CD3-expressing natural killer cells with enhanced function for adoptive immunotherapy

Abstract

Embodiments of the present disclosure include methods and compositions for modifying NK cells by a human hand to express a T cell receptor and a CD3 coreceptor on NK cells that are not naturally expressed. Such modified NK cells act effectively with a monospecific antibody, a bispecific antibody, or a multispecific antibody, and the bispecific antibody or multispecific antibody is adjusted to consist of an anti-CD3 antibody that binds to the modified NK cells, thereby inducing signal transduction, activation, and cytotoxicity of target cells to which the antibody also binds. Thus, NK cells are specially configured to be able to act effectively with bispecific NK cell engagers (BiKEs) and bispecific T cell engagers (BiTEs).

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to International Application No. PCT / US2022 / 074062, filed on July 22, 2022, and U.S. Provisional Patent Application No. 63 / 481,588, filed on January 25, 2023, each of which is hereby incorporated by reference in its entirety.

[0002] Sequence Listing This application includes a sequence listing submitted in ST26 format, which is hereby incorporated by reference in its entirety. The ST26 copy created on July 21, 2023, is named MDAC_P1347WO_Sequence_Listing.xml and is 422,889 bytes in size.

[0003] Technical Field The present disclosure relates to the fields of at least immunology, cell biology, molecular biology, and medicine (including at least cancer medicine).

[0004] Natural killer (NK) cells have been studied as potential anti - tumor effectors, but many barriers, mainly related to the lack of antigen specificity, limit their therapeutic use. One approach to overcome this is to transduce NK cells with chimeric antigen receptors (CARs) or engineered T - cell receptors (TCRs) to target desired antigens. In T cells, bispecific antibodies or multispecific antibodies such as bispecific T - cell engagers (BiTEs) that bind to CD3 on the T - cell surface and also to antigens on the cancer cell surface can be utilized. CD3 is composed of four different chains, and in mammals, the complex includes the CD3γ chain, the CD3δ chain, and two CD3ε chains. These chains bind to the T - cell receptor (TCR) and the ζ chain (zeta chain) and generate activation signals in T lymphocytes. However, NK cells do not naturally express the CD3 receptor complex or TCR.

Summary of the Invention

Problems to be Solved by the Invention

[0005] The present disclosure meets a long - standing need in the art of improving immunotherapies, including immunotherapies that utilize NK cells.

Means for Solving the Problems

[0006] Embodiments of the present disclosure include methods and compositions for treating an individual having cancer using adoptive cell therapy. In a specific embodiment, the individual is provided with a therapeutically effective amount of a two - part therapy that includes both modified NK cells and an antibody capable of binding to the NK cells to initiate signal transduction, activation, and killing of target cells. The present disclosure relates to NK cells modified to express a plurality of proteins that do not naturally occur in NK cells but function in concert, including heterologous proteins on the surface of NK cells that are not naturally present.

[0007] In a specific embodiment, the NK cells are engineered to express one or more proteins from the CD3 coreceptor complex and optionally the TCR receptor complex, each of which is normally present on the T - cell surface. Such engineering improves the versatility with which NK cells can be utilized with various bispecific or multispecific antibodies, including anti - CD3 antibodies (e.g., anti - CD3 scFv). In certain embodiments, the modified NK cells are administered in combination with one or more monospecific, bispecific, or multispecific antibodies to an individual in need thereof, and each bispecific or multispecific antibody has one antibody that targets CD3 and one antibody that binds to a desired antigen such as a cancer antigen. As a result, in specific cases, NK cells expressing CD3 can bind to the anti - CD3 antibody portion of the bispecific or multispecific antibody, and the antibody that binds to the cancer antigen binds to the cancer antigen on the surface of cancer cells. Such cooperative binding of the NK cells and the antibody results in the activation of cytotoxic activity against the target cancer antigen.

[0008] In certain embodiments, the disclosure relates to modified NK cells that express a full or partial CD3 complex with or without a TCR, where in some cases, individual CD3 chain(s) are heterologously linked to an NK-associated signaling domain, all of which enable the use of the modified NK cells with various bispecific antibodies.

[0009] Embodiments of the present disclosure include compositions comprising NK cells modified to express a single chain or any combination of CD3δ, CD3ε, CD3γ, or CD3ζ, in whole or in part. Optionally, the NK cells are modified to express the T cell receptor (TCR) αβ chain or TCRγδ chain. The NK cells can be modified to express CD3ζ, two of CD3ε, CD3δ, and a part or all of CD3γ. Optionally, the NK cells are modified to express the full length of CD3ζ, CD3ε, CD3δ, and / or CD3γ. In particular, any one or more of CD3ζ, CD3ε, CD3δ, and CD3γ are heterologously linked to one or more intracellular signaling domains. The intracellular signaling domain can be selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and combinations thereof. In some embodiments, the intracellular signaling domain is fused to CD3ζ. In some embodiments, the intracellular signaling domain is derived from DAP10. In some embodiments, the intracellular signaling domain is derived from CD28. In some embodiments, the intracellular signaling domain comprises a sequence derived from DAP10 and a sequence derived from CD28. In certain embodiments, the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 115. In certain embodiments, the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 116. In certain embodiments, the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 117. In certain embodiments, the NK cells are modified to express a polynucleotide sequence that is 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 intracellular signaling domain can also include other co-stimulatory signals associated with NK cell function, including but not limited to 2B4, DNA, 4-1BB, DAP12, NKG2D, etc.In specific embodiments, the composition further comprises one or more monospecific antibodies, bispecific antibodies, or multispecific antibodies, and the bispecific or multispecific antibodies comprise anti-CD3 antibodies. NK cells can express antibodies and / or be complexed with antibodies. In some embodiments, the antibodies are ipilimumab, amivantamab, and / or cetuximab. In some embodiments, the TCR is directed to a cancer antigen or a viral antigen. In specific embodiments, the NK cells are derived from umbilical cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or mixtures thereof. In some embodiments, the TCR is directed to the KRAS antigen. NK cells can be pre-activated with, for example, one or more cytokines including IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, or combinations thereof. In some embodiments, the NK cells are expanded in the presence of IL-2. In specific embodiments, the NK cells are modified to express one or more heterologous proteins, such as one or more engineered antigen receptors, one or more cytokines, one or more homing receptors, and / or one or more chemokine receptors. In specific cases, the engineered antigen receptor is a chimeric antigen receptor and / or an engineered T cell receptor. In some cases, the heterologous protein is a cytokine, for example, selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, GMCSF, or combinations thereof. The cytokine can be membrane-bound, and the membrane-bound cytokine can comprise a transmembrane domain from CD8, CD28, CD27, B7H3, IgG1, IgG4, CD4, DAP10, or DAP12. In specific cases, the NK cells express a chimeric antigen receptor and a cytokine. In some cases, the bispecific antibody comprises an antibody that targets a cancer antigen. In some cases, the bispecific antibody comprises amivantamab. In some cases, the antibody comprises ipilimumab and / or cetuximab. In some cases, the antibody targets EGFR, C-met, and / or TROP-2.

[0010] Embodiments of the present disclosure include a composition comprising: (1) NK cells modified to express some or all of the CD3 receptor complex and optionally modified to express the T cell receptor (TCR) αβ chain or TCRγδ chain; and (2) a single - specificity, bispecific or multispecific antibody, which is a bispecific or multispecific antibody comprising an anti - CD3 antibody that binds to CD3 on the NK cells. In a specific embodiment, the complex is contained in a pharmaceutically acceptable excipient. The complex can be contained in a delivery device.

[0011] In certain embodiments, a method of treating cancer in an individual is provided, comprising administering to the individual a therapeutically effective amount of any one of the compositions included herein. In some embodiments, the NK cells and the antibody are administered to the individual simultaneously. The NK cells and the antibody can or cannot be administered in the same formulation. The NK cells and the antibody can be pre - complexed before administration to the individual. In a specific embodiment, the NK cells and the antibody are administered to the individual at different times. The NK cells and the antibody can be administered by infusion. In a specific embodiment, the NK cells are autologous or allogeneic with respect to the individual.

[0012] Embodiments of the present disclosure include a method of redirecting the specificity of NK cells against a cancer antigen for treating an individual with a single - specificity, bispecific or multispecific antibody, wherein the bispecific or multispecific antibody comprises an anti - CD3 antibody, and the method comprises administering to the individual the antibody and NK cells that express some or all of the CD3 receptor complex and optionally some or all of the TCRαβ chain or TCRγδ chain. In a specific embodiment, the method further comprises the step of modifying the NK cells to express some or all of the CD3 receptor complex. In a specific embodiment, the method further comprises the step of modifying the NK cells to express the TCRαβ chain or TCRγδ chain. Optionally, the method further comprises the step of modifying the NK cells to express one or more heterologous proteins.

[0013] Certain specific embodiments of the present invention are characterized through the following aspects.

[0014] Aspect 1 is a composition comprising engineered NK cells modified to express one or more transgenic polynucleotides encoding at least one engineered TCR, wherein the engineered TCR targets a KRAS antigen, and comprises: a) a CD3 protein complex comprising a single chain or any combination of a part or all of CD3δ, CD3ε, CD3γ, or CD3ζ; b) optionally, at least one cytokine; and c) one or more of TCRα chain, TCRβ chain, TCRγ chain, and / or TCRδ chain.

[0015] Aspect 2 is the composition according to aspect 1, wherein the NK cells are modified to express a TCRαβ chain or a TCRγδ chain.

[0016] Aspect 3 is the composition according to aspect 1 or 2, wherein the NK cells are modified to express CD3ζ, two of CD3ε, CD3δ, and a part or all of CD3γ.

[0017] Aspect 4 is the composition according to any one of aspects 1 to 3, wherein the NK cells are modified to express the full length of CD3ζ, CD3ε, CD3δ, and / or CD3γ.

[0018] Aspect 5 is the composition according to any one of aspects 1 to 4, wherein any one or more of CD3ζ, CD3ε, CD3δ, and CD3γ are heterologously linked to one or more intracellular signaling domains.

[0019] Aspect 6 is the composition according to aspect 5, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and combinations thereof.

[0020] Aspect 7 is the composition according to aspect 5 or 6, wherein the intracellular signaling domain comprises a DAP10 intracellular signaling domain.

[0021] Aspect 8 is the composition according to aspect 7, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 115.

[0022] Aspect 9 is the composition according to aspect 7, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 115.

[0023] Aspect 10 is the composition according to any one of aspects 6 to 9, wherein the intracellular signaling domain comprises the CD28 intracellular signaling domain.

[0024] Aspect 11 is the composition according to aspect 10, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 116.

[0025] Aspect 12 is the composition according to aspect 10, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 116.

[0026] Aspect 13 is the composition according to any one of aspects 6 to 13, wherein the intracellular signaling domain comprises the DAP10 and CD28 intracellular signaling domains.

[0027] Aspect 14 is the composition according to aspect 13, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 117.

[0028] Aspect 15 is the composition according to aspect 13, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 117.

[0029] Aspect 16 is the composition according to any one of aspects 1 to 15, wherein the composition further comprises one or more monospecific antibodies, bispecific antibodies, or multispecific antibodies, and the bispecific antibody or multispecific antibody comprises an anti-CD3 antibody.

[0030] Aspect 17 is the composition according to aspect 16, wherein the NK cells express the antibody.

[0031] Aspect 18 is the composition according to aspect 16 or 17, wherein the NK cells are complexed with an antibody.

[0032] Aspect 19 is the composition according to any one of aspects 16 to 18, wherein the antibody is ipilimumab, amivantamab, and / or cetuximab.

[0033] Aspect 20 is the composition according to any one of aspects 1 to 19, wherein the NK cells are derived from umbilical cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or a mixture thereof.

[0034] Aspect 21 is the composition according to any one of aspects 1 to 20, wherein the NK cells are pre-activated.

[0035] Aspect 22 is the composition according to aspect 21, wherein the NK cells are pre-activated with one or more cytokines.

[0036] Aspect 23 is the composition according to aspect 22, wherein the cytokine is IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, or a combination thereof.

[0037] Aspect 24 is the composition according to any one of aspects 1 to 23, wherein the NK cells are expanded.

[0038] Aspect 25 is the composition according to aspect 24, wherein the NK cells are expanded in the presence of IL-2.

[0039] Aspect 26 is the composition according to any one of aspects 1 to 25, wherein the NK cells are modified to express one or more additional heterologous proteins.

[0040] Aspect 27 is the composition according to aspect 26, wherein the additional heterologous protein is an engineered antigen receptor, cytokine, homing receptor, and / or chemokine receptor.

[0041] Aspect 28 is the composition according to any one of Aspects 1 to 27, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 299 and a sequence that is at least 85% identical to SEQ ID NO: 301.

[0042] Aspect 29 is the composition according to any one of Aspects 1 to 28, wherein the TCR comprises one or more sequences that are at least 85% identical to SEQ ID NOs: 299 to 317.

[0043] Aspect 30 is the composition according to any one of Aspects 1 to 29, wherein the target KRAS antigen epitope comprises or consists of GADGVGKSA (SEQ ID NO: 293) and / or GADGVGKSAL (SEQ ID NO: 292).

[0044] Aspect 31 is the composition according to any one of Aspects 1 to 30, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 304 and a sequence that is at least 85% identical to SEQ ID NO: 305.

[0045] Aspect 32 is the composition according to any one of Aspects 1 to 31, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 306 and a sequence that is at least 85% identical to SEQ ID NO: 307.

[0046] Aspect 33 is the composition according to any one of Aspects 1 to 32, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 308 and a sequence that is at least 85% identical to SEQ ID NO: 309.

[0047] Aspect 34 is the composition according to any one of Aspects 26 to 33, wherein the heterologous protein is a cytokine.

[0048] Aspect 35 is the composition according to Aspect 34, wherein the cytokine is selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, IL-7, GMCSF, or a combination thereof.

[0049] Aspect 36 is the composition according to Aspect 34 or 35, wherein the cytokine is membrane-bound.

[0050] Aspect 37 is the composition according to any one of Aspects 34 to 36, wherein the cytokine is IL-15.

[0051] Aspect 38 is the composition according to Aspect 36 or 37, wherein the membrane-bound cytokine comprises a transmembrane domain of CD8, CD28, CD27, B7H3, IgG1, IgG4, CD4, DAP10, or DAP12.

[0052] Aspect 39 is the composition according to any one of Aspects 27 to 38, wherein the NK cells express a chimeric antigen receptor and a cytokine.

[0053] Aspect 40 is the composition according to any one of Aspects 16 to 39, wherein the monospecific antibody, bispecific antibody, or multispecific antibody comprises an antibody that targets a cancer antigen.

[0054] Aspect 41 is the composition according to Aspect 40, wherein the cancer antigen is an EGFR, C-met, and / or TROP-2 antigen.

[0055] Aspect 42 is a composition comprising a complex comprising a) NK cells modified to express part or all of the CD3 receptor complex and modified to express a T cell receptor (TCR) αβ chain or TCRγδ chain; and b) a monospecific antibody, bispecific antibody, or multispecific antibody comprising an anti-CD3 antibody that binds to CD3 on the NK cells, wherein the TCR targets a KRAS antigen.

[0056] Aspect 43 is the composition according to Aspect 42, wherein the NK cells are modified to express a TCRαβ chain that is at least 85% identical to SEQ ID NO: 299 and SEQ ID NO: 301, and the antibody is ipilimumab, amivantamab, and / or cetuximab.

[0057] Aspect 44 is the composition according to aspect 42 or 43, wherein the NK cells are modified to express full-length CD3ζ, CD3ε, CD3δ, and / or CD3γ.

[0058] Aspect 45 is the composition according to any one of aspects 42 to 44, wherein any one or more of CD3ζ, CD3ε, CD3δ, and CD3γ are heterologously linked to one or more intracellular signaling domains.

[0059] Aspect 46 is the composition according to aspect 45, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, DNAM, and combinations thereof.

[0060] Aspect 47 is the composition according to aspect 45 or 46, wherein the intracellular signaling domain comprises the DAP10 intracellular signaling domain.

[0061] Aspect 48 is the composition according to any one of aspects 45 to 47, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 115.

[0062] Aspect 49 is the composition according to any one of aspects 45 to 48, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 115.

[0063] Aspect 50 is the composition according to any one of aspects 45 to 49, wherein the intracellular signaling domain comprises the CD28 intracellular signaling domain.

[0064] Aspect 51 is the composition according to any one of aspects 45 to 50, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 116.

[0065] Aspect 52 is the composition according to any one of Aspects 45 to 51, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 116.

[0066] Aspect 53 is the composition according to any one of Aspects 45 to 52, wherein the intracellular signaling domain comprises the DAP10 and CD28 intracellular signaling domains.

[0067] Aspect 54 is the composition according to any one of Aspects 45 to 53, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 117.

[0068] Aspect 55 is the composition according to any one of Aspects 45 to 54, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 117.

[0069] Aspect 56 is the composition according to any one of Aspects 1 to 55, wherein the composition is contained in a pharmaceutically acceptable excipient.

[0070] Aspect 57 is the composition according to any one of Aspects 1 to 56, wherein the composition is contained in a delivery device.

[0071] Aspect 58 is a method for treating cancer in an individual, comprising the step of administering to the individual a therapeutically effective amount of any one of the compositions according to Aspects 1 to 57.

[0072] Aspect 59 is a method for treating cancer in an individual, comprising the step of administering to the individual a therapeutically effective amount of any one of the compositions according to Aspects 16 to 57, wherein the NK cells and the antibody are administered to the individual simultaneously, and optionally, the NK cells and the antibody are administered in the same formulation and / or pre-complexed before being administered to the individual.

[0073] Aspect 60 is the method according to Aspect 58, wherein the NK cells and the antibody are administered to the individual at different times.

[0074] Aspect 61 is the method according to any one of Aspects 58 to 60, wherein NK cells and an antibody are administered by injection.

[0075] Aspect 62 is the method according to any one of Aspects 58 to 61, wherein the NK cells are autologous with respect to the individual.

[0076] Aspect 63 is the method according to any one of Aspects 58 to 61, wherein the NK cells are allogeneic with respect to the individual.

[0077] Aspect 64 is a method of redirecting the specificity of NK cells against a cancer antigen for the treatment of an individual with a monospecific antibody, bispecific antibody or multispecific antibody, wherein the bispecific antibody or multispecific antibody comprises an anti-CD3 antibody, and administering to the individual the antibody and NK cells that express a part or all of the CD3 receptor complex and express a part or all of the TCRαβ chain or TCRγδ chain, and the TCR targets the KRAS antigen.

[0078] Aspect 65 is the method according to Aspect 64, wherein the NK cells are modified to express full-length CD3ζ, CD3ε, CD3δ, and / or CD3γ.

[0079] Aspect 66 is the method according to Aspect 64 or 65, wherein any one or more of D3ζ, CD3ε, CD3δ, and CD3γ are heterologously linked to one or more intracellular signaling domains.

[0080] Aspect 67 is the method according to any one of Aspects 64 to 66, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, DNAM, and combinations thereof.

[0081] Aspect 68 is the method according to Aspect 66 or 67, wherein the intracellular signaling domain comprises the DAP10 intracellular signaling domain.

[0082] Aspect 69 is the method according to any one of Aspects 66 to 68, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 115.

[0083] Aspect 70 is the method according to any one of Aspects 66 to 69, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 115.

[0084] Aspect 71 is the method according to any one of Aspects 66 to 70, wherein the intracellular signaling domain comprises the CD28 intracellular signaling domain.

[0085] Aspect 72 is the method according to any one of Aspects 66 to 71, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 116.

[0086] Aspect 73 is the method according to any one of Aspects 66 to 72, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 116.

[0087] Aspect 74 is the method according to any one of Aspects 66 to 73, wherein the intracellular signaling domain comprises the DAP10 and CD28 intracellular signaling domains.

[0088] Aspect 75 is the method according to any one of Aspects 66 to 74, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO: 117.

[0089] Aspect 76 is the method according to any one of Aspects 66 to 75, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO: 117.

[0090] Aspect 77 is the method according to any one of Aspects 64 to 76, wherein the TCR chain is a TCRαβ chain and is at least 85% identical to SEQ ID NO: 299 and SEQ ID NO: 301.

[0091] Aspect 78 is the method according to any one of Aspects 64 to 77, wherein the TCR chain is a TCRαβ chain and is at least 85% identical to one or more of SEQ ID NOs: 299 to 317.

[0092] Aspect 79 is the method according to any one of Aspects 64 to 78, wherein the target KRAS antigen epitope comprises and / or consists of GADGVGKSA (SEQ ID NO: 293) and / or GADGVGKSAL (SEQ ID NO: 292).

[0093] Aspect 80 is the method according to any one of Aspects 64 to 79, wherein the TCR chain comprises a sequence that is at least 85% identical to SEQ ID NO: 304 and a sequence that is at least 85% identical to SEQ ID NO: 305.

[0094] Aspect 81 is the method according to any one of Aspects 64 to 79, wherein the TCR chain comprises a sequence that is at least 85% identical to SEQ ID NO: 306 and a sequence that is at least 85% identical to SEQ ID NO: 307.

[0095] Aspect 82 is the method according to any one of Aspects 64 to 79, wherein the TCR chain comprises a sequence that is at least 85% identical to SEQ ID NO: 308 and a sequence that is at least 85% identical to SEQ ID NO: 309.

[0096] Aspect 83 is the method according to any one of Aspects 64 to 82, further comprising the step of modifying NK cells to express one or more additional heterologous proteins.

[0097] Other objects, features, and advantages of the present invention will become apparent from the following detailed description. However, it should be understood that the detailed description and specific examples, while indicating specific embodiments of the present invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art from this detailed description.

[0098] The following drawings form a part of this specification and are included to further illustrate certain aspects of the present invention. The present invention can be better understood by reference to one or more of these drawings in combination with the detailed description of the specific embodiments presented herein.

Brief Description of the Drawings

[0099]

Figure 1-1

Figure 1-2

Figure 1-3

[0100]

Figure 2-1

Figure 2-2

[0101]

Figure 3

[0102]

Figure 4

[0103]

Figure 5

[0104]

Figure 6

[0105]

Figure 7

[0106]

Figure 8

[0107]

Figure 9

[0108]

Figure 10-1

Figure 10-2

[0109]

Figure 11

[0110]

Figure 12

[0111]

Figure 13

[0112]

Figure 14

[0113]

Figure 15

[0114]

Figure 16

[0115]

Figure 17

[0116]

Figure 18

[0117]

Figure 19

[0118]

Figure 20

[0119]

Figure 21

[0120]

Figure 22-1

Figure 22-2

Figure 22-3

[0121]

Figure 23-1

Figure 23-2

[0122]

Figure 24-1

Figure 24-2

Figure 24-3

Figure 24-4

[0123]

Figure 25

[0124]

Figure 26-1

Figure 26-2

Figure 26-3

[0125]

Figure 27-1

Figure 27-2

Mode for Carrying Out the Invention

[0126] Detailed Description In accordance with long-standing patent law practice, as used herein, when the words "a" and "an" are used in combination with the word "comprising", including in the claims, they represent "one or more". Some embodiments of the present disclosure may consist of, or consist essentially of, one or more elements, method steps, and / or methods of the present disclosure. It is contemplated that any method or composition described herein may be practiced with respect to any other method or composition described herein, and that different embodiments may be combined.

[0127] Throughout this specification, unless the context requires otherwise, the words "comprise", "comprises", and "comprising" are understood to mean including the stated step or element or group of steps or elements but not to mean excluding other steps or elements or group of steps or elements. "Consisting of" means including and limited to what follows the phrase "consisting of". Thus, the expression "consisting of" indicates that the recited elements are essential or mandatory and that no other elements may be present. "Consisting essentially of" means including the elements recited after that phrase and limited to other elements that do not interfere with or contribute to the activity or action specified for the recited elements in the disclosure. Thus, the expression "consisting essentially of" indicates that the recited elements are essential or mandatory but that other elements are optional and may or may not be present depending on whether they affect the activity or action of the recited elements.

[0128] Throughout this specification, references to "one embodiment", "an embodiment", "a particular embodiment", "related embodiments", "an embodiment", "additional embodiments", or "further embodiments", or combinations thereof, mean that the particular features, structures, or characteristics described in connection with the embodiment are included in at least one embodiment of the invention. Thus, although the foregoing phrases appear in various places throughout this specification, they 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.

[0129] As used herein, the terms "or" and "and / or" are used to combine multiple components or to describe them mutually exclusively. 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 the embodiments.

[0130] Throughout this application, the term "about" is used in accordance with its plain and ordinary meaning in the fields of cell biology and molecular biology, indicating that a value includes the standard deviation of error for the apparatus or method employed to determine that value.

[0131] As used herein, the term "CD3 receptor complex" or "CD3 coreceptor complex" refers to a protein complex that naturally functions as a T cell coreceptor and is composed of the CD3ζ chain, CD3γ chain, CD3δ chain, and two CD3ε chains (although alternatively only one CD3ε chain is used).

[0132] As used herein, the term "engineered" refers to an entity generated by a human hand, including cells, nucleic acids, polypeptides, vectors, etc. In at least some cases, the engineered entity is synthetic and contains elements that do not exist in nature or are not constructed by the methods utilized in the present disclosure. In a specific embodiment, a vector is engineered by recombinant nucleic acid technology, and a cell is engineered by transfection or transduction with the engineered vector. In a specific embodiment, the engineered cell is a cell comprising modification of one or more exogenous antigen receptors (e.g., chimeric antigen receptors, T cell receptors, etc.), suicide genes, cytokines and / or cytokine receptors (e.g., IL-15, IL-15R, etc.), chemokine / homing receptors, and / or one or more endogenous genes.

[0133] The phrase "pharmaceutically or pharmacologically acceptable" appropriately refers to molecular entities and compositions that do not produce adverse reactions, allergic reactions, or other adverse reactions when administered to animals such as humans. The preparation of pharmaceutical compositions containing antibodies or additional active ingredients will be known to those skilled in the art in light of the present disclosure. Further, it will be understood that when administered to an animal (e.g., a human), the preparation should meet the standards of sterility, pyrogenicity, general safety, and purity required by the FDA's Office of Biological Standards.

[0134] As used herein, "pharmaceutically acceptable carrier" includes any and all aqueous solvents (e.g., water, alcohol / aqueous solutions, physiological saline, parenteral vehicles such as sodium chloride, Ringer's dextrose), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters such as ethyl oleate), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, antioxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, excipients, disintegrants, lubricants, sweetening agents, flavoring agents, dyes, body fluids, and nutrient supplements, and such materials and combinations thereof will be known to those 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.

[0135] As used herein, the term "subject" generally refers to an individual having or suspected of having cancer. The subject can be any organism or animal subject to a method or material, including mammals such as 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, for example, a patient having or suspected of having a benign or malignant neoplasm or a disease such as cancer (sometimes referred to as a medical condition). The subject can be undergoing treatment or have been treated. The subject may be asymptomatic. The subject may be a healthy individual but may wish to prevent cancer. The term "individual" is used interchangeably at least in some cases. As used herein, a "subject" or "individual" may or may not be housed in a medical facility and may be treated as an outpatient of a medical facility. An individual may receive "one or more pharmaceutical compositions" via the Internet. An individual may include any age of human or non-human animal and thus includes both adults and juveniles (i.e., children) and infants, including individuals in utero. Since this term does not imply a need for medical treatment, an individual can participate in an experiment, whether clinical or in support of basic science research, whether voluntary or involuntary.

[0136] As used herein, "treatment" or "therapy" includes any beneficial or desirable effect on the symptoms or pathology of a disease or pathological condition, including even a minimal decrease in one or more measurable markers of the disease or condition being treated, such as cancer. Treatment can optionally include either a reduction or improvement in one or more symptoms of the disease or condition, or a delay in the progression of the disease or condition. "Treatment" does not necessarily indicate complete eradication or cure of the disease or condition or the symptoms associated therewith.

[0137] As used herein, the "TCR / CD3 complex" refers to a protein complex that is naturally found on the surface of T cells and includes, in addition to the CD3ζ, CD3γ, CD3δ, and CD3ε chains, the T cell receptor (TCR) α and β chains, and / or the T cell receptor γ and δ chains.

[0138] I. Embodiments of the Present Disclosure Natural killer (NK) cells are a new form of cellular immunotherapy for patients with malignant blood diseases, as well as for solid tumors. The present disclosure specifically relates to NK cells that are modified to have enhanced functions as immunotherapy compared to unmodified NK cells. This modification enables NK cells to have higher versatility when used together with other therapeutic agents and, in at least 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 either (i) a single CD3 chain (CD3 zeta, CD3 epsilon, CD3 delta, or CD3 gamma), or a part or all of the human CD3 receptor complex (including any combination of CD3 delta, epsilon (1 or 2 copies of epsilon), gamma, and zeta), or (ii) a single CD3 chain or the human CD3 receptor complex (including any combination of CD3 delta, epsilon (1 or 2 molecules), gamma, and zeta) as a full-length protein or partial protein heterologously linked to one or more intracellular signaling domains; (iii) the CD3 complex may or may not include a T cell receptor (αβ or γδ). The present disclosure relates to the use of CD3-expressing NK cells in the diagnosis and treatment of diseases, including the use of cells combined with a monospecific, bispecific, or multispecific antibody, where one epitope of the bispecific antibody and / or multispecific antibody binds to CD3 on the CD3-expressing NK cells. The CD3-expressing NK cells can be pre-complexed with the bispecific / multispecific antibody ex vivo and / or combined in vivo to redirect their specificity for the target antigen. In diagnostic embodiments, the labeled NK cells can be loaded with any type of bispecific or multispecific antibody containing at least an anti-CD3 antibody, and the loaded labeled NK cells can be monitored for transport to the site of the target antigen to which another antibody on the bispecific or multispecific antibody binds.

[0139] In certain embodiments of the present disclosure, the TCR recognizes an antigen and / or epitope presented by a major histocompatibility complex (MHC). In certain embodiments, the antigen and / or epitope is a peptide, lipid, and / or glycolipid. In certain embodiments, the MHC is class I MHC. In certain embodiments, the MHC is class II MHC. In certain embodiments, the MHC is a non-classical MHC.

[0140] In certain embodiments, the TCR target antigen can be a primary antigen and / or a secondary antigen that provides target antigen specificity to the transduced effector cell. In certain embodiments, the TCR acts primarily as a stabilizer of the CD3 coreceptor complex, while the antibody provides primary target antigen specificity to the transduced effector cell.

[0141] II. Compositions of the Present Disclosure The present disclosure relates to a composition comprising at least modified NK cells that express at least a portion of the TCR / CD3 complex. Optionally, the composition may also include a monospecific, bispecific, or multispecific antibody, including in the same formulation. In alternative embodiments, however, the NK cells and the antibody are utilized as physically separate compositions.

[0142] A. TCR / CD3 Modification of NK Cells In certain embodiments, provided herein are compositions comprising NK cells manually modified to express some or all of the TCR receptor complex and some or all of the CD3 coreceptor complex. In specific embodiments, the NK cells are modified to include all components of the CD3 complex, including CD3ζ, CD3ε, CD3γ, and CD3δ. In certain cases, the full lengths of CD3ζ, CD3ε, CD3γ, and CD3δ are utilized, including their extracellular, transmembrane, and intracellular domains, although in alternative embodiments, only a portion of one or more of CD3ζ, CD3ε, CD3γ, and CD3δ are utilized, each of which may or may not be combined with one or more intracellular signaling domains such as CD16, NKG2D, DAP10, DAP12, CD28, 41BB, 2B4, CD27, OX40, or any combination thereof. The NK cells may also be modified to express the TCR receptor complex, although in alternative embodiments, none of the TCR receptor complex components are utilized.

[0143] In certain embodiments, an amino acid sequence (e.g., a polypeptide) may include an amino acid represented by the single letter "X" or the three letter code "Xaa". In some embodiments, the amino acid represented by "X" or "Xaa" is any naturally occurring amino acid, e.g., 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 (Gln, Q), glycine (Gly, G), proline (Pro, P), cysteine (Cys, C), alanine (Ala, A), valine (Val, V), isoleucine (Ile, I), leucine (Leu, L), methionine (Met, M), phenylalanine (Phe, F), tyrosine (Tyr, Y), or tryptophan (Trp, W).

[0144] 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 (Gln, 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 (Val, V). In some embodiments, the amino acid represented by "X" or "Xaa" in SEQ ID NO: 25 or SEQ ID NO: 88 is isoleucine (Ile, 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 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).

[0145] In certain embodiments, any specific sequence of CD3 receptor components, including wild-type or mutant forms of the components, is utilized as long as the CD3 receptor having the mutant enables signal transduction via the CD3 complex leading to activation and killing of the target. Optionally, the following sequence examples of CD3ε, CD3δ, CD3γ, and CD3ζ are utilized for modification of NK cells.

