EP4637792A2

EP4637792A2 - Cd3 and itcr expressing engineered natural killer cells with enhanced function for adoptive immunotherapy

Info

Publication number: EP4637792A2
Application number: EP23908418.9A
Authority: EP
Inventors: Katy REZVANI; Rafet BASAR; Bin Liu; David MARIN COSTA
Original assignee: University of Texas System; University of Texas at Austin
Current assignee: University of Texas System; University of Texas at Austin
Priority date: 2022-12-21
Filing date: 2023-12-20
Publication date: 2025-10-29
Also published as: WO2024137789A3; WO2024137789A2; MX2025007414A; JP2025542308A; CN120548186A; AU2023409234A1; KR20250128342A; IL321219A

Abstract

Embodiments of the disclosure include methods and compositions in which NK cells are modified by the hand of man to express the invariant NK T-cell receptor and CD3 co-receptor on NK cells that do not naturally express them. Such modified NK cells work effectively with bispecific or multi-specific antibodies that are tailored to comprise anti-CD3 antibodies that bind the modified NK cells, thereby triggering signaling, activation, and cytotoxicity of target cells to which the antibodies also bind. Thus, the NK cells are specifically configured to be able to work effectively with Bispecific NK cell engagers (BiKEs) as well as Bispecific T cell Engagers (BiTEs).

Description

CD3 AND ITCR EXPRESSING ENGINEERED NATURAL KILLER CELLS WITH ENHANCED FUNCTION FOR ADOPTIVE IMMUNOTHERAPY

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of U. S. Provisional Patent Application No. 63/434,395 filed December 21, 2022, which is hereby incorporated by reference in its entirety.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted in ST26 format and is hereby incorporated by reference in its entirety. Said ST26 copy, created on December 7, 2023, is named MDAC_P1350WO_Sequence_Listing.xml and is 487,676 bytes in size.

I. Technical Field

[0003] This disclosure relates at least to the fields of immunology, cell biology, molecular biology, and medicine, including at least cancer medicine.

II. Background

[0004] Natural killer (NK) cells have been studied as potential anti-tumor effectors, yet a number of barriers limit their therapeutic exploitation, mainly related to their lack of antigen specificity. One approach to overcome this is to transduce NK cells with a chimeric antigen receptor (CAR) or an engineered T-cell receptor (TCR) to target a desired antigen. In T cells, one can utilize a bispecific or multi-specific antibody, such as a bispecific T cell engager (BiTE) that binds CD3 on the surface of T cells and that also binds an antigen on the surface of cancer cells. CD3 is composed of four distinct chains, and in mammals, the complex contains a CD3y chain, a CD35 chain, and two CD3s chains. These chains associate with the T-cell receptor (TCR) and the ^-chain (zeta-chain) to generate an activation signal in T lymphocytes. However, NK cells do not naturally express the CD3 receptor complex or TCRs.

[0005] The present disclosure satisfies a long-felt need in the art to improve upon immunotherapies including those that utilize NK cells.

BRIEF SUMMARY

[0006] Embodiments of the disclosure include methods and compositions for treatment of an individual with cancer using adoptive cell therapy. In specific embodiments, the individual is provided a therapeutically effective amount of a bipartite therapy that includes both modified NK cells and antibodies that are capable of being able to bind the NK cells to initiate signaling, activation, and killing of target cells. The disclosure concerns NK cells that have been modified to express multiple proteins that are not naturally expressed in NK cells and that work in conjunction together, including heterologous proteins on the surface of the NK cells that are naturally not present in NK cells.

[0007] In certain embodiments, disclosed herein are engineered NK cell modified to express, a) part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3 b) part or all of a single chain or any combination of invariant NK T cell receptor (iTCR) alpha (iTCRa) chain or beta (iTCR0) chain; and c) a cytokine selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, IL-7, GMCSF, and a combination thereof. In certain embodiments, the NK cells are modified to express part or all of one CD36, two of CD3s, one CD3y, and/or one CD3^. In certain embodiments, any one or more of CD36, CD3s, CD3y, and/or CD3<^ are heterologously linked to one or more intracellular signaling domains. In certain embodiments an intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and a combination thereof. In certain embodiments, an intracellular signaling domain comprises a DAP 10 intracellular signaling domain. In certain embodiments, an intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 115. In certain embodiments, an intracellular signaling domain comprises a CD28 intracellular signaling domain. In certain embodiments, an intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 116. In certain embodiments, an intracellular signaling domain comprises a DAP 10 and CD28 intracellular signaling domain. In certain embodiments, an intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 117. In certain embodiments, an NK cell is modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT- NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122).

[0008] In certain embodiments, an iTCRa chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 293, and the iTCR0 chain comprises an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315. In certain embodiments, an iTCRa chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 293, and the iTCR0 chain comprises an amino acid sequence at least 90% identical to SEQ ID NOs: 301, 307, 311, or 313. [0009] In certain embodiments, NK cells are derived from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or a combination thereof. In certain embodiments, NK cells are primary NK cells, and are not derived from stem cells and/or induced pluripotent stem cells (iPSCs). In certain embodiments, NK cells are complexed to one or more antibodies. In certain embodiments, the one or more antibodies are one or more bispecific or multi-specific antibodies, wherein at least one of the bispecific or multi-specific antibodies comprises an anti- CD3 antibody linked. In certain embodiments, the antibody is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B. In certain embodiments, the antibody is Blinatumomab. In certain embodiments, the NK cell expresses the antibody.

[0010] In certain embodiments, a cytokine is membrane-bound. In certain embodiments, the cytokine is IL-15. In certain embodiments, the NK cell is further modified to express one or more additional heterologous proteins selected from the group consisting of an antigen receptor, a cytokine, a homing receptor, a chemokine receptor, and a combination thereof. In certain embodiments, the NK cells are pre-activated with one or more cytokines. In certain embodiments, the cytokines are IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, or a combination thereof. In certain embodiments, the NK cell further comprises one or more engineered mutations in an endogenous gene. In certain embodiments, the endogenous gene is TGFBR2, CISH, and/or CD38.

[0011] In some embodiments, also provided herein are compositions comprising an engineered NK cell disclosed herein.

[0012] Also provided herein are compositions comprising a complex comprising: a) an engineered NK cell modified to express part or all of the CD3 receptor complex, an iTCRa chain, an iTCRP, and one or more cytokines; and b) a bispecific or multi-specific antibody, wherein the bispecific or multi-specific antibody comprises an anti-CD3 antibody that is bound to CD3 on the NK cells. In some embodiments, the engineered NK is cell modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT- NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122). In some embodiments, the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCRP chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315. In some embodiments, the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCRP chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 301, 307, 311, or 313. In some embodiments, the bispecific or multi-specific antibody is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B. In some embodiments, the bispecific or multi-specific antibody is Blinatumomab. In some embodiments, compositions further comprise a pharmaceutically acceptable excipient. In some embodiments, the composition is comprised in a delivery device.

[0013] Also disclosed herein are methods of treating a disease in an individual. In some embodiments, a method of treating a disease in an individual comprises the step of administering to the individual a therapeutically effective amount of any one of the cells or compositions of claims 1 to 36. In some embodiments, the disease is an autoimmune disease, infection, and/or cancer. In some embodiments, the method comprising the step of administering to the individual an engineered NK cell modified to express a) part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3 b) part or all of a single chain or any combination of invariant NK T cell receptor (iTCR) alpha (iTCRa) chain or beta (iTCRP) chain; and c) a cytokine selected from the group consisting of IL-15, IL-12, IL-2, IL- 18, IL-21, IL-23, IL-7, GMCSF, and a combination thereof. In some embodiments, a method further comprises administering to the individual at the same time or at different time, one or more bispecific or multispecific antibodies. In some embodiments, the one or more bispecific or multispecific antibodies is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B. In some embodiments, the one or more bispecific or multispecific antibodies and the engineered NK cell are administered at the same time, and/or wherein the one or more bispecific or multispecific antibodies and the engineered NK cells are complexed prior to administration to the individual. In some embodiments, the engineered NK is cell modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT-NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122). In some embodiments, the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCRP chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315. In some embodiments, the disease is cancer.

[0014] Certain embodiments of the present invention are characterized through the following enumerated aspects. [0015] Aspect 1, is an engineered NK cell modified to express, a) part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3 b) part or all of a single chain or any combination of invariant NK T cell receptor (iTCR) alpha (iTCRa) chain or beta (iTCR0) chain; and c) a cytokine selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL- 21, IL-23, IL-7, GMCSF, and a combination thereof.

[0016] Aspect 2, is the engineered NK cell of aspect 1, wherein the NK cells are modified to express part or all of CD36, two of CD3s, CD3y, and/or CD3^.

[0017] Aspect 3, is the engineered NK cell of aspect 1 or 2, wherein any one or more of CD36, CD3s, CD3y, and/or CD3(^ are heterologously linked to one or more intracellular signaling domains.

[0018] Aspect 4, is the engineered NK cell of aspect 3, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and a combination thereof.

[0019] Aspect 5, is the engineered NK cell of aspect 3 or 4, wherein the intracellular signaling domain comprises a DAP 10 intracellular signaling domain.

[0020] Aspect 6, is the engineered NK cell of aspect 5, wherein the intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 115.

[0021] Aspect 7, is the engineered NK cell of aspect 3 or 4, wherein the intracellular signaling domain comprises a CD28 intracellular signaling domain.

[0022] Aspect 8, is the engineered NK cell of aspect 7, wherein the intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 116.

[0023] Aspect 9, is the engineered NK cell of aspect 3 or 4, wherein the intracellular signaling domain comprises a DAP 10 and CD28 intracellular signaling domain.

[0024] Aspect 10, is the engineered NK cell of aspect 9, wherein the intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 117.

[0025] Aspect 11, is the engineered NK cell of any one of aspects 1 to 10, wherein the iTCRa chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 293, and the iTCR0 chain comprises an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315.

[0026] Aspect 12, is the engineered NK cell of any one of aspects 1 to 11, wherein the iTCRa chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 293, and the iTCR0 chain comprises an amino acid sequence at least 90% identical to SEQ ID NOs: 301, 307, 311, or 313. [0027] Aspect 13, is the engineered NK cell of any one of aspects 1 to 12, wherein the cell is modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT-NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122).

[0028] Aspect 14, is the engineered NK cell of any one of aspects 1 to 13, wherein the NK cells are derived from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or a combination thereof.

[0029] Aspect 15, is the engineered NK cell of any one of aspects 1 to 14, wherein the NK cells are primary NK cells, and are not derived from stem cells and/or induced pluripotent stem cells (iPSCs).

[0030] Aspect 16, is the engineered NK cell of any one of aspects 1 to 15, wherein the NK cells are complexed to one or more antibodies.

[0031] Aspect 17, is the engineered NK cell of aspect 16, wherein the one or more antibodies are one or more bispecific or multi-specific antibodies, wherein at least one of the bispecific or multi-specific antibodies comprises an anti-CD3 antibody linked.

[0032] Aspect 18, is the engineered NK cell of aspect 16 or 17, wherein the antibody is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B.

[0033] Aspect 19, is the engineered NK cell of any one of aspects 16 to 18, wherein the antibody is Blinatumomab.

[0034] Aspect 20, is the engineered NK cell of any one of aspects 16 to 19, wherein the NK cell expresses the antibody.

[0035] Aspect 21, is the engineered NK cell of any one of aspects 1 to 20, wherein the cytokine is membrane-bound.

[0036] Aspect 22, is the engineered NK cell of any one of aspects 1 to 21, wherein the cytokine is IL-15.

[0037] Aspect 23, is the engineered NK cell of any one of aspects 1 to 22, wherein the NK cell is further modified to express one or more additional heterologous proteins selected from the group consisting of an antigen receptor, a cytokine, a homing receptor, a chemokine receptor, and a combination thereof.

[0038] Aspect 24, is the engineered NK cell of any one of aspects 1 to 23, wherein the NK cells are pre-activated with one or more cytokines.

[0039] Aspect 25, is the engineered NK cell of aspect 24, wherein the cytokines are IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, or a combination thereof. [0040] Aspect 26, is the engineered NK cell of any one of aspects 1 to 25, wherein the NK cell further comprises one or more engineered mutations in an endogenous gene.

[0041] Aspect 27, is the engineered NK cell of aspect 26, wherein the endogenous gene is TGFBR2, CISH, and/or CD38.

[0042] Aspect 28, is a composition comprising the engineered NK cell of any one of aspects 1 to 27.

[0043] Aspect 29, is a composition comprising a complex comprising: a) an engineered NK cell modified to express part or all of the CD3 receptor complex, a iTCRa chain, a iTCRp, and one or more cytokines; and b) a bispecific or multi-specific antibody, wherein the bispecific or multi-specific antibody comprises an anti-CD3 antibody that is bound to CD3 on the NK cells.

[0044] Aspect 30, is the composition of aspect 29, wherein the engineered NK is cell modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT-NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122).

[0045] Aspect 30, is the composition of aspect 29 or 30, wherein the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCR0 chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315.

[0046] Aspect 32, is the composition of any one of aspects 29 to 31, wherein the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCR0 chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 301, 307, 311, or 313.

[0047] Aspect 33, is the composition of any one of aspects 29 to 32, wherein the bispecific or multi-specific antibody is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B.

[0048] Aspect 34, is the composition of any one of aspects 29 to 33, wherein the bispecific or multi-specific antibody is Blinatumomab.

[0049] Aspect 35, is the composition of any one of aspects 29 to 34, further comprising a pharmaceutically acceptable excipient.

[0050] Aspect 36, is the composition of any one of aspects 29 to 35, wherein the composition is comprised in a delivery device. [0051] Aspect 37, is a method of treating a disease in an individual, the method comprising the step of administering to the individual a therapeutically effective amount of any one of the cells or compositions of aspects 1 to 36.

[0052] Aspect 38, is the method of aspect 37, wherein the disease is an autoimmune disease, infection, and/or cancer.

[0053] Aspect 39, is a method of treating a disease in an individual, the method comprising the step of administering to the individual an engineered NK cell modified to express a) part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3 b) part or all of a single chain or any combination of invariant NK T cell receptor (iTCR) alpha (iTCRa) chain or beta (iTCR0) chain; and c) a cytokine selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, IL-7, GMCSF, and a combination thereof.

[0054] Aspect 40, is the method of aspect 39, further comprising administering to the individual at the same time or at different time, one or more bispecific or multispecific antibodies.

[0055] Aspect 41, is the method of aspect 40, wherein the one or more bispecific or multispecific antibodies is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B.

[0056] Aspect 42, is the method of aspect 40 or 41, wherein the one or more bispecific or multispecific antibodies and the engineered NK cell are administered at the same time, and/or wherein the one or more bispecific or multispecific antibodies and the engineered NK cells are complexed prior to administration to the individual.

[0057] Aspect 43, is the method of any one of aspects 39 to 42, wherein the engineered NK is cell modified to express a polynucleotide sequence at least 85% identical to UT-NK15- DAP10 (SEQ ID NO: 118), UT-NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122).

[0058] Aspect 44, is the method of any one of aspects 39 to 43, wherein the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCR0 chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315. [0059] Aspect 45, is the method of any one of aspects 39 to 44, wherein the disease is cancer.

[0060] Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

[0061] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0062] FIG. 1A-1C illustrates various embodiments of NK cells engineered to express CD3, including for use with a variety of heterologous proteins, such as cytokines, bi-specific NK cell engagers, and engineered antigen receptors (CAR and/or TCR). FIG. IB illustrates NK cells accommodated for CD3 and TCR for optimal cancer immunotherapy. FIG. 1C illustrates examples of single chimeric CD3 constructions.

[0063] FIG. 2A-2B illustrates one example of an expression construct for CD3 receptor complex components for transduction or transfection of NK cells. FIG. 2B shows an example of a plasmid map for the representative expression construct.

[0064] FIG. 3 provides a table of various TCR/CD3 expression construct designs for NK- TCR engineering.

[0065] FIG. 4 shows CD3 expression at day 4 on engineered NK cells after transduction with one example of a CMV-directed TCR complex.

[0066] FIG. 5 demonstrates TCR expression at day 4 on engineered NK cells following CMV-directed TCR complex transduction.

[0067] FIG. 6 shows TCR/CD3 expression at day 6 on engineered NK cells after transduction of a CMV-directed TCR complex into the cells.

[0068] FIG. 7 demonstrates binding at different concentrations of one example of a CD3- CD19 BiTE on NK cells through the CD3/TCR complex on the NK cells.

[0069] FIG. 8 shows NK-TCR cytokine production of TNFa and CD107a after stimulation with plate-bound CD3 antibody.

[0070] FIG. 9 demonstrates phosphorylation of CD3z in NK TCR/CD3 cells after crosslinking CD3. [0071] FIGS. 10A-10B show that pre-culturing CD3-CD19 BiTEs with TCR/CD3- expressing NK cells increased its killing activity against Raji cells. FIG. 10A represents a 1: 1 Effector: Target ratio, and FIG. 10B represents a 1 :5 Effector: Target ratio.

[0072] FIG. 11 provides a schematic overview of multiple retroviral transductions to generate NK cells expressing CD3, IL-15, and a TCR complex.

[0073] FIG. 12 shows expression of NY-ESO TCR on NK cells transduced with uTNK15. WT refers to wild type CD3 molecules with IL- 15; A refers to CD3-CD28 with IL- 15; B refers to CD3-DAP10 with IL-15; and C refers to CD3-CD28-DaplO with IL-15.

[0074] FIG. 13 shows the number of TCR molecules per cell expressed on NK cells. WT refers to wild type CD3 molecules with IL- 15; A refers to CD3-CD28 with IL- 15; B refers to CD3-DAP10 with IL- 15; and C refers to CD3-CD28-DaplO with IL-15. Phycoerythrin Fluorescence Quantitation Kit (BD Biosciences) was used to determine the number of molecules of NY-ESO TCR on NK cells.

[0075] FIG. 14 shows expression of NY-ESO TCR on T cells.

[0076] FIG. 15 shows that NK cells transduced with NY-ESO TCR kill NY-ESO peptide- pulsed target cells in a dose-dependent manner. WT refers to wild type CD3 molecules with IL- 15; A refers to CD3-CD28 with IL-15; B refers to CD3-DAP10 with IL-15; and C refers to CD3-CD28-DaplO with IL-15.

[0077] FIG. 16 demonstrates endogenous NY-ESO expression on human tumor cell lines. [0078] FIG. 17 demonstrates that NY-ESO TCR transduced T cells kill NY-ESO expressing tumor targets.

[0079] FIG. 18 provides results that NY-ESO TCR transduced NK cells kill NY-ESO expressing tumor targets even at low E:T ratios. WT refers to wild type CD3 molecules with IL- 15; A refers to CD3-CD28 with IL-15; B refers to CD3-DAP10 with IL-15; and C refers to CD3-CD28-DaplO with IL-15.

[0080] FIGs. 19A-19B show that NY-ESO transduced NK cells have a similar phenotype (FIG. 19A) and expression pattern (FIG. 19B) to NT NK cells. WT refers to wild type CD3 molecules with IL-15; A refers to CD3-CD28 with IL-15; B refers to CD3-DAP10 with IL- 15; and C refers to CD3-CD28-DaplO with IL-15.

[0081] FIG. 20 provides a table representing the cellular composition of the expanded uTNK15 product. WT refers to wild type CD3 molecules with IL- 15; A refers to CD3-CD28 with IL- 15; B refers to CD3-DAP10 with IL- 15; and C refers to CD3-CD28-DaplO with IL- 15. [0082] FIG. 21A-21C shows that NK cells can be successfully transduced with CD3 and TCR constant alpha-beta (TCRCab) (called TCR6 construct) and that the engineered NK cell can bind Blinatumumab (FIG. 21B) and selectively kill CD 19+ lymphoma targets (FIG. 21C). [0083] FIGs. 22A-22C shows the in vivo activity of effector cells (e.g., NK cells, or T cells) comprising NY-ESO targeted TCRs. FIG. 22A is a schematic outlining the experimental procedure performed. FIG. 22B displays bioluminescent imaging over time (day 1, day 7, day 14, and day 21) for the mice engrafted with U266B.1 cells transduced with FireFlyluciferase (FFluc) and treated with control, NY-ESO TCR NK cells, or NY-ESO TCR T cells (NK cells comprising WT, #A, or #B UT-NK15-NY ESO TCR constructs respectively; WT refers to wild type CD3 molecules with IL-15; #A refers to CD3-CD28 with IL-15; and #B refers to CD3- DAP10 with IL- 15). FIG. 22C is a graphical quantification of the bioluminescence average radiance displayed in FIG 22B. These results showed that effector cells comprising NY-ESO TCR constructs described herein robustly inhibited tumor growth in vivo.

[0084] FIGs. 23A-23B shows the in vitro activity of effector cells (e.g., NK cells or T cells) comprising NY-ESO targeted TCRs and UT-NK15 constructs. FIG. 23A are images of spheroids formed by osteosarcoma tumor cell line Saos-2 stably transduced to express GFP that were used to test the activity of NY-ESO 1 -specific TCR expressing NK and T cells cytotoxicity. FIG. 23B is a graph showing percentage of cytotoxicity (Y axis) for representative images after 3 days of co-culture. NK cells were co-transduced with NY-ESO- TCR, and the UT-NK15 signaling complex co-expressing different co-stimulatory molecules fused to the CD3(^ signaling chain or the TCR complex without IL-15. T cells were only transduced with NY-ESO TCR. Abbreviation in the graph: 28 = CD3(^ fused to a CD28 costimulatory domain; 10 = CD3(^ fused to a Dap 10 co-stimulatory domain; 8 = CD8 alpha/beta co-receptor as part of the NY ESO TCR construct; wo IL- 15 = the construct only contains CD3 zeta, epsilon, gamma and delta TCR complex without co-stimulation or IL-15.

[0085] FIGs. 24A-24D shows the in vivo activity of effector cells (e.g., NK cells or T cells) comprising NY-ESO targeted TCRs and UT-NK15 constructs. FIG. 24A depicts a plan for an in vivo study to test the activity of different NY ESO TCR transduced NK and T cells. FIG. 24B depicts BLI imaging results of the test outlined and performed according to FIG. 24A, Mice were injected with U266 tumor cells, and three days later received T cells transduced with NY-ESO-specific TCR, or NK cells co-transduced with NY-ESO TCR and UT-NK15 with CD3< fused to CD28 (labelled as NY-ESO NK UT-NK15 CD28 or NY-ESO TCR UTNK- 15 CD28 NK cells). The tumor alone group was used as control. FIG. 24C depicts region of interest average radiance intensity for the animals tested according to FIG. 24A and imaged in FIG. 24B. FIG. 24D is a graph depicting the cohort survival curves for the aforementioned animals.

[0086] FIG. 25 shows the in vivo activity of effector cells (e.g., NK cells) engineered to express NY ESO TCR and CD3 complex with or without IL-15 transgene comprised in the construct. NSG mice were irradiated (300 cGy) and the next day were injected with 500,000 U266 cells (HLA-A2 positive, NY-ESO-expressing myeloma cell line) via the tail vein. Three days later, mice received 5 million TCR transduced T or NK cells. Mice were monitored for tumor control by BLI imaging. NK cells were transduced with NY-ESO-specific TCR with or without expression of CD8 alpha/beta co-receptors, co-transduced with CD3 complex without IL-15 transgene or with UT-NK15 expressing CD3(^ fused to CD28 (UT-NK15 CD28) or CD3(^ fused to DAP 10 (UT-NK15 DAP 10) co-stimulatory molecules.

[0087] FIGs. 26A-26C shows in vitro expression of Preferentially Expressed Antigen in Melanoma (PRAME) TCRs on effector cells (e.g., NK cells or T cells) and the in vitro activity of said cells. FIG. 26A shows the expression of both UT-NK15 (x-axis, CD3) and PRAME- specific TCRs (y-axis, TCR) in NK cells (TCR clones 46, 54, or DSK3 respectively), or the expression of PRAME-specific TCRs in T cells transduced with the same (TCR clones 46 or 54). FIG. 26B shows the in vitro cytotoxicity of NK cells expressing a PRAME-specific TCR against the U266 myeloma cell line. IncuCyte® live cell imaging was used to measure the cytotoxicity of T cells transduced with PRAME-specific TCR and NK cells transduced with UT-NK15 and PRAME-specific TCR against U266 myeloma cells. GFP-expressing U266 cells were co-cultured with PRAME-specific TCR expressing T cell or NK cells at 1 : 1 effector : target ratio. A reduction in GFP expression indicated cell death. After 26 hours, a second round of 50,000 tumor cells was added (noted as “rechallenging”) to each well for the tumor rechallenge assay. Open symbols represent T cells, while closed symbols represent NK cells. NT = non-transduced. FIG. 26C shows the in vitro cytotoxicity of NK cells expressing a PRAME-specific TCR against the UA375 melanoma cell line. IncuCyte® live cell imaging was used to measure the cytotoxicity of T cells transduced with PRAME-specific TCR and NK cells transduced with UT-NK15 and PRAME-specific TCR (PRAME-specific TCR clone 46 (TCR-46), PRAME-specific TCR clone 54 (TCR-54), or PRAME-specific TCR clone DSK3 (DSK)) against UA375 melanoma cells. GFP-expressing UA375 cells were co-cultured with PRAME-expressing T cell or NK cells at 1 : 1 effector : target ratio. A reduction in GFP expression indicated cell death. After 26 hours, a second round of 50,000 tumor cells was added to each well for the tumor rechallenge assay. Open symbols represent T cells, while closed symbols represent NK cells. NT = non-transduced. [0088] FIGs. 27A-27D shows in vitro expression of, and the cytotoxic function of, invariant Natural Killer T-cell Receptors (iTCRs) in NK cells. Additional experimental details are provided in Example 6 below. FIG. 27A depicts exemplary flow cytometry plots showing expression of three different iTCR pairs, each comprising iTCRa (SEQ ID NO: 295), and either iTCRp chain 1 (SEQ ID NO: 297), iTCRp chain 2 (SEQ ID NO: 299), or iTCRp chain 3 (SEQ ID NO: 301), and CD3 on NK cells, using antibodies specific for iTCR target Va24 and Vpi 1 regions (y and x axis respectively). FIG. 27B depicts exemplary flow cytometry plots showing binding of Blinatumomab (y axis) to three different CD3/iTCR complexes as described in (A), e.g., iTCRl, iTCR2, and iTCR3, respectively, expressing NK cells. NK cells were derived from three donors, cell donor 4 (CD4), cell donor 8 (CD 8), or cell donor N (CDN), respectively. The binding of Blinatumomab on CD3 was confirmed by flow cytometry using the CAR19 detection kit from Miltenyi Biotech™. FIG. 27C depicts results of IncuCyte® live cell imaging assays that were used to measure the cytotoxicity of NK cells pre-loaded with Blinatumomab and co-transduced with iTCR and UT-NK15, against GFP-expressing Raji tumor cells at a 3: 1 effector to target ratio. T cells and iNKT cells were used as positive and negative controls, respectively. A reduction in GFP expression indicated cell death. Three NK donor lines were utilized, CB152, CB153, and CB154, respectively. NK cells were transduced with three different CD3/iTCR complexes as described in (A), e.g., iTCRl, iTCR2, or iTCR3, or left nontransduced (NT) (as negative control). FIG. 27D depicts a bar chart showing residual tumor % (Y axis) for the data presented in FIG. 27C. Unpaired t test was utilized to compare iNKT with each of the different iTCRs individually. CD3/iTCR NK cells were significantly (P = <0.01) more cytotoxic than iNKT cells (N = 3 replicates).

[0089] FIGs. 28A-28D shows the cloning of iTCR sequences from NK T Cells isolated from human cord blood, and the transgenic expression and efficacy of a subset of iTCR clones in transduced cord blood derived NK cells. Additional experimental details are provided in Example 6 below. FIG. 28A depicts the isolation of iTCR clones from iNKT cells purified from five cord blood donors, and exemplary flow cytometry results showing expression of iTCRs on said donor iNKT cells. iNKT cells were isolated from 5 cord blood donors using iNKT isolation kit from Miltenyi Biotech™. Isolated cells were stimulated and expanded with irradiated cord blood PBMC(40 Gry) in the presence of 100 ng/ml of alpha-Galactosylceramide and 200 U/ml of IL-2. On day 7 after iNKT expansion, the purity of iNKT cultures was confirmed by iNKT specific antibodies against Va24 and Vpi l. Total mRNA was extracted from iNKT cells, and cDNAs of the VP-DJ regions were cloned and sequenced (100 individual clones). FIG. 28B depicts sequences of iTCRp clone VP-DJ sequences. FIG. 28C depicts the transgenic expression of Va24 and CD3 in transgenic CD3/iTCR expressing NK cell populations from three cord blood donors. The NK cells were co-transduced with iTCRs (eight randomly selected iTCRP clones with coding sequences represented by SEQ ID NOs: 300, 302, 304, 306, 308, 310, 312, or 314, representing clones 3, 18, 24, 51, 56, 76, 93, and 96 respectively, and iTCRa coding sequence represented by SEQ ID NO: 292) and UT-NK15 on day 5. iTCR expression was confirmed by flow cytometry using iTCR specific antibodies against Va24 and antibodies against CD3. FIG. 28D depicts a graph describing the results of IncuCyte® live cell imaging mediated measurements of cytotoxicity of NK cells co-transduced with iTCRs (as described in FIG. 28B) and UT-NK15, and pre-loaded with Blinatumomab (1 hour at 37 °C), against GFP expressing Raji cells at 3 : 1 target effector ratios. T cells were used as positive control and non-transduced (NT) cells were utilized as negative controls. A reduction in GFP expression indicated tumor cell death.

DETAILED DESCRIPTION

[0090] In keeping with long-standing patent law convention, the words “a” and “an” when used in the present specification in concert with the word comprising, including the claims, denote “one or more.” Some embodiments of the disclosure may consist of or consist essentially of one or more elements, method steps, and/or methods of the disclosure. It is contemplated that any method or composition described herein can be implemented with respect to any other method or composition described herein and that different embodiments may be combined.

[0091] Throughout this specification, unless the context requires otherwise, the words “comprise”, “comprises” and “comprising” will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements. By “consisting of’ is meant including, and limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of’ indicates that the listed elements are required or mandatory, and that no other elements may be present. By “consisting essentially of’ is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase “consisting essentially of’ indicates that the listed elements are required or mandatory, but that no other elements are optional and may or may not be present depending upon whether or not they affect the activity or action of the listed elements. [0092] Reference throughout this specification to “one embodiment,” “an embodiment,” “a particular embodiment,” “a related embodiment,” “a certain embodiment,” “an additional embodiment,” or “a further embodiment” or combinations thereof means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the appearances of the foregoing phrases in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0093] As used herein, the terms “or” and “and/or” are utilized to describe multiple components in combination or exclusive of one another. For example, “x, y, and/or z” can refer to “x” alone, “y” alone, “z” alone, “x, y, and z,” “(x and y) or z,” “x or (y and z),” or “x or y or z.” It is specifically contemplated that x, y, or z may be specifically excluded from an embodiment.

[0094] Throughout this application, the term “about” is used according to its plain and ordinary meaning in the area of cell and molecular biology to indicate that a value includes the standard deviation of error for the device or method being employed to determine the value. [0095] As used herein, the term “CD3 receptor complex” or “CD3 co-receptor complex” refers to the protein complex that in nature acts as a T cell co-receptor and is comprised of CD3<^ chain, CD3y chain, a CD35 chain, and two CD3s chains (although in alternatives only one CD3s chain is used).

[0096] The term “engineered” as used herein refers to an entity that is generated by the hand of man, including a cell, nucleic acid, polypeptide, vector, and so forth. In at least some cases, an engineered entity is synthetic and comprises elements that are not naturally present or configured in the manner in which it is utilized in the disclosure. In specific embodiments, a vector is engineered through recombinant nucleic acid technologies, and a cell is engineered through transfection or transduction of an engineered vector. Cells may be engineered to express heterologous proteins that are not naturally expressed by the cells, either because the heterologous proteins are recombinant or synthetic or because the cells do not naturally express the proteins.

[0097] The phrases “pharmaceutical or pharmacologically acceptable” refers to molecular entities and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal, such as a human, as appropriate. The preparation of a pharmaceutical composition comprising an antibody or additional active ingredient will be known to those of skill in the art in light of the present disclosure. Moreover, for animal (e.g., human) administration, it will be understood that preparations should meet sterility, pyrogenicity, general safety, and purity standards as required by FDA Office of Biological Standards.

[0098] As used herein, “pharmaceutically acceptable carrier” includes any and all aqueous solvents (e.g., water, alcoholic/aqueous solutions, saline solutions, parenteral vehicles, such as sodium chloride, Ringer's dextrose, etc.), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oil, and injectable organic esters, such as ethyloleate), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, anti-oxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, fluid and nutrient replenishers, such like materials and combinations thereof, as would be known to one of ordinary skill in the art. The pH and exact concentration of the various components in a pharmaceutical composition are adjusted according to well-known parameters.

[0099] The term “subject,” as used herein, generally refers to an individual having or is suspected of having cancer. The subject can be any organism or animal subject that is an object of a method or material, including mammals, e.g., humans, laboratory animals (e.g., primates, rats, mice, rabbits), livestock (e.g., cows, sheep, goats, pigs, turkeys, and chickens), household pets (e.g., dogs, cats, and rodents), horses, and transgenic non-human animals. The subject can be a patient, e.g., have or be suspected of having a disease (that may be referred to as a medical condition), such as benign or malignant neoplasias, or cancer. The subject may be undergoing or having undergone treatment. The subject may be asymptomatic. The subject may be healthy individuals but that are desirous of prevention of cancer. The term “individual” may be used interchangeably, in at least some cases. The “subject” or "individual", as used herein, may or may not be housed in a medical facility and may be treated as an outpatient of a medical facility. The individual may be receiving one or more medical compositions via the internet. An individual may comprise any age of a human or non-human animal and therefore includes both adult and juveniles (i.e., children) and infants and includes in utero individuals. It is not intended that the term connote a need for medical treatment, therefore, an individual may voluntarily or involuntarily be part of experimentation whether clinical or in support of basic science studies.

[0100] As used herein “treatment” or “treating,” includes any beneficial or desirable effect on the symptoms or pathology of a disease or pathological condition, and may include even minimal reductions in one or more measurable markers of the disease or condition being treated, e.g., cancer. Treatment can involve optionally either the reduction or amelioration of one or more symptoms of the disease or condition, or the delaying of the progression of the disease or condition. “Treatment” does not necessarily indicate complete eradication or cure of the disease or condition, or associated symptoms thereof. Treating may mean alleviation of at least one symptom of the disease or condition.

[0101] As used herein “TCR/CD3 complex” refers to a protein complex naturally found on the surface of T cells and that comprises T-cell receptor (TCR) a and 0 chains, invariant Natural Killer T-cell receptor (iTCR) a and 0 chains, and/or a T-cell receptor y and 5 chains, in addition to CD3^, CD3y, CD36, and CD3s chains.

I. Embodiments of the Disclosure

[0102] Natural killer (NK) cells are an emerging cellular immunotherapy for patients with malignant hematologic disease, as well as solid tumors. The present disclosure specifically relates to NK cells that have been modified to render the NK cells to have enhanced function as an immunotherapy compared to NK cells not so modified. The modifications allow for the NK cells to have greater versatility when used with other therapeutic agents and at least in some embodiments to have T cell-like activity by utilizing the CD3/TCR receptor complex. In specific embodiments, the NK cells are modified to express (i) either a single CD3 chain (CD3zeta, CD3 epsilon, CD3 delta, or CD3 gamma) or part or all of the human CD3 receptor complex (including any combination of CD3 delta, epsilon (one or two copies of epsilon), gamma, and zeta); or (ii) either a single CD3 chain or the human CD3 receptor complex (including any combination of CD3 delta, epsilon (one or two molecules), gamma, and zeta) as a full length protein or as a partial protein heterologously linked to one or more intracellular signaling domains); and (iii) the CD3 complex may or may not include the T-cell receptor (a0 or y6) and/or iTCR receptor (a0). The disclosure concerns the use of CD3 -expressing NK cells in the diagnosis and treatment of disease, including use of the cells in combination with bispecific or multi-specific antibodies in which one epitope of the antibody binds CD3 on the CD3 -expressing NK cells. The CD3 -expressing NK cells can either be pre-complexed ex vivo with the bi/multi-specific antibody to redirect their specificity toward the target antigen and/or combined in vivo. In diagnostic embodiments, labeled NK cells may be loaded with bispecific or multi-specific antibodies of any kind, including that comprise at least an anti-CD3 antibody, and the loaded, labeled NK cells may be monitored for trafficking to the site of the target antigen for which another antibody on the bispecific or multi-specific antibody binds. [0103] In certain embodiments of the disclosure, a TCR recognizes antigens and/or epitopes presented by a major-histocompatibility complex (MHC). In certain embodiments, an antigen and/or epitope is a peptide, lipid, and/or glycolipid. In certain embodiments, a MHC is a class I MHC. In certain embodiments, a MHC is a class II MHC. In certain embodiments, an MHC is a non-classical MHC. In certain embodiments, an MHC is a class I-like MHC. In certain embodiments, an MHC is CD Id.

[0104] In certain embodiments, a TCR target antigen is not primarily what provides a transduced effector cell with target antigen specificity. In certain embodiments, a TCR acts primarily as a stabilizer for a CD3 co-receptor complex, while an antibody provides the primary target antigen specificity for a transduced effector cell.

IL Compositions of the Disclosure

[0105] The disclosure concerns compositions that at least include modified NK cells that express at least parts of the TCR/CD3 complex. In some cases, the compositions also include bispecific or multi-specific antibodies, including in the same formulation, although in alternative embodiments the NK cells and antibodies are utilized as physically separate compositions.

A. NK Cell TCR/CD3 Modifications

[0106] In particular embodiments, provided herein are compositions that comprise NK cells that have been modified by the hand of man to express part or all of the TCR receptor complex, iTCR receptor complex, and/or part or all of the CD3 co-receptor complex. In specific embodiments, the NK cells are modified to include all components of the CD3 complex, including CD3<^, CD3s, CD3y and CD36. Although in particular cases the full lengths of CD3<^, CD3s, CD3y and CD36 are utilized, including their extracellular domain, transmembrane domain, and intracellular domain, in alternative embodiments only part of one or more of CD3<^, CD3s, CD3y and CD36 are utilized each of which that may or may not be combined with one or more intracellular signaling domains such as CD 16, NKG2D, DAP 10, DAP 12, CD28, 4 IBB, 2B4, CD27, 0X40, or any combination thereof. The NK cells may also be modified to express the TCR receptor complex and/or iTCR receptor complex, although in alternative embodiments none of the TCR receptor complex or iTCR receptor complex components are utilized.

[0107] In certain embodiments, an amino acid sequence (e.g., a polypeptide) may comprise an amino acid represented by a single letter “X” or a three letter code “Xaa”. In some embodiments, the amino acid represented by “X” or “Xaa” is any naturally occurring amino acid, such as but not limited to, Arginine (Arg, R), Histidine (His, H), Lysine (Lys, K), Aspartic Acid (Asp, D), Glutamic Acid (Glu, E), Serine (Ser, S), Threonine (Thr, T), Asparagine (Asn, N), Glutamine (Gin, Q), Glycine (Gly, G), Proline (Pro, P), Cysteine (Cys, C), Alanine (Ala, A), Valine (Vai, V), Isoleucine (He, I), Leucine (Leu, L), Methionine (Met, M), Phenylalanine (Phe, F), Tyrosine (Tyr, Y), or Tryptophan (Trp, W).

[0108] In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Arginine (Arg, R). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Histidine (His, H). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Lysine (Lys, K). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Aspartic Acid (Asp, D). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Glutamic Acid (Glu, E). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Serine (Ser, S). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Threonine (Thr, T). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Asparagine (Asn, N). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Glutamine (Gin, Q). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Glycine (Gly, G). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Proline (Pro, P). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Cysteine (Cys, C). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Alanine (Ala, A). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Valine (Vai, V). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Isoleucine (He, I). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Leucine (Leu, L). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 in SEQ ID NO: 25 or SEQ ID NO: 88 is Methionine (Met, M). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Phenylalanine (Phe, F). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Tyrosine (Tyr, Y). In some embodiments, the amino acid represented by “X” or “Xaa” in SEQ ID NO: 25 or SEQ ID NO: 88 is Tryptophan (Trp, W).

[0109] In certain embodiments, particular sequences for any of the CD3 receptor components are utilized, including wildtype or mutants of the components so long as the CD3 receptor having the mutant is able to allow signaling through the CD3 complex leading to activation and killing of targets. In some cases, the following examples of sequences for CD3s, CD36, CD3y, and CD3^ and are utilized for modification of the NK cells.

