Multi-receptor natural killer cells

Abstract

Embodiments of the disclosure include methods and compositions in which NK cells are modified by the hand of man to express a heterologous a) polypeptide comprising a CD16 Fc binding domain, b) a T Cell Receptor (TCR, e.g., an invariant T Cell Receptor (iTCR), and c) a CD3 co-receptor on NK cells that do not naturally express them. Such modified NK cells work effectively with monospecific, bispecific, or multi-specific antibodies, that may have modified or non-modified Fc domains.

Description

MULTI-RECEPTOR NATURAL KILLER CELLSCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of U. S. Provisional Patent Application 63 / 500,423 filed on May 5, 2023, and U.S. Provisional Patent Application 63 / 601,150 filed on November 20, 2023, the contents of each of which are hereby incorporated by reference in their entirety.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted in ST26 format and is hereby incorporated by reference in its entirety. Said ST26 copy, created on May 2, 2023, is named MDAC_1358_Sequence_Listing.xml and is 256,797 bytes in size.TECHNICAL FIELD AND BACKGROUNDI. Technical Field

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

[0004] Natural killer (NK) cells have been studied as potential anti-tumor effectors, yet a number of barriers limit their therapeutic exploitation, mainly related to their lack of antigen specificity. One approach to overcome this is to transduce NK cells with a chimeric antigen receptor (CAR) or an engineered T-cell receptor (TCR) to target a desired antigen. In T cells, one can utilize a bispecific or multi-specific antibody, such as a bispecific T cell engager (BiTE) that binds CD3 on the surface of T cells and that also binds an antigen on the surface of target cells (e.g., cancer cells). CD3 is composed of four distinct chains, and in mammals, the complex contains a CD3y chain, a CD35 chain, and two CD3s chains. These chains associate with the T-cell receptor (TCR) and the (^-chain (zeta-chain) to generate an activation signal in T lymphocytes. However, NK cells do not naturally express the CD3 receptor complex or TCRs, and thus are not effectively utilized in conjunction with BiTEs. While NK cells naturally express FcyR proteins such as FcyRIII (CD16), polymorphisms in the extracellular Fc recognition domain can result in lack of (or poor) binding of non-Fc domain modified (e.g., glycoengineered) antibodies to NK cell surfaces.

[0005] The present disclosure satisfies a long-felt need in the art to improve upon the effectiveness, safety profile, & targeting capacity of immunotherapies, including those that utilize NK cells.SUMMARY

[0006] Autologous anti-CD19 chimeric antigen receptor (CAR) T-cells have been shown to induce remissions in 57-71% of patients with chronic lymphocytic leukemia (CLL), 52-82% of patients with diffuse large B-cell lymphoma (DLBCL), and 78-92% of patients with low grade non-Hodgkin lymphoma (LG-NHL). Indeed, there are currently multiple FDA-approved autologous anti-CD19 CAR T-cell products available for clinical use. However, CAR T-cells have recognized limitations including the cost of therapy and the time required to collect the T-cells and manufacture the product. Moreover, a proportion of patients treated with CAR T- cells develop toxicities such as cytokine release syndrome (CRS), neurotoxicity, or hemophagocytic lymphohistiocytosis (HLH), each of which carry significant morbidity. These limitations require CAR-T cells to be administered by specialized teams, further limiting access to these life-saving-therapies. Therefore, there is great interest in developing off-the-shelf cell therapies that are cost effective, safe and potent. Natural killer (NK) cells target cancer cells that downregulate HLA class-I molecules or express stress markers in a non-antigen specific manner, thus playing a critical role in cancer immunevigilance. These cells can be engineered to express various transgenes, and can be safely administered without the need for HLA- matching, thus, eliminating the need to produce an immunotherapy product on an individual patient basis. This property makes NK cells especially attractive for off-the-shelf therapy, thus, reducing the cost of manufacturing and expanding access to these potentially life-saving therapies to many more patients.

[0007] Embodiments of the disclosure include methods and compositions for treatment of an individual with a disorder, such as but not limited to an autoimmune disorder, cancer, and / or an infection, using adoptive cell therapy. In specific embodiments, the individual is provided a therapeutically effective amount of a bipartite therapy that includes a modified immune cell (e.g., a modified NK cell) and antibodies that are capable of being able to bind the NK cells to initiate signaling, activation, and / or killing of target cells. The disclosure concerns NK cells that have been modified to express multiple proteins that are not naturally expressed in NK cells and that work in conjunction together, including heterologous proteins on the surface of the NK cells that are naturally not present in NK cells. The disclosure also concerns NK cells that have been modified to overexpress proteins that do naturally occur in NK cells.

[0008] Provided herein are polynucleotides, which can comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to transcriptional reading frames comprising SEQ ID NO: 171-175. A polynucleotide encoding a sequence of interested can be comprised in a vector. A vector can comprise at least about 80%, 85%, 90%, 95%, 98%, or 100% sequence identity to SEQ ID NOs: 177-181.

[0009] Provided herein are polynucleotides comprising a sequence encoding a T cell receptor (TCR) alpha and TCR beta polypeptide, and / or TCR gamma and TCR delta polypeptide, and a polypeptide comprising a CD16 derived Fc binding domain. TCR polypeptides can be invariant TCR (iTCR) polypeptides. iTCRa and / or iTCRp polypeptides and / or polynucleotides encoding the same can comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 51-149. An iTCRp polypeptide can comprise a polynucleotide encoding a VP-DJ region that is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 75-149. An iTCRp polypeptide and / or polynucleotide encoding the same can comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NO: NOs: 59-74. An encoded iTCRp polypeptide can be at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 60. A polynucleotide encoding an iTCRp polypeptide can be at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 59. An iTCRa polypeptide and / or polynucleotide encoding the same can comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NOs: 51-52.

[0010] A polynucleotide comprising a sequence encoding a CD16 derived Fc binding domain can comprise a coding sequence for a human CD16A Fc binding domain. A polynucleotide encoding a polypeptide comprising a CD16 derived Fc binding domain can comprise the Fc binding domain being fused in N to C terminus order to an optional hinge domain, a transmembrane domain (TMD), and one or more optional intracellular signaling domains (ICD). A TMD can be derived from CD16, or CD3(^. A TMD can comprise or consist of a sequence with at least about 90% identity SEQ ID NOs: 163 or 167. A hinge domain can be derived from CD32. In certain cases, a hinge domain can be a hinge domain as previously described in the art. A hinge domain can comprise or consist of a sequence with at least about 90% identity to SEQ ID NO: 161. An ICD can be derived from CD16 and / or CD3(^. A polypeptide can comprise a CD 16 derived Fc binding domain that does not comprise a mutation that renders the CD16 derived Fc binding domain resistant to cleavage. A coding sequence for a polypeptide comprising a CD 16 derived Fc domain can be at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 150-154. A polynucleotideprovided herein can comprise a sequence encoding one or more cytokine sequences. A cytokine can comprise IL- 15 and / or IL-21.

[0011] Also provided herein are engineered NK cells comprising polynucleotides disclosed herein. Additionally, provided herein are methods of treating a disease in an individual, said method comprising administering to an individual in need thereof an engineered NK cell comprising one or more polynucleotides disclosed herein.

[0012] Provided herein are engineered immune cells comprising one or more transgenic polynucleotides, where the transgenic polynucleotides encode: a) a CD3 protein complex comprising part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3^, b) optionally at least one cytokine, c) at least one TCRa and TCRP chain and / or a TCRy and TCR5 chain, and d) a polypeptide comprising a CD16 Fc binding domain. One or more transgenic polynucleotides can comprise multi ci str onic transcriptional open reading frames. The NK cells can be modified to express part or all of CD36, two of CD3s, CD3y, and / or CD3^. One or more of CD36, CD3s, CD3y, and / or CD3<^ can be linked to one or more heterologous intracellular signaling domains. A heterologous intracellular signaling domain can be selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and a combination thereof. A heterologous intracellular signaling domain can comprise a DAP 10 intracellular signaling domain. A heterologous intracellular signaling domain can comprise an amino acid sequence at least about 85% identical to SEQ ID NO: 42. A heterologous intracellular signaling domain can comprise a CD28 intracellular signaling domain. A heterologous intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 43. A heterologous intracellular signaling domain can comprise a DAP 10 and CD28 intracellular signaling domain. A heterologous intracellular signaling domain can comprise an amino acid sequence at least about 85% identical to SEQ ID NO: 44.

[0013] An engineered immune cell provided herein can comprise coding sequences for a CD3 protein complex and at least one cytokine that can be comprised in a first multi ci str onic construct, wherein coding sequences for at least one TCRa and TCRP chain and / or a TCRy and TCR5 chain, and the polypeptide comprising a CD16 Fc binding domain can also be encoded by a second multi ci str onic construct. An engineered immune cell can include a coding sequence for a cytokine, wherein the cytokine comprises IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, IL-23, and / or GMCSF. An engineered immune cell can be modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 45), UT-NK15-28 (SEQ ID NO: 47), or UTNK15-28-DAP10 (SEQ ID NO: 49). A cytokine can comprise IL-15 and / orIL-21. A cytokine can comprise IL-15 and can comprise a polypeptide sequence and / or polynucleotide sequence encoding the same that is at least 85% identical to any one or more of SEQ ID NOs: 182-183. A cytokine can comprise IL-21 and can comprise a polypeptide sequence and / or polynucleotide sequence encoding the same that is at least 85% identical to any one or more of SEQ ID NOs: 184-187.

[0014] An engineered immune cell provided herein can comprise a TCR, where the TCR polypeptides are invariant TCR (iTCR) polypeptides. An iTCRa and iTCRP polypeptides and / or polynucleotides encoding the same can comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 51-149. An iTCRP polypeptide can comprise a polynucleotide encoding a VP-DJ region that is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 75-149. An iTCRP polypeptide and / or polynucleotide encoding the same can comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NO: NOs: 59-74. An encoded iTCRP polypeptide can be at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 60. A polynucleotide encoding the iTCRP polypeptide can be at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 59. An iTCRa polypeptide and / or polynucleotide encoding the same can comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 51-52.

[0015] An engineered immune cell provided herein can comprise a polypeptide comprising a CD16 Fc binding domain (e.g., a CD16 extracellular domain), wherein the CD16 Fc binding domain can comprise a human CD16 Fc binding domain. A polypeptide comprising a CD16 derived Fc binding domain can comprise a human CD16A Fc binding domain. A polypeptide comprising a CD 16 Fc binding domain can be fused (in N to C terminus order) to an optional hinge domain, a transmembrane domain (TMD), and one or more optional intracellular signaling domains (ICD). A TMD can be derived from CD 16, or CD3(^. A TMD can comprise or consist of a sequence with at least about 90% identity to SEQ ID NOs: 163 or 167. A hinge domain can be derived from CD32. A hinge domain can comprise or consist of a sequence with at least about 90% identity to SEQ ID NO: 161. An ICD can be derived from CD16 and / or CD3(^. An engineered immune cell provided herein can comprise a coding sequence that is least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 171- 175.