[0146] CD3 epsilon (UniProtKB - P07766 (CD3E_HUMAN))

[0147] Signal peptide MQSGTHWRVLGLCLLSVGVW (SEQ ID NO: 1)

[0148] Extracellular domain sp|P07766|23-126 DGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD (SEQ ID NO: 2)

[0149] Transmembrane domain sp|P07766|127-152 VMSVATIVIVDICITGGLLLLVYYWS (SEQ ID NO: 3)

[0150] Intracellular domain sp|P07766|153-207 KNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI (SEQ ID NO: 4)

[0151] An example of the Homo sapiens CD3e molecule (CD3E), mRNA is in the NCBI reference sequence: GENBANK® accession number NM_000733.4 ATGCAGTCGGGCACTCACTGGAGAGTTCTGGGCCTCTGCCTCTTATCAGTTGGCGTTTGGGGGCAAGATGGTAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCTCTGGAACCACAGTAATATTGACATGCCCTCAGTATCCTGGATCTGAAATACTATGGCAACACAATGATAAAAACATAGGCGGTGATGAGGATGATAAAAACATAGGCAGTGATGAGGATCACCTGTCACTGAAGGAATTTTCAGAATTGGAGCAAAGTGGTTATTATGTCTGCTACCCCAGAGGAAGCAAACCAGAAGATGCGAACTTTTATCTCTACCTGAGGGCAAGAGTGTGTGAGAACTGCATGGAGATGGATGTGATGTCGGTGGCCACAATTGTCATAGTGGACATCTGCATCACTGGGGGCTTGCTGCTGCTGGTTTACTACTGGAGCAAGAATAGAAAGGCCAAGGCCAAGCCTGTGACACGAGGAGCGGGTGCTGGCGGCAGGCAAAGGGGACAAAACAAGGAGAGGCCACCACCTGTTCCCAACCCAGACTATGAGCCCATCCGGAAAGGCCAGCGGGACCTGTATTCTGGCCTGAATCAGAGACGCATCTGA (SEQ ID NO: 5)

[0152] Examples showing the entire CD3 epsilon sequences of each nucleic acid and amino acid are shown below (the underlines indicate signal peptide sequences): ATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGGGCCAG GACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAATC (SEQ ID NO: 37) MQSGTHWRVLGLCLLSVGVWGQ DGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI (SEQ ID NO: 38)

[0153] CD3 delta (UniProtKB - P04234 (CD3D_HUMAN))

[0154] Signal peptide MEHSTFLSGLVLATLLSQVS (SEQ ID NO: 6)

[0155] Extracellular domain sp|P04234|22-105 FKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVA (SEQ ID NO: 7)

[0156] Transmembrane domain sp|P04234|106-126 GIIVTDVIATLLLALGVFCFA (SEQ ID NO: 8)

[0157] Intracellular domain sp|P04234|127-171 GHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK (SEQ ID NO: 9)

[0158] Homo sapiens CD3d molecule, delta (CD3-TCR complex), mRNA (cDNA clone MGC:88324 IMAGE:30412345), complete cds GENBANK(R):BC070321.1 ATGGAACATAGCACGTTTCTCTCTGGCCTGGTACTGGCTACCCTTCTCTCGCAAGTGAGCCCCTTCAAGATACCTATAGAGGAACTTGAGGACAGAGTGTTTGTGAATTGCAATACCAGCATCACATGGGTAGAGGGAACGGTGGGAACACTGCTCTCAGACATTACAAGACTGGACCTGGGAAAACGCATCCTGGACCCACGAGGAATATATAGGTGTAATGGGACAGATATATACAAGGACAAAGAATCTACCGTGCAAGTTCATTATCGAATGTGCCAGAGCTGTGTGGAGCTGGATCCAGCCACCGTGGCTGGCATCATTGTCACTGATGTCATTGCCACTCTGCTCCTTGCTTTGGGAGTCTTCTGCTTTGCTGGACATGAGACTGGAAGGCTGTCTGGGGCTGCCGACACACAAGCTCTGTTGAGGAATGACCAGGTCTATCAGCCCCTCCGAGATCGAGATGATGCTCAGTACAGCCACCTTGGAGGAAACTGGGCTCGGAACAAGTGA (SEQ ID NO: 10)

[0159] Examples of the entire CD3 delta sequences of each nucleic acid and amino acid are shown below (the underlines indicate the signal peptide sequences): ATGGAGCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCAAGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGGGTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAATCCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCACCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCCGGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGCCGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACCAGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGGGCCAGAAACAAG(SEQ ID NO: 35) MEHSTFLSGLVLATLLSQVSP FKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK(SEQ ID NO: 36)

[0160] CD3 gamma (T cell surface glycoprotein CD3 gamma chain gene CD3G P09693) Signal peptide MEQGKGLAVL ILAIILQGTRA(SEQ ID NO: 11)

[0161] Extracellular domain sp|P09693|23-116 QSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATIS(SEQ ID NO: 12)

[0162] Transmembrane domain sp|P09693|117-137 GFLFAEIVSIFVLAVGVYFIA (SEQ ID NO: 13)

[0163] Intracellular domain sp|P09693|138-182 GQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRN (SEQ ID NO: 14)

[0164] Homo sapiens CD3g molecule (CD3G), mRNA; NM_000073.3:81-629 Homo sapiens CD3g molecule (CD3G), mRNA ATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATGATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGAGGGATGTATCAGTGTAAAGGATCACAGAACAAGTCAAAACCACTCCAAGTGTATTACAGAATGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATTGA (SEQ ID NO: 15)

[0165] Examples showing the entire CD3 gamma sequences of each nucleic acid and amino acid are shown below (the underlines indicate signal peptide sequences): ATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATGATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGGGATGTATCAGTGTAAAGGATCACAGAACAAGTCAAAACCACTCCAAGTGTATTACAGAATGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAAT (SEQ ID NO: 33) MEQGKGLAVLILAIILQGTRAQSIKGNHLVKVIDYQEDGSVLTCTCDAEAAKNITWFKDGKMIGFLEDKKWNLGSNAKD PRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDQTRLPNDQLYQPLKDREDDQYSHLQGNQLRN (SEQ ID NO: 34)

[0166] CD3 zeta

[0167] Signal peptide sp|P20963| SP MKWKALFTAAILQAQLPITEA (SEQ ID NO: 16)

[0168] Extracellular domain sp|P20963|22-30 ECD QSFGLLDPK (SEQ ID NO: 17)

[0169] Transmembrane domain sp|P20963|31-51 tmd LCYLDGILFIYGVILTALFL (SEQ ID NO: 18)

[0170] Intracellular domain sp|P20963|52-164 ICD RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (SEQ ID NO: 19)

[0171] Examples showing the entire CD3 zeta sequences of the respective nucleic acids and amino acids are shown below (the underlines indicate signal peptide sequences): ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGCA CAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGC (SEQ ID NO: 31) MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (SEQ ID NO: 32)

[0172] Homo sapiens CD247 molecule (CD247; also known 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 ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCTAA (SEQ ID NO: 20)

[0173] In a specific embodiment, the NK cells are modified to express one or more of the TCRα chain, TCRβ chain, TCRγ chain, and TCRδ chain, and any combination thereof can be utilized. In a specific case, the NK cells are modified to express the T cell receptor (TCR) αβ chain or TCRγδ chain. In a particular case, the NK cells are modified to express only a part or all of the constant region of one or more of the TCRα chain, TCRβ chain, TCRγ chain, and TCRδ chain. The NK cells can be modified to express only a part or all of the constant region of the T cell receptor (TCR) αβ chain or TCRγδ chain. When a part of the constant region is utilized, the part of the constant region can be at least 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, or 400 amino acids, including consecutive amino acids of any constant region. A part of the constant region can include 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 the constant region, including consecutive amino acids of the constant region.

[0174] In a specific case, any sequence included herein is utilized to modify NK cells, while in other cases, sequences related thereto in terms of identity are utilized. For example, related sequences that are at least 80, 85, 90, 95, 96, 97, 98, 99% identical to any sequence included herein can be utilized in the present disclosure.

[0175] Certain constructs for the expression of various TCR / CD3 proteins in NK cells can be utilized in various configurations. In specific instances, NK cells can be transduced or transfected with one or more vectors for expressing any of the various proteins encompassed herein that include at least one or more components of the TCR / CD3 complex. In specific instances, one or more vectors themselves can be multicistronic or not multicistronic by being capable of ultimately generating more than one distinct polypeptide. When one or more multicistronic vectors are employed, they can utilize one or more internal ribosome entry sites (IRES) and / or one or more 2A self-cleaving peptide sites. When one or more 2A sequences are utilized, the following can be used, where GSG is any linker.

[0176] T2A (GSG) EGRGSLLTCGDVEENPGP (SEQ ID NO: 21)

[0177] P2A (GSG) (GSG) ATNFSLLKQAGDVEENPGP (SEQ ID NO: 22)

[0178] E2A (GSG) QCTNYALLKLAGDVESNPGP (SEQ ID NO: 23)

[0179] F2A (GSG) VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 24)

[0180] In situations where multiple protein components are expressed from a multi-cistronic vector, the order on the polynucleotide vector in the 5' to 3' direction can be in any order, but alternatively, they are present on the vector in a specific order. A multi-cistronic vector can express multiple components of the CD3 receptor complex and may or may not express other heterologous proteins, or a multi-cistronic vector can express multiple components of the CD3 receptor complex and one or more other heterologous proteins. A multi-cistronic vector can express multiple components of the TCR receptor complex and may or may not express other heterologous proteins, or a multi-cistronic vector can express multiple components of the TCR receptor complex and one or more other heterologous proteins. A multi-cistronic vector may or may not be able to 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, the multi-cistronic vector comprises one or more components of the CD3 receptor complex and one or more heterologous proteins such as cytokines and engineered antigen receptors, such as CARs.

[0181] Figure 2A shows an example of a multi-cistronic vector in which the full lengths of CD3ε, CD3δ, CD3γ, and CD3ζ are present and separated by the same or different 2A self-cleaving peptide sites. As further noted in the plasmid map of Figure 2B, the multi-cistronic vector can include the signal peptide, extracellular domain, transmembrane domain, and intracellular domain of each of CD3ε, CD3δ, CD3γ, and CD3ζ.

[0182] Figure 3 provides a table showing examples of various TCR expression constructs for engineering NK cells expressing TCRs. In certain embodiments of the present disclosure, the CD3 receptor components and the TCR receptor components are expressed from different vectors in NK cells. In any case, the vector(s) can express a TCR directed against a specific antigen such as a cancer antigen or a viral antigen. The TCR may or may not include at least a portion of CD3ζ, including the intracellular domain of CD3ζ, and further, the NK cells may also express CD3ζ as a molecule separate from the TCR and also as part of the CD3 receptor complex. Similarly, the CAR may or may not include at least a portion of CD3ζ, including the intracellular domain of CD3ζ, and further, the NK cells may also express CD3ζ as a molecule separate from the TCR and also as part of the CD3 receptor complex.

[0183] In a specific embodiment, the TCR of the engineered NK cells is utilized not necessarily as a therapeutic modality of the cells but as a structural support or scaffold for promoting the function or enhanced function of the CD3 receptor complex. That is, the TCR can be any TCR, but is not particularly utilized for its ability to target a desired antigen. In such a case, for example, a TCR targeting a viral antigen can be employed for NK cells used in cancers not necessarily related to that particular virus. In other cases, the TCR is selected for its ability to target a specific cancer antigen. Examples of antigens to which the TCR can be directed are provided elsewhere in this specification.

[0184] Figure 3 depicts the following exemplary configurations:

[0185] TCR1: Refers to a TCRpp65 (TCR against HLA-A2 restricted CMVpp65) linked to an intracellular CD3zeta domain and full-length CD3γ, full-length CD3δ, and full-length CD3ε, and the construct can also be referred to as TCRpp65ZicdGDEFL which may include the following sequences:

[0186] In TCRpp65ZicdGDEFL, the corresponding component sequences are as follows, although these specific sequences or other sequences may be used in this construct and / or other constructs:

[0187] TCRb - extracellular domain: MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGIYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD (SEQ ID NO: 40) ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGACCCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAATGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCCAACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGCCCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGATACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCCGAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGTGTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGGAGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC(SEQ ID NO: 41)

[0188] CD3 zeta intracellular domain (Z-ICD): RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAGDVEENPGP(SEQ ID NO: 42) (where the P2A sequence is at the C-terminus) AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCgccaccaacttctccctgctgaagcaggccggcgacgtggaggagaaccccggcccc(SEQ ID NO: 43)(Here, the lowercase sequence is the P2A sequence)

[0189] TCRa extracellular domain: MILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESS(SEQ ID NO: 44) ATGATCCTGAACGTGGAGCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAACTTCACCTGCAGCTTCCCCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCGCCAAGAGCCCCGAGGCCCTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGGATCAGCGCCACCCTGAACACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCCCGAGGACAGCGCCACCTACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCGGAACCAGCCTGACCGTGATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGAGCCAGAGCAAGGACAGCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGC(SEQ ID NO: 45)

[0190] CD3 gamma-delta-epsilon (CD3GDE): MEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGP(SEQ ID NO: 46)(where the E2A sequence is at the C-terminus)

[0191] IL-15: MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*(SEQ ID NO:48) ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC(SEQ ID NO:49)

[0192] TCR2: Refers to TCR pp65 linked to full-length CD3 zeta, full-length CD3 gamma, full-length CD3 delta, and full-length CD3 epsilon, lacking IL-15. A representative sequence is as follows:

[0193] TCR3: Refers to TCRpp65 that binds to the intracellular CD3z domain and IL-15, and may also be called TCRpp65Zicd15. A representative sequence is as follows: MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGIYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAGDVEENPGPMILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRPGPQCTNYALLKLAGDVESNPGPMRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*(SEQ ID NO: 52)

[0194] In TCRpp65Zicd15, the corresponding component sequences are as follows, although these specific sequences or other sequences may be utilized in this construct and / or other constructs:

[0195] TCRb - extracellular domain: MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGIYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD (SEQ ID NO: 40) ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGACCCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAATGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCCAACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGCCCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGATACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCCGAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGTGTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGGAGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAACGACAGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC(SEQ ID NO: 41)

[0196] CD3 zeta intracellular domain (Z-ICD): RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAGDVEENPGP(SEQ ID NO: 42) (where the P2A sequence is at the C-terminus) AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCgccaccaacttctccctgctgaagcaggccggcgacgtggaggagaaccccggcccc(SEQ ID NO: 43)(Here, the lowercase letters are the P2A sequence)

[0197] TCRa extracellular domain: MILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESS(SEQ ID NO: 44) ATGATCCTGAACGTGGAGCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAACTTCACCTGCAGCTTCCCCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCGCCAAGAGCCCCGAGGCCCTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGGATCAGCGCCACCCTGAACACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCCCGAGGACAGCGCCACCTACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCGGAACCAGCCTGACCGTGATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGAGCCAGAGCAAGGACAGCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGC(SEQ ID NO: 45)

[0198] CD3 zeta intracellular domain (Z-ICD) (in certain embodiments, two or more Z-ICD sequences may be utilized): RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRPGPQCTNYALLKLAGDVESNPGP(SEQ ID NO: 53) AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC (SEQ ID NO: 54)

[0199] IL-15: MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*(SEQ ID NO: 48) ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC(SEQ ID NO:49)

[0200] TCR4: Refers to TCRpp65, also called TCRpp65βα, and a representative sequence is as follows: MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGIYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAGDVEENPGPMILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRPGPQCTNYALLKLAGDVESNPGPMRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*(SEQ ID NO: 55)

[0201] For TCRpp65βα, the corresponding component sequences are as follows, although these specific sequences or other sequences may be used in this construct and / or other constructs:

[0202] TCRb - extracellular domain: MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGIYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD (SEQ ID NO: 40) ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGACCCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAATGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCCAACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGCCCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGATACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCCGAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGTGTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGGAGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC(SEQ ID NO: 41)

[0203] CD3 zeta intracellular domain (Z-ICD): RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRATNFSLLKQAGDVEENPGP(SEQ ID NO: 42) AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC (SEQ ID NO: 54)

[0204] TCRa extracellular domain: MILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESS(SEQ ID NO: 44) ATGATCCTGAACGTGGAGCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAACTTCACCTGCAGCTTCCCCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCGCCAAGAGCCCCGAGGCCCTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGGATCAGCGCCACCCTGAACACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCCCGAGGACAGCGCCACCTACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCGGAACCAGCCTGACCGTGATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGAGCCAGAGCAAGGACAGCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGC(SEQ ID NO: 45)

[0205] CD3 zeta intracellular domain (Z-ICD): RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRPGPQCTNYALLKLAGDVESNPGP(SEQ ID NO: 53) AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC (SEQ ID NO: 54)

[0206] IL-15: MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*(SEQ ID NO: 48) ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC(SEQ ID NO:49)

[0207] A representative sequence of TCRpp65βα is as follows: MDSWTFCCVSLCILVAKHTDAGVIQSPRHEVTEMGQEVTLRCKPISGHDYLFWYRQTMMRGLELLIYFNNNVPIDDSGMPEDRFSAKMPNASFSTLKIQPSEPRDSAVYFCASSSANYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDFEGRGSLLTCGDVEENPGPMLLEHLLIILWMQLTWVSGQQLNQSPQSMFIQEGEDVSMNCTSSSIFNTWLWYKQDPGEGPVLLIALYKAGELTSNGRLTAQFGITRKDSFLNISASIPSDVGIYFCAGPMKTSYDKVIFGPGTSLSVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS*(SEQ ID NO:57)

[0208] Z1: Refers to full-length CD3zeta, full-length CD3γ, full-length CD3δ, and full-length CD3ε linked to IL15 (see FIGS. 2A and 2B), and may also be referred to as CD3ZFLGDEFL15. A representative sequence is as follows: MLEMKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGPMRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO: 58)

[0209] Z2 refers to the one in which full-length CD3zeta, full-length CD3γ, full-length CD3δ, full-length CD3ε are combined with membrane-bound IL21 (membrane-bound IL21 has a CD8 transmembrane domain), and is also called CD3ZGDEFLSP821CD28. The representative sequence is as follows:

[0210] In the case of CD3ZGDEFLSP821CD28, the corresponding component sequences are as follows, although these specific sequences or other sequences may be used for this construct and / or other constructs:

[0211] CD3: MLEMKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGP (SEQ ID NO: 61)

[0212] SP CD8: MRICLTSDRLAPAAGLAAPRRQAV (SEQ ID NO: 63) Atgcgcatttgcctgaccagcgatcgcctggcgccggcggcgggcctggcggcgccgcgccgccaggcggtg (SEQ ID NO: 64)

[0213] IL-21: HKSSSQGQDRHMIRMRQLIDIVDQLKNYVNDLVPEFLPAPEDVETNCEWSAFSCFQKAQLKSANTGNNERIINVSIKKLKRKPPSTNAGRRQKHRLTCPSCDSYEKKPPKEFLERFKSLLQKMIHQHLSSRTHGSEDS (SEQ ID NO: 65) CATAAATCTTCCTCTCAAGGTCAGGACCGCCATATGATTCGAATGCGGCAGCTGATTGACATAGTCGATCAACTGAAGAACTATGTGAATGATCTTGTGCCCGAGTTTTTGCCAGCCCCTGAAGACGTAGAAACTAATTGTGAGTGGAGTGCCTTTTCCTGCTTTCAAAAGGCACAGCTGAAATCCGCCAACACGGGCAATAACGAACGGATAATTAACGTATCCATTAAGAAGCTGAAGCGGAAGCCGCCCTCAACCAATGCGGGACGGCGGCAAAAGCATCGCTTGACCTGTCCGTCATGCGACAGCTACGAGAAAAAGCCCCCGAAGGAGTTCTTGGAACGCTTCAAGAGTCTCCTTCAGAAAATGATTCACCAGCACCTGTCCTCACGGACGCACGGAAGCGAGGACAGT (SEQ ID NO: 66)

[0214] CD8 hinge: TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 67) ACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACCATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCGCCTGTGAT (SEQ ID NO: 68)

[0215] CD28 transmembrane domain: FWVLVVVGGVLACYSLLVTVAFIIFWV* (SEQ ID NO: 69) TTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTG (SEQ ID NO: 70)

[0216] Z3: Refers to full-length CD3zeta, full-length CD3γ, full-length CD3δ, and full-length CD3ε that are bound to membrane-bound IL21 (membrane-bound IL21 has a CD28 transmembrane domain), and a representative sequence is as follows, also called CD3ZGDEFL8SP21CD8:

[0217] For CD3ZGDEFL8SP21CD8, the corresponding component sequences are as follows, although these specific sequences or other sequences may be utilized in this construct and / or other constructs:

[0218] CD3: MLEMKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGP (SEQ ID NO: 61)

[0219] SP CD8: MRICLTSDRLAPAAGLAAPRRQAV (SEQ ID NO: 63) Atgcgcatttgcctgaccagcgatcgcctggcgccggcggcgggcctggcggcgccgcgccgccaggcggtg (SEQ ID NO: 64)

[0220] IL-21: HKSSSQGQDRHMIRMRQLIDIVDQLKNYVNDLVPEFLPAPEDVETNCEWSAFSCFQKAQLKSANTGNNERIINVSIKKLKRKPPSTNAGRRQKHRLTCPSCDSYEKKPPKEFLERFKSLLQKMIHQHLSSRTHGSEDS (SEQ ID NO: 65) Cataaatcttcctctcaaggtcaggaccgccatatgattcgaatgcggcagctgattgacatagtcgatcaactgaagaactatgtgaatgatcttgtgcccgagtttttgccagcccctgaagacgtagaaactaattgtgagtggagtgccttttcctgctttcaaaaggcacagctgaaatccgccaacacgggcaataacgaacggataattaacgtatccattaagaagctgaagcggaagccgccctcaaccaatgcgggacggcggcaaaagcatcgcttgacctgtccgtcatgcgacagctacgagaaaaagcccccgaaggagttcttggaacgcttcaagagtctccttcagaaaatgattcaccagcacctgtcctcacggacgcacggaagcgaggacagt (SEQ ID NO: 66)

[0221] CD8 hinge: TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 67) ACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACCATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCGCCTGTGAT (SEQ ID NO: 68)

[0222] CD8 transmembrane domain: IYIWAPLAGTCGVLLLSLVIT* (SEQ ID NO: 72) ATCTACATCTGGGCGCCCTTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTGGTTATCACC (SEQ ID NO: 73)

[0223] In certain embodiments, provided herein are CD3 constructs comprising a fusogen having an intracellular costimulatory domain derived from CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, DNAM, or any combination thereof. In certain embodiments, the intracellular costimulatory domain is fused to CD3δ, CD3ε, CD3γ, and / or CD3ζ. In certain embodiments, such CD3 fusion constructs comprise CD3ζ fused to the DAP10 intracellular costimulatory domain. In certain embodiments, such CD3 fusion constructs comprise CD3ζ fused to the CD28 intracellular costimulatory domain. In certain embodiments, such CD3 fusion constructs comprise CD3ζ fused to both the DAP10 intracellular costimulatory domain and the CD28 intracellular costimulatory domain. In certain embodiments, CD3ζ fused to the DAP10 intracellular costimulatory 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, CD3ζ fused to the CD28 intracellular costimulatory 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, CD3ζ fused to both the DAP10 intracellular costimulatory domain and the CD28 intracellular costimulatory 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, CD3ζ fused to the 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, CD3ζ fused to the 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, CD3ζ fused to the DAP10 intracellular co-stimulatory domain and the 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: 111. In certain embodiments, CD3ζ fused to the intracellular domain may not include the C-terminal 2A domain. In certain embodiments, CD3ζ fused to the intracellular domain may not include the N-terminal signal peptide domain. ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGCTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC(SEQ ID NO: 106) ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCAAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC(SEQ ID NO: 107) ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCACTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC(SEQ ID NO: 108) MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLLCARPRRSPAQEDGKVYINMPGRGRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGP(SEQ ID NO: 109) MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGP (SEQ ID NO: 110) MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMPGRGRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGP (SEQ ID NO: 111)

[0224] In certain embodiments, the DAP10 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, the 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, the DAP10 intracellular co-stimulatory domain and the CD28 intracellular co-stimulatory domain are 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, the 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, the 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, the DAP10 intracellular co-stimulatory domain and the CD28 intracellular co-stimulatory domain are 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. CTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGC (SEQ ID NO: 112) AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCA (SEQ ID NO: 113) AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCACTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGC (SEQ ID NO: 114) LCARPRRSPAQEDGKVYINMPGRG (SEQ ID NO: 115) RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS (SEQ ID NO: 116) RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMPGRG (SEQ ID NO: 117)

[0225] UTNK15-DAP10 refers to full-length CD3 zeta, full-length CD3 gamma, full-length CD3 delta, and full-length CD3 epsilon linked to IL-15, which contains a fusion with an intracellular co-stimulatory domain derived from DAP10, and can 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, the UTNK15-DAP10 amino acid sequence can 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.

[0226] UTNK15-28 refers to full-length CD zeta, full-length CD3 gamma, full-length CD3 delta, and full-length CD3 epsilon linked to IL-15, which contains a fusion with an intracellular co-stimulatory domain derived from CD28, and can 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, the UTNK15-28 amino acid sequence can 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.

[0227] UTNK15-28-DAP10 refers to full-length CD3 zeta, full-length CD3 gamma, full-length CD3 delta, and full-length CD3 epsilon linked to IL15, which contains a fusion having an intracellular co-stimulatory domain derived from DAP10 and an intracellular co-stimulatory domain derived from CD28, and can 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, the UTNK15-28-DAP10 amino acid sequence can 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.

[0228] As described in FIG. 3 and as mentioned above, the term "linked" refers to being present on the same polynucleotide vector and does not necessarily mean that two polypeptides are expressed as one polypeptide. For example, cytokines produced from the vectors of the present disclosure can ultimately be produced as molecules separate from any one or more TCR / CD3 receptor complex components. On the other hand, the term "fused" or "fusion" refers to two polypeptides that include a peptide bond that joins the two molecules, i.e., two polypeptides that are covalently bonded by an amide bond and are not separated by a cleavage element such as a 2A element.

[0229] One specific example of a TCR that can be utilized intracellularly is the NY-ESO TCR, and specific examples of the sequence include at least the following:

[0230] TCRα: XQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVRPLYGGSYIPTFGRGTSLIVHPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 25)

[0231] TCRβ: GVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVLEDLKNVFPPKVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG (SEQ ID NO: 26)

[0232] In certain embodiments, the TCR can comprise a TCR α-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. aaacaggaggtgacacagattcctgcagctctgagtgtcccagaaggagaaaacttggttctcaactgcagtttcact gatagcgctatttacaac ctccagtggtttaggcaggaccctgggaaaggtctcacatctctgttgctt attcagtcaagtcagagagag caaacaagtggaagacttaatgcctcgctggataaatcatcaggacgtagtactttatacattgcagcttctcagcctggtgactcagccacctacctc tgtgctgtgaggcccctttatggaggaagctacatacctacattt ggaagaggaaccagccttattgttcatccgtat (SEQ ID NO: 85)

[0233] In certain embodiments, the TCR can include a constant region of the TCRα 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: 86. atccagaaccctgaccctgccgtgtaccagctgagagactctaaatccagtgacaagtctgtctgcctattcaccgattttgattctcaaacaaatgtgtcacaaagtaaggattctgatgtgtatatcacagacaaaactgtgctagacatgaggtctatggacttcaagagcaacagtgctgtggcctggagcaacaaatctgactttgcatgtgcaaacgccttcaacaacagcattattccagaagacaccttcttccccagcccagaaagttcctgtgatgtcaagctggtcgagaaaagctttgaaacagatacgaacctaaactttcaaaacctgtcagtgattgggttccgaatcctcctcctgaaagtggccgggtttaatctgctcatgacgctgcggctgtggtccagc (SEQ ID NO: 86)

[0234] In certain embodiments, the TCR can include a TCRα 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. atggagaccctcttgggcctgcttatcctttggctgcagctgcaatgggtgagcagcaaacaggaggtgacacagattcctgcagctctgagtgtcccagaaggagaaaacttggttctcaactgcagtttcact gatagcgctatttacaacctccagtggtttaggcaggaccctgggaaaggtctcacatctctgttgctt attcagtcaagtcagagagag caaacaagtggaagacttaatgcctcgctggataaatcatcaggacgtagtactttatacattgcagcttctcagcctggtgactcagccacctacctc tgtgctgtgaggcccctttatggaggaagctacatacctacattt ggaagaggaaccagccttattgttcatccgtatatccagaaccctgaccctgccgtgtaccagctgagagactctaaatccagtgacaagtctgtctgcctattcaccgattttgattctcaaacaaatgtgtcacaaagtaaggattctgatgtgtatatcacagacaaaactgtgctagacatgaggtctatggacttcaagagcaacagtgctgtggcctggagcaacaaatctgactttgcatgtgcaaacgccttcaacaacagcattattccagaagacaccttcttccccagcccagaaagttcctgtgatgtcaagctggtcgagaaaagctttgaaacagatacgaacctaaactttcaaaacctgtcagtgattgggttccgaatcctcctcctgaaagtggccgggtttaatctgctcatgacgctgcggctgtggtccagc(SEQ ID NO: 87)

[0235] In certain embodiments, the TCR may comprise a TCR α-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. XQEVTQIPAALSVPEGENLVLNCSFT DSAIYN LQWFRQDPGKGLTSLLL IQSSQRE QTSGRLNASLDKSSGRSTLYIAASQPGDSATYL CAVRPLYGGSYIPTF GRGTSLIVHPY(SEQ ID NO: 88)

[0236] In certain embodiments, the TCR may comprise a TCR α-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. IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 89)

[0237] In certain embodiments, the TCR may comprise an α-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)

[0238] In certain embodiments, the TCR may comprise an α-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)

[0239] In certain embodiments, the TCR may comprise an α-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)

[0240] In certain embodiments, the TCR may comprise a TCR β-chain variant 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. ggtgtcactcagaccccaaaattccaggtcctgaagacaggacagagcatgacactgcagtgtgcccaggat atgaaccatgaatac atgtcctggtatcgacaagacccaggcatggggctgaggctgattcattac tcagttggtgctggtatc actgaccaaggagaagtccccaatggctacaatgtctccagatcaaccacagaggatttcccgctcaggctgctgtcggctgctccctcccagacatctgtgtacttc tgtgccagcagttacgtcgggaacaccggggagctgtttttt ggagaaggctctaggctgaccgtactggag (SEQ ID NO: 93)

[0241] In certain embodiments, the TCR may include a constant region of the TCRβ 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: 94. Gacctgaaaaacgtgttcccacccaaggtcgctgtgtttgagccatcagaagcagagatctcccacacccaaaaggccacactggtatgcctggccacaggcttctaccccgaccacgtggagctgagctggtgggtgaatgggaaggaggtgcacagtggggtcagcacagacccgcagcccctcaaggagcagcccgccctcaatgactccagatactgcctgagcagccgcctgagggtctcggccaccttctggcagaacccccgcaaccacttccgctgtcaagtccagttctacgggctctcggagaatgacgagtggacccaggatagggccaaacccgtcacccagatcgtcagcgccgaggcctggggtagagcagactgtggcttcacctccgagtcttaccagcaaggggtcctgtctgccaccatcctctatgagatcttgctagggaaggccaccttgtatgccgtgctggtcagtgccctcgtgctgatggccatggtcaagagaaaggattccagaggc(SEQ ID NO: 94)

[0242] In certain embodiments, the TCR may comprise a TCRβ 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. Atgagcatcggcctcctgtgctgtgcagccttgtctctcctgtgggcaggtccagtgaatgctggtgtcactcagaccccaaaattccaggtcctgaagacaggacagagcatgacactgcagtgtgcccaggat atgaaccatgaatac atgtcctggtatcgacaagacccaggcatggggctgaggctgattcattac tcagttggtgctggtatcactgaccaaggagaagtccccaatggctacaatgtctccagatcaaccacagaggatttcccgctcaggctgctgtcggctgctccctcccagacatctgtgtacttc tgtgccagcagttacgtcgggaacaccggggagctgtttttt ggagaaggctctaggctgaccgtactggaggacctgaaaaacgtgttcccacccAaggtcgctgtgtttgagccatcagaagcagagatctcccacacccaaaaggccacactggtatgcctggccacaggcttctaccccgaccacgtggagctgagctggtgggtgaatgggaaggaggtgcacagtggggtcagcacagacccgcagcccctcaaggagcagcccgccctcaatgactccagatactgcctgagcagccgcctgagggtctcggccaccttctggcagaacccccgcaaccacttccgctgtcaagtccagttctacgggctctcggagaatgacgagtggacccaggatagggccaaacccgtcacccagatcgtcagcgccgaggcctggggtagagcagactgtggcttcacctccgagtcttaccagcaaggggtcctgtctgccaccatcctctatgagatcttgctagggaaggccaccttgtatgccgtgctggtcagtgccctcgtgctgatggccatggtcaagagaaaggattccagaggc(SEQ ID NO: 95)

[0243] In certain embodiments, the TCR may comprise a TCRβ 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. GVTQTPKFQVLKTGQSMTLQCAQD MNHEY MSWYRQDPGMGLRLIHY SVGAGI TDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFF GEGSRLTVLE (SEQ ID NO: 96)

[0244] In certain embodiments, the TCR may comprise a TCRβ 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. DLKNVFPPKVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG (SEQ ID NO: 97)

[0245] In certain embodiments, the TCR may comprise a β-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)