[0110] CD3 Epsilon (UniProtKB - P07766 (CD3E HUMAN))

[0111] Signal Peptide

MQSGTHWRVLGLCLLSVGVW ( SEQ ID NO : 1 )

[0112] Extracellular Domain sp|P07766|23-126

DGNEEMGGI TQTPYKVS I SGTTVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDHLSL KEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD ( SEQ ID NO : 2 )

[0113] Transmembrane Domain sp|P07766| 127- 152

VMSVAT IVIVDICI TGGLLLLVYYWS ( SEQ ID NO : 3 )

[0114] Intracellular Domain sp|P07766| 153-207

KNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI ( SEQ ID NO : 4 )

[0115] An example of a Homo sapiens CD3e molecule (CD3E), mRNA is at NCBI Reference Sequence: GENBANK® Accession No. NM_000733.4

ATGCAGTCGGGCACTCACTGGAGAGTTCTGGGCCTCTGCCTCTTATCAGTTGGCGTTTGGGG GCAAGATGGTAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCTCTG GAAC GAG AG T AAT AT T GAG AT G C C C T GAG TATCCTGGATCT GAAAT AC T AT G G C AAC AC AAT GATAAAAACATAGGCGGTGATGAGGATGATAAAAACATAGGCAGTGATGAGGATCACCTGTC ACTGAAGGAATTTTCAGAATTGGAGCAAAGTGGTTATTATGTCTGCTACCCCAGAGGAAGCA AACCAGAAGATGCGAACTTTTATCTCTACCTGAGGGCAAGAGTGTGTGAGAACTGCATGGAG ATGGATGTGATGTCGGTGGCCACAATTGTCATAGTGGACATCTGCATCACTGGGGGCTTGCT GCTGCTGGTTTACTACTGGAGCAAGAATAGAAAGGCCAAGGCCAAGCCTGTGACACGAGGAG CGGGTGCTGGCGGCAGGCAAAGGGGACAAAACAAGGAGAGGCCACCACCTGTTCCCAACCCA GACTATGAGCCCATCCGGAAAGGCCAGCGGGACCTGTATTCTGGCCTGAATCAGAGACGCAT CTGA ( SEQ ID NO : 5 )

[0116] Examples of respective nucleic acid and amino acid CD3 epsilon sequences in their entirety are as follows (underlining refers to signal peptide sequence): ATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGGG CCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGCG GCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAAC GACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGAG CCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGCA AGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGAG ATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGCT GCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGCG CCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCCC GACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAAT C ( SEQ ID NO : 37 )

MQSGTHWRVLGLCLLSVGVWGQDGNE EMGG I T Q T P YKVS I S G T T V I L T C P Q Y P G S E I L WQHN DKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCME MDVMSVAT IVIVDICI TGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNP DYEPIRKGQRDLYSGLNQRRI ( SEQ ID NO : 38 )

[0117] CD3 Delta (UniProtKB - P04234 (CD3D HUMAN))

[0118] Signal Peptide

MEHSTFLSGLVLATLLSQVS ( SEQ ID NO : 6 )

[0119] Extracellular Domain sp|P04234|22-105

FKI PIEELEDRVFVNCNTS I TWVEGTVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKE STVQVHYRMCQSCVELDPATVA ( SEQ ID NO : 7 )

[0120] Transmembrane Domain sp|P04234| 106-126

GI IVTDVIATLLLALGVFCFA ( SEQ ID NO : 8 )

[0121] Intracellular Domain sp|P04234|127-171

GHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK ( SEQ ID NO : 9 )

[0122] Homo sapiens CD3d molecule, delta (CD3-TCR complex), mRNA (cDNA clone MGC:88324 IMAGE:30412345), complete cds GENBANK®: BC070321.1

ATGGAACATAGCACGTTTCTCTCTGGCCTGGTACTGGCTACCCTTCTCTCGCAAGTGAGCCC C T T C AAGAT AC C T AT AGAG GAAC T T GAG GAG AGAG T G T T T G T GAAT T G C AAT AC C AG CAT C A CATGGGTAGAGGGAACGGTGGGAACACTGCTCTCAGACATTACAAGACTGGACCTGGGAAAA CGCATCCTGGACCCACGAGGAATATATAGGTGTAATGGGACAGATATATACAAGGACAAAGA ATCTACCGTGCAAGTTCATTATCGAATGTGCCAGAGCTGTGTGGAGCTGGATCCAGCCACCG TGGCTGGCATCATTGTCACTGATGTCATTGCCACTCTGCTCCTTGCTTTGGGAGTCTTCTGC TTTGCTGGACATGAGACTGGAAGGCTGTCTGGGGCTGCCGACACACAAGCTCTGTTGAGGAA TGACCAGGTCTATCAGCCCCTCCGAGATCGAGATGATGCTCAGTACAGCCACCTTGGAGGAA ACTGGGCTCGGAACAAGTGA ( SEQ ID NO : 10 ) [0123] Examples of respective nucleic acid and amino acid CD3 delta sequences in their entirety are as follows (underlining refers to signal peptide sequence):

ATGGAGCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCC CTTCAAGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCA CCTGGGTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAG AGAATCCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGA GAGCACCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCG TGGCCGGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGC TTCGCCGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAA CGACCAGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCA ACTGGGCCAGAAACAAG ( SEQ ID NO : 35 )

MEHSTFLSGLVLATLLSQVSPFKI PIEELEDRVFVNCNTS I TWVEGTVGTLLSDI TRLDLGK RILDPRGI YRCNGTDI YKDKESTVQVHYRMCQSCVELDPATVAGI IVTDVIATLLLALGVFC FAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK ( SEQ ID NO : 36 )

[0124] CD3 Gamma (T-cell surface glycoprotein CD3 gamma chain Gene CD3G P09693) Signal Peptide

MEQGKGLAVLILAI ILLQGTLA ( SEQ ID NO : 11 )

[0125] Extracellular Domain sp|P09693|23-l 16

QS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKMIGFLTEDKKKWNLGSNAKDPRGM YQCKGSQNKSKPLQVYYRMCQNCIELNAAT I S ( SEQ ID NO : 12 )

[0126] Transmembrane Domain sp|P09693|l 17-137

GFLFAE IVS I FVLAVGVYFIA ( SEQ ID NO : 13 )

[0127] Intracellular Domain sp|P096931138- 182

GQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRN ( SEQ ID NO : 14 )

[0128] Homo sapiens CD3g molecule (CD3G), mRNA; NM_000073.3:81-629 Homo sapiens CD3g molecule (CD3G), mRNA

ATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTT GGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGG TAG T T C T GAG T T GT GAT GCAGAAGCCAAAAATAT CACAT GGT T TAAAGAT GGGAAGAT GAT C GGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGAGG GAT G TAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T GTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTC AGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTC GAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATC GAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATTGA ( SEQ ID NO : 15 ) [0129] Examples of respective nucleic acid and amino acid CD3 gamma sequences in their entirety are as follows (underlining refers to signal peptide sequence):

ATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTT GGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGG TAG T T C T GAG T T GT GAT GCAGAAGCCAAAAATAT CACAT GGT T TAAAGAT GGGAAGAT GAT C GGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGG GAT G TAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T GTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTC AGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTC GAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATC GAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAAT ( SEQ ID NO : 33 )

MEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKMI GFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAAT I SGFLFAE IV S I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRN ( SEQ ID NO : 34 )

[0130] CD3 Zeta

[0131] Signal Peptide sp|P20963| SP

MKWKALFTAAILQAQLPI TEA ( SEQ ID NO : 1 6 )

[0132] Extracellular Domain sp|P20963122-30 ECD

QS FGLLDPK ( SEQ ID NO : 17 )

[0133] Transmembrane Domain sp|P20963131-51 tmd

LCYLLDGILFI YGVILTALEL ( SEQ ID NO : 18 )

[0134] Intracellular Domain sp|P20963|52-164 ICD

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNE LQKDKMAEAYSE IGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO : 19 )

[0135] Examples of respective nucleic acid and amino acid CD3 zeta sequences in their entirety are as follows (underlining refers to signal peptide sequence):

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCC GCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTA CGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGA AGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGT GAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT CAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGC ( SEQ ID NO : 31 )

MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAP AYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYS EIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO : 32 )

[0136] Homo sapiens CD247 molecule (CD247; also referred to as CD3 Zeta), transcript variant 1, mRNA

NCBI Reference Sequence: NM_198053.3

NM_198053.3:65-559 Homo sapiens CD247 molecule (CD247), transcript variant 1, mRNA ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCC GCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTA CGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGA AGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGT GAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT CAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCTAA ( SEQ ID NO : 20 )

[0137] In specific embodiments, the NK cells are modified to express one of more of the TCRa chain, the TCR0 chain, the TCRy chain, and the TCR5 chain, and any combination thereof may be utilized. In certain embodiments, the TCR may be an invariant Natural Killer cell TCR (iTCR). In a specific case, the NK cells are modified to express the T-cell receptor (TCR) 0.0 chains, iTCR a0 chains, or the TCR y5 chains. In certain cases, the NK cells are modified to express part or all of only the constant region of one of more of the TCRa chain, iTCRa chain, the TCR0 chain, iTCR0 chain, the TCRy chain, and the TCR5 chain. The NK cells may be modified to express part or all of only the constant region of the T-cell receptor (TCR) 0 chains, or the TCR y5 chains, or the iTCR 0 chains. In cases wherein part of the constant region is utilized, the part of the constant region may be at least 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, or 400 amino acids, including contiguous amino acids of any constant region. The part of the constant region may comprise at least 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amino acids of a constant region, including contiguous amino acids of a constant region.

[0138] In specific cases, any sequences encompassed herein are utilized to modify the NK cells, although in other cases sequences that are related to these in identity are utilized. For example, related sequences that are at least 80, 85, 90, 95, 96, 97, 98, 99% identical to any sequence encompassed herein may be utilized in the disclosure. [0139] Particular constructs for the expression of various TCR/CD3 proteins in the NK cells may be utilized, and in a variety of configurations. In specific cases, the NK cells may be transduced or transfected with one or more vectors to express any of the various proteins encompassed herein, including at least any one or more components of the TCR/CD3 complex. In specific cases, the one or more vectors themselves may or may not be multi ci str onic by being able ultimately to produce more than one separate polypeptide. In cases wherein one or more multi ci str onic vectors are employed, they may utilize one or more internal ribosome entry sites (IRES) and/or one or more 2A self-cleaving peptide sites. In cases wherein one or more 2A sequences are utilized, the following may be used, where GSG is an optional linker: [0140] T2A ( GSG) EGRGSLLTCGDVEENPGP ( SEQ ID NO : 21 )

[0141] P2A ( GSG) ATNFSLLKQAGDVEENPGP ( SEQ ID NO : 22 )

[0142] E2A ( GSG) QCTNYALLKLAGDVESNPGP ( SEQ ID NO : 23 )

[0143] F2A ( GSG) VKQTLNFDLLKLAGDVESNPGP ( SEQ ID NO : 24 )

[0144] In situations wherein multiple protein components are expressed from a multi ci str onic vector, the order in a 5' to 3' direction on the polynucleotide vector may be of any order, although in alternative cases they are present on the vector in a particular order. A multi ci str onic vector may express multiple components of the CD3 receptor complex and no other heterologous protein, or the multi ci str onic vector may express multiple components of the CD3 receptor complex and one or more other heterologous proteins. A multi ci str onic vector may express multiple components of the TCR receptor complex and no other heterologous protein, or the multi ci str onic vector may express multiple components of the TCR receptor complex and one or more other heterologous proteins. A multi ci str onic vector may or may not express one or more multiple components of the TCR receptor complex and one or more multiple components of the CD3 complex. In a specific embodiment, a multi ci str onic vector includes one or multiple components of the CD3 receptor complex and one or more heterologous proteins, such as a cytokine and an engineered antigen receptor, such as a CAR. [0145] There is an example in FIG. 2A of a multi ci stronic vector in which full lengths of CD3s, CD36, CD3y, and CD3^ are present and separated by the same or different 2A selfcleaving peptide sites. As further noted in the plasmid map of FIG. 2B, a multicistronic vector may include the signal peptide, extracellular domain, transmembrane domain, and intracellular domain of each of CD3s, CD36, CD3y, and CD3^.

[0146] FIG. 3 provides a table showing examples of various TCR expression constructs for engineering of TCR-expressing NK cells. In particular embodiments of the disclosure, CD3 receptor components and TCR receptor components are expressed from different vectors in the NK cells. In any case, the vector(s) may express a TCR directed against a particular antigen, such as a cancer antigen or a viral antigen. The TCR may or may not comprise at least part of CD3^, including the intracellular domain of CD3^, in addition to the NK cells also expressing CD3^ as a separate molecule from the TCR and as part of the CD3 receptor complex. Likewise, a CAR may or may not comprise at least part of CD3^, including the intracellular domain of CD3^, in addition to the NK cells also expressing CD3^ as a separate molecule from the TCR and as part of the CD3 receptor complex.

[0147] In specific embodiments, a TCR of the modified NK cells is utilized not necessarily as a therapeutic aspect for the cells but as a structural support or scaffold to facilitate function or enhanced function of the CD3 receptor complex. That is, the TCR may be any TCR and may not be utilized for its ability to target a particularly desired antigen. In such cases, and as an example, a TCR that targets a viral antigen may be employed for NK cells that will be used for cancers that are not necessarily related to that particular virus. In other cases, the TCR is selected for the ability to target a particular cancer antigen. Examples of antigens to which the TCR may be directed are provided elsewhere herein.

[0148] In FIG. 3, the following examples of constructs are noted:

[0149] TCR1: refers to TCRpp65 (the TCR against the HLA-A2 restricted CMVpp65) linked to the intracellular CD3zeta domain and full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon, and the construct may also be referred to as TCRpp65ZicdGDEFL that may comprise the following sequence:

MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVP NGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGI YGYTFGSGTRLTWEDLNKVFPP EVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALND SRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADRVK FSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQK DKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAGDVE ENPGPMILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDE KKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPA VYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSD FACANAFNNS I IPEDTFFPSPESSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKR RGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKD TYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAI ILLQGTLAQS IKGNH LVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQ NKSKPLQVYYRMCQNCIELNAATISGFLFAEIVS I FVLAVGVYFIAGQDGVRQSRASDKQTL LPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLV LATLLSQVSPFKIPIEELEDRVFVNCNTS ITWVEGTVGTLLSDITRLDLGKRILDPRGIYRC NGTDI YKDKESTVQVHYRMCQSCVELDPATVAGI IVTDVIATLLLALGVFCFAGHETGRLSG AADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWR VLGLCLLSVGVWGQDGNEEMGGITQTPYKVS ISGTTVILTCPQYPGSEILWQHNDKNIGGDE DDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVAT IVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKG QRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGPMRISKPHLRS IS IQCYLCLLLNSHFLT

EAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMK CFLLELQVISLESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSF VHIVQMFINTS* ( SEQ ID NO : 39 )

[0150] In TCRpp65ZicdGDEFL, the corresponding component sequences are as follows, although these particular sequences or others may be utilized in this and/or other constructs:

[0151] TCRb-extracellular domain'.

MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVP NGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGI YGYTFGSGTRLTWEDLNKVFPP EVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALND SRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD ( SEQ ID NO : 40 )

ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGAC CCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAA TGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCC AACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGC

CCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGAT ACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCC GAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGT GTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGG

AGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGAC AGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAA CCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACC GGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC ( SEQ ID

NO : 1 )

[0152] CD3 zeta intracellular domain (Z-ICD):

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNE LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAG DVEENPGP ( SEQ ID NO : 42 ) (where the P2A sequence is at the C- terminus )

AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTA TAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGG ACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAA CTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAG

GGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACG CCCTTCACATGCAGGCCCTGCCCCCTCGCgccaccaacttctccctgctgaagcaggccggc gacgtggaggagaaccccggcccc ( SEQ ID NO : 43 ) (where the lower case sequence is the P2A sequence )

[0153] TCRa-extracellular domain:

MILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGR ISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLR DSKSSDKSVCLFTDFDSQTNVSQSKDSDAYI TDKTVLDMRSMDFKSNSAVAWSNKSDFACAN AFNNS I I PEDTFFPSPESS ( SEQ ID NO : 44 )

ATGATCCTGAACGTGGAGCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAA CTTCACCTGCAGCTTCCCCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCG CCAAGAGCCCCGAGGCCCTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGG ATCAGCGCCACCCTGAACACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCC CGAGGACAGCGCCACCTACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCG GAACCAGCCTGACCGTGATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGG GACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGT GAGCCAGAGCAAGGACAGCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCA TGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAAC GCCTTCAACAACAGCATCATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGC ( SEQ ID NO : 45 )

[0154] CD3 gamma delta epsilon (CD3GDE):

MEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKMI GFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAAT I SGFLFAE IV S I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQT LNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKI PIEELEDRVFVNCNTS I TWV EGTVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAG I IVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWA RNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGI TQTPYKVS I S GTTVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGS KPEDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGGLLLLVYYWSKNRKAKAKPVTRG AGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNP GP ( SEQ ID NO : 4 6 ) (where the E2A sequence i s at the C-terminus )

ATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTT GGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGG TAG T T C T GAG T T GT GAT GCAGAAGCCAAAAATAT CACAT GGT T TAAAGAT GGGAAGAT GAT C GGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGG GAT G TAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T GTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTC AGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTC GAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATC GAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAGACC CTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAGCA CAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCAAGA TCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGGGTG GAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAATCCT GGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCACCG TGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCCGGC ATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGCCGG CCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACCAGG TGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGGGCC AGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGGCCC CATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGG GCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGC GGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAA CGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGA GCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGC AAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGA GATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGC TGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGC GCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCC CGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAA TCGGACCGcagtgtactaattatgctctcttgaaattggctggagatgttgagagcaatccc gggccc ( SEQ ID NO : 47 ) (where the lower case is the E2A sequence )

[0155] IL-15:

MRISKPHLRS IS IQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIED LIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDAS IHDTVENLI ILANNSLSS NGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS* ( SEQ ID NO : 48 )

ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCT GAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCG GACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGAC CTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTG CAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCG ACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGC AACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAA AGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 49 )

[0156] TCR2: refers to TCRpp65 linked to full length CD3zeta, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon; it lacks IL-15. Representative sequences are as follows:

CTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGACCCT GCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAATGG GACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCCAAC GGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGCCCC CAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGATACA CCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCCGAG GTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGTGTG CCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGGAGG TGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGACAGC CGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCA CTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACCGGG CCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGACGCCACCAACTTC AGCCTGCTGAAGCAGGCCGGCGACGTGGAGGAGAACCCCGGCCCCATGATCCTGAACGTGGA GCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAACTTCACCTGCAGCTTCC CCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCGCCAAGAGCCCCGAGGCC CTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGGATCAGCGCCACCCTGAA CACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCCCGAGGACAGCGCCACCT ACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCGGAACCAGCCTGACCGTG ATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGA CAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGAGCCAGAGCAAGGACA GCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAAC AGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCAT CATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGCGAGGGCAGAGGCAGCCTGCTGA CCTGCGGCGACGTGGAGGAGAACCCCGGCCCCATGAAGTGGAAGGCGCTTTTCACCGCGGCC ATCCTGCAGGCACAGTTGCCGATTACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACT CTGCTACCTGCTGGATGGAATCCTCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGA GAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTAT AACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGA CCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAAC TGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGG GGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGC CCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCG GCGACGTGGAGAGCAACCCCGGCCCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTG GCTATCATTCTTCTTCAAGGTACTTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGT G T AT GAG TAT C AAGAAGAT GGTTCGGTACTTCT GAG T T G T GAT G C AGAAG C C AAAAAT AT GA CAT GGT T TAAAGAT GGGAAGAT GAT CGGC T T CC TAAC T GAAGATAAAAAAAAAT GGAAT C T G GGAAGTAATGCCAAGGACCCTCGTGGGATGTATCAGTGTAAAGGATCACAGAACAAGTCAAA AC C AC T C C AAG T G T AT T AC AGAAT G T G T C AGAAC T G C AT T GAAC T AAAT G C AG C C AC CAT AT CTGGCTTTCTCTTTGCTGAAATCGTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATT GCTGGACAGGATGGAGTTCGCCAGTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGA CCAGCTCTACCAGCCCCTCAAGGATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACC AGTTGAGGAGGAATGTGAAGCAGACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTG GAGAGCAACCCCGGCCCCATGGAGCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCT GCTGAGCCAGGTGAGCCCCTTCAAGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGA ACTGCAACACCAGCATCACCTGGGTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACC AGACTGGACCTGGGCAAGAGAATCCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGA CATCTACAAGGACAAGGAGAGCACCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGG AGCTGGACCCCGCCACCGTGGCCGGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTG GCCCTGGGCGTGTTCTGCTTCGCCGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACAC CCAGGCCCTGCTGAGAAACGACCAGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGT ACAGCCACCTGGGCGGCAACTGGGCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGC GGCGACGTGGAGGAGAACCCCGGCCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCT GTGCCTGCTGAGCGTGGGCGTGTGGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCC AGACCCCCTACAAGGTGAGCATCAGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCC GGCAGCGAGATCCTGTGGCAGCACAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAA CATCGGCAGCGACGAGGACCACCTGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCT ACTACGTGTGCTACCCCAGAGGCAGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGA GCCAGAGTGTGCGAGAACTGCATGGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGT GGACATCTGCATCACCGGCGGCCTGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGG CCAAGGCCAAGCCCGTGACCAGAGGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAG GAGAGACCCCCCCCCGTGCCCAACCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCT

GTACAGCGGCCTGAACCAGAGAAGAATCGGACCG ( SEQ ID NO : 50 )

LEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGI TDQGEVPN GYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGI YGYTFGSGTRLTWEDLNKVFPPE VAVFEPSEAE I SHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDS RYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADATNF SLLKQAGDVEENPGPMILNVEQSPQSLHVQEGDSTNFTCS FPSSNFYALHWYRWETAKSPEA LFVMTLNGDEKKKGRI SATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTV I PNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYI TDKTVLDMRSMDFKSN SAVAWSNKSDFACANAFNNS I I PEDTFFPSPESSEGRGSLLTCGDVEENPGPMKWKALFTAA ILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLY NELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSE IGMKGERRR GKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLIL AI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNL GSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVS I FVLAVGVYFI AGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDV ESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITWVEGTVGTLLSDIT RLDLGKRILDPRGI YRCNGTDI YKDKESTVQVHYRMCQSCVELDPATVAGI IVTDVIATLLL

ALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTC GDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS ISGTTVILTCPQYP GSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLR ARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNK ERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGP ( SEQ ID NO : 51 )

[0157] TCR3: refers to TCRpp65 linked to the intracellular CD3z domain and IL-15, and it may also be referred to as TCRpp65Zicdl5, with a representative sequence as follows:

MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVP NGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGI YGYTFGSGTRLTWEDLNKVFPP EVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALND SRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADRVK FSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQK

DKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAGDVE ENPGPMILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDE KKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPA VYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSD FACANAFNNS I IPEDTFFPSPESSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKR

RGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKD T YDALHMQALPPRPGPQCTNYALLKLAGDVESNPGPMRI SKPHLRS I S I QCYLCLLLNSHFL TEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAM KCFLLELQVISLESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIKEFLQS FVHIVQMFINTS* ( SEQ ID NO : 52 )

[0158] In TCRpp65Zicdl5, the corresponding component sequences are as follows, although these particular sequences or others may be utilized in this and/or other constructs:

[0159] TCRb-extracellular domain:

ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGAC CCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAA TGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCC AACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGC CCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGAT

ACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCC GAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGT GTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGG AGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGAC AGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAA CCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACC GGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC ( SEQ ID NO : 41 )

[0160] CD3 zeta intracellular domain (Z-ICD):

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNE LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAG DVEENPGP ( SEQ ID NO : 42 ) (where P2A sequence is at the C- terminus )

GGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACG CCCTTCACATGCAGGCCCTGCCCCCTCGCgccaccaacttctccctgctgaagcaggccggc gacgtggaggagaaccccggcccc ( SEQ ID NO : 43 ) (where the lowercase sequence is P2A sequence )

[0161] TCRa-extracellular domain:

MILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGR ISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLR DSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSDFACAN AFNNS I IPEDTFFPSPESS ( SEQ ID NO : 44 )

ATGATCCTGAACGTGGAGCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAA CTTCACCTGCAGCTTCCCCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCG CCAAGAGCCCCGAGGCCCTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGG ATCAGCGCCACCCTGAACACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCC

CGAGGACAGCGCCACCTACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCG GAACCAGCCTGACCGTGATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGG GACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGT GAGCCAGAGCAAGGACAGCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCA

TGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAAC GCCTTCAACAACAGCATCATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGC ( SEQ ID NO : 45 )

[0162] CD3 zeta intracellular domain (Z-ICD)(m specific embodiments, two or more Z-

ICD sequences may be utilized):

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNE LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRPGPQCTNYALL KLAGDVESNPGP ( SEQ ID NO : 53 )

GGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACG CCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCC GGCGACGTGGAGAGCAACCCCGGCCCC ( SEQ ID NO : 54 )

[0163] IL-15:

ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCT GAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCG GACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGAC CTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTG CAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCG

ACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGC AACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAA AGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 49 )

[0164] TCR4: refers to TCRpp65 that also may be referred to as TCRpp65betaalpha, and a representative sequence is as follows:

RGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKD T YDALHMQALPPRPGPQCTNYALLKLAGDVESNPGPMRI SKPHLRS I S I QCYLCLLLNSHFL TEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAM KCFLLELQVISLESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIKEFLQS FVHIVQMFINTS* ( SEQ ID NO : 55 )

[0165] For TCRpp65betaalpha, the corresponding component sequences are as follows, although these particular sequences or others may be utilized in this and/or other constructs:

[0166] TCRb-extracellular domain:

ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGAC CCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAA TGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCC AACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGC CCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGAT ACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCC GAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGT GTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGG

AGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGAC AGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAA CCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACC GGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC ( SEQ ID NO : 41 )

[0167] CD 3 zeta intracellular domain (Z-ICD)'.

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNE LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAG DVEENPGP ( SEQ ID NO : 42 )

[0168] TCRa-extracellular domain'.

[0169] CD3 zeta intracellular domain (Z-ICD):

AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTA TAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGG ACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAA CTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAG GGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACG CCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCC GGCGACGTGGAGAGCAACCCCGGCCCC ( SEQ ID NO : 54 )

[0170] IL-15:

MRI SKPHLRS I S IQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVI SDLKKIED LIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVI SLESGDAS IHDTVENLI ILANNSLSS NGNVTESGCKECEELEEKNIKEFLQS FVHIVQMFINTS * ( SEQ ID NO : 48 )

[0171] An additional representative sequence for TCRpp65betaalpha is as follows:

ATGGACTCCTGGACCTTCTGCTGTGTGTCCCTTTGCATCCTGGTAGCAAAGCACACAGATGC TGGAGTTATCCAGTCACCCCGGCACGAGGTGACAGAGATGGGACAAGAAGTGACTCTGAGAT GTAAACCAATTTCAGGACACGACTACCTTTTCTGGTACAGACAGACCATGATGCGGGGACTG GAGTTGCTCATTTACTTTAACAACAACGTTCCGATAGATGATTCAGGGATGCCCGAGGATCG ATTCTCAGCTAAGATGCCTAATGCATCATTCTCCACTCTGAAGATCCAGCCCTCAGAACCCA GGGACTCAGCTGTGTACTTCTGTGCCAGCAGTTCGGCAAACTATGGCTACACCTTCGGTTCG GGGACCAGGTTAACCGTTGTAGAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGTGTT TGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAG GCTTCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGG GTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCT GAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTC AAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCCGTC ACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTTACCTCGGTGTCCTA CCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAGGCCACCCTGT ATGCTGTGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAAGAGAAAGGATTTCGAGGGC AGGGGAAGTCTTCTAACATGCGGGGACGTGGAGGAAAATCCCGGGCCCATGCTCCTTGAACA TTTATTAATAATCTTGTGGATGCAGCTGACATGGGTCAGTGGTCAACAGCTGAATCAGAGTC C T CAAT C TAT G T T TAT C CAGGAAGGAGAAGAT G T C T C CAT GAAC T GCAC T T C T T CAAGCAT A TTTAACACCTGGCTATGGTACAAGCAGGACCCTGGGGAAGGTCCTGTCCTCTTGATAGCCTT ATATAAGGCTGGTGAATTGACCTCAAATGGAAGACTGACTGCTCAGTTTGGTATAACCAGAA AGGACAGCTTCCTGAATATCTCAGCATCCATACCCAGTGATGTAGGCATCTACTTCTGTGCT GGACCCATGAAAACCTCCTACGACAAGGTGATATTTGGGCCAGGGACAAGCTTATCAGTCAT T C C AAAT AT C C AGAAC C C T GAG C C T G C C G T G T AC C AG C T GAGAGAC T C T AAAT C C AG T GAG A AGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCT GAT G T G TAT AT CACAGACAAAAC T G T GC T AGACAT GAGG T C T AT GGAC T T CAAGAGCAACAG TGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTA TTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAA AGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCT CCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGA ( SEQ ID NO : 56 )

MDS WT FCCVS LG I LVAKHTDAGVI QS PRHEVTEMGQEVTLRCKP I S GHDYL FWYRQTMMRGL ELLIYFNNNVPIDDSGMPEDRFSAKMPNAS FSTLKIQPSEPRDSAVYFCASSSANYGYTFGS GTRLTWEDLNKVFPPEVAVFEPSEAE I SHTQKATLVCLATGFFPDHVELSWWVNGKEVHSG VSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSVSYQQGVLSAT ILYE ILLGKATLYAVLVSALVLMAMVKRKDFEG RGSLLTCGDVEENPGPMLLEHLLI ILWMQLTWVSGQQLNQSPQSMFIQEGEDVSMNCTSSS I FNTWLWYKQDPGEGPVLLIALYKAGELTSNGRLTAQFGI TRKDS FLNI SAS I PSDVGIYFCA GPMKTSYDKVI FGPGTSLSVI PNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDS DVYI TDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNS I I PEDTFFPSPESSCDVKLVEK S FETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS * ( SEQ ID NO : 57 )

[0172] Zl: refers to full length CD3zeta, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to IL15 (see FIGS. 2A and 2B), and it may also be referred to as CD3ZFLGDEFL15, and representative sequences may be as follows:

MLEMKWKALFTAAILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSA DAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE AYSE IGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPG PMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKM IGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAAT I SGFLFAE I VS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQ TLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKI PIEELEDRVFVNCNTS I TW VEGTVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVA GI IVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNW ARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGI TQTPYKVS I SGTTVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRG SKPEDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGGLLLLVYYWSKNRKAKAKPVTR GAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESN PGPMRI SKPHLRS I S IQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVI SDLKK IEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVI SLESGDAS IHDTVENLI ILANNS LSSNGNVTESGCKECEELEEKNIKEFLQS FVHIVQMFINTS ( SEQ ID NO : 58 )

ATGCTCGAGATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGAT TACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCC TCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCA GACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAG AGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGC AGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAG GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTA CCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCC CTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGC CCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTAC TTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTT CGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATG ATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCG TGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAA TGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATC GTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCA GTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGG ATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAG ACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGA GCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCA AGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGG GTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAAT CCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCA CCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCC

GGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGC CGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACC AGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGG GCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGG CCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGT GGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATC AGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCA CAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACC

TGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGC AGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCAT GGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCC TGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGA GGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAA CCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAA GAATCGGACCGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAAT CCCGGGCCCATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTG

CCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCT TCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAG ATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCA CCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGG AAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGC CTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAA GAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 59 )

[0173] Z2: refers to full length CD3zeta, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to membrane bound IL21 (with CD8 transmembrane domain for the membrane bound IL21), and it may also be referred to as CD3ZGDEFLSP821CD28, and a representative sequence is as follows:

MLEMKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSA DAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPG PMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKM IGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEI VS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQ TLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITW VEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVA

GI IVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNW ARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS I SGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRG SKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTR GAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESN PGPMR I CL T S DRLAPAAGLAAPRRQAVHKS S S QGQDRHM I RMRQL I D I VDQLKNYVNDLVPE FLPAPEDVETNCEWSAFSCFQKAQLKSANTGNNERI INVS IKKLKRKPPSTNAGRRQKHRLT CPSCDSYEKKPPKEFLERFKSLLQKMIHQHLSSRTHGSEDSTTTPAPRPPTPAPT IASQPLS LRPEACRPAAGGAVHTRGLDFACDFWVLVWGGVLACYSLLVTVAFI I FWV* ( SEQ ID

NO : 60 )

[0174] For CD3ZGDEFLSP821CD28, the corresponding component sequences are as follows, although these particular sequences or others may be utilized in this and/or other constructs:

[0175] CD3

MLEMKWKALFTAAILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSA DAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE AYSE IGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPG PMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKM IGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAAT I SGFLFAE I VS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQ TLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKI PIEELEDRVFVNCNTS I TW VEGTVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVA GI IVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNW ARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGI TQTPYKVS I SGTTVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRG

SKPEDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGGLLLLVYYWSKNRKAKAKPVTR GAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESN PGP ( SEQ ID NO : 61 )

ATGCTCGAGATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGAT TACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCC TCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCA GACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAG AGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGC AGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAG GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTA CCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCC CTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGC CCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTAC TTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTT

CGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATG ATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCG TGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC C AC T C C AAG T G T AT T AC AGAA TGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATC GTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCA GTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGG ATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAG ACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGA GCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCA AGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGG GTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAAT

CCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCA CCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCC GGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGC CGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACC AGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGG GCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGG CCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGT GGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATC AGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCA CAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACC TGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGC AGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCAT GGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCC TGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGA

GGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAA CCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAA GAATCGGACCGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAAT CCCGGGCCC ( SEQ ID NO : 62 )

[0176] SP CD8

MRICLTSDRLAPAAGLAAPRRQAV ( SEQ ID NO : 63 ) atgcgcatttgcctgaccagcgatcgcctggcgccggcggcgggcctggcggcgccgcgccg ccaggcggtg ( SEQ ID NO : 64 )

[0177] IL-21:

HKSSSQGQDRHMIRMRQLIDIVDQLKNYVNDLVPEFLPAPEDVETNCEWSAFSCFQKAQLKS ANTGNNERI INVS IKKLKRKPPSTNAGRRQKHRLTCPSCDSYEKKPPKEFLERFKSLLQKMI HQHLSSRTHGSEDS ( SEQ ID NO : 65 )

CATAAATCTTCCTCTCAAGGTCAGGACCGCCATATGATTCGAATGCGGCAGCTGATTGACAT AGTCGATCAACTGAAGAACTATGTGAATGATCTTGTGCCCGAGTTTTTGCCAGCCCCTGAAG ACGTAGAAACTAATTGTGAGTGGAGTGCCTTTTCCTGCTTTCAAAAGGCACAGCTGAAATCC GCCAACACGGGCAATAACGAACGGATAATTAACGTATCCATTAAGAAGCTGAAGCGGAAGCC GCCCTCAACCAATGCGGGACGGCGGCAAAAGCATCGCTTGACCTGTCCGTCATGCGACAGCT AC GAGAAAAAG C C C C C GAAG GAG T T C T T G GAAC G C T T C AAGAG T C T C C T T C AGAAAAT GAT T

CACCAGCACCTGTCCTCACGGACGCACGGAAGCGAGGACAGT ( SEQ ID NO : 66 )

[0178] CD8 hinge-.

TTTPAPRPPTPAPT IASQPLSLRPEACRPAAGGAVHTRGLDFACD ( SEQ ID NO : 67 )

ACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACCATCGCGTCGCAGCCCCTGTC CCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACT TCGCCTGTGAT ( SEQ ID NO : 68 )

[0179] CD28 Transmembrane domain'.

FWVLVWGGVLACYSLLVTVAFI I FWV* ( SEQ ID NO : 69 )

TTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGC CTTTATTATTTTCTGGGTG ( SEQ ID NO : 70 ) [0180] Z3: refers to full length CD3zeta, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to membrane bound IL21 (with CD28 transmembrane domain for the membrane bound IL21), and it may also be referred to as CD3ZGDEFL8SP21CD8 with a representative sequence as follows:

MLEMKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSA DAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPG PMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKM IGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEI VS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQ

TLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITW VEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVA GI IVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNW ARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS I SGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRG

SKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTR GAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESN PGPMR I CL T S DRLAPAAGLAAPRRQAVHKS S S QGQDRHM I RMRQL I D I VDQLKNYVNDLVPE FLPAPEDVETNCEWSAFSCFQKAQLKSANTGNNERI INVS IKKLKRKPPSTNAGRRQKHRLT CPSCDSYEKKPPKEFLERFKSLLQKMIHQHLSSRTHGSEDSTTTPAPRPPTPAPTIASQPLS

LRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVIT ( SEQ ID NO : 71 ) .

[0181] For CD3ZGDEFL8SP21CD8, the corresponding component sequences are as follows, although these particular sequences or others may be utilized in this and/or other constructs:

[0182] CD3:

SKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTR GAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESN PGP ( SEQ ID NO : 61 )

ATGCTCGAGATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGAT TACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCC TCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCA GACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAG AGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGC

AGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAG GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTA CCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCC CTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGC CCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTAC TTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTT CGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATG ATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCG TGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAA TGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATC GTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCA GTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGG ATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAG ACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGA GCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCA AGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGG GTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAAT CCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCA CCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCC GGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGC CGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACC AGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGG GCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGG CCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGT GGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATC AGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCA CAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACC TGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGC AGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCAT GGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCC TGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGA GGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAA CCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAA GAATCGGACCGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAAT CCCGGGCCC ( SEQ ID NO : 62 )

[0183] SP CD8:

[0184] IL-21 :

HKSSSQGQDRHMIRMRQLIDIVDQLKNYVNDLVPEFLPAPEDVETNCEWSAFSCFQKAQLKS ANTGNNERI INVS IKKLKRKPPSTNAGRRQKHRLTCPSCDSYEKKPPKEFLERFKSLLQKMI HQHLSSRTHGSEDS ( SEQ ID NO : 65 ) cataaatcttcctctcaaggtcaggaccgccatatgattcgaatgcggcagctgattgacat agtcgatcaactgaagaactatgtgaatgatcttgtgcccgagtttttgccagcccctgaag acgtagaaactaattgtgagtggagtgccttttcctgctttcaaaaggcacagctgaaatcc gccaacacgggcaataacgaacggataattaacgtatccattaagaagctgaagcggaagcc gccctcaaccaatgcgggacggcggcaaaagcatcgcttgacctgtccgtcatgcgacagct acgagaaaaagcccccgaaggagttcttggaacgcttcaagagtctccttcagaaaatgatt caccagcacctgtcctcacggacgcacggaagcgaggacagt ( SEQ ID NO : 66 )

[0185] CD8 hinge:

TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD ( SEQ ID NO : 67 )

[0186] CD8 Transmembrane Domain:

IYIWAPLAGTCGVLLLSLVIT* ( SEQ ID NO : 72 )

ATCTACATCTGGGCGCCCTTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTGGTTATCAC C ( SEQ ID NO : 73 )

[0187] In certain embodiments, provided herein are CD3 constructs comprising a fusion with an intracellular co-stimulatory domain derived from CD 16, NKG2D, DAP 10, DAP 12, 2B4, 4- IBB, CD2, CD28, DNAM, or any combination thereof. In certain embodiments, an intracellular co-stimulatory domain is fused to CD36, CD3s, CD3y, and/or CD3^. In certain embodiments, such a CD3 fusion construct comprises a CD3^ fused to a DAP 10 intracellular co-stimulatory domain. In certain embodiments, such a CD3 fusion construct comprises a CD3^ fused to a CD28 intracellular co-stimulatory domain. In certain embodiments, such a CD3 fusion construct comprises a CD3^ fused to a DAP 10 intracellular co-stimulatory domain and a CD28 intracellular co-stimulatory domain. In certain embodiments, a CD3^ fused to a DAP 10 intracellular co-stimulatory domain is represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 106. In certain embodiments, a CD3^ fused to a CD28 intracellular co-stimulatory domain is represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 107. In certain embodiments, a CD3^ fused to a DAP10 intracellular co-stimulatory domain and a CD28 intracellular co-stimulatory domain is represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108. In certain embodiments, a CD3^ fused to a DAP10 intracellular co-stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109. In certain embodiments, a CD3^ fused to a CD28 intracellular co-stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 110. In certain embodiments, a CD3^ fused to a DAP10 intracellular co-stimulatory domain and a CD28 intracellular costimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 111. In certain embodiments, a CD3^ fused to an intracellular domain may not comprise a C terminal 2A domain. In certain embodiments, a CD3^ fused to an intracellular domain may not comprise an N terminal signal peptide domain.

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGCTTTGCGCACGCCCACGCCGCAGCCCCGCCCAA GAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGC AGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAA GAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCG CAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGA GGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTT ACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCC CCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGG CCCC ( SEQ ID NO : 106 )

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGAC TACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCAAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCG CGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTAC GATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAA GAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTG AGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTC AGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTG CACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC ( SEQ ID NO : 107 )

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGAC TACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCACTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAG AAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGCA GACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAG AGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGC AGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAG GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTA CCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCC CTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGC CCC ( SEQ ID NO : 108 )

MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLLCARPRRSPAQ EDGKVYINMPGRGRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKP QRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALP PRQCTNYALLKLAGDVESNPGP ( SEQ ID NO : 109 )

MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSD YMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGL STATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGP ( SEQ ID NO : 110 )

MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSD

YMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMPGRGRVKFSRSA

DAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE

AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPG P ( SEQ ID NO : 111 )

[0188] In certain embodiments, a DAP 10 intracellular co-stimulatory domain is represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 112. In certain embodiments, a CD28 intracellular co-stimulatory domain is represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 113. In certain embodiments, a DAP10 intracellular co-stimulatory domain and CD28 intracellular co-stimulatory domain is represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 114. In certain embodiments, a DAP10 intracellular co-stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 115. In certain embodiments, a CD28 intracellular co-stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 116. In certain embodiments, a DAP10 intracellular co-stimulatory domain and CD28 intracellular co-stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 117.

CTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCC AGGCAGGGGC ( SEQ ID NO : 112 )

AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGG GCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCA ( SEQ ID NO : 113 )

AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGG GCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCAC TTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCCA GGCAGGGGC ( SEQ ID NO : 114 )

LCARPRRSPAQEDGKVYINMPGRG ( SEQ ID NO : 115 )

RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS ( SEQ ID NO : 116 )

RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMP GRG ( SEQ ID NO : 117 )

[0189] UTNK15-D AP 10 : refers to full length CD3zeta comprising a fusion with an intracellular co-stimulatory domain derived from DAP10, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to IL 15, it may be represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 118. In certain embodiments, a UTNK15-DAP10 amino acid sequence may be represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 119.

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGCTTTGCGCACGCCCACGCCGCAGCCCCGCCCAA GAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGC AGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAA GAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCG CAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGA GGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTT ACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCC CCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGG CCCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTA CTTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGT TCGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGAT GATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTC GTGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT TAG AGA ATGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAAT CGTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCC AGTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAG GATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCA GACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGG AGCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTC AAGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTG GGTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAA TCCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGC ACCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGC CGGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCG CCGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGAC CAGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTG GGCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCG GCCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTG TGGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCAT CAGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGC ACAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCAC CTGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGG CAGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCA TGGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGC CTGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAG AGGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCA ACCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGA AGAATCGGACCGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAA TCCCGGGCCCATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGT GCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGC TTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAA GATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGC ACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTG GAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAG CCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGA AGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 118 )

MKWKALFTAAILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLLCARPRRSPAQ EDGKVYINMPGRGRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKP QRRKNPQEGLYNELQKDKMAEAYSE IGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALP PRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDG SVLLTCDAEAKNI TWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYR MCQNCIELNAAT I SGFLFAE IVS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLK DREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPF KI PIEELEDRVFVNCNTS I TWVEGTVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKES T VQVH YRMC Q S C VE L D PAT VAG I IVTDVIATLL LAL GVFC FAGHE T GRL S GAAD T QAL LRND QVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGV WGQDGNEEMGGI TQTPYKVS I SGTTVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDH LSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGG LLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQR RIGPQCTNYALLKLAGDVESNPGPMRI SKPHLRS I S IQCYLCLLLNSHFLTEAGIHVFILGC FSAGLPKTEANWVNVI SDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVI SL ESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIKEFLQS FVHIVQMFINTS ( SEQ ID NO : 119 )

[0190] UTNK15-28: refers to full length CD3zeta comprising a fusion with an intracellular co-stimulatory domain derived from CD28, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to IL15, it may be represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 120. In certain embodiments, a UTNK 15-28 amino acid sequence may be represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 121.

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGAC TACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCAAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCG CGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTAC GATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAA GAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTG AGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTC AGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTG CACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAAC AGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTTGGCCCAG TCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGGTACTTCT GACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATGATCGGCTTCC TAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T G T C AGAA CTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTCAGCATTT TCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTCGAGAGCT TCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATCGAGAAGA

TGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAGACCCTGAACT TCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAGCACAGCACC TTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCAAGATCCCCAT CGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGGGTGGAGGGCA CCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAATCCTGGACCCC AGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCACCGTGCAGGT GCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCCGGCATCATCG TGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGCCGGCCACGAG ACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACCAGGTGTACCA GCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGGGCCAGAAACA AGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGGCCCCATGCAG AGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGGGCCAGGA CGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGCGGCACCA CCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAACGACAAG AACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGAGCCTGAA GGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGCAAGCCCG AGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGAGATGGAC GTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGCTGCTGCT GGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGCGCCGGCG CCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCCCGACTAC GAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAATCGGACC GCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCA TGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTG AACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGG ACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACC TGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGC AAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGA CGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCA ACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAA GAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 120 )

MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSD YMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGL STATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAI ILLQGTLAQ S IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMY QCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVS I FVLAVGVYFIAGQDGVRQSRA SDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHST FLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITWVEGTVGTLLSDITRLDLGKRILDP RGI YRCNGTDI YKDKESTVQVHYRMCQSCVELDPATVAGI IVTDVIATLLLALGVFCFAGHE TGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQ SGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS ISGTTVILTCPQYPGSEILWQHNDK NIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD VMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDY EPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGPMRISKPHLRS IS IQCYLCLLL NSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSC KVTAMKCFLLELQVISLESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIK EFLQSFVHIVQMFINTS ( SEQ ID NO : 121 )

[0191] UTNK15-28-DAP10: refers to full length CD3zeta comprising a fusion with an intracellular co- stimulatory domain derived from DAP 10 and an intracellular co-stimulatory domain derived from CD28, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to IL15, it may be represented by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122. In certain embodiments, a UTNK15-28-DAP10 amino acid sequence may be represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123. ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGAC TACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCACTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAG AAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGCA GACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAG AGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGC AGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAG GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTA CCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCC CTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGC CCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTAC TTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTT CGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATG ATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCG TGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAA TGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATC GTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCA GTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGG ATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAG ACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGA GCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCA AGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGG GTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAAT CCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCA CCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCC GGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGC CGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACC AGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGG GCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGG CCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGT GGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATC AGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCA CAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACC TGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGC AGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCAT GGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCC TGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGA GGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAA CCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAA GAATCGGACCGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAAT CCCGGGCCCATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTG CCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCT TCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAG ATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCA CCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGG AAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGC CTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAA GAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 122 ) MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSD YMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMPGRGRVKFSRSA DAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPG PMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKM IGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEI VS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQ TLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITW VEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVA GI IVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNW ARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS I SGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRG SKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTR GAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESN PGPMRISKPHLRS IS IQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKK IEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDAS IHDTVENLI ILANNS

LSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS ( SEQ ID NO : 123 )

[0192] As depicted in FIG. 3 and described above, the term “linked” refers to being present on the same polynucleotide vector and does not necessarily mean that the two polypeptides are expressed as one polypeptide. For example, a cytokine produced from a vector of the disclosure may ultimately be produced as a separate molecule from any one or more TCR/CD3 receptor complex components. Whereas, the term “fused” or “fusion” refers to two polypeptides that comprise a peptide bond conjoining the two molecules, i.e. that the two polypeptides are covalently bound by an amide bond and are not separated by a splitting element, such as a 2A element.