[0016] An engineered immune cell provided herein can be a Natural Killer (NK) cell. An NK cell can be derived from cord blood (CB), peripheral blood (PB), bone marrow, stem cells,or a combination thereof. An NK cell can be a primary NK cell, and can not be derived from stem cells and / or induced pluripotent stem cells (iPSCs).

[0017] Also provided herein are compositions comprising engineered NK cells loaded with (e.g., complexed with) one or more antibodies. One or more antibodies can be one or more monospecific, bispecific, or multi-specific antibodies. At least one or more antibodies can comprise a glycoengineered Fc domain that has a high affinity to wild type CD16. At least one or more antibodies can comprise a non-glycoengineered Fc domain that has a low affinity to wild type CD 16 Fc binding domains. A non-glycoengineered Fc domain can be loaded on (complexed to) the transgenic polypeptide comprising a CD16 Fc binding domain. One or more antibodies can comprise an IgGl and / or IgG4 Fc domain.

[0018] One or more antibodies target antigens can be CD3, CD16, CD28, EGFR, c-MET, CD30, PSMA, MUC17, CD33, FLT3, STEAP1, BCMA, CLDN18.2, CD123, CD19, CD20, EpCAM, CEA, GPC3, CD38, CD33, CD22, HER2, GPA33, GD2, MUC16, GPRC5D, DLL- 3, CLEC12A, and / or SSTR. One or more antibodies targets can be CD3, CD16, CD28, CD19, CD20, CD30, HER2, GPRC5D, EGFR, c-MET, FcRH5, and / or BCMA. One or more antibodies can comprise [fam]-trastuzumab deruxtecan, Abciximab, Adalimumab, Ado- trastuzumab emtansine, Aducanumab, Alemtuzumab, Alirocumab, Amivantamab, Anifrolumab, Ansuvimab, Atezolizumab, Atoltivimab with Maftivimab and Odesivimab-ebgn (aka Inmazeb), Avelumab, Basiliximab, Belantamab mafodotin, Belimumab, Benralizumab, Bevacizumab, Bezlotoxumab, Bimekizumab, Blinatumomab, Brentuximab vedotin, Brodalumab, Brolucizumab, Burosumab, Canakinumab, Caplacizumab, Casirivimab + imdevimab, Catumaxomab, Cemiplimab, Certolizumab pegol, Cetuximab, Cevostamab, Crizanlizumab, Daclizumab, Daratumumab, Denosumab, Dinutuximab, Donanemab, Dostarlimab, Dupilumab, Durvalumab, Eculizumab, Edrecolomab, Efalizumab, Elotuzumab, Emapalumab, Emicizumab, Enfortumab vedotin, Eptinezumab, Erenumab, Evinacumab, Evolocumab, Faricimab, Fremanezumab, Galcanezumab, Gemtuzumab, Gemtuzumab- Ozogamicin, Golimumab, Guselkumab, Ibalizumab, Ibritumomab tiuxetan, Idarucizumab, Imgatuzumab, Inebilizumab, Infliximab, Inolimomab, Inotuzumab, Inotuzumab-Ozogamicin, IPH61, Ipilimumab, Isatuximab, Ixekizumab, Lanadelumab, Lecanemab, Loncastuximab tesirine, Margetuximab, Mepolizumab, Mirvetuximab soravtansine, Mogamulizumab, Mosunetuzumab, Moxetumomab pasudotox, Murom onab-CD3, Narsoplimab, Natalizumab, Naxitamab, Nebacumab, Necitumumab, Nirsevimab, Nivolumab, Obiltoxaximab, Obinutuzumab, Ocrelizumab, Ofatumumab, Olaratumab, Omalizumab, Omburtamab, Oportuzumab monatox, Palivizumab, Panitumumab, Pembrolizumab, Penpulimab,Pertuzumab, Polatuzumab vedotin, Ramucirumab, Ranibizumab, Ravulizumab, Raxibacumab, Regdanvimab, Relatlimab, Reslizumab, Retifanlimab, Risankizumab, Rituximab, Romosozumab, Sacituzumab govitecan, Sarilumab, Satralizumab, Secukinumab, Siltuximab, Sintilimab, Sotrovimab, Spesolimab, Sutimlimab, Tafasitamab, Tebentafusp, Teclistamab, Teplizumab, Teprotumumab, Tezepelumab, Tildrakizumab, Tislelizumab, Tisotumab vedotin, Tixagevimab, cilgavimab, Tocilizumab, Toripalimab, Tositumomab-1131, Tralokinumab, Trastuzumab, Tremelimumab, Ublituximab, Ustekinumab, Vedolizumab, AMG 160 / Acapatamab, AMG 199 / TNB 585, AMG 330, AMG 427 / EMIRODATAMAB, AMG 509, AMG 701, AMG 910, APVO414 / ES414 / MOR209, APVO436, Catumaxomab / Removab, CC- 1, CC-93269 / EM801, Cibisatamab / RG7802 / RO6958688, CLN-049, Elranatamab / PF- 06863135, EMB-06, Epcoritamab / GEN3013, ERY974, Flotetuzumab / MGD006,Glofitamab / RG6026 / RO7082859, ISB 1342 / GBR 1342, JNJ-63709178, JNJ-63898081, JNJ- 67571244, JNJ-75348780, Linvoseltamab / REGN 5458, M701, M802, MGD007, Mosunetuzumab / RG7828, Nivatrotamab / Hu3F8-BsAb, Odronextamab / REGN1979, REGN4018, REGN5459, REGN7075, REGN5678, Talquetamab / JNJ-64407564,Tarlatamab / AMG 757, Tepoditamab / MCLA-117, TNB-383B, TNB-486, TNB-585, XmAbl3676 / Plamotamab, XmAbl4045 / Vibecotamab, XmAbl8087 / Tidutamab, and / or AFM13. One or more antibodies can comprise Elranatamab, Imgatuzumab, Margetuximab, Amivantamab, Blinatumomab, Obinutuzumab, IPH61 (also known as IPH6101 or SAR443579), Teclistamab, Cetuximab, Talquetamab, Pertuzumab, Trastuzumab, Tafasitamab, Brentuximab, and / or Rituximab. One or more antibodies can comprise or consist of Elranatamab. One or more antibodies can comprise or consist of Imgatuzumab. One or more antibodies can comprise or consist of Margetuximab. One or more antibodies can comprise or consist of Amivantamab. One or more antibodies can comprise or consist of Blinatumomab. One or more antibodies can comprise or consist of Obinutuzumab. One or more antibodies can comprise or consist of IPH61. One or more antibodies can comprise or consist of Teclistamab. One or more antibodies can comprise or consist of Cetuximab. One or more antibodies can comprise or consist of Rituximab. An NK cell can be engineered to express the one or more antibodies. One or more antibodies can comprise or consist of Talquetamab. One or more antibodies can comprise or consist of Pertuzumab. One or more antibodies can comprise or consist of Trastuzumab. One or more antibodies can comprise or consist of Tafasitamab. One or more antibodies can comprise or consist of Brentuximab.

[0019] An engineered NK cell can be modified to express one or more additional heterologous proteins selected from the group consisting of an antigen receptor, a cytokine, ahoming receptor, a chemokine receptor, and a combination thereof. An engineered NK cell can be pre-activated with one or more cytokines. A pre-activation cytokine can be IL-2, IL-7, IL- 12, IL-15, IL-18, IL-21, or any combination thereof. An engineered NK cell can comprise one or more engineered mutations in an endogenous gene. An engineered mutation in an endogenous gene can be a mutation in GR, TGFBR2, CISH, and / or CD38. An engineered NK cell provided herein can be comprised in a composition, optionally the composition can include a pharmaceutically acceptable excipient and / or be comprised in a delivery device.

[0020] Also provided herein are methods of treating a disease in an individual. Methods of treating an individual can comprise the step of administering to the individual a therapeutically effective amount of any one or more of the cells or compositions described herein. A disease can be an autoimmune disease, infection, and / or cancer. A disease can be cancer. A cancer can express tumor associated antigens (TAAs). A cancer can express CD19, CD20, CD30, HER2, GPRC5D, EGFR, c-MET, and / or BCMA. A cancer can be pancreatic cancer, colorectal cancer, ovarian cancer, kidney cancer, glioblastoma, breast cancer, renal cancer, myeloma, and / or leukemia. A method of treating an individual can further comprise administering to the individual at the same time or at different time, one or more monospecific, bispecific, and / or multispecific antibodies. One or more antibodies can comprise Elranatamab, Imgatuzumab, Margetuximab, Amivantamab, Blinatumomab, Obinutuzumab, IPH61 (also known as IPH6101 or SAR443579), Teclistamab, Cetuximab, and / or Rituximab. One or more antibodies can be administered at the same time as an engineered NK cell, and / or the one or more antibodies and the engineered NK cells can be complexed prior to administration to the individual. One or more antibodies can be administered prior to, after, and / or at the same time as an engineered NK cell. One or more antibodies can be administered more than one time prior to, after, and / or at the same time as an engineered NK cell.

[0021] Certain embodiments of the present invention are characterized through the following enumerated aspects.

[0022] Aspect 1, is a polynucleotide comprising a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to transcriptional reading frames comprising SEQ ID NO: 171- 175.

[0023] Aspect 2, is the polynucleotide of aspect 1, wherein the polynucleotide is comprised in a vector comprising at least about 80%, 85%, 90%, 95%, 98%, or 100% sequence identity to SEQ ID NOs: 177-181.

[0024] Aspect 3, is a polynucleotide comprising a sequence encoding a T cell receptor (TCR) alpha and TCR beta polypeptide, and / or TCR gamma and TCR delta polypeptide, and a polypeptide comprising a CD 16 derived Fc binding domain.

[0025] Aspect 4, is the polynucleotide of aspect 3, wherein the TCR polypeptides are invariant TCR (iTCR) polypeptides.

[0026] Aspect 5, is the polynucleotide of aspect 4, wherein the iTCRa and iTCRp polypeptides and / or polynucleotides encoding the same comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 51-149.

[0027] Aspect 6, is the polynucleotide of any one of aspects 3 to 5, wherein the iTCRp polypeptide comprises a polynucleotide encoding a VP-DJ region that is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 75-149.

[0028] Aspect 7, is the polynucleotide of any one of aspects 3 to 6, wherein the iTCRp polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NO: NOs: 59-74.

[0029] Aspect 8, is the polynucleotide of any one of aspects 3 to 7, wherein the encoded iTCRp polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 60.

[0030] Aspect 9, is the polynucleotide of aspect 8, wherein the polynucleotide encoding the iTCRp polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 59.

[0031] Aspect 10, is the polynucleotide of any one of aspects 3 to 9, wherein the iTCRa polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NOs: 51-52.