[0246] In certain embodiments, the TCR may comprise a β-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)

[0247] In certain embodiments, the TCR may comprise a β-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)

[0248] In certain embodiments, the TCR (e.g., TCRα, β, δ, and / or γ) chain may include a signal peptide. In certain embodiments, the 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, the 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) atgagcatcggcctcctgtgctgtgcagccttgtctctcctgtgggcaggtccagtgaatgct(SEQ ID NO: 102) METLLGLLILWLQLQWVSS(SEQ ID NO: 103) MSIGLLCCAALSLLWAGPVNA(SEQ ID NO: 104)

[0249] In certain embodiments, the TCR recognizes a peptide corresponding to amino acid residues 157-165 of human cancer testis Ag NY-ESO-1 in the context of the HLA-A*02 class I allele. In certain embodiments, the TCR may target an epitope characterized by the amino acid sequence according to SEQ ID NO: 105. SLLMWITQC(SEQ ID NO: 105)

[0250] One specific example of a TCR that can be utilized intracellularly is TCRpp65α, and specific examples of the sequence include at least the following (the underlines indicate the signal peptide sequence): ATGGACTCCTGGACCTTCTGCTGTGTGTCCCTTTGCATCCTGGTAGCAAAGCACACAGATGCTGGACAACAGCTGAATCAGAGTCCTCAATCTATGTTTATCCAGGAAGGAGAAGATGTCTCCATGAACTGCACTTCTTCAAGCATATTTAACACCTGGCTATGGTACAAGCAGGACCCTGGGGAAGGTCCTGTCCTCTTGATAGCCTTATATAAGGCTGGTGAATTGACCTCAAATGGAAGACTGACTGCTCAGTTTGGTATAACCAGAAAGGACAGCTTCCTGAATATCTCAGCATCCATACCCAGTGATGTAGGCATCTACTTCTGTGCTGGACCCATGAAAACCTCCTACGACAAGGTGATATTTGGGCCAGGGACAAGCTTATCAGTCATTCCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGC(SEQ ID NO: 27) MDSWTFCCVSLCILVAKHTDAGQQLNQSPQSMFIQEGEDVSMNCTSSSIFNTWLWYKQDPGEGPVLLIALYKAGELTSNGRLTAQFGITRKDSFLNISASIPSDVGIYFCAGPMKTSYDKVIFGPGTSLSVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 28)

[0251] One specific example of a TCR that can be utilized intracellularly is TCRpp65β, and specific examples of the sequence include at least the following (the underlines indicate signal peptide sequences): ATGGACTCCTGGACCTTCTGCTGTGTGTCCCTTTGCATCCTGGTAGCAAAGCACACAGATGCTGGAGTTATCCAGTCACCCCGGCACGAGGTGACAGAGATGGGACAAGAAGTGACTCTGAGATGTAAACCAATTTCAGGACACGACTACCTTTTCTGGTACAGACAGACCATGATGCGGGGACTGGAGTTGCTCATTTACTTTAACAACAACGTTCCGATAGATGATTCAGGGATGCCCGAGGATCGATTCTCAGCTAAGATGCCTAATGCATCATTCTCCACTCTGAAGATCCAGCCCTCAGAACCCAGGGACTCAGCTGTGTACTTCTGTGCCAGCAGTTCGGCAAACTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGTAGAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCCGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTTACCTCGGTGTCCTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAGGCCACCCTGTATGCTGTGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAAGAGAAAGGATTTC (SEQ ID NO: 29) MDSWTFCCVSLCILVAKHTDAGVIQSPRHEVTEMGQEVTLRCKPISGHDYLFWYRQTMMRGLELLIYFNNNVPIDDSGMPEDRFSAKMPNASFSTLKIQPSEPRDSAVYFCASSSANYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 30)

[0252] TCRpp65ZFLGDEFL15

[0253] In certain embodiments, constructs in which TCRpp65 is linked to full-length CD3ζ, full-length CD3γ, full-length CD3δ, full-length CD3ε, and further linked to IL-15 can be utilized (which can be referred to as TCRpp65ZFLGDEFL15). A representative sequence of such a construct is as follows:

[0254] In TCRpp65ZFLGDEFL15, the corresponding component sequences are as follows, although these specific sequences or other sequences may be utilized in this construct and / or other constructs:

[0255] TCRb - extracellular domain: MLEGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSPVTGGIYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADATNFSLLKQAGDVEENPGP (SEQ ID NO: 75) (and including a P2A sequence at its C-terminus) ATGCTCGAGGGAGTGACCCAGACCCCCAAGTTCCAGGTGCTGAAGACCGGACAGAGCATGACCCTGCAGTGCGCCCAGGACATGAACCACGAGTACATGAGCTGGTACCGGCAGGACCCCGGAATGGGACTGCGGCTGATCCACTACAGCGTGGGAGCCGGAATCACCGACCAGGGAGAGGTGCCCAACGGATACAACGTGAGCCGGAGCACCACCGAGGACTTCCCCCTGCGGCTGCTGAGCGCCGCCCCCAGCCAGACCAGCGTGTACTTCTGCGCCAGCAGCCCCGTGACCGGAGGAATCTACGGATACACCTTCGGAAGCGGAACCCGGCTGACCGTGGTGGAGGACCTGAACAAGGTGTTCCCCCCCGAGGTGGCCGTGTTCGAGCCCAGCGAGGCCGAGATCAGCCACACCCAGAAGGCCACCCTGGTGTGCCTGGCCACCGGATTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAACGGAAAGGAGGTGCACAGCGGAGTGAGCACCGACCCCCAGCCCCTGAAGGAGCAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCCGGCTGCGGGTGAGCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCCGGTGCCAGGTGCAGTTCTACGGACTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGAGCGCCGAGGCCTGGGGACGGGCCGAC(SEQ ID NO: 76)

[0256] TCRa extracellular domain: MILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCARNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDAYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSEGRGSLLTCGDVEENPGP (SEQ ID NO: 77) (and including the T2A sequence at its C-terminus) ATGATCCTGAACGTGGAGCAGAGCCCCCAGAGCCTGCACGTGCAGGAGGGAGACAGCACCAACTTCACCTGCAGCTTCCCCAGCAGCAACTTCTACGCCCTGCACTGGTACCGGTGGGAGACCGCCAAGAGCCCCGAGGCCCTGTTCGTGATGACCCTGAACGGAGACGAGAAGAAGAAGGGACGGATCAGCGCCACCCTGAACACCAAGGAGGGATACAGCTACCTGTACATCAAGGGAAGCCAGCCCGAGGACAGCGCCACCTACCTGTGCGCCCGGAACACCGGAAACCAGTTCTACTTCGGAACCGGAACCAGCCTGACCGTGATCCCCAACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGAGCCAGAGCAAGGACAGCGACGCCTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGAGCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACCTTCTTCCCCAGCCCCGAGAGCAGCGCCACCAACTTCTCCCTGCTGAAGCAGGCCGGCGACGTGGAGGAGAACCCCGGCCCC (SEQ ID NO: 78)

[0257] TCR5, called TCR Cγδ ZFLGDEFL15, is in the constant regions of TCR gamma and delta, and is linked to full-length CD3 zeta, full-length CD3 gamma, full-length CD3 delta, full-length CD3 epsilon; and IL-15. Representative sequences are as follows:

[0258] TCR constant gamma delta (TCR Cγδ) MRWALLVLLAFLSPASQDKQLDADVSPKPTIFLPSIAETKLQKAGTYLCLLEKFFPDIIKIHWQEKKSNTILGSQEGNTMKTNDTYMKFSWLTVPEESLDKEHRCIVRHENNKNGIDQEIIFPPIKTDVTTVDPKYNYSKDANDVITMDPKDNWSKDANDTLLLQLTNTSAYYTYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMILTVGFSFLFFYRGTLCSQPHTKPSVFVMKNGTNVACLVKEFYPKDIRINLVSSKKITEFDPAIVISPSGKYNAVKLGKYEDSNSVTCSVQHDNKTVHSTDFEVKTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFL (SEQ ID NO: 82)

[0259] CD3: MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGP(SEQ ID NO:79)

[0260] IL-15: MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*(SEQ ID NO:48) ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC(SEQ ID NO:49)

[0261] TCR6: Also called TCRCabZFLGDEFL15, it is linked to the constant regions of TCRα and β, full-length CD3zeta, full-length CD3γ, full-length CD3δ, full-length CD3ε; and IL-15. A representative sequence is as follows:

[0262] TCR constant αβ (TCRCab) METLLGLLILWLQLQWVSSIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSGATNFSLLKQAGDVEENPGPMSIGLLCCAALSLLWAGPVNADLKNVFPPKVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG(SEQ ID NO: 83)

[0263] CD3: MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGP (SEQ ID NO: 79)

[0264] IL-15: MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*(SEQ ID NO: 48) ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC(SEQ ID NO: 49)

[0265] In some embodiments, the TCR construct comprises an NY-ESO-specific TCR and a CD8 alpha / beta coreceptor 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 2A element (e.g., a 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., an 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 coreceptor 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 coreceptor 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.

[0266] In some embodiments, the CD8 alpha coreceptor molecule is transcriptionally linked to any TCR molecule disclosed herein. In some embodiments, the CD8 alpha coreceptor 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, the CD8 beta coreceptor 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, the CD8 alpha coreceptor 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, the CD8 beta coreceptor 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. ATGAGGCCACGACTTTGGCTGCTGCTCGCTGCACAGTTGACTGTACTGCATGGCAATAGTGTGTTGCAGCAGACACCTGCATACATCAAGGTTCAGACAAATAAGATGGTTATGCTGAGTTGCGAGGCAAAAATTAGTTTGAGCAATATGCGGATCTACTGGTTGCGACAGAGACAGGCTCCCAGTAGTGATAGTCACCACGAATTCCTGGCTCTTTGGGATTCCGCAAAAGGAACGATTCATGGGGAAGAAGTAGAGCAGGAGAAGATTGCGGTTTTCCGCGATGCATCTCGCTTTATCCTTAATCTTACATCCGTTAAGCCTGAGGACAGTGGGATCTATTTTTGTATGATTGTAGGGTCCCCCGAATTGACATTTGGGAAGGGTACGCAGCTCTCCGTAGTTGACTTTCTGCCCACAACGGCACAACCCACTAAGAAGTCCACCCTGAAGAAGCGCGTCTGTCGCTTGCCCAGACCTGAAACCCAAAAGGGTCCACTCTGTTCCCCTATAACCCTGGGGTTGTTGGTGGCGGGCGTCTTGGTCCTGCTTGTTAGCTTGGGCGTAGCCATTCATCTGTGTTGCCGAAGACGCAGAGCCCGACTTAGATTTATGAAGCAATTCTATAAGTGA(SEQ ID NO: 126) ATGGCCTTGCCCGTCACTGCGCTTTTGCTCCCGCTCGCTCTTCTCCTGCATGCAGCCCGACCATCTCAATTTAGAGTTTCTCCACTCGACAGGACGTGGAACCTCGGCGAAACCGTCGAACTTAAATGTCAAGTACTTCTCTCAAATCCGACTTCTGGTTGCTCATGGCTCTTTCAGCCGAGAGGAGCAGCTGCCAGCCCCACCTTCCTGCTGTATCTCTCCCAGAACAAGCCGAAGGCCGCCGAAGGGCTCGATACTCAACGATTTAGCGGGAAGCGACTCGGGGACACGTTCGTTCTTACTCTCAGCGATTTTAGAAGAGAGAACGAGGGATATTATTTTTGTTCCGCACTCTCTAACAGCATCATGTACTTCAGTCATTTTGTACCAGTCTTTCTCCCTGCAAAACCAACGACTACTCCAGCACCAAGACCGCCCACTCCCGCACCTACTATTGCAAGCCAACCTTTGAGTCTCCGACCAGAGGCATGCAGACCTGCTGCTGGAGGTGCAGTACATACGCGAGGGTTGGATTTTGCCTGCGATATCTATATCTGGGCCCCCTTGGCCGGCACGTGCGGGGTGCTCCTGCTGAGTCTCGTAATTACTCTTTATTGTAATCATAGAAACCGCAGAAGGGTGTGTAAGTGTCCCCGGCCTGTCGTGAAAAGCGGGGATAAGCCCAGTTTGTCTGCTCGGTACGTC(SEQ ID NO: 127) MRPRLWLLLAAQLTVLHGNSVLQQTPAYIKVQTNKMVMLSCEAKISLSNMRIYWLRQRQAPSSDSHHEFLALWDSAKGTIHGEEVEQEKIAVFRDASRFILNLTSVKPEDSGIYFCMIVGSPELTFGKGTQLSVVDFLPTTAQPTKKSTLKKRVCRLPRPETQKGPLCSPITLGLLVAGVLVLLVSLGVAIHLCCRRRRARLRFMKQFYK(SEQ ID NO: 128) MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPRGAAASPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGYYFCSALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV(SEQ ID NO: 129)

[0267] In some embodiments, the TCR construct comprises a PRAME - specific TCR chain. In some embodiments, a TCR construct comprising a PRAME - specific TCR chain comprises TCR alpha and beta chains found in PRAME - specific TCR clone 46, clone 54, and / or clone DSK3. In some embodiments, a TCR construct comprising a PRAME - specific TCR chain comprises TCR alpha and beta chains that target the PRAME epitopes SLLQHLIGL (SEQ ID NO: 131) and / or QLLALLPSL (SEQ ID NO: 132).

[0268] In some embodiments, a TCR construct comprising a PRAME - specific TCR chain 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 a PRAME - specific TCR chain 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). ATGCTTCTGGAACACCTGCTGATTATCCTGTGGATGCAACTCACGTGGGTCTCCGGGCAACAACTGAATCAAAGCCCCCAATCCATGTTTATACAGGAGGGAGAGGACGTAAGTATGAATTGCACATCTTCATCTATCTTTAACACCTGGCTGTGGTACAAACAAGACCCCGGAGAAGGTCCTGTACTTCTCATCGCACTTTACAAAGCAGGTGAGCTTACCAGTAACGGGAGACTCACCGCACAGTTCGGTATTACAAGAAAGGATTCCTTTCTCAACATCTCCGCTTCTATCCCTTCAGACGTCGGAATTTATTTTTGTGCTGGTATCCCTCGAGACAATTACGGTCAAAACTTTGTATTTGGGCCTGGGACTCGGCTGTCAGTTTTGCCGTATATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAGAAGTCCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGTCCGTGATCGGCTTCAGAATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGC(SEQ ID NO: 133) ATGGGCATTAGGCTGCTGTGCAGAGTAGCATTTTGCTTTCTGGCAGTAGGATTGGTCGATGTAAAGGTTACACAGTCCTCACGGTACTTGGTAAAGCGCACTGGTGAAAAGGTCTTTCTGGAATGTGTACAAGATATGGATCACGAAAATATGTTTTGGTACAGGCAAGATCCCGGCCTTGGACTTAGACTGATATATTTCTCCTACGATGTTAAAATGAAGGAGAAGGGCGATATTCCAGAAGGATATTCCGTGAGCCGCGAAAAGAAGGAGCGATTCAGTTTGATACTCGAAAGTGCCTCCACAAACCAAACCTCTATGTACCTTTGCGCGTCAACGCCGTGGCTGGCCGGTGGCAATGAACAATTCTTCGGGCCGGGTACGCGCCTCACTGTCCTGGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTTGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAATGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC(SEQ ID NO: 134) MLLEHLLIILWMQLTWVSGQQLNQSPQSMFIQEGEDVSMNCTSSSIFNTWLWYKQDPGEGPVLLIALYKAGELTSNGRLTAQFGITRKDSFLNISASIPSDVGIYFCAGIPRDNYGQNFVFGPGTRLSVLPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 135) MGIRLLCRVAFCFLAVGLVDVKVTQSSRYLVKRTGEKVFLECVQDMDHENMFWYRQDPGLGLRLIYFSYDVKMKEKGDIPEGYSVSREKKERFSLILESASTNQTSMYLCASTPWLAGGNEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG(SEQ ID NO: 136)

[0269] In some embodiments, a TCR construct comprising a PRAME-specific TCR chain 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 a PRAME-specific TCR chain 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). ATGCTGCTGCTGCTGGTGCCCGTGCTGGAAGTGATCTTCACCCTGGGCGGCACCAGAGCCCAGAGCGTGACACAGCTGGGCAGCCACGTGTCCGTGTCTGAGAGGGCCCTGGTGCTGCTGAGATGCAACTACTCTTCTAGCGTGCCCCCCTACCTGTTTTGGTACGTGCAGTACCCCAACCAGGGGCTGCAGCTGCTCCTGAAGTACACCAGCGCCGCCACACTGGTGAAGGGCATCAACGGCTTCGAGGCCGAGTTCAAGAAGTCCGAGACAAGCTTCCACCTGACCAAGCCCAGCGCCCACATGTCTGACGCCGCCGAGTACTTCTGTGCCGTGAGCGGCCAGACCGGCGCCAACAACCTGTTCTTCGGCACCGGCACCCGGCTGACAGTGATCCCTTACATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAGAAGTCCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGTCCGTGATCGGCTTCAGAATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGC(SEQ ID NO: 137) ATGGGCTTCCGGCTGCTGTGCTGCGTGGCCTTTTGTCTGCTGGGAGCCGGACCTGTGGATAGCGGCGTGACCCAGACCCCCAAGCACCTGATCACCGCCACCGGCCAGAGAGTGACCCTGCGCTGCAGCCCTAGAAGCGGCGACCTGAGCGTGTACTGGTATCAGCAGAGCCTCGACCAGGGCCTGCAGTTCCTGATCCAGTACTACAACGGCGAGGAACGGGCCAAGGGCAACATCCTGGAACGGTTCAGCGCCCAGCAGTTCCCCGATCTGCACAGCGAGCTGAACCTGAGCAGCCTGGAACTGGGCGACAGCGCCCTGTACTTCTGCGCCAGCGCCAGATGGGATAGAGGCGGCGAGCAGTACTTCGGCCCTGGCACCAGACTGACCGTGACCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTTGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAATGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC(SEQ ID NO: 138) MLLLLVPVLEVIFTLGGTRAQSVTQLGSHVSVSERALVLLRCNYSSSVPPYLFWYVQYPNQGLQLLLKYTSAATLVKGINGFEAEFKKSETSFHLTKPSAHMSDAAEYFCAVSGQTGANNLFFGTGTRLTVIPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 139) MGFRLLCCVAFCLLGAGPVDSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASARWDRGGEQYFGPGTRLTVTEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG(SEQ ID NO: 140)

[0270] In some embodiments, a TCR construct comprising a PRAME-specific TCR chain 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 a PRAME-specific TCR chain 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). ATGAAGAGCCTGAGGGTACTGCTGGTGATATTGTGGCTTCAGCTTAGTTGGGTCTGGTCACAACAAAAGGAAGTTGAGCAAAACTCAGGACCACTGAGTGTACCCGAGGGCGCTATAGCATCACTGAACTGTACCTACTCAGATCGGGGAAGCCAATCCTTTTTCTGGTACAGACAGTATTCCGGGAAGAGTCCTGAGTTGATCATGTTTATATACTCCAATGGCGATAAGGAGGATGGACGCTTCACCGCTCAGCTTAATAAAGCGTCACAGTATGTATCCCTCCTGATTCGGGACTCACAACCATCTGACTCTGCAACATACCTTTGTGCCGTAAAGGACAACGCCGGGAACATGCTCACTTTTGGAGGAGGTACCCGGCTTATGGTAAAACCACATATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAGAAGTCCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGTCCGTGATCGGCTTCAGAATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGC(SEQ ID NO: 141) MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVKDNAGNMLTFGGGTRLMVKPHIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 142) ATGGGATTCCGGCTTCTTTGTTGTGTGGCATTTTGTCTGTTGGGTGCGGGTCCAGTCGATAGTGGTGTAACTCAGACACCAAAACACCTTATCACGGCAACTGGGCAACGAGTGACGCTCCGCTGTAGCCCGAGGTCCGGTGATTTGAGTGTGTACTGGTACCAGCAATCTTTGGACCAGGGCTTGCAGTTCCTCATACAGTATTACAATGGTGAAGAAAGAGCGAAGGGTAATATCCTGGAAAGATTCTCCGCACAACAGTTTCCTGATCTCCACAGCGAACTGAACCTGAGTTCTCTCGAGCTCGGGGATAGTGCTTTGTACTTCTGCGCGTCATCCGACGGTGGCGGAGTCTATGAACAATATTTCGGCCCAGGGACTAGGCTTACGGTGACGGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTTGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAATGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC(SEQ ID NO: 143) MGFRLLCCVAFCLLGAGPVDSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASSDGGGVYEQYFGPGTRLTVTEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG(SEQ ID NO: 144)

[0271] In some embodiments, a TCR construct comprising a PRAME-specific TCR chain 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 a PRAME-specific TCR chain comprises a TCR alpha chain and a TCR beta chain found in the PRAME-specific TCR clone T116-49 and / or T402-93 and / or modified versions thereof. In some embodiments, a TCR construct comprising a PRAME-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the PRAME epitope LYVDSLFFL (SEQ ID NO: 167). In some embodiments, the PRAME-specific TCR sequence, TCR variable domain sequence, CDR sequence, and / or TCR constant domain sequence are described in International Publication No. WO2022 / 063966A1, which is incorporated herein by reference for the purposes described herein. In some embodiments, a TCR construct comprising a PRAME-specific TCR chain 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. ATGGAGACACTGCTGAAGGTGCTGTCTGGCACACTGCTGTGGCAGCTGACCTGGGTCCGATCTCAGCAGCCTGTTCAGTCTCCTCAGGCCGTGATCCTGAGAGAAGGCGAGGACGCCGTGATCAACTGCAGCAGCTCTAAGGCCCTGTACAGCGTGCACTGGTACAGACAGAAGCACGGCGAGGCCCCTGTGTTCCTGATGATCCTGCTGAAAGGCGGCGAGCAGAAGGGCCACGAGAAGATCAGCGCCAGCTTCAACGAGAAGAAGCAGCAGTCCAGCCTGTACCTGACAGCCAGCCAGCTGAGCTACAGCGGCACCTACTTTTGCGGCACAGCCAATAGCGGCGGCAGCAACTACAAGCTGACCTTCGGCAAGGGCACCCTGCTGACCGTGAATCCCAAT(SEQ ID NO: 145) ATGCTGCTGATCACCTCCATGCTGGTGCTGTGGATGCAGCTGAGCCAAGTGAACGGCCAGCAAGTGATGCAGATCCCTCAGTACCAGCACGTGCAAGAAGGCGAGGACTTCACCACCTACTGCAACAGCAGCACCACACTGAGCAACATCCAGTGGTACAAGCAGCGGCCTGGCGGACACCCTGTGTTTCTGATCCAGCTGGTCAAGTCCGGCGAAGTGAAGAAGCAGAAGCGGCTGACCTTCCAGTTCGGCGAGGCCAAGAAGAACAGCAGCCTGCACATCACCGCCACACAGACCACCGATGTGGGCACCTACTTTTGTGCTGGCGCCCTGCCTAGAGCCGGCAGCTATCAACTGACATTCGGCAAGGGCACCAAGCTGAGCGTGATCCCCAAC(SEQ ID NO: 146) ATGGAGACACTGCTGAAGGTGCTGTCTGGCACACTGCTGTGGCAGCTGACCTGGGTCCGATCTCAGCAGCCTGTTCAGTCTCCTCAGGCCGTGATCCTGAGAGAAGGCGAGGACGCCGTGATCAACTGCAGCAGCTCTAAGGCCCTGTACAGCGTGCACTGGTACAGACAGAAGCACGGCGAGGCCCCTGTGTTCCTGATGATCCTGCTGAAAGGCGGCGAGCAGAAGGGCCACGAGAAGATCAGCGCCAGCTTCAACGAGAAGAAGCAGCAGTCCAGCCTGTACCTGACAGCCAGCCAGCTGAGCTACAGCGGCACCTACTTTTGCGGCACAGCCAATAGCGGCGGCAGCAACTACAAGCTGACCTTCGGCAAGGGCACCCTGCTGACCGTGAATCCCAATATCCAGAATCCGGAGCCCGCCGTATACCAGCTGAAGGACCCTAGAAGCCAGGACAGCACCCTGTGCCTGTTCACCGACTTCGACAGCCAGATCAACGTGCCCAAGACCATGGAAAGCGGCACCTTCATCACCGACAAGACAGTGCTGGACATGAAGGCCATGGACAGCAAGTCCAACGGCGCAATCGCCTGGTCCAACCAGACCAGCTTCACATGCCAGGACATCTTCAAAGAGACAAACGCCACATACCCCAGCAGCGACGTGCCCTGTGATGCCACCCTGACAGAGAAGTCCTTCGAGACAGACATGAACCTGAACTTCCAGAATCTGTCCGTGATGGGCCTGAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAATCTGCTGATGACCCTGCGGCTGTGGTCCAGC(SEQ ID NO: 147) ATGCTGCTGATCACCTCCATGCTGGTGCTGTGGATGCAGCTGAGCCAAGTGAACGGCCAGCAAGTGATGCAGATCCCTCAGTACCAGCACGTGCAAGAAGGCGAGGACTTCACCACCTACTGCAACAGCAGCACCACACTGAGCAACATCCAGTGGTACAAGCAGCGGCCTGGCGGACACCCTGTGTTTCTGATCCAGCTGGTCAAGTCCGGCGAAGTGAAGAAGCAGAAGCGGCTGACCTTCCAGTTCGGCGAGGCCAAGAAGAACAGCAGCCTGCACATCACCGCCACACAGACCACCGATGTGGGCACCTACTTTTGTGCTGGCGCCCTGCCTAGAGCCGGCAGCTATCAACTGACATTCGGCAAGGGCACCAAGCTGAGCGTGATCCCCAACATCCAGAATCCGGAGCCCGCCGTATACCAGCTGAAGGACCCTAGAAGCCAGGACAGCACCCTGTGCCTGTTCACCGACTTCGACAGCCAGATCAACGTGCCCAAGACCATGGAAAGCGGCACCTTCATCACCGACAAGACAGTGCTGGACATGAAGGCCATGGACAGCAAGTCCAACGGCGCAATCGCCTGGTCCAACCAGACCAGCTTCACATGCCAGGACATCTTCAAAGAGACAAACGCCACATACCCCAGCAGCGACGTGCCCTGTGATGCCACCCTGACAGAGAAGTCCTTCGAGACAGACATGAACCTGAACTTCCAGAATCTGTCCGTGATGGGCCTGAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAATCTGCTGATGACCCTGCGGCTGTGGTCCAGC(SEQ ID NO: 148) ATGGGCACCAGACTGTTCTTCTACGTGGCCCTGTGTCTGCTGTGGACAGGCCATGTGGATGCCGGAATCACACAGAGCCCCAGACACAAAGTGACCGAGACAGGCACCCCTGTGACACTGAGATGTCACCAGACCGAGAACCATCGGTACATGTATTGGTACAGACAGGACCCCGGCCACGGCCTGAGACTGATCCACTATAGCTACGGCGTGAAGGACACCGACAAGGGCGAAGTGTCTGACGGCTACAGCGTGTCCAGAAGCAAGACCGAGGACTTCCTGCTGACCCTGGAAAGCGCCACAAGCAGCCAGACCAGCGTGTACTTCTGCGCCATCAGCGACTACGAGGGCACCGAGGCCTTTTTTGGCCAAGGCACAAGACTGACCGTGGTG(SEQ ID NO: 149) ATGCTGTGTTCTCTGCTGGCTCTGCTGCTGGGCACCTTTTTTGGCGTCAGAAGCCAGACCATCCACCAGTGGCCTGCTACACTGGTGCAGCCTGTTGGAAGCCCTCTGAGCCTGGAATGTACCGTGGAAGGCACCAGCAATCCCAACCTGTACTGGTACAGACAGGCCGCTGGAAGAGGACTGCAGCTGCTGTTTTACAGCGTCGGCATCGGCCAGATCAGCAGCGAGGTTCCACAGAATCTGAGCGCCAGCAGACCCCAGGACAGACAGTTTATCCTGAGCAGCAAGAAGCTGCTGCTGAGCGACAGCGGCTTCTACCTGTGTGCTTGGAGCCTCGGAGCCGGCTACACCGACACACAGTATTTTGGCCCTGGCACCAGACTGACCGTGCTG(SEQ ID NO: 150) ATGGGCACCAGACTGTTCTTCTACGTGGCCCTGTGTCTGCTGTGGACAGGCCATGTGGATGCCGGAATCACACAGAGCCCCAGACACAAAGTGACCGAGACAGGCACCCCTGTGACACTGAGATGTCACCAGACCGAGAACCATCGGTACATGTATTGGTACAGACAGGACCCCGGCCACGGCCTGAGACTGATCCACTATAGCTACGGCGTGAAGGACACCGACAAGGGCGAAGTGTCTGACGGCTACAGCGTGTCCAGAAGCAAGACCGAGGACTTCCTGCTGACCCTGGAAAGCGCCACAAGCAGCCAGACCAGCGTGTACTTCTGCGCCATCAGCGACTACGAGGGCACCGAGGCCTTTTTTGGCCAAGGCACAAGACTGACCGTGGTGGAAGATCTCCGGAACGTGACCCCCCCTAAAGTGACCCTGTTCGAACCCAGCAAGGCCGAGATCGCCAACAAGCAGAAAGCCACCCTCGTGTGCCTGGCCAGAGGCTTCTTCCCCGACCATGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGAGTGTCCACCGACCCTCAGGCCTACAAAGAGAGCAACTACAGCTACTGCCTGAGCAGCAGACTGCGGGTGTCCGCCACCTTCTGGCACAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTTCACGGCCTGAGCGAAGAGGACAAGTGGCCCGAAGGCTCCCCCAAGCCCGTGACCCAGAATATCTCTGCCGAGGCCTGGGGCAGAGCCGACTGTGGAATTACCAGCGCCAGCTACCACCAGGGCGTGCTGTCTGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCTGGCCTGGTGCTGATGGCCATGGTCAAGAAGAAGAACAGC(SEQ ID NO: 151) ATGCTGTGTTCTCTGCTGGCTCTGCTGCTGGGCACCTTTTTTGGCGTCAGAAGCCAGACCATCCACCAGTGGCCTGCTACACTGGTGCAGCCTGTTGGAAGCCCTCTGAGCCTGGAATGTACCGTGGAAGGCACCAGCAATCCCAACCTGTACTGGTACAGACAGGCCGCTGGAAGAGGACTGCAGCTGCTGTTTTACAGCGTCGGCATCGGCCAGATCAGCAGCGAGGTTCCACAGAATCTGAGCGCCAGCAGACCCCAGGACAGACAGTTTATCCTGAGCAGCAAGAAGCTGCTGCTGAGCGACAGCGGCTTCTACCTGTGTGCTTGGAGCCTCGGAGCCGGCTACACCGACACACAGTATTTTGGCCCTGGCACCAGACTGACCGTGCTGGAAGATCTCCGGAACGTGACCCCCCCTAAAGTGACCCTGTTCGAACCCAGCAAGGCCGAGATCGCCAACAAGCAGAAAGCCACCCTCGTGTGCCTGGCCAGAGGCTTCTTCCCCGACCATGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGAGTGTCCACCGACCCTCAGGCCTACAAAGAGAGCAACTACAGCTACTGCCTGAGCAGCAGACTGCGGGTGTCCGCCACCTTCTGGCACAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTTCACGGCCTGAGCGAAGAGGACAAGTGGCCCGAAGGCTCCCCCAAGCCCGTGACCCAGAATATCTCTGCCGAGGCCTGGGGCAGAGCCGACTGTGGAATTACCAGCGCCAGCTACCACCAGGGCGTGCTGTCTGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCTGGCCTGGTGCTGATGGCCATGGTCAAGAAGAAGAACAGC(SEQ ID NO: 152) METLLKVLSGTLLWQLTWVRSQQPVQSPQAVILREGEDAVINCSSSKALYSVHWYRQKHGEAPVFLMILLKGGEQKGHEKISASFNEKKQQSSLYLTASQLSYSGTYFCGTANSGGSNYKLTFGKGTLLTVNPN(SEQ ID NO: 153) MLLITSMLVLWMQLSQVNGQQVMQIPQYQHVQEGEDFTTYCNSSTTLSNIQWYKQRPGGHPVFLIQLVKSGEVKKQKRLTFQFGEAKKNSSLHITATQTTDVGTYFCAGALPRAGSYQLTFGKGTKLSVIPN(SEQ ID NO: 154) IQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 155) IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 156) IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSSDVPCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 157) METLLKVLSGTLLWQLTWVRSQQPVQSPQAVILREGEDAVINCSSSKALYSVHWYRQKHGEAPVFLMILLKGGEQKGHEKISASFNEKKQQSSLYLTASQLSYSGTYFCGTANSGGSNYKLTFGKGTLLTVNPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 158) MLLITSMLVLWMQLSQVNGQQVMQIPQYQHVQEGEDFTTYCNSSTTLSNIQWYKQRPGGHPVFLIQLVKSGEVKKQKRLTFQFGEAKKNSSLHITATQTTDVGTYFCAGALPRAGSYQLTFGKGTKLSVIPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 159) MGTRLFFYVALCLLWTGHVDAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGLRLIHYSYGVKDTDKGEVSDGYSVSRSKTEDFLLTLESATSSQTSVYFCAISDYEGTEAFFGQGTRLTVV(SEQ ID NO: 160) MLCSLLALLLGTFFGVRSQTIHQWPATLVQPVGSPLSLECTVEGTSNPNLYWYRQAAGRGLQLLFYSVGIGQISSEVPQNLSASRPQDRQFILSSKKLLLSDSGFYLCAWSLGAGYTDTQYFGPGTRLTVL(SEQ ID NO: 161) EDLRNVTPPKVTLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYHQGVLSATILYEILLGKATLYAVLVSGLVLMAMVKKKNS (SEQ ID NO: 162) DLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 163) EDLNKVFPPEVAVFEPSKAEIAHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGITSASYHQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 164) MGTRLFFYVALCLLWTGHVDAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGLRLIHYSYGVKDTDKGEVSDGYSVSRSKTEDFLLTLESATSSQTSVYFCAISDYEGTEAFFGQGTRLTVVEDLRNVTPPKVTLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYHQGVLSATILYEILLGKATLYAVLVSGLVLMAMVKKKNS (SEQ ID NO: 165) MLCSLLALLLGTFFGVRSQTIHQWPATLVQPVGSPLSLECTVEGTSNPNLYWYRQAAGRGLQLLFYSVGIGQISSEVPQNLSASRPQDRQFILSSKKLLLSDSGFYLCAWSLGAGYTDTQYFGPGTRLTVLEDLRNVTPPKVTLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYHQGVLSATILYEILLGKATLYAVLVSGLVLMAMVKKKNS(SEQ ID NO: 166)

[0272] In some embodiments, the TCR construct comprises a gp100-specific TCR chain. In some embodiments, a TCR construct comprising a gp100-specific TCR chain comprises a TCR alpha chain and a TCR beta chain found in the gp100-specific TCR clone Sp(0.01)A and / or a modified version thereof. In some embodiments, a TCR construct comprising a gp100-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the gp100 epitope KTWGQYWQV (SEQ ID NO: 168). In some embodiments, the gp100-specific TCR sequence, TCR variable domain sequence, CDR sequence, and / or TCR constant domain sequence are described in U.S. Patent Publication No. 8,216,565B2, which is incorporated herein by reference for the purposes described herein.