[0193] One specific example of a TCR that may be utilized in the cells is NY-ESO TCR, and specific examples of sequences include at least the following:

[0194] TCRa:

XQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRL NASLDKSSGRSTLYIAASQPGDSATYLCAVRPLYGGSYIPTFGRGTSLIVHPYIQNPDPAVY QLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFA CANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLM TLRLWSS ( SEQ ID NO : 25 )

[0195] TCRP:

GVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGY NVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVLEDLKNVFPPKVAV EE P S E AE I S H T QKAT L VC LAT G F Y P DHVE L S WWVNGKE VH SGVSTDPQPLKEQ PALND S R YC LSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSES YQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 26 ) [0196] In certain embodiments, a TCR may comprise a TCR alpha chain variable region encoded by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 85. aaacaggaggtgacacagattcctgcagctctgagtgtcccagaaggagaaaacttggttct caactgcagtttcactgatagcgctatttacaacctccagtggtttaggcaggaccctggga aaggtctcacatctctgttgcttattcagtcaagtcagagagagcaaacaagtggaagactt aatgcctcgctggataaatcatcaggacgtagtactttatacattgcagcttctcagcctgg tgactcagccacctacct ctgtgctgtgaggcccctttatggaggaagctacatacctacat ttggaagaggaaccagccttattgttcatccgtat ( SEQ ID NO : 85 )

[0197] In certain embodiments, a TCR may comprise a TCR alpha chain constant region encoded by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 86. atccagaaccctgaccctgccgtgtaccagctgagagactctaaatccagtgacaagtctgt ctgcctattcaccgattttgattctcaaacaaatgtgtcacaaagtaaggattctgatgtgt atatcacagacaaaactgtgctagacatgaggtctatggacttcaagagcaacagtgctgtg gcctggagcaacaaatctgactttgcatgtgcaaacgccttcaacaacagcattattccaga agacaccttcttccccagcccagaaagttcctgtgatgtcaagctggtcgagaaaagctttg aaacagatacgaacctaaactttcaaaacctgtcagtgattgggttccgaatcctcctcctg aaagtggccgggtttaatctgctcatgacgctgcggctgtggtccagc ( SEQ ID NO : 86 )

[0198] In certain embodiments, a TCR may comprise a TCR alpha chain encoded by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87. atggagaccctcttgggcctgcttatcctttggctgcagctgcaatgggtgagcagcaaaca ggaggtgacacagattcctgcagctctgagtgtcccagaaggagaaaacttggttctcaact gcagtttcactgatagcgctatttacaacctccagtggtttaggcaggaccctgggaaaggt ctcacatctctgttgcttattcagtcaagtcagagagagcaaacaagtggaagacttaatgc ctcgctggataaatcatcaggacgtagtactttatacattgcagcttctcagcctggtgact cagccacctacct ctgtgctgtgaggcccctttatggaggaagctacatacctacatttgg a agaggaaccagccttattgttcatccgtatatccagaaccctgaccctgccgtgtaccagct gagagactctaaatccagtgacaagtctgtctgcctattcaccgattttgattctcaaacaa atgtgtcacaaagtaaggattctgatgtgtatatcacagacaaaactgtgctagacatgagg tctatggacttcaagagcaacagtgctgtggcctggagcaacaaatctgactttgcatgtgc aaacgccttcaacaacagcattattccagaagacaccttcttccccagcccagaaagttcct gtgatgtcaagctggtcgagaaaagctttgaaacagatacgaacctaaactttcaaaacctg tcagtgattgggttccgaatcctcctcctgaaagtggccgggtttaatctgctcatgacgct gcggctgtggtccagc ( SEQ ID NO : 87 )

[0199] In certain embodiments, a TCR may comprise a TCR alpha chain variable region amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ

ID NO: 88.

XQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRL NASLDKSSGRSTLYIAASQPGDSATYLCAVRPLYGGSYIPTFGRGTSLIVHPY ( SEQ ID NO : 88 )

[0200] In certain embodiments, a TCR may comprise a TCR alpha chain constant region amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 89.

IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAV AWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLL KVAGFNLLMTLRLWSS ( SEQ ID NO : 89 )

[0201] In certain embodiments, a TCR may comprise an alpha chain CDR1 amino acid sequence that is at least, or exactly, 80% or 100% identical to SEQ ID NO: 90.

DSAIYN ( SEQ ID NO : 90 )

[0202] In certain embodiments, a TCR may comprise an alpha chain CDR2 amino acid sequence that is at least, or exactly, 80% or 100% identical to SEQ ID NO: 91.

IQSSQRE ( SEQ ID NO : 91 )

[0203] In certain embodiments, a TCR may comprise an alpha chain CDR3 amino acid sequence that is at least, or exactly, 80% or 100% identical to SEQ ID NO: 92.

CAVRPLYGGSYIPTF ( SEQ ID NO : 92 )

[0204] In certain embodiments, a TCR may comprise a TCR beta chain variable encoded by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 93. ggtgtcactcagaccccaaaattccaggtcctgaagacaggacagagcatgacactgcagtg tgcccaggatatgaaccatgaatacatgtcctggtatcgacaagacccaggcatggggctga ggctgattcattactcagttggtgctggtatcactgaccaaggagaagtccccaatggctac aatgtctccagatcaaccacagaggatttcccgctcaggctgctgtcggctgctccctccca gacatctgtgtactt ctgtgccagcagttacgtcgggaacaccggggagctgttttttgg a g aaggctctaggctgaccgtactggag ( SEQ ID NO : 93 )

[0205] In certain embodiments, a TCR may comprise a TCR beta chain constant region encoded by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94. Gacctgaaaaacgtgttcccacccaaggtcgctgtgtttgagccatcagaagcagagatctc ccacacccaaaaggccacactggtatgcctggccacaggcttctaccccgaccacgtggagc tgagctggtgggtgaatgggaaggaggtgcacagtggggtcagcacagacccgcagcccctc aaggagcagcccgccctcaatgactccagatactgcctgagcagccgcctgagggtctcggc caccttctggcagaacccccgcaaccacttccgctgtcaagtccagttctacgggctctcgg agaatgacgagtggacccaggatagggccaaacccgtcacccagatcgtcagcgccgaggcc tggggtagagcagactgtggcttcacctccgagtcttaccagcaaggggtcctgtctgccac catcctctatgagatcttgctagggaaggccaccttgtatgccgtgctggtcagtgccctcg tgctgatggccatggtcaagagaaaggattccagaggc ( SEQ ID NO : 94 )

[0206] In certain embodiments, a TCR may comprise a TCR beta chain encoded by a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ

ID NO: 95.

At gagcat cggcctcctgt get gtgcagccttgtctctcctgtgggcaggtccagtgaatgc tggtgtcactcagaccccaaaattccaggtcctgaagacaggacagagcatgacactgcagt gtgcccaggatatgaaccatgaatacatgtcctggtatcgacaagacccaggcatggggctg aggctgattcattactcagttggtgctggtatcactgaccaaggagaagtccccaatggcta caatgtctccagatcaaccacagaggatttcccgctcaggctgctgtcggctgctccctccc agacatctgtgtactt ctgtgccagcagttacgtcgggaacaccggggagctgttttttgg a gaaggctctaggctgaccgtactggaggacctgaaaaacgtgttcccacccAaggtcgctgt gtttgagccatcagaagcagagatctcccacacccaaaaggccacactggtatgcctggcca caggcttctaccccgaccacgtggagctgagctggtgggtgaatgggaaggaggtgcacagt ggggtcagcacagacccgcagcccctcaaggagcagcccgccctcaatgactccagatactg cctgagcagccgcctgagggtctcggccaccttctggcagaacccccgcaaccacttccgct gtcaagtccagttctacgggctctcggagaatgacgagtggacccaggatagggccaaaccc gtcacccagatcgtcagcgccgaggcctggggtagagcagactgtggcttcacctccgagtc ttaccagcaaggggtcctgtctgccaccat octet at gagatett get agggaaggccacct tgtatgccgtgctggtcagtgccctcgtgctgatggccatggtcaagagaaaggattccaga ggc ( SEQ ID NO : 95 )

[0207] In certain embodiments, a TCR may comprise a TCR beta chain variable region amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ

ID NO: 96.

GVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGY NVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVLE ( SEQ ID NO : 96 )

[0208] In certain embodiments, a TCR may comprise a TCR beta chain constant region amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ

ID NO: 97.

DLKNVFPPKVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPL KEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEA WGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 97 )

[0209] In certain embodiments, a TCR may comprise a beta chain CDR1 amino acid sequence that is at least, or exactly, 80% or 100% identical to SEQ ID NO: 98.

MNHEY ( SEQ ID NO : 98 )

[0210] In certain embodiments, a TCR may comprise a beta chain CDR2 amino acid sequence that is at least, or exactly, 80% or 100% identical to SEQ ID NO: 99.

SVGAGI ( SEQ ID NO : 99 )

[0211] In certain embodiments, a TCR may comprise a beta chain CDR3 amino acid sequence that is at least, or exactly, 80% or 100% identical to SEQ ID NO: 100.

CASSYVGNTGELFF ( SEQ ID NO : 100 )

[0212] In certain embodiments, a TCR (e.g., a TCR alpha, beta, delta, and/or gamma) chain may comprise a signal peptide. In certain embodiments, a signal peptide is encoded by a nucleic acid that is at least, or exactly 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 or SEQ ID NO: 102. In certain embodiments, a signal peptide is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 103 or SEQ ID NO: 104. atggagaccctcttgggcctgcttatcctttggctgcagctgcaatgggtgagcagc ( SEQ ID NO : 101 ) at gagcat cggcctcctgt get gtgcagccttgtctctcctgtgggcaggtccagtgaatgc t ( SEQ ID NO : 102 )

METLLGLLILWLQLQWVSS ( SEQ ID NO : 103 )

MS IGLLCCAALSLLWAGPVNA ( SEQ ID NO : 104 )

[0213] In certain embodiments, a TCR recognizes a peptide corresponding to amino acid residues 157-165 of the human cancer testis Ag NY-ESO-1 in the context of the HLA-A*02 class I allele. In certain embodiments, a TCR may target an epitope characterized by the amino acid sequence according to SEQ ID NO: 105.

SLLMWITQC ( SEQ ID NO : 105 )

[0214] One specific example of a TCR that may be utilized in the cells is TCRpp65alpha, and specific examples of sequences include at least the following (underlining refers to signal peptide sequence): ATGGACTCCTGGACCTTCTGCTGTGTGTCCCTTTGCATCCTGGTAGCAAAGCACACAGATGC TGGACAACAG C T GAAT C AGAG T C C T C AAT CTATGTTTATC GAG GAAG GAGAAGAT G T C T C C A TGAACTGCACTTCTTCAAGCATATTTAACACCTGGCTATGGTACAAGCAGGACCCTGGGGAA GGTCCTGTCCTCTTGATAGCCTTATATAAGGCTGGTGAATTGACCTCAAATGGAAGACTGAC TGCTCAGTTTGGTATAACCAGAAAGGACAGCTTCCTGAATATCTCAGCATCCATACCCAGTG ATGTAGGCATCTACTTCTGTGCTGGACCCATGAAAACCTCCTACGACAAGGTGATATTTGGG CCAGGGACAAGCTTATCAGTCATTCCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCT GAGAGAC T C T AAAT C C AG T GAG AAG T C T G T C T G C C T AT T C AC C GAT T T T GAT T C T C AAAC AA AT G T G T C AC AAAG T AAG GATTCTGATGTGTATAT C AC AGAC AAAAC T G T G C T AGAC AT GAG G TCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGC AAAC G C C T T C AAC AAC AG CAT TAT T C C AGAAGAC AC CTTCTTCCC C AG C C C AGAAAG T T C C T G T GAT G T C AAG C T G G T C GAGAAAAG C T T T GAAACAGAT AC GAAC C T AAAC T T T C AAAAC C T G TCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCT GCGGCTGTGGTCCAGC ( SEQ ID NO : 27 )

MDSWTFCCVSLCILVAKHTDAGQQLNQS PQSMFI QEGEDVSMNCTS S S I FNTWLWYKQDPGE GPVLLIALYKAGELTSNGRLTAQFGI TRKDS FLNI SAS I PSDVGIYFCAGPMKTSYDKVI FG PGTSLSVI PNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYI TDKTVLDMR SMDFKSNSAVAWSNKSDFACANAFNNS I I PEDTFFPSPESSCDVKLVEKS FETDTNLNFQNL SVIGFRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 28 )

[0215] One specific example of a TCR that may be utilized in the cells is TCRpp65beta, and specific examples of sequences include at least the following (underlining refers to signal peptide sequence):

ATGGACTCCTGGACCTTCTGCTGTGTGTCCCTTTGCATCCTGGTAGCAAAGCACACAGATGC TGGAGTTATCCAGTCACCCCGGCACGAGGTGACAGAGATGGGACAAGAAGTGACTCTGAGAT GTAAACCAATTTCAGGACACGACTACCTTTTCTGGTACAGACAGACCATGATGCGGGGACTG GAGTTGCTCATTTACTTTAACAACAACGTTCCGATAGATGATTCAGGGATGCCCGAGGATCG ATTCTCAGCTAAGATGCCTAATGCATCATTCTCCACTCTGAAGATCCAGCCCTCAGAACCCA GGGACTCAGCTGTGTACTTCTGTGCCAGCAGTTCGGCAAACTATGGCTACACCTTCGGTTCG GGGACCAGGTTAACCGTTGTAGAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGTGTT TGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAG GCTTCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGG GTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCT GAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTC AAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCCGTC ACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTTACCTCGGTGTCCTA CCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAGGCCACCCTGT ATGCTGTGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAAGAGAAAGGATTTC ( SEQ ID NO : 29 )

MPS WT FCCVS LC I LVAKHTDAGVI QS PRHEVTEMGQEVTLRCKP I S GHDYL FWYRQTMMRGL ELLIYFNNNVPIDDSGMPEDRFSAKMPNAS FSTLKIQPSEPRDSAVYFCASSSANYGYTFGS GTRLTWEDLNKVFPPEVAVFEPSEAE I SHTQKATLVCLATGFFPDHVELSWWVNGKEVHSG VSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSVSYQQGVLSAT ILYE ILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 30 )

[0216] TCRpp65ZFLGDEFL15 [0217] In certain embodiments, one may utilize a construct in which TCRpp65 is linked to full length CD3zeta, full length CD3 gamma, full length CD3 delta, full length CD3 epsilon, and also linked to IL- 15 (and may be referred to as TCRpp65ZFLGDEFL15). One representative sequence for such a construct is as follows:

MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVP NGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGI YGYTFGSGTRLTWEDLNKVFPP EVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALND SRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADATN FSLLKQAGDVEENPGPMILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPE ALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLT VIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKS NSAVAWSNKSDFACANAFNNS I IPEDTFFPSPESSEGRGSLLTCGDVEENPGPMKWKALFTA AILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQL YNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERR RGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLI LAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWN LGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVS I FVLAVGVYF IAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGD VESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITWVEGTVGTLLSDI TRLDLGKRILDPRGI YRCNGTDI YKDKESTVQVHYRMCQSCVELDPATVAGI IVTDVIATLL LALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLT CGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS ISGTTVILTCPQY PGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYL RARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQN KERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGPMRISKPHLR S IS IQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDA TLYTESDVHPSCKVTAMKCFLLELQVISLESGDAS IHDTVENLI ILANNSLSSNGNVTESGC KECEELEEKNIKEFLQSFVHIVQMFINTS* ( SEQ ID NO : 74 )

[0218] In TCRpp65ZFLGDEFL15, the corresponding component sequences are as follows, although these particular sequences or others may be utilized in this and/or other constructs:

[0219] TCRb-extracellular domain'.

MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVP NGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGI YGYTFGSGTRLTWEDLNKVFPP EVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALND SRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADATN FSLLKQAGDVEENPGP ( SEQ ID NO : 75 ) ( and that includes the P2A sequence at its C-terminus )

ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGAC CCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAA TGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCC AACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGC CCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGAT ACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCC GAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGT GTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGG AGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGAC AGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAA CCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACC GGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC ( SEQ ID NO : 7 6 )

[0220] TCRa-extracellular domain'.

MILNVEQSPQSLHVQEGDSTNFTCS FPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGR I SATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVI PNIQNPDPAVYQLR DSKSSDKSVCLFTDFDSQTNVSQSKDSDAYI TDKTVLDMRSMDFKSNSAVAWSNKSDFACAN AFNNS I I PEDTFFPSPESSEGRGSLLTCGDVEENPGP ( SEQ ID NO : 77 ) ( and that includes the T2A sequence at its C-terminus )

ATGATCCTGAACGTGGAGCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAA CTTCACCTGCAGCTTCCCCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCG CCAAGAGCCCCGAGGCCCTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGG ATCAGCGCCACCCTGAACACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCC CGAGGACAGCGCCACCTACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCG GAACCAGCCTGACCGTGATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGG GACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGT GAGCCAGAGCAAGGACAGCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCA TGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAAC GCCTTCAACAACAGCATCATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGCGCCAC CAACTTCTCCCTGCTGAAGCAGGCCGGCGACGTGGAGGAGAACCCCGGCCCC ( SEQ ID NO : 78 )

[0221] TCR5: referred to TCRCgdZFLGDEFL15, is the constant region of TCR gamma and delta, linked to full length CD3zeta, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon; and IL-15. Representative sequences are as follows:

[0222] TCR constant gamma-delta (TCRCgd)

ATGCGGTGGGCCCTACTGGTGCTTCTAGCTTTCCTGTCTCCTGCCAGTCAGGATAAACAACT TGATGCAGATGTTTCCCCCAAGCCCACTATTTTTCTTCCTTCGATTGCTGAAACAAAACTCC AGAAG G C T G GAAC AT AC CTTTGTCTTCTT GAGAAAT T T T T C C C AGAT AT TAT T AAGAT AC AT T GGCAAGAAAAGAAGAGCAACACGAT T C T GGGAT CCCAGGAGGGGAACACCAT GAAGAC TAA CGACACATACATGAAATTTAGCTGGTTAACGGTGCCAGAAGAGTCACTGGACAAAGAACACA GATGTATCGT C AGAC AT GAGAAT AAT AAAAAC G GAAT T GAT C AAGAAAT TATCTTTCCTCCA AT AAAGAC AGAT G T C AC C AC AG T G GAT C C C AAAT AC AAT T AT T C AAAG GAT G C AAAT GAT G T CAT CACAAT GGAT CCCAAAGACAAT T GGT CAAAAGAT GCAAAT GATACAC TAG T GC T GCAGC TCACAAACACCTCTGCATATTACACGTACCTCCTCCTGCTCCTCAAGAGTGTGGTCTATTTT GCCATCATCACCTGCTGTCTGCTTAGAAGAACGGCTTTCTGCTGCAATGGAGAGAAATCAGG AAGCGGAGCTACTAACTTTAGCCTGCTGAAGCAGGCTGGAGATGTGGAGGAGAACCCTGGAC CTATGATTCTTACTGTGGGCTTTAGCTTTTTGTTTTTCTACAGGGGCACGCTGTGTAGTCAG CCTCATACCAAACCATCCGTTTTTGTCATGAAAAATGGAACAAATGTCGCTTGTCTGGTGAA G GAAT T C T AC C C C AAG GAT AT AAGAAT AAAT CTCGTGTCATC C AAGAAGAT AAC AGAG T T T G ATCCTGCTATTGTCATCTCTCCCAGTGGGAAGTACAATGCTGTCAAGCTTGGTAAATATGAA GATT C AAAT T C AG T GAC AT G T T C AG T T C AAC AC GAC AAT AAAAC T G T G C AC T C C AC T GAC T T T GAAG T GAAGAC AGAT T C TAG AGAT C AC G T AAAAC C AAAG GAAAC T GAAAAC AC AAAG C AAC C T T C AAAGAG C T G C CAT AAAG C C AAAG C C AT AG T T C AT AC C GAGAAG G T GAAC AT GAT G T C C CTCACAGTGCTTGGGCTACGAATGCTGTTTGCAAAGACTGTTGCCGTCAATTTTCTCTTGAC TGCCAAGTTATTTTTCTTGTAA ( SEQ ID NO : 81 )

MRWALLVLLAFLSPASQDKQLDADVSPKPT I FLPS IAETKLQKAGTYLCLLEKFFPDI IKIH WQEKKSNT ILGSQEGNTMKTNDTYMKFSWLTVPEESLDKEHRCIVRHENNKNGIDQE I I FPP IKTDVTTVDPKYNYSKDANDVI TMDPKDNWSKDANDTLLLQLTNTSAYYTYLLLLLKSWYF AI I TCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMILTVGFS FLFFYRGTLCSQ PHTKPSVFVMKNGTNVACLVKEFYPKDIRINLVSSKKI TEFDPAIVI SPSGKYNAVKLGKYE DSNSVTCSVQHDNKTVHSTDFEVKTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMS LTVLGLRMLFAKTVAVNFLLTAKLFFL ( SEQ ID NO : 82 )

[0223] CD 3

MKWKALFTAAILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAP AYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYS E IGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPME QGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKMIGF LTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAAT I SGFLFAE IVS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLN FDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKI PIEELEDRVFVNCNTS I TWVEG TVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGI I VTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARN KEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGI TQTPYKVS I SGT TVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKP EDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGGLLLLVYYWSKNRKAKAKPVTRGAG AGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGP ( SEQ ID NO : 79 )

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCC GCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTA CGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGA AGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGT GAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT CAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGT GCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAA CAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTTGGCCCA GTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGGTACTTC T GAG T T GT GAT GCAGAAGCCAAAAATAT CACAT GGT T TAAAGAT GGGAAGAT GAT CGGC T T C CTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGGGATGTA T GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T G T C AGA ACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTCAGCATT TTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTCGAGAGC TTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATCGAGAAG ATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAGACCCTGAAC TTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAGCACAGCAC

CTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCAAGATCCCCA TCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGGGTGGAGGGC ACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAATCCTGGACCC CAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCACCGTGCAGG TGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCCGGCATCATC GTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGCCGGCCACGA GACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACCAGGTGTACC AGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGGGCCAGAAAC AAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGGCCCCATGCA GAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGGGCCAGG ACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGCGGCACC ACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAACGACAA GAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGAGCCTGA AGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGCAAGCCC GAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGAGATGGA CGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGCTGCTGC TGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGCGCCGGC GCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCCCGACTA CGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAATCGGAC CGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCC ( SEQ ID NO : 80 )

[0224] IL-15:

[0225] TCR6: also referred to TCRCabZFLGDEFL15, is the constant region of TCR alpha and beta, linked to full length CD3zeta, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon; and IL-15. Representative sequences are as follows:

[0226] TCR constant alpha-beta (TCRCab)

METLLGLLILWLQLQWVSS IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYI TDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNS I I PEDTFFPSPESSCDVKLVEKS FET DTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSGATNFSLLKQAGDVEENPGPMS IGL LCCAALSLLWAGPVNADLKNVFPPKVAVFEPSEAE I SHTQKATLVCLATGFYPDHVELSWWV NGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEW TQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSAT ILYE ILLGKATLYAVLVSALVLMAM VKRKDSRG ( SEQ ID NO : 83 )

ATGGAGACCCTCTTGGGCCTGCTTATCCTTTGGCTGCAGCTGCAATGGGTGAGCAGCATCCA GAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCC T AT T C AC C GAT T T T GAT T C T C AAAC AAAT G T G T C AC AAAG T AAG GAT T C T GAT G T G T AT AT C ACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTG GAGCAACAAAT C T GAC T T T GCAT GT GCAAACGCC T T CAACAACAGCAT TAT T CCAGAAGACA CCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACA GATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGT GGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCGGAAGCGGAGCTACTAACT TTAGCCTGCTGAAGCAGGCTGGAGATGTGGAGGAGAACCCTGGACCTATGAGCATCGGCCTC CTGTGCTGTGCAGCCTTGTCTCTCCTGTGGGCAGGTCCAGTGAATGCTGACCTGAAAAACGT GTTCCCACCCAAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGG CCACACTGGTATGCCTGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTG AATGGGAAGGAGGTGCACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGC CCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGA ACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGG ACCCAGGATAGGGCCAAACCCGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGA CTGTGGCTTCACCTCCGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGA TCTTGCTAGGGAAGGCCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATG GTCAAGAGAAAGGATTCCAGAGGCTAA ( SEQ ID NO : 84 )

[0227] CD 3:

MKWKALFTAAILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAP AYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYS E IGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPME QGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKMIGF LTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAAT I SGFLFAE IVS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLN FDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKI PIEELEDRVFVNCNTS I TWVEG TVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGI I VTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARN KEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGI TQTPYKVS I SGT TVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKP EDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGGLLLLVYYWSKNRKAKAKPVTRGAG AGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGP

( SEQ ID NO : 79 )

ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCC GCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTA CGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGA AGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGT GAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT CAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGT GCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAA CAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTTGGCCCA GTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGGTACTTC T GAG T T GT GAT GCAGAAGCCAAAAATAT CACAT GGT T TAAAGAT GGGAAGAT GAT CGGC T T C CTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGGGATGTA T GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T G T C AGA ACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTCAGCATT TTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTCGAGAGC TTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATCGAGAAG ATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAGACCCTGAAC TTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAGCACAGCAC CTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCAAGATCCCCA TCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGGGTGGAGGGC ACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAATCCTGGACCC CAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCACCGTGCAGG TGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCCGGCATCATC GTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGCCGGCCACGA GACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACCAGGTGTACC AGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGGGCCAGAAAC AAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGGCCCCATGCA GAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGGGCCAGG ACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGCGGCACC ACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAACGACAA GAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGAGCCTGA AGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGCAAGCCC GAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGAGATGGA CGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGCTGCTGC TGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGCGCCGGC GCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCCCGACTA CGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAATCGGAC CGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCC ( SEQ ID NO : 80 )

[0228] IL-15:

[0229] In some embodiments, a TCR construct comprises an NY-ESO-specific TCR and a CD8 alpha/beta co-receptor molecule. In some embodiments, such a construct can comprise a TCR alpha chain variable region signal peptide, a TCR alpha chain variable region, a TCR alpha chain constant region, a 2 A element (e.g., P2A element), a TCR beta chain variable region signal peptide, a TCR beta chain variable region, a TCR beta chain constant region, a 2A element (e.g., a E2A element), a CD8-beta polypeptide, a 2A element (e.g., a T2A element), and a CD8-alpha polypeptide. In some embodiments, a TCR construct comprising an NY-ESO- specific TCR and a CD8 alpha/beta co-receptor molecule nucleotide coding sequence is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 124. In some embodiments, a TCR construct comprising an NY-ESO-specific TCR and a CD8 alpha/beta co-receptor molecule amino acid sequence is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 125.

[0230] In some embodiments, a CD8 alpha co-receptor molecule is transcriptionally linked to any TCR molecule disclosed herein. In some embodiments, a CD8 alpha co-receptor molecule nucleotide coding sequence is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 126. In some embodiments, a CD8 beta co-receptor molecule nucleotide coding sequence is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 127. In some embodiments, a CD8 alpha co-receptor amino acid sequence is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 128. In some embodiments, a CD8 beta co-receptor amino acid sequence is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 129.

ATGGAGACCCTCTTGGGCCTGCTTATCCTTTGGCTGCAGCTGCAATGGGTGAGCAGCAAACA GGAGGTGACACAGATTCCTGCAGCTCTGAGTGTCCCAGAAGGAGAAAACTTGGTTCTCAACT GCAGTTTCACTGATAGCGCTATTTACAACCTCCAGTGGTTTAGGCAGGACCCTGGGAAAGGT C T CACAT CTCTGTTGCT TAT T GAG T CAAG T CAGAGAGAGCAAACAAG T GGAAGAC T T AAT GC CTCGCTGGATAAATCATCAGGACGTAGTACTTTATACATTGCAGCTTCTCAGCCTGGTGACT CAGCCACCTACCTCTGTGCTGTGAGGCCCCTTTATGGAGGAAGCTACATACCTACATTTGGA AGAGGAACCAGCCTTATTGTTCATCCGTATATCCAGAACCCTGACCCTGCCGTGTACCAGCT GAGAGAC T C T AAAT C C AG T GAG AAG T C T G T C T G C C T AT T C AC C GAT T T T GAT T C T C AAAC AA AT G T G T C AC AAAG T AAG GATTCTGATGTGTATAT C AC AGAC AAAAC T G T G C T AGAC AT GAG G TCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGC AAAC G C C T T C AAC AAC AG CAT TAT T C C AGAAGAC AC CTTCTTCCC C AG C C C AGAAAG T T C C T G T GAT G T CAAG C T G G T C GAGAAAAG C T T T GAAACAGAT AC GAAC C T AAAC T T T C AAAAC C T G TCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCT GCGGCTGTGGTCCAGCGGAAGCGGAGCTACTAACTTTAGCCTGCTGAAGCAGGCTGGAGATG TGGAGGAGAACCCTGGACCTATGAGCATCGGCCTCCTGTGCTGTGCAGCCTTGTCTCTCCTG TGGGCAGGTCCAGTGAATGCTGGTGTCACTCAGACCCCAAAATTCCAGGTCCTGAAGACAGG ACAGAGCATGACACTGCAGTGTGCCCAGGATATGAACCATGAATACATGTCCTGGTATCGAC AAGACCCAGGCATGGGGCTGAGGCTGATTCATTACTCAGTTGGTGCTGGTATCACTGACCAA GGAGAAGTCCCCAATGGCTACAATGTCTCCAGATCAACCACAGAGGATTTCCCGCTCAGGCT GCTGTCGGCTGCTCCCTCCCAGACATCTGTGTACTTCTGTGCCAGCAGTTACGTCGGGAACA CCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACTGGAGGACCTGAAAAACGTG TTCCCACCCAAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGC CACACTGGTATGCCTGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGA ATGGGAAGGAGGTGCACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCC CTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAA CCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGA CCCAGGATAGGGCCAAACCCGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGAC TGTGGCTTCACCTCCGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGAT CTTGCTAGGGAAGGCCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGG TCAAGAGAAAGGATTCCAGAGGCAGTGGACAGTGCACCAACTACGCCCTGCTGAAGCTGGCC GGCGACGTGGAGAGCAACCCCGGCCCCATGGCCTTGCCCGTCACTGCGCTTTTGCTCCCGCT CGCTCTTCTCCTGCATGCAGCCCGACCATCTCAATTTAGAGTTTCTCCACTCGACAGGACGT GGAACCTCGGCGAAACCGTCGAACTTAAATGTCAAGTACTTCTCTCAAATCCGACTTCTGGT TGCTCATGGCTCTTTCAGCCGAGAGGAGCAGCTGCCAGCCCCACCTTCCTGCTGTATCTCTC CCAGAACAAGCCGAAGGCCGCCGAAGGGCTCGATACTCAACGATTTAGCGGGAAGCGACTCG GGGACACGTTCGTTCTTACTCTCAGCGATTTTAGAAGAGAGAACGAGGGATATTATTTTTGT TCCGCACTCTCTAACAGCATCATGTACTTCAGTCATTTTGTACCAGTCTTTCTCCCTGCAAA ACCAACGACTACTCCAGCACCAAGACCGCCCACTCCCGCACCTACTATTGCAAGCCAACCTT TGAGTCTCCGACCAGAGGCATGCAGACCTGCTGCTGGAGGTGCAGTACATACGCGAGGGTTG GATTTTGCCTGCGATATCTATATCTGGGCCCCCTTGGCCGGCACGTGCGGGGTGCTCCTGCT GAGTCTCGTAATTACTCTTTATTGTAATCATAGAAACCGCAGAAGGGTGTGTAAGTGTCCCC GGCCTGTCGTGAAAAGCGGGGATAAGCCCAGTTTGTCTGCTCGGTACGTCGGAAGCGGTGAG GGCAGGGGAAGTCTTCTAACATGCGGGGACGTGGAGGAAAATCCCGGACCCATGAGGCCACG ACTTTGGCTGCTGCTCGCTGCACAGTTGACTGTACTGCATGGCAATAGTGTGTTGCAGCAGA CACCTGCATACATCAAGGTTCAGACAAATAAGATGGTTATGCTGAGTTGCGAGGCAAAAATT AGTTTGAGCAATATGCGGATCTACTGGTTGCGACAGAGACAGGCTCCCAGTAGTGATAGTCA CCACGAATTCCTGGCTCTTTGGGATTCCGCAAAAGGAACGATTCATGGGGAAGAAGTAGAGC AGGAGAAGATTGCGGTTTTCCGCGATGCATCTCGCTTTATCCTTAATCTTACATCCGTTAAG CCTGAGGACAGTGGGATCTATTTTTGTATGATTGTAGGGTCCCCCGAATTGACATTTGGGAA GGGTACGCAGCTCTCCGTAGTTGACTTTCTGCCCACAACGGCACAACCCACTAAGAAGTCCA CCCTGAAGAAGCGCGTCTGTCGCTTGCCCAGACCTGAAACCCAAAAGGGTCCACTCTGTTCC CCTATAACCCTGGGGTTGTTGGTGGCGGGCGTCTTGGTCCTGCTTGTTAGCTTGGGCGTAGC CATTCATCTGTGTTGCC GAAGAC G C AGAG C C C GAG T T AGAT T T AT GAAG C AAT T C T AT AAG T GA ( SEQ ID NO : 124 )

METLLGLLILWLQLQWVSSKQEVTQI PAALSVPEGENLVLNCS FTDSAIYNLQWFRQDPGKG LTSLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVRPLYGGSYI PTFG RGTSLIVHPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYI TDKTVLDMR SMDFKSNSAVAWSNKSDFACANAFNNS I I PEDTFFPSPESSCDVKLVEKS FETDTNLNFQNL SVIGFRILLLKVAGFNLLMTLRLWSSGSGATNFSLLKQAGDVEENPGPMS IGLLCCAALSLL WAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGI TDQ GEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVLEDLKNV FPPKVAVFEPSEAE I SHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPA LNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSAT ILYE ILLGKATLYAVLVSALVLMAMVKRKDSRGSGQCTNYALLKLA GDVESNPGPMALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSG CSWLFQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGYYFC SALSNS IMYFSHFVPVFLPAKPTTTPAPRPPTPAPT IASQPLSLRPEACRPAAGGAVHTRGL DFACDIYIWAPLAGTCGVLLLSLVI TLYCNHRNRRRVCKCPRPWKSGDKPSLSARYVGSGE GRGSLLTCGDVEENPGPMRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKI SLSNMRIYWLRQRQAPSSDSHHEFLALWDSAKGT IHGEEVEQEKIAVFRDASRFILNLTSVK PEDSGI YFCMIVGSPELTFGKGTQLSWDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCS PI TLGLLVAGVLVLLVSLGVAIHLCCRRRRARLRFMKQFYK* ( SEQ ID NO : 125 ) ATGAGGCCACGACTTTGGCTGCTGCTCGCTGCACAGTTGACTGTACTGCATGGCAATAGTGT GTTGCAGCAGACACCTGCATACATCAAGGTTCAGACAAATAAGATGGTTATGCTGAGTTGCG AGGCAAAAATTAGTTTGAGCAATATGCGGATCTACTGGTTGCGACAGAGACAGGCTCCCAGT AGTGATAGTCACCACGAATTCCTGGCTCTTTGGGATTCCGCAAAAGGAACGATTCATGGGGA AGAAGTAGAGCAGGAGAAGATTGCGGTTTTCCGCGATGCATCTCGCTTTATCCTTAATCTTA CATCCGTTAAGCCTGAGGACAGTGGGATCTATTTTTGTATGATTGTAGGGTCCCCCGAATTG ACATTTGGGAAGGGTACGCAGCTCTCCGTAGTTGACTTTCTGCCCACAACGGCACAACCCAC TAAGAAGTCCACCCTGAAGAAGCGCGTCTGTCGCTTGCCCAGACCTGAAACCCAAAAGGGTC CACTCTGTTCCCCTATAACCCTGGGGTTGTTGGTGGCGGGCGTCTTGGTCCTGCTTGTTAGC TTGGGCGTAGCCATTCATCTGTGTTGCCGAAGACGCAGAGCCCGACTTAGATTTATGAAGCA

ATTCTATAAGTGA ( SEQ ID NO : 12 6 )

ATGGCCTTGCCCGTCACTGCGCTTTTGCTCCCGCTCGCTCTTCTCCTGCATGCAGCCCGACC ATCTCAATTTAGAGTTTCTCCACTCGACAGGACGTGGAACCTCGGCGAAACCGTCGAACTTA AATGTCAAGTACTTCTCTCAAATCCGACTTCTGGTTGCTCATGGCTCTTTCAGCCGAGAGGA GCAGCTGCCAGCCCCACCTTCCTGCTGTATCTCTCCCAGAACAAGCCGAAGGCCGCCGAAGG GCTCGATACTCAACGATTTAGCGGGAAGCGACTCGGGGACACGTTCGTTCTTACTCTCAGCG AT T T T AGAAGAGAGAAC GAG GGATATTATTTTTGTTCCG GAG T C T C T AAC AG CAT CAT G TAG TTCAGTCATTTTGTACCAGTCTTTCTCCCTGCAAAACCAACGACTACTCCAGCACCAAGACC GCCCACTCCCGCACCTACTATTGCAAGCCAACCTTTGAGTCTCCGACCAGAGGCATGCAGAC CTGCTGCTGGAGGTGCAGTACATACGCGAGGGTTGGATTTTGCCTGCGATATCTATATCTGG GCCCCCTTGGCCGGCACGTGCGGGGTGCTCCTGCTGAGTCTCGTAATTACTCTTTATTGTAA TCATAGAAACCGCAGAAGGGTGTGTAAGTGTCCCCGGCCTGTCGTGAAAAGCGGGGATAAGC CCAGTTTGTCTGCTCGGTACGTC ( SEQ ID NO : 127 )

MRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKI SLSNMRIYWLRQRQAPS SDSHHEFLALWDSAKGT IHGEEVEQEKIAVFRDASRFILNLTSVKPEDSGIYFCMIVGSPEL TFGKGTQLSWDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCSPI TLGLLVAGVLVLLVS LGVAIHLCCRRRRARLRFMKQFYK ( SEQ ID NO : 128 )

MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPRG AAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGYYFCSALSNS IMY FSHFVPVFLPAKPTTTPAPRPPTPAPT IASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIW APLAGTCGVLLLSLVI TLYCNHRNRRRVCKCPRPWKSGDKPSLSARYV ( SEQ ID NO : 129 )

[0231] In some embodiments, a TCR construct comprises PRAME-specific TCR chains. In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises TCR alpha and TCR beta chains found in PRAME-specific TCR clone 46, clone 54, and/or clone DSK3. In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises TCR alpha and TCR beta chains that target PRAME epitopes SLLQHLIGL (SEQ ID NO: 131) and/or QLLALLPSL (SEQ ID NO: 132).

[0232] In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 133 (e.g., TCR clone 46 TCR alpha) and/or 134 (e.g., TCR clone 46 TCR beta). In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 135 (e.g., TCR clone 46 TCR alpha) and/or 136 (e.g., TCR clone 46 TCR beta).

ATGCTTCTGGAACACCTGCTGATTATCCTGTGGATGCAACTCACGTGGGTCTCCGGGCAACA ACTGAATCAAAGCCCCCAATCCATGTTTATACAGGAGGGAGAGGACGTAAGTATGAATTGCA CATCTTCATCTATCTTTAACACCTGGCTGTGGTACAAACAAGACCCCGGAGAAGGTCCTGTA CTTCTCATCGCACTTTACAAAGCAGGTGAGCTTACCAGTAACGGGAGACTCACCGCACAGTT CGGTATTACAAGAAAGGATTCCTTTCTCAACATCTCCGCTTCTATCCCTTCAGACGTCGGAA TTTATTTTTGTGCTGGTATCCCTCGAGACAATTACGGTCAAAACTTTGTATTTGGGCCTGGG ACTCGGCTGTCAGTTTTGCCGTATATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGA CAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGT CCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATG GACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGC CTTCAACAACAGCATCATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACG TGAAGCTGGTGGAGAAGTCCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGTCCGTG ATCGGCTTCAGAATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCT GTGGTCCAGC ( SEQ ID NO : 133 )

ATGGGCATTAGGCTGCTGTGCAGAGTAGCATTTTGCTTTCTGGCAGTAGGATTGGTCGATGT AAAGGTTACACAGTCCTCACGGTACTTGGTAAAGCGCACTGGTGAAAAGGTCTTTCTGGAAT GTGTACAAGATATGGATCACGAAAATATGTTTTGGTACAGGCAAGATCCCGGCCTTGGACTT AGACTGATATATTTCTCCTACGATGTTAAAATGAAGGAGAAGGGCGATATTCCAGAAGGATA TTCCGTGAGCCGCGAAAAGAAGGAGCGATTCAGTTTGATACTCGAAAGTGCCTCCACAAACC AAACCTCTATGTACCTTTGCGCGTCAACGCCGTGGCTGGCCGGTGGCAATGAACAATTCTTC GGGCCGGGTACGCGCCTCACTGTCCTGGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGC GGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTTG CGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAATGGAAAGGAGGTGCAT TCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTA TTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCA GATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAA CCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGA ATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGA CTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGT AGGGGC ( SEQ ID NO : 134 )

MLLEHLLI ILWMQLTWVSGQQLNQSPQSMFIQEGEDVSMNCTSSS I FNTWLWYKQDPGEGPV LLIALYKAGELTSNGRLTAQFGITRKDSFLNISAS IPSDVGIYFCAGIPRDNYGQNFVFGPG TRLSVLPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSM DFKSNSAVAWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSV IGFRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 135 )

MGIRLLCRVAFCFLAVGLVDVKVTQSSRYLVKRTGEKVFLECVQDMDHENMFWYRQDPGLGL RLIYFSYDVKMKEKGDIPEGYSVSREKKERFSLILESASTNQTSMYLCASTPWLAGGNEQFF GPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVH SGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAK PVTQIVSAEAWGRADCGFTSESYQQGVLSAT ILYE ILLGKATLYAVLVSALVLMAMVKRKDS RG ( SEQ ID NO : 136 )

[0233] In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 137 (e.g., TCR clone 54 TCR alpha) and/or 138 (e.g., TCR clone 54 TCR beta). In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 139 (e.g., TCR clone 54 TCR alpha) and/or 140 (e.g., TCR clone 54 TCR beta).

ATGCTGCTGCTGCTGGTGCCCGTGCTGGAAGTGATCTTCACCCTGGGCGGCACCAGAGCCCA GAGCGTGACACAGCTGGGCAGCCACGTGTCCGTGTCTGAGAGGGCCCTGGTGCTGCTGAGAT GCAACTACTCTTCTAGCGTGCCCCCCTACCTGTTTTGGTACGTGCAGTACCCCAACCAGGGG CTGCAGCTGCTCCTGAAGTACACCAGCGCCGCCACACTGGTGAAGGGCATCAACGGCTTCGA GGCCGAGTTCAAGAAGTCCGAGACAAGCTTCCACCTGACCAAGCCCAGCGCCCACATGTCTG ACGCCGCCGAGTACTTCTGTGCCGTGAGCGGCCAGACCGGCGCCAACAACCTGTTCTTCGGC ACCGGCACCCGGCTGACAGTGATCCCTTACATCCAGAACCCCGACCCCGCCGTGTACCAGCT GCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCA ACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGG AGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGC CAACGCCTTCAACAACAGCATCATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCT G C GAG G T GAAG C T G G T G GAGAAG T C C T T C GAGAC AGAC AC C AAC C T GAAC T T C C AGAAC C T G TCCGTGATCGGCTTCAGAATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCT GCGGCTGTGGTCCAGC ( SEQ ID NO : 137 )

ATGGGCTTCCGGCTGCTGTGCTGCGTGGCCTTTTGTCTGCTGGGAGCCGGACCTGTGGATAG CGGCGTGACCCAGACCCCCAAGCACCTGATCACCGCCACCGGCCAGAGAGTGACCCTGCGCT GCAGCCCTAGAAGCGGCGACCTGAGCGTGTACTGGTATCAGCAGAGCCTCGACCAGGGCCTG CAGTTCCTGATCCAGTACTACAACGGCGAGGAACGGGCCAAGGGCAACATCCTGGAACGGTT CAGCGCCCAGCAGTTCCCCGATCTGCACAGCGAGCTGAACCTGAGCAGCCTGGAACTGGGCG ACAGCGCCCTGTACTTCTGCGCCAGCGCCAGATGGGATAGAGGCGGCGAGCAGTACTTCGGC CCTGGCACCAGACTGACCGTGACCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGT TTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTTGCGA CGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAATGGAAAGGAGGTGCATTCC GGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTG CCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGAT GTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCC GTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATC ATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTC TGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGG GGC ( SEQ ID NO : 138 )

MLLLLVPVLEVI FTLGGTRAQSVTQLGSHVSVSERALVLLRCNYSSSVPPYLFWYVQYPNQG LQLLLKYTSAATLVKGINGFEAEFKKSETS FHLTKPSAHMSDAAEYFCAVSGQTGANNLFFG TGTRLTVIPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMR SMDFKSNSAVAWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNL SVIGFRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 139 )

MGFRLLCCVAFCLLGAGPVDSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGL QFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASARWDRGGEQYFG PGTRLTVTEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHS GVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSR G ( SEQ ID NO : 140 )

[0234] In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 141 (e.g., TCR clone DSK3 TCR alpha) and/or 142 (e.g., TCR clone DSK3 TCR beta). In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 143 (e.g., TCR clone DSK3 TCR alpha) and/or 144 (e g., TCR clone DSK3 TCR beta).