[0032] Aspect 11, is the polynucleotide of any one of aspects 3 to 10, wherein the polypeptide comprising a CD16 derived Fc binding domain comprises a human CD16A Fc binding domain.

[0033] Aspect 12, is the polynucleotide of any one of aspects 3 to 11, further comprising the Fc binding domain being fused in N to C terminus order to an optional hinge domain, a transmembrane domain (TMD), and one or more optional intracellular signaling domains (ICD).

[0034] Aspect 13, is the polynucleotide of aspect 12, wherein the TMD is derived from CD 16, or CD3<

[0035] Aspect 14, is the polynucleotide of aspect 12 or 13, wherein the TMD comprises or consists of a sequence with at least about 90% identity SEQ ID NOs: 163 or 167.

[0036] Aspect 15, is the polynucleotide of any one of aspects 12 to 14, wherein the hinge domain is derived from CD32.

[0037] Aspect 16, is the polynucleotide of any one of aspects 12 to 15, comprising a hinge domain that comprises or consists of a sequence with at least about 90% identity to SEQ ID NO: 161.

[0038] Aspect 17, is the polynucleotide of any one of aspects 12 to 16, comprising an ICD derived from CD 16 and / or CD3(^.

[0039] Aspect 18, is the polynucleotide of any one of aspects 3 to 17, wherein the polypeptide comprising a CD16 derived Fc binding domain does not comprise a mutation that renders the CD16 derived Fc binding domain resistant to cleavage.

[0040] Aspect 19, is the polynucleotide of any one of aspects 3 to 18, comprising a coding sequence that is least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 150-154.

[0041] Aspect 20, is the polynucleotide of any one of aspects 3 to 19, wherein the polynucleotide further encodes one or more cytokine sequences.

[0042] Aspect 21, is the polynucleotide of aspect 20, wherein the cytokine comprises IL- 15 and / or IL-21.

[0043] Aspect 22, is an engineered NK cell comprising the polynucleotide of any one of aspects 1-22.

[0044] Aspect 23, is a method of treating a disease in an individual, the method comprising administration of the engineered NK cells of aspect 22 to an individual in need thereof.

[0045] Aspect 24, is an engineered immune cell comprising, one or more transgenic polynucleotides encoding: a) a CD3 protein complex comprising part or all of a single chain or any combination of CD36, CD3s, CD3y, or CD3<^, b) optionally at least one cytokine, c) at least one TCRa and TCRP chain and / or a TCRy and TCR5 chain, and d) a polypeptide comprising a CD16 Fc binding domain.

[0046] Aspect 25, is the engineered immune cell of aspect 24, wherein the one or more transgenic polynucleotides comprise multicistronic transcriptional open reading frames.

[0047] Aspect 26, is the engineered immune cell of aspect 24 or 25, wherein the cells are modified to express part or all of CD36, two of CD3s, CD3y, and / or CD3^.

[0048] Aspect 27, is the engineered immune cell of any one of aspects 24 to 26, wherein any one or more of CD36, CD3s, CD3y, and / or CD3<^ are linked to one or more heterologous intracellular signaling domains.

[0049] Aspect 28, is the engineered immune cell of aspect 27, wherein the heterologous intracellular signaling domain is selected from the group consisting of CD 16, NKG2D, DAP 10, DAP12, 2B4, 4-1BB, CD2, CD28, and a combination thereof.

[0050] Aspect 29, is the engineered immune cell of any one of aspects 27 to 28, wherein the heterologous intracellular signaling domain comprises a DAP 10 intracellular signaling domain.

[0051] Aspect 30, is the engineered immune cell of aspect 29, wherein the heterologous intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 42.

[0052] Aspect 31, is the engineered immune cell of any one of aspects 27 to 30, wherein the heterologous intracellular signaling domain comprises a CD28 intracellular signaling domain.

[0053] Aspect 32, is the engineered immune cell of aspect 31, wherein the heterologous intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 43.

[0054] Aspect 33, is the engineered immune cell of any one of aspects 27 to 32, wherein the heterologous intracellular signaling domain comprises a DAP 10 and CD28 intracellular signaling domain.

[0055] Aspect 34, is the engineered immune cell of aspect 33, wherein the heterologous intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 44.

[0056] Aspect 35, is the engineered immune cell of any one of aspects 24 to 34, wherein coding sequences for a CD3 protein complex and at least one cytokine are comprised in a first multi ci str onic construct, and wherein coding sequences for at least one TCRa and TCRP chain and / or a TCRy and TCR5 chain, and the polypeptide comprising a CD16 Fc binding domain are encoded by a second multi ci str onic construct.

[0057] Aspect 36, is the engineered immune cell of any one of aspects 24 to 35, comprising a coding sequence for a cytokine, wherein the cytokine comprises IL-2, IL-7, IL- 12, IL- 15, IL- 18, IL-21, IL-23, and / or GMCSF.

[0058] Aspect 37, is the engineered immune cell of aspect 36, where the cell is modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 45), UT-NK15-28 (SEQ ID NO: 47), or UTNK15-28-DAP10 (SEQ ID NO: 49).

[0059] Aspect 38, is the engineered immune cell of aspect 37, wherein the cytokine comprises IL- 15 and / or IL-21.

[0060] Aspect 39, is the engineered immune cell of aspect 38, wherein the cytokine comprises IL-15 and comprises a polypeptide sequence and / or polynucleotide sequence encoding the same that is at least 85% identical to any one or more of SEQ ID NOs: 182-183.

[0061] Aspect 40, is the engineered immune cell of aspect 38, wherein the cytokine comprises IL-21 and comprises a polypeptide sequence and / or polynucleotide sequence encoding the same that is at least 85% identical to any one or more of SEQ ID NOs: 184-187.

[0062] Aspect 41, is the engineered immune cell of any one of aspects 24 to 40, wherein the TCR polypeptides are invariant TCR (iTCR) polypeptides.

[0063] Aspect 42, is the engineered immune cell of aspect 41, wherein the iTCRa and iTCRp polypeptides and / or polynucleotides encoding the same comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 51- 149.

[0064] Aspect 43, is the engineered immune cell of aspect 41 or 42, wherein the iTCRp polypeptide comprises a polynucleotide encoding a VP-DJ region that is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 75-149.

[0065] Aspect 44, is the engineered immune cell of any one of aspects 41 to 43, wherein the iTCRp polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NO: NOs: 59-74.

[0066] Aspect 45, is the engineered immune cell of any one of aspects 41 to 44, wherein the encoded iTCRp polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 60.

[0067] Aspect 46, is the engineered immune cell of aspect 45, wherein the polynucleotide encoding the iTCRp polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 59.

[0068] Aspect 47, is the engineered immune cell of any one of aspects 41 to 46, wherein the iTCRa polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 51-52.

[0069] Aspect 48, is the engineered immune cell of any one of aspects 24 to 47, wherein the CD16 Fc binding domain comprising polypeptide comprises a human CD16 derived Fc binding domain.

[0070] Aspect 49, is the engineered immune cell of any one of aspects 24 to 48, wherein the polypeptide comprising a CD16 derived Fc binding domain comprises a human CD16A Fc binding domain.

[0071] Aspect 50, is the engineered immune cell of any one of aspects 24 to 49, wherein the Fc binding domain is fused in N to C terminus order to an optional hinge domain, a transmembrane domain (TMD), and one or more optional intracellular signaling domains (ICD).

[0072] Aspect 51, is the engineered immune cell of aspect 50, wherein the TMD is derived from CD 16, or CD3(^.

[0073] Aspect 52, is the engineered immune cell of aspect 50 or 51, wherein the TMD comprises or consists of a sequence with at least about 90% identity to SEQ ID NOs: 163 or 167.

[0074] Aspect 53, is the engineered immune cell of any one of aspects 50 to 52, wherein the hinge domain is derived from CD32.

[0075] Aspect 54, is the engineered immune cell of any one of aspects 50 to 53, wherein the hinge domain comprises or consists of a sequence with at least about 90% identity to SEQ ID NO: 161.

[0076] Aspect 55, is the engineered immune cell of any one of aspects 50 to 54, comprising an ICD derived from CD 16 and / or CD3(^.

[0077] Aspect 56, is the engineered immune cell of any one of aspects 24 to 55 comprising a coding sequence that is least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 171-175.

[0078] Aspect 57, is the engineered immune cell of any one of aspects 24 to 56, wherein the cell is a Natural Killer (NK) cell.

[0079] Aspect 58, is the engineered NK cell of aspect 57, wherein the NK cells are derived from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or a combination thereof.

[0080] Aspect 59, is the engineered NK cell of any one of aspects 57 to 58, wherein the NK cells are primary NK cells, and are not derived from stem cells and / or induced pluripotent stem cells (iPSCs).

[0081] Aspect 60, is the engineered NK cell of any one of aspects 57 to 59, wherein the NK cells are loaded with (complexed with) one or more antibodies.

[0082] Aspect 61, is the engineered NK cell of aspect 60, wherein the one or more antibodies are one or more monospecific, bispecific, or multi-specific antibodies.

[0083] Aspect 62, is the engineered NK cell of aspect 60 or 61, wherein at least one or more antibodies comprises a glycoengineered Fc domain that has a high affinity to wild type CD 16.

[0084] Aspect 63, is the engineered NK cell of aspect 60 or 61, wherein the at least one or more antibodies comprise a non-glycoengineered Fc domain that has a low affinity to wild type CD 16 Fc binding domains.

[0085] Aspect 64, is the engineered NK cell of aspect 63, wherein the non-glycoengineered Fc domain is loaded on (complexed to) the transgenic polypeptide comprising a CD16 Fc binding domain.

[0086] Aspect 65, is the engineered NK cell of any one of aspects 60 to 64, wherein the one or more antibodies comprise an IgGl and / or IgG4 Fc domain.

[0087] Aspect 66, is the engineered NK cell of any one of aspects 60 to 65, wherein the one or more antibodies target antigens CD3, CD16, CD28, EGFR, c-MET, CD30, PSMA, MUC17, CD33, FLT3, STEAP1, BCMA, CLDN18.2, CD123, CD19, CD20, EpCAM, CEA, GPC3, CD38, CD33, CD22, HER2, GPA33, GD2, MUC16, GPRC5D, DLL-3, CLEC12A, FcRH5, and / or SSTR.

[0088] Aspect 67, is the engineered NK cell of any one of aspects 60 to 66, wherein the one or more antibodies target CD3, CD 16, CD28, CD 19, CD20, CD30, HER2, GPRC5D, EGFR, c-MET, and / or BCMA.