[0273] In some embodiments, the TCR construct comprising a gp100-specific TCR chain 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, the TCR construct comprising a gp100-specific TCR chain 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. ATGAAATCCTTGAGTGTTTCCCTAGTGGTCCTGTGGCTCCAGTTAAACTGGGTGAACAGCCAGCAGAAGGTGCAGCAGAGCCCAGAATCCCTCATTGTCCCAGAGGGAGCCATGACCTCTCTCAACTGCACTTTCAGCGACAGTGCTTCTCAGTATTTTGCATGGTACAGACAGCATTCTGGGAAAGCCCCCAAGGCACTGATGTCCATCTTCTCCAATGGTGAAAAAGAAGAAGGCAGATTCACAATTCACCTCAATAAAGCCAGTCTGCATTTCTCGCTACACATCAGAGACTCCCAGCCCAGTGACTCTGCTCTCTACCTCTGTGCAGCCAATAACTATGCCCAGGGATTAACCTTCGGTCTTGGCACCAGAGTATCTGTGTTTCCCTACATCCAGAACCCAGAACCTGCTGTGTACCAGTTAAAAGATCCTCGGTCTCAGGACAGCACCCTCTGCCTGTTCACCGACTTTGACTCCCAAATCAATGTGCCGAAAACCATGGAATCTGGAACGTTCATCACTGACAAAACTGTGCTGGACATGAAAGCTATGGATTCCAAGAGCAATGGGGCCATTGCCTGGAGCAACCAGACAAGCTTCACCTGCCAAGATATCTTCAAAGAGACCAACGCCACCTACCCCAGTTCAGACGTTCCCTGTGATGCCACGTTGACTGAGAAAAGCTTTGAAACAGATATGAACCTAAACTTTCAAAACCTGTCAGTTATGGGACTCCGAATCCTCCTGCTGAAAGTAGCCGGATTTAACCTGCTCATGACGCTGAGGCTGTGGTCCAGTTGA(SEQ ID NO: 169) ATGGGCTCCAGACTCTTCTTTGTGGTTTTGATTCTCCTGTGTGCAAAACACATGGAGGCTGCAGTCACCCAAAGTCCAAGAAGCAAGGTGGCAGTAACAGGAGGAAAGGTGACATTGAGCTGTCACCAGACTAATAACCATGACTATATGTACTGGTATCGGCAGGACACGGGGCATGGGCTGAGGCTGATCCATTACTCATATGTCGCTGACAGCACGGAGAAAGGAGATATCCCTGATGGGTACAAGGCCTCCAGACCAAGCCAAGAGAATTTCTCTCTCATTCTGGAGTTGGCTTCCCTTTCTCAGACAGCTGTATATTTCTGTGCCAGCAGCCCTGGGGGGGGGGGGGAACAGTACTTCGGTCCCGGCACCAGGCTCACGGTTTTAGAGGATCTGAGAAATGTGACTCCACCCAAGGTCTCCTTGTTTGAGCCATCAAAAGCAGAGATTGCAAACAAACGAAAGGCTACCCTCGTGTGCTTGGCCAGGGGCTTCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGCAAGGAGGTCCACAGTGGGGTCAGCACGGACCCTCAGGCCTACAAGGAGAGCAATTATAGCTACTGCCTGAGCAGCCGCCTGAGGGTCTCTGCTACCTTCTGGCACAATCCTCGAAACCACTTCCGCTGCCAAGTGCAGTTCCATGGGCTTTCAGAGGAGGACAAGTGGCCAGAGGGCTCACCCAAACCTGTCACACAGAACATCAGTGCAGAGGCCTGGGGCCGAGCAGACTGTGGGATTACCTCAGCATCCTATCAACAAGGGGTCTTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAAGCCACCCTGTATGCTGTGCTTGTCAGTACACTGGTGGTGATGGCTATGGTCAAAAGAAAGAATTCATGA(SEQ ID NO: 170) MKSLSVSLVVLWLQLNWVNSQQKVQQSPESLIVPEGAMTSLNCTFSDSASQYFAWYRQHSGKAPKALMSIFSNGEKEEGRFTIHLNKASLHFSLHIRDSQPSDSALYLCAANNYAQGLTFGLGTRVSVFPYIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 171) MGSRLFFVVLILLCAKHMEAAVTQSPRSKVAVTGGKVTLSCHQTNNHDYMYWYRQDTGHGLRLIHYSYVADSTEKGDIPDGYKASRPSQENFSLILELASLSQTAVYFCASSPGGGGEQYFGPGTRLTVLEDLRNVTPPKVSLFEPSKAEIANKRKATLVCLARGFFPDHVELSWWVNGKEVHSGVSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVMAMVKRKNS(SEQ ID NO: 172) QQKVQQSPESLIVPEGAMTSLNCTFSDSASQYFAWYRQHSGKAPKALMSIFSNGEKEEGRFTIHLNKASLHFSLHIRDSQPSDSALYLCAANNYAQGLTFGLGTRVSVFPY(SEQ ID NO: 173) EAAVTQSPRSKVAVTGGKVTLSCHQTNNHDYMYWYRQDTGHGLRLIHYSYVADSTEKGDIPDGYKASRPSQENFSLILELASLSQTAVYFCASSPGGGGEQYFGPGTRLTVL(SEQ ID NO: 174)

[0274] In some embodiments, the TCR construct comprises a MART-1 specific TCR chain. In some embodiments, a TCR construct comprising a MART-1 specific TCR chain comprises a TCR alpha chain and a TCR beta chain found in MART-1 specific TCR clone F4 and / or F5 and / or modified versions thereof. In some embodiments, a TCR construct comprising a MART-1 specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the MART-1 epitope AAGIGILTV (SEQ ID NO: 175). In some embodiments, the MART-1 specific TCR sequence, TCR variable domain sequence, CDR sequence, and / or TCR constant domain sequence are described in U.S. Patent Publication No. 9,128,080 B2, which is incorporated herein by reference for the purposes described herein.

[0275] In some embodiments, a TCR construct comprising a MART-1 specific TCR chain 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 a MART-1 specific TCR chain 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. ATGTTGCTTGAACATTTATTAATAATCTTGTGGATGCAGCTGACATGGGTCAGTGGTCAACAGCTGAATCAGAGTCCTCAATCTATGTTTATCCAGGAAGGAGAAGATGTCTCCATGAACTGCACTTCTTCAAGCATATTTAACACCTGGCTATGGTACAAGCAGGACCCTGGGGAAGGTCCTGTCCTCTTGATAGCCTTATATAAGGCTGGTGAATTGACCTCAAATGGAAGACTGACTGCTCAGTTTGGTATAACCAGAAAGGACAGCTTCCTGAATATCTCAGCATCCATACCTAGTGATGTAGGCATCTACTTCTGTGCTGGTGGGACCGGTAACCAGTTCTATTTTGGGACAGGGACAAGTTTGACGGTCATTCCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAGGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGC(SEQ ID NO: 176) ATGGGCACAAGGTTGTTCTTCTATGTGGCCCTTTGTCTCCTGTGGACAGGACACATGGATGCTGGAATCACCCAGAGCCCAAGACACAAGGTCACAGAGACAGGAACACCAGTGACTCTGAGATGTCACCAGACTGAGAACCACCGCTATATGTACTGGTATCGACAAGACCCGGGGCATGGGCTGAGGCTGATCCATTACTCATATGGTGTTAAAGATACTGACAAAGGAGAAGTCTCAGATGGCTATAGTGTCTCTAGATCAAAGACAGAGGATTTCCTCCTCACTCTGGAGTCCGCTACCAGCTCCCAGACATCTGTGTACTTCTGTGCCATCAGTGAGGTAGGGGTTGGGCAGCCCCAGCATTTTGGTGATGGGACTCGACTCTCCATCCTAGAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCCGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCATGTGGCTTTACCTCGTCCTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAGGCCACCCTGTATGCTGTGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAAGAGAAAGGATTTC(SEQ ID NO: 177) ATGATGAAATCCTTGAGAGTTTTACTAGTGATCCTGTGGCTTCAGTTGAGCTGGGTTTGGAGCCAACAGAAGGAGGTGGAGCAGAATTCTGGACCCCTCAGTGTTCCAGAGGGAGCCATTGCCTCTCTCAACTGCACTTACAGTGACCGAGGTTCCCAGTCCTTCTTCTGGTACAGACAATATTCTGGGAAAAGCCCTGAGTTGATAATGTTCATATACTCCAATGGTGACAAAGAAGATGGAAGGTTTACAGCACAGCTCAATAAAGCCAGCCAGTATGTTTCTCTGCTCATCAGAGACTCCCAGCCCAGTGATTCAGCCACCTACCTCTGTGCCGTGAACTTCGGAGGAGGAAAGCTTATCTTCGGACAGGGAACGGAGTTATCTGTGAAACCCAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 178) ATGAGAATCAGGCTCCTGTGCTGTGTGGCCTTTTCTCTCCTGTGGGCAGGTCCAGTGATTGCTGGGATCACCCAGGCACCAACATCTCAGATCCTGGCAGCAGGACGGCGCATGACACTGAGATGTACCCAGGATATGAGACATAATGCCATGTACTGGTATAGACAAGATCTAGGACTGGGGCTAAGGCTCATCCATTATTCAAATACTGCAGGTACCACTGGCAAAGGAGAAGTCCCTGATGGTTATAGTGTCTCCAGAGCAAACACAGATGATTTCCCCCTCACGTTGGCGTCTGCTGTACCCTCTCAGACATCTGTGTACTTCTGTGCCAGCAGCCTAAGTTTCGGCACTGAAGCTTTCTTTGGACAAGGCACCAGACTCACAGTTGTAGAGGACCTGAACAAGGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTTCCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACGGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCCGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCATGTGGCTTTACCTCGTCCTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAGGCCACCCTGTATGCTGTGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAAGAGAAAGGATTTC(SEQ ID NO: 179) GQQLNQSPQSMFIQEGEDVSMNCTSSSIFNTWLWYKQDPGEGPVLLIALYKAGELTSNGRLTAQFGITRKDSFLNISASIPSDVGIYFCAGGTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 180) DAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGLRLIHYSYGVKDTDKGEVSDGYSVSRSKTEDFLLTLESATSSQTSVYFCAISEVGVGQPQHFGDGTRLSILEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRACGFTSSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF(SEQ ID NO: 181) QKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVNFGGGKLIFGQGTELSVKPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 182) IAGITQAPTSQILAAGRRMTLRCTQDMRHNAMYWYRQDLGLGLRLIHYSNTAGTTGKGEVPDGYSVSRANTDDFPLTLASAVPSQTSVYFCASSLSFGTEAFFGQGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRACGFTSSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF(SEQ ID NO: 183)

[0276] In some embodiments, the TCR construct comprises a tyrosine kinase-specific TCR chain. In some embodiments, a TCR construct comprising a tyrosine kinase-specific TCR chain comprises a TCR alpha chain and a TCR beta chain found in the tyrosine kinase-specific TCR clone TIL 1383I and / or a modified version thereof. In some embodiments, a TCR construct comprising a tyrosine kinase-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target a tyrosine kinase epitope represented by amino acids 368-376 of tyrosine kinase (reactive against the class I MHC (HLA-A2)-restricted epitope (368-376) of tyrosine kinase). In some embodiments, the tyrosine kinase-specific TCR sequence, TCR variable domain sequence, CDR sequence, and / or TCR constant domain sequence are described in the publication Roszkowski et al., Cancer Res. 65(4):1570-6 (2005), which is incorporated herein by reference for the purposes described herein.

[0277] In some embodiments, the TCR construct comprises a MAGE-A3-specific TCR chain. In some embodiments, a TCR construct comprising a MAGE-A3-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target amino acids 271-279 of MAGE-A3, for example, the epitope FLWGPRALV (SEQ ID NO: 184). In some embodiments, a TCR construct comprising a MAGE-A3-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target amino acids 112-120 of MAGE-A3, for example, the epitope KVAELVHFL (SEQ ID NO: 185). In some embodiments, the MAGE-A3-specific TCR sequence, TCR variable domain sequence, CDR sequence, and / or TCR constant domain sequence are described in International Publication No. WO2012 / 054825A1, which is incorporated herein by reference for the purposes described herein. In certain embodiments, the anti-MAGE-A3 112-120 TCR comprises an A118T substitution relative to the wild type (position 118 of the alpha chain is threonine). In certain embodiments, the anti-MAGE-A3 112-120 TCR comprises an A118V substitution relative to the wild type (position 118 of the alpha chain is valine).

[0278] In some embodiments, a TCR construct comprising a MAGE-A3-specific TCR chain 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 a MAGE-A3-specific TCR chain 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. ATGGGTCCTGTCACCTGCTCAGTTCTTGTGCTCCTCCTAATGCTCAGGAGGAGCAATGGCGATGGAGACTCCGTGACCCAGACAGAAGGCCTGGTCACTCTCACAGAAGGGTTGCCTGTGATGCTGAACTGCACCTATCAGACTATTTACTCAAATCCTTTCCTTTTCTGGTATGTGCAACATCTCAATGAATCCCCTCGGCTACTCCTGAAGAGCTTCACAGACAACAAGAGGACCGAGCACCAAGGGTTCCACGCCACTCTCCATAAGAGCAGCAGCTCCTTCCATCTGCAGAAGTCCTCAGCGCAGCTGTCAGACTCTGCCCTGTACTACTGTGCTTTCGACACAAATGCTTACAAAGTCATCTTT(SEQ ID NO: 186) ATGAGAGTTAGGCTCATCTCTGCTGTGGTGCTGTGTTCCCTAGGAACAGGCCTTGTGGACATGAAAGTAACCCAGATGCCAAGATACCTGATCAAAAGAATGGGAGAGAATGTTTTGCTGGAATGTGGACAGGACATGAGCCATGAAACAATGTACTGGTATCGACAAGACCCTGGTCTGGGGCTACAGCTGATTTATATCTCATACGATGTTGATAGTAACAGCGAAGGAGACATCCCTAAAGGATACAGGGTCTCACGGAAGAAGCGGGAGCATTTCTCCCTGATTCTGGATTCTGCTAAAACAAACCAGACATCTGTGTACTTCTGTGCTAGCAGTTCAACAAACACAGAAGTCTTCTTT(SEQ ID NO: 187) ATGGGTCCTGTCACCTGCTCAGTTCTTGTGCTCCTCCTAATGCTCAGGAGGAGCAATGGCGATGGAGACTCCGTGACCCAGACAGAAGGCCTGGTCACTCTCACAGAAGGGTTGCCTGTGATGCTGAACTGCACCTATCAGACTATTTACTCAAATCCTTTCCTTTTCTGGTATGTGCAACATCTCAATGAATCCCCTCGGCTACTCCTGAAGAGCTTCACAGACAACAAGAGGACCGAGCACCAAGGGTTCCACGCCACTCTCCATAAGAGCAGCAGCTCCTTCCATCTGCAGAAGTCCTCAGCGCAGCTGTCAGACTCTGCCCTGTACTACTGTGCTTTCGACACAAATGCTTACAAAGTCATCTTTGGAAAAGGGACACATCTTCATGTTCTCCCTAACATCCAGAACCCAGAACCTGCTGTGTACCAGTTAAAAGATCCTCGGTCTCAGGACAGCACCCTCTGCCTGTTCACCGACTTTGACTCCCAAATCAATGTGCCGAAAACCATGGAATCTGGAACGTTCATCACTGACAAAACTGTGCTGGACATGAAAGCTATGGATTCCAAGAGCAATGGGGCCATTGCCTGGAGCAACCAGACAAGCTTCACCTGCCAAGATATCTTCAAAGAGACCAACACCACCTACCCCAGTTCAGACGTTCCCTGTGATGCCACGTTGACTGAGAAAAGCTTTGAAACAGATATGAACCTAAACTTTCAAAACCTGTCAGTTATGGGACTCCGAATCCTCCTGCTGAAAGTAGCCGGATTTAACCTGCTCATGACGCTGAGGCTGTGGTCCAGTTGA(SEQ ID NO: 188) ATGAGAGTTAGGCTCATCTCTGCTGTGGTGCTGTGTTCCCTAGGAACAGGCCTTGTGGACATGAAAGTAACCCAGATGCCAAGATACCTGATCAAAAGAATGGGAGAGAATGTTTTGCTGGAATGTGGACAGGACATGAGCCATGAAACAATGTACTGGTATCGACAAGACCCTGGTCTGGGGCTACAGCTGATTTATATCTCATACGATGTTGATAGTAACAGCGAAGGAGACATCCCTAAAGGATACAGGGTCTCACGGAAGAAGCGGGAGCATTTCTCCCTGATTCTGGATTCTGCTAAAACAAACCAGACATCTGTGTACTTCTGTGCTAGCAGTTCAACAAACACAGAAGTCTTCTTTGGTAAAGGAACCAGACTCACAGTTGTAGAGGATCTGAGAAATGTGACTCCACCCAAGGTCTCCTTGTTTGAGCCATCAAAAGCAGAGATTGCAAACAAACAAAAGGCTACCCTCGTGTGCTTGGCCAGGGGCTTCTTCCCTGACCACGTGGAGCTGAGCTGGTGGGTGAATGGCAAGGAGGTCCACAGTGGGGTCAGCACGGACCCTCAGGCCTACAAGGAGAGCAATTATAGCTACTGCCTGAGCAGCCGCCTGAGGGTCTCTGCTACCTTCTGGCACAATCCTCGCAACCACTTCCGCTGCCAAGTGCAGTTCCATGGGCTTTCAGAGGAGGACAAGTGGCCAGAGGGCTCACCCAAACCTGTCACACAGAACATCAGTGCAGAGGCCTGGGGCCGAGCAGACTGTGGGATTACCTCAGCATCCTATCAACAAGGGGTCTTGTCTGCCACCATCCTCTATGAGATCCTGCTAGGGAAAGCCACCCTGTATGCTGTGCTTGTCAGTACACTGGTGGTGATGGCTATGGTCAAAAGAAAGAACTCGTGA (SEQ ID NO: 189) ATGGTCCTAGTGACCATTCTGCTGCTCAGCGCGTTCTTCTCACTGAGAGGAAACAGTGCCCAGTCCGTGGACCAGCCTGATGCTCATGTCACGCTCTCTGAAGGAGCCTCCCTGGAGCTCAGATGCAGTTATTCATACAGTGCAGCACCTTACCTCTTCTGGTACGTGCAGTATCCTGGCCAGAGCCTCCAGTTTCTCCTCAAATACATCACAGGAGACACCGTTGTTAAAGGCACCAAGGGCTTTGAGGCCGAGTTTAGGAAGAGTAACTCCTCTTTCAACCTGAAGAAATCCCCAGCCCATTGGAGCGACTCAGCCAAGTACTTCTGTGCACTGGAGGGCCCGGATACAGGAAACTACAAATACGTCTT(SEQ ID NO: 190) ATGGGCATCCAGACCCTCTGTTGTGTGATCTTTTATGTTCTGATAGCAAATCACACAGATGCTGGAGTTACCCAGACACCCAGACATGAGGTGGCAGAGAAAGGACAAACAATAATCCTGAAGTGTGAGCCAGTTTCAGGCCACAATGACCTTTTCTGGTACAGACAGACCAAGATACAGGGACTAGAGTTGCTGAGCTACTTCCGCAGCAAGTCTCTTATGGAAGATGGTGGGGCTTTCAAGGATCGATTCAAAGCTGAGATGCTAAATTCATCCTTCTCCACTCTGAAGATTCAACCTACAGAACCCAGGGACTCAGCTGTGTATCTGTGTGCCAGCAGTTTTGGGACAGCTAGTGCAGAAACGCTGTATTTT(SEQ ID NO: 191) ATGGTCCTAGTGACCATTCTGCTGCTCAGCGCGTTCTTCTCACTGAGAGGAAACAGTGCCCAGTCCGTGGACCAGCCTGATGCTCATGTCACGCTCTCTGAAGGAGCCTCCCTGGAGCTCAGATGCAGTTATTCATACAGTGCAGCACCTTACCTCTTCTGGTACGTGCAGTATCCTGGCCAGAGCCTCCAGTTTCTCCTCAAATACATCACAGGAGACACCGTTGTTAAAGGCACCAAGGGCTTTGAGGCCGAGTTTAGGAAGAGTAACTCCTCTTTCAACCTGAAGAAATCCCCAGCCCATTGGAGCGACTCAGCCAAGTACTTCTGTGCACTGGAGGGCCCGGATACAGGAAACTACAAATACGTCTTTGGAGCAGGTACCAGACTGAAGGTTATAGCACACATCCAGAACCCAGAACCTGCTGTGTACCAGTTAAAAGATCCTCGGTCTCAGGACAGCACCCTCTGCCTGTTCACCGACTTTGACTCCCAAATCAATGTGCCGAAAACCATGGAATCTGGAACGTTCATCACTGACAAAACTGTGCTGGACATGAAAGCTATGGATTCCAAGAGCAATGGGGCCATTGCCTGGAGCAACCAGACAAGCTTCACCTGCCAAGATATCTTCAAAGAGACCAACGCCACCTACCCCAGTTCAGACGTTCCCTGTGATGCCACGTTGACTGAGAAAAGCTTTGAAACAGATATGAACCTAAACTTCCAAAACCTGTCAGTTATGGGACTCCGAATCCTCCTGCTGAAAGTAGCCGGATTTAACCTGCTCATGACGCTGAGGCTGTGGTCCAGTTGA(SEQ ID NO: 192) ATGGGCATCCAGACCCTCTGTTGTGTGATCTTTTATGTTCTGATAGCAAATCACACAGATGCTGGAGTTACCCAGACACCCAGACATGAGGTGGCAGAGAAAGGACAAACAATAATCCTGAAGTGTGAGCCAGTTTCAGGCCACAATGACCTTTTCTGGTACAGACAGACCAAGATACAGGGACTAGAGTTGCTGAGCTACTTCCGCAGCAAGTCTCTTATGGAAGATGGTGGGGCTTTCAAGGATCGATTCAAAGCTGAGATGCTAAATTCATCCTTCTCCACTCTGAAGATTCAACCTACAGAACCCAGGGACTCAGCTGTGTATCTGTGTGCCAGCAGTTTTGGGACAGCTAGTGCAGAAACGCTGTATTTTGGCTCAGGAACCAGACTGACTGTTCTCGAGGATCTGAGAAATGTGACTCCACCCAAGGTCTCCTTGTTTGAGCCATCAAAAGCAGAGATTGCAAACAAACAAAAGGCTACCCTCGTGTGCTTGGCCAGGGGCTTCTTCCCCTGACACGTGGAGCTGAGCTGGTGGGTGAATGGCAAGGAGGTCCACAGTGGGGTCAGCACGGACCCTCAGGCCTACAAGGAGAGCAATTATAGCTACTGCCTGAGCAGCCGCCTGAGGGTCTCTGCTACCTTCTGGCACAATCCTCGAAACCACTTCCGCTGTCAAGTGCAGTTCCATGGGCTTTCAGAGGAGGACAAGTGGCCAGAGGGCTCACCCAAACCTGTCACACAGAACATCAGTGCAGAGGCCTGGGGCCGAGCAGACTGTGGAATCACTTCAGCATCCTATCATCAGGGGGTTCTGTCTGCAACCATCCTCTATGAGATCCTACTGGGGAAGGCCACCCTATATGCTGTGCTGGTCAGTGGCCTGGTGCTGATGGCCATGGTCAAGAAAAAAAATTCCTGA(SEQ ID NO: 193) MGPVTCSVLVLLLMLRRSNGDGDSVTQTEGLVTLTEGLPVMLNCTYQTIYSNPFLFWYVQHLNESPRLLLKSFTDNKRTEHQGFHATLHKSSSSFHLQKSSAQLSDSALYYCAFDTNAYKVIF (Sequence No. 194) MRVRLISAVVLCSLGTGLVDMKVTQMPRYLIKRMGENVLLECGQDMSHETMYWYRQDPGLGLQLIYISYDVDSNSEGDIPKGYRVSRKKREHFSLILDSAKTNQTSVYFCASSSTNTEVF (Sequence No. 195) MGPVTCSVLVLLLMLRRSNGDGDSVTQTEGLVTLTEGLPVMLNCTYQTIYSNPFLFWYVQHLNESPRLLLKSFTDNKRTEHQGFHATLHKSSSSFHLQKSSAQLSDSALYYCAFDTNAYKVIFGKGTHLHVLPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNTTYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLRILLLKVAGFNLLMTLRLWSSL (Sequence No. 196) MRVRLISAVVLCSLGTGLVDMKVTQMPRYLIKRMGENVLLECGQDMSHETMYWYRQDPGLGLQLIYISYDVDSNSEGDIPKGYRVSRKKREHFSLILDSAKTNQTSVYFCASSSTNTEVFFGKGTRLTVVEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVM (Sequence No. 197) MVLVTILLLSAFFSLRGNSAQSVDQPDAHVTLSEGASLELRCSYSYSAAPYLFWYVQYPGQSLQFLLKYITGDTVVKGTKGFEAEFRKSNSSFNLKKSPAHWSDSAKYFCALEGPDTGNYKYV (SEQ ID NO: 198) MGIQTLCCVIFYVLIANHTDAGVTQTPRHEVAEKGQTIILKCEPVSGHNDLFWYRQTKIQGLELLSYFRSKSLMEDGGAFKDRFKAEMLNSSFSTLKIQPTEPRDSAVYLCASSFGTASAETLY (SEQ ID NO: 199) MVLVTILLLSAFFSLRGNSAQSVDQPDAHVTLSEGASLELRCSYSYSAAPYLFWYVQYPGQSLQFLLKYITGDTVVKGTKGFEAEFRKSNSSFNLKKSPAHWSDSAKYFCALEGPDTGNYKYVFGAGTRLKVIAHIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKTVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLSVMGLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 200) MGIQTLCCVIFYVLIANHTDAGVTQTPRHEVAEKGQTIILKCEPVSGHNDLFWYRQTKIQGLELLSYFRSKSLMEDGGAFKDRFKAEMLNSSFSTLKIQPTEPRDSAVYLCASSFGTASAETLYFGSGTRLTVLEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPHVELSWWVNGKEVHSGVSTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYHQGVLSATILYEILLGKATLYAVLVSGLVLMAMVKKKNS (SEQ ID NO: 201)

[0279] In some embodiments, the TCR construct comprises a MAGE-A4-specific TCR chain. In some embodiments, a TCR construct comprising a MAGE-A4-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the epitope GVYDGREHTV (SEQ ID NO: 202). In some embodiments, a TCR construct comprising a MAGE-A4-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the epitope FMNKFIYEI (SEQ ID NO: 203). In some embodiments, the MAGE-A4-specific TCR sequences, TCR variable domain sequences, CDR sequences, and / or TCR constant domain sequences are described in International Publication Nos. WO2017 / 174824A1 and WO2021 / 229212A1, each of which is incorporated herein by reference for the purposes described herein. In certain embodiments, the anti-MAGE-A4 TCR alpha chain variable domain may have an M4V or M4L amino acid substitution. In certain embodiments, the anti-MAGE-A4 TCR beta chain variable domain may have an N10E amino acid substitution.

[0280] In some embodiments, a TCR construct comprising a MAGE-A4-specific TCR chain 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 a MAGE-A4-specific TCR chain 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. ATGAAGAAGCACCTGACCACCTTTCTCGTGATCCTGTGGCTGTACTTCTACCGGGGCAACGGCAAGAACCAGGTGGAACAGAGCCCCCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCCCCCTTCAGCAACCTGCGGTGGTACAAGCAGGACACCGGCAGAGGCCCTGTGTCCCTGACCATCCTGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTACACCGCCACCCTGGACGCCGATACAAAGCAGAGCAGCCTGCACATCACCGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGGCGGCACAGACAGCTGGGGCAAGCTGCAGTTTGGCGCCGGAACACAGGTGGTCGTGACCCCCGACATCCAGAACCCTGACCCTGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGACCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTCAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCAGAATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCCAGCGGCAGCCGGGCCAAGAGA(SEQ ID NO: 204) ATGGCCAGCCTGCTGTTCTTCTGCGGCGCCTTCTACCTGCTGGGCACCGGCTCTATGGATGCCGACGTGACCCAGACCCCCCGGAACAGAATCACCAAGACCGGCAAGCGGATCATGCTGGAATGCTCCCAGACCAAGGGCCACGACCGGATGTACTGGTACAGACAGGACCCTGGCCTGGGCCTGCGGCTGATCTACTACAGCTTCGACGTGAAGGACATCAACAAGGGCGAGATCAGCGACGGCTACAGCGTGTCCAGACAGGCTCAGGCCAAGTTCAGCCTGTCCCTGGAAAGCGCCATCCCCAACCAGACCGCCCTGTACTTTTGTGCCACAAGCGGCCAGGGCGCCTACGAGGAGCAGTTCTTTGGCCCTGGCACCCGGCTGACAGTGCTGGAAGATCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTTCTGAGGCCGAAATCAGCCACACCCAGAAAGCCACACTCGTGTGTCTGGCCACCGGCTTCTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTGTCCACCGATCCCCAGCCTCTGAAAGAACAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACCACTTCAGATGCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACAGAGCCAAGCCCGTGACACAGATCGTGTCTGCCGAAGCTTGGGGGCGCGCCGATTGTGGCTTTACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGAAAGGCCACACTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC(SEQ ID NO: 205) MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSSGSRAKR(SEQ ID NO: 206) MKKHLTTFLVILWLYFYRGNGKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQFGAGTQVVVTPD(SEQ ID NO: 207) MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG(SEQ ID NO: 208) MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFFGPGTRLTVLE(SEQ ID NO: 209) MKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTIMTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQF (SEQ ID NO: 210) MKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTIVTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQF (SEQ ID NO: 211) MKNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRGPVSLTILTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICVVSGGTDSWGKLQF (SEQ ID NO: 212) MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYNEQFF (SEQ ID NO: 213) MASLLFFCGAFYLLGTGSMDADVTQTPRNRITKTGKRIMLECSQTKGHDRMYWYRQDPGLGLRLIYYSFDVKDINKGEISDGYSVSRQAQAKFSLSLESAIPNQTALYFCATSGQGAYEEQFF (SEQ ID NO: 214)

[0281] In some embodiments, the TCR construct comprises a Wilms tumor antigen (WT1)-specific TCR chain. In some embodiments, a TCR construct comprising a WT1-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the epitope VLDFAPPGA (SEQ ID NO: 215). In some embodiments, a TCR construct comprising a WT1-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the epitope RMFPNAPYL (SEQ ID NO: 216). In some embodiments, the WT1-specific TCR sequences, TCR variable domain sequences, CDR sequences, and / or TCR constant domain sequences are described in International Publication Nos. WO2020 / 185796A1 and WO2021 / 034976A1, each of which is incorporated herein by reference for the purposes described herein. In some embodiments, the leader sequence and / or signal peptide may be removed from the TCR amino acid sequence, and the percent sequence identity may be calculated based on the TCR amino acid sequence without the leader sequence and / or signal peptide.