ATGAAGAGCCTGAGGGTACTGCTGGTGATATTGTGGCTTCAGCTTAGTTGGGTCTGGTCACA ACAAAAGGAAGTTGAGCAAAACTCAGGACCACTGAGTGTACCCGAGGGCGCTATAGCATCAC TGAACTGTACCTACTCAGATCGGGGAAGCCAATCCTTTTTCTGGTACAGACAGTATTCCGGG AAGAGTCCTGAGTTGATCATGTTTATATACTCCAATGGCGATAAGGAGGATGGACGCTTCAC CGCTCAGCTTAATAAAGCGTCACAGTATGTATCCCTCCTGATTCGGGACTCACAACCATCTG ACTCTGCAACATACCTTTGTGCCGTAAAGGACAACGCCGGGAACATGCTCACTTTTGGAGGA GGTACCCGGCTTATGGTAAAACCACATATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCG GGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACG TGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGC ATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAA CGCCTTCAACAACAGCATCATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCG AC G T GAAG C T G G T G GAGAAG T C C T T C GAGAC AGAC AC C AAC C T GAAC T T C C AGAAC C T G T C C GTGATCGGCTTCAGAATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGCG GCTGTGGTCCAGC ( SEQ ID NO : 141 )

MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSG KSPELIMFI YSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVKDNAGNMLTFGG GTRLMVKPHIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRS MDFKSNSAVAWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLS VIGFRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 142 )

ATGGGATTCCGGCTTCTTTGTTGTGTGGCATTTTGTCTGTTGGGTGCGGGTCCAGTCGATAG TGGTGTAACTCAGACACCAAAACACCTTATCACGGCAACTGGGCAACGAGTGACGCTCCGCT GTAGCCCGAGGTCCGGTGATTTGAGTGTGTACTGGTACCAGCAATCTTTGGACCAGGGCTTG CAGTTCCTCATACAGTATTACAATGGTGAAGAAAGAGCGAAGGGTAATATCCTGGAAAGATT CTCCGCACAACAGTTTCCTGATCTCCACAGCGAACTGAACCTGAGTTCTCTCGAGCTCGGGG ATAGTGCTTTGTACTTCTGCGCGTCATCCGACGGTGGCGGAGTCTATGAACAATATTTCGGC CCAGGGACTAGGCTTACGGTGACGGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGT TTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTTGCGA CGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAATGGAAAGGAGGTGCATTCC GGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTG CCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGAT GTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCC GTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATC ATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTC TGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGG GGC ( SEQ ID NO : 143 )

MGFRLLCCVAFCLLGAGPVDSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGL QFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASSDGGGVYEQYFG PGTRLTVTEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHS GVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSR G ( SEQ ID NO : 144 )

[0235] In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 145-152. In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises TCR alpha and TCR beta chains found in PRAME-specific TCR clone T116-49 and/or T402-93 and/or modified versions thereof. In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises TCR alpha and TCR beta chains that target PRAME epitope LYVDSLFFL (SEQ ID NO: 167). In some embodiments, PRAME-specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in international patent application publication WO 2022/063966 Al, which is incorporated herein by reference for the purpose described herein. In some embodiments, a TCR construct comprising PRAME-specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 153-166.

ATGGAGACACTGCTGAAGGTGCTGTCTGGCACACTGCTGTGGCAGCTGACCTGGGTCCGATC TCAGCAGCCTGTTCAGTCTCCTCAGGCCGTGATCCTGAGAGAAGGCGAGGACGCCGTGATCA ACTGCAGCAGCTCTAAGGCCCTGTACAGCGTGCACTGGTACAGACAGAAGCACGGCGAGGCC CCTGTGTTCCTGATGATCCTGCTGAAAGGCGGCGAGCAGAAGGGCCACGAGAAGATCAGCGC CAGCTTCAACGAGAAGAAGCAGCAGTCCAGCCTGTACCTGACAGCCAGCCAGCTGAGCTACA GCGGCACCTACTTTTGCGGCACAGCCAATAGCGGCGGCAGCAACTACAAGCTGACCTTCGGC AAGGGCACCCTGCTGACCGTGAATCCCAAT ( SEQ ID NO : 145 ) ATGCTGCTGATCACCTCCATGCTGGTGCTGTGGATGCAGCTGAGCCAAGTGAACGGCCAGCA AGTGATGCAGATCCCTCAGTACCAGCACGTGCAAGAAGGCGAGGACTTCACCACCTACTGCA ACAGCAGCACCACACTGAGCAACATCCAGTGGTACAAGCAGCGGCCTGGCGGACACCCTGTG TTTCTGATCCAGCTGGTCAAGTCCGGCGAAGTGAAGAAGCAGAAGCGGCTGACCTTCCAGTT CGGCGAGGCCAAGAAGAACAGCAGCCTGCACATCACCGCCACACAGACCACCGATGTGGGCA CCTACTTTTGTGCTGGCGCCCTGCCTAGAGCCGGCAGCTATCAACTGACATTCGGCAAGGGC ACCAAGCTGAGCGTGATCCCCAAC ( SEQ ID NO : 14 6 )

ATGGAGACACTGCTGAAGGTGCTGTCTGGCACACTGCTGTGGCAGCTGACCTGGGTCCGATC TCAGCAGCCTGTTCAGTCTCCTCAGGCCGTGATCCTGAGAGAAGGCGAGGACGCCGTGATCA ACTGCAGCAGCTCTAAGGCCCTGTACAGCGTGCACTGGTACAGACAGAAGCACGGCGAGGCC CCTGTGTTCCTGATGATCCTGCTGAAAGGCGGCGAGCAGAAGGGCCACGAGAAGATCAGCGC CAGCTTCAACGAGAAGAAGCAGCAGTCCAGCCTGTACCTGACAGCCAGCCAGCTGAGCTACA GCGGCACCTACTTTTGCGGCACAGCCAATAGCGGCGGCAGCAACTACAAGCTGACCTTCGGC AAGGGCACCCTGCTGACCGTGAATCCCAATATCCAGAATCCGGAGCCCGCCGTATACCAGCT GAAGGACCCTAGAAGCCAGGACAGCACCCTGTGCCTGTTCACCGACTTCGACAGCCAGATCA ACGTGCCCAAGACCATGGAAAGCGGCACCTTCATCACCGACAAGACAGTGCTGGACATGAAG GCCATGGACAGCAAGTCCAACGGCGCAATCGCCTGGTCCAACCAGACCAGCTTCACATGCCA GGACATCTTCAAAGAGACAAACGCCACATACCCCAGCAGCGACGTGCCCTGTGATGCCACCC T GAG AGAGAAG T C C T T C GAGAC AGAC AT GAAC C T GAAC T T C C AGAAT CTGTCCGTGATGGGC CTGAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAATCTGCTGATGACCCTGCGGCTGTGGTC CAGC ( SEQ ID NO : 147 )

ATGCTGCTGATCACCTCCATGCTGGTGCTGTGGATGCAGCTGAGCCAAGTGAACGGCCAGCA AGTGATGCAGATCCCTCAGTACCAGCACGTGCAAGAAGGCGAGGACTTCACCACCTACTGCA ACAGCAGCACCACACTGAGCAACATCCAGTGGTACAAGCAGCGGCCTGGCGGACACCCTGTG TTTCTGATCCAGCTGGTCAAGTCCGGCGAAGTGAAGAAGCAGAAGCGGCTGACCTTCCAGTT CGGCGAGGCCAAGAAGAACAGCAGCCTGCACATCACCGCCACACAGACCACCGATGTGGGCA CCTACTTTTGTGCTGGCGCCCTGCCTAGAGCCGGCAGCTATCAACTGACATTCGGCAAGGGC ACCAAGCTGAGCGTGATCCCCAACATCCAGAATCCGGAGCCCGCCGTATACCAGCTGAAGGA CCCTAGAAGCCAGGACAGCACCCTGTGCCTGTTCACCGACTTCGACAGCCAGATCAACGTGC CCAAGACCATGGAAAGCGGCACCTTCATCACCGACAAGACAGTGCTGGACATGAAGGCCATG GACAGCAAGTCCAACGGCGCAATCGCCTGGTCCAACCAGACCAGCTTCACATGCCAGGACAT CTTCAAAGAGACAAACGCCACATACCCCAGCAGCGACGTGCCCTGTGATGCCACCCTGACAG AGAAGTCCTTCGAGACAGACATGAACCTGAACTTCCAGAATCTGTCCGTGATGGGCCTGAGA ATCCTGCTGCTGAAGGTGGCCGGCTTCAATCTGCTGATGACCCTGCGGCTGTGGTCCAGC

( SEQ ID NO : 148 )

ATGGGCACCAGACTGTTCTTCTACGTGGCCCTGTGTCTGCTGTGGACAGGCCATGTGGATGC C G GAAT C AC AC AGAG C C C C AGAC AC AAAG T GAC C GAGAC AG G C AC C C C T G T GAC AC T GAGAT GTCACCAGACCGAGAACCATCGGTACATGTATTGGTACAGACAGGACCCCGGCCACGGCCTG AGACTGATCCACTATAGCTACGGCGTGAAGGACACCGACAAGGGCGAAGTGTCTGACGGCTA CAGCGTGTCCAGAAGCAAGACCGAGGACTTCCTGCTGACCCTGGAAAGCGCCACAAGCAGCC AGACCAGCGTGTACTTCTGCGCCATCAGCGACTACGAGGGCACCGAGGCCTTTTTTGGCCAA GGCACAAGACTGACCGTGGTG ( SEQ ID NO : 149 )

ATGCTGTGTTCTCTGCTGGCTCTGCTGCTGGGCACCTTTTTTGGCGTCAGAAGCCAGACCAT CCACCAGTGGCCTGCTACACTGGTGCAGCCTGTTGGAAGCCCTCTGAGCCTGGAATGTACCG TGGAAGGCACCAGCAATCCCAACCTGTACTGGTACAGACAGGCCGCTGGAAGAGGACTGCAG CTGCTGTTTTACAGCGTCGGCATCGGCCAGATCAGCAGCGAGGTTCCACAGAATCTGAGCGC CAGCAGACCCCAGGACAGACAGTTTATCCTGAGCAGCAAGAAGCTGCTGCTGAGCGACAGCG GCTTCTACCTGTGTGCTTGGAGCCTCGGAGCCGGCTACACCGACACACAGTATTTTGGCCCT GGCACCAGACTGACCGTGCTG ( SEQ ID NO : 150 )

ATGGGCACCAGACTGTTCTTCTACGTGGCCCTGTGTCTGCTGTGGACAGGCCATGTGGATGC C G GAAT C AC AC AGAG C C C C AGAC AC AAAG T GAC C GAGAC AG G C AC C C C T G T GAC AC T GAGAT GTCACCAGACCGAGAACCATCGGTACATGTATTGGTACAGACAGGACCCCGGCCACGGCCTG AGACTGATCCACTATAGCTACGGCGTGAAGGACACCGACAAGGGCGAAGTGTCTGACGGCTA CAGCGTGTCCAGAAGCAAGACCGAGGACTTCCTGCTGACCCTGGAAAGCGCCACAAGCAGCC AGACCAGCGTGTACTTCTGCGCCATCAGCGACTACGAGGGCACCGAGGCCTTTTTTGGCCAA GGCACAAGACTGACCGTGGTGGAAGATCTCCGGAACGTGACCCCCCCTAAAGTGACCCTGTT CGAACCCAGCAAGGCCGAGATCGCCAACAAGCAGAAAGCCACCCTCGTGTGCCTGGCCAGAG GCTTCTTCCCCGACCATGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGA GTGTCCACCGACCCTCAGGCCTACAAAGAGAGCAACTACAGCTACTGCCTGAGCAGCAGACT GCGGGTGTCCGCCACCTTCTGGCACAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTTC ACGGCCTGAGCGAAGAGGACAAGTGGCCCGAAGGCTCCCCCAAGCCCGTGACCCAGAATATC TCTGCCGAGGCCTGGGGCAGAGCCGACTGTGGAATTACCAGCGCCAGCTACCACCAGGGCGT GCTGTCTGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGG TGTCTGGCCTGGTGCTGATGGCCATGGTCAAGAAGAAGAACAGC ( SEQ ID NO : 151 )

ATGCTGTGTTCTCTGCTGGCTCTGCTGCTGGGCACCTTTTTTGGCGTCAGAAGCCAGACCAT CCACCAGTGGCCTGCTACACTGGTGCAGCCTGTTGGAAGCCCTCTGAGCCTGGAATGTACCG TGGAAGGCACCAGCAATCCCAACCTGTACTGGTACAGACAGGCCGCTGGAAGAGGACTGCAG CTGCTGTTTTACAGCGTCGGCATCGGCCAGATCAGCAGCGAGGTTCCACAGAATCTGAGCGC CAGCAGACCCCAGGACAGACAGTTTATCCTGAGCAGCAAGAAGCTGCTGCTGAGCGACAGCG GCTTCTACCTGTGTGCTTGGAGCCTCGGAGCCGGCTACACCGACACACAGTATTTTGGCCCT GGCACCAGACTGACCGTGCTGGAAGATCTCCGGAACGTGACCCCCCCTAAAGTGACCCTGTT CGAACCCAGCAAGGCCGAGATCGCCAACAAGCAGAAAGCCACCCTCGTGTGCCTGGCCAGAG GCTTCTTCCCCGACCATGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGA GTGTCCACCGACCCTCAGGCCTACAAAGAGAGCAACTACAGCTACTGCCTGAGCAGCAGACT GCGGGTGTCCGCCACCTTCTGGCACAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTTC ACGGCCTGAGCGAAGAGGACAAGTGGCCCGAAGGCTCCCCCAAGCCCGTGACCCAGAATATC TCTGCCGAGGCCTGGGGCAGAGCCGACTGTGGAATTACCAGCGCCAGCTACCACCAGGGCGT GCTGTCTGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGG TGTCTGGCCTGGTGCTGATGGCCATGGTCAAGAAGAAGAACAGC ( SEQ ID NO : 152 )

METLLKVLSGTLLWQLTWVRSQQPVQSPQAVILREGEDAVINCSSSKALYSVHWYRQKHGEA PVFLMILLKGGEQKGHEKI SAS FNEKKQQSSLYLTASQLSYSGTYFCGTANSGGSNYKLTFG KGTLLTVNPN ( SEQ ID NO : 153 )

MLLI TSMLVLWMQLSQVNGQQVMQI PQYQHVQEGEDFTTYCNSSTTLSNIQWYKQRPGGHPV FLIQLVKSGEVKKQKRLTFQFGEAKKNSSLHI TATQTTDVGTYFCAGALPRAGSYQLTFGKG TKLSVI PN ( SEQ ID NO : 154 )

IQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFI TDKTVLDMKAMDSKSNGAI AWSNQTS FTCQDI FKETNATYPSSDVPCDATLTEKS FETDMNLNFQNLSVMGLRILLLKVAG FNLLMTLRLWSS ( SEQ ID NO : 155 )

IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYI TDKTVLDMRSMDFKSNSAV AWSNKSDFACANAFNNS I I PEDTFFPSPESSCDVKLVEKS FETDTNLNFQNLSVIGFRILLL KVAG FNLLMTLRLWSS ( SEQ ID NO : 156 ) IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAV AWSNKSDFACANAFNNS I IPEDTFFPSSDVPCDVKLVEKSFETDTNLNFQNLSVIGFRILLL KVAGFNLLMTLRLWSS ( SEQ ID NO : 157 )

METLLKVLSGTLLWQLTWVRSQQPVQSPQAVILREGEDAVINCSSSKALYSVHWYRQKHGEA PVFLMILLKGGEQKGHEKISASFNEKKQQSSLYLTASQLSYSGTYFCGTANSGGSNYKLTFG KGTLLTVNPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMK AMDSKSNGAIAWSNQTSFTCQDI FKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMG LRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 158 )

MLLITSMLVLWMQLSQVNGQQVMQIPQYQHVQEGEDFTTYCNSSTTLSNIQWYKQRPGGHPV FLIQLVKSGEVKKQKRLTFQFGEAKKNSSLHITATQTTDVGTYFCAGALPRAGSYQLTFGKG TKLSVIPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAM DSKSNGAIAWSNQTSFTCQDI FKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLR ILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 159 )

MGTRLFFYVALCLLWTGHVDAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGL RLIHYSYGVKDTDKGEVSDGYSVSRSKTEDFLLTLESATSSQTSVYFCAISDYEGTEAFFGQ GTRLTW ( SEQ ID NO : 160 )

MLCSLLALLLGTFFGVRSQTIHQWPATLVQPVGSPLSLECTVEGTSNPNLYWYRQAAGRGLQ LLFYSVGIGQISSEVPQNLSASRPQDRQFILSSKKLLLSDSGFYLCAWSLGAGYTDTQYFGP GTRLTVL ( SEQ ID NO : 161 )

EDLRNVTPPKVTLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVSTDPQA YKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGR ADCGITSASYHQGVLSATILYEILLGKATLYAVLVSGLVLMAMVKKKNS ( SEQ ID NO : 162 )

DLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPL KEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEA WGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 163 )

EDLNKVFPPEVAVFEPSKAEIAHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQP LKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAE AWGRADCGITSASYHQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 164 )

MGTRLFFYVALCLLWTGHVDAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGL RLIHYSYGVKDTDKGEVSDGYSVSRSKTEDFLLTLESATSSQTSVYFCAISDYEGTEAFFGQ GTRLTWEDLRNVTPPKVTLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSG VSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNI SAEAWGRADCGITSASYHQGVLSATILYEILLGKATLYAVLVSGLVLMAMVKKKNS ( SEQ

ID NO : 165 )

MLCSLLALLLGTFFGVRSQTIHQWPATLVQPVGSPLSLECTVEGTSNPNLYWYRQAAGRGLQ LLFYSVGIGQISSEVPQNLSASRPQDRQFILSSKKLLLSDSGFYLCAWSLGAGYTDTQYFGP GTRLTVLEDLRNVTPPKVTLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSG VSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNI SAEAWGRADCGI TSASYHQGVLSAT ILYE ILLGKATLYAVLVSGLVLMAMVKKKNS ( SEQ ID NO : 1 66 )

[0236] In some embodiments, a TCR construct comprises gplOO-specific TCR chains. In some embodiments, a TCR construct comprising gplOO-specific TCR chains comprises TCR alpha and TCR beta chains found in gplOO-specific TCR clone Sp(0.01)A and/or modified versions thereof. In some embodiments, a TCR construct comprising gplOO-specific TCR chains comprises TCR alpha and TCR beta chains that target gplOO epitope KTWGQYWQV (SEQ ID NO: 168). In some embodiments, gplOO-specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in patent publication US 8,216,565 B2, which is incorporated herein by reference for the purpose described herein.

[0237] In some embodiments, a TCR construct comprising gplOO-specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 169 and/or 170. In some embodiments, a TCR construct comprising gplOO-specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 171-174.

ATGAAATCCTTGAGTGTTTCCCTAGTGGTCCTGTGGCTCCAGTTAAACTGGGTGAACAGCCA GCAGAAGGTGCAGCAGAGCCCAGAATCCCTCATTGTCCCAGAGGGAGCCATGACCTCTCTCA ACTGCACTTTCAGCGACAGTGCTTCTCAGTATTTTGCATGGTACAGACAGCATTCTGGGAAA GCCCCCAAGGCACTGATGTCCATCTTCTCCAATGGTGAAAAAGAAGAAGGCAGATTCACAAT T GAG C T C AAT AAAG C GAG TCTGCATTTCTCGC TAG AC AT C AGAGAC TGC GAG C C GAG T GAG T CTGCTCTCTACCTCTGTGCAGCCAATAACTATGCCCAGGGATTAACCTTCGGTCTTGGCACC AGAGTATCTGTGTTTCCCTACATCCAGAACCCAGAACCTGCTGTGTACCAGTTAAAAGATCC TCGGTCTCAGGACAGCACCCTCTGCCTGTTCACCGACTTTGACTCCCAAATCAATGTGCCGA AAACCAT GGAAT C T GGAACGT T CAT GAG T GACAAAAC T GT GC T GGACAT GAAAGC TAT GGAT TCCAAGAGCAATGGGGCCATTGCCTGGAGCAACCAGACAAGCTTCACCTGCCAAGATATCTT CAAAGAGACCAACGCCACCTACCCCAGTTCAGACGTTCCCTGTGATGCCACGTTGACTGAGA AAAG C T T T GAAAC AGAT AT GAAC C T AAAC T T T C AAAAC C T G T C AG T T AT G G GAC T C C GAAT C CTCCTGCTGAAAGTAGCCGGATTTAACCTGCTCATGACGCTGAGGCTGTGGTCCAGTTGA ( SEQ ID NO : 1 69 )

ATGGGCTCCAGACTCTTCTTTGTGGTTTTGATTCTCCTGTGTGCAAAACACATGGAGGCTGC AGTCACCCAAAGTCCAAGAAGCAAGGTGGCAGTAACAGGAGGAAAGGTGACATTGAGCTGTC ACCAGACTAATAACCATGACTATATGTACTGGTATCGGCAGGACACGGGGCATGGGCTGAGG CTGATCCATTACTCATATGTCGCTGACAGCACGGAGAAAGGAGATATCCCTGATGGGTACAA GGCCTCCAGACCAAGCCAAGAGAATTTCTCTCTCATTCTGGAGTTGGCTTCCCTTTCTCAGA CAGCTGTATATTTCTGTGCCAGCAGCCCTGGGGGGGGGGGGGAACAGTACTTCGGTCCCGGC ACCAGGCTCACGGTTTTAGAGGATCTGAGAAATGTGACTCCACCCAAGGTCTCCTTGTTTGA GCCATCAAAAGCAGAGATTGCAAACAAACGAAAGGCTACCCTCGTGTGCTTGGCCAGGGGCT TCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGCAAGGAGGTCCACAGTGGGGTC AGCACGGACCCTCAGGCCTACAAGGAGAGCAATTATAGCTACTGCCTGAGCAGCCGCCTGAG GGTCTCTGCTACCTTCTGGCACAATCCTCGAAACCACTTCCGCTGCCAAGTGCAGTTCCATG GGCTTTCAGAGGAGGACAAGTGGCCAGAGGGCTCACCCAAACCTGTCACACAGAACATCAGT GCAGAGGCCTGGGGCCGAGCAGACTGTGGGATTACCTCAGCATCCTATCAACAAGGGGTCTT GTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAAGCCACCCTGTATGCTGTGCTTGTCA GTACACTGGTGGTGATGGCTATGGTCAAAAGAAAGAATTCATGA ( SEQ ID NO : 170 )

MKSLSVSLWLWLQLNWVNSQQKVQQSPESLIVPEGAMTSLNCTFSDSASQYFAWYRQHSGK APKALMS I FSNGEKEEGRFTIHLNKASLHFSLHIRDSQPSDSALYLCAANNYAQGLTFGLGT RVSVFPYIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAMD SKSNGAIAWSNQTSFTCQDI FKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLRI LLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 171 )

MGSRLFFWLILLCAKHMEAAVTQSPRSKVAVTGGKVTLSCHQTNNHDYMYWYRQDTGHGLR LIHYSYVADSTEKGDIPDGYKASRPSQENFSLILELASLSQTAVYFCASSPGGGGEQYFGPG TRLTVLEDLRNVTPPKVSLFEPSKAEIANKRKATLVCLARGFFPDHVELSWWVNGKEVHSGV STDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNIS AEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLWMAMVKRKNS ( SEQ ID NO : 172 )

QQKVQQSPESLIVPEGAMTSLNCTFSDSASQYFAWYRQHSGKAPKALMS I FSNGEKEEGRFT IHLNKASLHFSLHIRDSQPSDSALYLCAANNYAQGLTFGLGTRVSVFPY ( SEQ ID NO :

173 )

EAAVTQSPRSKVAVTGGKVTLSCHQTNNHDYMYWYRQDTGHGLRLIHYSYVADSTEKGDIPD GYKASRPSQENFSLILELASLSQTAVYFCASSPGGGGEQYFGPGTRLTVL ( SEQ ID NO :

174 )

[0238] In some embodiments, a TCR construct comprises MART- 1 -specific TCR chains. In some embodiments, a TCR construct comprising MART- 1 -specific TCR chains comprises TCR alpha and TCR beta chains found in MART- 1 -specific TCR clones F4 and/or F5 and/or modified versions thereof. In some embodiments, a TCR construct comprising MART-1- specific TCR chains comprises TCR alpha and TCR beta chains that target MART-1 epitope AAGIGILTV (SEQ ID NO: 175). In some embodiments, MART- 1 -specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in patent publication US 9,128,080 B2, which is incorporated herein by reference for the purpose described herein.

[0239] In some embodiments, a TCR construct comprising MART- 1 -specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 176-179. In some embodiments, a TCR construct comprising MART- 1 -specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 180-183.

ATGTTGCTTGAACATTTATTAATAATCTTGTGGATGCAGCTGACATGGGTCAGTGGTCAACA G C T GAAT C AGAG T C C T C AAT CTATGTTTATC GAG GAAG GAGAAGAT G T C T C C AT GAAC T G C A CTTCTTCAAGCATATTTAACACCTGGCTATGGTACAAGCAGGACCCTGGGGAAGGTCCTGTC CTCTTGATAGCCTTATATAAGGCTGGTGAATTGACCTCAAATGGAAGACTGACTGCTCAGTT TGGTATAACCAGAAAGGACAGCTTCCTGAATATCTCAGCATCCATACCTAGTGATGTAGGCA TCTACTTCTGTGCTGGTGGGACCGGTAACCAGTTCTATTTTGGGACAGGGACAAGTTTGACG G T C AT T C C AAAT AT C C AGAAC C C T GAG C C T G C C G T G T AC C AG C T GAGAGAC T C T AAAT C C AG TGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGG AT T C T GAT G T G TAT AT CACAGACAAAAC T G T GC T AGACAT GAGG T C T AT GGAC T T CAAGAGC AACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAG CATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCG AGAAAAG C T T T GAAAC AGAT AC GAAC C T AAAC T T T C AAAAC C T G T C AG T GAT T G G G T T C C GA ATCCTCCTCCTGAAGGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGC ( SEQ ID NO : 17 6 )

ATGGGCACAAGGTTGTTCTTCTATGTGGCCCTTTGTCTCCTGTGGACAGGACACATGGATGC T G GAAT C AC C C AGAG C C C AAGAC AC AAG G T C AC AGAGAC AG GAAC AC C AG T GAC T C T GAGAT GTCACCAGACTGAGAACCACCGCTATATGTACTGGTATCGACAAGACCCGGGGCATGGGCTG AGGCTGATCCATTACTCATATGGTGTTAAAGATACTGACAAAGGAGAAGTCTCAGATGGCTA TAGTGTCTCTAGATCAAAGACAGAGGATTTCCTCCTCACTCTGGAGTCCGCTACCAGCTCCC AGACATCTGTGTACTTCTGTGCCATCAGTGAGGTAGGGGTTGGGCAGCCCCAGCATTTTGGT GATGGGACTCGACTCTCCATCCTAGAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGT GTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCA CAGGCTTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGT GGGGTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTG CCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCT GTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCC GTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCATGTGGCTTTACCTCGTCCTACCA GCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAGGCCACCCTGTATG CTGTGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAAGAGAAAGGATTTC ( SEQ ID NO : 177 )

ATGATGAAATCCTTGAGAGTTTTACTAGTGATCCTGTGGCTTCAGTTGAGCTGGGTTTGGAG CCAACAGAAGGAGGTGGAGCAGAATTCTGGACCCCTCAGTGTTCCAGAGGGAGCCATTGCCT CTCTCAACTGCACTTACAGTGACCGAGGTTCCCAGTCCTTCTTCTGGTACAGACAATATTCT GGGAAAAGCCCTGAGTTGATAATGTTCATATACTCCAATGGTGACAAAGAAGATGGAAGGTT TACAGCACAGCTCAATAAAGCCAGCCAGTATGTTTCTCTGCTCATCAGAGACTCCCAGCCCA GTGATTCAGCCACCTACCTCTGTGCCGTGAACTTCGGAGGAGGAAAGCTTATCTTCGGACAG GGAACGGAGTTATCTGTGAAACCCAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAG AGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATG T G T C AC AAAG T AAG GATTCTGATGTGTATAT CACAGACAAAAC T G T G C T AGACAT GAG G T C T ATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAA CGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTG AT G T C AAG C T G G T C GAGAAAAG C T T T GAAAC AGAT AC GAAC C T AAAC T T T C AAAAC C T G T C A GTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCG GCTGTGGTCCAGCTGA ( SEQ ID NO : 178 ) ATGAGAATCAGGCTCCTGTGCTGTGTGGCCTTTTCTCTCCTGTGGGCAGGTCCAGTGATTGC TGGGATCACCCAGGCACCAACATCTCAGATCCTGGCAGCAGGACGGCGCATGACACTGAGAT GTACCCAGGATATGAGACATAATGCCATGTACTGGTATAGACAAGATCTAGGACTGGGGCTA AGGCTCATCCATTATTCAAATACTGCAGGTACCACTGGCAAAGGAGAAGTCCCTGATGGTTA TAGTGTCTCCAGAGCAAACACAGATGATTTCCCCCTCACGTTGGCGTCTGCTGTACCCTCTC AGACATCTGTGTACTTCTGTGCCAGCAGCCTAAGTTTCGGCACTGAAGCTTTCTTTGGACAA GGCACCAGACTCACAGTTGTAGAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGTGTT TGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAG

GCTTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGG GTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCT GAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTC AAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCCGTC ACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCATGTGGCTTTACCTCGTCCTACCAGCA AGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAGGCCACCCTGTATGCTG

TGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAAGAGAAAGGATTTC ( SEQ ID NO : 179 )

GQQLNQSPQSMFIQEGEDVSMNCTSSS I FNTWLWYKQDPGEGPVLLIALYKAGELTSNGRLT AQFGITRKDSFLNISAS IPSDVGIYFCAGGTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDS KSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAF NNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLW SS ( SEQ ID NO : 180 )

DAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGLRLIHYSYGVKDTDKGEVSD GYSVSRSKTEDFLLTLESATSSQTSVYFCAISEVGVGQPQHFGDGTRLS ILEDLNKVFPPEV AVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSR YCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRACGFTSS YQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 181 )

QKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFT AQLNKASQYVSLLIRDSQPSDSATYLCAVNFGGGKLI FGQGTELSVKPNIQNPDPAVYQLRD SKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANA FNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRL WSS ( SEQ ID NO : 182 )

IAGITQAPTSQILAAGRRMTLRCTQDMRHNAMYWYRQDLGLGLRLIHYSNTAGTTGKGEVPD GYSVSRANTDDFPLTLASAVPSQTSVYFCASSLSFGTEAFFGQGTRLTWEDLNKVFPPEVA VFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRY CLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRACGFTSSY QQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 183 )

[0240] In some embodiments, a TCR construct comprises Tyrosinase-specific TCR chains. In some embodiments, a TCR construct comprising Tyrosinase-specific TCR chains comprises TCR alpha and TCR beta chains found in Tyrosinase-specific TCR clone TIL 13831 and/or modified versions thereof. In some embodiments, a TCR construct comprising Tyrosinasespecific TCR chains comprises TCR alpha and TCR beta chains that target Tyrosinase epitope represented by amino acids 368-376 of tyrosinase (reactive against a class I MHC (HLA-A2)- restricted epitope (368-376) of tyrosinase). In some embodiments, Tyrosinase-specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in publication Roszkowski et al, Cancer Res. 65(4): 1570-6 (2005), which is incorporated herein by reference for the purpose described herein.

[0241] In some embodiments, a TCR construct comprises MAGE- A3 -specific TCR chains. In some embodiments, a TCR construct comprising MAGE- A3 -specific TCR chains comprises TCR alpha and TCR beta chains that target amino acids 271-279 of MAGE-A3, e.g., the epitope FLWGPRALV (SEQ ID NO: 184). In some embodiments, a TCR construct comprising MAGE- A3 -specific TCR chains comprises TCR alpha and TCR beta chains that target amino acids 112-120 of MAGE-A3, e.g., the epitope KVAELVHFL (SEQ ID NO: 185). In some embodiments, MAGE- A3 -specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in international patent application publication WO 2012/054825 Al, which is incorporated herein by reference for the purpose described herein. In certain embodiments, an anti-MAGE-A3 112-120 TCR comprise an A118T substitution relative to wild type (wherein the 118 position in the alpha chain is threonine). In certain embodiments, an anti-MAGE-A3 112-120 TCR comprises an Al 18V substitution relative to wild type (wherein the 118 position in the alpha chain is valine).

[0242] In some embodiments, a TCR construct comprising MAGE- A3 -specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 186-193. In some embodiments, a TCR construct comprising MAGE- A3 -specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 194-201.

ATGGGTCCTGTCACCTGCTCAGTTCTTGTGCTCCTCCTAATGCTCAGGAGGAGCAATGGCGA TGGAGACTCCGTGACCCAGACAGAAGGCCTGGTCACTCTCACAGAAGGGTTGCCTGTGATGC TGAACTGCACCTATCAGACTATTTACTCAAATCCTTTCCTTTTCTGGTATGTGCAACATCTC AATGAATCCCCTCGGCTACTCCTGAAGAGCTTCACAGACAACAAGAGGACCGAGCACCAAGG GTTCCACGCCACTCTCCATAAGAGCAGCAGCTCCTTCCATCTGCAGAAGTCCTCAGCGCAGC TGTCAGACTCTGCCCTGTACTACTGTGCTTTCGACACAAATGCTTACAAAGTCATCTTT ( SEQ ID NO : 186 )

ATGAGAGTTAGGCTCATCTCTGCTGTGGTGCTGTGTTCCCTAGGAACAGGCCTTGTGGACAT GAAAGTAACCCAGATGCCAAGATACCTGATCAAAAGAATGGGAGAGAATGTTTTGCTGGAAT GTGGACAGGACATGAGCCATGAAACAATGTACTGGTATCGACAAGACCCTGGTCTGGGGCTA GAG CTGATTTATATCTCATACGATGTTGATAG T AAC AG C GAAG GAGACAT C C C T AAAG GAT A CAGGGTCTCACGGAAGAAGCGGGAGCATTTCTCCCTGATTCTGGATTCTGCTAAAACAAACC AGACATCTGTGTACTTCTGTGCTAGCAGTTCAACAAACACAGAAGTCTTCTTT ( SEQ ID NO : 187 )

ATGGGTCCTGTCACCTGCTCAGTTCTTGTGCTCCTCCTAATGCTCAGGAGGAGCAATGGCGA TGGAGACTCCGTGACCCAGACAGAAGGCCTGGTCACTCTCACAGAAGGGTTGCCTGTGATGC TGAACTGCACCTATCAGACTATTTACTCAAATCCTTTCCTTTTCTGGTATGTGCAACATCTC AATGAATCCCCTCGGCTACTCCTGAAGAGCTTCACAGACAACAAGAGGACCGAGCACCAAGG GTTCCACGCCACTCTCCATAAGAGCAGCAGCTCCTTCCATCTGCAGAAGTCCTCAGCGCAGC TGTCAGACTCTGCCCTGTACTACTGTGCTTTCGACACAAATGCTTACAAAGTCATCTTTGGA AAAGGGACACATCTTCATGTTCTCCCTAACATCCAGAACCCAGAACCTGCTGTGTACCAGTT AAAAGATCCTCGGTCTCAGGACAGCACCCTCTGCCTGTTCACCGACTTTGACTCCCAAATCA ATGTGCCGAAAACCATGGAATCTGGAACGTTCATCACTGACAAAACTGTGCTGGACATGAAA GCTATGGATTCCAAGAGCAATGGGGCCATTGCCTGGAGCAACCAGACAAGCTTCACCTGCCA AGAT AT C T T C AAAGAGAC C AAC AC GAG C T AC C C C AG T T C AGAC GTTCCCTGTGATGC C AC G T T GAC T GAGAAAAG C T T T GAAAC AGAT AT GAAC C T AAAC T T T C AAAAC C T G T C AG T T AT G G GA CTCCGAATCCTCCTGCTGAAAGTAGCCGGATTTAACCTGCTCATGACGCTGAGGCTGTGGTC CAGTTGA ( SEQ ID NO : 188 )

ATGAGAGTTAGGCTCATCTCTGCTGTGGTGCTGTGTTCCCTAGGAACAGGCCTTGTGGACAT GAAAGTAACCCAGATGCCAAGATACCTGATCAAAAGAATGGGAGAGAATGTTTTGCTGGAAT GTGGACAGGACATGAGCCATGAAACAATGTACTGGTATCGACAAGACCCTGGTCTGGGGCTA C AG CTGATTTATATCTCATACGATGTTGATAG T AAC AG C GAAG GAGACAT C C C T AAAG GAT A CAGGGTCTCACGGAAGAAGCGGGAGCATTTCTCCCTGATTCTGGATTCTGCTAAAACAAACC AGAC AT C T G T G T AC T T C T G T G C T AG C AG T T C AAC AAAC AC AGAAG T C T T C T T T G G T AAAG GA ACCAGACTCACAGTTGTAGAGGATCTGAGAAATGTGACTCCACCCAAGGTCTCCTTGTTTGA GCCATCAAAAGCAGAGATTGCAAACAAACAAAAGGCTACCCTCGTGTGCTTGGCCAGGGGCT TCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGCAAGGAGGTCCACAGTGGGGTC AGCACGGACCCTCAGGCCTACAAGGAGAGCAATTATAGCTACTGCCTGAGCAGCCGCCTGAG GGTCTCTGCTACCTTCTGGCACAATCCTCGCAACCACTTCCGCTGCCAAGTGCAGTTCCATG GGCTTTCAGAGGAGGACAAGTGGCCAGAGGGCTCACCCAAACCTGTCACACAGAACATCAGT GCAGAGGCCTGGGGCCGAGCAGACTGTGGGATTACCTCAGCATCCTATCAACAAGGGGTCTT GTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAAGCCACCCTGTATGCTGTGCTTGTCA GTACACTGGTGGTGATGGCTATGGTCAAAAGAAAGAACTCGTGA ( SEQ ID NO : 189 )

ATGGTCCTAGTGACCATTCTGCTGCTCAGCGCGTTCTTCTCACTGAGAGGAAACAGTGCCCA GTCCGTGGACCAGCCTGATGCTCATGTCACGCTCTCTGAAGGAGCCTCCCTGGAGCTCAGAT GCAGTTATTCATACAGTGCAGCACCTTACCTCTTCTGGTACGTGCAGTATCCTGGCCAGAGC CTCCAGTTTCTCCTCAAATACATCACAGGAGACACCGTTGTTAAAGGCACCAAGGGCTTTGA GGCCGAGTTTAGGAAGAGTAACTCCTCTTTCAACCTGAAGAAATCCCCAGCCCATTGGAGCG ACTCAGCCAAGTACTTCTGTGCACTGGAGGGCCCGGATACAGGAAACTACAAATACGTCTT

( SEQ ID NO : 190 )

ATGGGCATCCAGACCCTCTGTTGTGTGATCTTTTATGTTCTGATAGCAAATCACACAGATGC TGGAGTTACCCAGACACCCAGACATGAGGTGGCAGAGAAAGGACAAACAATAATCCTGAAGT GTGAGCCAGTTTCAGGCCACAATGACCTTTTCTGGTACAGACAGACCAAGATACAGGGACTA GAGTTGCTGAGCTACTTCCGCAGCAAGTCTCTTATGGAAGATGGTGGGGCTTTCAAGGATCG ATT C AAAG C T GAGAT G C T AAAT TCATCCTTCTC C AC T C T GAAGAT T C AAC C T AC AGAAC C C A GGGACTCAGCTGTGTATCTGTGTGCCAGCAGTTTTGGGACAGCTAGTGCAGAAACGCTGTAT TTT ( SEQ ID NO : 191 ) ATGGTCCTAGTGACCATTCTGCTGCTCAGCGCGTTCTTCTCACTGAGAGGAAACAGTGCCCA GTCCGTGGACCAGCCTGATGCTCATGTCACGCTCTCTGAAGGAGCCTCCCTGGAGCTCAGAT GCAGTTATTCATACAGTGCAGCACCTTACCTCTTCTGGTACGTGCAGTATCCTGGCCAGAGC CTCCAGTTTCTCCTCAAATACATCACAGGAGACACCGTTGTTAAAGGCACCAAGGGCTTTGA GGCCGAGTTTAGGAAGAGTAACTCCTCTTTCAACCTGAAGAAATCCCCAGCCCATTGGAGCG ACTCAGCCAAGTACTTCTGTGCACTGGAGGGCCCGGATACAGGAAACTACAAATACGTCTTT GGAGCAGGTACCAGACTGAAGGTTATAGCACACATCCAGAACCCAGAACCTGCTGTGTACCA GTTAAAAGATCCTCGGTCTCAGGACAGCACCCTCTGCCTGTTCACCGACTTTGACTCCCAAA TCAATGTGCCGAAAACCATGGAATCTGGAACGTTCATCACTGACAAAACTGTGCTGGACATG AAAGCTATGGATTCCAAGAGCAATGGGGCCATTGCCTGGAGCAACCAGACAAGCTTCACCTG C C AAGAT AT C T T C AAAGAGAC C AAC G C C AC C T AC C C C AG T T C AGAC G T T C C C T G T GAT G C C A C G T T GAC T GAGAAAAG C T T T GAAAC AGAT AT GAAC C T AAAC T T C C AAAAC C T G T C AG T T AT G GGACTCCGAATCCTCCTGCTGAAAGTAGCCGGATTTAACCTGCTCATGACGCTGAGGCTGTG GTCCAGTTGA ( SEQ ID NO : 192 )

ATGGGCATCCAGACCCTCTGTTGTGTGATCTTTTATGTTCTGATAGCAAATCACACAGATGC TGGAGTTACCCAGACACCCAGACATGAGGTGGCAGAGAAAGGACAAACAATAATCCTGAAGT GTGAGCCAGTTTCAGGCCACAATGACCTTTTCTGGTACAGACAGACCAAGATACAGGGACTA GAGTTGCTGAGCTACTTCCGCAGCAAGTCTCTTATGGAAGATGGTGGGGCTTTCAAGGATCG ATT C AAAG C T GAGAT G C T AAAT TCATCCTTCTC C AC T C T GAAGAT T C AAC C T AC AGAAC C C A GGGACTCAGCTGTGTATCTGTGTGCCAGCAGTTTTGGGACAGCTAGTGCAGAAACGCTGTAT TTTGGCTCAGGAACCAGACTGACTGTTCTCGAGGATCTGAGAAATGTGACTCCACCCAAGGT CTCCTTGTTTGAGCCATCAAAAGCAGAGATTGCAAACAAACAAAAGGCTACCCTCGTGTGCT TGGCCAGGGGCTTCTTCCCCTGACACGTGGAGCTGAGCTGGTGGGTGAATGGCAAGGAGGTC CACAGTGGGGTCAGCACGGACCCTCAGGCCTACAAGGAGAGCAATTATAGCTACTGCCTGAG CAGCCGCCTGAGGGTCTCTGCTACCTTCTGGCACAATCCTCGAAACCACTTCCGCTGTCAAG TGCAGTTCCATGGGCTTTCAGAGGAGGACAAGTGGCCAGAGGGCTCACCCAAACCTGTCACA CAGAACATCAGTGCAGAGGCCTGGGGCCGAGCAGACTGTGGAATCACTTCAGCATCCTATCA TCAGGGGGTTCTGTCTGCAACCATCCTCTATGAGATCCTACTGGGGAAGGCCACCCTATATG CTGTGCTGGTCAGTGGCCTGGTGCTGATGGCCATGGTCAAGAAAAAAAATTCCTGA ( SEQ ID NO : 193 )

MGPVTCSVLVLLLMLRRSNGDGDSVTQTEGLVTLTEGLPVMLNCTYQT IYSNPFLFWYVQHL NESPRLLLKS FTDNKRTEHQGFHATLHKSSSS FHLQKSSAQLSDSALYYCAFDTNAYKVI F ( SEQ ID NO : 194 )

MRVRLI SAWLCSLGTGLVDMKVTQMPRYLIKRMGENVLLECGQDMSHETMYWYRQDPGLGL QLIYI SYDVDSNSEGDI PKGYRVSRKKREHFSLILDSAKTNQTSVYFCASSSTNTEVF ( SEQ ID NO : 195 )

MGPVTCSVLVLLLMLRRSNGDGDSVTQTEGLVTLTEGLPVMLNCTYQT IYSNPFLFWYVQHL NESPRLLLKS FTDNKRTEHQGFHATLHKSSSS FHLQKSSAQLSDSALYYCAFDTNAYKVI FG KGTHLHVLPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFI TDKTVLDMK AMDSKSNGAIAWSNQTS FTCQDI FKETNTTYPSSDVPCDATLTEKS FETDMNLNFQNLSVMG LRILLLKVAGFNLLMTLRLWSSL ( SEQ ID NO : 196 )

MRVRLI SAWLCSLGTGLVDMKVTQMPRYLIKRMGENVLLECGQDMSHETMYWYRQDPGLGL QLIYI SYDVDSNSEGDI PKGYRVSRKKREHFSLILDSAKTNQTSVYFCASSSTNTEVFFGKG TRLTWEDLRNVTPPKVSLFEPSKAE IANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGV STDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNI S AEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLWM ( SEQ ID NO : 197 )

MVLVTILLLSAFFSLRGNSAQSVDQPDAHVTLSEGASLELRCSYSYSAAPYLFWYVQYPGQS LQFLLKYITGDTWKGTKGFEAEFRKSNSSFNLKKSPAHWSDSAKYFCALEGPDTGNYKYV ( SEQ ID NO : 198 )

MGIQTLCCVI FYVLIANHTDAGVTQTPRHEVAEKGQTI ILKCEPVSGHNDLFWYRQTKIQGL ELLSYFRSKSLMEDGGAFKDRFKAEMLNSSFSTLKIQPTEPRDSAVYLCASSFGTASAETLY ( SEQ ID NO : 199 )

MVLVTILLLSAFFSLRGNSAQSVDQPDAHVTLSEGASLELRCSYSYSAAPYLFWYVQYPGQS LQFLLKYITGDTWKGTKGFEAEFRKSNSSFNLKKSPAHWSDSAKYFCALEGPDTGNYKYVF GAGTRLKVIAHIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDM KAMDSKSNGAIAWSNQTSFTCQDI FKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVM GLRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 200 )

MGIQTLCCVI FYVLIANHTDAGVTQTPRHEVAEKGQTI ILKCEPVSGHNDLFWYRQTKIQGL ELLSYFRSKSLMEDGGAFKDRFKAEMLNSSFSTLKIQPTEPRDSAVYLCASSFGTASAETLY FGSGTRLTVLEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPHVELSWWVNGKEVH SGVSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQ NI SAEAWGRADCGI TSAS YHQGVLSAT I LYE I LLGKATLYAVLVSGLVLMAMVKKKNS ( SEQ ID NO : 201 )

[0243] In some embodiments, a TCR construct comprises MAGE-A4-specific TCR chains. In some embodiments, a TCR construct comprising MAGE-A4-specific TCR chains comprises TCR alpha and TCR beta chains that target the epitope GVYDGREHTV (SEQ ID NO: 202). In some embodiments, a TCR construct comprising MAGE-A4-specific TCR chains comprises TCR alpha and TCR beta chains that target the epitope FMNKFIYEI (SEQ ID NO: 203). In some embodiments, MAGE-A4-specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in international patent application publications WO 2017/174824 Al and WO 2021/229212 Al, each of which are incorporated herein by reference for the purpose described herein. In certain embodiments, an anti-MAGE-A4 TCR alpha chain variable domain may have an M4V or an M4L amino acid substitution. In certain embodiments, an anti-MAGE-A4 TCR beta chain variable domain may have a N10E amino acid substitution.