[0089] Aspect 68, is the engineered NK cell of any one of aspects 60 to 67, wherein the one or more antibodies comprise [fam] -trastuzumab deruxtecan, Abciximab, Adalimumab, Ado-trastuzumab emtansine, Aducanumab, Alemtuzumab, Alirocumab, Amivantamab, Anifrolumab, Ansuvimab, Atezolizumab, Atoltivimab with Maftivimab and Odesivimab-ebgn (aka Inmazeb), Avelumab, Basiliximab, Belantamab mafodotin, Belimumab, Benralizumab, Bevacizumab, Bezlotoxumab, Bimekizumab, Blinatumomab, Brentuximab vedotin, Brodalumab, Brolucizumab, Burosumab, Canakinumab, Caplacizumab, Casirivimab + imdevimab, Catumaxomab, Cemiplimab, Certolizumab pegol, Cetuximab, Cevostamab, Crizanlizumab, Daclizumab, Daratumumab, Denosumab, Dinutuximab, Donanemab, Dostarlimab, Dupilumab, Durvalumab, Eculizumab, Edrecolomab, Efalizumab, Elotuzumab, Emapalumab, Emicizumab, Enfortumab vedotin, Eptinezumab, Erenumab, Evinacumab, Evolocumab, Faricimab, Fremanezumab, Galcanezumab, Gemtuzumab, Gemtuzumab- Ozogamicin, Golimumab, Guselkumab, Ibalizumab, Ibritumomab tiuxetan, Idarucizumab, Imgatuzumab, Inebilizumab, Infliximab, Inolimomab, Inotuzumab, Inotuzumab-Ozogamicin, IPH61, Ipilimumab, Isatuximab, Ixekizumab, Lanadelumab, Lecanemab, Loncastuximab tesirine, Margetuximab, Mepolizumab, Mirvetuximab soravtansine, Mogamulizumab, Mosunetuzumab, Moxetumomab pasudotox, Murom onab-CD3, Narsoplimab, Natalizumab, Naxitamab, Nebacumab, Necitumumab, Nirsevimab, Nivolumab, Obiltoxaximab,Obinutuzumab, Ocrelizumab, Ofatumumab, Olaratumab, Omalizumab, Omburtamab, Oportuzumab monatox, Palivizumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin, Ramucirumab, Ranibizumab, Ravulizumab, Raxibacumab, Regdanvimab, Relatlimab, Reslizumab, Retifanlimab, Risankizumab, Rituximab, Romosozumab, Sacituzumab govitecan, Sarilumab, Satralizumab, Secukinumab, Siltuximab, Sintilimab, Sotrovimab, Spesolimab, Sutimlimab, Tafasitamab, Tebentafusp, Teclistamab, Teplizumab, Teprotumumab, Tezepelumab, Tildrakizumab, Tislelizumab, Tisotumab vedotin, Tixagevimab, cilgavimab, Tocilizumab, Toripalimab, Tositumomab-1131, Tralokinumab, Trastuzumab, Tremelimumab, Ublituximab, Ustekinumab, Vedolizumab, AMG 160 / Acapatamab, AMG 199 / TNB 585, AMG 330, AMG 427 / EMIRODATAMAB, AMG 509, AMG 701, AMG 910, APVO414 / ES414 / MOR209, APVO436, Catumaxomab / Removab, CC- 1, CC-93269 / EM801, Cibisatamab / RG7802 / RO6958688, CLN-049, Elranatamab / PF- 06863135, EMB-06, Epcoritamab / GEN3013, ERY974, Flotetuzumab / MGD006,Glofitamab / RG6026 / RO7082859, ISB 1342 / GBR 1342, JNJ-63709178, JNJ-63898081, JNJ- 67571244, JNJ-75348780, Linvoseltamab / REGN 5458, M701, M802, MGD007, Mosunetuzumab / RG7828, Nivatrotamab / Hu3F8-BsAb, Odronextamab / REGN1979, REGN4018, REGN5459, REGN7075, REGN5678, Talquetamab / JNJ-64407564,Tarlatamab / AMG 757, Tepoditamab / MCLA-117, TNB-383B, TNB-486, TNB-585, XmAbl3676 / Plamotamab, XmAbl4045 / Vibecotamab, XmAbl8087 / Tidutamab, and / or AFM13.

[0090] Aspect 69, is the engineered NK cell of any one of aspects 60 to 68, wherein the one or more antibodies comprise Elranatamab, Imgatuzumab, Margetuximab, Amivantamab, Blinatumomab, Obinutuzumab, IPH61 (also known as IPH6101 or SAR443579), Teclistamab, Cetuximab, Talquetamab, Pertuzumab, Trastuzumab, Tafasitamab, Brentuximab, and / or Rituximab.

[0091] Aspect 70, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Elranatamab.

[0092] Aspect 71, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Imgatuzumab.

[0093] Aspect 72, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Margetuximab.

[0094] Aspect 73, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Amivantamab.

[0095] Aspect 74, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Blinatumomab.

[0096] Aspect 75, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Obinutuzumab.

[0097] Aspect 76, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of IPH61.

[0098] Aspect 77, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Teclistamab.

[0099] Aspect 78, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Cetuximab.

[0100] Aspect 79, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Rituximab.

[0101] Aspect 80, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Talquetamab.

[0102] Aspect 81, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Pertuzumab.

[0103] Aspect 82, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Trastuzumab.

[0104] Aspect 83, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Tafasitamab.

[0105] Aspect 84, is the engineered NK cell of aspect 69, wherein the one or more antibodies comprises or consists of Brentuximab.

[0106] Aspect 85, is the engineered NK cell of any one of aspects 60 to 84, wherein the NK cell expresses the one or more antibodies.

[0107] Aspect 86, is the engineered NK cell of any one of aspects 57 to 85, wherein the NK cell is further modified to express one or more additional heterologous proteins selected from the group consisting of an antigen receptor, a cytokine, a homing receptor, a chemokine receptor, and a combination thereof.

[0108] Aspect 87, is the engineered NK cell of any one of aspects 57 to 86, wherein the NK cells are pre-activated with one or more cytokines.

[0109] Aspect 88, is the engineered NK cell of aspect 87, wherein the cytokines are IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, or a combination thereof.

[0110] Aspect 89, is the engineered NK cell any one of aspects 57 to 88, wherein the NK cell further comprises one or more engineered mutations in an endogenous gene.

[0111] Aspect 90, is the engineered NK cell of aspect 89, wherein the endogenous gene is GR, TGFBR2, CISH, and / or CD38.

[0112] Aspect 91, is a composition comprising the engineered NK cell of any one of aspects 57 to 90.

[0113] Aspect 92, is the composition aspect 91, further comprising a pharmaceutically acceptable excipient.

[0114] Aspect 93, is the composition aspect 91 or 92, wherein the composition is comprised in a delivery device.

[0115] Aspect 94, is a method of treating a disease in an individual, the method comprising the step of administering to the individual a therapeutically effective amount of any one of the cells or compositions of aspects 24 to 91.

[0116] Aspect 95, is the method of aspect 94, wherein the disease is an autoimmune disease, infection, and / or cancer.

[0117] Aspect 96, is the method of aspect 95, wherein the disease is an autoimmune disease.

[0118] Aspect 97, is the method of aspect 95.1, wherein the autoimmune disease comprises a B cell related autoimmunity, systemic lupus erythematosus (SLE), Systemic scleroderma (SSc), multiple sclerosis (MS), Grave’s disease, rheumatoid arthritis (RA), myositis, dermatomyositis, myasthenia gravis, Sjogren’s syndrome, diffuse scleroderma, inflammatory myopathy, inflammatory myopathy, ANCA-associated systemic vasculitis, antiphospholipid syndrome, immune nephritis, ITP, refractory POEMS syndrome, amyloidosis, autoimmune hemolytic anemia, and / or vasculitis.

[0119] Aspect 98, is the method of aspect 95, wherein the disease is cancer.

[0120] Aspect 99, is the method of aspect 98, wherein the cancer expresses CD 19, CD20,CD30, HER2, GPRC5D, EGFR, c-MET, and / or BCMA.

[0121] Aspect 100, is the method of aspect 98 or 99, wherein the cancer is pancreatic cancer, colorectal cancer, ovarian cancer, kidney cancer, glioblastoma, breast cancer, renal cancer, myeloma, and / or leukemia.

[0122] Aspect 101, is the method of aspect any one of aspects 95-100, further comprising administering to the individual at the same time or at different time, one or more monospecific, bispecific, and / or multispecific antibodies.

[0123] Aspect 102, is the method of aspect 101, wherein the one or more antibodies comprise Elranatamab, Imgatuzumab, Margetuximab, Amivantamab, Blinatumomab,Obinutuzumab, IPH61 (also known as IPH6101 or SAR443579), Teclistamab, Cetuximab, Talquetamab, Pertuzumab, Trastuzumab, Tafasitamab, Brentuximab, and / or Rituximab.

[0124] Aspect 103, is the method of aspect 101 or 102, wherein the one or more antibodies are administered at the same time, and / or the one or more antibodies and the engineered NK cells are complexed prior to administration to the individual.

[0125] Aspect 104, is the method of any one of aspects 101 to 103, wherein the one or more antibodies are administered more than once, including at least once at a time point after administration of the engineered NK cells.

[0126] Aspect 105, is the method of any one of aspects 101 to 104, wherein the one or more antibodies are administered more than once, including at least once at a time point before administration of the engineered NK cells.

[0127] Aspect 106, is the method of any one of aspects 101 to 105, wherein the one or more antibodies are administered more than once, including at least once at a time point before administration of the engineered NK cells and at least once at a time point after administration of the engineered NK cells.

[0128] Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTION OF THE DRAWINGS

[0129] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein. Unless otherwise noted, two-way analysis of variance (ANOVA) was employed for statistical analysis, and p-value of less than 0.05 was considered statistically significant.

[0130] FIGs. 1A-1D shows in vitro expression of, and the cytotoxic function of, invariant Natural Killer T-cell Receptors (iTCRs)-transduced NK cells. Additional experimental details are provided in Example 6 below. FIG. 1A, depicts exemplary flow cytometry plots showing expression of three different iTCR pairs, each comprising iTCRa (SEQ ID NO: 52), and either iTCRp chain 1 (SEQ ID NO: 54), iTCRp chain 2 (SEQ ID NO: 56), or iTCRp chain 3 (SEQID NO: 58), and CD3 on NK cells, using antibodies specific for iTCR target Va24 and Vpi 1 regions (y and x axis respectively). FIG. IB, depicts exemplary flow cytometry plots showing binding of Blinatumomab (y axis) to three different CD3 / iTCR complexes as described in (A), e.g., iTCRl, iTCR2, and iTCR3, respectively, expressing NK cells. NK cells were derived from three donors, cell donor 4 (CD4), cell donor 8 (CD 8), or cell donor N (CDN), respectively. The binding of Blinatumomab on CD3 was confirmed by flow cytometry using the CAR19 detection kit from Miltenyi Biotech™. FIG. 1C, depicts results of IncuCyte® live cell imaging assays that were used to measure the cytotoxicity of NK cells pre-loaded with Blinatumomab and co-transduced with iTCR and UT-NK15, against GFP-expressing Raji tumor cells at a 3: 1 effector to target ratio. T cells and iNKT cells were used as positive and negative controls, respectively. A reduction in GFP expression indicated cell death. Three NK donor lines were utilized, CB152, CB153, and CB154, respectively. NK cells were transduced with three different CD3 / iTCR complexes as described in (FIG. 1A), e.g., iTCRl, iTCR2, or iTCR3, or left non-transduced (NT) (as negative control). FIG. ID, depicts a bar chart showing residual tumor % (Y axis) for the data presented in FIG. 1C. Unpaired t test was utilized to compare iNKT with each of the different iTCRs individually. CD3 / iTCRNK cells were significantly (P = <0.01) more cytotoxic than iNKT cells (N = 3 replicates).