[0282] In some embodiments, a TCR construct comprising a WT1-specific TCR chain 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 a WT1-specific TCR chain 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. ATGGAGACACTGCTGGGACTACTGATTCTGTGGCTGCAACTGCAATGGGTGAGCAGCAAACAGGAGGTTACCCAGATTCCTGCTGCTCTGTCTGTTCCTGAAGGCGAGAATCTGGTGCTGAACTGCAGCTTCACAGATAGCGCCATCTACAACCTGCAGTGGTTCAGACAGGATCCTGGAAAAGGCCTGACAAGCCTGCTGCTGATTCAGAGCTCTCAGAGAGAGCAGACATCTGGAAGACTGAATGCTAGCCTGGACAAGTCTAGCGGCAGAAGCACCCTGTATATTGCCGCCTCTCAACCTGGAGATTCTGCCACATACCTGTGTGCTGTGAAGGAGACATCTGGCTCTAGACTGACCTTTGGCGAGGGAACACAACTGACCGTGAATCCTGAC(SEQ ID NO: 217) ATGACCAGAGTTAGCCTGTTATGGGCTGTGGTGGTGAGCACATGTCTGGAATCTGGAATGGCCCAGACAGTGACACAGTCTCAGCCTGAAATGTCTGTGCAGGAAGCCGAAACCGTTACACTGAGCTGCACCTACGATACAAGCGAGAACAACTACTACCTGTTCTGGTACAAGCAGCCCCCCTCTAGGCAGATGATCCTGGTGATCAGACAGGAGGCCTATAAACAGCAGAATGCCACAGAGAACCGGTTCAGCGTGAACTTCCAGAAAGCCGCCAAGAGCTTCAGCCTGAAGATCTCTGATTCTCAGCTGGGCGATACAGCCATGTACTTTTGCGCCTTCATCTACCCCAGCTACACAAGCGGCACATACAAGTACATCTTCGGCACCGGCACAAGACTGAAGGTTCTGGCCAAC(SEQ ID NO: 218) ATGGCCATGTTACTAGGAGCGAGCGTGCTGATTCTGTGGTTACAGCCTGATTGGGTGAACTCTCAGCAGAAGAACGATGATCAGCAGGTGAAGCAGAACAGCCCCTCTCTGTCTGTGCAGGAAGGCAGAATCAGCATCCTGAATTGCGATTACACCAACAGCATGTTCGACTACTTCCTGTGGTACAAGAAGTACCCCGCCGAGGGCCCTACCTTTCTGATCAGCATCTCTAGCATCAAGGACAAGAACGAAGATGGCAGATTCACCGTGTTCCTGAACAAGAGCGCCAAGCACCTGAGCCTGCACATTGTGCCTTCTCAACCTGGAGATTCTGCCGTGTACTTTTGTGCTGCCTCTGGAACAGGCGGAAGCTATATCCCCACATTTGGAAGAGGAACAAGCCTGATCGTGCACCCTTAC (SEQ ID NO: 219) ATGGCCATGTTACTAGGAGCGAGCGTGCTGATTCTGTGGTTACAGCCTGATTGGGTGAACTCTCAGCAGAAGAACGATGATCAGCAGGTGAAGCAGAACAGCCCCTCTCTGTCTGTGCAGGAAGGCAGAATCAGCATCCTGAATTGCGATTACACCAACAGCATGTTCGACTACTTCCTGTGGTACAAGAAGTACCCCGCCGAGGGCCCTACCTTTCTGATCAGCATCTCTAGCATCAAGGACAAGAACGAAGATGGCAGATTCACCGTGTTCCTGAACAAGAGCGCCAAGCACCTGAGCCTGCACATTGTGCCTTCTCAACCTGGAGATTCTGCCGTGTACTTTTGTGCTGCCTCTGGCATTGGCGACTACAAACTGAGCTTTGGAGCCGGCACAACAGTGACCGTTAGAGCCAAT (SEQ ID NO: 220) ATGGTGAAGATCCGGCAGTTCCTCCTGGCTATTCTGTGGCTGCAACTGTCTTGTGTGTCTGCTGCCAAGAATGAAGTGGAGCAGTCTCCCCAGAACCTTACAGCCCAGGAAGGCGAGTTTATCACCATCAACTGCAGCTATTCTGTGGGCATTAGCGCCCTGCATTGGCTGCAGCAACACCCTGGAGGAGGAATTGTGTCTCTGTTTATGCTGTCTTCTGGCAAGAAGAAGCACGGCCGGCTGATTGCCACCATCAACATCCAGGAGAAGCACTCTTCTCTGCACATTACAGCCTCTCATCCCAGGGATTCTGCCGTGTACATCTGTGCCGTGAGAACCAGCTACGATAAGGTGATTTTCGGACCAGGCACCTCTCTGAGCGTGATCCCCAAT(SEQ ID NO: 221) ATGAAGAGCCTGAGAGTCCTGCTGGTGATTTTGTGGCTGCAGCTGTCTTGGGTTTGGTCTCAGCAGAAAGAAGTGGAGCAGAATAGCGGCCCTCTGTCTGTTCCTGAAGGCGCTATTGCTAGCCTGAATTGCACATACAGCGATAGAGGATCTCAGAGCTTCTTCTGGTACCGGCAGTACAGCGGCAAGAGCCCAGAACTGATCATGTTCATCTACAGCAATGGCGACAAGGAGGATGGCAGGTTTACAGCCCAGCTGAACAAGGCCAGCCAGTATGTTTCTCTGCTGATCAGAGATAGCCAGCCTAGCGATTCTGCCACCTACCTGTGTGCCGTGAACTTACTTGGAGCTACAGGATACTCTACACTGACCTTCGGCAAAGGCACCATGCTGCTGGTGAGCCCTGAT(SEQ ID NO: 222) ATGTGGGGCGTTTTCCTTCTGTATGTGAGCATGAAGATGGGCGGCACAACAGGCCAGAACATCGATCAGCCTACCGAGATGACAGCCACAGAAGGAGCTATTGTTCAGATCAACTGCACCTACCAGACAAGCGGCTTCAACGGCCTGTTCTGGTACCAGCAGCATGCTGGAGAAGCTCCTACATTTCTGAGCTACAATGTGCTGGATGGCCTGGAGGAGAAAGGCAGGTTTAGCAGCTTCCTGAGCAGGTCTAAGGGCTATTCTTATCTGCTGCTGAAGGAGCTGCAGATGAAGGATTCCGCCAGCTACCTGTGTGCCGTTAGGGGCATCAATGATTACAAGCTGAGCTTTGGAGCCGGAACAACAGTGACCGTGAGAGCCAAC(SEQ ID NO: 223) ATGGAGAAGATGCTGGAGTGTGCGTTCATCGTTCTGTGGCTGCAACTTGGATGGCTGTCTGGAGAGGATCAGGTTACACAGTCTCCTGAAGCCCTGAGACTGCAAGAAGGAGAAAGCTCTAGCCTGAACTGCAGCTACACAGTGTCTGGACTGAGAGGCCTGTTCTGGTACAGACAGGATCCTGGAAAAGGCCCAGAGTTCCTGTTTACCCTGTATTCTGCCGGCGAGGAGAAGGAGAAAGAGAGACTGAAAGCTACCCTGACCAAGAAGGAGAGCTTCCTGCACATTACCGCCCCCAAACCTGAGGATTCTGCCACATATCTGTGTGCCGTGATTACCGGCTTTCAGAAGCTGGTGTTTGGCACAGGCACCAGACTGCTGGTTTCTCCCAAT(SEQ ID NO: 224) ATGAGACTGGTGGCACGCGTAACTGTGTTTCTGACCTTTGGCACCATCATCGATGCCAAGACAACCCAGCCTACAAGCATGGACTGTGCCGAGGGAAGAGCTGCTAATCTGCCATGTAATCACAGCACAATCAGCGGCAACGAGTACGTGTACTGGTACCGGCAGATCCACTCTCAAGGACCTCAGTACATCATTCATGGCCTGAAGAACAACGAGACCAACGAGATGGCCAGCCTGATCATCACCGAGGACAGGAAGTCTTCTACCCTGATTCTGCCTCATGCTACACTGAGAGATACCGCCGTGTACTACTGCATTGCCGGAGTGGGAAGAGGCCAGAATTTCGTGTTTGGACCTGGAACAAGACTGAGCGTTCTGCCCTAT(SEQ ID NO: 225) ATGGAGAAGAACCCCTTGGCAGCACCTCTGCTTATTCTGTGGTTCCACCTGGATTGTGTGAGCAGCATCCTGAATGTGGAGCAGTCTCCTCAGAGCCTGCATGTGCAAGAAGGCGATAGCACCAATTTCACCTGCAGCTTTCCAAGCAGCAACTTCTACGCCCTGCACTGGTACAGATGGGAAACCGCCAAATCTCCTGAAGCCCTGTTTGTGATGACCCTGAATGGCGACGAGAAGAAGAAGGGCAGAATTAGCGCCACCCTGAATACCAAGGAGGGCTACAGCTACCTGTACATCAAGGGCTCTCAACCTGAGGATTCTGCCACCTACCTTTGCGCCTTTCACCCCAATTTCGGCAACGAGAAACTGACCTTTGGAACCGGAACAAGGCTGACCATCATCCCCAAC(SEQ ID NO: 226) ATGGAGAAGATGCTGGAGTGTGCGTTCATCGTTCTGTGGCTGCAACTTGGATGGCTGTCTGGAGAGGATCAGGTTACACAGTCTCCTGAAGCCCTGAGACTGCAAGAAGGAGAAAGCTCTAGCCTGAACTGCAGCTACACAGTGTCTGGACTGAGAGGCCTGTTCTGGTACAGACAGGATCCTGGAAAAGGCCCAGAGTTCCTGTTTACCCTGTATTCTGCCGGCGAGGAGAAGGAGAAAGAGAGACTGAAAGCTACCCTGACCAAGAAGGAGAGCTTCCTGCACATTACCGCCCCCAAACCTGAGGATTCTGCCACATATCTGTGTGCTGTTCAGCCTAGAGGAGATGGCTCTAGCAATACCGGCAAGCTGATCTTTGGCCAGGGAACAACACTGCAGGTGAAGCCTGAT(SEQ ID NO: 227) ATCCAGAATCCCGATCCTGCTGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 228) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCACATCTCTTCTCTGTTGGGTGGTTCTGGGCTTTCTGGGCACAGATCATACAGGAGCTGGAGTTAGCCAGTCTCCTAGGTATAAGGTGACCAAGAGGGGACAGGATGTGGCTCTGAGATGTGACCCTATTAGCGGACATGTGAGCCTGTACTGGTACAGACAAGCTCTGGGACAAGGACCCGAGTTTCTGACCTACTTCAACTATGAGGCCCAGCAGGACAAATCTGGACTGCCCAACGACAGATTCAGCGCCGAAAGACCAGAAGGCTCTATTAGCACACTGACCATCCAGAGAACAGAGCAGAGGGATTCTGCCATGTACAGATGCGCCAGCAGCTTAACAGGCTCTTACGAGCAGTACTTTGGACCTGGCACAAGACTGACAGTGACAGAG (SEQ ID NO: 229) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGCTGCTTCTTCTCCTCCTTCTCGGACCTGCTGGATCTGGATTAGGAGCTGTTGTGTCTCAGCACCCTTCTTGGGTGATCTGTAAAAGCGGCACAAGCGTGAAGATCGAGTGCAGAAGCCTGGACTTTCAGGCCACAACCATGTTCTGGTATAGGCAGTTCCCCAAGCAGTCTCTGATGCTGATGGCCACCTCTAATGAGGGCTCTAAGGCCACATATGAACAGGGAGTGGAGAAGGACAAGTTCCTGATCAACCACGCCTCTCTGACCCTGTCTACCCTGACAGTTACATCTGCCCACCCTGAGGATAGCAGCTTTTACATCTGTAGCGCCACACCTGAAGCCTCTAGCCCATATGAGCAGTACTTTGGCCCTGGCACCAGATTAACAGTGACAGAG (SEQ ID NO: 230) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGACCTGGACTGCTTCATTGGATGGCTCTGTGTTTGCTGGGAACAGGACATGGAGATGCTATGGTGATCCAGAACCCCAGGTATCAGGTGACCCAGTTTGGCAAACCAGTGACACTGAGCTGTTCTCAGACCCTGAACCACAACGTGATGTACTGGTACCAGCAGAAGTCTTCTCAGGCCCCTAAGCTGCTGTTCCACTACTACGACAAGGACTTCAACAACGAGGCCGATACCCCTGACAATTTCCAGAGCAGGAGGCCCAATACCAGCTTCTGTTTCCTGGACATTAGAAGCCCTGGACTGGGAGATGCTGCCATGTACCTGTGTGCCACCAGCAATTTACAGGGAAGACAACCTCAGCACTTTGGCGATGGCACAAGGCTGTCTATCCTGGAG(SEQ ID NO: 231) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGCTGAGCCCTGATCTCCCTGATTCTGCCTGGAATACCAGACTGCTGTGTCATGTGATGCTGTGTCTGCTTGGAGCCGTTTCTGTGGCTGCTGGCGTGATTCAATCTCCTAGACACCTGATCAAGGAGAAGAGAGAAACAGCCACCCTGAAGTGCTACCCCATCCCCAGACACGATACAGTGTACTGGTATCAGCAAGGACCTGGACAAGATCCCCAGTTCCTGATCAGCTTCTACGAGAAGATGCAGAGCGACAAAGGCAGCATCCCAGACAGATTTAGCGCCCAGCAGTTTAGCGACTATCACTCTGAGCTGAACATGAGCAGCCTGGAACTGGGCGATTCTGCTCTGTACTTCTGTGCCTCTTCTCTGAGACTGGGAAGAGAAACCCAGTACTTTGGACCCGGCACAAGACTGCTGGTTCTTGAG(SEQ ID NO: 232) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCACAAGACTTCTCTGCTGGGTGGTGCTTGGATTTCTGGGCACAGATCATACAGGAGCTGGAGTTAGCCAGTCTCCTAGGTACAAAGTGGCCAAGAGAGGACAGGATGTGGCTCTGAGATGTGACCCTATTAGCGGACATGTGAGCCTGTTTTGGTACCAGCAAGCTCTGGGACAAGGACCCGAGTTTCTGACCTACTTCCAGAATGAAGCCCAGCTGGATAAATCTGGACTGCCTAGCGACCGGTTCTTCGCCGAAAGACCTGAAGGATCTGTTAGCACCCTGAAGATTCAGAGAACACAGCAGGAGGACTCTGCCGTGTACCTGTGTGCCTCTTCTTTAGGACAGGCCTATGAGCAGTATTTTGGACCTGGCACCAGACTGACCGTGACAGAG(SEQ ID NO: 233) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCACAAGACTTCTCTGCTGGGTGGCCTTTTGTCTGCTGGTGGAAGAGCTGATTGAAGCTGGAGTTGTGCAGTCTCCTAGGTACAAGATCATCGAGAAGAAGCAGCCCGTGGCCTTCTGGTGTAATCCCATTTCTGGCCACAACACCCTGTACTGGTATCTGCAGAATCTGGGACAGGGCCCTGAACTGCTGATCAGATACGAGAACGAAGAAGCCGTGGACGATTCTCAACTGCCTAAGGACCGCTTTTCTGCCGAGAGGCTGAAAGGAGTGGATTCTACCCTGAAGATCCAACCTGCTGAACTGGGCGATTCTGCTGTGTACCTGTGCGCTTCTAGCCTGACAAGAGGAGCTGAAGCCTTTTTTGGACAGGGCACAAGACTGACAGTGGTGGAG(SEQ ID NO: 234) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGACCTCAGCTTCTTGGATACGTTGTGCTGTGTCTGCTTGGAGCTGGACCTCTTGAAGCTCAGGTTACCCAGAACCCCAGATACCTGATTACCGTGACAGGCAAAAAGCTGACCGTGACATGTAGCCAGAACATGAACCACGAGTACATGAGCTGGTACCGGCAGGATCCTGGATTAGGCCTGAGACAGATCTACTACAGCATGAACGTGGAGGTGACCGATAAAGGCGACGTGCCTGAGGGATACAAGGTGAGCAGAAAGGAGAAGAGGAATTTCCCCCTGATCCTGGAAAGCCCAAGCCCCAATCAGACAAGCCTGTACTTTTGTGCCAGCAGCTTTTCTGGCGGCACATATGAGCAGTACTTCGGCCCTGGCACAAGACTGACAGTTACAGAG(SEQ ID NO: 235) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGCTGAGCCCTGATCTCCCTGATTCTGCCTGGAATACCAGACTGCTGTGTCATGTGATGCTGTGTCTGCTTGGAGCCGTTTCTGTGGCTGCTGGCGTGATTCAATCTCCTAGACACCTGATCAAGGAGAAGAGAGAAACAGCCACCCTGAAGTGCTACCCCATCCCCAGACACGATACAGTGTACTGGTATCAGCAAGGACCTGGACAAGATCCCCAGTTCCTGATCAGCTTCTACGAGAAGATGCAGAGCGACAAAGGCAGCATCCCAGACAGATTTAGCGCCCAGCAGTTTAGCGACTATCACTCTGAGCTGAACATGAGCAGCCTGGAACTGGGCGATTCTGCTCTGTACTTCTGTGCCAGCAGCTATAGAGGAGGCAGCACATATGAGCAGTACTTTGGCCCTGGCACAAGACTGACAGTGACAGAG(SEQ ID NO: 236) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGAGCACCAGACTCCTTTGCTGGATGGCTTTGTGTCTGCTTGGAGCTGAGCTGTCTGAAGCTGAAGTTGCCCAGTCTCCCAGATACAAGATCACCGAGAAATCTCAGGCTGTGGCCTTCTGGTGTGACCCTATTTCTGGACACGCCACCCTGTACTGGTATAGGCAAATTCTGGGACAAGGCCCTGAACTGCTGGTGCAATTTCAGGATGAGAGCGTGGTGGACGATTCTCAACTGCCTAAGGACAGGTTTTCTGCCGAGCGGCTGAAAGGAGTTGATAGCACCCTGAAGATCCAACCTGCTGAACTGGGCGATTCTGCTATGTACCTGTGCGCCTCTTCTCAGAGAGATAGCCCTAACGAGAAGCTGTTCTTTGGCTCTGGAACCCAGCTGTCTGTGCTGGAG(SEQ ID NO: 237) CTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCACCATGGGCTGTAGACTGTTGTGTTGTGCTGTGCTGTGTCTGTTGGGAGCTGTGCCTATGGAAACAGGCGTTACCCAGACACCTAGACATCTGGTTATGGGCATGACCAACAAGAAGAGCCTGAAGTGCGAGCAGCATCTGGGCCATAACGCCATGTACTGGTATAAGCAGAGCGCCAAGAAACCACTGGAACTGATGTTCGTGTACAGCCTGGAGGAGAGGGTGGAGAATAATAGCGTGCCCAGCAGATTTAGCCCTGAGTGCCCAAATTCTTCTCACCTGTTCCTGCACCTGCACACATTACAGCCCGAGGATTCTGCCCTGTACCTGTGTGCTTCTTCTCAAGACCCTTACAAGCTGAGCGGCAATACCATCTACTTCGGCGAAGGCTCTTGGCTGACAGTGGTTGAA(SEQ ID NO: 238) GATCTGAACAAGGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTTCTGAGGCCGAGATCTCCCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTTCCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACTCCGGCGTGTGCACCGATCCCCAGCCTCTGAAAGAACAGCCCGCCCTGAACGACAGCCGGTACTGCCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACAGAGCCAAGCCCGTGACACAGATCGTGTCTGCCGAAGCCTGGGGCAGAGCCGATTGCGGCTTTACCTCCGTGTCCTATCAGCAGGGCGTGCTGAGCGCCACAATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCTGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACTTC(SEQ ID NO: 239) GACCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC(SEQ ID NO: 240) ATGAAATCCTTGAGAGTTTTACTAGTGATCCTGTGGCTTCAGTTGAGCTGGGTTTGGAGCCAACAGAAGGAGGTGGAGCAGAATTCTGGACCCCTCAGTGTTCCAGAGGGAGCCATTGCCTCTCTCAACTGCACTTACAGTGACCGAGGTTCCCAGTCCTTCTTCTGGTACAGACAATATTCTGGGAAAAGCCCTGAGTTGATAATGTTCATATACTCCAATGGTGACAAAGAAGATGGAAGGTTTACAGCACAGCTCAATAAAGCCAGCCAGTATGTTTCTCTGCTCATCAGAGACTCCCAGCCCAGTGATTCAGCCACCTACCTCTGTGCCGTGAACATAGGAAACCATGACATGCGCTTTGGAGCAGGGACCAGACTGACAGTAAAACCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 241) ATGGAGAAAATGTTGGAGTGTGCATTCATAGTCTTGTGGCTTCAGCTTGGCTGGTTGAGTGGAGAAGACCAGGTGACGCAGAGTCCCGAGGCCCTGAGACTCCAGGAGGGAGAGAGTAGCAGTCTCAACTGCAGTTACACAGTCAGCGGTTTAAGAGGGCTGTTCTGGTATAGGCAAGATCCTGGGAAAGGCCCTGAATTCCTCTTCACCCTGTATTCAGCTGGGGAAGAAAAGGAGAAAGAAAGGCTAAAAGCCACATTAACAAAGAAGGAAAGCTTTCTGCACATCACAGCCCCTAAACCTGAAGACTCAGCCACTTATCTCTGTGCTGTGCAGACCATGGACGGTAACCAGTTCTATTTTGGGACAGGGACAAGTTTGACGGTCATTCCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 242) ATGGCATGCCCTGGCTTCCTGTGGGCACTTGTGATCTCCACCTGTCTTGAATTTAGCATGGCTCAGACAGTCACTCAGTCTCAACCAGAGATGTCTGTGCAGGAGGCAGAGACCGTGACCCTGAGCTGCACATATGACACCAGTGAGAGTGATTATTATTTATTCTGGTACAAGCAGCCTCCCAGCAGGCAGATGATTCTCGTTATTCGCCAAGAAGCTTATAAGCAACAGAATGCAACAGAGAATCGTTTCTCTGTGAACTTCCAGAAAGCAGCCAAATCCTTCAGTCTCAAGATCTCAGACTCACAGCTGGGGGATGCCGCGATGTATTTCTGTGCTTCCAGTCCAGGAACCTACAAATACATCTTTGGAACAGGCACCAGGCTGAAGGTTTTAGCAAATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 243) ATGACACGAGTTAGCTTGCTGTGGGCAGTCGTGGTCTCCACCTGTCTTGAATCCGGCATGGCCCAGACAGTCACTCAGTCTCAACCAGAGATGTCTGTGCAGGAGGCAGAGACTGTGACCCTGAGTTGCACATATGACACCAGTGAGAGTAATTATTATTTGTTCTGGTACAAACAGCCTCCCAGCAGGCAGATGATTCTCGTTATTCGCCAAGAAGCTTATAAGCAACAGAATGCAACGGAGAATCGTTTCTCTGTGAACTTCCAGAAAGCAGCCAAATCCTTCAGTCTCAAGATCTCAGACTCACAGCTGGGGGACACTGCGATGTATTTCTGTGCTTTCAACCCTTGGGAGAACTATGGTCAGAATTTTGTCTTTGGTCCCGGAACCAGATTGTCCGTGCTGCCCTATATCCAGAACCCTGACCCTGCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTTCTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAACAGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 244) ATGAAGAGCCTGAGAGTCCTGCTGGTGATTTTGTGGCTGCAGCTGTCTTGGGTTTGGTCTCAGCAGAAAGAAGTGGAGCAGAATAGCGGCCCTCTGTCTGTTCCTGAAGGCGCTATTGCTAGCCTGAATTGCACATACAGCGATAGAGGATCTCAGAGCTTCTTCTGGTACCGGCAGTACAGCGGCAAGAGCCCAGAACTGATCATGTTCATCTACAGCAATGGCGACAAGGAGGATGGCAGGTTTACAGCCCAGCTGAACAAGGCCAGCCAGTATGTTTCTCTGCTGATCAGAGATAGCCAGCCTAGCGATTCTGCCACCTACCTGTGTGCCGTGAACATCGGAAATCACGACATGAGATTTGGAGCCGGCACAAGACTGACCGTGAAGCCCAATATCCAGAACCCTGATCCTGCTGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 245) ATGGAGAAGATGCTGGAGTGTGCGTTCATCGTTCTGTGGCTGCAACTTGGATGGCTGTCTGGAGAGGATCAGGTTACACAGTCTCCTGAAGCCCTGAGACTGCAAGAAGGAGAAAGCTCTAGCCTGAACTGCAGCTACACAGTGTCTGGACTGAGAGGCCTGTTCTGGTACAGACAGGATCCTGGAAAAGGCCCAGAGTTCCTGTTTACCCTGTATTCTGCCGGCGAGGAGAAGGAGAAAGAGAGACTGAAAGCTACCCTGACCAAGAAGGAGAGCTTCCTGCACATTACCGCCCCCAAACCTGAGGATTCTGCCACATATCTGTGTGCTGTGCAGACCATGGATGGCAACCAGTTCTACTTCGGCACAGGCACATCTCTGACCGTTATCCCCAATATCCAGAACCCTGATCCTGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 246) ATGGCTTGTCCTGGATTCTTATGGGCTCTGGTGATCAGCACCTGTCTGGAGTTCTCTATGGCCCAGACAGTGACACAGTCTCAGCCTGAAATGTCTGTGCAGGAAGCCGAAACCGTGACACTGTCTTGCACCTACGATACAAGCGAGAGCGACTACTACCTGTTCTGGTACAAGCAGCCTCCCTCTAGGCAGATGATCCTGGTGATTAGACAGGAGGCCTACAAACAGCAGAATGCCACCGAGAACCGGTTTAGCGTGAACTTCCAGAAAGCCGCCAAGAGCTTCAGCCTGAAAATCTCTGACAGCCAGCTGGGAGATGCTGCCATGTACTTTTGTGCCAGCTCTCCAGGCACCTACAAGTACATTTTTGGCACCGGCACCAGACTGAAGGTGCTGGCCAATATCCAGAATCCCGATCCTGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 247) ATGACCAGAGTTAGCCTGTTATGGGCTGTGGTGGTGAGCACATGTCTGGAATCTGGAATGGCCCAGACAGTGACACAGTCTCAGCCTGAAATGTCTGTGCAGGAAGCCGAAACCGTTACACTGAGCTGCACCTACGATACAAGCGAGAGCAACTACTACCTGTTCTGGTACAAGCAGCCCCCTTCTAGGCAGATGATCCTGGTGATCAGACAGGAGGCCTATAAACAGCAGAATGCCACCGAGAACCGGTTTAGCGTGAACTTCCAGAAAGCCGCCAAGAGCTTCAGCCTGAAAATCTCTGACAGCCAGCTGGGCGATACAGCCATGTACTTTTGTGCCTTCAACCCCTGGGAGAACTATGGCCAGAATTTCGTGTTCGGCCCTGGCACCAGACTGTCTGTTCTGCCTTATATCCAGAACCCCGATCCTGCTGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTCAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAGGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGCTGA(SEQ ID NO: 248) ATGGGCTGCAGGCTGCTCTGCTGTGCGGTTCTCTGTCTCCTGGGAGCAGTTCCCATAGACACTGAAGTTACCCAGACACCAAAACACCTGGTCATGGGAATGACAAATAAGAAGTCTTTGAAATGTGAACAACATATGGGGCACAGGGCTATGTATTGGTACAAGCAGAAAGCTAAGAAGCCACCGGAGCTCATGTTTGTCTACAGCTATGAGAAACTCTCTATAAATGAAAGTGTGCCAAGTCGCTTCTCACCTGAATGCCCCAACAGCTCTCTCTTAAACCTTCACCTACACGCCCTGCAGCCAGAAGACTCAGCCCTGTATCTCTGCGCCAGCAGCCAAGGGACTAGCGGGGCAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCTGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCACCTCCGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGGCCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAGGATTCCAGAGGCTAG(SEQ ID NO: 249) ATGAGCATCGGCCTCCTGTGCTGTGCAGCCTTGTCTCTCCTGTGGGCAGGTCCAGTGAATGCTGGTGTCACTCAGACCCCAAAATTCCAGGTCCTGAAGACAGGACAGAGCATGACACTGCAGTGTGCCCAGGATATGAACCATGAATACATGTCCTGGTATCGACAAGACCCAGGCATGGGGCTGAGGCTGATTCATTACTCAGTTGGTGCTGGTATCACTGACCAAGGAGAAGTCCCCAATGGCTACAATGTCTCCAGATCAACCACAGAGGATTTCCCGCTCAGGCTGCTGTCGGCTGCTCCCTCCCAGACATCTGTGTACTTCTGTGCCAGCAGTTACTCTCTTTGGGACCTTCAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCTGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCACCTCCGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGGCCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAGGATTCCAGAGGCTAG(SEQ ID NO: 250) ATGGGCACCAGCCTCCTCTGCTGGATGGCCCTGTGTCTCCTGGGGGCAGATCACGCAGATACTGGAGTCTCCCAGGACCCCAGACACAAGATCACAAAGAGGGGACAGAATGTAACTTTCAGGTGTGATCCAATTTCTGAACACAACCGCCTTTATTGGTACCGACAGACCCTGGGGCAGGGCCCAGAGTTTCTGACTTACTTCCAGAATGAAGCTCAACTAGAAAAATCAAGGCTGCTCAGTGATCGGTTCTCTGCAGAGAGGCCTAAGGGATCTTTCTCCACCTTGGAGATCCAGCGCACAGAGCAGGGGGACTCGGCCATGTATCTCTGTGCCAGCAGCTTTTCAGACGGGGGGGCTACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCTGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCACCTCCGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGGCCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAGGATTCCAGAGGCTAG(SEQ ID NO: 251) ATGCTGCTGCTTCTGCTGCTTCTGGGGCCAGCAGGCTCCGGGCTTGGTGCTGTCGTCTCTCAACATCCGAGCTGGGTTATCTGTAAGAGTGGAACCTCTGTGAAGATCGAGTGCCGTTCCCTGGACTTTCAGGCCACAACTATGTTTTGGTATCGTCAGTTCCCGAAACAGAGTCTCATGCTGATGGCAACTTCCAATGAGGGCTCCAAGGCCACATACGAGCAAGGCGTCGAGAAGGACAAGTTTCTCATCAACCATGCAAGCCTGACCTTGTCCACTCTGACAGTGACCAGTGCCCATCCTGAAGACAGCAGCTTCTACATCTGCAGTGCTAGACCCCATTCTCTCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTGAAAAACGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCCCACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTACCCCGACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCTGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTCACCTCCGAGTCTTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGGCCACCTTGTATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTCAAGAGAAAGGATTCCAGAGGCTAG(SEQ ID NO: 252) ATGGGCTGTAGACTGTTGTGTTGTGCTGTGCTGTGTCTGTTGGGAGCTGTGCCTATCGATACAGAGGTGACCCAGACCCCTAAACATCTGGTTATGGGCATGACCAACAAGAAGAGCCTGAAGTGCGAGCAGCACATGGGCCATAGGGCCATGTATTGGTATAAGCAGAAGGCCAAGAAACCTCCTGAGCTGATGTTCGTGTACAGCTACGAGAAGCTGAGCATCAACGAGAGCGTGCCCAGCAGATTTTCTCCTGAGTGCCCTAATTCTAGCCTGCTGAATCTGCACCTGCATGCTCTGCAGCCTGAGGATTCTGCTCTGTACCTGTGTGCTTCTTCTCAGGGCACATCTGGAGCTGATACACAGTACTTCGGACCTGGCACAAGACTGACAGTGCTGGAAGACCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC(SEQ ID NO: 253) ATGTCTATCGGTCTGCTGTGCTGTGCTGCTCTTTCTCTGCTTTGGGCTGGACCTGTGAATGCTGGAGTTACACAAACCCCCAAGTTCCAAGTGCTGAAGACAGGACAGAGCATGACCCTGCAGTGTGCTCAGGACATGAATCACGAGTACATGAGCTGGTACAGACAGGATCCTGGAATGGGCCTGAGGCTGATCCACTACTCTGTTGGAGCCGGAATTACAGATCAGGGAGAAGTGCCAAATGGCTACAACGTGAGCAGGAGCACAACCGAGGACTTCCCCTTAAGACTGTTGTCTGCTGCTCCATCTCAGACAAGCGTGTACTTTTGCGCCAGCTCCTACTCTCTGTGGGATCTGCAGGAAACCCAGTACTTTGGACCAGGCACAAGACTGTTAGTGCTGGAGGACCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC(SEQ ID NO: 254) ATGGGCACATCTCTTCTCTGCTGGATGGCTCTTTGTCTGCTTGGAGCCGATCATGCCGATACAGGAGTTAGCCAGGATCCTAGACACAAGATCACCAAGAGAGGCCAGAATGTGACCTTCCGGTGCGATCCTATCTCTGAGCACAACAGGCTGTACTGGTACAGACAAACACTGGGACAAGGACCTGAGTTCCTGACCTACTTCCAGAACGAAGCCCAGCTGGAGAAGTCTAGACTTCTGAGCGACAGATTTAGCGCCGAGAGACCTAAAGGCAGCTTTAGCACCCTGGAGATCCAGAGAACAGAACAGGGCGATTCTGCCATGTACCTGTGTGCTAGCAGCTTTTCTGATGGAGGCGCCACCGATACACAGTATTTCGGACCTGGCACAAGACTGACAGTGCTGGAGGACCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC(SEQ ID NO: 255) ATGCTGCTTCTTCTCCTCCTTCTCGGACCTGCTGGATCTGGATTAGGAGCTGTTGTGTCTCAGCACCCTTCTTGGGTGATCTGTAAAAGCGGCACAAGCGTGAAGATCGAGTGCAGAAGCCTGGACTTTCAGGCCACAACCATGTTCTGGTATAGGCAGTTCCCCAAGCAGTCTCTGATGCTGATGGCCACCTCTAATGAGGGCTCTAAGGCCACATATGAACAGGGAGTGGAGAAGGACAAGTTCCTGATCAACCACGCCTCTCTGACCCTGTCTACCCTGACAGTTACATCTGCCCACCCTGAGGATAGCAGCTTTTACATCTGTAGCGCCAGACCTCACAGCCTGACCGATACACAGTACTTTGGCCCTGGCACAAGACTGACAGTGTTAGAAGACCTGAAGAACGTGTTCCCCCCAGAGGTGGCCGTGTTCGAGCCTAGCGAGGCCGAGATCAGCCACACCCAGAAAGCCACCCTCGTGTGCCTGGCCACCGGCTTTTACCCCGACCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTCTGCACCGACCCCCAGCCCCTGAAAGAGCAGCCCGCCCTGAACGACAGCCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCCGGAACCACTTCAGATGCCAGGTGCAGTTCTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGACCGGGCCAAGCCCGTGACCCAGATCGTGTCTGCTGAGGCCTGGGGCAGAGCCGATTGCGGCTTCACCAGCGAGAGCTACCAGCAGGGCGTGCTGAGCGCCACCATCCTGTACGAGATCCTGCTGGGCAAGGCCACCCTGTACGCCGTGCTGGTGTCCGCCCTGGTGCTGATGGCCATGGTCAAGCGGAAGGACAGCCGGGGC(SEQ ID NO: 256) METLLGLLILWLQLQWVSSKQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVKETSGSRLTFGEGTQLTVNP (SEQ ID NO: 257) MTRVSLLWAVVVSTCLESGMAQTVTQSQPEMSVQEAETVTLSCTYDTSENNYYLFWYKQPPSRQMILVIRQEAYKQQNATENRFSVNFQKAAKSFSLKISDSQLGDTAMYFCAFIYPSYTSGTYKYIFGTGTRLKVLAN (SEQ ID NO: 258) MAMLLGASVLILWLQPDWVNSQQKNDDQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAASGTGGSYIPTFGRGTSLIVHPY (SEQ ID NO: 259) MAMLLGASVLILWLQPDWVNSQQKNDDQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAASGIGDYKLSFGAGTTVTVRAN (SEQ ID NO: 260) MVKIRQFLLAILWLQLSCVSAAKNEVEQSPQNLTAQEGEFITINCSYSVGISALHWLQQHPGGGIVSLFMLSSGKKKHGRLIATINIQEKHSSLHITASHPRDSAVYICAVRTSYDKVIFGPGTSLSVIPN (SEQ ID NO: 261) MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVNLLGATGYSTLTFGKGTMLLVSP (SEQ ID NO: 262) MWGVFLLYVSMKMGGTTGQNIDQPTEMTATEGAIVQINCTYQTSGFNGLFWYQQHAGEAPTFLSYNVLDGLEEKGRFSSFLSRSKGYSYLLLKELQMKDSASYLCAVRGINDYKLSFGAGTTVTVRAN (SEQ ID NO: 263) MEKMLECAFIVLWLQLGWLSGEDQVTQSPEALRLQEGESSSLNCSYTVSGLRGLFWYRQDPGKGPEFLFTLYSAGEEKEKERLKATLTKKESFLHITAPKPEDSATYLCAVITGFQKLVFGTGTRLLVSPN (SEQ ID NO: 264) MRLVARVTVFLTFGTIIDAKTTQPTSMDCAEGRAANLPCNHSTISGNEYVYWYRQIHSQGPQYIIHGLKNNETNEMASLIITEDRKSSTLILPHATLRDTAVYYCIAGVGRGQNFVFGPGTRLSVLPY (SEQ ID NO: 265) MEKNPLAAPLLILWFHLDCVSSILNVEQSPQSLHVQEGDSTNFTCSFPSSNFYALHWYRWETAKSPEALFVMTLNGDEKKKGRISATLNTKEGYSYLYIKGSQPEDSATYLCAFHPNFGNEKLTFGTGTRLTIIPN (SEQ ID NO: 266) MEKMLECAFIVLWLQLGWLSGEDQVTQSPEALRLQEGESSSLNCSYTVSGLRGLFWYRQDPGKGPEFLFTLYSAGEEKEKERLKATLTKKESFLHITAPKPEDSATYLCAVQPRGDGSSNTGKLIFGQGTTLQVKP (SEQ ID NO: 267) IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 268) MGTSLLCWVVLGFLGTDHTGAGVSQSPRYKVTKRGQDVALRCDPISGHVSLYWYRQALGQGPEFLTYFNYEAQQDKSGLPNDRFSAERPEGSISTLTIQRTEQRDSAMYRCASSLTGSYEQYFGPGTRLTVTE (SEQ ID NO: 269) MLLLLLLLGPAGSGLGAVVSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLMATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSATPEASSPYEQYFGPGTRLTVTE (SEQ ID NO: 270) MGPGLLHWMALCLLGTGHGDAMVIQNPRYQVTQFGKPVTLSCSQTLNHNVMYWYQQKSSQAPKLLFHYYDKDFNNEADTPDNFQSRRPNTSFCFLDIRSPGLGDAAMYLCATSNLQGRQPQHFGDGTRLSILE (SEQ ID NO: 271) MLSPDLPDSAWNTRLLCHVMLCLLGAVSVAAGVIQSPRHLIKEKRETATLKCYPIPRHDTVYWYQQGPGQDPQFLISFYEKMQSDKGSIPDRFSAQQFSDYHSELNMSSLELGDSALYFCASSLRLGRETQYFGPGTRLLVLE (SEQ ID NO: 272) MGTRLLCWVVLGFLGTDHTGAGVSQSPRYKVAKRGQDVALRCDPISGHVSLFWYQQALGQGPEFLTYFQNEAQLDKSGLPSDRFFAERPEGSVSTLKIQRTQQEDSAVYLCASSLGQAYEQYFGPGTRLTVTE (SEQ ID NO: 273) MGTRLLCWVAFCLLVEELIEAGVVQSPRYKIIEKKQPVAFWCNPISGHNTLYWYLQNLGQGPELLIRYENEEAVDDSQLPKDRFSAERLKGVDSTLKIQPAELGDSAVYLCASSLTRGAEAFFGQGTRLTVVE (SEQ ID NO: 274) MSNQVLCCVVLCFLGANTVDGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSQIVNDFQKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASSRDREQESPLHFGNGTRLTVTE (SEQ ID NO: 275) MGPQLLGYVVLCLLGAGPLEAQVTQNPRYLITVTGKKLTVTCSQNMNHEYMSWYRQDPGLGLRQIYYSMNVEVTDKGDVPEGYKVSRKEKRNFPLILESPSPNQTSLYFCASSFSGGTYEQYFGPGTRLTVTE (SEQ ID NO: 276) MLSPDLPDSAWNTRLLCHVMLCLLGAVSVAAGVIQSPRHLIKEKRETATLKCYPIPRHDTVYWYQQGPGQDPQFLISFYEKMQSDKGSIPDRFSAQQFSDYHSELNMSSLELGDSALYFCASSYRGGSTYEQYFGPGTRLTVTE (SEQ ID NO: 277) MSTRLLCWMALCLLGAELSEAEVAQSPRYKITEKSQAVAFWCDPISGHATLYWYRQILGQGPELLVQFQDESVVDDSQLPKDRFSAERLKGVDSTLKIQPAELGDSAMYLCASSQRDSPNEKLFFGSGTQLSVLE (SEQ ID NO: 278) MGCRLLCCAVLCLLGAVPMETGVTQTPRHLVMGMTNKKSLKCEQHLGHNAMYWYKQSAKKPLELMFVYSLEERVENNSVPSRFSPECPNSSHLFLHLHTLQPEDSALYLCASSQDPYKLSGNTIYFGEGSWLTVVE (SEQ ID NO: 279) DLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 280) DLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG (SEQ ID NO: 281) MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVNIGNHDMRFGAGTRLTVKPN (SEQ ID NO: 282) MEKMLECAFIVLWLQLGWLSGEDQVTQSPEALRLQEGESSSLNCSYTVSGLRGLFWYRQDPGKGPEFLFTLYSAGEEKEKERLKATLTKKESFLHITAPKPEDSATYLCAVQTMDGNQFYFGTGTSLTVIPN (SEQ ID NO: 283) MACPGFLWALVISTCLEFSMAQTVTQSQPEMSVQEAETVTLSCTYDTSESDYYLFWYKQPPSRQMILVIRQEAYKQQNATENRFSVNFQKAAKSFSLKISDSQLGDAAMYFCASSPGTYKYIFGTGTRLKVLAN (SEQ ID NO: 284) MTRVSLLWAVVVSTCLESGMAQTVTQSQPEMSVQEAETVTLSCTYDTSESNYYLFWYKQPPSRQMILVIRQEAYKQQNATENRFSVNFQKAAKSFSLKISDSQLGDTAMYFCAFNPWENYGQNFVFGPGTRLSVLPY (SEQ ID NO: 285) IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 286) MGCRLLCCAVLCLLGAVPIDTEVTQTPKHLVMGMTNKKSLKCEQHMGHRAMYWYKQKAKKPPELMFVYSYEKLSINESVPSRFSPECPNSSLLNLHLHALQPEDSALYLCASSQGTSGADTQYFGPGTRLTVLE (SEQ ID NO: 287) MSIGLLCCAALSLLWAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYSLWDLQETQYFGPGTRLLVLE (SEQ ID NO: 288) MGTSLLCWMALCLLGADHADTGVSQDPRHKITKRGQNVTFRCDPISEHNRLYWYRQTLGQGPEFLTYFQNEAQLEKSRLLSDRFSAERPKGSFSTLEIQRTEQGDSAMYLCASSFSDGGATDTQYFGPGTRLTVLE (SEQ ID NO: 289) MLLLLLLLGPAGSGLGAVVSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLMATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSARPHSLTDTQYFGPGTRLTVLE (SEQ ID NO: 290) DLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG (SEQ ID NO: 291)

