 1), DLFWIGAD(SEQ ID NO: 2), DLFETYVH(SEQ ID NO: 3), EWFWIGAD(SEQ ID NO: 4), TIWWIGRD(SEQ ID NO: 5), TIWETYVH(SEQ ID NO: 6), EWFWTYVH(SEQ ID NO: 7), EWFETYFH(SEQ ID NO: 8), EWFWIGRD(SEQ ID NO: 9), EWFETYVN(SEQ ID NO: 10), EWFQTYVH(SEQ ID NO: 11), PWNETYVH(SEQ ID NO: 12), EYFETYVH(SEQ ID NO: 13), PWNWIGAY(SEQ ID NO: 14), EYFWIGKD(SEQ ID NO: 15), EWFEIGAD(SEQ ID NO: 16), 187ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION EWFETDVH(SEQ ID NO: 17), EQIETYVH(SEQ ID NO: 18), EQIWIAHD(SEQ ID NO: 19), VFSETYVH(SEQ ID NO: 20), TENYIAND(SEQ ID NO: 21), TENETYVH(SEQ ID NO: 22), DWFETYVH(SEQ ID NO: 23), EWFWIGAY(SEQ ID NO: 24), EWFET_VH(SEQ ID NO: 25), DLFWTYVH(SEQ ID NO: 26), TWFETYVH(SEQ ID NO: 27), EWFWIGKD(SEQ ID NO: 28), HWGETYVH(SEQ ID NO: 29), VFSYIGYQ(SEQ ID NO: 30), EWFETYDH(SEQ ID NO: 31), YTLWIGHD(SEQ ID NO: 32), KHGETYVH(SEQ ID NO: 33), EWFETYVA(SEQ ID NO: 34), EWFWIAHD(SEQ ID NO: 35), EWFYTYVH(SEQ ID NO: 36), YTLETYVH(SEQ ID NO: 37), AGYSTYLY(SEQ ID NO: 38), EWFEIGRD(SEQ ID NO: 39), MTEWIARD(SEQ ID NO: 40), ESFWIGGD(SEQ ID NO: 41), KDGDTVYY(SEQ ID NO: 42), MPDYIGWD(SEQ ID NO: 43), DAPYIGDT(SEQ ID NO: 44), NFFWLGQD(SEQ ID NO: 45), EYDVIGRG(SEQ ID NO: 46), VANWIGRD(SEQ ID NO: 47), SIYWTGQD(SEQ ID NO: 48), QVTYIGYG(SEQ ID NO: 49), or SYFWLAWD (SEQ ID NO: 50).

65. A peptide comprising an amino acid sequence comprising one or a combination of two or more of: YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100), or a functional variant thereof comprising about 85% sequence identity to a respective one of YMNLYNFG (SEQ ID NO: 51), QYELSNQL (SEQ ID NO: 52), QYELYNFG (SEQ ID NO: 53), YMNLSNQL (SEQ ID NO: 54), YMNLYNQL (SEQ ID NO: 55), YMNLYNFC (SEQ ID NO: 56), QYELSNFG (SEQ ID NO: 57), YMNLYNFV (SEQ ID NO: 58), YMNLSNFG (SEQ ID NO: 59), YMNLYTFG (SEQ ID NO: 60), YMNLYNVG (SEQ ID NO: 61), YMTLMYFG (SEQ ID NO: 62), YMTLYNFG (SEQ ID NO: 63), 188ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION YMNLMYFG (SEQ ID NO: 64), YMNLYNLG (SEQ ID NO: 65), QYNLYNFG (SEQ ID NO: 66), YMELSNQL (SEQ ID NO: 67), QMNLYNFG (SEQ ID NO: 68), YMNLYIFG (SEQ ID NO: 69), YMELYNFG (SEQ ID NO: 70), YMNLYNYG (SEQ ID NO: 71), YMNLYHFG (SEQ ID NO: 72), YMNLYKFG (SEQ ID NO: 73), YYELYNFG (SEQ ID NO: 74), YYELSNQL (SEQ ID NO: 75), YMNLYYFG (SEQ ID NO: 76), YMNLFNFG (SEQ ID NO: 77), NMNLYNFG (SEQ ID NO: 78), YMNWTTSV (SEQ ID NO: 79), YMNLDNFG (SEQ ID NO: 80), QYELMYFG (SEQ ID NO: 81), YINLYNFG (SEQ ID NO: 82), YMKLYNFG (SEQ ID NO: 83), YMNLYNSG (SEQ ID NO: 84), HMNLYNFG (SEQ ID NO: 85), YMNLYNFF (SEQ ID NO: 86), YMDLYNFG (SEQ ID NO: 87), QYELYNQL (SEQ ID NO: 88), YMNLHNFG (SEQ ID NO: 89), YMSLYNFG (SEQ ID NO: 90), YKNLYNFG (SEQ ID NO: 91), YMNLNNFG (SEQ ID NO: 92), QYELSNQM (SEQ ID NO: 93), YMNLYNIG (SEQ ID NO: 94), QYELYNFC (SEQ ID NO: 95), YMNLYNCG (SEQ ID NO: 96), YMNLYNFA (SEQ ID NO: 97), QYELSTQL (SEQ ID NO: 98), QYELYNFV (SEQ ID NO: 99), or YMNLYNFD (SEQ ID NO: 100).