[0131] FIGs. 2A-2D shows the cloning of iTCR sequences from NK T Cells isolated from human cord blood, and the transgenic expression and efficacy of a subset of iTCR clones in transduced cord blood derived NK cells. Additional experimental details are provided in Example 6 below. FIG. 2A, depicts the isolation of iTCR clones from iNKT cells purified from five cord blood donors, and exemplary flow cytometry results showing expression of iTCRs on said donor iNKT cells. iNKT cells were isolated from 5 cord blood donors using iNKT isolation kit from Miltenyi Biotech™. Isolated cells were stimulated and expanded with irradiated cord blood PBMC(40 Gry) in the presence of 100 ng / ml of alpha-Galactosylceramide and 200 U / ml of IL-2. On day 7 after iNKT expansion, the purity of iNKT cultures was confirmed by iNKT specific antibodies against Va24 and Vpi l. Total mRNA was extracted from iNKT cells, and cDNAs of the VP-DJ regions were cloned and sequenced (100 individual clones). FIG. 2B, depicts sequences of iTCRp clone VP-DJ sequences. FIG. 2C, depicts the transgenic expression of Va24 and CD3 in transgenic CD3 / iTCR expressing NK cell populations from three cord blood donors. The NK cells were co-transduced with iTCRs (eight randomly selected iTCRp clones with coding sequences represented by SEQ ID NOs: 59, 61, 63, 65, 67, 69, 71, or 73, representing clones 3, 18, 24, 51, 56, 76, 93, and 96 respectively, andiTCRa coding sequence represented by SEQ ID NO: 51) and UT-NK15 on day 5. iTCR expression was confirmed by flow cytometry using iTCR specific antibodies against Va24 and antibodies against CD3. FIG. 2D, depicts a graph describing the results of IncuCyte® live cell imaging mediated measurements of cytotoxicity of NK cells co-transduced with iTCRs (as described in FIG. 2B) and UT-NK15, and pre-loaded with Blinatumomab (1 hour at 37 °C), against GFP expressing Raji cells at 3: 1 target effector ratios. T cells were used as positive control and non-transduced (NT) NK cells were utilized as negative controls. A reduction in GFP expression indicated tumor cell death.

[0132] FIGs. 3A-3C shows polynucleotide construct layouts and graphical schematics for exemplary uTNK15 and T Cell Receptor (TCR) / Fc Receptor (FcR) (TCR / FcR) vectors (e.g., vectors comprising a coding sequence for one or more TCR chains and one or more polypeptides comprising an Fc binding domain derived from an Fc Receptor). FIG. 3A, shows a vector map for multicistronic construct uTNK15-28 (SEQ ID NO: 47) comprising an open reading frame (ORF) coding region for CD3 complex proteins and IL-15. FIG. 3B, shows a vector map for multicistronic TCR / FcR constructs described herein comprising open reading frames for an alpha (a; SEQ ID NO: 51) and beta (P, clone 3; SEQ ID NO: 59) invariant T Cell Receptor (iTCR) chains and a CD16 Fc binding domain comprising polypeptide. FIG. 3C, shows a graphical schematic for uTNK15-28, and iTCR / CD16 Fc binding domain polypeptide bearing constructs: TCR / FcR #1 ( open reading frame (ORF) SEQ ID NO: 171, in vector SEQ ID NO: 177), TCR / FcR #2 (ORF SEQ ID NO: 172, in vector SEQ ID NO: 178), TCR / FcR #3 ( ORF SEQ ID NO: 173, in vector SEQ ID NO: 179), TCR / FcR #4 (ORF SEQ ID NO: 174, in vector SEQ ID NO: 180), and TCR / FcR #5 (ORF SEQ ID NO: 175, in vector SEQ ID NO: 181).

[0133] FIG. 4 shows CB-NK and T cells transduced with various TCR / FcR constructs. The top row depicts T cells expanded with anti-CD3 / 28 beads in the presence of IL-2 (50 lu / ML) that were transduced with the different TCR / FcR constructs at day 3. Transduction efficiency was determined 48 hours later by measuring the surface expression of iTCRvbl l and CD 16. The middle row depicts cord blood derived NK cells expanded with irradiated (100 Gy) UAPC feeder cells (2: 1 feeder cell : NK ratio) and recombinant human IL-2 (200 U / ml) in complete NK cell growth medium. Seven days after expansion, NK cells were transduced with the noted iTCR3 or one of TCR / FcR constructs #l-#5, comprising CD16 Fc binding domain polypeptide variant sequences SEQ ID NOs: 155-159 (encoded by SEQ ID NOs: 150-154, respectively) respectively. Transduction efficiency was determined 48 hours later by measuring surface expression of iTCRvbl l, CD3 complex, and as depicted in thebottom row, heterologous CD16 Fc binding domain comprising polypeptides (measured as anti-flag antibody against flag tagged CD 16 to avoid endogenous CD 16 background signal).

[0134] FIGs. 5A-5F shows TCR / FcR construct transduced NK cells loaded with Blinatumomab (Blina) displaying enhanced killing of CD19+ (Raji cell line) tumor cells. Cord blood derived NK cells were transduced with uTNK15, and iTCR3 (FIG. 5A) or one of TCR / FcR constructs #l-#5 (FIGs. 5B-5F). Non-transduced (NT) cord blood derived NK cells (NT NK cell) served as negative controls, while non-transduced T cells were used as positive controls. Cells were or were not loaded with Blinatumomab (100 pg / ml final concentration) for one hour at room temperature in PBS, cells were then washed prior to co-culturing with Raji cells. The Raji cells were labelled with chromium-51 and co-cultured with the immune cells at various effector to target (E:T) ratios (X axis). Cells were co-cultured for four hours and chromium release (corresponding to NK cell cytotoxicity against the cancer cells) was measured (y-axis). Compared to non-loaded TCR / FcR-NK cells, or loaded non-transduced (NT) NK Cells, Blinatumomab-loaded TCR / FcR transduced NK cells showed increased cytotoxicity against CD19+ Raji cells.

[0135] FIG. 6 shows TCR / FcR transduced NK cells loaded with Blinatumomab and / or Obinutuzumab displaying enhanced killing of CD19+++ / CD20+ (Nalm6) tumor cells. Cord blood derived NK cells were transduced with uTNK15, and iTCR3 or one of TCR / FcR constructs #l-#5. Non-transduced (NT) cord blood derived NK cells (NT NK cell) served as negative controls. NK cells were left unloaded (PBS), loaded with Blinatumomab (100 pg / ml final concentration), loaded with Obinutuzumab (500 pg / ml final concentration), or loaded with Blinatumomab (100 pg / ml final concentration) and Obinutuzumab (500 pg / ml final concentration) for one hour at room temperature in PBS and then washed prior to co-culturing with tumor cells. Immune cells and Nalm6 cells were co-cultured at a 1 : 1 E:T ratio and realtime cytotoxicity of effector cells against the Nalm6 cells was measured every 2 hours over a 40 hour period. Compared to non-loaded TCR / FcR-NK cells or loaded NT NK Cells, Blinatumomab and / or Obinutuzumab loaded TCR / FcR transduced NK cells showed increased cytotoxicity against CD 19+ NALM6 cells.

[0136] FIG. 7 shows how the T cell engager, Teclistamab, can bind to TCR / FcR transduced NK cells and T cells, but not to non-transduced NK cells. NK cells were isolated from cord blood and expanded in complete media in the presence of irradiated (100 Gy) uAPC feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL-2 (200 U / ml). Seven days following expansion, NK cells were transduced with uTNK15, and iTCR3 or one of TCR / FcR constructs #l-#5, or left non-transduced (NT). T cells were used as a positive control andloaded with Teclistamab (200 pg / ml final concentration) for one hour at room temperature in PBS and then washed prior to validation of Teclistamab binding. Flow cytometric analysis against anti-human IgG stained cells showed Teclistamab was bound to NK cells transduced with uTNK15, and iTCR3 or TCR / FcR #l-#5, and to T cells, but not NT NK cells.

[0137] FIGs. 8A-8F shows TCR / FcR transduced NK cells loaded with Teclistamab (Tecli) displaying enhanced killing of BCMA+ (MM. IS, myeloma) tumor cells. Cord blood derived NK cells were transduced with uTNK15, and iTCR3 (FIG. 8A) or one of TCR / FcR constructs #l-#5 (FIGs. 8B-8F). Non-transduced (NT) cord blood derived NK cells (NT NK cell) served as negative controls, while non-transduced T cells were used as positive controls. Cells were or were not loaded with Teclistamab (200 pg / ml final concentration) for one hour at room temperature in PBS, cells were then washed prior to co-culturing with MM. IS cells. The MM. IS cells were labelled with chromium-51 (cr51) and co-cultured with the immune cells at various effector to target (E:T) ratios (X axis). Cells were co-cultured for four hours and chromium release (corresponding to NK cell cytotoxicity against the cancer cells) was measured (y-axis). Compared to non-loaded TCR / FcR-NK cells, or loaded non-transduced (NT) NK Cells, Teclistamab-loaded TCR / FcR transduced NK cells showed increased cytotoxicity against BCMA+ MM.1 S cells.

[0138] FIGs. 9A-9F shows TCR / FcR transduced NK cells loaded with Teclistamab (Tecli) displaying enhanced killing of BCMA+ (H929, myeloma) tumor cells. Cord blood derived NK cells were transduced with uTNK15, and iTCR3 (FIG. 9A) or one of TCR / FcR constructs #1- #5 (FIGs. 9B-9F). Non-transduced (NT) cord blood derived NK cells (NT NK cell) served as negative controls, while non-transduced T cells were used as positive controls. Cells were or were not loaded with Teclistamab (200 pg / ml final concentration) for one hour at room temperature in PBS, cells were then washed prior to co-culturing with MM.1 S cells. The H929 cells were labelled with chromium-51 and co-cultured with the immune cells at various effector to target (E:T) ratios (X axis). Cells were co-cultured for four hours and chromium release (corresponding to NK cell cytotoxicity against the cancer cells) was measured (y-axis). Compared to non-loaded TCR / FcR-NK cells, or loaded non-transduced (NT) NK Cells, Teclistamab-loaded TCR / FcR transduced NK cells showed increased cytotoxicity against BCMA+ H929 cells.