[0283] In some embodiments, the TCR construct comprises a human papillomavirus (HPV)-specific TCR chain. In some embodiments, the TCR construct comprising an HPV-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the HPV18 E6 protein and / or the HPV18 E7 protein. In some embodiments, the HPV 18 E6 epitope is amino acids 121-135 and / or amino acids 77-91 of the HPV 18 E6 protein. In some embodiments, the TCR construct comprising an HPV-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the HPV 18 E7 protein. In some embodiments, the HPV 18 E7 epitope is amino acids 11-19. In some embodiments, the HPV-specific TCR sequence, TCR variable domain sequence, CDR sequence, and / or TCR constant domain sequence are described in International Publication No. WO2015 / 009604A1, which is incorporated herein by reference for the purposes described herein.

[0284] In some embodiments, the TCR construct comprises a Kirsten rat sarcoma virus (KRAS)-specific TCR chain. In some embodiments, a TCR construct comprising a KRAS-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the epitope GADGVGKSA (SEQ ID NO: 293). In some embodiments, a TCR construct comprising a KRAS-specific TCR chain comprises a TCR alpha chain and a TCR beta chain that target the epitope GADGVGKSAL (SEQ ID NO: 292). In some embodiments, the KRAS-specific TCR sequence, TCR variable domain sequence, CDR sequence, and / or TCR constant domain sequence are described in U.S. Patent No. 10,611,816B2, which is incorporated herein by reference for the purposes described herein. In some embodiments, the leader sequence and / or signal peptide may be removed from the TCR amino acid sequence, and the percent sequence identity may be calculated based on the TCR amino acid sequence without the leader sequence and / or signal peptide. In some embodiments, the KRAS-specific TCR targets KRAS antigens associated with cancer states including, but not limited to, G12C, G12D, and / or G12R.

[0285] In some embodiments, a TCR construct comprising a KRAS - specific TCR chain 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: 294 - 298. In some embodiments, a TCR construct comprising a KRAS - specific TCR chain 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: 299 - 317. In some embodiments, a TCR construct comprising a KRAS - specific TCR chain 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: 299 - 303. atgggcaccaggctcttcttctatgtggccctttgtctgctgtgggcaggacacagggatgctggaatcacccagagcccaagatacaagatcacagagacaggaaggcaggtgaccttgatgtgtcaccagacttggagccacagctatatgttctggtatcgacaagacctgggacatgggctgaggctgatctattactcagcagctgctgatattacagataaaggagaagtccccgatggctatgttgtctccagatccaagacagagaatttccccctcactctggagtcagctacccgctcccagacatctgtgtatttctgcgccagcagtgaccccggcactgaagctttctttggacaaggcaccagactcacagttgta(SEQ ID NO: 294) atgggcaccaggctcttcttctatgtggccctttgtctgctgtgggcaggacacagggatgctggaatcacccagagcccaagatacaagatcacagagacaggaaggcaggtgaccttgatgtgtcaccagacttggagccacagctatatgttctggtatcgacaagacctgggacatgggctgaggctgatctattactcagcagctgctgatattacagataaaggagaagtccccgatggctatgttgtctccagatccaagacagagaatttccccctcactctggagtcagctacccgctcccagacatctgtgtatttctgcgccagcagtgaccccggcactgaagctttctttggacaaggcaccagactcacagttgtaGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC(SEQ ID NO: 295) atgaaatccttgagagttttactagtgatcctgtggcttcagttgagctgggtttggagccaacagaaggaggtggagcagaattctggacccctcagtgttccagagggagccattgcctctctcaactgcacttacagtgaccgaggttcccagtccttcttctggtacagacaatattctgggaaaagccctgagttgataatgttcatatactccaatggtgacaaagaagatggaaggtttacagcacagctcaataaagccagccagtatgtttctctgctcatcagagactcccagcccagtgattcagccacctacctctgtgccgcggcgatggatagcagctataaattgatcttcgggagtgggaccagactgctggtcaggcct(SEQ ID NO: 296) atgaaatccttgagagttttactagtgatcctgtggcttcagttgagctgggtttggagccaacagaaggaggtggagcagaattctggacccctcagtgttccagagggagccattgcctctctcaactgcacttacagtgaccgaggttcccagtccttcttctggtacagacaatattctgggaaaagccctgagttgataatgttcatatactccaatggtgacaaagaagatggaaggtttacagcacagctcaataaagccagccagtatgtttctctgctcatcagagactcccagcccagtgattcagccacctacctctgtgccgcggcgatggatagcagctataaattgatcttcgggagtgggaccagactgctggtcaggcctATCCAGAACCCCGACCCCGCCGTGTACCAGCTGCGGGACAGCAAGAGCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGATAAGTGCGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAACAGCGCCGTGGCCTGGTCCAACAAGAGCGACTTCGCCTGCGCCAACGCCTTCAACAACAGCATCATCCCCGAGGACACATTCTTCCCAAGCCCCGAGAGCAGCTGCGACGTGAAGCTGGTGGAGAAGTCCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGTCCGTGATCGGCTTCAGAATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGCGGCTGTGGTCCAGC(SEQ ID NO: 297) MGTRLFFYVALCLLWAGHRDAGITQSPRYKITETGRQVTLMCHQTWSHSYMFWYRQDLGHGLRLIYYSAAADITDKGEVPDGYVVSRSKTENFPLTLESATRSQTSVYFCASSDPGTEAFFGQGTRLTVV (SEQ ID NO: 299) MGTRLFFYVALCLLWAGHRDAGITQSPRYKITETGRQVTLMCHQTWSHSYMFWYRQDLGHGLRLIYYSAAADITDKGEVPDGYVVSRSKTENFPLTLESATRSQTSVYFCASSDPGTEAFFGQGTRLTVVEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG (SEQ ID NO: 300) MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAAAMDSSYKLIFGSGTRLLVRP (SEQ ID NO: 301) MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAAAMDSSYKLIFGSGTRLLVRPIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 302) MGTRLFFYVALCLLWAGHRDAGITQSPRYKITETGRQVTLMCHQTWSHSYMFWYRQDLGHGLRLIYYSAAADITDKGEVPDGYVVSRSKTENFPLTLESATRSQTSVYFCASSDPGTEAFFGQGTRLTVVEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRGATNFSLLKQAGDVEENPGPMKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAAAMDSSYKLIFGSGTRLLVRPIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS(SEQ ID NO: 303) MRQVARVIVFLTLSTLSLAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGDMDQAGTALIFGKGTTLSVSS(SEQ ID NO: 304) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLGEGRVDGYTFGSGTRLTVV(SEQ ID NO: 305) MRQVARVIVFLTLSTLSLAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGDMDQAGTALIFGKGTTLSVSS (SEQ ID NO: 306) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLGRASNQPQHFGDGTRLSIL (SEQ ID NO: 307) MRQVARVIVFLTLSTLSLAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGDRDQAGTALIFGKGTTLSVSS (SEQ ID NO: 308) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSFGQSSTYGYTFGSGTRLTVV (SEQ ID NO: 309) MRQVARVIVFLTLSTLSLAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGDMDQAGTALIFGKGTTLSVSSDIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 310) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLGEGRVDGYTFGSGTRLTVVEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVCTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVMAMVKRKNS (SEQ ID NO: 311) MRQVARVIVFLTLSTLSLAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGDMDQAGTALIFGKGTTLSVSSDIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 312) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLGRASNQPQHFGDGTRLSILEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVCTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVMAMVKRKNS (SEQ ID NO: 313) MRQVARVIVFLTLSTLSLAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGDRDQAGTALIFGKGTTLSVSSDIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 314) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSFGQSSTYGYTFGSGTRLTVVEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVCTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVMAMVKRKNS (SEQ ID NO: 315) MKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAAAMDSSYKLIFGSGTRLLVRPDIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 316) MGTRLFFYVALCLLWAGHRDAGITQSPRYKITETGRQVTLMCHQTWSHSYMFWYRQDLGHGLRLIYYSAAADITDKGEVPDGYVVSRSKTENFPLTLESATRSQTSVYFCASSDPGTEAFFGQGTRLTVVEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVCTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVMAMVKRKNS (SEQ ID NO: 317)

[0286] NK cells modified to express the TCR / CD3 receptor complex can be obtained from any suitable source, including fresh or frozen. In certain embodiments, the NK cells are not NK cells obtained from iPSC differentiation. In certain embodiments, the NK cells are obtained by methods well known in the art from human peripheral blood mononuclear cells (PBMCs), non-stimulated leukocyte products (PBSCs), NK cell lines (e.g., NK-92), human embryonic stem cells (hESCs), induced pluripotent stem cells (iPSCs), bone marrow, or umbilical cord blood. Specifically, the NK cells can be isolated from umbilical cord blood (CB), peripheral blood (PB), bone marrow, stem cells, NK cell lines, or mixtures thereof. In certain embodiments, the NK cells are isolated from pooled CB. The CB can be pooled from 2, 3, 4, 5, 6, 7, 8, 9, 10, or more units. The NK cells can be autologous or allogeneic with respect to the recipient individual. The isolated NK cells can or cannot be haplotype-matched to the subject to be treated with the cell therapy. The NK cells can be detected, for example, in humans by specific surface markers such as CD16 and CD56. In some cases, the source of the NK cells is umbilical cord blood, and the NK cells are present as a heterogeneous mixture of cells in the umbilical cord blood, and certain cells expressing CD3 may be depleted. In other methods, umbilical cord CB is used to induce NK cells by isolation of CD34+ cells.

[0287] NK cells can be pre-activated with one or more inflammatory cytokines and can or cannot be expanded. In some cases, NK cells are pre-activated either before being modified to express CD3±TCR or after being modified to express the CD3±TCR complex. In a specific embodiment, pre-activating NK cells can include culturing isolated NK cells in the presence of one or more cytokines. NK cells can be stimulated with IL-2 or other cytokines that bind to the common gamma chain (such as IL-7, IL-12, IL-15, IL-18, IL-21 and others). In a particular embodiment, the cytokine for pre-activation can be selected from the group consisting of IL-12, IL-15, IL-18, and combinations thereof. One or more additional cytokines can be used in the pre-activation step. Pre-activation can be for a short time of 5 to 72 hours, such as 10 to 50 hours, particularly 10 to 20 hours, such as 12, 13, 14, 15, 16, 17, 18, 19 or 20 hours, specifically about 16 hours. The pre-activation culture can contain IL-12 at a concentration of 0.1 to 150 ng / mL, such as 0.5 to 50 ng / mL, particularly 1 to 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. The pre-activation culture can contain IL-18 and / or IL-15 at a concentration of 10 to 100 ng / mL, such as 40 to 60 ng / mL, particularly 45, 46, 47, 48, 49, 50, 51, 52, 53, 54 or 55 ng / mL, specifically about 50 ng / mL.

[0288] In some cases, NK cells are expanded before being modified to express the CD3±TCR complex or after being modified to express the CD3±TCR complex. Pre-activated NK cells can be expanded in the presence of artificial antigen-presenting cells (aAPCs) and / or feeder / fragments or NK activation beads. Pre-activated NK cells can be washed before expansion, such as 2, 3, 4, or 5 times, specifically 3 times. aAPCs can be engineered to express CD137 ligand and / or membrane-bound cytokines. The membrane-bound cytokine can be membrane-bound IL-21 (mIL-21) or membrane-bound IL-15 (mIL-15). In certain embodiments, aAPCs are engineered to express CD137 ligand and mIL-21. aAPCs can be derived from cancer cells such as leukemia cells. aAPCs may not express endogenous HLA class I, II, or CD1d molecules. They can express ICAM-1 (CD54) and LFA-3 (CD58). In particular, aAPCs can be K562 cells such as K562 cells engineered to express CD137 ligand and mIL-21. aAPCs can be irradiated. In some embodiments, fragments of APCs can be used to expand NK cells. The engineering can be performed by any method known in the art, such as retroviral transduction. Retroviral transduction can be performed 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 after co-culturing NK with antigen-presenting cells. In some embodiments, retroviral transduction involves co-transduction of more than one construct. In some embodiments, retroviral transduction occurs after co-culturing with antigen-presenting cells or on about day 5. In some embodiments, co-culturing with antigen-presenting cells continues after transduction of NK cells. Expansion can be for about 2 to 30 days, such as 3 to 20 days, particularly 12 to 16 days, for example 12, 13, 14, 15, 16, 17, 18, or 19 days, specifically about 14 days. Pre-activated NK cells and aAPCs can be present in a ratio of about 3:1 to 1:3, such as 2:1, 1:1, 1:2, specifically about 1:2.The expansion culture may further contain cytokines that promote expansion, such as IL-2. 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. IL-2 can be replenished into the expansion culture, for example, every 2 - 3 days. aAPC can be added to the culture, at least at the second time, for example, on about the 7th day of expansion.

[0289] In certain 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, which are secreted by the NK cells or immobilized on the NK cell membrane. In such cases, the membrane-bound cytokines can be immobilized on the NK cell membrane with specific transmembrane domains, such as the transmembrane domains of CD8, CD28, CD27, B7H3, IgG1, IgG4, CD4, DAP10, DAP12.

[0290] After preparation, the modified NK cells can be immediately injected (e.g., together with an effective amount of one or more monospecific antibodies, bispecific antibodies, or multispecific antibodies), or the NK cells can be stored, such as by cryopreservation. In some cases, when the NK cells are supplied from cryopreservation, the NK cells are inactivated with an inactivating agent (e.g., a kinase inhibitor, such as dasatinib, nilotinib, rapamycin, etc.) before cryopreservation. In certain embodiments, the cells can be expanded ex vivo as a bulk population for several days, weeks, or months within about 1, 2, 3, 4, or 5 days.

[0291] C. Loading of NK cells In certain embodiments, the NK cells are loaded with an antibody prior to use. The NK cells can be loaded in any particular method including immediately prior to culturing or injection, for example, to generate a complex of the NK cells and the antibody. The conditions are sufficient for an effective amount of the antibody to bind to the surface of the NK cells. When using a monospecific antibody, the Fc region of the monospecific antibody binds to the NK cells, while the antigen-binding domain of the monospecific antibody binds freely to the target antigen. In certain embodiments when using a multispecific antibody, one or more antigen-binding domains of the antibody can bind to the surface of the NK cells, such as via an antigen on the surface of the NK cells (e.g., CD3, NKp30, NKp44, NKp46, CD16, CD32, CD64, KIR, etc., but not limited thereto), and the other antigen-binding domains can bind freely to its target antigen. In certain embodiments when using a multispecific antibody, one or more antigen-binding domains of the antibody can bind to one or more target antigens.

[0292] The culture conditions under which NK cells are loaded may or may not be of a specific type having one or more specific parameters. In certain embodiments, the loading of NK cells occurs during culture at a specific temperature such as 37°C, but in alternative embodiments, the temperature is 36°C or 38°C, or lower or higher. The duration of the loading step may be any suitable time, such as in the range from 1 minute to 24 hours or more. For example, it may be in the range of 1 minute to 24 hours, 1 minute to 18 hours, 1 minute to 12 hours, 1 minute to 6 hours, 1 minute to 1 hour, 30 minutes to 24 hours, 30 minutes to 18 hours, 30 minutes to 12 hours, 30 minutes to 6 hours, 30 minutes to 1 hour, 1 to 24 hours, 1 to 18 hours, 1 to 12 hours, 1 to 6 hours, 6 to 24 hours, 6 to 18 hours, 6 to 12 hours, 12 to 24 hours, 12 to 18 hours, or 18 to 24 hours. In some embodiments, the duration of the loading step can be about 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 more, or any range derivable therefrom. In certain embodiments, the cell culture medium is a basal medium or a complex medium. Optionally, the culture may or may not contain one or more reagents utilized during the pre-activation and / or proliferation steps. In certain embodiments, the culture contains one or more cytokines, including, for example, one or more of IL-12, IL-15, IL-2, and IL-18. In some embodiments, the culture contains APCs of any kind.

[0293] In certain embodiments, the antibodies of the compositions described herein are provided in an effective amount to an effective amount of the NK cells of the present disclosure, thereby producing a "chimeric antigen receptor-like" complex. In particular, the antigen-binding domain of the antibody binds to the NK cells, such as via an antigen that is a cell surface protein. A plurality of antibodies can be provided to a plurality of NK cells such that a plurality of cell / antibody complexes are present. The antibody can be of any type, such as monospecific, bispecific, or multispecific. In certain cases, the antibody is involved with both the NK cells and the target antigen via the antigen-binding domain of the antibody (such as an engager in the art that is a fusion protein consisting of two single-chain variable fragments (scFv) of different antibodies). In an example where the antibody is monospecific, the antigen-binding domain of the antibody binds to a target antigen such as a cancer antigen, and another portion of the antibody (such as the Fc region of the antibody) binds to the NK cells. When the antibody is multispecific, one or more antigen-binding domains of the antibody bind to the NK cells (such as via a naturally occurring or genetically introduced NK cell surface antigen such as CD3), and one or more antigen-binding domains of the antibody bind to one or more target antigens. The multispecific antibody can be, for example, bispecific, trispecific, or tetra-specific. When the antibody is trispecific or tetra-specific, the additional antigen-binding domains can bind to other cells such as stem cells.

[0294] In certain embodiments, the antibody can bind to any NK cell surface antigen (which may or may not be a receptor), such as CD16 (including CD16a or CD16b), CD32, CD56, CD64, NKG2D, NKG2C and other c-type lectins, CS1, DNAM, 2B4, CD2 and other costimulatory molecules, NCR, NKp30, NKp44, NKp46, or KIR on the NK cells, direct the NK cells to the target, and enhance the reactivity and specificity against different tumors. In certain embodiments, the antibody targets TROP-2 (tumor-associated calcium signal transducer 2).

[0295] In some embodiments, the antibody can bind to any suitable antigen (such as those described herein, such as those described as targets for CARs and / or TCRs). In certain embodiments, the antibody targets CD19. In certain embodiments, the antibody targets CD20. In certain embodiments, the antibody targets CD123. In certain embodiments, the antibody targets EGFR. In certain embodiments, the antibody targets EGFR2. In certain embodiments, the antibody targets C-met (tyrosine protein kinase MET, also known as mesenchymal-epithelial transition factor). In certain embodiments, the antibody targets EGFR and C-met. In certain embodiments, the antibody targets TROP-2 (tumor-associated calcium signal transducer 2).

[0296] In certain embodiments, the generation of loaded NK cells can be performed by any suitable means such that the appropriate region of the antibody binds to the appropriate surface region of the NK cell. In one example, a particular medium can also be utilized. In certain cases, Plasma-Lyte A and / or human serum albumin are utilized, although this may not always be the case. When the complex is formed in culture, it may or may not be washed prior to administration to the subject, such as by infusion. In another embodiment, the NK cells and the antibody are administered separately and the complex is formed in vivo.

[0297] D. Pre-activation In certain embodiments, the NK cells are pre-activated before administration to the recipient individual. The pre-activation step may be performed before any expansion step or may not be performed. In a specific embodiment, the NK cells are pre-activated with one or more cytokines, and in a specific embodiment, the NK cells are pre-activated with one or more of IL-12, IL-15, IL-2, and IL-18, including two, three, or more. If not all three of IL-12, IL-15, IL-2, and IL-18 are utilized, it may be 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 used in place of IL-15.

[0298] In certain embodiments, the pre-activated cytokines can be IL-12, IL-15, and IL-18. One or more additional cytokines can be used in the pre-activation step. The pre-activation can be for a short time of 5 to 72 hours, such as 10 to 50 hours, particularly 10 to 20 hours, for example 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, or 20 hours, and specifically can be about 16 hours in some cases. The pre-activated culture can contain IL-18 and / or IL-15 at a concentration of 10 to 100 ng / mL, such as 40 to 60 ng / mL, particularly 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 ng / mL, specifically about 50 ng / mL. In some cases, the pre-activated culture contains IL-12 at a concentration of 0.1 to 150 ng / mL, such as at a concentration of 1 to 20 ng / mL, for example 10 ng / mL. In alternative embodiments, the NK cells can be stimulated with IL-2, or other cytokines that bind to the common gamma chain (e.g., IL-7, IL-21, and others), which can be in addition to, or as an alternative to, one or more of IL-12, IL-15, and IL-18. In such cases, the pre-activated culture can contain IL-12 at a concentration of 0.1 to 150 ng / mL, such as 0.5 to 50 ng / mL, particularly 1 to 20 ng / mL, for example 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ng / mL, particularly about 10 ng / mL.

[0299] E. Proliferation In certain 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 prior to the expansion step. The NK cell expansion step may be any suitable one in which the NK cell population is expanded, but in certain embodiments, the expansion step utilizes one or more specific reagents, such as in a culture, to enhance the expansion. In certain embodiments, the NK cells may not be expanded. IL-2 or IL-15 or IL-18 or any combination of these cytokines may be added to the expansion culture before or during expansion. In a specific embodiment, the NK cells can be expanded ex vivo in one of several different bioreactor configurations involving continuous perfusion of the flask or media / additives.

[0300] In certain embodiments, NK cells (regardless of whether they are pre-activated or not) may be washed one, two, three, four, or five times, particularly three times, etc., before and / or after proliferation (e.g., with PBS or Plasma Lyte or human serum albumin or culture medium or combinations thereof). In some embodiments, the cells are washed particularly three times. In certain embodiments, the NK cells are expanded in the presence of artificial antigen-presenting cells (aAPCs). In certain embodiments, the NK cells are expanded in the presence of fragments of aAPCs. The aAPCs can be engineered to express CD137 ligand and / or membrane-bound cytokines. The membrane-bound cytokine may be membrane-bound IL-21 (mIL-21) or membrane-bound IL-15 (mIL-15). In certain embodiments, the aAPC is engineered to express CD137 ligand and mIL-21. The aAPC may be derived from cancer cells such as leukemia cells. The aAPCs may not express endogenous HLA class I, II, or CD1d molecules. They may express ICAM-1 (CD54) and LFA-3 (CD58) or CD48. In particular, the aAPC may be a K562 cell, such as a K562 cell engineered to express CD137 ligand and mIL-21. Methods known in the art, such as retroviral transduction, can be used for the engineering, but any viral or non-viral vector can be utilized. The aAPCs may or may not be irradiated. The expansion may be for a specific period, such as about 2 to 30 days, such as 3 to 20 days, particularly 12 to 16 days, such as 12, 13, 14, 15, 16, 17, 18, or 19 days, particularly about 14 days. The pre-activated NK cells and aAPCs may be present in a ratio of about 3:1 to 1:3, such as 2:1, 1:1, 1:2, particularly about 1:2. The expansion culture may further contain one or more cytokines, such as IL-2, to promote expansion. IL-2 may be present at a concentration of about 10 to 500 U / mL, such as 100 to 300 U / mL, particularly about 200 U / mL. IL-2 can be replenished into the expansion culture at regular intervals, such as every 2 to 3 days. The aAPCs can be added to the culture at least the second time, such as on about day 7.The cytokine used in the pre-activation and / or proliferation process may be a recombinant human cytokine.