[0244] In some embodiments, a TCR construct comprising MAGE-A4-specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 204-205. In some embodiments, a TCR construct comprising MAGE-A4-specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 206-214.

ATGAAGAAGCACCTGACCACCTTTCTCGTGATCCTGTGGCTGTACTTCTACCGGGGCAACGG CAAGAACCAGGTGGAACAGAGCCCCCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCCCCCTTCAGCAACCTGCGGTGGTACAAGCAGGACACCGGC AGAGGCCCTGTGTCCCTGACCATCCTGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACCCTGGACGCCGATACAAAGCAGAGCAGCCTGCACATCACCGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGGCGGCACAGACAGCTGGGGCAAGCTGCAG TTTGGCGCCGGAACACAGGTGGTCGTGACCCCCGACATCCAGAACCCTGACCCTGCCGTGTA CCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGA GCAGCTGCGACGTCAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCCAGCGGCAGCCGGGCCAAGAGA ( SEQ ID NO : 204 )

ATGGCCAGCCTGCTGTTCTTCTGCGGCGCCTTCTACCTGCTGGGCACCGGCTCTATGGATGC CGACGTGACCCAGACCCCCCGGAACAGAATCACCAAGACCGGCAAGCGGATCATGCTGGAAT GCTCCCAGACCAAGGGCCACGACCGGATGTACTGGTACAGACAGGACCCTGGCCTGGGCCTG CGGCTGATCTACTACAGCTTCGACGTGAAGGACATCAACAAGGGCGAGATCAGCGACGGCTA CAGCGTGTCCAGACAGGCTCAGGCCAAGTTCAGCCTGTCCCTGGAAAGCGCCATCCCCAACC AGACCGCCCTGTACTTTTGTGCCACAAGCGGCCAGGGCGCCTACGAGGAGCAGTTCTTTGGC CCTGGCACCCGGCTGACAGTGCTGGAAGATCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGT GTTCGAGCCTTCTGAGGCCGAAATCAGCCACACCCAGAAAGCCACACTCGTGTGTCTGGCCA CCGGCTTCTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGC GGCGTGTCCACCGATCCCCAGCCTCTGAAAGAACAGCCCGCCCTGAACGACAGCCGGTACTG CCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACCACTTCAGAT GCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACAGAGCCAAGCCC GTGACACAGATCGTGTCTGCCGAAGCTTGGGGGCGCGCCGATTGTGGCTTTACCAGCGAGAG CTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGAAAGGCCACAC TGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGG GGC ( SEQ ID NO : 205 )

MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLI ILEGKNCTLQCNYTVSPFSNLRWYKQDTG RGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICWSGGTDSWGKLQ FGAGTQVWTPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLD MRSMDFKSNSAVAWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQ NLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKR ( SEQ ID NO : 206 )

MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLI ILEGKNCTLQCNYTVSPFSNLRWYKQDTG RGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICWSGGTDSWGKLQ FGAGTQVWTPD ( SEQ ID NO : 207 )

MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGL RLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFG PGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHS GVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSR G ( SEQ ID NO : 208 ) MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGL RLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFG PGTRLTVLE ( SEQ ID NO : 209 )

MKNQVEQSPQSLI ILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTIMTFSENTKSNGR YTATLDADTKQSSLHITASQLSDSASYICWSGGTDSWGKLQF ( SEQ ID NO : 210 )

MKNQVEQSPQSLI ILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTIVTFSENTKSNGR YTATLDADTKQSSLHITASQLSDSASYICWSGGTDSWGKLQF ( SEQ ID NO : 211 )

MKNQVEQSPQSLI ILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGR YTATLDADTKQSSLHITASQLSDSASYICWSGGTDSWGKLQF ( SEQ ID NO : 212 )

MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGL RLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYNEQFF ( SEQ ID NO : 213 )

MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGL RLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFF ( SEQ ID NO : 214 )

[0245] In some embodiments, a TCR construct comprises Wilms' tumor antigen (WT1) WT1 -specific TCR chains. In some embodiments, a TCR construct comprising WT1 -specific TCR chains comprises TCR alpha and TCR beta chains that target the epitope VLDFAPPGA (SEQ ID NO: 215). In some embodiments, a TCR construct comprising WTl-specific TCR chains comprises TCR alpha and TCR beta chains that target the epitope RMFPNAPYL (SEQ ID NO: 216). In some embodiments, WTl-specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in international patent application publications WO 2020/185796 Al and WO 2021/034976 Al, each of which are incorporated herein by reference for the purpose described herein. In some embodiments, a leader sequence and/or signal peptide may be removed from a TCR amino acid sequence, and percentage sequence identity may be calculated based on the TCR amino acid sequence without the leader sequence and/or signal peptide.

[0246] In some embodiments, a TCR construct comprising WTl-specific TCR chains comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 217-256. In some embodiments, a TCR construct comprising WTl-specific TCR chains comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to one or more of SEQ ID NOs: 257-291.

ATGGAGACACTGCTGGGACTACTGATTCTGTGGCTGCAACTGCAATGGGTGAGCAGCAAACA GGAGGTTACCCAGATTCCTGCTGCTCTGTCTGTTCCTGAAGGCGAGAATCTGGTGCTGAACT GCAGCTTCACAGATAGCGCCATCTACAACCTGCAGTGGTTCAGACAGGATCCTGGAAAAGGC C T GACAAGCC T GC T GC T GAT T CAGAGC T C T CAGAGAGAGCAGACAT C T GGAAGAC T GAAT GC TAGCCTGGACAAGTCTAGCGGCAGAAGCACCCTGTATATTGCCGCCTCTCAACCTGGAGATT CTGCCACATACCTGTGTGCTGTGAAGGAGACATCTGGCTCTAGACTGACCTTTGGCGAGGGA ACACAACTGACCGT GAAT CCT GAG ( SEQ ID NO : 217 )

ATGACCAGAGTTAGCCTGTTATGGGCTGTGGTGGTGAGCACATGTCTGGAATCTGGAATGGC C C AGAC AG T GAG AC AG T C T GAG CCT GAAAT G T C T G T G C AG GAAG C C GAAAC C G T TAG AC T GA GCTGCACCTACGATACAAGCGAGAACAACTACTACCTGTTCTGGTACAAGCAGCCCCCCTCT AGGCAGATGATCCTGGTGATCAGACAGGAGGCCTATAAACAGCAGAATGCCACAGAGAACCG GTTCAGCGTGAACTTCCAGAAAGCCGCCAAGAGCTTCAGCCTGAAGATCTCTGATTCTCAGC TGGGCGATACAGCCATGTACTTTTGCGCCTTCATCTACCCCAGCTACACAAGCGGCACATAC AAGTACATCTTCGGCACCGGCACAAGACTGAAGGTTCTGGCCAAC ( SEQ ID NO : 218 )

ATGGCCATGTTACTAGGAGCGAGCGTGCTGATTCTGTGGTTACAGCCTGATTGGGTGAACTC TCAGCAGAAGAACGATGATCAGCAGGTGAAGCAGAACAGCCCCTCTCTGTCTGTGCAGGAAG GCAGAATCAGCATCCTGAATTGCGATTACACCAACAGCATGTTCGACTACTTCCTGTGGTAC AAGAAGTACCCCGCCGAGGGCCCTACCTTTCTGATCAGCATCTCTAGCATCAAGGACAAGAA CGAAGATGGCAGATTCACCGTGTTCCTGAACAAGAGCGCCAAGCACCTGAGCCTGCACATTG TGCCTTCTCAACCTGGAGATTCTGCCGTGTACTTTTGTGCTGCCTCTGGAACAGGCGGAAGC TATATCCCCACATTTGGAAGAGGAACAAGCCTGATCGTGCACCCTTAC ( SEQ ID NO : 219 )

ATGGCCATGTTACTAGGAGCGAGCGTGCTGATTCTGTGGTTACAGCCTGATTGGGTGAACTC TCAGCAGAAGAACGATGATCAGCAGGTGAAGCAGAACAGCCCCTCTCTGTCTGTGCAGGAAG GCAGAATCAGCATCCTGAATTGCGATTACACCAACAGCATGTTCGACTACTTCCTGTGGTAC AAGAAGTACCCCGCCGAGGGCCCTACCTTTCTGATCAGCATCTCTAGCATCAAGGACAAGAA CGAAGATGGCAGATTCACCGTGTTCCTGAACAAGAGCGCCAAGCACCTGAGCCTGCACATTG TGCCTTCTCAACCTGGAGATTCTGCCGTGTACTTTTGTGCTGCCTCTGGCATTGGCGACTAC AAACTGAGCTTTGGAGCCGGCACAACAGTGACCGTTAGAGCCAAT ( SEQ ID NO : 220 )

ATGGTGAAGATCCGGCAGTTCCTCCTGGCTATTCTGTGGCTGCAACTGTCTTGTGTGTCTGC TGCCAAGAATGAAGTGGAGCAGTCTCCCCAGAACCTTACAGCCCAGGAAGGCGAGTTTATCA CCATCAACTGCAGCTATTCTGTGGGCATTAGCGCCCTGCATTGGCTGCAGCAACACCCTGGA GGAGGAATTGTGTCTCTGTTTATGCTGTCTTCTGGCAAGAAGAAGCACGGCCGGCTGATTGC CACCATCAACATCCAGGAGAAGCACTCTTCTCTGCACATTACAGCCTCTCATCCCAGGGATT CTGCCGTGTACATCTGTGCCGTGAGAACCAGCTACGATAAGGTGATTTTCGGACCAGGCACC TCTCTGAGCGTGATCCCCAAT ( SEQ ID NO : 221 )

ATGAAGAGCCTGAGAGTCCTGCTGGTGATTTTGTGGCTGCAGCTGTCTTGGGTTTGGTCTCA GCAGAAAGAAGTGGAGCAGAATAGCGGCCCTCTGTCTGTTCCTGAAGGCGCTATTGCTAGCC TGAATTGCACATACAGCGATAGAGGATCTCAGAGCTTCTTCTGGTACCGGCAGTACAGCGGC AAGAGCCCAGAACTGATCATGTTCATCTACAGCAATGGCGACAAGGAGGATGGCAGGTTTAC AGCCCAGCTGAACAAGGCCAGCCAGTATGTTTCTCTGCTGATCAGAGATAGCCAGCCTAGCG ATTCTGCCACCTACCTGTGTGCCGTGAACTTACTTGGAGCTACAGGATACTCTACACTGACC TTCGGCAAAGGCACCATGCTGCTGGTGAGCCCTGAT ( SEQ ID NO : 222 ) ATGTGGGGCGTTTTCCTTCTGTATGTGAGCATGAAGATGGGCGGCACAACAGGCCAGAACAT CGATCAGCCTACCGAGATGACAGCCACAGAAGGAGCTATTGTTCAGATCAACTGCACCTACC AGACAAGCGGCTTCAACGGCCTGTTCTGGTACCAGCAGCATGCTGGAGAAGCTCCTACATTT CTGAGCTACAATGTGCTGGATGGCCTGGAGGAGAAAGGCAGGTTTAGCAGCTTCCTGAGCAG GTCTAAGGGCTATTCTTATCTGCTGCTGAAGGAGCTGCAGATGAAGGATTCCGCCAGCTACC TGTGTGCCGTTAGGGGCATCAATGATTACAAGCTGAGCTTTGGAGCCGGAACAACAGTGACC GTGAGAGCCAAC ( SEQ ID NO : 223 )

ATGGAGAAGATGCTGGAGTGTGCGTTCATCGTTCTGTGGCTGCAACTTGGATGGCTGTCTGG AGAGGATCAGGTTACACAGTCTCCTGAAGCCCTGAGACTGCAAGAAGGAGAAAGCTCTAGCC TGAACTGCAGCTACACAGTGTCTGGACTGAGAGGCCTGTTCTGGTACAGACAGGATCCTGGA AAAGGCCCAGAGTTCCTGTTTACCCTGTATTCTGCCGGCGAGGAGAAGGAGAAAGAGAGACT GAAAGCTACCCTGACCAAGAAGGAGAGCTTCCTGCACATTACCGCCCCCAAACCTGAGGATT CTGCCACATATCTGTGTGCCGTGATTACCGGCTTTCAGAAGCTGGTGTTTGGCACAGGCACC AGACTGCTGGTTTCTCCCAAT ( SEQ ID NO : 224 )

ATGAGACTGGTGGCACGCGTAACTGTGTTTCTGACCTTTGGCACCATCATCGATGCCAAGAC AACCCAGCCTACAAGCATGGACTGTGCCGAGGGAAGAGCTGCTAATCTGCCATGTAATCACA GCACAATCAGCGGCAACGAGTACGTGTACTGGTACCGGCAGATCCACTCTCAAGGACCTCAG TACATCATTCATGGCCTGAAGAACAACGAGACCAACGAGATGGCCAGCCTGATCATCACCGA GGACAGGAAGTCTTCTACCCTGATTCTGCCTCATGCTACACTGAGAGATACCGCCGTGTACT ACTGCATTGCCGGAGTGGGAAGAGGCCAGAATTTCGTGTTTGGACCTGGAACAAGACTGAGC GTTCTGCCCTAT ( SEQ ID NO : 225 )

ATGGAGAAGAACCCCTTGGCAGCACCTCTGCTTATTCTGTGGTTCCACCTGGATTGTGTGAG CAGCATCCTGAATGTGGAGCAGTCTCCTCAGAGCCTGCATGTGCAAGAAGGCGATAGCACCA ATTTCACCTGCAGCTTTCCAAGCAGCAACTTCTACGCCCTGCACTGGTACAGATGGGAAACC GCCAAATCTCCTGAAGCCCTGTTTGTGATGACCCTGAATGGCGACGAGAAGAAGAAGGGCAG AATTAGCGCCACCCTGAATACCAAGGAGGGCTACAGCTACCTGTACATCAAGGGCTCTCAAC CTGAGGATTCTGCCACCTACCTTTGCGCCTTTCACCCCAATTTCGGCAACGAGAAACTGACC TTTGGAACCGGAACAAGGCTGACCATCATCCCCAAC ( SEQ ID NO : 226 )

ATGGAGAAGATGCTGGAGTGTGCGTTCATCGTTCTGTGGCTGCAACTTGGATGGCTGTCTGG AGAGGATCAGGTTACACAGTCTCCTGAAGCCCTGAGACTGCAAGAAGGAGAAAGCTCTAGCC TGAACTGCAGCTACACAGTGTCTGGACTGAGAGGCCTGTTCTGGTACAGACAGGATCCTGGA AAAGGCCCAGAGTTCCTGTTTACCCTGTATTCTGCCGGCGAGGAGAAGGAGAAAGAGAGACT GAAAGCTACCCTGACCAAGAAGGAGAGCTTCCTGCACATTACCGCCCCCAAACCTGAGGATT CTGCCACATATCTGTGTGCTGTTCAGCCTAGAGGAGATGGCTCTAGCAATACCGGCAAGCTG ATCTTTGGCCAGGGAACAACACTGCAGGTGAAGCCTGAT ( SEQ ID NO : 227 )

ATCCAGAATCCCGATCCTGCTGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGT GTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGT ACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTG GCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGA GGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCG AGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTGATCGGCTTCCGGATCCTGCTGCTG AAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGCTGA ( SEQ ID NO : 228 ) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCACATCTCTTCTCTGT TGGGTGGTTCTGGGCTTTCTGGGCACAGATCATACAGGAGCTGGAGTTAGCCAGTCTCCTAG GTATAAGGTGACCAAGAGGGGACAGGATGTGGCTCTGAGATGTGACCCTATTAGCGGACATG TGAGCCTGTACTGGTACAGACAAGCTCTGGGACAAGGACCCGAGTTTCTGACCTACTTCAAC TATGAGGCCCAGCAGGACAAATCTGGACTGCCCAACGACAGATTCAGCGCCGAAAGACCAGA AGGCTCTATTAGCACACTGACCATCCAGAGAACAGAGCAGAGGGATTCTGCCATGTACAGAT GCGCCAGCAGCTTAACAGGCTCTTACGAGCAGTACTTTGGACCTGGCACAAGACTGACAGTG ACAGAG ( SEQ ID NO : 229 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGCTGCTTCTTCTCCTCCTT CTCGGACCTGCTGGATCTGGATTAGGAGCTGTTGTGTCTCAGCACCCTTCTTGGGTGATCTG TAAAAGCGGCACAAGCGTGAAGATCGAGTGCAGAAGCCTGGACTTTCAGGCCACAACCATGT TCTGGTATAGGCAGTTCCCCAAGCAGTCTCTGATGCTGATGGCCACCTCTAATGAGGGCTCT AAGGCCACATATGAACAGGGAGTGGAGAAGGACAAGTTCCTGATCAACCACGCCTCTCTGAC CCTGTCTACCCTGACAGTTACATCTGCCCACCCTGAGGATAGCAGCTTTTACATCTGTAGCG CCACACCTGAAGCCTCTAGCCCATATGAGCAGTACTTTGGCCCTGGCACCAGATTAACAGTG ACAGAG ( SEQ ID NO : 230 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGACCTGGACTGCTTCAT TGGATGGCTCTGTGTTTGCTGGGAACAGGACATGGAGATGCTATGGTGATCCAGAACCCCAG G T AT GAG G T GAG C GAG T T T G G C AAAC GAG T GAG AC T GAG C T G T T C T C AGAC C C T GAAC C AC A ACGTGATGTACTGGTACCAGCAGAAGTCTTCTCAGGCCCCTAAGCTGCTGTTCCACTACTAC GACAAGGACTTCAACAACGAGGCCGATACCCCTGACAATTTCCAGAGCAGGAGGCCCAATAC CAGCTTCTGTTTCCTGGACATTAGAAGCCCTGGACTGGGAGATGCTGCCATGTACCTGTGTG CCACCAGCAATTTACAGGGAAGACAACCTCAGCACTTTGGCGATGGCACAAGGCTGTCTATC CTGGAG ( SEQ ID NO : 231 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGCTGAGCCCTGATCTCCCT GATTCTGCCTGGAATACCAGACTGCTGTGTCATGTGATGCTGTGTCTGCTTGGAGCCGTTTC TGTGGCTGCTGGCGTGATTCAATCTCCTAGACACCTGATCAAGGAGAAGAGAGAAACAGCCA CCCTGAAGTGCTACCCCATCCCCAGACACGATACAGTGTACTGGTATCAGCAAGGACCTGGA CAAGATCCCCAGTTCCTGATCAGCTTCTACGAGAAGATGCAGAGCGACAAAGGCAGCATCCC AGACAGATTTAGCGCCCAGCAGTTTAGCGACTATCACTCTGAGCTGAACATGAGCAGCCTGG AACTGGGCGATTCTGCTCTGTACTTCTGTGCCTCTTCTCTGAGACTGGGAAGAGAAACCCAG TACTTTGGACCCGGCACAAGACTGCTGGTTCTTGAG ( SEQ ID NO : 232 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCACAAGACTTCTCTGC TGGGTGGTGCTTGGATTTCTGGGCACAGATCATACAGGAGCTGGAGTTAGCCAGTCTCCTAG GTACAAAGTGGCCAAGAGAGGACAGGATGTGGCTCTGAGATGTGACCCTATTAGCGGACATG TGAGCCTGTTTTGGTACCAGCAAGCTCTGGGACAAGGACCCGAGTTTCTGACCTACTTCCAG AATGAAGCCCAGCTGGATAAATCTGGACTGCCTAGCGACCGGTTCTTCGCCGAAAGACCTGA AGGATCTGTTAGCACCCTGAAGATTCAGAGAACACAGCAGGAGGACTCTGCCGTGTACCTGT GTGCCTCTTCTTTAGGACAGGCCTATGAGCAGTATTTTGGACCTGGCACCAGACTGACCGTG ACAGAG ( SEQ ID NO : 233 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCACAAGACTTCTCTGC TGGGTGGCCTTTTGTCTGCTGGTGGAAGAGCTGATTGAAGCTGGAGTTGTGCAGTCTCCTAG GTACAAGATCATCGAGAAGAAGCAGCCCGTGGCCTTCTGGTGTAATCCCATTTCTGGCCACA ACACCCTGTACTGGTATCTGCAGAATCTGGGACAGGGCCCTGAACTGCTGATCAGATACGAG AACGAAGAAGCCGTGGACGATTCTCAACTGCCTAAGGACCGCTTTTCTGCCGAGAGGCTGAA AGGAGTGGATTCTACCCTGAAGATCCAACCTGCTGAACTGGGCGATTCTGCTGTGTACCTGT GCGCTTCTAGCCTGACAAGAGGAGCTGAAGCCTTTTTTGGACAGGGCACAAGACTGACAGTG GTGGAG ( SEQ ID NO : 234 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGACCTCAGCTTCTTGGA TACGTTGTGCTGTGTCTGCTTGGAGCTGGACCTCTTGAAGCTCAGGTTACCCAGAACCCCAG AT AC CT GATT AC CGT GAC AG G C AAAAAG C T GAC C G T GAC AT G T AG C C AGAAC AT GAAC C AC G AGTACATGAGCTGGTACCGGCAGGATCCTGGATTAGGCCTGAGACAGATCTACTACAGCATG AACGTGGAGGTGACCGATAAAGGCGACGTGCCTGAGGGATACAAGGTGAGCAGAAAGGAGAA GAGGAATTTCCCCCTGATCCTGGAAAGCCCAAGCCCCAATCAGACAAGCCTGTACTTTTGTG CCAGCAGCTTTTCTGGCGGCACATATGAGCAGTACTTCGGCCCTGGCACAAGACTGACAGTT ACAGAG ( SEQ ID NO : 235 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGCTGAGCCCTGATCTCCCT GATTCTGCCTGGAATACCAGACTGCTGTGTCATGTGATGCTGTGTCTGCTTGGAGCCGTTTC TGTGGCTGCTGGCGTGATTCAATCTCCTAGACACCTGATCAAGGAGAAGAGAGAAACAGCCA CCCTGAAGTGCTACCCCATCCCCAGACACGATACAGTGTACTGGTATCAGCAAGGACCTGGA CAAGATCCCCAGTTCCTGATCAGCTTCTACGAGAAGATGCAGAGCGACAAAGGCAGCATCCC AGACAGATTTAGCGCCCAGCAGTTTAGCGACTATCACTCTGAGCTGAACATGAGCAGCCTGG AACTGGGCGATTCTGCTCTGTACTTCTGTGCCAGCAGCTATAGAGGAGGCAGCACATATGAG CAGTACTTTGGCCCTGGCACAAGACTGACAGTGACAGAG ( SEQ ID NO : 236 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGAGCACCAGACTCCTTTGC TGGATGGCTTTGTGTCTGCTTGGAGCTGAGCTGTCTGAAGCTGAAGTTGCCCAGTCTCCCAG ATACAAGATCACCGAGAAATCTCAGGCTGTGGCCTTCTGGTGTGACCCTATTTCTGGACACG CCACCCTGTACTGGTATAGGCAAATTCTGGGACAAGGCCCTGAACTGCTGGTGCAATTTCAG GATGAGAGCGTGGTGGACGATTCTCAACTGCCTAAGGACAGGTTTTCTGCCGAGCGGCTGAA AGGAGTTGATAGCACCCTGAAGATCCAACCTGCTGAACTGGGCGATTCTGCTATGTACCTGT GCGCCTCTTCTCAGAGAGATAGCCCTAACGAGAAGCTGTTCTTTGGCTCTGGAACCCAGCTG TCTGTGCTGGAG ( SEQ ID NO : 237 )

CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCTGTAGACTGTTGTGT TGTGCTGTGCTGTGTCTGTTGGGAGCTGTGCCTATGGAAACAGGCGTTACCCAGACACCTAG ACATCTGGTTATGGGCATGACCAACAAGAAGAGCCTGAAGTGCGAGCAGCATCTGGGCCATA ACGCCATGTACTGGTATAAGCAGAGCGCCAAGAAACCACTGGAACTGATGTTCGTGTACAGC CTGGAGGAGAGGGTGGAGAATAATAGCGTGCCCAGCAGATTTAGCCCTGAGTGCCCAAATTC TTCTCACCTGTTCCTGCACCTGCACACATTACAGCCCGAGGATTCTGCCCTGTACCTGTGTG CTTCTTCTCAAGACCCTTACAAGCTGAGCGGCAATACCATCTACTTCGGCGAAGGCTCTTGG CTGACAGTGGTTGAA ( SEQ ID NO : 238 )

GATCTGAACAAGGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTTCTGAGGCCGAGATCTC CCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTTCCCCGACCACGTGGAAC TGTCTTGGTGGGTCAACGGCAAAGAGGTGCACTCCGGCGTGTGCACCGATCCCCAGCCTCTG AAAGAACAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCAGACTGAGAGTGTCCGC CACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCG AGAACGACGAGTGGACCCAGGACAGAGCCAAGCCCGTGACACAGATCGTGTCTGCCGAAGCC TGGGGCAGAGCCGATTGCGGCTTTACCTCCGTGTCCTATCAGCAGGGCGTGCTGAGCGCCAC AATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCTGCCCTGG TGCTGATGGCCATGGTCAAGCGGAAGGACTTC ( SEQ ID NO : 239 )

GACCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAG CCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTACCCCGACCACGTGGAAC TGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTCTGCACCGACCCCCAGCCCCTG AAAGAGCAGCCCGCCCTGAACGACAGCCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGC CACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCG AGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGTCTGCTGAGGCC TGGGGCAGAGCCGATTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCAC CATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCCGCCCTGG TGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC ( SEQ ID NO : 240 )

ATGAAATCCTTGAGAGTTTTACTAGTGATCCTGTGGCTTCAGTTGAGCTGGGTTTGGAGCCA ACAGAAGGAGGTGGAGCAGAATTCTGGACCCCTCAGTGTTCCAGAGGGAGCCATTGCCTCTC TCAACTGCACTTACAGTGACCGAGGTTCCCAGTCCTTCTTCTGGTACAGACAATATTCTGGG AAAAG C C C T GAG TTGATAATGTTCATATACTC C AAT G G T GAG AAAGAAGAT G GAAG G T T T AC AGCACAGCTCAATAAAGCCAGCCAGTATGTTTCTCTGCTCATCAGAGACTCCCAGCCCAGTG ATTCAGCCACCTACCTCTGTGCCGTGAACATAGGAAACCATGACATGCGCTTTGGAGCAGGG AC C AGAC T GAC AG T AAAAC C AAAT AT C C AGAAC C C T GAC CCTGCCGTGTAC C AG C T GAGAGA CTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGT C AC AAAG T AAG GATTCTGATGTGTATAT C AC AGAC AAAAC T G T G C T AGAC AT GAG G T C T AT G GACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGC C T T C AAC AAC AG C AT T AT T C C AGAAGAC AC C T T C T T C C C C AG C C C AGAAAG T T C C T G T GAT G T C AAG C T G G T C GAGAAAAG C T T T GAAAC AGAT AC GAAC C T AAAC T T T C AAAAC C T G T C AG T G ATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCT GTGGTCCAGCTGA ( SEQ ID NO : 241 )

ATGGAGAAAATGTTGGAGTGTGCATTCATAGTCTTGTGGCTTCAGCTTGGCTGGTTGAGTGG AGAAGACCAGGTGACGCAGAGTCCCGAGGCCCTGAGACTCCAGGAGGGAGAGAGTAGCAGTC TCAACTGCAGTTACACAGTCAGCGGTTTAAGAGGGCTGTTCTGGTATAGGCAAGATCCTGGG AAAGGCCCTGAATTCCTCTTCACCCTGTATTCAGCTGGGGAAGAAAAGGAGAAAGAAAGGCT AAAAGCCACATTAACAAAGAAGGAAAGCTTTCTGCACATCACAGCCCCTAAACCTGAAGACT CAGCCACTTATCTCTGTGCTGTGCAGACCATGGACGGTAACCAGTTCTATTTTGGGACAGGG ACAAGTTTGACGGTCATTCCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGA CTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGT C AC AAAG T AAG GATTCTGATGTGTATAT C AC AGAC AAAAC T G T G C T AGAC AT GAG G T C T AT G GACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGC C T T C AAC AAC AG CAT TAT T C C AGAAGAC AC C T T C T T C C C C AG C C C AGAAAG T T C C T G T GAT G T C AAG C T G G T C GAGAAAAG C T T T GAAAC AGAT AC GAAC C T AAAC T T T C AAAAC C T G T C AG T G ATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCT GTGGTCCAGCTGA ( SEQ ID NO : 242 )

ATGGCATGCCCTGGCTTCCTGTGGGCACTTGTGATCTCCACCTGTCTTGAATTTAGCATGGC T C AGAC AG T C AC T C AG T C T C AAC C AGAGAT G T C T G T G C AG GAG G C AGAGAC C G T GAC C C T GA GCTGCACATATGACACCAGTGAGAGTGATTATTATTTATTCTGGTACAAGCAGCCTCCCAGC AGGCAGATGATTCTCGTTATTCGCCAAGAAGCTTATAAGCAACAGAATGCAACAGAGAATCG T T T C T C T G T GAAC T T C C AGAAAG C AG C C AAAT C C T T C AG T C T CAAGAT C T C AGAC T C AC AG C TGGGGGATGCCGCGATGTATTTCTGTGCTTCCAGTCCAGGAACCTACAAATACATCTTTGGA ACAGGCACCAGGCTGAAGGTTTTAGCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCT GAGAGAC T C T AAAT C C AG T GAC AAG T C T G T C T G C C T AT T C AC C GAT T T T GAT T C T C AAAC AA AT G T G T C AC AAAG T AAG GATTCTGATGTGTATAT C AC AGAC AAAAC T G T G C T AGAC AT GAG G TCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGC AAAC G C C T T C AAC AAC AG CAT TAT T C C AGAAGAC AC CTTCTTCCC C AG C C C AGAAAG T T C C T G T GAT G T C AAG C T G G T C GAGAAAAG C T T T GAAAC AGAT AC GAAC C T AAAC T T T C AAAAC C T G TCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCT GCGGCTGTGGTCCAGCTGA ( SEQ ID NO : 243 )

ATGACACGAGTTAGCTTGCTGTGGGCAGTCGTGGTCTCCACCTGTCTTGAATCCGGCATGGC C C AGAC AG T GAG T GAG T C T C AAC C AGAGAT G T C T G T G GAG GAG G C AGAGAC T G T GAG C C T GA G T T G GAG AT AT GAG AC GAG T GAGAG TAATTATTATTTGTTCTGG T AC AAAC AG C C T C C C AG C AGGCAGATGATTCTCGTTATTCGCCAAGAAGCTTATAAGCAACAGAATGCAACGGAGAATCG T T T C T C T G T GAAC T T C C AGAAAG C AG C C AAAT C C T T C AG T C T C AAGAT C T C AGAC T C AC AG C TGGGGGACACTGCGATGTATTTCTGTGCTTTCAACCCTTGGGAGAACTATGGTCAGAATTTT GTCTTTGGTCCCGGAACCAGATTGTCCGTGCTGCCCTATATCCAGAACCCTGACCCTGCCGT GTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATT C T C AAAC AAAT G T G T C AC AAAG T AAG GATTCTGATGTGTATAT C AC AGAC AAAAC T G T G C T A GACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTT TGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAG AAAG T T C C T G T GAT G T C AAG C T G G T C GAGAAAAG C T T T GAAACAGAT AC GAAC C T AAAC T T T CAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCT CATGACGCTGCGGCTGTGGTCCAGCTGA ( SEQ ID NO : 244 )

ATGAAGAGCCTGAGAGTCCTGCTGGTGATTTTGTGGCTGCAGCTGTCTTGGGTTTGGTCTCA GCAGAAAGAAGTGGAGCAGAATAGCGGCCCTCTGTCTGTTCCTGAAGGCGCTATTGCTAGCC TGAATTGCACATACAGCGATAGAGGATCTCAGAGCTTCTTCTGGTACCGGCAGTACAGCGGC AAGAGCCCAGAACTGATCATGTTCATCTACAGCAATGGCGACAAGGAGGATGGCAGGTTTAC AGCCCAGCTGAACAAGGCCAGCCAGTATGTTTCTCTGCTGATCAGAGATAGCCAGCCTAGCG ATTCTGCCACCTACCTGTGTGCCGTGAACATCGGAAATCACGACATGAGATTTGGAGCCGGC ACAAGACTGACCGTGAAGCCCAATATCCAGAACCCTGATCCTGCTGTGTACCAGCTGCGGGA CAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGT CCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATG GACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGC CTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACG TGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTG ATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCT GTGGTCCAGCTGA ( SEQ ID NO : 245 )

ATGGAGAAGATGCTGGAGTGTGCGTTCATCGTTCTGTGGCTGCAACTTGGATGGCTGTCTGG AGAGGATCAGGTTACACAGTCTCCTGAAGCCCTGAGACTGCAAGAAGGAGAAAGCTCTAGCC TGAACTGCAGCTACACAGTGTCTGGACTGAGAGGCCTGTTCTGGTACAGACAGGATCCTGGA AAAGGCCCAGAGTTCCTGTTTACCCTGTATTCTGCCGGCGAGGAGAAGGAGAAAGAGAGACT GAAAGCTACCCTGACCAAGAAGGAGAGCTTCCTGCACATTACCGCCCCCAAACCTGAGGATT CTGCCACATATCTGTGTGCTGTGCAGACCATGGATGGCAACCAGTTCTACTTCGGCACAGGC ACATCTCTGACCGTTATCCCCAATATCCAGAACCCTGATCCTGCCGTGTACCAGCTGCGGGA CAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGT CCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATG GACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGC CTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACG TGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTG ATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCT GTGGTCCAGCTGA ( SEQ ID NO : 24 6 )

ATGGCTTGTCCTGGATTCTTATGGGCTCTGGTGATCAGCACCTGTCTGGAGTTCTCTATGGC C C AGAC AG T GAC AC AG T C T C AG C C T GAAAT G T C T G T G C AG GAAG C C GAAAC C G T GAC AC T G T CTTGCACCTACGATACAAGCGAGAGCGACTACTACCTGTTCTGGTACAAGCAGCCTCCCTCT AGGCAGATGATCCTGGTGATTAGACAGGAGGCCTACAAACAGCAGAATGCCACCGAGAACCG GTTTAGCGTGAACTTCCAGAAAGCCGCCAAGAGCTTCAGCCTGAAAATCTCTGACAGCCAGC TGGGAGATGCTGCCATGTACTTTTGTGCCAGCTCTCCAGGCACCTACAAGTACATTTTTGGC ACCGGCACCAGACTGAAGGTGCTGGCCAATATCCAGAATCCCGATCCTGCCGTGTACCAGCT GCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCA ACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGG AGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGC CAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCT GCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTC AGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCT GCGGCTGTGGTCCAGCTGA ( SEQ ID NO : 247 )

ATGACCAGAGTTAGCCTGTTATGGGCTGTGGTGGTGAGCACATGTCTGGAATCTGGAATGGC C C AGAC AG T GAG AC AG T C T GAG C C T GAAAT G T C T G T G GAG GAAG C C GAAAC C G T TAG AC T GA GCTGCACCTACGATACAAGCGAGAGCAACTACTACCTGTTCTGGTACAAGCAGCCCCCTTCT AGGCAGATGATCCTGGTGATCAGACAGGAGGCCTATAAACAGCAGAATGCCACCGAGAACCG GTTTAGCGTGAACTTCCAGAAAGCCGCCAAGAGCTTCAGCCTGAAAATCTCTGACAGCCAGC TGGGCGATACAGCCATGTACTTTTGTGCCTTCAACCCCTGGGAGAACTATGGCCAGAATTTC GTGTTCGGCCCTGGCACCAGACTGTCTGTTCTGCCTTATATCCAGAACCCCGATCCTGCTGT GTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACA GCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTG GACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTT CGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCG AGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTC CAGAACCTCAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCT GATGACCCTGCGGCTGTGGTCCAGCTGA ( SEQ ID NO : 248 )

ATGGGCTGCAGGCTGCTCTGCTGTGCGGTTCTCTGTCTCCTGGGAGCAGTTCCCATAGACAC T GAAG T T AC C C AGAC AC C AAAAC AC CTGGTCATGG GAAT GAC AAAT AAGAAG T C T T T GAAAT GTGAACAACATATGGGGCACAGGGCTATGTATTGGTACAAGCAGAAAGCTAAGAAGCCACCG GAGCTCATGTTTGTCTACAGCTATGAGAAACTCTCTATAAATGAAAGTGTGCCAAGTCGCTT CTCACCTGAATGCCCCAACAGCTCTCTCTTAAACCTTCACCTACACGCCCTGCAGCCAGAAG ACTCAGCCCTGTATCTCTGCGCCAGCAGCCAAGGGACTAGCGGGGCAGATACGCAGTATTTT GGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGAGGTCGC TGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGG CCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCAC AGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATA CTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCC GCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAA CCTGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCACCTCCGA GTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGGCCA CCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAGGATTCC AGAGGCTAG ( SEQ ID NO : 249 )

ATGAGCATCGGCCTCCTGTGCTGTGCAGCCTTGTCTCTCCTGTGGGCAGGTCCAGTGAATGC TGGTGTCACTCAGACCCCAAAATTCCAGGTCCTGAAGACAGGACAGAGCATGACACTGCAGT GTGCCCAGGATATGAACCATGAATACATGTCCTGGTATCGACAAGACCCAGGCATGGGGCTG AGGCTGATTCATTACTCAGTTGGTGCTGGTATCACTGACCAAGGAGAAGTCCCCAATGGCTA CAATGTCTCCAGATCAACCACAGAGGATTTCCCGCTCAGGCTGCTGTCGGCTGCTCCCTCCC AGACATCTGTGTACTTCTGTGCCAGCAGTTACTCTCTTTGGGACCTTCAAGAGACCCAGTAC TTCGGGCCAGGCACGCGGCTCCTGGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGAGGT CGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCC TGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTG CACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAG ATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACT TCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCC AAACCTGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCACCTC CGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGG CCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAGGAT TCCAGAGGCTAG ( SEQ ID NO : 250 )

ATGGGCACCAGCCTCCTCTGCTGGATGGCCCTGTGTCTCCTGGGGGCAGATCACGCAGATAC TGGAGTCTCCCAGGACCCCAGACACAAGATCACAAAGAGGGGACAGAATGTAACTTTCAGGT GTGATCCAATTTCTGAACACAACCGCCTTTATTGGTACCGACAGACCCTGGGGCAGGGCCCA GAG T T T C T GAG T T AC T T C CAGAAT GAAGC T CAAC T AGAAAAAT CAAGGC T GC T GAG T GAT C G GTTCTCTGCAGAGAGGCCTAAGGGATCTTTCTCCACCTTGGAGATCCAGCGCACAGAGCAGG GGGACTCGGCCATGTATCTCTGTGCCAGCAGCTTTTCAGACGGGGGGGCTACAGATACGCAG TATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGA GGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGT GCCTGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAG GTGCACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTC CAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACC ACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGG GCCAAACCTGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCAC CTCCGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGA AGGCCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAG GATTCCAGAGGCTAG ( SEQ ID NO : 251 )

ATGCTGCTGCTTCTGCTGCTTCTGGGGCCAGCAGGCTCCGGGCTTGGTGCTGTCGTCTCTCA ACATCCGAGCTGGGTTATCTGTAAGAGTGGAACCTCTGTGAAGATCGAGTGCCGTTCCCTGG ACTTTCAGGCCACAACTATGTTTTGGTATCGTCAGTTCCCGAAACAGAGTCTCATGCTGATG GCAACTTCCAATGAGGGCTCCAAGGCCACATACGAGCAAGGCGTCGAGAAGGACAAGTTTCT CATCAACCATGCAAGCCTGACCTTGTCCACTCTGACAGTGACCAGTGCCCATCCTGAAGACA GCAGCTTCTACATCTGCAGTGCTAGACCCCATTCTCTCACAGATACGCAGTATTTTGGCCCA GGCACCCGGCTGACAGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGAGGTCGCTGTGTT TGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAG GCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGG GTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCT GAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTC AAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCTGTC ACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCACCTCCGAGTCTTA CCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGGCCACCTTGT ATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAGGATTCCAGAGGC TAG ( SEQ ID NO : 252 )

ATGGGCTGTAGACTGTTGTGTTGTGCTGTGCTGTGTCTGTTGGGAGCTGTGCCTATCGATAC AGAGGTGACCCAGACCCCTAAACATCTGGTTATGGGCATGACCAACAAGAAGAGCCTGAAGT GCGAGCAGCACATGGGCCATAGGGCCATGTATTGGTATAAGCAGAAGGCCAAGAAACCTCCT GAGCTGATGTTCGTGTACAGCTACGAGAAGCTGAGCATCAACGAGAGCGTGCCCAGCAGATT TTCTCCTGAGTGCCCTAATTCTAGCCTGCTGAATCTGCACCTGCATGCTCTGCAGCCTGAGG ATTCTGCTCTGTACCTGTGTGCTTCTTCTCAGGGCACATCTGGAGCTGATACACAGTACTTC GGACCTGGCACAAGACTGACAGTGCTGGAAGACCTGAAGAACGTGTTCCCCCCAGAGGTGGC CGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCCTGG CCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCAC AGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCGGTA CTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCA GATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAG CCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAGCGA GAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCA CCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGC CGGGGC ( SEQ ID NO : 253 )

ATGTCTATCGGTCTGCTGTGCTGTGCTGCTCTTTCTCTGCTTTGGGCTGGACCTGTGAATGC TGGAGTTACACAAACCCCCAAGTTCCAAGTGCTGAAGACAGGACAGAGCATGACCCTGCAGT GTGCTCAGGACATGAATCACGAGTACATGAGCTGGTACAGACAGGATCCTGGAATGGGCCTG AGGCTGATCCACTACTCTGTTGGAGCCGGAATTACAGATCAGGGAGAAGTGCCAAATGGCTA CAACGTGAGCAGGAGCACAACCGAGGACTTCCCCTTAAGACTGTTGTCTGCTGCTCCATCTC AGACAAGCGTGTACTTTTGCGCCAGCTCCTACTCTCTGTGGGATCTGCAGGAAACCCAGTAC TTTGGACCAGGCACAAGACTGTTAGTGCTGGAGGACCTGAAGAACGTGTTCCCCCCAGAGGT GGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCC TGGCCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTG CACAGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCG GTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACT TCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCC AAGCCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAG CGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGG CCACCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGAC AGCCGGGGC ( SEQ ID NO : 254 )