[0139] FIG. 10, shows binding of low Fey receptor affinity antibody (e.g., Cetuximab) to TCR / FcR transduced NK cells. NK cells were derived from cord blood and expanded with irradiated (100 Gy) UAPC feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL- 2 (200 U / ml) in complete NK cell growth medium (Click’ s / RPMI). NK cells were loaded withlow Fey receptor affinity antibody (e.g., Cetuximab at 100 pg / ml final concentration) for one hour at 37 °C in Click’ s / RPMI media followed by washing before validating the binding of the low Fey receptor affinity antibody with flow cytometry. NK cells were stained with Alexa- Fluor647 affinity-purified F(ab’)2 fragment goat anti-human IgG (H+L) antibody and analyzed by flow cytometry. The results showed that antibodies with low Fey receptor affinity Fc domains (e.g., Cetuximab) bound to high affinity CD16 (CD16ha, (F158V)) TCR / FcR transduced NK cells at greater levels when compared to ITCR3 transduced NK cells, NT NK cells, or T cells.

[0140] FIG. 11, shows TCR / FcR transduced NK cells loaded with Cetuximab displaying enhanced killing of WiDR (colorectal cancer cells; CRC) compared to non-loaded TCR / FcR NK cells or loaded / non-loaded non-transduced (NT) NK Cells. NK cells were transduced with uTNK15, and iTCR3 or one of TCR / FcR constructs #l-#5 . WiDR cells alone, Non-transduced (NT) cord blood derived NK cells (NT NK cell), and T cells served as served as controls. Cells were or were not loaded with Cetuximab (100 pg / ml final concentration) for one hour at 37 °C in Click’ s / RPMI media and then washed before co-culturing with tumor cells at 1 : 1 E:T ratio. Compared to Cetuximab alone, or to loaded / non-loaded NT NK cells or to non-loaded TCR / FcR NK cells, Cetuximab loaded TCR / FcR NK cells showed the highest cytotoxic activities against the EGFR+ CRC (WiDR) cell line. The lower the normalized cell index (Y axis), the higher the degree of cytotoxicity. SDS was used a positive control for cytotoxicity. NK cells transduced with TCR / FcR #4 or TCR / FcR #2 and loaded with cetuximab exerted the greatest levels of cytotoxicity against WiDR cells.

[0141] FIGs. 12A-12B, shows TCR / FcR transduced NK cells loaded with Cetuximab displaying enhanced killing of PATC148 cells (pancreatic ductal adenocarcinoma, PDAC) compared to non-loaded TCR / FcR NK cells or loaded / non-loaded non-transduced (NT) NK Cells. NK cells were transduced with uTNK15, and iTCR3 or one of TCR / FcR constructs #1- #5. PATC148 cells alone, Non-transduced (NT) cord blood derived NK cells (NT NK cell), and T cells served as controls. Cells were or were not loaded with Cetuximab (100 pg / ml final concentration) for one hour at 37 °C in Click’ s / RPMI media and then washed before coculturing with tumor cells at 4: 1 E:T ratio (FIG. 12A), or at 2: 1 ET ratio (FIG. 12B). Compared to Cetuximab alone, or with loaded / non-loaded non-transduced (NT) NK cells or with nonloaded TCR / FcR NK cells, Cetuximab loaded TCR / FcR transduced NK cells showed the greatest cytotoxic activity against EGFR+ PATC148 (PDAC) cell lines. The lower the normalized cell index (Y axis), the higher the degree of cytotoxicity. SDS was used a positivecontrol for cytotoxicity. NK cells transduced with TCR / FcR #4 or TCR / FcR #2 and loaded with cetuximab exerted the greatest levels of cytotoxicity against PATC148 (PDAC) cells.

[0142] FIGs. 13A-13B show TCR / FcR transduced NK cells loaded with Imgatuzumab displaying enhanced killing of PDAC or CRC cells when compared to non-loaded TCR / FcR NK cells or loaded / un-loaded non-transduced (NT) NK Cells. NT NK cells or NK cells transduced with uTNK 15 and TCR / FcR construct #2 were loaded with Imgatuzumab (10 pg / ml final concentration) for one hour at room temperature in PBS and then washed before co-culture with EGFR+ PDAC (PATC-148, FIG. 13A) or EGFR+ colorectal cancer (WiDR, FIG. 13B) tumor cells at a 1 : 1 E:T ratio. Compared to Imgatuzumab alone, loaded / un-loaded NT-NK cells, or non-loaded TCR / FcR NK cells, Imgatuzumab loaded TCR / FcR transduced NK cells showed increased cytotoxic activity against EGFR+ PDAC (PATC148) and colorectal (WiDR) cell lines.

[0143] FIGs. 14A-14C show TCR / FcR transduced NK cells loaded with Imgatuzumab displaying enhanced killing of 3D PDAC (PATC-148) tumor spheroids. FIG. 14A, displays representative images of PATC148 spheroids (PDAC tumor cell line transduced with GFP) either left alone, treated with NT NK Cells, or treated with NK cells transduced with uTNK15 and TCR / FcR construct #2, the NK cells were either non-loaded or loaded with Imgatuzumab (10 pg / ml final concentration) for one hour at room temperature in PBS and then washed before co-culture. The data showed significant decreases in spheroid size in the wells treated with Imgatuzumab loaded TCR / FcR transduced NK cells. FIG. 14B, depicts quantification of the total integrated green intensity over time observed in 14A, the data demonstrated a significant decrease in total integrated green intensity (FIG. 14C) when spheroids were treated with TCR / FcR transduced NK cells loaded with Imgatuzumab.

[0144] FIGs. 15A-15C, shows binding of Cetuximab, Amivantamab, or Imgatuzumab to TCR / FcR #2 transduced NK cells. NK cells were derived from cord blood and were expanded with irradiated (100 Gy) UAPC feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL-2 (200 U / ml) in complete NK cell growth medium (Click’ s / RPMI). Non-transduced NK cells or NK cells transduced with TCR / FcR #2 were loaded with Cetuximab (250 pg / ml final concentration) (FIG. 15A), Amivantamab (100 pg / ml final concentration) (FIG. 15B), or Imgatuzumab (100 pg / ml final concentration) (FIG. 15C), for one hour at 37 °C in Click’ s / RPMI media before the cells were washed. NK cells were stained with Alexa-Fluor647 affinity-purified F(ab’)2 fragment goat anti-human IgG (H+L) antibody and analyzed by flow cytometry to confirm the binding of Cetuximab, Amivantamab, or Imgatuzumab. The results showed that antibodies were bound to TCR / FcR transduced NK cells with higher affinity whencompared to NT NK cells. Of note, unlike Amivantamab or Imgatuzumab, Cetuximab (which comprises an Fc domain with low binding affinity to WT CD 16) was found to bind only to the CD16ha TCR / FcR transduced NK cells.

[0145] FIGs. 16A-16C, depicts uTNK15 and TCR / FcR #2 transduced NK cells demonstrating enhanced antitumor activity against PATC-148 (PDAC) cell spheroids when loaded with Cetuximab (Cetux), Imgatuzumab (Imga), or Amivantamab (Ami) relative to NT NK cells. Antibody loaded TCR / FcR transduced NK cells showed enhanced killing of PDAC cells when compared to non-loaded TCR / FcR NK cells or loaded / non-loaded non-transduced (NT) NK Cells. FIG. 16A shows representative images of GFP transduced PDAC (PATC-148) spheroids that were either left alone, treated with non-loaded or loaded NT NK, or treated with non-loaded or loaded TCR / FcR #2 transduced NK cells. Loaded cells were prepared with Imgatuzumab (100 pg / ml final concentration), Amivantamab (100 pg / ml final concentration), or Cetuximab (250 pg / ml final concentration). A readily observable and significant decrease in spheroid size was observed in the wells treated with antibody-loaded TCR / FcR transduced NK cells FIG. 16B is a graphical quantification of the total integrated green intensity over time from experiments as performed in FIG. 16A, the data shows a significant decrease (FIG. 16C) in total integrated green intensity when spheroids were treated with TCR / FcR transduced NK cells loaded with antibodies.

[0146] FIGs. 17A-17C depicts uTNK15 and TCR / FcR #2 transduced NK cells demonstrating enhanced antitumor activity against WiDR (CRC) cell spheroids when loaded with Cetuximab (Cetux), Imgatuzumab (Imga), or Amivantamab (Ami) relative to NT NK cells. Antibody loaded TCR / FcR transduced NK cells showed enhanced killing of CRC cells when compared to non-loaded TCR / FcR NK cells or loaded / non-loaded non-transduced (NT) NK Cells. FIG. 17A shows representative images of GFP transduced WiDR spheroids that were either left alone, treated with non-loaded or loaded NT NK, or treated with non-loaded or loaded TCR / FcR #2 transduced NK cells. Loaded cells were prepared with Imgatuzumab (100 pg / ml final concentration), Amivantamab (100 pg / ml final concentration), or Cetuximab (250 pg / ml final concentration). The data show significant decreases in spheroid size in the wells treated with antibody-loaded TCR / FcR transduced NK cells. FIG. 17B is a graphical quantification of the total integrated green intensity over time from experiments as performed in FIG 16A, the data shows a significant decrease (FIG. 17C) in total integrated green intensity when spheroids were treated with TCR / FcR transduced NK cells loaded with antibodies.

[0147] FIGs. 18A-18F, depicts uTNK15 and TCR / FcR #2 transduced NK cells demonstrating enhanced antitumor activity against various solid tumor cell lines when loadedwith anti-EGFR antibodies Cetuximab, Imgatuzumab, or Amivantamab. Antibody loaded TCR / FcR transduced NK cells showed enhanced killing when compared to non-loaded TCR / FcR NK cells or loaded / non-loaded non-transduced (NT) NK Cells. NK cells were either non-transduced (NT) or transduced with TCR / FcR #2 and were left unloaded or loaded with Cetuximab (250 pg / ml final concentration), Amivantamab (100 pg / ml final concentration), or Imgatuzumab (100 pg / ml final concentration) for one hour at 37 °C in Click’ s / RPMI media followed by washing. NK cells were co-cultured with tumor cells SKOV3 (FIG. 18A), PATC- 148 (FIG. 18C), or WiDR (FIG. 18E) at 2: 1 E:T ratios. Compared to antibodies alone, loaded / non-loaded non-transduced (NT) NK cells, or non-loaded TCR / FcR #2 NK cells, the antibody-loaded TCR / FcR #2 transduced NK cells showed significantly increased cytotoxic activity against EGFR+ SKOV3 (FIG. 18B), PATC-148 (FIG. 18D), or WiDR (FIG. 18F) cell lines.