[0301] In some embodiments, after proliferation, the NK cells may be utilized immediately in any manner, such as by complexing with one or more antibodies, or may be stored by methods such as cryopreservation. In one aspect, the cells can be expanded ex vivo as a bulk population for several days, weeks, or months within about 1, 2, 3, 4, or 5 days.

[0302] Activated and / or proliferated NK cells secrete type I cytokines such as interferon-γ, tumor necrosis factor-α, granulocyte macrophage colony-stimulating factor (GM-CSF), activate both innate and adaptive immune cells, and also activate other cytokines and chemokines. By measuring these cytokines, the activation state of NK cells can be known. In addition, other methods known in the art for determining NK cell activation can be used for the characterization of the NK cells of the present disclosure.

[0303] Accordingly, with respect to certain pre-activation and proliferation aspects of the present disclosure, in certain embodiments, NK cells pre-activated with any combination of IL-12, IL15, and / or IL-18 and then expanded with aAPC such as K562 cells expressing mIL-21 and CD137 ligand provide very potent cell products. Accordingly, methods of using the NK cells of the present invention for the treatment of various diseases, such as immunotherapy of cancer patients, are provided. In an exemplary method, isolated NK cells are subjected to pre-activation for a short time, such as about 16 hours, with a combination of cytokines such as interleukin-12 (IL-12), IL-15, and / or IL-18, and then can be subjected to expansion using artificial antigen-presenting cells (aAPC) such as K562 feeder cells expressing membrane-bound IL-21 and CD137 ligand, and / or exogenous IL-2. IL-2, IL-15, IL-18, or a combination of these cytokines can be added to the expansion culture at least a second time.

[0304] F. Cryopreservation In certain cases, the NK cells and / or antibodies of the present disclosure are stored in a cryopreservation medium composition containing at least one cryoprotectant, serum (human serum or animal serum) or a non-serum substitute (not human serum or animal serum), and at least one cytokine and / or at least one growth factor. Optionally, the cryoprotectant is dimethyl sulfoxide (DMSO), glycerin, glycerol, hydroxyethyl starch, or a combination thereof. The non-serum substitute can be any type containing at least thrombocytolysate and / or blood product lysate (e.g., human serum albumin). In embodiments of the composition where one or more (including two or more) cytokines are utilized, the cytokine can be a natural or recombinant or synthetic protein. At least one of the cytokines can be a 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. In embodiments of the serum, the serum can be human serum (including human AB serum) or animal-derived serum such as bovine serum. DMSO and other cryoprotectants, when utilized, can be included at 4-10%, 4-6%, 4-8%, 5-10%, 5-8%, 6-10%, 6-8%, 8-10%, etc. of the composition.For embodiments in which serum is employed, the serum may comprise a composition of 5 to 99%, 5 to 95%, 5 to 90%, 5 to 85%, 5 to 80%, 5 to 75%, 5 to 70%, 5 to 65%, 5 to 60%, 5 to 55%, 5 to 50%, 5 to 45%, 5 to 40%, 5 to 35%, 5 to 30%, 5 to 25%, 5 to 20%, 5 to 15%, 5 to 10%, 10 to 99%, 10 to 95%, 10 to 90%, 10 to 85%, 10 to 80%, 10 to 75%, 10 to 70%, 10 to 65%, 10 to 60%, 10 to 55%, 10 to 50%, 10 to 45%, 10 to 40%, 10 to 35%, 10 to 30%, 10 to 25%, 10 to 20%, 10 to 15%, 20 to 99%, 20 to 95%, 20 to 90%, 20 to 85%, 20 to 80%, 20 to 75%, 20 to 70%, 20 to 65%, 20 to 60%, 20 to 55%, 20 to 50%, 20 to 45%, 20 to 40%, 20 to 35%, 20 to 30%, 20 to 25%, 30 to 99%, 30 to 95%, 30 to 90%, 30 to 85%, 30 to 80%, 30 to 75%, 30 to 70%, 30 to 65%, 30 to 60%, 30 to 55%, 30 to 50%, 30 to 45%, 30 to 40%, 30 to 35%, 40 to 99%, 40 to 95%, 40 to 90%, 40 to 85%, 40 to 80%, 40 to 75%, 40 to 70%, 40 to 65%, 40 to 60%, 40 to 55%, 40 to 50%, 40 to 45%, 50 to 99%, 50 to 95%, 50 to 90%, 50 to 85%, 50 to 80%, 50 to 75%, 50 to 70%, 50 to 65%, 50 to 60%, 50 to 55%, 60 to 99%, 60 to 95%, 60 to 90%, 60 to 85%, 60 to 80%, 60 to 75%, 60 to 70%, 60 to 65%, 70 to 99%, 70 to 95%, 70 to 90%, 70 to 85%, 70 to 80%, 70 to 75%, 80 to 99%, 80 to 95%, 80 to 90%, 80 to 85%, 90 to 99%, 90 to 95%, or 95 to 99%. The composition may also contain at least 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%, or less of the serum.In certain embodiments, the composition may contain thromboplastin at any concentration in the composition. However, in certain embodiments, the thromboplastin is 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 contain at least 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%, or less of thromboplastin.

[0305] The composition can have components at specific concentrations including cytokines and / or growth factors. In specific cases, for example, any cytokine including IL-2, IL-21, and / or IL-15 is present in the composition at a specific concentration. IL-2 can be present, for example, at a concentration of 1 to 5000, 1 to 1000, 1 to 500, 1 to 100, 100 to 5000, 100 to 500, 500 to 5000, 500 to 1000, or 1000 to 5000 U / mL. In a specific case, IL-2 is present in the composition at a concentration of at least 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 U / mL, or at a concentration below these. In a specific embodiment, IL-21 is present in the composition at a concentration of 10 to 3000, 10 to 2000, 10 to 1000, 10 to 500, 10 to 100, 100 to 3000, 100 to 2000, 100 to 1000, 500 to 3000, 500 to 2000, 500 to 1000, 1000 to 3000, 1000 to 2000, or 2000 to 3000 ng / mL. IL-21 can be present in the composition at a concentration of at least 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, or at a concentration below these. IL-15 can be present in the composition at a concentration of 1 to 2000, 1 to 1000, 1 to 500, 1 to 100, 100 to 2000, 100 to 1000, 100 to 500, 500 to 2000, 500 to 1000, or 1000 to 2000 ng / mL. IL-15 can be present in the composition at a concentration of at least 10, 50, 100, 500, 1000, 1500, or 2000 ng / mL, or at a concentration below these.

[0306] Compositions as encompassed herein that include at least one cryoprotectant, serum or serum replacement, and at least one cytokine and / or at least one growth factor may further include any of a plurality of types of immune cells and / or stem cells, respectively. 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) that may be derived from bone marrow, peripheral blood, skin, adipose tissue, or combinations thereof, hematopoietic stem cells, induced pluripotent stem cells, MSCs (a cell population also referred to in the literature as "mesenchymal stem cells" and "mesenchymal stromal cells"), or mixtures thereof. In embodiments where NK cells are utilized, the NK cells may or may not be proliferating NK cells. Embodiments of the present disclosure also encompass pharmaceutical compositions that include any of the compositions of the present disclosure and a suitable pharmaceutically acceptable carrier.

[0307] In certain embodiments, the cells and / or antibodies are treated with one or more inactivating agents (e.g., kinase inhibitors such as dasatinib, nilotinib, rapamycin, etc.) prior to cryopreservation.

[0308] In some embodiments, the techniques described herein include inactivating NK cells, which includes treating NK cells with an effective amount of one or more inactivating agents under conditions that produce inactivated NK cells. In some embodiments, the inactivating agent is a kinase inhibitor. In some embodiments, the inactivating 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 inactivating 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, pegdastinib, dacomitinib, osimertinib, erlotinib, gefitinib, lapatinib, afatinib, pemigatinib, erdafitinib, nintedanib, gilteritinib, midostaurin, tucatinib, neratinib, baricitinib, luxolutinib, fedratinib, tofacitinib, repotrectinib, selumetinib, binimetinib, cobimetinib, trametinib, upadacitinib, abemaciclib, serpelitinib, 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.

[0309] In some embodiments, the treatment with the inactivating agent is at any point during the culture of NK cells. In some embodiments, the treatment is from about 24 hours to about 96 hours, from about 36 hours to about 84 hours, or from about 48 hours to about 72 hours. In some embodiments, the treatment is about 24 hours, about 48 hours, or about 72 hours. In some embodiments, the NK cells are treated with an inactivating agent at a concentration of about 1 to about 1000 nM. In some embodiments, the NK cells are treated with an inactivating agent at a concentration of about 5 to about 500 nM. In some embodiments, the NK cells are treated with an inactivating agent at a concentration of about 20 to about 200 nM. In some embodiments, the NK cells are treated with an inactivating agent at a concentration of about 30 to about 100 nM. In some embodiments, the inactivated NK cells have an increase in the expression of one or more of C-kit, CCR-5, CD62L, and / or CXCR4, and / or a decrease in the expression of one or more of NKG2D, DNAM, OX-40, TRAIL, HLA-DR, CD2, CD25, ICOS, and / or CD95 as compared to activated NK cells.

[0310] In some embodiments, the techniques described herein include a method of maintaining the viability of a population of cells at at least 50% or more after cryopreservation of the population, the method comprising subjecting the population to an effective amount of one or more inactivating agents (e.g., tyrosine kinase inhibitors) to inactivate the cells prior to cryopreservation, cryopreserving the cells, and thawing the population, wherein the viability of the population is at least 50% or more upon thawing. Optionally, upon thawing of the cells, the viability of the population of cells is at least 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% or more after cryopreservation of the population.

[0311] III. Heterologous Proteins and Mutations In certain embodiments, the NK cells are modified to express not only one or more components of the TCR / CD3 complex, but also one or more other heterologous proteins. The heterologous proteins can enhance the activity of the NK cells in any manner, including at least their activation, persistence, expansion, homing, and / or cytotoxicity.

[0312] A. Monospecific, bispecific or multispecific antibodies In some embodiments, the NK cells are modified to express one or more monospecific, bispecific or multispecific antibodies, while in other cases the NK cells do not express antibodies but the antibodies are utilized with the NK cells.

[0313] When the NK cells are modified to express antibodies, the antibodies can be engagers that crosslink specific immune effector cells to specific target cells for the destruction of the target cells. The present disclosure enables the use of modified NK cells together with a standard T cell engager (BiTE), which is often modified to express CD3, a T cell antigen to which the BiTE engager binds. In such cases, the BiTE used in the present invention can also target a cancer antigen or a viral antigen that can be adapted to the medical condition of the intended recipient individual. For example, the BiTE can be adapted to bind to a cancer antigen characteristic of the cancer cells of an individual's cancer. The anti-CD3 antibody of the BiTE can target the CD3γ chain, the CD3δ chain, the CD3ε chain, or the CD3ζ chain.

[0314] In some cases, in addition to expressing a CD3 complex (with or without a TCR) that enables the use of NK cells as a therapeutic method with BiTE, NK cells can be modified to express (or not express but instead co-administer) one or more bispecific NK engagers (BiKE). BiKE includes an antibody that binds to a surface protein on NK cells, including surface proteins that are naturally expressed on NK cells (e.g., but not limited to, NKp30, NKp44, NKp46, CD16, CD32, CD64, KIR, etc.), and further includes an antibody that binds to a desired target antigen. BiKE can target NK cells via an antibody to an NK surface protein such as, for example, CD16, CS1, CD32, CD64, CD56, NKG2D, NKG2C, DNAM, 2B4, CD2, NCR, NKp30, NKp44, NKp46, or KIR. In such cases, the BiKE used in the present invention can also target a cancer antigen or a viral antigen that can be adapted according to the medical condition of the intended recipient individual. For example, BiKE can be adapted to bind to a cancer antigen that is characteristic of the cancer cells of an individual's cancer.

[0315] In certain embodiments, the antibody is blinatumomab. In certain embodiments, the antibody is tebentafusp. In certain embodiments, the antibody is mosunetuzumab. In certain embodiments, the antibody is teclistamab. In certain embodiments, the antibody is glofitamab. In certain embodiments, the antibody is epcoritamab. In some embodiments, the antibody is flotetuzumab. In some embodiments, the antibody is APV0436. In some embodiments, the antibody is TNB383B. In certain embodiments when using a multispecific antibody, one or more antigen-binding domains of the antibody can bind to one or more target antigens. In certain embodiments, the antibody is amivantamab. In certain embodiments, the antibody is cetuximab. In certain embodiments, the antibody is imugamozumab.

[0316] In embodiments where NK cells express a CD3 complex (with or without a TCR) and one or more BiKEs, one or more vectors can be utilized to transfect or transduce cells with the CD3 complex components (with or without a TCR) and one or more BiKEs. Optionally, the one or more CD3 complex components (with or without a TCR) and the BiKE can or cannot be on the same multicistronic vector.

[0317] B. Engineered receptors In certain embodiments, NK cells are engineered to express one or more engineered receptors. Optionally, the engineered receptor is an engineered antigen receptor that targets any type of cancer antigen or viral antigen. The receptor can be adapted to target a desired antigen based on the medical condition of the intended recipient individual.

[0318] In some embodiments, the engineered antigen receptor is a chimeric antigen receptor (CAR). NK cells can be modified to encode at least one CAR, and the CAR can be, for example, a first-generation, second-generation, third-generation, or later-generation CAR. The CAR can or cannot be bispecific for two or more different antigens. The CAR can include one or more co-stimulatory domains. Each co-stimulatory domain can include, for example, any one or more of the co-stimulatory domains of a member of the TNF receptor superfamily, CD28, CD137 (4-1BB), CD134 (OX40), DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1 (CD11a / CD18), Lck, TNF-RI, TNF-RII, Fas, CD30, CD27, NKG2D, 2B4M, CD40, or combinations thereof. In a specific embodiment, the CAR includes CD3 zeta. In certain embodiments, the CAR lacks one or more specific co-stimulatory domains; for example, the CAR may lack 4-1BB and / or CD28.

[0319] In certain embodiments, the CAR polypeptide within the cell comprises an extracellular spacer domain that links the antigen-binding domain and the transmembrane domain, which may be referred to as a hinge. The extracellular spacer domain includes, but is not limited to, the Fc fragment of an antibody or a fragment or derivative thereof, the hinge region of an antibody or a fragment or derivative thereof, the CH2 region of an antibody, the CH3 region of an antibody, an artificial spacer sequence, or a combination thereof. Examples of the extracellular spacer domain include, but are not limited to, artificial spacers made from polypeptides such as CD8α hinge, CD28, Gly3, or the CH1, CH3 domains of IgG (such as human IgG1 or IgG4). In certain embodiments, the extracellular spacer domain may include (i) the hinge, CH2, and CH3 regions of IgG4, (ii) the hinge region of IgG4, (iii) the hinge and CH2 of IgG4, (iv) the hinge region of CD8α or CD4, (v) the hinge, CH2, and CH3 regions of IgG1, (vi) the hinge region of IgG1, or (vii) the hinge and CH2 of IgG1, (viii) the hinge region of CD28, or a combination thereof. In certain embodiments, the hinge is derived from IgG1, and in certain embodiments, the CAR polypeptide comprises a specific IgG1 hinge amino acid sequence or is encoded by a specific IgG1 hinge nucleic acid sequence.

[0320] The transmembrane domain of the CAR may be of natural or synthetic origin. In the case of natural origin, in certain embodiments, a domain derived from a membrane-bound protein or transmembrane protein is used. The transmembrane region includes those derived from the alpha, beta, or zeta chains of the T cell receptor, CD28, CD3 zeta, CD3 epsilon, CD3 gamma, CD3 delta, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD30, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154, ICOS / CD278, GITR / CD357, NKG2D, and DAP molecules (such as DAP10 or DAP12) (i.e., those containing at least their transmembrane region). Alternatively, the transmembrane domain in some embodiments is synthetic. In some embodiments, the synthetic transmembrane domain mainly contains hydrophobic residues such as leucine and valine. In some embodiments, a triplet of phenylalanine, tryptophan, and valine may be found at both ends of the synthetic transmembrane domain.

[0321] In some embodiments, the engineered receptor utilizes one or more homing receptors (which can home to a target, such as an event utilizing an adhesion molecule, not necessarily for signal release) 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.

[0322] C. Cytokine In some embodiments, the cells expressing NK cells are engineered to express one or more heterologous cytokines and / or upregulate the normal expression of one or more heterologous cytokines. The cells may or may not be transduced or transfected with one or more cytokines on the same vector as other genes. In certain embodiments, the NK cells can be modified to express one or more cytokines, cytokine receptors, chemokines, chemokine receptors, and / or suicide genes.

[0323] One or more cytokines may be co-expressed from a vector encoding a polypeptide other than the engineered antigen receptor and / or suicide gene. For example, interleukin-15 (IL-15) is tissue-restricted and is only observed in serum or systemically at any level under pathological conditions. IL-15 has several properties desirable for adoptive therapy. IL-15 is a homeostatic cytokine that induces the development and cell proliferation of natural killer cells, promotes the eradication of established tumors by alleviating the functional inhibition of tumor resident cells, and inhibits activation-induced cell death (AICD). In addition to IL-15, other cytokines are envisioned. These include cytokines, chemokines, and other molecules that contribute to the activation and proliferation of cells used for human applications, but are not limited thereto. NK cells expressing IL-15 enable sustained supportive cytokine signaling, which is useful for survival after infusion.

[0324] In some embodiments, the cells express one or more engineered receptors provided exogenously, where the engineered receptors include chemokine receptors and / or cytokine receptors. In some embodiments, the cytokine receptor is the IL-15 receptor. In some embodiments, the cytokine receptor is a non-naturally occurring variant of a cytokine receptor. In some embodiments, the cytokine receptor is the IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, or GMCSF receptor, or a combination thereof.

[0325] In certain embodiments, the cells express one or more exogenously supplied cytokines. By way of example, the cytokines are IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, GMCSF, or combinations thereof. Since the cytokines are expressed from an expression vector within the cell, they may be exogenously supplied to the NK cells. In another case, endogenous cytokines within the cell are upregulated by manipulation of the expression control of the endogenous cytokines, such as gene recombination at the promoter site of the cytokine. When the cytokine is provided to the cell on an expression construct, the cytokine may be encoded from the same vector as the suicide gene and / or the CAR. In some embodiments, the present disclosure relates to the co-utilization of IL-15 with the CAR and optionally a suicide gene.

[0326] In some embodiments, a specific sequence of IL-15 is utilized as follows (the underline means the signal peptide sequence): ATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCC GGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC (SEQ ID NO: 49) MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS (SEQ ID NO: 48)

[0327] The modified NK cells of the present disclosure are utilized with a monospecific, bispecific, or multispecific antibody that targets one or more specific antigens. Additionally, the NK cells can be modified with an engineered antigen receptor that targets one or more specific antigens. When the NK cells are modified with one or more engineered antigen receptors, the antigen targeted by the monospecific, bispecific, or multispecific antibody and the antigen targeted by the one or more engineered antigen receptors can be the same antigen or can be different antigens. In some cases, the antigen targeted by the monospecific, bispecific, or multispecific 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. In some embodiments, the antigen targeted by the monospecific, bispecific, or multispecific antibody and the antigen targeted by the one or more engineered antigen receptors are different antigens but are each associated with solid tumors.

[0328] Among the antigens targeted by the genetically engineered antigen receptor are those expressed in the context of a disease, condition, or cell type targeted via adoptive cell therapy. Among the diseases and conditions are proliferative, neoplastic, and malignant diseases and disorders, including cancers of the immune system such as blood cancers, 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, such as tumor or pathogenic cells, compared to normal or non-target cells or tissues. In other embodiments, the antigen is expressed on normal cells and / or on the engineered cells.

[0329] In some embodiments, the antigen can be presented in the context of an HLA protein. In some embodiments, the antigen can be presented in the context of the HLA-Cw0802 protein. In some embodiments, target cells that express the antigen of interest can express the antigen transgenically, express the proteins required for antigen presentation transgenically, and / or be loaded with the antigen. In some embodiments, the target cells are transfected with an HLA-Cw0802 construct and / or a b2m construct that comprises a polynucleotide 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: 318-323 or encodes an amino acid sequence. In some embodiments, the target cells are loaded with a peptide comprising the epitope GADGVGKSA (SEQ ID NO: 293) and / or GADGVGKSAL (SEQ ID NO: 294). ATGTCTCGCTCCGTGGCCTTAGCTGTGCTCGCGCTACTCTCTCTTTCTGGCCTGGAGGCTATCCAGCGTACTCCAAAGATTCAGGTTTACTCACGTCATCCAGCAGAGAATGGAAAGTCAAATTTCCTGAATTGCTATGTGTCTGGGTTTCATCCATCCGACATTGAAGTTGACTTACTGAAGAATGGAGAGAGAATTGAAAAAGTGGAGCATTCAGACTTGTCTTTCAGCAAGGACTGGTCTTTCTATCTCTTGTACTACACTGAATTCACCCCCACTGAAAAAGATGAGTATGCCTGCCGTGTGAACCATGTGACTTTGTCACAGCCCAAGATAGTTAAGTGGGATCGAGACATG (SEQ ID NO: 319) MRVMAPRTLILLLSGALALTETWACSHSMRYFYTAVSRPGRGEPRFIAVGYVDDTQFVQFDSDAASPRGEPRAPWVEQEGPEYWDRETQKYKRQAQTDRVSLRNLRGYYNQSEAGSHTLQRMYGCDLGPDGRLLRGYNQFAYDGKDYIALNEDLRSWTAADKAAQITQRKWEAAREAEQRRAYLEGTCVEWLRRYLENGKKTLQRAEHPKTHVTHHPVSDHEATLRCWALGFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPEPLTLRWGPSSQPTIPIVGIVAGLAVLAVLAVLGAVMAVVMCRRKSSGGKGGSCSQAASSNSAQGSDESLIACKA (SEQ ID NO: 321) MSRSVALAVLALLSLSGLEAIQRTPKIQVYSRHPAENGKSNFLNCYVSGFHPSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEYACRVNHVTLSQPKIVKWDRDM (SEQ ID NO: 322) MRVMAPRTLILLLSGALALTETWACSHSMRYFYTAVSRPGRGEPRFIAVGYVDDTQFVQFDSDAASPRGEPRAPWVEQEGPEYWDRETQKYKRQAQTDRVSLRNLRGYYNQSEAGSHTLQRMYGCDLGPDGRLLRGYNQFAYDGKDYIALNEDLRSWTAADKAAQITQRKWEAAREAEQRRAYLEGTCVEWLRRYLENGKKTLQRAEHPKTHVTHHPVSDHEATLRCWALGFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPEPLTLRWGPSSQPTIPIVGIVAGLAVLAVLAVLGAVMAVVMCRRKSSGGKGGSCSQAASSNSAQGSDESLIACKAATNFSLLKQAGDVEENPGPMSRSVALAVLALLSLSGLEAIQRTPKIQVYSRHPAENGKSNFLNCYVSGFHPSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEYACRVNHVTLSQPKIVKWDRDM(SEQ ID NO: 323)

[0330] In this method, any suitable antigen can be targeted. The antigen may, in some cases, be related to specific cancer cells but not to non-cancer cells. Exemplary antigens include, but are not limited to, antigen molecules derived from infectious agents, self / self-antigens, tumor / cancer-related antigens, tumor neoantigens (Linnemann et al., 2015). In certain embodiments, the antigen includes KRAS, TROP-2, NY-ESO, CD19, EBNA, CD123, HER2, CA-125, TRAIL / DR4, CD20, CD22, CD70, CD38, CD123, CLL1, carcinoembryonic antigen, alpha-fetoprotein, CD56, AKT, Her3, epithelial tumor antigen, CD319 (CS1), ROR1, folate-binding protein, HIV-1 envelope glycoprotein gp120, HIV-1 envelope glycoprotein gp41, CD5, CD23, CD30, HERV-K, IL-11Rα, kappa chain, lambda chain, CSPG4, CD33, CD47, CLL-1, U5snRNP200, CD200, BAFF-R, BCMA, CD99, p53, mutant p53, Ras, mutant ras, c-Myc, cytoplasmic serine / threonine kinase (e.g., A-Raf, B-Raf, and C-Raf, cyclin-dependent kinase), MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A10, MAGE-A12, MART-1, melanoma-related antigen, BAGE, DAM-6, -10, GAGE-1, -2, -8, GAGE-3, -4, -5, -6, -7B, NA88-A, MC1R, mda-7, gp75, Gp100, PSA, PSM, tyrosinase, tyrosinase-related protein, TRP-1, TRP-2, ART-4, CAMEL, CEA, Cyp-B, hTERT, hTRT, iCE, MUC1, MUC2, phosphoinositide 3 kinase (PI3K), TRK receptor, PRAME, P15, RU1, RU2, SART-1, SART-3, Wilms tumor antigen (WT1), AFP, beta-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 Kinase (e.g., Epidermal Growth Factor Receptor (EGFR) (in certain embodiments, EGFRvIII), Platelet-Derived Growth Factor Receptor (PDGFR), Vascular Endothelial Growth Factor Receptor (VEGFR)), VEGFR2, Cytoplasmic Tyrosine Kinase (e.g., src family, syk-ZAP70 family), Integrin-Binding Kinase (ILK), Signal Transducer and Activator of Transcription STAT3, STATS, and STATE, Hypoxia-Inducible Factor (e.g., HIF-1 and HIF-2), Nuclear Factor-κB (NF-κB), Notch Receptor (e.g., Notch1-4), NY ESO 1, c-Met, Mammalian Target of Rapamycin (mTOR), WNT, Extracellular Signal-Regulated Kinase (ERK), and their regulatory subunits, PMSA, PR-3, MDM2, Mesothelin, Renal Cell Carcinoma-5T4, SM22α, Carbonic Anhydrase I (CAI) and IX (CAIX) (also known as G250), STEAD, TEL / AML1, GD2, Proteinase 3, hTERT, Sarcoma Translocation Breakpoint, EphA2, ML-IAP, EpCAM, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, ALK, Androgen Receptor, Cyclin B1, Polysialic Acid, MYCN, RhoC, GD3, Fucosyl GM1, Mesothelin, PSCA, sLe, PLAC1, GM3, BORIS, Tn, GloboH, NY-BR-1, RGsS, SAGE, SART3, STn, PAX5, OY-TES1, Spermatogenic Protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE1, B7H3, Legumain, TIE2, Page4, MAD-CT-1, FAP, MAD-CT-2, fos-related antigen 1, CBX2, CLDN6, SPANX, TPTE, ACTL8, ANKRD30A, CDKN2A, MAD2L1, CTAG1B, SUNC1, and LRRN1 are included. Examples of antigen sequences are, for example, GenBank, 登録商標In the database: KRAS (accession number NG_000012.12), CD19 (accession number NG_007275.1), EBNA (accession number NG_002392.2), WT1 (accession number NG_009272.1), CD123 (accession number NC_000023.11:1336785-1382689), NY-ESO (accession number: NC_000023.11), EGFRvIII (accession number: NG_007726.3), MUC1 (accession number: NG_029383.1), HER2 (accession number: NG_007503.1), CA-125 (accession number NG_055257.1), WT1 (accession number NG_009272.1), Mage-A3 (accession number NG_013244.1), Mage-A4 (accession number NG_013245.1), Mage-A10 (accession number NC_000023.11), TRAIL / DR4 (accession number NC_000003.12), and / or CEA (accession number NC_000019.10) are known in the art.

[0331] Tumor-associated antigens can be derived from, for example, prostate cancer, breast cancer, colorectal cancer, lung cancer, pancreatic cancer, kidney cancer, mesothelioma, ovarian cancer, liver cancer, brain tumor, bone cancer, gastric cancer, spleen cancer, testicular cancer, cervical cancer, anal cancer, gallbladder cancer, thyroid cancer, or melanoma. Exemplary tumor-associated antigens or tumor cell-derived antigens include MAGE1, 3, and MAGE4 (or other MAGE antigens as disclosed in International Patent Publication No. WO99 / 40188); PRAME; BAGE; RAGE, Lage (also known as NY ESO 1); KRAS (as described in U.S. Patent No. 10611816); 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 cancer, sarcoma, and bladder cancer. See, for example, U.S. Patent No. 6,544,518. Prostate cancer tumor-associated antigens include, for example, prostate-specific membrane antigen (PSMA), prostate-specific antigen (PSA), prostate acid phosphate, NKX3.1, and prostate stem cell antigen 6 (STEAP).

[0332] Other tumor-related antigens include Plu-1, HASH-1, HasH-2, Cripto, Criptin, etc. Furthermore, the tumor antigen may be a self-peptide hormone. For example, gonadotropin-releasing hormone (GnRH), which is a short peptide 10 amino acids in length, is useful for the treatment of many cancers.

[0333] Antigens can include genes mutated in tumor cells, such as telomerase enzyme, survivin, mesothelin, mutant ras, bcr / abl rearrangement, Her2 / neu, mutant or wild-type p53, cytochrome P450 1B1, abnormal expression intron sequences such as N-acetylglucosaminyltransferase-V; clonal rearrangements of immunoglobulin genes that generate unique idiotypes in multiple myeloma and B-cell lymphoma; tumor antigens containing epitopic regions or epitopic peptides derived from tumor virus processes such as human papillomavirus proteins E6 and E7; Epstein-Barr virus protein LMP2; epitopic regions or epitopic peptides derived from genes transcribed at different levels in tumor cells compared to normal cells, such as non-mutated onco-fetal proteins with tumor-selective expression like carcinoembryonic antigen and α-fetoprotein.

[0334] E. Suicide gene In certain embodiments, a suicide gene is utilized with NK cell therapy to control its use and enable the termination of cell therapy at a desired event and / or time. The suicide gene is employed in transduced cells for the purpose of inducing the death of the transduced cells when necessary. Cells of the present disclosure modified to carry one or more vectors encompassed by the present disclosure may contain one or more suicide genes. In some embodiments, the term "suicide gene" as used herein is defined as a gene that, upon administration of a prodrug or other agent, results in the conversion of the gene product into a compound that kills the host cell. In other embodiments, the suicide gene encodes a gene product that is targeted by an agent (such as an antibody) that targets the suicide gene product, if desired.

[0335] In some cases, cell therapy may be a candidate for the use of any type of one or more suicide genes if the individual receiving the cell therapy and / or the individual who has received the cell therapy exhibits one or more symptoms of one or more adverse events such as, by way of example, cytokine release syndrome, neurotoxicity, anaphylaxis / allergic reactions, and / or on-target / off-tumor toxicity, or is at risk of having one or more symptoms, including in cases of urgency. The use of the suicide gene may be part of a planned protocol for treatment or may be used only if the need for its use is recognized. In some cases, the cell therapy may be terminated using an agent that targets the suicide gene or its gene product because the treatment is no longer necessary.

[0336] The use of a suicide gene can be initiated at the point when at least one adverse event occurs to the individual, which adverse event can be recognized by any means, including during regular monitoring which may or may not be continuous since the start of cell therapy. The adverse event can be detected by an examination and / or a test. When the individual develops a cytokine release syndrome (also sometimes called cytokine storm), the individual may present with an elevation of inflammatory cytokine(s) (by way of example only: interferon-γ, granulocyte macrophage colony stimulating factor, IL-10, IL-6 and TNF-α), fever, fatigue, hypotension, hypoxia, tachycardia, nausea, capillary leak, heart / kidney / liver dysfunction, or a combination thereof. When the individual has neurotoxicity, the individual may have confusion, delirium, aphasia, and / or seizures. In some cases, the individual is tested for markers associated with the onset and / or severity of cytokine release syndrome, such as C-reactive protein, IL-6, TNF-α, and / or ferritin.

[0337] Examples of suicide genes include engineered non-secretory (including membrane-bound) tumor necrosis factor (TNF)-α mutant polypeptides (see PCT / US19 / 62009, which is hereby incorporated by reference in its entirety), which may be affected by the delivery of antibodies that bind to TNF-α variants. Examples of suicide gene / prodrug combinations that can 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 deoxycytidine kinase and cytarabine. Escherichia coli purine nucleoside phosphorylase, a so-called suicide gene that converts the prodrug 6-methylpurine deoxyriboside to the toxic purine 6-methylpurine, can be used. Other suicide genes include CD20, CD52, inducible caspase 9, purine nucleoside phosphorylase (PNP), cytochrome p450 enzyme (CYP), carboxypeptidase (CP), carboxylesterase (CE), nitroreductase (NTR), guanine ribosyltransferase (XGRTP), glycosidase enzyme, methionine-α,γ-lyase (MET), thymidine phosphorylase (TP), and the like.