ATGGGCACATCTCTTCTCTGCTGGATGGCTCTTTGTCTGCTTGGAGCCGATCATGCCGATAC AGGAGTTAGCCAGGATCCTAGACACAAGATCACCAAGAGAGGCCAGAATGTGACCTTCCGGT GCGATCCTATCTCTGAGCACAACAGGCTGTACTGGTACAGACAAACACTGGGACAAGGACCT GAG T T C C T GAG C T AC T T C C AGAAC GAAG C C GAG C T G GAGAAG T C T AGAC T T C T GAG C GAG AG ATTTAGCGCCGAGAGACCTAAAGGCAGCTTTAGCACCCTGGAGATCCAGAGAACAGAACAGG GCGATTCTGCCATGTACCTGTGTGCTAGCAGCTTTTCTGATGGAGGCGCCACCGATACACAG TATTTCGGACCTGGCACAAGACTGACAGTGCTGGAGGACCTGAAGAACGTGTTCCCCCCAGA GGTGGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGT GCCTGGCCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAG GTGCACAGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAG CCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACC ACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGG GCCAAGCCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCAC CAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCA AGGCCACCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAG GACAGCCGGGGC ( SEQ ID NO : 255 )

ATGCTGCTTCTTCTCCTCCTTCTCGGACCTGCTGGATCTGGATTAGGAGCTGTTGTGTCTCA GCACCCTTCTTGGGTGATCTGTAAAAGCGGCACAAGCGTGAAGATCGAGTGCAGAAGCCTGG ACTTTCAGGCCACAACCATGTTCTGGTATAGGCAGTTCCCCAAGCAGTCTCTGATGCTGATG GCCACCTCTAATGAGGGCTCTAAGGCCACATATGAACAGGGAGTGGAGAAGGACAAGTTCCT GATCAACCACGCCTCTCTGACCCTGTCTACCCTGACAGTTACATCTGCCCACCCTGAGGATA GCAGCTTTTACATCTGTAGCGCCAGACCTCACAGCCTGACCGATACACAGTACTTTGGCCCT GGCACAAGACTGACAGTGTTAGAAGACCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTT CGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCG GCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGC GTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCGGTACTGTCT GAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCC

AGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTG ACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAGCGAGAGCTA CCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGT ACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC ( SEQ ID NO : 256 )

METLLGLLILWLQLQWVSSKQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKG LTSLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVKETSGSRLTFGEG TQLTVNP ( SEQ ID NO : 257 )

MTRVSLLWAVWSTCLESGMAQTVTQSQPEMSVQEAETVTLSCTYDTSENNYYLFWYKQPPS RQMILVIRQEAYKQQNATENRFSVNFQKAAKSFSLKISDSQLGDTAMYFCAFIYPSYTSGTY KYI FGTGTRLKVLAN ( SEQ ID NO : 258 )

MAMLLGASVLILWLQPDWVNSQQKNDDQQVKQNSPSLSVQEGRIS ILNCDYTNSMFDYFLWY KKYPAEGPTFLIS ISS IKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAASGTGGS YIPTFGRGTSLIVHPY ( SEQ ID NO : 259 )

MAMLLGASVLILWLQPDWVNSQQKNDDQQVKQNSPSLSVQEGRIS ILNCDYTNSMFDYFLWY KKYPAEGPTFLIS ISS IKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAASGIGDY KLSFGAGTTVTVRAN ( SEQ ID NO : 260 )

MVKIRQFLLAILWLQLSCVSAAKNEVEQSPQNLTAQEGEFITINCSYSVGISALHWLQQHPG GGIVSLFMLSSGKKKHGRLIATINIQEKHSSLHITASHPRDSAVYICAVRTSYDKVI FGPGT SLSVIPN ( SEQ ID NO : 261 )

MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSG KSPELIMEI YSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVNLLGATGYSTLT FGKGTMLLVSP ( SEQ ID NO : 262 )

MWGVFLLYVSMKMGGTTGQNIDQPTEMTATEGAIVQINCTYQTSGFNGLFWYQQHAGEAPTF LSYNVLDGLEEKGRFSSFLSRSKGYSYLLLKELQMKDSASYLCAVRGINDYKLSFGAGTTVT VRAN ( SEQ ID NO : 263 )

MEKMLECAFIVLWLQLGWLSGEDQVTQSPEALRLQEGESSSLNCSYTVSGLRGLFWYRQDPG KGPEFLFTLYSAGEEKEKERLKATLTKKESFLHITAPKPEDSATYLCAVITGFQKLVFGTGT RLLVSPN ( SEQ ID NO : 264 )

MRLVARVTVFLTFGTI IDAKTTQPTSMDCAEGRAANLPCNHSTISGNEYVYWYRQIHSQGPQ YI IHGLKNNETNEMASLI ITEDRKSSTLILPHATLRDTAVYYCIAGVGRGQNFVFGPGTRLS VLPY ( SEQ ID NO : 265 )

MEKNPLAAPLLILWFHLDCVSS ILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWET AKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCAFHPNFGNEKLT FGTGTRLTI IPN ( SEQ ID NO : 266 ) MEKMLECAFIVLWLQLGWLSGEDQVTQSPEALRLQEGESSSLNCSYTVSGLRGLFWYRQDPG KGPEFLFTLYSAGEEKEKERLKATLTKKESFLHITAPKPEDSATYLCAVQPRGDGSSNTGKL I FGQGTTLQVKP ( SEQ ID NO : 267 )

IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAV AWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLL KVAGFNLLMTLRLWSS ( SEQ ID NO : 268 )

MGTSLLCWWLGFLGTDHTGAGVSQSPRYKVTKRGQDVALRCDPISGHVSLYWYRQALGQGP EFLTYFNYEAQQDKSGLPNDRFSAERPEGS ISTLTIQRTEQRDSAMYRCASSLTGSYEQYFG PGTRLTVTE ( SEQ ID NO : 269 )

MLLLLLLLGPAGSGLGAWSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLM ATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSATPEASSPYEQYFG PGTRLTVTE ( SEQ ID NO : 270 )

MGPGLLHWMALCLLGTGHGDAMVIQNPRYQVTQFGKPVTLSCSQTLNHNVMYWYQQKSSQAP KLLFHYYDKDFNNEADTPDNFQSRRPNTSFCFLDIRSPGLGDAAMYLCATSNLQGRQPQHFG DGTRLS ILE ( SEQ ID NO : 271 )

MLSPDLPDSAWNTRLLCHVMLCLLGAVSVAAGVIQSPRHLIKEKRETATLKCYPIPRHDTVY WYQQGPGQDPQFLISFYEKMQSDKGS IPDRFSAQQFSDYHSELNMSSLELGDSALYFCASSL RLGRETQYFGPGTRLLVLE ( SEQ ID NO : 272 )

MGTRLLCWWLGFLGTDHTGAGVSQSPRYKVAKRGQDVALRCDPISGHVSLFWYQQALGQGP EFLTYFQNEAQLDKSGLPSDRFFAERPEGSVSTLKIQRTQQEDSAVYLCASSLGQAYEQYFG PGTRLTVTE ( SEQ ID NO : 273 )

MGTRLLCWVAFCLLVEELIEAGWQSPRYKI IEKKQPVAFWCNPISGHNTLYWYLQNLGQGP ELLIRYENEEAVDDSQLPKDRFSAERLKGVDSTLKIQPAELGDSAVYLCASSLTRGAEAFFG QGTRLTWE ( SEQ ID NO : 274 )

MSNQVLCCWLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGL RLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASSRDREQESPLHF GNGTRLTVTE ( SEQ ID NO : 275 )

MGPQLLGYWLCLLGAGPLEAQVTQNPRYLITVTGKKLTVTCSQNMNHEYMSWYRQDPGLGL RQIYYSMNVEVTDKGDVPEGYKVSRKEKRNFPLILESPSPNQTSLYFCASSFSGGTYEQYFG PGTRLTVTE ( SEQ ID NO : 276 )

MLSPDLPDSAWNTRLLCHVMLCLLGAVSVAAGVIQSPRHLIKEKRETATLKCYPIPRHDTVY WYQQGPGQDPQFLISFYEKMQSDKGS IPDRFSAQQFSDYHSELNMSSLELGDSALYFCASSY RGGSTYEQYFGPGTRLTVTE ( SEQ ID NO : 277 )

MSTRLLCWMALCLLGAELSEAEVAQSPRYKITEKSQAVAFWCDPISGHATLYWYRQILGQGP ELLVQFQDESWDDSQLPKDRFSAERLKGVDSTLKIQPAELGDSAMYLCASSQRDSPNEKLF FGSGTQLSVLE ( SEQ ID NO : 278 )

MGCRLLCCAVLCLLGAVPMETGVTQTPRHLVMGMTNKKSLKCEQHLGHNAMYWYKQSAKKPL ELMFVYSLEERVENNSVPSRFSPECPNSSHLFLHLHTLQPEDSALYLCASSQDPYKLSGNTI YFGEGSWLTWE ( SEQ ID NO : 279 ) DLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVCTDPQPL KEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEA WGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 280 )

DLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPL KEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEA WGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 281 )

MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSG KSPELIMEI YSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVNIGNHDMRFGAG TRLTVKPN ( SEQ ID NO : 282 )

MEKMLECAFIVLWLQLGWLSGEDQVTQSPEALRLQEGESSSLNCSYTVSGLRGLFWYRQDPG KGPEFLFTLYSAGEEKEKERLKATLTKKESFLHITAPKPEDSATYLCAVQTMDGNQFYFGTG TSLTVIPN ( SEQ ID NO : 283 )

MACPGFLWALVISTCLEFSMAQTVTQSQPEMSVQEAETVTLSCTYDTSESDYYLFWYKQPPS RQMILVIRQEAYKQQNATENRFSVNFQKAAKSFSLKISDSQLGDAAMYFCASSPGTYKYI FG TGTRLKVLAN ( SEQ ID NO : 284 )

MTRVSLLWAVWSTCLESGMAQTVTQSQPEMSVQEAETVTLSCTYDTSESNYYLFWYKQPPS RQMILVIRQEAYKQQNATENRFSVNFQKAAKSFSLKISDSQLGDTAMYFCAFNPWENYGQNF VFGPGTRLSVLPY ( SEQ ID NO : 285 )

IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAV AWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLL KVAGFNLLMTLRLWSS ( SEQ ID NO : 286 )

MGCRLLCCAVLCLLGAVPIDTEVTQTPKHLVMGMTNKKSLKCEQHMGHRAMYWYKQKAKKPP ELMFVYSYEKLS INESVPSRFSPECPNSSLLNLHLHALQPEDSALYLCASSQGTSGADTQYF GPGTRLTVLE ( SEQ ID NO : 287 )

MS IGLLCCAALSLLWAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGL RLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYSLWDLQETQY FGPGTRLLVLE ( SEQ ID NO : 288 )

MGTSLLCWMALCLLGADHADTGVSQDPRHKITKRGQNVTFRCDPISEHNRLYWYRQTLGQGP EFLTYFQNEAQLEKSRLLSDRFSAERPKGSFSTLEIQRTEQGDSAMYLCASSFSDGGATDTQ YFGPGTRLTVLE ( SEQ ID NO : 289 )

MLLLLLLLGPAGSGLGAWSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLM ATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSARPHSLTDTQYFGP GTRLTVLE ( SEQ ID NO : 290 )

DLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPL KEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEA WGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 291 ) [0247] In some embodiments, a TCR construct comprises Human papilloma virus (HPV)- specific TCR chains. In some embodiments, a TCR construct comprising an HPV-specific TCR chains comprises TCR alpha and TCR beta chains that target the HPV 18 E6 protein, and/or HPV 18 E7 protein. In some embodiments, an HPV 18 E6 epitope is amino acids 121-135 and/or amino acids 77-91 of the HPV 18 E6 protein. In some embodiments, a TCR construct comprising an HPV-specific TCR chains comprises TCR alpha and TCR beta chains that target the HPV 18 E7 protein. In some embodiments, an HPV 18 E7 epitope is amino acids 11-19. In some embodiments, HPV-specific TCR sequences, TCR variable domain sequences, CDR sequences, and/or TCR constant domain sequences, are described in international patent application publications WO 2015/009604 Al, which is incorporated herein by reference for the purpose described herein.

[0248] In some embodiments, a TCR is an invariant natural killer T cell TCR (iTCR). In some embodiments, an iTCR complex comprises an alpha (a) chain iTCR (iTCRa). In some embodiments, a construct encoding an iTCRa comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 292 (e.g., iTCRa associated with Va24-Jal8 in humans). In some embodiments, an iTCRa comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 293 (e.g., iTCRa associated with Va24-Jal8 in humans).

ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGC ( SEQ ID NO : 292 )

MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLI ILEGKNCTLQCNYTVSPFSNLRWYKQDTG RGPVSLTIMTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICWSDRGSTLGRLY FGRGTQLTVWPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLD MRSMDFKSNSAVAWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQ NLSVIGFRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 293 )

[0249] In some embodiments, an iTCR complex comprises a beta (P) chain iTCR (iTCRP). In some embodiments, a construct encoding an iTCRP comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 294, 296, or 298. In some embodiments, an iTCRP comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 295, 297, or 299.

ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGC ( SEQ ID NO : 294 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSELRALGPSSYN SPLHFGNGTRLTVTDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNG KEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQ DRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVK RKDF ( SEQ ID NO : 295 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCTCTAGTGAAGGCGGAGGACTCAAGTTGGCGAAAAACATC CAATACTTTGGTGCTGGCACCCGGCTTTCTGTCCTGGACCTGAACAAGGTGTTCCCTCCAGA GGTGGCCGTGTTCGAGCCTTCTGAGGCCGAGATCAGCCACACACAGAAAGCCACACTCGTGT GTCTGGCCACCGGCTTTTTCCCCGATCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAG GTGCACAGCGGCGTCtGCACAGATCCCCAGCCTCTGAAAGAACAGCCCGCTCTGAACGACAG CCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACC ACTTCAGATGCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGATAGA GCCAAGCCTGTGACACAGATCGTGTCTGCCGAAGCCTGGGGCAGAGCCGATTGTGGCTTTAC CAGCGTGTCATACCAGCAGGGCGTGCTGTCTGCCACCATCCTGTATGAGATTCTGCTGGGCA AAGCCACTCTGTACGCCGTGCTGGTGTCTGCCCTTGTGCTGATGGCCATGGTCAAGAGAAAG GACTTC ( SEQ ID NO : 296 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSEGGGLKLAKNI QYFGAGTRLSVLDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKE VHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDR AKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRK DE ( SEQ ID NO : 297 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCTCTAGTGAATTCGCCAGCTCCGTGCGAGGGAATACTATC TACTTTGGTGAGGGATCTTGGCTTACGGTAGTAGACCTGAACAAGGTGTTCCCTCCAGAGGT GGCCGTGTTCGAGCCTTCTGAGGCCGAGATCAGCCACACACAGAAAGCCACACTCGTGTGTC TGGCCACCGGCTTTTTCCCCGATCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTG CACAGCGGCGTCtGCACAGATCCCCAGCCTCTGAAAGAACAGCCCGCTCTGAACGACAGCCG GTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACCACT TCAGATGCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGATAGAGCC AAGCCTGTGACACAGATCGTGTCTGCCGAAGCCTGGGGCAGAGCCGATTGTGGCTTTACCAG CGTGTCATACCAGCAGGGCGTGCTGTCTGCCACCATCCTGTATGAGATTCTGCTGGGCAAAG CCACTCTGTACGCCGTGCTGGTGTCTGCCCTTGTGCTGATGGCCATGGTCAAGAGAAAGGAC TTC ( SEQ ID NO : 298 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSEFASSVRGNTI YFGEGSWLTWDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEV HSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRA KPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F ( SEQ ID NO : 299 )

[0250] In some embodiments, an NK cell comprises an iTCR complex beta (P) chain iTCR (iTCRP). In some embodiments, a construct encoding an iTCRp comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 300, 302, 304, 306, 308, 310, 312, or 314. In some embodiments, an iTCRp comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 301, 303, 305, 307, 309, 311, 313, or 315.

[0251] In some embodiments, a construct encoding an iTCRp comprises a nucleotide coding sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 300, 306, 310, or 312. In some embodiments, an iTCRP comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 301, 307, 311, or 313.

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCA GGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTT TGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGG GATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGA GTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCT TTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTC AAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTG ACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATA CCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGT ACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC

( SEQ ID NO : 300 ; iTCRp clone 3 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCATGQGAQDTQYFGP GTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSG VCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 301 ; 1TCR[3 clone 3 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGATGGGGTGGGGAGCAATCAGCCCCAGCATTTT GGTGATGGGACTCGACTCTCCATCCTAGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGC GGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcG CGACGGGATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCAT TCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTA TTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCA GATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAA CCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgt gTCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGA CTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 302 ; 1TCR[3 clone 18 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSDGVGSNQPQHF GDGTRLS ILEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVH SGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAK PVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 303 ; iTCRp clone 18 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGAGGGGGCTGGAAACACCATATATTTTGGAGAG GGAAGTTGGCTCACTGTTGTAGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGCGGTTTT TGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGG GATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGA GTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCT TTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTC AAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTG ACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgtgTCATA CCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGT ACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 304 ; 1TCR[3 clone 24 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSEGAGNTIYFGE GSWLTWEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSG VCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 305 ; 1TCR[3 clone 24 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGACAGGGATAGCAATCAGCCCCAGCATTTTGGT GATGGGACTCGACTCTCCATCCTAGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGCGGT TTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGA CGGGATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCC GGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTG CCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGAT GTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCC GTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgtgTC ATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTC TGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 306 ; 1TCR[3 clone 51 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSDRDSNQPQHFG DGTRLS ILEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHS GVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF

( SEQ ID NO : 307 ; 1TCR[3 clone 51 ) ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCGTGGGTCCGGTACCCTCCTACAATGAGCAGTTCTTC GGGCCAGGGACACGGCTCACCGTGCTAGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGC GGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcG CGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCAT TCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTA TTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCA GATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAA CCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGA ATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGA CTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGT AGGGGC ( SEQ ID NO : 308 ; iTCRp clone 56 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASVGPVPSYNEQFF GPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVH SGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAK PVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDS RG ( SEQ ID NO : 309 ; 1TCR[3 clone 56 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGGGGTGACTAGCGCCTCCTACAATGAGCAGTTC TTCGGGCCAGGGACACGGCTCACCGTGCTAGAGGACCTCAAGAATGTGTTTCCGCCCGAAGT CGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCC TcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTG CATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAG GTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACT TCAGATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCT AAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATC AGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAG CGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGAT AGTAGGGGC ( SEQ ID NO : 310 ; 1TCRP clone 76 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSGVTSASYNEQF FGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEV HSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRA KPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG ( SEQ ID NO : 311 ; 1TCRP clone 76 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGGAGGAGGGGAGGAGACCCAGTACTTCGGGCCA GGCACGCGGCTCCTGGTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTT TGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGG GATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGA GTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCT TTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTC AAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTG ACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATA CCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGT ACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC ( SEQ ID NO : 312 ; ITCRp clone 93 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSGGGEETQYFGP GTRLLVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSG VCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 313 ; ITCRp clone 93 )

ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGCGCAGGGGGTCAGCGAAAAACTGTTTTTTGGC AGTGGAACCCAGCTCTCTGTCTTGGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGCGGT TTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGA CGGGATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCC GGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTG CCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGAT GTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCC GTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgtgTC ATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTC TGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 314 ; ITCRp clone 96 )

MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSAQGVSEKLFFG SGTQLSVLEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHS GVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF

( SEQ ID NO : 315 ; ITCRP clone 96 )

[0252] In some embodiments, a construct encoding an iTCRp chain comprises an iTCRp V -DJ region comprising a nucleotide sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NOs: 316-385. ACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGA G (SEQ ID NO: 316; iTCRp clone 3)

AGCAGTGCCCCTGGAGGGTCTGAAGCTTTCTTTGGACAAGGCACCAGACTCACAGTTGTAGA G (SEQ ID NO: 317; 1TCR[3 clone 4)

AGCAGTGAACTCGACAGGGAAGGAAACACCATATATTTTGGAGAGGGAAGTTGGCTCACTGT TGTAGAG (SEQ ID NO: 318; 1TCRP clone 7)

AGCAGTGATATGGGACCCGTCTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 319; 1TCRP clone 9)

AGCAGTGAGGAAGAGCGGGGCCCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 320; iTCR[3 clone 13)

AGCAGTGATGGGGTGGGGAGCAATCAGCCCCAGCATTTTGGTGATGGGACTCGACTCTCCAT CCTAGAG (SEQ ID NO: 321; iTCR[3 clone 18)

AGCAGTGACCCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG ( SEQ ID NO: 322; iTCR[3 clone 20)

AGCAGTGAGGCCCCAACAGGAACCGGGGCCAACGTCCTGACTTTCGGGGCCGGCAGCAGGCT GACCGTGCTGGAG (SEQ ID NO: 323; iTCR[3 clone 21)

AGCAGTGATCCTCGACTAGCGGGGGGGGTCGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG (SEQ ID NO: 324; iTCR[3 clone 23)

AGCAGTGAGGGGGCTGGAAACACCATATATTTTGGAGAGGGAAGTTGGCTCACTGTTGTAGA G (SEQ ID NO: 325; iTCR[3 clone 24)

AGCAGTTATGCTACAGGCTACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 326; iTCR[3 clone 26)

AGCAGTGAACGGCAGGGCTCCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 327; iTCR[3 clone 28)

AGCAGTGCTCCGACTAGCGGGAGGGACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCAC CGTGCTAGAG (SEQ ID NO: 328; iTCR[3 clone 29)

AGCAGTGAATGGACTAGCGGGGGGCCCAACACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAG GCTGACCGTACTGGAG (SEQ ID NO: 329; iTCR[3 clone 30)

AGCAGTGAACGGGGGGCTAGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 330; iTCRp clone 32)

AGCAGTGAAGGGCTAGCGGGAGAACCTCTCTTAGGCAATGAGCAGTTCTTCGGGCCAGGGAC ACGGCTCACCGTGCTAGAG (SEQ ID NO: 331; iTCRp clone 33)

AGCAGTGAGGCAGGCGGCCACACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 332; iTCRp clone 34) AGCAGTGAATACCAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGAG (SEQ ID NO: 333; iTCRp clone 36)

AGCACCGACAGGGGATCTTTCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACT GGAG (SEQ ID NO: 334; 1TCR[3 clone 37)

GGAGGAGGGACATCTCAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGA G (SEQ ID NO: 335; 1TCRP clone 38)

AGCAGTCCGACTAGCGGGATGGGGGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGT GCTCGAG (SEQ ID NO: 336; 1TCRP clone 39)

AGCAGTGAGTTCGGGGCCAACGTCCTGACTTTCGGGGCCGGCAGCAGGCTGACCGTGCTGGA G (SEQ ID NO: 337; 1TCRP clone 41)

AGCAGTGTCCGTAGCGGGAGAGGGGACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCAC CGTGCTAGAG (SEQ ID NO: 338; iTCR[3 clone 42)

AGCAGTGTCCAGGAGGAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGA G (SEQ ID NO: 339; iTCR[3 clone 43)

AGCAGTGATAGTAGCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG (SEQ ID NO: 340; iTCR[3 clone 44)

AGCAGTGGTACTACGGGACAGGAATCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG (SEQ ID NO: 341; iTCR[3 clone 46)

AGCAGTGTAAGGGGGAACCACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 342; iTCR[3 clone 47)

AGCAGTGAACTTCAGCGGGAGGGTTCTCCAGATACGCAGTATTTTGGCCCAGGCACCCGGCT GACAGTGCTCGAG (SEQ ID NO: 343; iTCR[3 clone 48)

AGCAGTGTCCGGGACAGGGATGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTT GGAG (SEQ ID NO: 344; iTCR[3 clone 49)

AGCAGTGAGGGTCAGGGAGGTTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCAC AGAG (SEQ ID NO: 345; iTCR[3 clone 50)

AGCAGTGACAGGGATAGCAATCAGCCCCAGCATTTTGGTGATGGGACTCGACTCTCCATCCT AGAG (SEQ ID NO: 346; iTCR[3 clone 51)

AGCAGTGATCGGTCTAGCGGAGCCAAAAACATTCAGTACTTCGGCGCCGGGACCCGGCTCTC AGTGCTGGAG (SEQ ID NO: 347; iTCR[3 clone 52)

AGCAGTGCCACGACTAGCGGGAGGACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGAC AGTGCTGGAG (SEQ ID NO: 348; iTCR[3 clone 53)

AGCAGTGAATTTCGGCAGCGGGAGTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG (SEQ ID NO: 349; iTCRp clone 54) AGCAGTGAAATAGCGGGAGTGGCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 350; iTCRp clone 55)

AGCGTGGGTCCGGTACCCTCCTACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGT GCTAGAG (SEQ ID NO: 351; 1TCRP clone 56)

AGCAGTGAACGGCGCGGGAGACGGGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 352; 1TCRP clone 57)

AGCAGTGGGACAGGGTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG (SEQ ID NO: 353; 1TCRP clone 58)

AGCAGTGACCGTAGCGGGAGCTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 354; iTCR[3 clone 59)

AGCAGTGACAGCACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACTGGAG (SEQ ID NO: 355; iTCR[3 clone 60)

AGCAGTGCTAGCGGGAGCAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCTAGA G (SEQ ID NO: 356; iTCR[3 clone 61)

AGCAGTGACGGGACTAGCGGCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCAC AGAG (SEQ ID NO: 357; iTCR[3 clone 62)

AGCAGTGAATATGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTTGGAG ( SEQ ID NO: 358; iTCR[3 clone 63)

AGCAGTGAGTCCGGCCCCCGCAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 359; iTCR[3 clone 64)

AGCAGTGGCCGACTAGCGGGAGAGGAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCT GGTGCTCGAG (SEQ ID NO: 360; iTCR[3 clone 66)

AGCAGTGAGGGTGGCAGGGTCGATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 361; iTCR[3 clone 67)

AGCAGTGAGGCTAACTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGTAGAG (SEQ ID NO: 362; iTCR[3 clone 68)

AGCAGTCAGGACGGATTGGGATATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGT AGAG (SEQ ID NO: 363; iTCR[3 clone 69)

AGCAGTGGGCGCCTCCACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG (SEQ ID NO: 364; iTCR[3 clone 70)

AGCAGTGAATATAACAGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCT CGAG (SEQ ID NO: 365; iTCR[3 clone 71)

AGCAGTGAACCCGGATTGGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACTGGA G (SEQ ID NO: 366; iTCR[3 clone 72) AGCATCCTGGGAGAGGGGCGGAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 367; iTCRp clone 73)

AGCAGTGCCCCGGGACAGATCTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGT AGAG (SEQ ID NO: 368; 1TCR[3 clone 74)

AGCAGTGACAACCAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGAG (SEQ ID NO: 369; 1TCRP clone 75)

AGCAGTGGGGTGACTAGCGCCTCCTACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCAC CGTGCTAGAG (SEQ ID NO: 370; 1TCRP clone 76)

AGCAGTCCTGAGCCCACCACCCTAGCGGGAGTCCACGAGCAGTACTTCGGGCCGGGCACCAG GCTCACGGTCACAGAG (SEQ ID NO: 371; 1TCRP clone 77)

AGCAGTGGGACACAGAGGGCTGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTT GGAG (SEQ ID NO: 372; 1TCRP clone 78)

AGCAGTGGGACTAGCGGGAGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 373; iTCR[3 clone 79)

AGCAGTGAGGCGGGACAGGGTTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 374; iTCR[3 clone 80)

AGCACCTCTAGCCGCACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACTGGA G (SEQ ID NO: 375; iTCR[3 clone 83)

AGCAGTGAACCGGGGGAGCGGAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 376; iTCR[3 clone 84)

AGCAGTGAAGGTCGGGTTAACTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGT AGAG (SEQ ID NO: 377; iTCR[3 clone 85)

AGCAGTGAATCAGAAGGGGGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 378; iTCRp clone 86)

AGCAGTCCCGGGGGGACTAGCGGGAGGGCACGTCCCTACGAGCAGTACTTCGGGCCGGGCAC CAGGCTCACGGTCACAGAG (SEQ ID NO: 379; iTCRp clone 87)

AGCAGTGGGAGGGAGGGGGACCCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 380; iTCRp clone 88)

AGCAGTGGACTAGCGAACACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACT GGAG (SEQ ID NO: 381; iTCRp clone 89)

AGCAGTGGGACGACAGGGGATACACGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCT GACAGTGCTCGAG (SEQ ID NO: 382; iTCRp clone 90)

AGCAGTGAAGACCGGGACAGGGGTCACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 383; iTCRp clone 91) AGCAGTGAACTAGCGAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCTAGAG ( SEQ ID NO : 384 ; iTCRp clone 92 )

AGCAGTGGAGGAGGGGAGGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGA G ( SEQ ID NO : 385 ; iTCRp clone 93 )

AGCAGTGAATATGCAGGGTGGGGCGGCAATCAGCCCCAGCATTTTGGTGATGGGACTCGACT CTCCATCCTAGAG ( SEQ ID NO : 386 ; iTCRp clone 94 )

AGCAGTGAATTGGACGGGACTAGCGCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG ( SEQ ID NO : 387 ; iTCRp clone 95 )

AGCAGTGCGCAGGGGGTCAGCGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTT GGAG ( SEQ ID NO : 388 ; iTCRp clone 96 )

AGCAGTGAAGTGGCGGGAGCGGACACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGAC CGTACTGGAG ( SEQ ID NO : 389 ; iTCRp clone 97 )

AGCAGCGGCAGGGGGCCAGGGGAAAGTGCAGATACGCAGTATTTTGGCCCAGGCACCCGGCT GACAGTGCTCGAG ( SEQ ID NO : 390 ; iTCRp clone 98 )

B. NK Cells

[0253] The NK cells that are modified to express the TCR/CD3 receptor complex may be obtained from any suitable source, including fresh or frozen. In certain embodiments, NK cells are not NK cells obtained from iPSC differentiation. In certain embodiments, NK cells are derived from human peripheral blood mononuclear cells (PBMC), unstimulated leukapheresis products (PBSC), NK cell lines (e.g., NK-92), human embryonic stem cells (hESCs), induced pluripotent stem cells (iPSCs), bone marrow, or umbilical cord blood by methods well known in the art. Specifically, the NK cells may be isolated from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, NK cell lines, or a mixture thereof. In particular embodiments, the NK cells are isolated from pooled CB. The CB may be pooled from 2, 3, 4, 5, 6, 7, 8, 9, 10, or more units. The NK cells may be autologous or allogeneic with respect to a recipient individual. The isolated NK cells may or may not be haplotype matched for the subject to be administered the cell therapy. NK cells can be detected by specific surface markers, such as CD 16 and CD56 in humans, for example. In some cases, the source of the NK cells is cord blood and the NK cells may be in the cord blood in a heterogeneous mixture of cells and may be depleted of certain cells expressing CD3. In other methods, umbilical CB is used to derive NK cells by the isolation of CD34+ cells.

[0254] The NK cells may be pre-activated with one or more inflammatory cytokines, and they may be expanded or non-expanded. In some cases, the NK cells are pre-activated either prior to modification to express CD3±TCR or following modification to express CD3±TCR complex. In specific embodiments, pre-activation of the NK cells may comprise culturing the isolated NK cells in the presence of one or more cytokines. The NK cells may be stimulated with IL-2, or other cytokines that bind the common gamma-chain (e.g., IL-7, IL-12, IL-15, IL- 18, IL-21, and others). In particular embodiments, the pre-activation cytokines may be selected from the group consisting of IL-12, IL-15, IL-18, and a combination thereof. One or more additional cytokines may be used for the pre-activation step. The pre-activation may be for a short period of time such as 5-72 hours, such as 10-50 hours, particularly 10-20 hours, such as 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours, specifically about 16 hours. The pre-activation culture may comprise IL-12 at a concentration of 0.1-150 ng/mL, such as 0.5-50 ng/mL, particularly 1-20 ng/mL, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ng/mL, specifically about lO ng/mL. The pre-activation culture may comprise IL-18 and/or IL-15 at a concentration of 10-100 ng/mL, such as 40-60 ng/mL, particularly 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 ng/mL, specifically about 50 ng/mL.

[0255] In some cases, the NK cells are expanded either prior to modification to express CD3±TCR complex or following modification to express CD3±TCR complex. Pre-activated NK cells may be expanded in the presence of artificial antigen presenting cells (aAPCs) and/or feeders/fragments or NK activating beads. The pre-activated NK cells may be washed prior to expansion, such as 2, 3, 4, or 5 times, specifically 3 times. The aAPCs may be engineered to express CD137 ligand and/or a membrane-bound cytokine. The membrane-bound cytokine may be membrane-bound IL-21 (mIL-21) or membrane-bound IL- 15 (mIL-15). In particular embodiments, the aAPCs are engineered to express CD137 ligand and mIL-2L The aAPCs may be derived from cancer cells, such as leukemia cells. The aAPCs may not express endogenous HLA class I, II, or CD Id molecules. They may express ICAM-1 (CD54) and LFA- 3 (CD58). In particular, the aAPCs may be K562 cells, such as K562 cells engineered to express CD137 ligand and mIL-21. The aAPCs may be irradiated. In some embodiments, fragments of APC can be used to expand the NK cells. The engineering may be by any method known in the art, such as retroviral transduction. Retroviral transduction may be at least, at most, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days following NK co-culturing with an antigen presenting cell. In some embodiments, retroviral transduction comprises cotransduction of more than one construct. In some embodiments, retroviral transduction occurs after or at about 5 days of co-culturing with an antigen presenting cell. In some embodiments, co-culturing with an antigen presenting cell continues following transduction of an NK cell. The expansion may be for about 2-30 days, such as 3-20 days, particularly 12-16 days, such as 12, 13, 14, 15, 16, 17, 18, or 19 days, specifically about 14 days. The pre-activated NK cells and aAPCs may be present at a ratio of about 3: 1-1 :3, such as 2:1, 1 : 1, 1 :2, specifically about 1 :2. The expansion culture may further comprise cytokines to promote expansion, such as IL- 2. The IL-2 may be present at a concentration of about 10-500 U/mL, such as 100-300 U/mL, particularly about 200 U/mL. The IL-2 may be replenished in the expansion culture, such as every 2-3 days. The aAPCs may be added to the culture at least a second time, such as at about 7 days of expansion.

[0256] In particular embodiments, the NK cells are transfected or transduced with one or more membrane bound cytokines, including IL-21, IL-12, IL-18, IL-23, IL-7, or IL-15, either secreted by NK cells or tethered to the NK cell membrane. In such cases, the membrane bound cytokine may be tethered to the NK cell membrane with a particular transmembrane domain, such as the transmembrane domain of CD8, CD28, CD27, B7H3, IgGl, IgG4, CD4, DAP10, DAP 12, for example.

[0257] Following preparation, the modified NK cells may be immediately infused (including with an effective amount of one or more bispecific or multi-specific antibodies, or the NK cells may be stored, such as by cryopreservation. In some cases, when the NK cells are source from cryopreservation, the NK cells were deactivated pre-cryopreservation using a deactivating agent (e.g., a kinase inhibitor, e.g., Dasatinib, nilotinib, rapamycin, etc.). In certain aspects, the cells may be propagated for days, weeks, or months ex vivo as a bulk population within about 1, 2, 3, 4, or 5 days.

C. Loading of NK Cells

[0258] In particular embodiments, the NK cells are loaded with antibodies prior to use. The NK cells may be loaded in any specific manner, including in culture or immediately before infusion, for example, to produce a complex of NK cells with the antibodies. The conditions are suitable enough to allow for an effective amount of antibody to bind to the surface of the NK cells. In the case of use of monospecific antibodies, the Fc region of the monospecific antibody binds the NK cell while the antigen binding domain of the monospecific antibody is free to bind its target antigen. In certain embodiments of cases of use of multispecific antibodies, one or more antigen binding domains of the antibody can bind the surface of the NK cells, such as through an antigen on the surface of the NK cells, (for example but not limited to, CD3, NKp30, NKp44, NKp46, CD16, CD32, CD64, KIRs, and the like), and the other antigen binding domain is free to bind its target antigen. In certain embodiments of cases of use of multispecific antibodies, one or more antigen binding domains of the antibody can bind one or more target antigens. In certain embodiments of cases of use of multi specific antibodies, the Fc region of the antibody binds the NK cell while the antigen binding domains of the antibody are free to bind target antigens. In certain embodiments, the surface of the NK cell can be recognized by the Fab of an antibody and/or through recognition of the Fc region of the antibody by a cognate receptor. In certain embodiments, an antibody Fc region may be recognized by an endogenous NK cell receptor (e.g., CD16, CD32, CD64 etc.).

[0259] The culture conditions by which the NK cells become loaded may or may not be of a particular type having one or more specific parameters. In particular embodiments, the loading of the NK cells occurs in culture at a specific temperature, such as 37 °C, although in alternative embodiments the temperature is 36 °C or 38 °C, or lower or higher. The duration of the loading step may be for any suitable amount of time, such as in a range of one minute to 24 hours or longer. For example, the range may be in the range of 1 min to 24 hrs, 1 min to 18 hrs, 1 min to 12 hours, 1 min to 6 hrs, 1 min to 1 hr, 30 min to 24 hrs, 30 min to 18 hrs, 30 min to 12 hrs, 30 min to 6 hrs, 30 min to 1 hr, 1-24 hrs, 1-18 hrs, 1-12 hrs, 1-6 hrs, 6-24 hrs, 6-18 hrs, 6-12 hrs, 12-24 hrs, 12-18 hrs, or 18-24 hrs. In some embodiments, the duration of the loading step may be greater than or equal to approximately 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, or 48 hours, or any range derivable therein. In specific embodiments, the cell culture media is basal media or complex media. In some cases, the culture comprises one or more reagents that were utilized during pre-activation and/or expansion steps, while in other cases the culture does not. In specific embodiments, the culture comprises one or more cytokines, including one or more of IL-12, IL-15, IL-2, and IL-18, for example. In some embodiments, the culture comprises APCs of any kind.

[0260] In certain embodiments, antibodies of compositions described herein are subjected in an effective amount to an effective amount of NK cells of the disclosure, thereby producing a complex that is “chimeric antigen receptor-like.” In particular, an antigen binding domain of the antibody binds to the NK cells, such as through the antigen that is a cell surface protein. A plurality of antibodies may be subjected to a plurality of NK cells such that there are multiple complexes of cell/antibody. The antibodies may be of any type, including monospecific, bi specific, or multi specific, and in specific cases the antibody engages both the NK cell and a target antigen through an antigen binding domain of the antibody (such as with engagers in the art that are fusion proteins consisting of two single-chain variable fragments (scFvs) of different antibodies). In examples wherein the antibody is monospecific, an antigen binding domain of the antibody binds a target antigen, such as a cancer antigen, and another part of the antibody binds the NK cells, such as an Fc region of the antibody. In cases wherein the antibody is multispecific, one or more antigen binding domains of the antibody may bind the NK cell (such as through an NK cell surface antigen, either naturally occurring, or transgenic, e.g., CD3) and one or more antigen binding domains of the antibody binds one or more target antigens. In certain embodiments of cases of use of multispecific antibodies, one or more antigen binding domains of the antibody and/or the Fc region of the antibody may bind the NK cell. The multispecific antibody may be bispecific, trispecific, or tetraspecific, for example. In cases wherein the antibody is trispecific or tetraspecific, the additional antigen binding domains may bind other cells, such as stem cells.

[0261] In particular embodiments, the antibodies may bind any NK cell surface antigen (that may or may not be receptors) on NK cells, such as CD 16 (including CD 16a or CD 16b), CD32, CD56, CD64, a c-type lectin such as NKG2D, NKG2C, a costimulatory molecule such as CS1, DNAM, 2B4, CD2, an NCR, NKp30, NKp44, NKp46, or KIR, and redirect the NK cells to a target, thus increasing the response and specificity against different tumors. In certain embodiments, the antibodies may bind to a transgenic NK cell surface antigen, such as CD3.

[0262] In some embodiments, the antibodies may bind any suitable antigen (e.g., antigens described herein, such as those that are described as targets of CARs and/or TCRs). In particular embodiments, an antibody targets CD 19. In particular embodiments, an antibody targets CD20. In particular embodiments, an antibody targets CD123. In particular embodiments, an antibody targets EGFR. In particular embodiments, an antibody targets EGFR2.

[0263] In certain embodiments, generation of loaded NK cells may be by any suitable means, such that the conditions are sufficient for the appropriate region of the antibody to bind the appropriate surface region of the NK cell. Any particular medium may be utilized, in certain instances. In specific cases, Plasma-Lyte A and/or human serum albumin are utilized, wherein in other cases they are not. Once the complexes are formed in culture, they may or may not be washed prior to administration to the subject, such as through infusion. In alternative embodiments, the NK cells and the antibodies are administered separately, and the complexes form in vivo.

D. Pre-Activation

[0264] In some embodiments, the NK cells are pre-activated prior to administration to a recipient individual. The pre-activation step may or may not occur before any expansion step. In specific embodiments, the NK cells are pre-activated with one or more cytokines, and in specific embodiments, the NK cells are pre-activated with one or more of IL-12, IL-15, IL-2, and IL- 18 and including two, three, or more. In cases wherein less than all three of IL- 12, IL- 15, IL-2, and IL-18 are utilized, it may be that IL-12 and IL-15 but not IL-18; or IL-12 and IL- 18 but not IL-15; or IL-15 and IL-18 but not IL-12. IL-2 may or may not be substituted for IL- 15.

[0265] In particular embodiments, the pre-activation cytokines may be IL-12, IL-15, and IL-18. One or more additional cytokines may be used for the pre-activation step. The preactivation may be for a short period of time such as 5-72 hours, such as 10-50 hours, particularly 10-20 hours, such as 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours, and specifically about 16 hours in some cases. The pre-activation culture may comprise IL-18 and/or IL-15 at a concentration of 10-100 ng/mL, such as 40-60 ng/mL, particular 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 ng/mL, specifically about 50 ng/mL. In some cases, the pre-activation culture comprises IL-12 at a concentration of 0.1-150 ng/mL, including at a concentration of 1-20 ng/mL, such as a concentration of 10 ng/mL. In alternative embodiments the NK cells may be stimulated with IL-2, or other cytokines that bind the common gamma-chain (e.g., IL-7, IL-21, and others), and this may be in addition to IL-12, IL-15, and IL-18 or as an alternative to one or more of them. In such cases, the pre-activation culture may comprise IL- 12 at a concentration of 0.1-150 ng/mL, such as 0.5-50 ng/mL, particularly 1-20 ng/mL, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ng/mL, specifically about 10 ng/mL.

E. Expansion

[0266] In particular embodiments, NK cells are expanded to increase their quantity prior to administration to an individual in need thereof. The expanded cells may or may not be derived from pre-activated NK cells such that a pre-activation step may occur before an expansion step. The NK cell expansion step may be of any suitable such that the NK cell population is expanded, but in specific cases the expansion step utilizes particular one or more reagents, such as in culture, to enhance their expansion. In certain cases the NK cells may not be expanded. IL-2 or IL- 15 or IL- 18 or any combination of the cytokines may be added to the expansion culture before or during expansion. The NK cells can be expanded ex vivo in flasks or in one of several different bioreactor configurations with continuous perfusion of media/additives, in specific embodiments.

[0267] In specific cases, the NK cells (whether pre-activated or not) may be washed (e.g., with PBS or Plasma Lyte or human serum albumin or culture media or combinations thereof) prior to and/or after expansion, such as 1, 2, 3, 4, or 5 times. In some embodiments, cells are washed specifically 3 times. In particular embodiments, the NK cells are expanded in the presence of artificial antigen presenting cells (aAPCs). In particular embodiments, the NK cells are expanded in the presence of fragments of aAPCs. The aAPCs may be engineered to express CD137 ligand and/or a membrane-bound cytokine. The membrane-bound cytokine may be membrane-bound IL-21 (mIL-21) or membrane-bound IL- 15 (mIL-15). In particular embodiments, the aAPCs are engineered to express CD137 ligand and mIL-21. The aAPCs may be derived from cancer cells, such as leukemia cells. The aAPCs may not express endogenous HLA class I, II, or CD Id molecules. They may express ICAM-1 (CD54) and LFA- 3 (CD58) or CD48. In particular, the aAPCs may be K562 cells, such as K562 cells engineered to express CD137 ligand and mIL-21. The engineering may be by any method known in the art, such as retroviral transduction, although any viral or non-viral vector may be utilized. The aAPCs may or may not be irradiated. The expansion may be for a particular duration in time, such as for about 2-30 days, such as 3-20 days, particularly 12-16 days, such as 12, 13, 14, 15, 16, 17, 18, or 19 days, specifically about 14 days. The pre-activated NK cells and aAPCs may be present at a ratio ofabout 3: 1-1 :3, such as 2: 1, 1 : 1, 1 :2, specifically about 1 :2. The expansion culture may further comprise one or more cytokines to promote expansion, such as IL-2. The IL-2 may be present at a concentration of about 10-500 U/mL, such as 100-300 U/mL, particularly about 200 U/mL. The IL-2 may be replenished in the expansion culture, including at a certain frequency, such as every 2-3 days. The aAPCs may be added to the culture at least a second time, such as at about 7 days of expansion. Any cytokine(s) used in the pre-activation and/or expansion steps may be recombinant human cytokines.