66. A peptide comprising an amino acid sequence comprising one or a combination of two or more of: WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150), or a functional variant thereof comprising about 85% sequence identity to a respective one of WTNNFGKT (SEQ ID NO: 101), WTNNWGVT (SEQ ID NO: 102), WYNTYGKM (SEQ ID NO: 103), WTNTYGKM (SEQ ID NO: 104), WTNNWGTY (SEQ ID NO: 105), WYNNWGVT (SEQ ID NO: 106), WYNNFGKT (SEQ ID NO: 189ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION 107), WTNTYHEY (SEQ ID NO: 108), WTNNWGKT (SEQ ID NO: 109), LIITYHEY (SEQ ID NO: 110), WTNNYGKM (SEQ ID NO: 111), WTNNFGKM (SEQ ID NO: 112), WTNNFGVT (SEQ ID NO: 113), WTNTYGKT (SEQ ID NO: 114), WYNTYGKT (SEQ ID NO: 115), WTNTWGVT (SEQ ID NO: 116), LIINWGVT (SEQ ID NO: 117), LIINFGKT (SEQ ID NO: 118), WTNNYHEY (SEQ ID NO: 119), WTNNWGKM (SEQ ID NO: 120), WTNTFGKT (SEQ ID NO: 121), WYNTYHEY (SEQ ID NO: 122), WYNTFGKT (SEQ ID NO: 123), WYNTWGVT (SEQ ID NO: 124), WTNTWGTE (SEQ ID NO: 125), LIITYGKM (SEQ ID NO: 126), WHNTWGTE (SEQ ID NO: 127), WYNNWGTY (SEQ ID NO: 128), WTNNWGTE (SEQ ID NO: 129), WYNTYGVT (SEQ ID NO: 130), WTNTYGVT (SEQ ID NO: 131), WHNNWGVT (SEQ ID NO: 132), WTNNYGKT (SEQ ID NO: 133), WTNNFGTY (SEQ ID NO: 134), WHNNFGKT (SEQ ID NO: 135), WYNNYGKM (SEQ ID NO: 136), WYNNFGVT (SEQ ID NO: 137), WTNAYHRY (SEQ ID NO: 138), WYNNWGKT (SEQ ID NO: 139), WYNNFGKM (SEQ ID NO: 140), WTNNFGEY (SEQ ID NO: 141), LIITFGKT (SEQ ID NO: 142), WHNTYGKM (SEQ ID NO: 143), IAYAYHRY (SEQ ID NO: 144), LIINWGTY (SEQ ID NO: 145), LIITWGVT (SEQ ID NO: 146), WTNNYGVT (SEQ ID NO: 147), WTNTWGTY (SEQ ID NO: 148), WYNTWGTE (SEQ ID NO: 149), or WTNNFGKT (SEQ ID NO: 150).