[0148] FIGs. 19A-19B, shows binding of low Fey receptor affinity antibody (e.g., Rituximab) or high Fey receptor affinity antibody (Obinutuzumab) to TCR / FcR #2 transduced NK Cells. NK cells were derived from cord blood and were expanded with irradiated (100 Gy) UAPC feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL-2 (200 U / ml) in complete NK cell growth medium (Click’ s / RPMI). Non-transduced NK cells or NK cells transduced with TCR / FcR #2 were loaded with Rituximab (100 pg / ml final concentration) (FIG. 19A), or Obinutuzumab (100 pg / ml final concentration) (FIG. 19B) for one hour at 37 °C in Click’ s / RPMI media followed by washing. NK cells were stained with Alexa-Fluor647 affinity-purified F(ab’)2 fragment goat anti-human IgG (H+L) antibody and analyzed by flow cytometry. The results showed that low and high Fey receptor affinity antibodies were bound to TCR / FcR transduced NK cells at greater rates when compared to NT NK cells. Of note, unlike Obinutuzumab, Rituximab (which comprises an Fc domain with low binding affinity to WT CD16) was found to bind only to the CD16ha TCR / FcR transduced NK cells.

[0149] FIGs. 20A-20B, depicts TCR / FcR #2 transduced NK cells demonstrating enhanced antitumor activity against Raji (B cell lymphoma) cells when loaded with anti-CD20 antibodies Rituximab or Obinutuzumab when compared to NT NK at various E:T ratios. NK cells were either non-transduced (NT) or transduced with TCR / FcR #2, and left unloaded or loaded with Rituximab (100 pg / ml final concentration) (FIG. 20 A), or Obinutuzumab (100 pg / ml) for one hour at 37 °C in Click / RPMI media followed by washing. NK cells were co-cultured with chromium-51 labelled Raji cells at various E:T ratios (5: 1, 2: 1, 1 : 1, or 1 :2), for four hours and chromium release (corresponding to the cytotoxicity of the cancer cells) was measured.Compared to non-loaded TCR / FcR NK cells or loaded NT NK cells, antibody-loaded TCR / FcR #2 transduced NK cells showed increased cytotoxicity against CD20+ Raji cells.

[0150] FIG. 21, depicts an exemplary combination of therapeutic strategies as described herein (e.g., uTNK15 and TCR / FcR transduced NK cells loaded with antibodies).

[0151] FIGs. 22A-22B, shows uTNK15 and TCR / FcR transduced NK cells loaded with Elranatamab demonstrated enhanced antitumor activity against BCMA+ multiple myeloma cells (e.g., MM. IS cells), and outlines a clinical trial comprising combination TCR / FcR transduced NK cells and Elranatamab. FIG. 22A shows the results of a Bioluminescence imaging (BLI) experiment, where mice were irradiated, inoculated with 5 x 105MM. IS cells and sham injected or injected (IV) with a single infusion of 1 x 107NK cells that were either non-transduced or transduced with uTNK15 and TCR / FcR #2. Animals were imaged weekly. The results showed that mice receiving TCR / FcR transduced NK cells loaded with Elranatamab demonstrated enhanced antitumor activity in vivo relative to controls. FIG. 22B schematically outlines an exemplary clinical trial of engineered uTNK15 and TCR / FcR transduced NK cells (““TCR / FcR #2 NK cell” shorthand) coupled with Elranatamab.

[0152] FIG. 23, depicts binding of T cell engagers (e.g., Teclistamab (anti-BCMA), Elranatamab (anti-BCMA), and Blinatumumab (anti-CD19)) to uTNK15 and TCR / FcR #2 transduced NK cells but not to NT NK cells. Natural killer (NK) cells were extracted from cord blood and cultured in complete medium supplemented with irradiated (100 Gy) uAPC feeder cells at a ratio of 2: 1 (feeder cell:NK). Additionally, recombinant human IL-2 was added at a concentration of 200 U / ml to facilitate expansion of the NK cells. Five days after expansion, NK cells were either not transduced, or transduced with the uTNK15 and TCR / FcR #2. These cells were then treated with Teclistamab (20 pg / ml), Elranatamab (10 pg / ml), or Blinatumumab (10 pg / ml) for one hour at 37 °C in a complete medium. After incubation, the cells were washed and subjected to flow cytometric analysis to validate the binding. Before running on flow cytometry, cells that had been incubated with BCMA or CD 19 antigen with His tag were stained with APC anti-His tag antibody (cat no: 362605, Biolegend). Flow cytometric analysis revealed that all T cell engagers could bind efficiently to engineered NK cells, but no binding was observed with NT NK cells.

[0153] FIGs. 24A-24B, depict uTNKl 5 and TCR / FcR #2 transduced NK cells loaded with bispecific anti-CD19 / CD3 antibody blinatumomab (10 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) demonstrating enhanced antitumor activity against CD 19+ tumor cells (e.g., Raji cells or B-LCL cells) at various E:T ratios in short term Cr51 assays when compared to blinatumomab loaded NT NK cells. FIG. 24A shows the results against B-LCL cells, where there was significantly more killing by engineered NK cells at 5: 1, 2.5: 1, and 1.25: 1 E:T ratios. FIG. 24B shows the results against Raji cells, where there was significantly more killing by engineered NK cells at 5: 1 and 2.5: 1 E:T ratios. Compared to blinatumomab- loaded NT NK cells, blinatumomab-loaded engineered NK cells showed enhanced cytotoxicity against CD 19+ tumor cells.

[0154] FIG. 25, Blinatumomab loaded uTNK15 and TCR / FcR #2 transduced NK cells showed enhanced killing compared to unloaded uTNKl 5 and TCR / FcR #2 transduced NK cells or loaded / unloaded non-transduced (NT) NK Cells. NK cells were either NT (NT or NT NK shorthand for stats) or transduced with uTNKl 5 and TCR / FcR #2 (“TCR / FcR #2” shorthand for stats). NK cells were either left unloaded or loaded with blinatumomab (10 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) before co-culturing with tumor cells at 1 : 1 effector to target (E:T) ratio. Compared to loaded or unloaded NT NK Cells, unloaded engineered NK cells, or Blinatumumab alone, Blinatumumab-loaded engineered NK cells showed significantly increased cytotoxic activity against CD 19 transduced SKOV3 cancer cells.

[0155] FIGs. 26A-26B, depict uTNKl 5 and TCR / FcR #2 transduced NK cells loaded with bispecific anti-BCMA / CD3 antibody Teclistamab (20 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) demonstrating enhanced antitumor activity against BCMA+ tumor cells (e.g., MMls cells, H929 cells) at various E:T ratios in short term Cr51 assays when compared to Teclistamab loaded non-transduced (NT) NK cells. FIG. 26A shows the results against MMls cells, where there was significantly more killing by engineered NK cells at 5: 1, 2.5: 1, and 1.25: 1 E:T ratios. FIG. 26B shows the results against H929 cells, where there was significantly more killing by engineered NK cells at 5: 1, 2.5: 1, 1.25: 1, and 1 :2 E:T ratios. Compared to Teclistamab-loaded NT NK cells, Teclistamab-loaded engineered NK cells showed enhanced cytotoxicity against BCMA+ tumor cells.

[0156] FIGs. 27A-27B, depict uTNKl 5 and TCR / FcR #2 transduced NK cells loaded with bispecific anti-GPRC5D / CD3 antibody Talquetamab (20 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) demonstrating enhanced antitumor activity against GPRC5D+ tumor cells (e.g., MMls cells, H929 cells) at various E:T ratios in a short term Cr51 assays when compared to Talquetamab loaded non-transduced (NT) NK cells. FIG. 27A shows the results against MMls cells. FIG. 27B shows the results against H929 cells. Compared to Talquetamab-loaded NT NK cells, Talquetamab-loaded engineered NK cells showed enhanced cytotoxicity against GPRC5D+ tumor cells.

[0157] FIGs. 28A-28B, depict uTNKl 5 and TCR / FcR #2 transduced NK cells loaded with bispecific anti-BCMA / CD3 antibody Elranatamab (10 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) demonstrating enhanced antitumor activity against BCMA+ tumor cells (e.g., MMls cells, H929 cells) at various E:T ratios in short term Cr51 assays when compared to Elranatamab loaded non-transduced (NT) NK cells. FIG. 28A shows the results against MMls cells, where there was significantly more killing by engineered NK cells at 5:1, 2.5: 1, 1.25: 1, and 1 :2 E:T ratios. FIG. 28B shows the results against H929 cells, where there was significantly more killing by engineered NK cells at 5: 1, 2.5: 1, 1.25: 1, and 1 :2 E:T ratios. Compared to Elranatamab-loaded NT NK cells, Elranatamab-loaded engineered NK cells showed enhanced cytotoxicity against BCMA+ tumor cells.

[0158] FIG. 29A-29B, Pertuzumab (anti-HER2 Ab with low affinity binding to wild type CD16) bound to uTNK15 and TCR / FcR #2 transduced NK cells, but not to non-transduced (NT) NK cells, and Pertuzumab loaded engineered NK cells displayed superior antitumor activity against HER2+ SKOV3 ovarian cancer cells in a long term xCELLigence killing assay. FIG. 29 A, shows Pertuzumab binding to engineered NK cells. NK cells were extracted from cord blood and cultured in complete medium supplemented with irradiated (100 Gy) uAPC feeder cells at a ratio of 2: 1 (feeder cell:NK). Additionally, recombinant human IL-2 was added at a concentration of 200 U / ml to facilitate expansion of the NK cells. Five days after expansion, natural killer (NK) cells were either transduced with the uTNKl 5 and TCR / FcR #2 constructs, or left non-transduced (NT). These cells were then loaded with Pertuzumab (200 pg / ml for one hour at 37 °C in a complete medium). After incubation, the cells were washed and subjected to flow cytometric analysis to validate the binding. Flow cytometric analysis revealed that Pertuzumab could bind engineered NK cells, but no binding was observed with NT NK cells. FIG. 29B, Pertuzumab loaded uTNKl 5 and TCR / FcR #2 transduced NK cells showed enhanced killing compared to unloaded uTNKl 5 and TCR / FcR #2 transduced NK cells or loaded / unloaded non-transduced (NT) NK Cells. NK cells were either NT (NT or NT NK shorthand for stats) or transduced with uTNKl 5 and TCR / FcR #2 (“TCR / FcR #2” shorthand for stats). NK cells were either left unloaded or loaded with Pertuzumab (200 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) before co-culturing with tumor cells at 1 : 1 effector to target (E:T) ratio. Compared to loaded or unloaded NT NK Cells, unloaded engineered NK cells, or Pertuzumab alone, Pertuzumab-loaded engineered NK cells showed significantly increased cytotoxic activity against HER2+ SKOV3 ovarian cancer cells.