[0268] In some embodiments, following expansion, the NK cells may be immediately utilized in any manner, such as complexed with one or more antibodies, or they may be stored, such as by cryopreservation. In certain aspects, the cells may be propagated for days, weeks, or months ex vivo as a bulk population within about 1, 2, 3, 4, or 5 days.

[0269] Activated and/or expanded NK cells can secrete type I cytokines, such as interferon- y, tumor necrosis factor-a and granulocyte-macrophage colony-stimulating factor (GM-CSF), which activate both innate and adaptive immune cells as well as other cytokines and chemokines. The measurement of these cytokines can be used to determine the activation status of NK cells. In addition, other methods known in the art for determination of NK cell activation may be used for characterization of the NK cells of the present disclosure.

[0270] Thus, with respect to particular pre-activation and expansion aspects of the disclosure, in specific embodiments the NK cells pre-activated with any combination of IL-12, IL15, and/or IL-18 followed by expansion with aAPCs, such as K562 cells expressing mIL-21 and CD 137 ligand, provide a highly potent cellular product. Thus, methods are provided using the present NK cells for the treatment of various diseases, such as immunotherapy of patients

- I l l - with cancer. In an exemplary method, the isolated NK cells may be subjected to a brief period, such as about 16 hours, of pre-activation with a combination of cytokines, such as interleukin- 12 (IL-12), IL-15, and/or IL-18, followed by expansion using artificial antigen presenting cells (aAPCs), such as K562 feeder cells expressing membrane-bound IL-21 and CD137 ligand, and/or exogenous IL-2. IL-2 or IL- 15 or IL- 18 or any combination of the cytokines may be added to the expansion culture at least a second time.

F. Cryopreservation

[0271] In particular cases, NK cells and/or antibodies of the disclosure are preserved in a cry opreservation medium composition comprising at least one cryoprotectant, a serum (human or animal serum) or a non-serum alternative to serum (not human serum or animal serum), and at least one cytokine and/or at least one growth factor. In some cases, the cryoprotectant is dimethyl sulfoxide (DMSO), glycerin, glycerol, hydroxy ethyl starch, or a combination thereof. The non-serum alternative may be of any kind, including at least platelet lysate and/or a blood product lysate (for example, human serum albumin). In embodiments of the composition wherein one or more (including two or more) cytokines are utilized, the cytokine may be a natural or a recombinant or a synthetic protein. At least one of the cytokines may be an Food and Drug Administration (FDA)-approved cytokine. Examples of cytokines and growth factors include at least IL-1, IL-2, IL-3, IL-4, IL-6, IL-7, IL-9, IL-10, IL-12, IL-13, IL-15, IL- 17, IL-18, IL-21, IL-22, interferon, tumor necrosis factor, stem cell factor, FLT3-ligand, APRIL, thrombopoietin, erythropoietin, or a combination thereof. For serum embodiments, the serum may be an animal-derived serum, such as human serum (including human AB serum) or bovine serum. DMSO and other cryoprotectants, when utilized may comprise 4-10%, 4- 6%, 4-8%, 5-10%, 5-8%, 6-10%, 6-8%, 8-10%, and so forth, of the composition. For embodiments wherein serum is employed, the serum may comprise 5-99%, 5-95%, 5-90%, 5- 85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5- 25%, 5-20%, 5-15%, 5-10%, 10-99%, 10-95%, 10-90%, 10-85%, 10-80%, 10-75%, 10-70%, 10-65%, 10-60%, 10-55%, 10-50%, 10-45%, 10-40%, 10-35%, 10-30%, 10-25%, 10-20%, 10- 15%, 20-99%, 20-95%, 20-90%. 20-85%, 20-80%, 20-75%, 20-70%, 20-65%, 20-60%, 20-

55%, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 20-25%, 30-99%, 30-95%, 30-90%, 30-

85%, 30-80%, 30-75%, 30-70%, 30-65%, 30-60%, 30-55%, 30-50%, 30-45%, 30-40%, 30-

35%, 40-99%, 40-95%, 40-90%, 40-85%, 40-80%, 40-75%, 40-70%, 40-65%, 40-60%, 40-

55%, 40-50%, 40-45%, 50-99%, 50-95%, 50-90%, 50-85%, 50-80%, 50-75%, 50-70%, 50-

65%, 50-60%, 50-55%, 60-99%, 60-95%, 60-90%, 60-85%, 60-80%, 60-75%, 60-70%, 60- 65%, 70-99%, 70-95%, 70-90%, 70-85%, 70-80%, 70-75%, 80-99%, 80-95%, 80-90%, 80- 85%, 90-99%, 90-95%, or 95-99% of the composition. The composition may comprise at least or no more than 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of serum. In specific embodiments, the composition comprises platelet lysate that may be at any concentration in the composition, but in certain embodiments the platelet lysate comprises 5-99%, 5-95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5- 60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-99%, 10-95%, 10-90%, 10-85%, 10-80%, 10-75%, 10-70%, 10-65%, 10-60%, 10-55%, 10-50%, 10- 45%, 10-40%, 10-35%, 10-30%, 10-25%, 10-20%, 10-15%, 20-99%, 20-95%, 20-90%. 20-

85%, 20-80%, 20-75%, 20-70%, 20-65%, 20-60%, 20-55%, 20-50%, 20-45%, 20-40%, 20-

35%, 20-30%, 20-25%, 30-99%, 30-95%, 30-90%, 30-85%, 30-80%, 30-75%, 30-70%, 30-

65%, 30-60%, 30-55%, 30-50%, 30-45%, 30-40%, 30-35%, 40-99%, 40-95%, 40-90%, 40-

85%, 40-80%, 40-75%, 40-70%, 40-65%, 40-60%, 40-55%, 40-50%, 40-45%, 50-99%, 50-

95%, 50-90%, 50-85%, 50-80%, 50-75%, 50-70%, 50-65%, 50-60%, 50-55%, 60-99%, 60-

95%, 60-90%, 60-85%, 60-80%, 60-75%, 60-70%, 60-65%, 70-99%, 70-95%, 70-90%, 70-

85%, 70-80%, 70-75%, 80-99%, 80-95%, 80-90%, 80-85%, 90-99%, 90-95%, or 95-99% of the composition. The composition may comprise at least or no more than 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of platelet lysate.

[0272] The composition may have certain concentrations of components, including cytokines and/or growth factors. In specific cases, any cytokine, including IL-2, IL-21, and/or IL- 15, for example, are present in the composition in a particular concentration. The IL-2 may be present at a concentration of 1-5000, 1-1000, 1-500, 1-100, 100-5000, 100-500, 500-5000, 500-1000, or 1000-5000 U/mL, for example. In a specific case, the IL-2 is present at a concentration in the composition of at least or no more than 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 U/mL. In specific embodiments, IL-21 is present in the composition at a concentration of 10-3000, 10-2000, 10-1000, 10-500, 10-100, 100-3000, 100-2000, 100-1000, 500-3000, 500-2000, 500-1000, 1000-3000, 1000-2000, or 2000-3000 ng/mL. The IL-21 may be in a concentration in the composition of at least or nor more than 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 750, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, or 3000 ng/mL. IL-15 may be present in the composition at a concentration of 1-2000, 1-1000, 1-500, 1-100, 100-2000, 100-1000, 100-500, 500-2000, 500-1000, or 1000- 2000 ng/mL. IL- 15 may be present in the composition at a concentration of at least or no more than 10, 50, 100, 500, 1000, 1500, or 2000 ng/mL. [0273] Compositions as encompassed herein that comprise at least one cryoprotectant, a serum or a non-serum alternative to serum, and at least one cytokine and/or at least one growth factor may further comprise a plurality of immune cells and/or stem cells, each of any kind. In specific embodiments, the cells are NK cells, T cells, B cells, NKT cells derived from mature bone marrow or peripheral blood cells, cell lines such as tumor cell lines (e.g., NK92 or other NK lines), hematopoietic stem cells, induced pluripotent stem cells, MSCs (a population of cells alternatively called “mesenchymal stem cells” and “mesenchymal stromal cells” in the literature), or a mixture thereof, which can be derived from bone marrow, peripheral blood, skin, adipose tissue, or a combination thereof. In embodiments wherein NK cells are utilized, the NK cells may or may not be expanded NK cells. Embodiments of the disclosure also encompass pharmaceutical compositions that comprise any composition of the disclosure and a suitable pharmaceutically acceptable carrier.

[0274] In certain embodiments, cells and/or antibodies are treated with one or more deactivating agents (e.g., a kinase inhibitor, e.g., Dasatinib, Nilotinib, Rapamycin, etc.) precryopreservation.

[0275] In some embodiments, technologies described herein comprise deactivating a NK cell, comprising treating an NK cell with an effective amount of one or more deactivating agents under conditions to produce a deactivated NK cell. In some embodiments, a deactivating agent is a kinase inhibitor. In some embodiments, a deactivating agent is a mechanistic target of rapamycin (mTOR) inhibitor. In some embodiments, the mTOR inhibitor is rapamycin, everolimus, and/or temsirolimus. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the deactivating agent is a tyrosine kinase (TK) inhibitor. In some embodiments, the TK inhibitor is Lorlatinib, Brigatinib, Ceritinib, Alectinib, Crizotinib, Bosutinib, Ponatinib, Nilotinib, Dasatinib, Imatinib, Zanubrutinib, Acalabrutinib, Ibrutinib, Capmatinib, Pexidartinib, Dacomitinib, Osimertinib, Erlotinib, Gefitinib, Lapatinib, Afatinib, Pemigatinib, Erdafitinib, Nintedanib, Gilteritinib, Midostaurin, Tucatinib, Neratinib, Baricitinib, Ruxolitinib, Fedratinib, Tofacitinib, Ripretinib, Selumetinib, Binimetinib, Cobimetinib, Trametinib, Upadacitinib, Avapritinib, Selpercatinib, Cabozantinib, Fostamatinib, Larotrectinib, Entrectinib, Axitinib, Regorafenib, Pazopanib, Sorafenib, Lenvatinib, Vandetanib, and/or Sunitinib. In some embodiments, the TK inhibitor is a BCR- Abl inhibitor. In some embodiments, the TK inhibitor is Bosutinib, Ponatinib, Nilotinib, Dasatinib, and/or Imatinib. In some embodiments, the TK inhibitor is Dasatinib and/or Nilotinib. In some embodiments, the TK inhibitor is Dasatinib. [0276] In some embodiments, treatment with a deactivating agent is at any point during culturing of the NK cell. In some embodiments, the treatment is for about 24 to about 96 hours, about 36 to about 84 hours, or about 48 to about 72 hours. In some embodiments, the treatment is for about 24 hours, about 48 hours, or about 72 hours. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 1 to about 1000 nM. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 5 to about 500 nM. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 20 to about 200 nM. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 30 to about 100 nM. In some embodiments, the deactivated NK cell has an increased expression of one or more of C-kit, CCR-5, CD62L and/or CXCR4, and/or decreased expression of one or more of NKG2D, DNAM, OX-40, TRAIL, HLA-DR, CD2, CD25, ICOS, and/or CD95 relative to an activated NK cell.

In some embodiments, technologies described herein comprise methods of maintaining the viability of a population of cells over at least 50% percent following cryopreservation of the population, comprising the step of subjecting the population to an effective amount of one or more deactivating agents (e.g., a tyrosine kinase inhibitor) to deactivate the cells prior to cryopreservation, cryopreserving the cells, and thawing the population, wherein upon thawing the viability of the population is over at least 50%. In some cases, upon thawing of the cells the viability of the population of cells is over at least 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% following cryopreservation of the population.

III. Heterologous Proteins and Mutations

[0277] In specific embodiments, the NK cells are modified not only to express one or more components of the TCR/CD3 complex, but they are also modified to express one or more other heterologous proteins. The heterologous proteins may facilitate activity of the NK cells in any manner, including at least their activation, persistence, expansion, homing, and/or cytotoxicity.

A. Bispecific or Multi-specific Antibodies

[0278] In some embodiments, the NK cells are modified to express one or more bispecific or multi-specific antibodies, although in other cases the NK cells do not express the antibodies but the antibodies are utilized in conjunction with the NK cells.

[0279] In certain cases wherein the NK cells are modified to express the antibodies and/or are utilized in conjunction with the NK cells, the antibodies may be any antibody or antibodylike structure known in the art, including antibody fragments, single-domain antibodies, scFv, bispecific antibodies, bispecific diabodies, trispecific antibodies, scFv-Fc and other antibody constructs and engagers. In particular, the antibodies may be engagers that bridge a particular immune effector cell with a particular target cell for destruction of the target cell. The present disclosure allows the modified NK cells to be used with standard T-cell engagers (BiTEs) because they have been modified to express CD3 that in many cases is the T cell antigen to which the BiTE engager binds. In such cases, the BiTE used in the invention may also target a cancer or viral antigen that may be tailored to the medical condition of an intended recipient individual. For example, the BiTE may be tailored to bind a cancer antigen that is characteristic of the cancer cells of a cancer of the individual. The anti-CD3 antibody of the BiTE may target the CD3y chain, CD36 chain, CD3s chain, or CD3^ chain.

[0280] In some cases, in addition to expressing the CD3 complex (with or without TCR) that allows the NK cells to be utilized as a therapy with BiTEs, the NK cells may be modified to express (or not to express but instead used in conjunction with) one or more bispecific NK engagers (BiKEs). The BiKE comprises an antibody that binds a surface protein on the NK cell, including a naturally expressed surface protein on NK cells (for example but not limited to, NKp30, NKp44, NKp46, CD 16, CD32, CD64, KIRs, and the like), and also comprises an antibody that binds a desired target antigen. The BiKE may target the NK cells through an antibody an NK surface protein such as CD16, CS1, CD32, CD64, CD56, NKG2D, NKG2C, DNAM, 2B4, CD2, an NCR, NKp30, NKp44, NKp46, or KIR, for example. In such cases, the BiKE used in the invention may also target a cancer or viral antigen that may be tailored to the medical condition of an intended recipient individual. For example, the BiKE may be tailored to bind a cancer antigen that is characteristic of the cancer cells of a cancer of the individual.

[0281] In certain embodiments, an antibody is Blinatumomab. In certain embodiments, an antibody is Tebentafusp. In certain embodiments, an antibody is Mosunetuzumab. In certain embodiments, an antibody is Teclistamab. In certain embodiments, an antibody is Glofitamab. In certain embodiments, an antibody is Epcoritamab. In some embodiments, an antibody is Flotetuzumab. In some embodiments, an antibody is APV0436. In some embodiments, an antibody is TNB383B. In certain embodiments of cases of use of multispecific antibodies, one or more antigen binding domains of the antibody can bind one or more target antigens.

[0282] In embodiments wherein an NK cell expresses the CD3 complex (with or without TCR and/or iTCR) and one or more BiKEs, one or more vectors may be utilized to transfect or transduce the cells with the CD3 complex components (with or without TCR and/or iTCR) and one or more BiKEs. In some cases, one or more of the CD3 complex components (with or without TCR and/or iTCR) and the BiKE may or may not be on the same multicistronic vector.

B. Engineered Receptors

[0283] In specific embodiments, the NK cells are engineered to express one or more engineered receptors. In some cases, the engineered receptors are engineered antigen receptors that target a cancer or viral antigen of any kind. The receptor may be tailored to target a desired antigen based on a medical condition of an intended recipient individual.

[0284] In some embodiments, the engineered antigen receptor is a chimeric antigen receptor (CAR). The NK cells may be modified to encode at least one CAR, and the CAR may be first generation, second generation, or third or a subsequent generation, for example. The CAR may or may not be bispecific for two or more different antigens. The CAR may comprise one or more costimulatory domains. Each costimulatory domain may comprise the costimulatory domain of any one or more of, for example, members of the TNFR superfamily, CD28, CD137 (4-1BB), CD134 (0X40), DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA- 1 (CDl la/CD18), Lek, TNFR-I, TNFR-II, Fas, CD30, CD27, NKG2D, 2B4M, CD40 or combinations thereof, for example. In specific embodiments, the CAR comprises CD3zeta. In certain embodiments, the CAR lacks one or more specific costimulatory domains; for example, the CAR may lack 4-1BB and/or lack CD28.

[0285] In particular embodiments, the CAR polypeptide in the cells comprises an extracellular spacer domain that links the antigen binding domain and the transmembrane domain, and this may be referred to as a hinge. Extracellular spacer domains may include, but are not limited to, Fc fragments of antibodies or fragments or derivatives thereof, hinge regions of antibodies or fragments or derivatives thereof, CH2 regions of antibodies, CH3 regions antibodies, artificial spacer sequences or combinations thereof. Examples of extracellular spacer domains include but are not limited to CD8-alpha hinge, CD28, artificial spacers made of polypeptides such as Gly3, or CHI, CH3 domains of IgGs (such as human IgGl or IgG4). In specific cases, the extracellular spacer domain may comprise (i) a hinge, CH2 and CH3 regions of IgG4, (ii) a hinge region of IgG4, (iii) a hinge and CH2 of IgG4, (iv) a hinge region of CD8-alpha or CD4, (v) a hinge, CH2 and CH3 regions of IgGl, (vi) a hinge region of IgGl or (vii) a hinge and CH2 of IgGl, (viii) a hinge region of CD28, or a combination thereof. In specific embodiments, the hinge is from IgGl and in certain aspects the CAR polypeptide comprises a particular IgGl hinge amino acid sequence or is encoded by a particular IgGl hinge nucleic acid sequence. [0286] The transmembrane domain in the CAR may be derived either from a natural or from a synthetic source. Where the source is natural, the domain in some aspects is derived from any membrane-bound or transmembrane protein. Transmembrane regions include those derived from (z.e., comprise at least the transmembrane region(s) of) the alpha, beta or zeta chain of the T- cell receptor, CD28, CD3 zeta, CD3 epsilon, CD3 gamma, CD3 delta, CD45, CD4, CD5, CD8, CD9, CD 16, CD22, CD33, CD37, CD64, CD80, CD86, CD 134, CD137, CD154, ICOS/CD278, GITR/CD357, NKG2D, and DAP molecules, such as DAP10 or DAP12. Alternatively the transmembrane domain in some embodiments is synthetic. In some aspects, the synthetic transmembrane domain comprises predominantly hydrophobic residues such as leucine and valine. In some aspects, a triplet of phenylalanine, tryptophan and valine may be found at each end of a synthetic transmembrane domain.

[0287] In some embodiments, the engineered receptors utilize one or more homing receptors (that can home to a target not necessarily because of a signal release, such as in the event that they utilize adhesion molecules) and/or one or more chemokine receptors. Examples of chemokine receptors include CXC chemokine receptors, CC chemokine receptors, CX3C chemokine receptors and XC chemokine receptors. In specific cases, the chemokine receptor is a receptor for CCR2, CCR3, CCR5, CCR8, CCR7, CXCR3, L-selectin (CD62L) CXCR1, CXCR2, or CX3CR1.

C. Cytokines

[0288] In some embodiments, the cells expressing the NK cells are engineered to express one or more heterologous cytokines and/or are engineered to upregulate normal expression of one or more heterologous cytokines. The cells may or may not be transduced or transfected for one or more cytokines on the same vector as other genes. In certain embodiments, NK cells may be modified to express one or more cytokines, cytokine receptors, chemokines, chemokine receptors, and/or suicide genes.

[0289] One or more cytokines may be co-expressed from a vector, including as a separate polypeptide from any component of the TCR/CD3 complex. Interleukin- 15 (IL- 15), for example, is tissue restricted and only under pathologic conditions is it observed at any level in the serum, or systemically. IL- 15 possesses several attributes that are desirable for adoptive therapy. IL-15 is a homeostatic cytokine that induces development and cell proliferation of natural killer cells, promotes the eradication of established tumors via alleviating functional suppression of tumor-resident cells, and inhibits activation-induced cell death (AICD). In addition to IL- 15, other cytokines are envisioned. These include, but are not limited to, cytokines, chemokines, and other molecules that contribute to the activation and proliferation of cells used for human application. NK cells expressing IL- 15 are capable of continued supportive cytokine signaling, which is useful for their survival post-infusion.

[0290] In some embodiments, the cells express one or more exogenously provided engineered receptors, wherein the engineered receptor comprises a chemokine receptor and/or a cytokine receptor. In some embodiments, a cytokine receptor is an IL- 15 receptor. In some embodiments, a cytokine receptor is a non-naturally occurring variant of a cytokine receptor. In some embodiments, a cytokine receptor is an IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, or GMCSF receptor, or a combination thereof.

[0291] In specific embodiments, the cells express one or more exogenously provided cytokines. As one example, the cytokine is IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, GMCSF, or a combination thereof. The cytokine may be exogenously provided to the NK cells because it is expressed from an expression vector within the cell. In an alternative case, an endogenous cytokine in the cell is upregulated upon manipulation of regulation of expression of the endogenous cytokine, such as genetic recombination at the promoter site(s) of the cytokine. In cases wherein the cytokine is provided on an expression construct to the cell, the cytokine may be encoded from the same vector as one or more components of the CD3 complex with or without the TCR complex.

[0292] In some embodiments, a specific sequence of IL-15 is utilized, such as those that follow (underlining refers to signal peptide sequence): ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCT GAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCG GACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGAC CTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTG CAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCG ACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGC AACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAA AGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 49 )

MRISKPHLRS IS IQCYLCLLLNSHFLTEAG I HVF I LGC FS AGL PKTE ANWVNVI S DLKK I E D LIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDAS IHDTVENLI ILANNSLSS NGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS ( SEQ ID NO : 48 )

D. Antigens

[0293] The modified NK cells of the disclosure are utilized with bispecific or multi-specific antibodies that target one or more particular antigens. In addition, the NK cells may be modified with engineered antigen receptors that target one or more particular antigens. In cases wherein the NK cells are modified with one or more engineered antigen receptors, the antigen targeted by the bispecific or multi-specific antibody, and the antigen targeted by the one or more engineered antigen receptors may or may not be the same antigen. In some cases, the antigen targeted by the bispecific or multi-specific antibody, and the antigen targeted by the one or more engineered antigen receptors are different antigens but are associated with the same type of cancer.

[0294] Among the antigens targeted by the antibodies and/or engineered antigen receptors are those expressed in the context of a disease, condition, or cell type to be targeted via the adoptive cell therapy. Among the diseases and conditions are proliferative, neoplastic, and malignant diseases and disorders, including cancers and tumors, including hematologic cancers, cancers of the immune system, such as lymphomas, leukemias, and/or myelomas, such as B, T, and myeloid leukemias, lymphomas, and multiple myelomas. In some embodiments, the antigen is selectively expressed or overexpressed on cells of the disease or condition, e.g., the tumor or pathogenic cells, as compared to normal or non-targeted cells or tissues. In other embodiments, the antigen is expressed on normal cells and/or is expressed on the engineered cells.

[0295] Any suitable antigen may be targeted in the present method. The antigen may be associated with certain cancer cells but not associated with non-cancerous cells, in some cases. Exemplary antigens include, but are not limited to, antigenic molecules from infectious agents, auto-/self-antigens, tumor-/cancer-associated antigens, and tumor neoantigens (Linnemann et al., 2015). In particular aspects, the antigens include NY-ESO, CD 19, EBNA, CD 123, HER2, CA-125, TRAIL/DR4, CD20, CD22, CD70, CD38, CD123, CLL1, carcinoembryonic antigen, alphafetoprotein, CD56, AKT, Her3, epithelial tumor antigen, CD319 (CS1), ROR1, folate binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, CD5, CD23, CD30, HERV-K, IL-1 IRalpha, kappa chain, lambda chain, CSPG4, CD33, CD47, CLL-1, U5snRNP200, CD200, BAFF-R, BCMA, CD99, p53, mutated p53, Ras, mutated ras, c-Myc, cytoplasmic serine/threonine kinases (e.g., A-Raf, B-Raf, and C-Raf, cyclin-dependent kinases), MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A10, MAGE- A12, MART-1, melanoma-associated antigen, BAGE, DAM-6, -10, GAGE-1, -2, -8, GAGE- 3, -4, -5, -6, -7B, NA88-A, MC1R, mda-7, gp75, GplOO, PSA, PSM, Tyrosinase, tyrosinase- related protein, TRP-1, TRP-2, ART-4, CAMEL, CEA, Cyp-B, hTERT, hTRT, iCE, MUC1, MUC2, Phosphoinositide 3-kinases (PI3Ks), TRK receptors, PRAME, P15, RU1, RU2, SART- 1, SART-3, Wilms' tumor antigen (WT1), AFP, -catenin/m, Caspase-8/m, CDK-4/m, ELF2M, GnT-V, G250, HAGE, HSP70-2M, HST-2, KIAA0205, MUM-1, MUM-2, MUM-3, Myosin/m, RAGE, SART-2, TRP-2/INT2, 707-AP, Annexin II, CDC27/m, TPI/mbcr-abl, BCR-ABL, interferon regulatory factor 4 (IRF4), ETV6/AML, LDLR/FUT, Pml/RAR, Tumor- associated calcium signal transducer 1 (TACSTD1) TACSTD2, receptor tyrosine kinases (e.g., Epidermal Growth Factor receptor (EGFR) (in particular, EGFRvIII), platelet derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor (VEGFR)), VEGFR2, cytoplasmic tyrosine kinases (e.g., src-family, syk-ZAP70 family), integrin-linked kinase (ILK), signal transducers and activators of transcription STAT3, STATS, and STATE, hypoxia inducible factors (e.g., HIF-1 and HIF-2), Nuclear Factor-Kappa B (NF-B), Notch receptors (e.g., Notchl-4), NY ESO 1, c-Met, mammalian targets of rapamycin (mTOR), WNT, extracellular signal-regulated kinases (ERKs), and their regulatory subunits, PMSA, PR-3, MDM2, Mesothelin, renal cell carcinoma-5T4, SM22-alpha, carbonic anhydrases I (CAI) and IX (CAIX) (also known as G250), STEAD, TEL/AML1, GD2, proteinase3, hTERT, sarcoma translocation breakpoints, EphA2, ML-IAP, EpCAM, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, GD3, fucosyl GM1, mesothelian, PSCA, sLe, PLAC1, GM3, BORIS, Tn, GLoboH, NY-BR-1, RGsS, SAGE, SART3, STn, PAX5, OY-TES1, sperm protein 17, LCK, HMWMAA, AKAP- 4, SSX2, XAGE 1, B7H3, legumain, TIE2, Page4, MAD-CT-1, FAP, MAD-CT-2, fos related antigen 1, CBX2, CLDN6, SPANX, TPTE, ACTL8, ANKRD30A, CDKN2A, MAD2L1, CTAG1B, SUNCI, and LRRN1. Examples of sequences for antigens are known in the art, for example, in the GENBANK® database: CD19 (Accession No. NG_007275.1), EBNA (Accession No. NG_002392.2), WT1 (Accession No. NG_009272.1), CD123 (Accession No. NC_000023.11), NY-ESO (Accession No. NC_000023.11), EGFRvIII (Accession No. NG_007726.3), MUC1 (Accession No. NG_029383.1), HER2 (Accession No. NG_007503.1), CA-125 (Accession No. NG_055257.1), WT1 (Accession No. NG_009272.1), Mage-A3 (Accession No. NG_013244.1), Mage-A4 (Accession No. NG_013245.1), Mage-AlO (Accession No. NC_000023.11), TRAIL/DR4 (Accession No. NC_000003.12), and/or CEA (Accession No. NC_000019.10).

[0296] Tumor-associated antigens may be derived from prostate, breast, colorectal, lung, pancreatic, renal, mesothelioma, ovarian, liver, brain, bone, stomach, spleen, testicular, cervical, anal, gall bladder, thyroid, or melanoma cancers, as examples. Exemplary tumor- associated antigens or tumor cell-derived antigens include MAGE 1, 3, and MAGE 4 (or other MAGE antigens such as those disclosed in International Patent Publication No. WO 99/40188); PRAME; BAGE; RAGE, Lage (also known as NY ESO 1); SAGE; and HAGE or GAGE. These non-limiting examples of tumor antigens are expressed in a wide range of tumor types such as melanoma, lung carcinoma, sarcoma, and bladder carcinoma. See, e.g., U.S. Patent No. 6,544,518. Prostate cancer tumor-associated antigens include, for example, prostate specific membrane antigen (PSMA), prostate-specific antigen (PSA), prostatic acid phosphates, NKX3.1, and six-transmembrane epithelial antigen of the prostate (STEAP).

[0297] Other tumor associated antigens include Plu-1, HASH-1, HasH-2, Cripto and Criptin. Additionally, a tumor antigen may be a self-peptide hormone, such as whole length gonadotrophin hormone releasing hormone (GnRH), a short 10 amino acid long peptide, useful in the treatment of many cancers.

[0298] Antigens may include epitopic regions or epitopic peptides derived from genes mutated in tumor cells or from genes transcribed at different levels in tumor cells compared to normal cells, such as telomerase enzyme, survivin, mesothelin, mutated ras, bcr/abl rearrangement, Her2/neu, mutated or wild-type p53, cytochrome P450 1B1, and abnormally expressed intron sequences such as N-acetylglucosaminyltransferase-V; clonal rearrangements of immunoglobulin genes generating unique idiotypes in myeloma and B-cell lymphomas; tumor antigens that include epitopic regions or epitopic peptides derived from oncoviral processes, such as human papilloma virus proteins E6 and E7; Epstein bar virus protein LMP2; nonmutated oncofetal proteins with a tumor-selective expression, such as carcinoembryonic antigen and alpha-fetoprotein.

E. Suicide Gene

[0299] In particular embodiments, a suicide gene is utilized in conjunction with the NK cell therapy to control its use and allow for termination of the cell therapy at a desired event and/or time. The suicide gene is employed in transduced cells for the purpose of eliciting death for the transduced cells when needed. The cells of the present disclosure that have been modified to harbor one or more vectors encompassed by the disclosure that may comprise one or more suicide genes. In some embodiments, the term “suicide gene” as used herein is defined as a gene which, upon administration of a prodrug or other agent, effects transition of a gene product to a compound which kills its host cell. In other embodiments, a suicide gene encodes a gene product that is, when desired, targeted by an agent (such as an antibody) that targets the suicide gene product.

[0300] In some cases, the cell therapy may be subject to utilization of one or more suicide genes of any kind when an individual receiving the cell therapy and/or having received the cell therapy shows one or more symptoms of one or more adverse events, such as cytokine release syndrome, neurotoxicity, anaphylaxis/allergy, and/or on-target/off tumor toxicities (as examples) or is considered at risk for having the one or more symptoms, including imminently. The use of the suicide gene may be part of a planned protocol for a therapy or may be used only upon a recognized need for its use. In some cases the cell therapy is terminated by use of agent(s) that targets the suicide gene or a gene product therefrom because the therapy is no longer required.

[0301] Utilization of the suicide gene may be instigated upon onset of at least one adverse event for the individual, and that adverse event may be recognized by any means, including upon routine monitoring that may or may not be continuous from the beginning of the cell therapy. The adverse event(s) may be detected upon examination and/or testing. In cases wherein the individual has cytokine release syndrome (which may also be referred to as cytokine storm), the individual may have elevated inflammatory cytokine(s) (merely as examples: interferon-gamma, granulocyte macrophage colony-stimulating factor, IL- 10, IL-6 and TNF-alpha); fever; fatigue; hypotension; hypoxia, tachycardia; nausea; capillary leak; cardiac/renal/hepatic dysfunction; or a combination thereof, for example. In cases wherein the individual has neurotoxicity, the individual may have confusion, delirium, aplasia, and/or seizures. In some cases, the individual is tested for a marker associated with onset and/or severity of cytokine release syndrome, such as C-reactive protein, IL-6, TNF-alpha, and/or ferritin.

[0302] Examples of suicide genes include engineered nonsecretable (including membrane bound) tumor necrosis factor (TNF)-alpha mutant polypeptides (see PCT/US19/62009, which is incorporated by reference herein in its entirety), and they may be affected by delivery of an antibody that binds the TNF-alpha mutant. Examples of suicide gene/prodrug combinations that may be used are Herpes Simplex Virus-thymidine kinase (HSV-tk) and ganciclovir, acyclovir, or FIAU; oxidoreductase and cycloheximide; cytosine deaminase and 5- fluorocytosine; thymidine kinase thymidylate kinase (Tdk::Tmk) and AZT; and deoxy cytidine kinase and cytosine arabinoside. The E. coli purine nucleoside phosphorylase, a so-called suicide gene that converts the prodrug 6-methylpurine deoxyriboside to toxic purine 6- methylpurine, may be utilized. Other suicide genes include CD20, CD52, inducible caspase 9, purine nucleoside phosphorylase (PNP), Cytochrome p450 enzymes (CYP), Carboxypeptidases (CP), Carboxylesterase (CE), Nitroreductase (NTR), Guanine Ribosyltransferase (XGRTP), Glycosidase enzymes, Methionine-a,y-lyase (MET), EGFRv3, and Thymidine phosphorylase (TP), as examples.

F. Knockout or Knockdown of Endogenous Genes [0303] In certain embodiments, NK cells of the disclosure may include gene editing of the NK cells to remove 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more endogenous genes in the NK cells. In some cases the gene editing occurs in NK cells expressing one or more heterologous transgenes (e.g., CD3, TCR, etc.), whereas in other cases the gene editing occurs in NK cells that do not express a heterologous transgene but that ultimately will express one or more heterologous transgenes, in at least some cases. In particular embodiments, the NK cells that are gene edited are expanded NK cells.

[0304] In particular cases, one or more endogenous genes of the NK cells are modified, such as disrupted in expression where the expression is reduced in part or in full. In specific cases, one or more genes are knocked down or knocked out using processes of the disclosure. In specific cases, multiple genes are knocked down or knocked out in the same step as processes of the disclosure. The genes that are edited in the NK cells may be of any kind, but in specific embodiments the genes are genes whose gene products inhibit activity and/or proliferation of NK cells. In specific cases the genes that are edited in the NK cells allow the NK cells to work more effectively in a tumor microenvironment. In specific cases, the genes are one or more of NKG2A, SIGLEC-7, LAG3, TIM3, CISH, FOXO1, TGFBR2, TIGIT, CD96, ADORA2, NR3C1, PD1, PDL-1, PDL-2, CD47, SIRPA, SHIP1, ADAM17, RPS6, 4EBP1, CD25, CD40, IL21R, ICAM1, CD95, CD80, CD86, IL10R, TDAG8, CD5, CD7, SLAMF7, CD38, LAG3, TCR, beta2-microglobulin, HLA, CD73, CREB, CREM, ICER, and CD39. In specific embodiments, the TGFBR2 gene is knocked out or knocked down in the NK cells. In specific embodiments, the CISH gene is knocked out or knocked down in the NK cells. In specific embodiments, the CD38 gene is knocked out or knocked down in the NK cells.

[0305] In some embodiments, the gene editing is carried out using one or more DNA- binding nucleic acids, such as alteration via an RNA-guided endonuclease (RGEN). For example, the alteration can be carried out using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins. In general, "CRISPR system" refers collectively to transcripts and other elements involved in the expression of or directing the activity of CRISPR-associated ("Cas") genes, including sequences encoding a Cas gene, a tracr (trans-activating CRISPR) sequence (e.g., tracrRNA or an active partial tracrRNA), a tracr-mate sequence (encompassing a "direct repeat" and a tracrRNA-processed partial direct repeat in the context of an endogenous CRISPR system), a guide sequence (also referred to as a "spacer" in the context of an endogenous CRISPR system), and/or other sequences and transcripts from a CRISPR locus. Methods of utilizing a CRISPR system are well known in the art IV. Administration of Therapeutic Compositions

[0306] The CD3 -expressing NK cells and the bispecific or multi-specific antibodies are administered to an individual in need thereof, including in such a way as to be in proximity for the anti-CD3 antibody of the bispecific or multi-specific antibody to be able to bind CD3 on the CD3 -expressing NK cells. In some cases, the two components are administered separately to an individual, whereas in other cases the two components are complexed together prior to administration, such as in an ex vivo manner. In another embodiment, the NK cells express the antibodies. In some cases, the two components are not pre-complexed prior to administration, but are co-administered by any suitable route of administration, such as by co-infusion to the patient.

[0307] Embodiments of the present disclosure concern methods for the use of the compositions comprising NK cells and antibodies provided herein for treating or preventing a medical disease or disorder. The method includes administering to the subject a therapeutically effective amount of the CD3 (±TCR)-modified NK cells with the antibodies, thereby treating or preventing the disease in the subject, including reducing the risk of, reducing the severity of, and/or delaying the onset of the disease. In certain embodiments of the present disclosure, cancer or infection is treated by transfer of a composition comprising the NK cell population and corresponding antibodies. In at least some cases, because of their release of pro- inflammatory cytokines, NK cells may reverse the anti-inflammatory tumor microenvironment and increase adaptive immune responses by promoting differentiation, activation, and/or recruitment of accessory immune cell to sites of malignancy. In certain embodiments, a providing step may comprise culturing the NK cells with antibody molecules for a specific duration of time (e.g., about 5 minutes to about 24 hours or more) and storing the NK cells and the antibody molecules for a period of time (e.g., about 1, 2, 3, 4, 5 days, or greater than 5 days) prior to infusion/administration.

[0308] Cancers for which the present treatment methods are useful include any malignant cell type, such as those found in a solid tumor or a hematological tumor. Exemplary solid tumors can include, but are not limited to, a tumor of an organ selected from the group consisting of pancreas, colon, cecum, stomach, brain, head, neck, ovary, kidney, larynx, sarcoma, lung, bladder, melanoma, prostate, and breast. Exemplary hematological tumors include tumors of the bone marrow, T or B cell malignancies, leukemias, lymphomas, blastomas, myelomas, and the like. Further examples of cancers that may be treated using the methods provided herein include, but are not limited to, lung cancer (including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung), cancer of the peritoneum, gastric or stomach cancer (including gastrointestinal cancer and gastrointestinal stromal cancer), pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, various types of head and neck cancer, and melanoma.

[0309] The cancer may specifically be of the following histological type, though it is not limited to these: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; nonencapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma; ceruminous adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; paget's disease, mammary; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma w/squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; thecoma, malignant; granulosa cell tumor, malignant; androblastoma, malignant; sertoli cell carcinoma; leydig cell tumor, malignant; lipid cell tumor, malignant; paraganglioma, malignant; extra-mammary paraganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignant melanoma; amelanotic melanoma; superficial spreading melanoma; lentigo malignant melanoma; acral lentiginous melanomas; nodular melanomas; malignant melanoma in giant pigmented nevus; epithelioid cell melanoma; blue nevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma, malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mixed tumor, malignant; mullerian mixed tumor; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant; hemangiosarcoma; hemangioendothelioma, malignant; kaposi's sarcoma; hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal chondrosarcoma; giant cell tumor of bone; ewing's sarcoma; odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma, malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma; glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor; meningioma, malignant; neurofibrosarcoma; neurilemmoma, malignant; granular cell tumor, malignant; malignant lymphoma; hodgkin's disease; hodgkin's; paragranuloma; malignant lymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-hodgkin's lymphomas; B-cell lymphoma; low grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; Waldenstrom's macroglobulinemia; malignant histiocytosis; multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphoid leukemia; plasma cell leukemia; erythroleukemia; lymphosarcoma cell leukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia; myeloid sarcoma; hairy cell leukemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); acute myeloid leukemia (AML); and chronic myeloblastic leukemia.

[0310] The therapy provided herein may comprise administration of a combination of therapeutic agents, such as a first cancer therapy and a second cancer therapy. The therapies may be administered in any suitable manner known in the art. For example, the first and second cancer treatment may be administered sequentially (at different times) or concurrently (at the same time). In some embodiments, the first and second cancer treatments are administered in a separate composition. In some embodiments, the first and second cancer treatments are in the same composition. Embodiments of the disclosure relate to compositions and methods comprising therapeutic compositions. The different therapies may be administered in one composition or in more than one composition, such as 2 compositions, 3 compositions, or 4 compositions. Various combinations of the agents may be employed. Examples of therapies other than those of the present disclosure include surgery, chemotherapy, drug therapy, radiation, hormone therapy, immunotherapy (other than that of the present disclosure), or a combination thereof.

[0311] The therapeutic agents of the disclosure may be administered by the same route of administration or by different routes of administration. In some embodiments, the cancer therapy is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. In some embodiments, the antibiotic is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. The appropriate dosage may be determined based on the type of disease to be treated, severity and course of the disease, the clinical condition of the individual, the individual's clinical history and response to the treatment, and the discretion of the attending physician.

[0312] The treatments may include various “unit doses.” Unit dose is defined as containing a predetermined-quantity of the therapeutic composition. The quantity to be administered, and the particular route and formulation, is within the skill of determination of those in the clinical arts. A unit dose need not be administered as a single injection but may comprise continuous infusion over a set period of time. In some embodiments, a unit dose comprises a single administrable dose.

[0313] The quantity to be administered, both according to number of treatments and unit dose, depends on the treatment effect desired. An effective dose is understood to refer to an amount necessary to achieve a particular effect. In the practice in certain embodiments, it is contemplated that doses in the range from 10 mg/kg to 200 mg/kg can affect the protective capability of these agents. Thus, it is contemplated that doses include doses of about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, and 200, 300, 400, 500, 1000 pg/kg, mg/kg, pg/day, or mg/day or any range derivable therein. Furthermore, such doses can be administered at multiple times during a day, and/or on multiple days, weeks, or months.

[0314] In certain embodiments, the effective dose of the pharmaceutical composition is one which can provide a blood level of about 1 pM to 150 pM. In another embodiment, the effective dose provides a blood level of about 4 pM to 100 pM.; or about 1 pM to 100 pM; or about 1 pM to 50 pM; or about 1 pM to 40 pM; or about 1 pM to 30 pM; or about 1 pM to 20 pM; or about 1 pM to 10 pM; or about 10 pM to 150 pM; or about 10 pM to 100 pM; or about 10 pM to 50 pM; or about 25 pM to 150 pM; or about 25 pM to 100 pM; or about 25 pM to 50 pM; or about 50 pM to 150 pM; or about 50 pM to 100 pM (or any range derivable therein). In other embodiments, the dose can provide the following blood level of the agent that results from a therapeutic agent being administered to a subject: about, at least about, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,

29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53,

54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,

79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 pM or any range derivable therein. In certain embodiments, the therapeutic agent that is administered to a subject is metabolized in the body to a metabolized therapeutic agent, in which case the blood levels may refer to the amount of that agent. Alternatively, to the extent the therapeutic agent is not metabolized by a subject, the blood levels discussed herein may refer to the unmetabolized therapeutic agent.

[0315] Precise amounts of the therapeutic composition also depend on the judgment of the practitioner and are peculiar to each individual. Factors affecting dose include physical and clinical state of the patient, the route of administration, the intended goal of treatment (alleviation of symptoms versus cure) and the potency, stability and toxicity of the particular therapeutic substance or other therapies a subject may be undergoing.

[0316] It will be understood by those skilled in the art and made aware that dosage units of pg/kg or mg/kg of body weight can be converted and expressed in comparable concentration units of pg/ml or mM (blood levels), such as 4 pM to 100 pM. It is also understood that uptake is species and organ/tissue dependent. The applicable conversion factors and physiological assumptions to be made concerning uptake and concentration measurement are well-known and would permit those of skill in the art to convert one concentration measurement to another and make reasonable comparisons and conclusions regarding the doses, efficacies and results described herein.

V. Kits

[0317] Certain aspects of the present disclosure also concern kits comprising compositions of the invention or compositions to implement methods of the invention. In particular embodiments, the kit comprises NK cells, fresh or frozen, and that may or may not have been pre-activated or expanded. The NK cells may or may not already express one or more components of the TCR/CD3 complex. In cases wherein the NK cells do not already express one or more components of the TCR/CD3 complex, the kit may comprise reagents for corresponding transfection or transduction of the NK cells, including reagents such as vectors that express the component s), primers for amplification of the component(s), and so forth. In some cases, the NK cells may or may not also express one or more heterologous proteins as defined herein, and when they do not, the kit may comprise vectors that express the heterologous protein(s), primers for amplification of the heterologous protein(s), and so forth. [0318] Kits may comprise components which may be individually packaged or placed in a container, such as a tube, bottle, vial, syringe, or other suitable container means. Individual components (such as but not limited to NK cells, modified NK cells, means for modifying NK cells, antibodies (e.g., mono-, bi- and/or multi-specific), cytokines, etc.) may also be provided in a kit in concentrated amounts; in some embodiments, a component is provided individually in the same concentration as it would be in a solution with other components. Concentrations of components may be provided as lx, 2x, 5x, lOx, or 20x or more. In some embodiments the components are provided in separate solutions or separate containers, in some embodiments the components are provided in a single container.

VI. Examples

[0319] The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

EXAMPLE 1

PREPARATION AND EFFECTIVE USE OF CD3-EXPRESSING NK CELLS

[0320] The present example concerns cancer immunotherapeutics as a strategy to redirect the specificity of NK cells against one or more target antigens by ‘arming’ or pre-complexing them with bispecific or multi-specific antibodies, such as either prior to infusion or by coinfusing the two products separately. The NK cells are transduced with one or multiple CD3 chains, including CD3(^, CD3y, CD35 and CD3s chains and can be from any source. The cells can be expanded or non-expanded, they can be pre-activated with one or more inflammatory cytokines, such as IL 12/15/18, and/or they can be genetically modified to express one or more heterologous proteins, including, for example, engineered antigen receptors, such as chimeric antigen receptor or a TCR, and/or a cytokine gene and/or a homing/chemokine receptor.