67. A peptide comprising an amino acid sequence comprising one or a combination of two or more of: KDHGIIVA (SEQ ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200), or a functional variant thereof comprising about 85% sequence identity to a respective one of KDHGIIVA (SEQ 190ATTORNEY DOCKET NO STFD-014-PCT PCT APPLICATION ID NO: 151), AHEWWTHN (SEQ ID NO: 152), DRSWWNVM (SEQ ID NO: 153), VWDWWDFM (SEQ ID NO: 154), AHEGIIVA (SEQ ID NO: 155), KDHWWTHN (SEQ ID NO: 156), EHAWWAHT (SEQ ID NO: 157), NVHWWVHQ (SEQ ID NO: 158), NGVWWGMQ (SEQ ID NO: 159), GPKWWGTN (SEQ ID NO: 160), DRSWWTHN (SEQ ID NO: 161), DRSGIIVA (SEQ ID NO: 162), AHEWWNVM (SEQ ID NO: 163), VWDWWTHN (SEQ ID NO: 164), KDHWWNVM (SEQ ID NO: 165), KDHGWTHN (SEQ ID NO: 166), NVHGIIVA (SEQ ID NO: 167), EHAWWTHN (SEQ ID NO: 168), AHEWIIVA (SEQ ID NO: 169), VWDGIIVA (SEQ ID NO: 170), NVHWWTHN (SEQ ID NO: 171), AHEWWAHT (SEQ ID NO: 172), AHEWWDFM (SEQ ID NO: 173), AHEWWVHQ (SEQ ID NO: 174), EHAGIIVA (SEQ ID NO: 175), KDHWWVHQ (SEQ ID NO: 176), KDHWWDFM (SEQ ID NO: 177), KDHWWAHT (SEQ ID NO: 178), GPKWWTHN (SEQ ID NO: 179), NGVWWTHN (SEQ ID NO: 180), NGVGIIVA (SEQ ID NO: 181), GPKGIIVA (SEQ ID NO: 182), AHEWWGTN (SEQ ID NO: 183), AHEWWGMQ (SEQ ID NO: 184), KDHWWGTN (SEQ ID NO: 185), KDHWIIVA (SEQ ID NO: 186), KDHWWGMQ (SEQ ID NO: 187), GNDWWSIT (SEQ ID NO: 188), AHEWWTHT (SEQ ID NO: 189), DRSWIIVA (SEQ ID NO: 190), PDMWWHHT (SEQ ID NO: 191), KDHGWNVM (SEQ ID NO: 192), EHAWWAHN (SEQ ID NO: 193), DRSWWNVA (SEQ ID NO: 194), DRSWWDFM (SEQ ID NO: 195), AHHGIIVA (SEQ ID NO: 196), VWDWWNVM (SEQ ID NO: 197), RKDWWATQ (SEQ ID NO: 198), DRSWWVHQ (SEQ ID NO: 199), or VWDWIIVA (SEQ ID NO: 200).

68. A peptide comprising an amino acid sequence comprising one or a combination of (1) GTHDKCENPKX1X2X3X4X5X6X7X8QNX9NVVX10TNKELEDIYDESN (SEQ ID NO:202), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO:202, wherein X1X2X3X4X5X6X7X8is one of SEQ ID NOS: 1 through 200 or a functional variant thereof comprising about 85% sequence identity to a respective one of SEQ ID NOS: 1 through 200, and X9and X10are any amino acid, and (2) KEETKEVLKKFKEKVNQFX11EHAFDIINKYGDKEIX12NMMX13MLLWRVX14FRSFRI DANNVELIKFNIRVLDWIMAEADNDLSYFISQ (SEQ ID NO:203), or a functional variant thereof comprising from about 75% sequence identity to SEQ ID NO: 203, wherein X11, X12, X13, and X14are any amino acid. 191

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