[0338] F. Knockout or knockdown of endogenous genes In certain embodiments, the NK cells of the disclosure can include gene editing of 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 is performed in NK cells that express one or more heterologous transgenes (e.g., CD3, TCR, etc.), while in other cases, the gene editing is performed in NK cells that do not express heterologous transgenes but will ultimately express one or more heterologous transgenes, at least in some cases. In certain embodiments, the NK cells to be gene edited are proliferating NK cells.

[0339] In certain embodiments, one or more endogenous genes of NK cells are modified, such as by disruption of expression where part or all of the expression is decreased. In certain cases, one or more genes are knocked down or knocked out using the processes of the present disclosure. In certain cases, multiple genes are knocked down or knocked out in the same process as the processes of the present disclosure. The genes edited in NK cells can be any genes, but in specific embodiments the genes are genes whose gene products inhibit the activity and / or proliferation of NK cells. In certain cases, the genes edited in NK cells enable the NK cells to function more effectively in the 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, β2-microglobulin, HLA, CD73, CREB, CREM, ICER, and CD39. In specific embodiments, the TGFBR2 gene is knocked out or knocked down in NK cells. In specific embodiments, the CISH gene is knocked out or knocked down in NK cells. In specific embodiments, the CD38 gene is knocked out or knocked down in NK cells.

[0340] In some embodiments, gene editing is performed using one or more DNA-binding nucleic acids, such as modification via an RNA-guided endonuclease (RGEN). For example, the modification can be performed using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins. Generally, the term "CRISPR system" collectively refers to transcripts and other elements involved in directing the expression or activity of CRISPR-associated ("Cas") genes, including sequences encoding Cas genes, tracr (trans-activating CRISPR) sequences (e.g., tracrRNA or the active portion of tracrRNA), tracr-mate sequences (including "direct repeats" and tracrRNA-processing partial direct repeats in the context of endogenous CRISPR systems), guide sequences (also referred to as "spacers" in the context of endogenous CRISPR systems), and / or other sequences and transcripts from the CRISPR locus. Methods of utilizing CRISPR systems are well known in the art.

[0341] IV. Administration of Therapeutic Compositions NK cells expressing CD3 and a monospecific, bispecific or multispecific antibody are administered to an individual in need thereof, including in a manner such that the anti-CD3 antibody of the bispecific or multispecific antibody is in proximity to bind to CD3 on the CD3-expressing NK cells. Optionally, the two components are administered separately to the individual, but in other cases, the two components are complexed prior to administration, such as in an ex vivo manner. In another embodiment, the NK cells express the antibody. Optionally, the two components are not pre-complexed prior to administration, but are co-administered by any suitable route of administration, such as co-injection into the patient.

[0342] Embodiments of the present disclosure relate to methods of using compositions comprising the NK cells and antibodies provided herein for treating or preventing a medical disease or disorder. The method comprises administering to a subject a therapeutically effective amount of CD3(±TCR)-modified NK cells and an antibody, thereby treating or preventing a disease in the subject, including reducing the risk of the disease, reducing the severity of the disease, and / or delaying the onset of the disease. In certain embodiments of the present disclosure, cancer or an infectious disease is treated by transfer of a composition comprising an NK cell population and a corresponding antibody. In at least some cases, the NK cells may enhance the adaptive immune response by reversing the anti-inflammatory tumor microenvironment and promoting the differentiation, activation, and / or recruitment of accessory immune cells to the malignant tumor site due to the release of their inflammatory cytokines. In certain embodiments, the providing step may include culturing the NK cells with the antibody molecule for a specific time (e.g., about 5 minutes to about 24 hours or more), and storing the NK cells and the antibody molecule for a certain period before infusion / administration (e.g., about 1, 2, 3, 4, 5 days, or a period exceeding 5 days).

[0343] The cancers in which the treatment method of the present invention is useful include all malignant cell types such as those found in solid tumors or hematological tumors. Exemplary solid tumors can include, but are not limited to, tumors of organs 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, malignant tumors of T cells or B cells, leukemia, lymphoma, blastoma, myeloma, and the like. Further examples of cancers that can be treated using the methods provided herein include lung cancer (including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and squamous cell lung cancer), peritoneal cancer, gastric cancer or stomach cancer (including gastrointestinal cancer and gastrointestinal stromal cancer), pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, breast cancer, colorectal cancer, endometrial cancer or corpus cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, various head and neck cancers, and melanoma, but are not limited thereto. In certain embodiments, the cancer is ovarian cancer. In certain embodiments, the cancer is lung cancer. In certain embodiments, the cancer is colorectal cancer. In certain embodiments, the cancer is a cancer associated with a KRAS-positive tumor.

[0344] Cancer may be, but is not limited to, the following tissue types: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant cell and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; papilloma of the nipple; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; spongy adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma of adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; malignant carcinoid tumor; bronchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; eosinophilic carcinoma; eosinophilic adenocarcinoma; basophilic carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; nonencapsulating sclerosing carcinoma; adrenocortical carcinoma; endometrial carcinoma; carcinoma of skin appendages; apocrine adenocarcinoma; sebaceous gland carcinoma; keratinizing gland carcinoma; 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, breast; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma with squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; sarcoma, malignant; granulosa cell tumor, malignant; androblastoma, malignant; sertoli cell carcinoma; leydig cell tumor, malignant; lipoid cell tumor, malignant; paraganglioma, malignant; extra-mammary paraganglioma, malignant; pheochromocytoma; angiosarcoma; malignant melanoma; amelanotic melanoma; superficially spreading melanoma; malignant lentigo; acral lentiginous melanoma; nodular melanoma; malignant melanoma of giant pigmented nevus; epithelioid cell melanoma; blue nevus, malignant; sarcoma; fibrosarcoma; malignant fibrous histiocytoma; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mixed tumor, malignant; müllerian mixed tumor; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymal tumor, malignant; Brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; germ cell neoplasia; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; chordoma, malignant; angiosarcoma; angioendothelioma, malignant; kaposi sarcoma; hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma; parosteal osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal chondrosarcoma; giant cell tumor of bone; Ewing sarcoma; odontogenic tumor, malignant; ameloblastic epulis; ameloblastoma, malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma; glioma, malignant; epithelioma; astrocytoma; protoplasmic astrocytoma;Fibrillary astrocytoma; astroblastoma; glioblastoma; anaplastic glioma; anaplastic astrocytoma; primitive neuroectodermal tumor; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor; meningioma, malignant; neurofibrosarcoma; schwannoma, malignant; granular cell tumor, malignant; malignant lymphoma; Hodgkin's disease; Hodgkin; paragranuloma; malignant lymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-Hodgkin lymphoma; B-cell lymphoma; low-grade / follicular non-Hodgkin 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-necrotizing cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; Waldenström macroglobulinemia; malignant histiocytosis; multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphocytic leukemia; plasma cell leukemia; erythroleukemia; lymphosarcoma cell leukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryocytic leukemia; myelosarcoma; hairy cell leukemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); acute myeloid leukemia (AML); and chronic myelogenous leukemia.;

[0345] The therapies provided herein may include the administration of combinations of therapeutic agents such as a first cancer therapeutic agent and a second cancer therapeutic agent. The therapies can be administered by any suitable method known in the art. For example, the first cancer therapy and the second cancer therapy can be administered sequentially (at different times) or simultaneously (at the same time). In some embodiments, the first and second cancer therapies are administered in separate compositions. In some embodiments, the first and second cancer therapies are in the same composition. Embodiments of the present disclosure relate to compositions and methods comprising therapeutic compositions. Different therapies can be administered in one composition or in two or more compositions such as two, three, or four compositions. Various combinations of agents can be employed. Examples of therapies other than those of the present disclosure include surgery, chemotherapy, drug therapy, radiation therapy, hormone therapy, immunotherapy (other than those of the present disclosure), or combinations thereof.

[0346] The therapies of the present disclosure may be administered by the same route of administration or by different routes of administration. In some embodiments, the cancer therapy can be administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by transplantation, by inhalation, intrathecally, intraventricularly, or nasally. In some embodiments, the antibiotic is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by transplantation, by inhalation, intrathecally, intraventricularly, or nasally. Appropriate dosages can be determined based on the type of disease being treated, the severity and course of the disease, the clinical condition of the individual, the individual's clinical history and response to treatment, and the discretion of the attending physician.

[0347] The treatment can include various "unit doses". A unit dose is defined as containing a predetermined amount of the therapeutic composition. The amount administered, as well as the particular route and formulation, are within the judgment of those skilled in the clinical arts. A unit dose need not be administered as a single injection, but may include continuous infusion over a period of time. In some embodiments, the unit dose includes a dose that can be administered in a single administration.

[0348] The dosage depends on the desired therapeutic effect, both in terms of the number of treatments and the unit dosage. The effective amount is understood to refer to the amount necessary to achieve a particular effect. In practice, in certain embodiments, it is contemplated that dosages in the range of 10 mg / kg to 200 mg / kg may affect the protective ability of these agents. Thus, dosages may include, for example, 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 μg / kg, mg / kg, μg / day, or mg / day, or any range derivable therefrom. Furthermore, such dosages can be administered multiple times during a day and / or over multiple days, weeks, or months.

[0349] In certain embodiments, an effective amount of the pharmaceutical composition can provide blood levels of from about 1 μM to 150 μM. In another embodiment, the effective amount is from about 4 μM to 100 μM; or from about 1 μM to 100 μM; or from about 1 μM to 50 μM; or from about 1 μM to 40 μM; or from about 1 μM to 30 μM; or from about 1 μM to 20 μM; or from about 1 μM to 10 μM; or from about 10 μM to 150 μM; or from about 10 μM to 100 μM; or from about 10 μM to about 50 μM; or from about 25 μM to about 150 μM; or from about 25 μM to about 100 μM; or from about 25 μM to about 50 μM; or from about 50 μM to about 150 μM; or from about 50 μM to about 100 μM (or any range derivable therefrom) of blood levels. In other embodiments, the dosage results in the following blood levels of the agent resulting from the therapeutic agent administered to the subject: about, at least, 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 μM, or any range derivable therefrom. In certain embodiments, the therapeutic agent administered to the subject is metabolized in the body to a metabolite therapeutic agent, in which case the blood concentration can refer to the amount of that therapeutic agent. Alternatively, to the extent the therapeutic agent is not metabolized by the subject, the blood concentrations discussed herein may refer to the unmetabolized therapeutic agent.

[0350] The exact amount of the therapeutic composition also depends on the judgment of the practitioner and is specific to each individual. Factors affecting dosage include the physical and clinical condition of the patient, the route of administration, the intended treatment goal (symptom relief versus cure), the efficacy, stability and toxicity of the particular therapeutic substance or other therapies the subject may be receiving.

[0351] It is understood and recognized by those skilled in the art that a dosage unit of μg / kg body weight or mg / kg body weight can be expressed by conversion into an equivalent concentration unit of μg / ml or mM (blood concentration) such as 4 μM to 100 μM. Also, it is understood that uptake depends on the species and organ / tissue. The conversion factors and physiological assumptions applicable to uptake and concentration measurements are well known, and those skilled in the art can convert one concentration measurement value to another and make reasonable comparisons and conclusions regarding the dosages, potencies, and results described herein.

[0352] V. Kit Certain embodiments of the present disclosure also relate to kits comprising the compositions of the present invention or compositions for practicing the methods of the present invention. In certain embodiments, the kit comprises NK cells that are fresh or frozen and that can or cannot be pre-activated or expanded. The NK cells can or cannot already express one or more components of the TCR / CD3 complex. If the NK cells do not yet express one or more components of the TCR / CD3 complex, the kit can include reagents for the corresponding transfection or transduction of the NK cells, such as vectors that express the component(s), primers for amplifying the component(s), and the like. In some cases, the NK cells can also or cannot express one or more heterologous proteins as defined herein, and if they do not express them, the kit can include vectors that express the heterologous protein(s), primers for amplifying the heterologous protein(s), and the like.

[0353] The kit can include components that are individually packaged or can be placed in a container such as a tube, bottle, vial, syringe, or other suitable container means. The individual components can also be provided in the kit in a concentrated amount. In some embodiments, the components are provided individually at the same concentration as in solution with the other components. The concentration of the components can be provided as 1×, 2×, 5×, 10×, or 20× or more.

Examples

[0354] VI. Examples The following examples are included to illustrate preferred embodiments of the present invention. The techniques disclosed in the examples that follow represent techniques that the inventors have found to function well in the practice of the present invention and, accordingly, can be considered to constitute preferred modes for its practice, which should be understood by those skilled in the art. However, those skilled in the art will appreciate that, in light of the present disclosure, many modifications can be made to the specific embodiments disclosed and still obtain a like or similar result without departing from the spirit and scope of the present invention.

[0355] Example 1 Preparation and effective use of NK cells expressing CD3 This example relates to a cancer immunotherapeutic agent as a strategy for redirecting the specificity of NK cells to one or more target antigens by "arming" or pre-complexing NK cells with bispecific or multispecific antibodies, which can be done, for example, prior to injection or by co-injecting the two products separately. The NK cells are transduced with one or more CD3 chains including the CD3ζ, CD3γ, CD3δ, and CD3ε chains and can be from any source. The cells can or cannot be expanded, can be pre-activated with one or more inflammatory cytokines such as IL12 / 15 / 18, and / or can be genetically modified to express engineered antigen receptors such as chimeric antigen receptors or TCRs, and / or cytokine genes, and / or one or more heterologous proteins such as homing / chemokine receptors.

[0356] Figures 1A and 1B show different embodiments of NK cells engineered to be utilized with bispecific or multispecific antibodies. As shown in Figure 1A, in first-generation NK cells, the cells are engineered to express CD3 such that they can be activated with a bispecific antibody or multispecific antibody, including a bispecific T cell engager (BiTE) that binds to a heterologous CD3 expressed on the surface of the NK cell. In another embodiment, NK cells expressing CD3 can be bound by a BiTE containing an anti-CD3 antibody, and the NK cells also express one or more specific cytokines (e.g., IL-15 and / or IL-21), resulting in increased efficacy and potency that is particularly useful for the treatment of solid tumors. In another embodiment, the NK cells are engineered to express CD3 such that they can be activated by a BiTE containing an anti-CD3 antibody and are also utilized with a bispecific antibody or multispecific antibody (e.g., a bispecific NK cell engager, or BiKE) that includes an antibody that binds to a surface antigen naturally present on NK cells, such as, for example, CD16, CS1, CD56, NKG2D, NKG2C, DNAM, 2B4, CD2, NCR, or KIR. In this way, the NK cells respond to both an NK engager and a T cell engager. In another embodiment, in addition to expressing CD3 to engage a T cell engager, the NK cells also express an engineered antigen receptor, such as a CAR or an engineered TCR.

[0357] Figure 1B shows different embodiments in which NK cells are modified to express both CD3 and TCR. On the right side, a T cell TCR with an α-chain and a β-chain with antigen-binding sites is shown, and the TCR complexes with CD3ζ to effect signal transduction. The T cell TCR is co-complexed with two CD3ε chains, a CD3δ chain, and a CD3γ chain. In some embodiments, the NK cell expresses a TCR in which one or more of the cytoplasmic domains of any of the CD3 molecules are heterologous intracellular domains such as those derived from CD16, NKG2D, DAP10, DAP12, NCR, and DNAM-1. As shown on the left side of Figure 1B, the NK cell is configured to express a CD3 co-receptor component, and in one example the CD3 component is CD3ε. In such a case, a standard BiTE (upper left, including an antibody against a tumor antigen and an antibody against CD3) that is normally utilized with T cells that naturally express CD3 can be utilized with NK cells that express CD3. In this particular example, the NK cell expresses a polypeptide that includes the extracellular domain of CD3ε (although the extracellular domains of other CD3 components can be utilized), and the extracellular domain of CD3ε is linked to the transmembrane domain and / or cytoplasmic domain of another molecule, such as the transmembrane domain and / or cytoplasmic domain of CD3ζ, CD16, NKG2D, DAP10, DAP12, NCR, or DNAM-1.

[0358] As described above, Figure 1C schematically shows the generation of a surface-expressible single chimeric CD3 construct that can be used with an anti-CD3 BiTE. As an example, the CD3 epsilon extracellular domain (ECD) is fused to the transmembrane domain (TM) of CD28, CD16, or NKG2D, with or without the CD3 zeta and / or DAP10 intracellular domain, and the intracellular domain (ICD) of CD28, CD16, or NKG2D. In one example, the construct is incorporated within the SFG retroviral vector backbone derived from Moloney murine virus, which can be used with a packaging plasmid for virus production. Thus, when a CD3-BiTE is used with such a construct of Figure 1C, the antibody binds to the extracellular domain of CD3ε.

[0359] Embodiments of the present disclosure utilize some or all of the CD3 receptor complex. As shown in FIGS. 2A and 2B, NK cells can be transfected or transduced with full-length CD3zeta, CD3gamma, CD3delta, and CD3epsilon. In such cases, the full length of each of CD3zeta, CD3gamma, CD3delta, CD3epsilon includes an extracellular domain, a transmembrane domain, and an intracellular domain. When different components of the receptor are expressed from the same vector, they can be configured to be produced as separate polypeptides, such as by utilizing an IRES or 2A element. In any case, any expression construct can be configured to express one or more cytokines including at least IL-15.

[0360] Figure 4 shows CD3 expression in NK cells on day 4 after CMV TCR complex transduction. This figure provides a FACS plot showing CD3 expression in NK cells 4 days after CMV TCR complex transduction. Non-transduced (NT) NK cells (CD56+ CD3-) function as a negative control, and T cells (CD3+ CD56-) function as a positive control.

[0361] Figure 5 shows TCR expression on NK cells on day 4 after CMV TCR complex transduction of NK. Specifically, it provides a FACS plot showing TCRa / b expression on NK cells 4 days after CMV TCR complex transduction. Non-transduced (NT) NK cells (CD56+ CD3-TCRa / b-) function as a negative control, and T cells (CD3+TCRa / b+ CD56-) function as a positive control.

[0362] Figure 6 shows TCR / CD3 expression on NK on day 6 after CMV TCR complex transduction. Specifically, the FACS plot shows dual expression of CD3 and TCRa / b on NK cells 6 days after CMV TCR complex transduction. Non-transduced (NT) NK cells (CD56+ CD3-TCRa / b-) function as a negative control, and T cells (CD3+TCRa / b+ CD56-) function as a positive control.

[0363] Figure 7 shows the binding of CD3-CD19 BiTE on NK cells via different concentrations of CD3 / TCR. Specifically, various cells (non-transduced (NT) NK cells, T cells, or three different NK-TCR cells) were incubated with blinatumomab, a CD3-CD19 bispecific engager (BiTe), at 37°C for 1 hour using two different concentrations (0.5 μg / μl or 4 μg / μl). Next, biotin-labeled CD19 antigen (CD19 CAR detection reagent from Miltenyi Biotech™) was added for 20 minutes, followed by addition of anti-biotin antibody for 15 minutes at room temperature. Using this strategy, any BiTe engaged with CD3+ cells was detected. The histograms in Figure 7 show the levels of CD19 binding to the CD3-CD19 bispecific engager (BiTe) that correlate with CD3 expression on NK-TCR and T cells.

[0364] Figure 8 shows NK-TCR cytokine production after stimulation with plate-bound CD3 antibody. In particular, CD3-OKT3 clone 20 μg / ml was incubated overnight at 4°C in flat-bottom 96-well plates to form plate-bound antigen. The next day, T cells or NK cells (NT or TCR-transduced) were added to the wells for 4 hours, and brefeldin A (which inhibits cytokine release and traps it in the cytoplasm for detection by intracellular cytokine staining) was added. Next, they were harvested for surface and intracellular staining, and cytokine production and degranulation (TNFα and CD107a) were evaluated. The FACS plots in Figure 8 show the TNFα and CD107a double-positive population in NK cells transduced with TCR. Non-transduced (NT) NK cells (CD56+ CD3-) function as a negative control, and T cells (CD3+ CD56-) function as a positive control.

[0365] Figure 9 shows the phosphorylation of CD3ζ in NK TCR / CD3 cells after crosslinking CD3. The various cells tested included non-transduced (NT) NK cells; non-transduced (NT) T cells, or NK cells transduced with three different CD3-TCRs (CD1, CD2, or CD3 represent different donors). Each NK cell group was transduced with CD3ZFLGDEFL15 (see FIGS. 2A and 2B). NK cells were incubated on ice for 20 minutes with CD3 OKT3 clone (Miltenyi, 130-093-387) at a concentration of 20 μg / ml. Next, the cells were crosslinked with Fab2 IgG1 antibody at various time points and stained to confirm CD3z phosphorylation. This analysis of CD3ζ is useful because, as an internalization signal from the surface, NK cells can be crosslinked with CD3 monoclonal antibody only if they express it. NK cells not transduced with CD3 show no phosphorylation or activation after stimulation.

[0366] NK cells transduced with CD3-TCR also increased with CD3 OKT3 stimulation and showed sustained signaling at basal levels similar to T cells, while non-transduced NK cells showed no CD3ζ phosphorylation at basal levels or with CD3 OKT3 stimulation.

[0367] Figure 10 shows that pre-culturing CD3-CD19 BiTE with NK cells expressing TCR / CD3 increases the killing activity against Raji cells. The NK cells were transduced with either CD3-TCR#1 (CD3ZFLGDEFL15 (see FIGS. 2A and 2B)) having a CD8 transmembrane domain for membrane-bound IL21 and containing full-length CD3ζ, full-length CD3γ, full-length CD3δ, and full-length CD3ε, or CD3-TCR#2 (also called Z2, CD3ZGDEFL8SP21CD8) which is linked to membrane-bound IL21. NK cells transduced with the CD3 / TCR construct or non-transduced NK cells were loaded with blinatumomab, incubated for 1 hour, and washed with PBS. Next, they were co-cultured with CD19+ B cell lymphoma cells at different effector cell:target cell ratios (FIG. 10A is a 1:1 ratio and FIG. 10B is a 1:5 ratio) over various time points. As used herein, the effector cells are CD-3-TCR NK cells and the target cells are Raji cells. NK cells transduced with CD3-TCR loaded with blinatumomab showed enhanced anti-tumor activity compared to non-transduced NK cells loaded with blinatumomab or NK cells transduced with CD3 / TCR, but were not loaded with blinatumomab at any E:T ratio.

[0368] Example 2 NY-ESO TCR in NK cells This example relates to the generation and use of NY-ESO TCR in NK cells. An example for the generation of the cells is shown in FIG. 11. This schematic shows one case where NK cells are first transduced with a uTNK15 construct incorporating the CD3 complex, NK co-stimulatory molecules and the signaling domain from IL-15, followed by a second transduction step to introduce the TCR molecule, thus generating NK cells that co-express the CD3 and NK signaling molecules, IL-15, and the TCR complex. In one embodiment, the NK cells are derived from cord blood and are expanded in complete medium using irradiated (100 Gy) universal antigen-presenting cell (uAPC) feeder cells (2:1 feeder cell:NK ratio) and recombinant human IL-2 (200 U / ml). To generate universal T cell-like NK cells (uTNK15 cells) capable of secreting IL-15, NK cells were purified and transduced 4 days after isolation with a retroviral construct containing the CD3 complex with NK co-stimulatory molecul...

Claims

1. a) A composition comprising engineered NK cells modified to express one or more transgenic polynucleotides encoding: b) Optionally, at least one cytokine, and c) At least one engineered TCR comprising one or more of TCR α chain, TCR β chain, TCR γ chain, and / or TCR δ chain, wherein the engineered TCR targets a KRAS antigen.

2. The composition of claim 1, wherein the NK cells are modified to express the TCR αβ chain or the TCR γδ chain.

3. The composition of claim 1, wherein the NK cells are modified to express CD3ζ, two of CD3ε, CD3δ, and a part or all of CD3γ.

4. The composition of claim 1, wherein the NK cells are modified to express the full length of CD3ζ, CD3ε, CD3δ, and / or CD3γ.

5. The composition of claim 1, wherein any one or more of CD3ζ, CD3ε, CD3δ, and CD3γ are heterologously linked to one or more intracellular signaling domains.

6. The composition of claim 5, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and combinations thereof.

7. The composition of claim 6, wherein the intracellular signaling domain comprises a DAP10 intracellular signaling domain.

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

115.

9. The composition of claim 7, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

115.

10. The composition of claim 6, wherein the intracellular signaling domain comprises a CD28 intracellular signaling domain.

11. The composition of claim 10, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

116.

12. The composition of claim 10, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

116. ​

13. The composition according to claim 6, wherein the intracellular signaling domain comprises the DAP10 and CD28 intracellular signaling domains.

14. The composition according to claim 13, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

117.

15. The composition according to claim 13, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

117.

16. The composition according to claim 1, further comprising one or more monospecific antibodies, bispecific antibodies, or multispecific antibodies, wherein the bispecific antibody or multispecific antibody comprises an anti-CD3 antibody.

17. The composition according to claim 16, wherein the NK cells express the antibody.

18. The composition according to claim 16, wherein the NK cells are complexed with the antibody.

19. The composition according to claim 16, wherein the antibody is imigatuzumab, amivantamab, and / or cetuximab.

20. The composition according to claim 1, wherein the NK cells are derived from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or a mixture thereof.

21. The composition according to claim 1, wherein the NK cells are pre-activated.

22. The composition according to claim 21, wherein the NK cells are pre-activated with one or more cytokines.

23. The composition according to claim 22, wherein the cytokine is IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, or a combination thereof.

24. The composition according to claim 1, wherein the NK cells are expanded.

25. The composition according to claim 24, wherein the NK cells are expanded in the presence of IL-2.

26. The composition according to claim 1, wherein the NK cells are modified to express one or more additional heterologous proteins.

27. The composition according to claim 26, wherein the additional heterologous protein is an engineered antigen receptor, cytokine, homing receptor, and / or chemokine receptor.

28. The composition according to claim 1, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 299 and a sequence that is at least 85% identical to SEQ ID NO:

301.

29. The composition according to claim 1, wherein the TCR comprises one or more sequences that are at least 85% identical to SEQ ID NOs: 299-317.

30. The composition according to claim 1, wherein the target KRAS antigen epitope comprises and / or consists of GADGVGKSAL (SEQ ID NO: 292) and / or GADGVGKSAL (SEQ ID NO: 293).

31. The composition according to claim 30, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 304 and a sequence that is at least 85% identical to SEQ ID NO:

305.

32. The composition according to claim 30, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 306 and a sequence that is at least 85% identical to SEQ ID NO:

307.

33. The composition according to claim 30, wherein the TCR comprises a sequence that is at least 85% identical to SEQ ID NO: 308 and a sequence that is at least 85% identical to SEQ ID NO:

309.

34. The composition according to claim 27, wherein the heterologous protein is a cytokine.

35. The composition according to claim 34, wherein the cytokine is selected from the group consisting of IL-15, IL-12, IL-2, IL-18, IL-21, IL-23, IL-7, GMCSF, or a combination thereof.

36. The composition according to claim 35, wherein the cytokine is membrane-bound.

37. The composition according to claim 35, wherein the cytokine is IL-15.

38. The composition according to claim 36, wherein the membrane-bound cytokine comprises a transmembrane domain of CD8, CD28, CD27, B7H3, IgG1, IgG4, CD4, DAP10, or DAP12.

39. The composition according to claim 27, wherein the NK cells express a chimeric antigen receptor and a cytokine.

40. The composition according to claim 16, wherein the monospecific antibody, bispecific antibody, or multispecific antibody comprises an antibody that targets a cancer antigen.

41. The composition according to claim 40, wherein the cancer antigen is an EGFR, C-met, and / or TROP-2 antigen.

42. a) NK cells modified to express some or all of the CD3 receptor complex and modified to express a T cell receptor (TCR) αβ chain or TCR γδ chain; and b) a monospecific antibody, bispecific antibody, or multispecific antibody, wherein the bispecific antibody or multispecific antibody comprises an anti-CD3 antibody that binds to CD3 on the NK cells, the monospecific antibody, bispecific antibody, or multispecific antibody A composition comprising a complex comprising a composition wherein the TCR targets a KRAS antigen. **Claim 43** The composition according to claim 42, wherein the NK cells are modified to express a TCR αβ chain that is at least 85% identical to SEQ ID NO: 299 and SEQ ID NO: 301, and the antibody is inotuzumab, amivatumab, and / or cetuximab. **Claim 44** The composition according to claim 43, wherein the NK cells are modified to express full-length CD3ζ, CD3ε, CD3δ, and / or CD3γ. **Claim 45** The composition according to claim 44, wherein any one or more of CD3ζ, CD3ε, CD3δ, and CD3γ are heterologously linked to one or more intracellular signaling domains. **Claim 46** The composition according to claim 45, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, DNAM, and combinations thereof. **Claim 47** The composition according to claim 46, wherein the intracellular signaling domain comprises a DAP10 intracellular signaling domain. **Claim 48** The composition according to claim 47, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

115. **Claim 49** The composition according to claim 47, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

115. **Claim 50** The composition according to claim 46, wherein the intracellular signaling domain comprises a CD28 intracellular signaling domain. **Claim 51** The composition according to claim 50, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

116. **Claim 52** The composition according to claim 50, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

116. **Claim 53** The composition according to claim 46, wherein the intracellular signaling domain comprises DAP10 and CD28 intracellular signaling domains. **Claim 54** The composition according to claim 53, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

117. **Claim 55** The composition according to claim 53, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

117. **Claim 56** The composition according to claim 42, wherein the complex is contained in a pharmaceutically acceptable excipient.

57. The composition according to claim 56, wherein the complex is contained in a delivery device.

58. A method of treating cancer in an individual, comprising administering to the individual a therapeutically effective amount of any one of the compositions according to claims 1 to 57.

59. A method of treating cancer in an individual, comprising administering to the individual a therapeutically effective amount of any one of the compositions according to claims 16, 17, 18, 19, 40, 41, or 42 to 57, wherein NK cells and an antibody are administered to the individual simultaneously, and optionally, the NK cells and the antibody are administered in the same formulation and / or pre-complexed before being administered to the individual.

60. The method according to claim 58, wherein the NK cells and the antibody are administered to the individual at different times.

61. The method according to claim 59, wherein the NK cells and the antibody are administered by injection.

62. The method according to claim 59, wherein the NK cells are autologous with respect to the individual.

63. The method according to claim 59, wherein the NK cells are allogeneic with respect to the individual.

64. A method of redirecting the specificity of NK cells against a cancer antigen for the treatment of an individual with a monospecific antibody, bispecific antibody or multispecific antibody, wherein the bispecific antibody or multispecific antibody comprises an anti-CD3 antibody, and administering to the individual the antibody and NK cells that express a part or all of the CD3 receptor complex and a part or all of the TCRαβ chain or TCRγδ chain. The method, wherein the TCR targets the KRAS antigen.

65. The method according to claim 64, wherein the NK cells are modified to express full-length CD3ζ, CD3ε, CD3δ, and / or CD3γ.

66. The method according to claim 65, wherein any one or more of CD3ζ, CD3ε, CD3δ, and CD3γ are heterologously linked to one or more intracellular signaling domains.

67. The method according to claim 66, wherein the intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, DNAM, and combinations thereof.

68. The method according to claim 67, wherein the intracellular signaling domain comprises the DAP10 intracellular signaling domain. **Claim 69** The method according to claim 68, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

115. **Claim 70** The method according to claim 68, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

115. **Claim 71** The method according to claim 67, wherein the intracellular signaling domain comprises the CD28 intracellular signaling domain. **Claim 72** The method according to claim 71, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

116. **Claim 73** The method according to claim 71, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

116. **Claim 74** The method according to claim 67, wherein the intracellular signaling domain comprises the DAP10 and CD28 intracellular signaling domains. **Claim 75** The method according to claim 74, wherein the intracellular signaling domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:

117. **Claim 76** The method according to claim 74, wherein the intracellular signaling domain comprises the amino acid sequence according to SEQ ID NO:

117. **Claim 77** The method according to claim 64, wherein the TCR chain is a TCR αβ chain and is at least 85% identical to SEQ ID NO: 299 and SEQ ID NO:

301. **Claim 78** The method according to claim 64, wherein the TCR chain is a TCR αβ chain and is at least 85% identical to one or more of SEQ ID NOs: 299-317. **Claim 79** The method according to claim 64, wherein the target KRAS antigen epitope comprises and / or consists of GADGVGKS A (SEQ ID NO: 293) and / or GADGVGKS AL (SEQ ID NO: 292). **Claim 80** The method according to claim 78, wherein the TCR chain comprises a sequence that is at least 85% identical to SEQ ID NO: 304 and a sequence that is at least 85% identical to SEQ ID NO:

305. **Claim 81** The method according to claim 78, wherein the TCR chain comprises a sequence that is at least 85% identical to SEQ ID NO: 306 and a sequence that is at least 85% identical to SEQ ID NO:

307. **Claim 82** The method according to claim 78, wherein the TCR chain comprises a sequence that is at least 85% identical to SEQ ID NO: 308 and a sequence that is at least 85% identical to SEQ ID NO:

309. **Claim 83** The method according to claim 64, further comprising the step of modifying the NK cells to express one or more additional heterologous proteins.

Images