[0321] FIG. 1 A and IB illustrate different embodiments of NK cells engineered to be utilized with bispecific or multi-specific antibodies. As shown in FIG. 1 A, in a first generation of NK cells, the cells are engineered to express CD3 that may be activated with a bispecific or multi-specific antibody, including a bispecific T cell engager (BiTE) that comprises an anti- CD3 antibody that binds heterologous CD3 expressed on the surface of the NK cells. In another embodiment, CD3 -expressing NK cells are able to be bound by a BiTE that comprises an anti- CD3 antibody, and the NK cells are also expressing one or more particular cytokines (e.g., IL- 15 and/or IL-21), resulting in increased efficacy and potency that are particularly useful for treating solid tumors. In another embodiment, the NK cells are engineered to express not only CD3 to be able to be activated by a BiTE that comprises an anti-CD3 antibody but also are utilized with a bispecific or multi-specific antibody e.g., bispecific NK cell engager, or BiKE) that comprises an antibody that binds a surface antigen naturally present on NK cells, such as CD16, CS1, CD32, CD64, CD56, NKG2D, NKG2C, DNAM, 2B4, CD2, an NCR, NKp30, NKp44, NKp46, or KIR, for example. In this manner, the NK cells respond to both NK engagers and T cell engagers. In another embodiment, the NK cells in addition to expressing CD3 to engage with T cell engagers also express an engineered antigen receptor, such as a CAR or engineered TCR.

[0322] FIG. IB illustrates different embodiments wherein the NK cells are modified to express both CD3 and a TCR. On the right, T cell TCR is illustrated having a and 0 chains with an antigen binding site wherein the TCR is complexed with CD3<^ to effect signal transduction. The T cell TCR is co-complexed with two CD3s chains, a CD36 chain, and a CD3y chain. In some embodiments, the NK cells express a TCR in which one or more of the cytoplasmic domains of any of the CD3 molecules is a heterologous intracellular domain, such as one from CD 16, NKG2D, DAP 10, DAP 12, NCR, and DNAM-1. As shown on the left of FIG. IB, the NK cells are configured to express a CD3 co-receptor component, and in one example the CD3 component is CD3s. In such a case, a standard BiTE (top left that comprises an antibody against a tumor antigen and an antibody against CD3) normally utilized with T cells that naturally express CD3 may be utilized in conjunction with the CD3 -expressing NK cells. In this particular example, the NK cells express a polypeptide that comprises the extracellular domain of CD3s (although the extracellular domains of other CD3 components may be utilized) and the extracellular domain of CD3s is linked to a transmembrane domain and/or cytoplasmic domain of another molecule, such as the transmembrane domain and/or cytoplasmic domain of CD3^, CD16, NKG2D, DAP10, DAP12, NCR, or DNAM-1, for example.

[0323] As described above, FIG. 1C. schematically depicts the generation of surface- expressible single chimeric CD3 constructs that can be used in conjunction with anti-CD3 BiTEs. As one example, the CD3 epsilon extracellular domain (ECD) is fused with CD28, CD 16, or NKG2D transmembrane (TM), and CD28, CD 16, or NKG2D intracellular domain (ICD), with or without CD3 zeta and/or DAP 10 intracellular domains. In one example, the constructs are encompassed within the Moloney murine virus-derived SFG retroviral vector backbone, which may be used with packing plasmids for viral production. In instances wherein the CD3-BiTE is used with such constructs in FIG. 1C, the antibody will bind the extracellular domain of CD3s accordingly.

[0324] Embodiments of the disclosure utilize part or all of the CD3 receptor complex. As illustrated in FIGS. 2A and 2B, the NK cells may be transfected or transduced with full length CD3zeta, CD3 gamma, CD3 delta, and CD3 epsilon. In such cases, the full length of each of CD3zeta, CD3 gamma, CD3 delta, and CD3 epsilon include the extracellular domain, the transmembrane domain, and the intracellular domain. When the different components of the receptor are expressed from the same vector, they may be configured to be produced as separate polypeptides, such as utilizing IRES or 2A elements. In any case, any expression construct may be configured to express one or more cytokines, including at least IL-15.

[0325] FIG. 4 demonstrates CD3 expression on NK cells after CMV TCR complex transduction, at day 4. The figure provides FACS plots showing CD3 expression on NK cells 4 days after CMV TCR complex transduction. Non transduced (NT) NK cells (CD56+ CD3-) serve as a negative control and T cells (CD3+ CD56-) serve as a positive control.

[0326] FIG. 5 demonstrates TCR expression on NK cells after CMV TCR complex transduction of NK, day 4. In particular, provided are FACS plots showing TCRa/b expression on NK cells 4 days after CMV TCR complex transduction. Non transduced (NT) NK cells (CD56+ CD3-TCRa/b-) serve as a negative control and T cells (CD3+TCRa/b+ CD56-) serve as a positive control.

[0327] FIG. 6 shows TCR/CD3 expression on NK after CMV TCR complex transduction, day 6. Specifically, FACS plots show dual CD3 and TCRa/b expression on NK cells 6 days after CMV TCR complex transduction. Non transduced (NT) NK cells (CD56+ CD3-TCRa/b- ) serve as a negative control and T cells (CD3+TCRa/b+ CD56-) serve as a positive control.

[0328] In FIG. 7, shown are the binding of CD3-CD19 BiTE on NK cells through the CD3/TCR at different concentrations. Specifically, the various cells (non-transduced (NT) NK cells, T cells, or the three different NK-TCR cells) were incubated with blinatumomab a CD3- CD19 bispecific engager (BiTe) for one hour at 37°C using two different concentrations (0.5 pg/pl or 4 pg/pl). Then, a biotin-labeled CD19 antigen (CD19 CAR Detection Reagent from Miltenyi Biotech™) was added for 20 min followed by an anti-biotin antibody for 15 min at room temperature. This strategy was used to detect any BiTe engaged with a CD3+ cell. The Histograms in FIG. 7 show the level of CD19 binding to CD3-CD19 bispecific engager (BiTe) that correlates with CD3 expression on NK-TCR and T cells.

[0329] FIG. 8 shows NK-TCR cytokine production after stimulation with a plate-bound CD3 antibody. In particular, CD3-OKT3 clone 20 pg/ml was incubated overnight in flat bottom 96-well plates at 4°C to form a plate-bound antigen. The next day, T cells or NK cells (NT or TCR-transduced) were added to the wells for 4 hrs and with Brefeldin A (that prevents the cytokine from being released, trapping it in the cytoplasm such that it can be detected by intracellular cytokine staining). They were then harvested for surface and intracellular staining to assess cytokine production and degranulation (TNFa and CD107a). FACS plots in FIG. 8 show TNFa and CD 107a double-positive populations in NK cells transduced with TCR. Nontransduced (NT) NK cells (CD56+ CD3-) serve as a negative control and T cells (CD3+ CD56- ) serve as a positive control.

[0330] FIG. 9 demonstrates phosphorylation of CD3^ in NK TCR/CD3 cells after crosslinking CD3. The various cells tested included non-transduced (NT) NK cells; nontransduced (NT) T cells, or three different CD3-TCR transduced NK cells (where CD1, CD2, or CD3 represent different donors). Each of the NK cell groups were transduced with CD3ZFLGDEFL15 (see FIGS. 2A and 2B). The NK cells were incubated with CD3 OKT3 clone (Miltenyi Biotech™, 130-093-387) at 20 pg/ml concentration for 20 min on ice. Cells were then cross-linked with Fab2 IgGl antibody for various time points and stained to check for CD3z phosphorylation. This analysis of CD3^ is useful because, as an internalization signal from the surface, it would only be able to be crosslinked with a CD3 monoclonal antibody if the NK cells expressed it. NK cells that are not transduced with CD3 will not show any phosphorylation or activation after the stimulation. [0331] NK cells transduced with CD3-TCR also show basal level of tonic signaling, which increases upon stimulation with CD3 OKT3 and is similar to T cells, while non-transduced NK cells did not show any CD3^ phosphorylation neither at basal nor upon CD3 OKT3 stimulation. [0332] FIG. 10 shows that pre-culturing CD3-CD19 BiTEs with TCR/CD3-expressing NK cells increased its killing activity against Raji cells. NK cells were either transduced with CD3- TCR#1 (CD3ZFLGDEFL15 (see FIGS. 2A and 2B)) or CD3-TCR#2 (Z2, also called CD3ZGDEFL8SP21CD8, that includes full length CD3^, full length CD3y, full length CD36, and full length CD3 s linked to membrane bound IL21 (with CD8 transmembrane domain for the membrane bound IL21). NK cells transduced with the CD3/TCR constructs or nontransduced NK cells were loaded with Blinatumumab and incubated for 1 hour and washed with PBS. They were then co-cultured with CD 19+ B cell lymphoma cells at different Effector celkTarget cell ratios (FIG. 10A is a 1 : 1 ratio, and FIG. 10B is a 1 :5 ratio) for various time points. As utilized herein, Effector cells are the CD-3-TCR NK Cells, and Target cells are the Raji cells. Blinatumumab-loaded CD3-TCR transduced NK cells showed enhanced anti-tumor activity compared to Blinatumumab-loaded non-transduced NK cells or CD3/TCR transduced NK cells, but not loaded with Blinatumomab at both E:T ratios.

EXAMPLE 2 NY-ESO TCRS IN NK CELLS

[0333] The present examples concern generation and use of NY-ESO TCRs in NK cells. In FIG. 11, there is one example for production of the cells. The schematic overview shows one case wherein the NK cells are first transduced with the uTNK 15 construct that incorporates signaling domains from the CD3 complex, NK costimulatory molecules and IL- 15, followed by a second transduction step that introduces the TCR molecule, thus generating NK cells that co-express CD3 and NK signaling molecules, IL-15, and a TCR complex. In one embodiment, NK cells were derived from cord blood and were expanded with irradiated (100 Gy) universal antigen presenting cells (uAPC) feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL-2 (200 U/ml) in complete media. To generate a universal T cell-like NK cell (uTNK15 cells) that can secrete IL-15, NK cells were purified and transduced with a retroviral construct containing a CD3 complex with NK co-stimulatory molecules and an IL-15 gene 4 days after isolation. Forty-eight hours after the initial transduction, NK cells expressing uTNK15 were then transduced with a TCR targeting an antigen of choice.

[0334] Expression of NY-ESO TCR on NK cells transduced with uTNK15 is shown in FIG. 12. NK cells were derived from cord blood and were expanded with irradiated (100 Gy) universal antigen presenting cells (uAPC) feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL-2 (200 U/ml) in complete media. To generate a universal T cell-like NK cell that can secrete IL-15, NK cells were purified and transduced with a retroviral construct containing a CD3 complex with NK co-stimulatory molecules and an IL-15 gene 4 days after isolation. Forty-eight hours after the initial transduction, uTNK15 cells were then transduced with a TCR complex targeting an antigen of choice. Forty-eight hours later, flow cytometry was used to assess the expression of CD3 and NY-ESO TCR on the various uTNKl 5 constructs. Non transduced (NT) NK cells served as negative control. CD3 and NY-ESO TCR were highly expressed on all uTNKl 5 cells compared to NT NK cells. The number of tumor specific TCR molecules expressed on TCR engineered NK cells using the various TCR constructs are provided in FIG. 13, and NT NK cells were used as negative control.

[0335] FIG. 14 demonstrates NY-ESO TCR expression on non-transduced and transduced T cells. T cells were isolated from cord blood (the same donor as NK cells to serve as a paired positive control) and were activated with CD3/CD28 microbeads at a concentration of 25 pl/ 1 million for 48 hours in RPMI complete media. T cells were then transduced with a retroviral construct containing NY-ESO TCR. Forty-eight hours after transduction, flow cytometry revealed that NY-ESO TCR was highly expressed on transduced T cells compared to nontransduced T cells.

[0336] NK cells transduced with NY-ESO TCR kill NY-ESO peptide-pulsed target cells in a dose-dependent manner (FIG. 15). Chromium ⁵¹CR killing assay was performed 7 days following TCR transduction to determine the killing capacity of TCR-engineered NK and T cells against LCL cells loaded with different concentrations of NY-ESO peptide for 2 hours. NY-ESO TCR transduced uTNKl 5 cells show enhanced killing of peptide-pulsed LCL cells compared to non-transduced NK cells. NY-ESO TCR transduced T cells show enhanced killing of peptide-pulsed LCL cells compared to non-transduced T cells.

[0337] FIG. 16 shows that NY-ESO is expressed endogenously on myeloma, sarcoma, and melanoma cell lines. Flow cytometry was used to determine the expression of NY-ESO on U266 (myeloma), Saos-2 (Sarcoma), and A375 (melanoma) cell lines. U266, Saos-2, and A375 cell lines showed higher levels of NY-ESO expression compared to the Raji cell line which served as negative control.

[0338] NY-ESO TCR-transduced T cells kill NY-ESO expressing tumor targets at higher E:T ratios (FIG. 17). Chromium ⁵¹CR killing assay was performed 7 days following TCR transduction to determine the killing capacity of NY-ESO TCR-engineered T cells against NY- ESO expressing myeloma, osteosarcoma and melanoma cell lines. NY-ESO TCR transduced T cells show enhanced killing of NY-ESO positive cell lines compared to non-transduced T cells.

[0339] FIG. 18 demonstrates that NY-ESO TCR transduced NK cells kill NY-ESO expressing tumor targets even at low E:T ratios. Chromium ⁵¹CR killing assay was performed 7 days following TCR transduction to determine the killing capacity of NY-ESO TCR- engineered NK cells against NY-ESO-expressing myeloma, osteosarcoma and melanoma cell lines. NY-ESO TCR-transducedNK cells show enhanced killing of NY-ESO positive cell lines compared to non-transduced NK cells even at very low effector: target ratios.

[0340] FIG. 19 shows that NY-ESO transduced NK cells have a similar phenotype to NT NK cells. CytoF imaging revealed that non-transduced NK cells and NY-ESO TCR transduced uTNK15 cells share a similar phenotype. FIG. 19A shows a u-map plot with similar clusters, and FIG. 19B shows a heat map with similar expression of various markers on NT and NY- ESO TCR transduced uTNK15 cells.

[0341] FIG. 20 provides a table representing the percentage of CD3+ and CD3+TCR+ NK cells in each uTNK15 product. Flow cytometry was used to assess the composition of single positive CD3 NK cells (CD3+) and double positive CD3/TCR NK cells (CD3+TCR+). Non transduced NK cells are comprised of less than 1% CD3+ and CD3+TCR+ NK cells, while the TCR transduced uTNK15 cell products are comprised of over 60% CD3+ and over 25% CD3+TCR+ NK cells.

[0342] FIG. 21A provides FACS plots that show successful CD3 expression on NK cells 4 days after transduction with TCR constant alpha-beta (TCRCab; TCR6 construct). Non transduced (NT) NK cells (CD56+ CD3-) serve as negative control. In FIG. 21B, NT NK and uTNK15 NK cells were incubated with Blinatumumab, a CD3-CD19 bispecific engager (BiTe), for one hour at 37°C using 10 pg/pl. Then, a biotin-labeled CD19 antigen (CD19 CAR Detection Reagent from Miltenyi Biotech™) was added for 20 min, followed by an anti-biotin antibody for 15 min at room temperature. This strategy was used to detect any BiTe engaged with a CD3+ cell. The histograms in this figure are showing the level of CD19 binding to CD3- CD19 bispecific engager (BiTe) that correlates with CD3 expression on uTINK15 NK cells. In FIG. 21C, CD3/TCR transduced or non-transduced NK cells were loaded with Blinatumumab and incubated for 1 hour and washed with PBS. They were then co-cultured with LCL cells at different E:T ratios (A.1:1,B.1:5) for various time points. Blinatumumab-loaded CD3-TCR transduced NK cells showed enhanced anti-tumor activity compared to Blinatumumab-loaded non-transduced NK cells or CD3/TCR transduced NK cells but not loaded with Blinatumumab at both E:T ratios. EXAMPLE 3

NY-ESO TCRS IN CD3 EXPRESSING NK CELLS IN VIVO

[0343] As shown in FIGS. 22A-22C, NK cells comprising constructs described herein were tested in-vivo and found to robustly inhibit tumor growth. Shown in FIG. 22A is a schematic outlining the experimental procedure performed. In brief, NSG mice were irradiated with 300 cGy on day -1, then on day 0 individual mice received tail vein injections of 0.5 x 10⁶ U266- B1 cells (a myeloma cell line that expresses both HLA-A2 and NY-ESO antigens) that were transduced with FireFlyluciferase (FFluc), on day 3 mice were infused with 5 x 10⁶ effector cells (NY-ESO TCR NK cells with WT, #A, or #B UT-NK15-NY ESO TCR constructs respectively; WT refers to wild type CD3 molecules with IL- 15; #A refers to CD3-CD28 with IL-15 (e.g., UT-NK15-28); and #B refers to CD3 -DAP 10 with IL- 15 (e.g., UT-NK15-DAP10); or NY-ESO TCR T cells), animals were then monitored over time and sacrificed as appropriate (N = 5 mice per group). FIG. 22B displays the results of the monitoring of the experiment described in FIG. 22A as a function of bioluminescent imaging over time (displayed are representative images from day 1, day 7, day 14, and day 21 respectively). FIG. 22C is a graphical quantification of the bioluminescence average radiance displayed in FIG. 22B, the Y axis denotes average radiance in p/s/cm²/sr, while the X axis denotes time.

[0344] As shown in FIGS. 23A-B the in vitro activity of effector cells (e.g., NK cells or T cells) comprising NY-ESO targeted TCRs and UT-NK15 constructs was tested. FIG 22A are images of spheroids formed by osteosarcoma tumor cell line Saos-2 that were used to test the activity of NY-ESO1 -specific TCR expressing NK and T cells cytotoxicity. Saos-2 cells were stably transduced to express GFP; 10,000 of these cells were seeded per well in a 96 well plate overnight and 40,000 of NK or T cells were then added. Images of the coculture were scanned over time and analyzed by the IncuCyte® cell analysis system. Shown in FIG. 22B is a graph displaying the percentage of cytotoxicity (Y axis) for effector cells captured from representative images after 3 days of co-culture. NK cells were co-transduced with NY-ESO- TCR, and the UT-NK15 signaling complex co-expressing different co-stimulatory molecules fused to the CD3(^ signaling chain (e.g., UTNK-15-28, orUTNK-15-DAP10). T cells were only transduced with NY-ESO TCR. Abbreviations in the graph are as follows: 28 = CD3(^ fused to a CD28 co-stimulatory domain; 10 = CD3(^ fused to a Dap 10 co-stimulatory domain; 8 = CD8 alpha/beta co-receptor as part of the NY ESO TCR construct; wo IL- 15 = the construct only contains CD3 zeta, epsilon, gamma and delta without co-stimulation or IL-15. The best in vitro cytotoxicity was observed with TCR NK cells expressing UTNK15 with CD28, or DAP10 costimulatory domains fused to CD3(^ (e.g., UTNK-15-28, or UTNK-15-DAP10; SEQ ID NO: 121 and SEQ ID NO: 119 respectively) when compared to NK cells transduced with CD3 complex only or the UT-NK15 without a co-stimulatory domain. The addition of the CD8 alpha/beta coreceptor to the TCR did not significantly improve on the cytotoxicity of NK or T cells.

[0345] As shown in FIGS. 24A-D the in vivo activity of effector cells (e.g., NK cells or T cells) comprising NY-ESO targeted TCRs and UT-NK15 constructs was tested. FIG. 24A depicts a plan for an in vivo study to test the activity of different NY ESO TCR transduced NK and T cells. The plan was performed, wherein ten week old NSG mice were irradiated (300 cGy) and the next day they were injected with 500,000 U266 cells (HLA-A2 positive, NY- ESO-expressing myeloma cell line) via the tail vein. Three days later, the mice received 5 million TCR transduced T or TCR-transduced NK cells. Mice were then monitored for tumor control by BLI imaging. Shown in FIG. 24B are said BLI imaging results of the test outlined and performed according to FIG. 24 A. Mice were injected with U266 tumor cells only, or also with T cells transduced with NY-ESO-specific TCR, or also with NK cells co-transduced with NY-ESO TCR and UT-NK15 with CD3^ fused to CD28 (labelled as NY-ESO NK UT-NK15 CD28 or NY-ESO TCR UTNK-15 CD28 NK cells). Shown in FIG. 24C are quantifications of region of interest average radiance intensity for the animals tested according to FIG. 24A and imaged in FIG. 24B. Shown in FIG. 24D is a graph depicting the cohort survival curves for the aforementioned animals. The results showed that NY ESO TCR T and NY-ESO TCR UTNK- 15-CD28 NK cells mediated strong antitumor activity in vivo.

[0346] As shown in FIG. 25 the in vivo activity of effector cells (e.g., NK cells) comprising NY ESO TCR and CD3 complex with or without IL- 15 was tested. NSG mice were irradiated (300 cGy) and the next day were injected with 500,000 U266 cells (HLA-A2 positive, NY- ESO-expressing myeloma cell line) via the tail vein. Three days later, mice received 5 million TCR transduced T or NK cells. Mice were monitored for tumor control by BLI imaging. NK cells were transduced with NY-ESO-specific TCR, and co-transduced with CD3 complex without IL-15 or with UT-NK15 expressing CD3(^ fused to CD28 (UT-NK15-28) or CD3(^ fused to DAP10 (UT-NK15-DAP10) co-stimulatory molecules, with or without expression of CD8 alpha/beta co-receptors. The results showed that absence of IL- 15 resulted in a reduced anti-tumor activity in vivo.

[0347] Together these results showed that effector cells (e.g., NK cells) comprising constructs described herein (e.g., NY-ESO TCR constructs and/or CD3 constructs such as UT- NK15 or modified versions thereof, e.g., UT-NK-15-28 or UT-NK15-DAP10) were sufficient to robustly inhibit tumor growth in vivo. EXAMPLE 4

PRAME TCRS IN CD3 EXPRESSING NK CELLS IN VITRO

[0348] As shown in FIGS. 26A-C, NK cells comprising constructs targeting Preferentially Expressed Antigen In Melanoma (PRAME) antigen described herein were tested in-vitro and found to robustly inhibit tumor cell growth. FIG. 26 A shows the expression of both UT-NK15 (x-axis, CD3) and PRAME-specific TCRs (y-axis, TCR) in NK cells (TCR clones 46, 54, or DSK3 respectively), or the expression of PRAME-specific TCRs in T cells transduced with the same (TCR clones 46 or 54). PRAME-specific TCR expression on NK cells was confirmed using antibodies against the TCR and against CD3. Expression of PRAME-specific TCR in T cells was confirmed by tetramer staining using the 46/54 peptide/MHC-specific tetramer. FIG. 26B shows the in vitro cytotoxicity of NK cells expressing a PRAME-specific TCR against the U266 myeloma cell line. IncuCyte® live cell imaging was used to measure the cytotoxicity of T cells transduced with PRAME-specific TCR and NK cells transduced with UT-NK15 and PRAME-specific TCR against U266 myeloma cells. GFP-expressing U266 cells were cocultured with PRAME-specific TCR expressing T cell or NK cells at 1 : 1 effector : target ratio (50,000 effector and 50,000 target cells were seeded in each well of a 96 well plate). A reduction in GFP expression indicated cell death. After 26 hours, a second round of 50,000 tumor cells was added (noted as “rechallenging”) to each well for the tumor rechallenge assay. NK cells expressing UT-NK15 and PRAME-specific TCR clone 46 or PRAME-specific TCR clone 54 exerted the best anti-tumor activity upon rechallenge with U266 cells and displayed superior cytotoxicity when compared to control T cells transduced with PRAME-specific TCR clones 46 or 54 respectively. FIG. 26C shows the in vitro cytotoxicity of NK cells expressing a PRAME-specific TCR against the UA375 melanoma cell line. IncuCyte® live cell imaging was used to measure the cytotoxicity of T cells transduced with PRAME-specific TCR and NK cells transduced with UT-NK15 and PRAME-specific TCR against UA375 melanoma cells. GFP-expressing UA375 cells were co-cultured with PRAME-expressing T cell or NK cells at 1 : 1 effector : target ratio (50,000 effector and 50,000 target cells were seeded in each well of a 96 well plate). A reduction in GFP expression indicated cell death. After 26 hours, a second round of 50,000 tumor cells was added to each well for the tumor rechallenge assay. Open symbols represent T cells, while closed symbols represent NK cells. NT = non-transduced. NK cells expressing UT-NK15 and PRAME-specific TCR clone 46 (TCR-46), PRAME-specific TCR clone 54 (TCR-54), or PRAME-specific TCR clone DSK3 (DSK) exerted strong antitumor activity upon rechallenge with UA375 cells, and displayed superior cytotoxicity when compared to control T cells transduced with PRAME-specific TCR clones 46, 54, or DSK3 respectively.

[0349] Together these results show that effector cells (e.g., NK cells) comprising constructs described herein (e.g., PRAME-specific TCR constructs) were sufficient to robustly inhibit tumor growth in vivo. Furthermore, NK cells comprising CD3 constructs described herein coupled with PRAME-specific TCR constructs displayed increased cytotoxicity when compared to T cell control cells comprising the same TCR constructs, particularly in cases of continuous and/or rechallenge by tumor cells.

EXAMPLE 5

TCRS IN CD3 EXPRESSING NK CELLS IN VIVO

[0350] NK cells comprising constructs described herein are tested in-vivo and robustly inhibit tumor growth. Experiments are performed according to schematics and experimental procedures described herein. In brief, NSG mice are irradiated (e.g., with about 300 cGy) on day -1, then on day 0 individual mice receive tail vein injections of cancer cells (e.g., 0.5 x 10⁶ cells e.g., cells expressing (naturally and/or transduced with) an antigen described herein) that are transduced with an appropriate marker (e.g., FireFlyluciferase (FFluc)), on day 3 mice are infused with effector cells transduced with a transgenic TCR (e.g., TCR constructs comprising gamma/delta TCR chains, alpha/beta invariant TCR chains, and/or alpha/beta TCR chains, e.g., targeting antigens described herein, e.g., NY-ESO, Tyrosinase, MAGEA3, MAGEA4, HPV E7, WT1, PRAME, gplOO, MART-1, CD19, CD20, glycolipids, etc.) and with or without other constructs described herein (e.g., with about 5 x 10⁶ TCR and/or iTCR NK cells with a UT- NK15 construct with or without IL 15, with or without CD3 fusion to a costimulatory molecule, and/or with or without additional control constructs). Animals are then monitored over time and sacrificed as appropriate. Results of the monitoring of the experiment described above are recorded, e.g., as a function of bioluminescent imaging over time (e.g., on day 1, day 7, day 14, day 21, etc).

[0351] The in vitro activity of effector cells (e.g., NK cells or T cells) comprising TCR(s) (e.g., TCR constructs comprising gamma/delta TCR chains, alpha/beta invariant TCR chains, and/or alpha/beta TCR chains, e.g., targeting antigens described herein, e.g., NY-ESO, Tyrosinase, MAGEA3, MAGEA4, HPV E7, WT1, PRAME, gplOO, MART-1, CD 19, CD20, glycolipids, etc.) and UT-NK15 constructs are tested. Spheroids formed by an appropriate tumor cell line(s) comprising an antigen of interest (e.g., 0.5 x 10⁶ cells e.g., cells expressing (naturally and/or transduced with) an antigen described herein) are used to test the activity of specific TCR and/or iTCR expressing NK and/or T cells cytotoxicity. Cancer cells are stably transduced to express an appropriate marker (e.g., GFP, FFluc, etc.); a number of these cells (e.g., about 10,000) are seeded per well in a 96 well plate overnight and a number of effector cells (e.g., about 40,000) are then added. Images of the coculture are scanned over time and analyzed by an appropriate system (e.g., an IncuCyte® cell analysis system). The percentage of cytotoxicity for effector cells are captured from representative images after a number of days (e.g., 1 day, 3 days, 7 days, etc.) of co-culture. NK cells are co-transduced with antigen targeting TCRs and/or iTCRs, and UT-NK15 signaling complex co-expressing different costimulatory molecules fused to the CD3(^ signaling chain (e.g., UTNK-15-28, or UTNK-15- DAP10). Appropriate control cells are transduced with appropriate constructs described herein. Superior in vitro cytotoxicity is observed with TCR NK and/or iTCR NK cells expression UTNK15 with CD28, or DAP10 costimulatory domains fused to CD3(^ (e.g., UTNK-15-28, or UTNK-15-DAP10; e.g., SEQ ID NO: 121 and SEQ ID NO: 119 respectively) when compared to NK cells transduced with CD3 complex only or UT-NK15 without a co-stimulatory domain. [0352] The in vivo activity of effector cells (e.g., NK cells or T cells) comprising antigen specific TCRs (e.g., TCR constructs comprising gamma/delta TCR chains, alpha/beta invariant TCR chains, and/or alpha/beta TCR chains, e.g., targeting antigens described herein, e.g., NY- ESO, Tyrosinase, MAGEA3, MAGEA4, HPV E7, WT1, PRAME, gplOO, MART-1, CD 19, CD20, glycolipids, etc.) and UT-NK15 constructs are tested. Assays for in vivo analysis of effector cells (e.g., NK cells or T cells) comprising engineered constructs are performed similar to experimental plans described in FIG. 24. In brief, appropriately aged NSG mice (e.g., ten week old NSG mice) are irradiated (e.g., with about 300 cGy) and the next day they are injected with tumor cells comprising the target antigen of interest (e.g., about 500,000 cells; e.g., naturally expressing and/or transduced with an antigen described herein) via the tail vein. Three days later, the mice receive an effector cell bolus (e.g., about 5 million TCR and/or iTCR transduced T and/or TCR and/or iTCR transduced NK cells). Mice are then monitored for tumor control (e.g., by BLI imaging). Average radiance for regions of interest are measured and quantified, animals comprising test constructs comprising TCRs and/or iTCRs targeting an antigen of interest and UT-NK15 constructs with or without CD3 fusions and/or IL-15 expression display improved survival relative to control animals and/or a reduction in average radiance. The results show that TCR UTNK-15 NK cells mediate strong antitumor activity in vivo.

[0353] The in vivo activity of effector cells (e.g., NK cells) comprising TCR (e.g., TCR constructs comprising gamma/delta TCR chains, alpha/beta invariant TCR chains, and/or alpha/beta TCR chains, e.g., targeting antigens described herein, e.g., NY-ESO, Tyrosinase, MAGEA3, MAGEA4, HPV E7, WT1, PRAME, gplOO, MART-1, CD 19, CD20, glycolipids, etc.) and CD3 complex with or without IL-15 are tested. NSG mice are irradiated (e.g., with about 300 cGy) and the next day are injected with tumor cells expressing an antigen of (e.g., about 500,000 cells; e.g., naturally expressing and/or transduced with an antigen described herein) via the tail vein. Three days later, mice receive an effector cell bolus (e.g., about 5 million TCR and/or iTCR transduced T cells, and/or TCR and/or iTCR transduced NK cells). Mice are monitored for tumor control (e.g., by BLI imaging). NK cells are transduced with antigen-specific TCRs and/or glycolipid-specific iTCRs, and co-transduced with CD3 complex without IL-15 or with UT-NK15 expressing CD3(^ fused to CD28 (UT-NK15-28) or CD3(^ fused to DAP10 (UT-NK15-DAP10) co-stimulatory molecules, with or without expression of CD8 alpha/beta co-receptors. The results show that absence of IL-15 results in a reduced antitumor activity in vivo.

[0354] Together these results show that effector cells (e.g., NK cells) comprising constructs described herein (e.g., TCR and/or iTCR constructs, and/or CD3 constructs such as UT-NK15 or modified versions thereof, e.g., UT-NK-15-28 or UT-NK15-DAP10) are sufficient to robustly inhibit tumor growth in vivo.

EXAMPLE 6

INVARIANT TCRS IN CD3 EXPRESSING NK CELLS IN VITRO

[0355] As shown in FIGS. 27A-27C, invariant Natural Killer T-cell Receptors (iTCRs) were transduced into cord blood derived NK cells, and the iTCR expression profile was determined. NK cells were derived from cord blood (three donors, CD4, CD8, and DN respectively), and were expanded with irradiated (100 Gy) universal antigen presenting cells (uAPC) feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL-2 (200 U/ml) in complete media. To create a universal T cell-like NK cell that could secrete IL- 15 and signal via CD3 upon CD3 antibody engagement, NK cells were purified and co-transduced with a retroviral construct containing a CD3 complex with NK co-stimulatory molecules and an IL- 15 gene, and with an invariant Natural Killer T Cell receptor (iTCR) (alpha and beta chains) on day 5. Following transduction, cells underwent another round of expansion with uAPCs. As shown in FIG. 27A, exemplary flow cytometry plots showed expression of three different iTCR pairs in transduced NK cell populations, with each population expressing iTCRa (SEQ ID NO: 295), and either iTCRp chain 1 (SEQ ID NO: 297), iTCRp chain 2 (SEQ ID NO: 299), or iTCRP chain 3 (SEQ ID NO: 301), and CD3. Confirmation of iTCR expression was determined using antibodies specific for iTCR target Va24 and Vpi 1 regions (y and x axis respectively). To prepare cells for analysis, NK cells were isolated from cord blood and pre-treated with IL15, IL18, and IL12 cytokine cocktail on day -1. The next day (day 0) irradiated uAPC feeder cells were added to stimulate and expand the NK cells. Transduction was done on day 5 with both uTNK15 and the exampled iNKT iTCR retroviral compositions. iTCR expression on NK cells was confirmed with antibodies specific for iTCR that targets Va24 and Vpi l regions. As shown in FIG. 27B, exemplary flow cytometry plot results showed binding of Blinatumomab (y axis) to three different CD3/iTCR complex (comprising iTCRl, iTCR2, and iTCR3, respectively) expressing NK cells derived from three donors (cell donor 4 (CD4), cell donor 8 (CD8), and cell donor N (CDN), respectively). Blinatumomab was pre-loaded on the 3 different iTCR-expressing NK cells by incubation of NK cells with Blinatumomab (4 pg/pl) for 1 hour at 37 °C. The cells were then washed once. The binding of Blinatumomab on CD3 was confirmed by flow cytometry using the CAR19 detection kit from Miltenyi Biotech™, which uses the CD 19 protein to detect Blinatumomab (y axis) binding on NK cell surface. As shown in FIG. 27C IncuCyte® live cell imaging was used to measure the cytotoxicity of NK cells pre- loaded with Blinatumomab (as detailed above), co-transduced with iTCR constructs and UT- NK15, against GFP-expressing Raji cells. GFP-expressing Raji cells were co-cultured with CD3/iTCR-expressing NK cells at 3:1 effector to target ratios (150,000 effector and 50,000 target cells were seeded in each well of a 96 well plate). T cells and iNKT cells were used as positive and negative controls, respectively. A reduction in GFP expression indicated cell death. Three NK donor lines were utilized, CB152, CB153, and CB154, respectively, transduced with iTCRs (iTCRP chain 1, 2, or 3 (SEQ ID NOs: 297, 299, or 301, respectively) and iTCRa (SEQ ID NO: 295)) or left non-transduced (NT) (as negative control).

[0356] The results depicted in FIG. 27 show that iTCRs can be expressed in cord blood derived NK cells, and that these iTCRs can be utilized to stabilize the expression of transgenic CD3. Furthermore, that the transgenic CD3 can be bound by CD3 targeting antibodies, such as Blinatumomab, to create antibody loaded CD3/iTCR-expressing NK cells. Loading of CD3/iTCR-expressing NK cells with an antibody (e.g., Blinatumomab) rendered the cells highly cytotoxic against tumor cells (e.g., Raji cells), with residual tumor cell numbers in the trials being lower for CD3/iTCR expressing NK cells when compared to control cells such as T cells, iNKT cells, and/or non-transduced NK cells.

[0357] As shown in FIGs. 28A-28C, new iTCRP sequences were obtained from cord blood derived iNKT cells, these iTCRP chains (in addition to iTCRa) were successfully transgenically expressed in cord blood derived NK cells, and facilitated improved cytotoxicity of the NK cells against tumor cells. As depicted in FIG. 28A, iNKT cells from five cord blood donors were identified and shown to express both iTCRp and iTCRa. The iNKT cells were isolated from 5 cord blood donors using the iNKT isolation kit from Miltenyi Biotech™. Isolated cells were stimulated and expanded with irradiated cord blood PBMC (40 Gry) in the presence of 100 ng/ml of alpha-Galactosylceramide and 200 U/ml of IL-2. On day 7 after iNKT expansion, the purity of iNKT cultures was confirmed by iNKT specific antibodies against Va24 and Vpi l. Total mRNA was extracted from iNKT cells, and cDNAs of the VP-DJ regions were cloned and sequenced (100 individual clones). Shown in FIG. 28B are VP-DJ nucleotide sequences of cloned iTCRp chains. As shown in FIG. 28C transgenic expression of Va24 and CD3 in transgenic CD3/iTCR expressing NK cell populations from three cord blood donors was confirmed. The NK cells were co-transduced with iTCRs (eight randomly selected iTCRp clones with coding sequences represented by SEQ ID NOs: 300, 302, 304, 306, 308, 310, 312, or 314, representing clones 3, 18, 24, 51, 56, 76, 93, and 96 respectively, and iTCRa coding sequence represented by SEQ ID NO: 292) and UT-NK15 on day 5. iTCR expression was confirmed by flow cytometry using iTCR specific antibodies against Va24 and antibodies against CD3. The cytotoxicity of these transduced cells was then tested. As shown in FIG. 28D and 28E, IncuCyte® live cell imaging was utilized to measure of cytotoxicity of NK cells cotransduced with iTCRs (as described in FIG. 28B) and UT-NK15, and pre-loaded with Blinatumomab (1 hour at 37 °C), against GFP-expressing Raji cells at 3 : 1 target to effector cell ratios (150,000 effector and 50,000 target cells were seeded in each well of a 96 well plate). T cells were used as positive control and non-transduced (NT) cells were utilized as negative controls. In general, the cytotoxicity of T cells expressing transgenic TCRs was found to be the same as cord blood derived in iNKT cells expressing transgenic TCRs. A reduction in GFP expression indicated tumor cell death. iTCR/CD3 expressing NK cells displayed significantly greater tumor cell cytotoxicity, as evidenced by lower relative residual tumor cell populations. [0358] The results depicted in FIG. 28 show that newly isolated iTCRs can be expressed in cord blood derived NK cells, and that these iTCRs can be utilized to stabilize the expression of transgenic CD3. Furthermore, that the transgenic CD3 can be bound by CD3 targeting antibodies, such as Blinatumomab, to create antibody loaded CD3/iTCR-expressing NK cells. Loading of CD3/iTCR-expressing NK cells with an antibody (e.g., Blinatumomab) rendered the majority of CD3/iTCR-expressing NK cells highly cytotoxic against tumor cells (e.g., Raji cells), with residual tumor cell numbers in the trials being lower for CD3/iTCR expressing NK cells when compared to control cells such as T cells and/or non-transduced NK cells.

* * * [0359] All of the methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Claims

WHAT IS CLAIMED IS:

1. An engineered NK cell modified to express, a) part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3 b) part or all of a single chain or any combination of invariant NK T cell receptor (iTCR) alpha (iTCRa) chain or beta (iTCR0) chain; and c) a cytokine selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, IL-7, GMCSF, and a combination thereof.

2. The engineered NK cell of claim 1, wherein the NK cells are modified to express part or all of CD36, two of CD3s, CD3y, and/or CD3^.

3. The engineered NK cell of claim 1, wherein any one or more of CD36, CD3s, CD3y, and/or CD3<^ are heterologously linked to one or more intracellular signaling domains.

4. The engineered NK cell of claim 3, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and a combination thereof.

5. The engineered NK cell of claim 3, wherein the intracellular signaling domain comprises a DAP 10 intracellular signaling domain.

6. The engineered NK cell of claim 5, wherein the intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 115.

7. The engineered NK cell of claim 3, wherein the intracellular signaling domain comprises a CD28 intracellular signaling domain.

8. The engineered NK cell of claim 7, wherein the intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 116.

9. The engineered NK cell of claim 3, wherein the intracellular signaling domain comprises a DAP 10 and CD28 intracellular signaling domain.

10. The engineered NK cell of claim 9, wherein the intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 117.

11. The engineered NK cell of claim 1, wherein the iTCRa chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 293, and the iTCR0 chain comprises an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315.

12. The engineered NK cell of claim 1, wherein the iTCRa chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 293, and the iTCR0 chain comprises an amino acid sequence at least 90% identical to SEQ ID NOs: 301, 307, 311, or 313.

13. The engineered NK cell of claim 1, wherein the cell is modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT-NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122).

14. The engineered NK cell of claim 1, wherein the NK cells are derived from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or a combination thereof.

15. The engineered NK cell of claim 1, wherein the NK cells are primary NK cells, and are not derived from stem cells and/or induced pluripotent stem cells (iPSCs).

16. The engineered NK cell of claim 1, wherein the NK cells are complexed to one or more antibodies.

17. The engineered NK cell of claim 16, wherein the one or more antibodies are one or more bispecific or multi-specific antibodies, wherein at least one of the bispecific or multi-specific antibodies comprises an anti-CD3 antibody linked.

18. The engineered NK cell of claim 16, wherein the antibody is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B.

19. The engineered NK cell of claim 16, wherein the antibody is Blinatumomab.

20. The engineered NK cell of claim 16, wherein the NK cell expresses the antibody.

21. The engineered NK cell of claim 1, wherein the cytokine is membrane-bound.

22. The engineered NK cell of claim 1, wherein the cytokine is IL-15.

23. The engineered NK cell of claim 1, wherein the NK cell is further modified to express one or more additional heterologous proteins selected from the group consisting of an antigen receptor, a cytokine, a homing receptor, a chemokine receptor, and a combination thereof.

24. The engineered NK cell of claim 1, wherein the NK cells are pre-activated with one or more cytokines.

25. The engineered NK cell of claim 24, wherein the cytokines are IL-2, IL-7, IL-12, IL-15, IL- 18, IL-21, or a combination thereof.

26. The engineered NK cell of claim 1, wherein the NK cell further comprises one or more engineered mutations in an endogenous gene.

27. The engineered NK cell of claim 26, wherein the endogenous gene is TGFBR2, CISH, and/or CD38.

28. A composition comprising the engineered NK cell of any one of claims 1 to 27.

29. A composition comprising a complex comprising: a) an engineered NK cell modified to express part or all of the CD3 receptor complex, an iTCRa chain, an iTCRP, and one or more cytokines; and b) a bispecific or multi-specific antibody, wherein the bispecific or multi-specific antibody comprises an anti-CD3 antibody that is bound to CD3 on the NK cells.

30. The composition of claim 29, wherein the engineered NK is cell modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT- NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122).

31. The composition of claim 29, wherein the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCRP chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315.

32. The composition of claim 29, wherein the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCRP chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 301, 307, 311, or 313.

33. The composition of claim 29, wherein the bispecific or multi-specific antibody is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B.

34. The composition of claim 29, wherein the bispecific or multi-specific antibody is Blinatumomab.

35. The composition of claim 29, further comprising a pharmaceutically acceptable excipient.

36. The composition of claim 29, wherein the composition is comprised in a delivery device.

37. A method of treating a disease in an individual, the method comprising the step of administering to the individual a therapeutically effective amount of any one of the cells or compositions of claims 1 to 27 or 29 to 36.

38. The method of claim 37, wherein the disease is an autoimmune disease, infection, and/or cancer.

39. A method of treating a disease in an individual, the method comprising the step of administering to the individual an engineered NK cell modified to express a) part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3 b) part or all of a single chain or any combination of invariant NK T cell receptor (iTCR) alpha (iTCRa) chain or beta (iTCR0) chain; and c) a cytokine selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, IL-7, GMCSF, and a combination thereof.

40. The method of claim 39, further comprising administering to the individual at the same time or at different time, one or more bispecific or multispecific antibodies.

41. The method of claim 40, wherein the one or more bispecific or multispecific antibodies is Blinatumomab, Tebentafusp, Mosunetuzumab, Teclistamab, Glofitamab, Epcoritamab, Flotetuzumab, APV0436, and/or TNB383B.

42. The method of claim 40, wherein the one or more bispecific or multispecific antibodies and the engineered NK cell are administered at the same time, and/or wherein the one or more bispecific or multispecific antibodies and the engineered NK cells are complexed prior to administration to the individual.

43. The method of claim 39, wherein the engineered NK is cell modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 118), UT- NK15-28 (SEQ ID NO: 120), or UTNK15-28-DAP10 (SEQ ID NO: 122).

44. The method of claim 39, wherein the engineered NK cell is modified to express an iTCRa chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 293, and an iTCRP chain comprising an amino acid sequence at least 90% identical to SEQ ID NOs: 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, or 315.

45. The method of any one of claims 39 to 44, wherein the disease is cancer.