[0159] FIGs. 30A-30B, Trastuzumab (anti-HER2) bound uTNKl 5 and TCR / FcR #2 transduced NK cells with higher affinity than non-transduced (NT) NK cells, and Trastuzumabloaded engineered NK cells displayed superior antitumor activity against HER2+ SKOV3 ovarian cancer cells in a long term xCELLigence killing assay. FIG. 30A, shows Trastuzumab binding to engineered NK cells. NK cells were extracted from cord blood and cultured in complete medium supplemented with irradiated (100 Gy) uAPC feeder cells at a ratio of 2: 1 (feeder cell:NK). Additionally, recombinant human IL-2 was added at a concentration of 200 U / ml to facilitate expansion of the NK cells. Five days after expansion, natural killer (NK) cells were either transduced with the uTNK15 and TCR / FcR #2 constructs, or left non-transduced (NT). These cells were then loaded with Trastuzumab (200 pg / ml for one hour at 37 °C in a complete medium). After incubation, the cells were washed and subjected to flow cytometric analysis to validate the binding. Flow cytometric analysis revealed that Trastuzumab could bind engineered NK cells with higher affinity than was observed with NT NK cells. FIG. 30B, Trastuzumab loaded uTNK15 and TCR / FcR #2 transduced NK cells showed enhanced killing compared to unloaded uTNK15 and TCR / FcR #2 transduced NK cells or loaded / unloaded NT NK Cells. NK cells were either NT (“NT” or “NT NK” shorthand for stats) or transduced with uTNK15 and TCR / FcR #2 (“TCR / FcR #2” shorthand for stats). NK cells were either left unloaded or loaded with Trastuzumab (200 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) before co-culturing with tumor cells at 1 : 1 effector to target (E:T) ratio. Compared to loaded or unloaded NT NK Cells, unloaded engineered NK cells, or Trastuzumab alone, Trastuzumab-loaded engineered NK cells showed significantly increased cytotoxic activity against HER2+ SKOV3 ovarian cancer cells.

[0160] FIGs. 31A-31B, Tafasitamab (anti-CD19) bound uTNK15 and TCR / FcR #2 transduced NK cells with higher affinity than non-transduced (NT) NK cells, and Tafasitamab loaded engineered NK cells displayed superior antitumor activity against transduced CD 19+ SKOV3 ovarian cancer cells in a long term xCELLigence killing assay. FIG. 31A, NK cells were extracted from cord blood and cultured in complete medium supplemented with irradiated (100 Gy) uAPC feeder cells at a ratio of 2: 1 (feeder cell:NK). Additionally, recombinant human IL-2 was added at a concentration of 200 U / ml to facilitate expansion of the NK cells. Five days after expansion, natural killer (NK) cells were either left non-transduced, or transduced with the uTNK15 and TCR / FcR #2 constructs. These cells were then treated with Tafasitamab (10 pg / ml for 1 hour at 37 °C in complete media). After incubation, the cells were washed and subjected to flow cytometric analysis to validate the binding. Flow cytometric analysis revealed that Tafasitamab exhibited higher and stronger binding affinity towards engineered NK cells compared to NT NK cells. FIG. 31B, Tafasitamab loaded uTNK15 and TCR / FcR #2 transduced NK cells showed enhanced killing compared to unloaded uTNK15 and TCR / FcR#2 transduced NK cells or loaded / unloaded NT NK Cells. NK cells were either NT (“NT” or “NT NK” shorthand for stats) or transduced with uTNK15 and TCR / FcR #2 (“TCR / FcR #2” shorthand for stats). NK cells were either left unloaded or loaded with Tafasitamab (200 pg / ml for 1 hour at 37 °C in click / RPMI media & washed prior to co-culture) before co-culturing with tumor cells at 1 : 1 effector to target (E:T) ratio. Compared to loaded or unloaded NT NK Cells, unloaded engineered NK cells, or Tafasitamab alone, Tafasitamab-loaded engineered NK cells showed significantly increased cytotoxic activity against transduced CD 19+ SKOV3 ovarian cancer cells.

[0161] FIGs. 32A-32B, Brentuximab (anti-CD30 antibody-drug conjugate) bound to uTNK15 and TCR / FcR #2 transduced NK cells with higher affinity than non-transduced (NT) NK cells, and Brentuximab loaded engineered NK cells displayed superior antitumor activity against CD30+ Karpas tumor cells in IncuCyte cytotoxicity assays. FIG. 32A, NK cells were isolated from cord blood and expand in complete media in the presence of irradiated (100 Gy) uAPC feeder cells (2: 1 feeder cell:NK ratio) and recombinant human IL-2 (200 U / ml). Seven days following expansion, NK cells were either left non-transduced, or transduced with the uTNK15 and TCR / FcR #2 constructs. These cells were loaded with Brentuximab (100 pg / ml for one hour at 37 °C in complete medium) and washed prior to flow cytometric validation of the Brentuximab binding. Brentuximab bound to engineered NK cells with higher affinity compared to NT NK cells, as determined by staining with F(ab’)2 Anti-Human antibody and flow cytometric analysis of NK cells. FIG. 32B, Brentuximab-loaded uTNK15 and TCR / FcR #2 transduced NK cells showed enhanced killing of CD30+ (Karpas) tumor cells compared to non-loaded engineered NK cells or brentuximab-loaded NT NK Cells in IncuCyte cytotoxicity assays. NK cells were derived from cord blood expanded as described in 31 A, and either not transduced or transduced with uTNK15 and TCR / FcR #2 constructs. NT NK cells or engineered NK cells were loaded with Brentuximab (100 pg / ml for one hour at 37 °C in complete medium and washed prior to co-culturing) and co-cultured with CD30+ Karpas tumor cell line at 1 : 1 effector to target ratios. Real-time cytotoxicity of effector cells against Karpas cells was measured every 2 hour over 24-hour period. Compared to non-loaded engineered NK cells or NT NK cells loaded with Brentuximab, engineered NK cells loaded with Brentuximab showed increased cytotoxicity of CD30+ Karpas cells.DETAILED DESCRIPTION

[0162] In keeping with long-standing patent law convention, the words “a” and “an” when used in the present specification in concert with the word comprising, including the claims,denote “one or more.” Some embodiments of the disclosure may consist of or consist essentially of one or more elements, method steps, and / or methods of the disclosure. It is contemplated that any method or composition described herein can be implemented with respect to any other method or composition described herein and that different embodiments may be combined.

[0163] Throughout this specification, unless the context requires otherwise, the words “comprise”, “comprises” and “comprising” will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements. By “consisting of’ is meant including, and limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of’ indicates that the listed elements are required or mandatory, and that no other elements may be present. By “consisting essentially of’ is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase “consisting essentially of’ indicates that the listed elements are required or mandatory, but that no other elements are optional and may or may not be present depending upon whether or not they affect the activity or action of the listed elements.

[0164] Reference throughout this specification to “one embodiment,” “an embodiment,” “a particular embodiment,” “a related embodiment,” “a certain embodiment,” “an additional embodiment,” or “a further embodiment” or combinations thereof means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the appearances of the foregoing phrases in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0165] As used herein, the terms “or” and “and / or” are utilized to describe multiple components in combination or exclusive of one another. For example, “x, y, and / or z” can refer to “x” alone, “y” alone, “z” alone, “x, y, and z,” “(x and y) or z,” “x or (y and z),” or “x or y or z.” It is specifically contemplated that x, y, or z may be specifically excluded from an embodiment.

[0166] Throughout this application, the term “about” is used according to its plain and ordinary meaning in the area of cell and molecular biology to indicate that a value includes the standard deviation of error for the device or method being employed to determine the value.

[0167] As used herein, the term “CD3 receptor complex” or “CD3 co-receptor complex” refers to the protein complex that in nature acts as a T cell co-receptor and is comprised of CD3^ chain, CD3y chain, a CD35 chain, and two CD3s chains (although in alternatives only one CD3s chain is used).

[0168] The term “engineered” as used herein refers to an entity that is generated by the hand of man, including a cell, nucleic acid, polypeptide, vector, and so forth. In at least some cases, an engineered entity is synthetic and comprises elements that are not naturally present or configured in the manner in which it is utilized in the disclosure. In specific embodiments, a vector is engineered through recombinant nucleic acid technologies, and a cell is engineered through transfection or transduction of an engineered vector. Cells may be engineered to express heterologous proteins that are not naturally expressed by the cells, either because the heterologous proteins are recombinant or synthetic or because the cells do not naturally express the proteins.

[0169] The phrases “pharmaceutical or pharmacologically acceptable” refers to molecular entities and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal, such as a human, as appropriate. The preparation of a pharmaceutical composition comprising an antibody or additional active ingredient will be known to those of skill in the art in light of the present disclosure. Moreover, for animal (e.g., human) administration, it will be understood that preparations should meet sterility, pyrogenicity, general safety, and purity standards as required by FDA Office of Biological Standards.

[0170] As used herein, “pharmaceutically acceptable carrier” includes any and all aqueous solvents (e.g., water, alcoholic / aqueous solutions, saline solutions, parenteral vehicles, such as sodium chloride, Ringer’s dextrose, etc.), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oil, and injectable organic esters, such as ethyloleate), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, anti-oxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, fluid and nutrient replenishers, such like materials and combinations thereof, as would be known to one of ordinary skill in the art. The pH and exact concentration of the various components in a pharmaceutical composition are adjusted according to well-known parameters.

[0171] The term “subject,” as used herein, generally refers to an individual having or is suspected of having cancer. The subject can be any organism or animal subject that is an object of a method or material, including mammals, e.g., humans, laboratory animals (e.g., primates, rats, mice, rabbits), livestock (e.g., cows, sheep, goats, pigs, turkeys, and chickens), household pets (e.g., dogs, cats, and rodents), horses, and transgenic non-human animals. The subject can be a patient, e.g., have or be suspected of having a disease (that may be referred to as a medical condition), such as benign or malignant neoplasias, or cancer. The subject may be undergoing or having undergone treatment. The subject may be asymptomatic. The subject may be healthy individuals but that are desirous of prevention of cancer. The term “individual” may be used interchangeably, in at least some cases. The “subject” or “individual”, as used herein, may or may not be housed in a medical facility and may be treated as an outpatient of a medical facility. The individual may be receiving one or more medical compositions via the internet. An individual may comprise any age of a human or non-human animal and therefore includes both adult and juveniles (i.e., children) and infants and includes in utero individuals. It is not intended that the term connote a need for medical treatment, therefore, an individual may voluntarily or involuntarily be part of experimentation whether clinical or in support of basic science studies.

[0172] As used herein “treatment” or “treating,” includes any beneficial or desirable effect on the symptoms or pathology of a disease or pathological condition, and may include even minimal reductions in one or more measurable markers of the disease or condition being treated, e.g., cancer. Treatment can involve optionally either the reduction or amelioration of one or more symptoms of the disease or condition, or the delaying of the progression of the disease or condition. “Treatment” does not necessarily indicate complete eradication or cure of the disease or condition, or associated symptoms thereof. Treating may mean alleviation of at least one symptom of the disease or condition.

[0173] As used herein “TCR / CD3 complex” refers to a protein complex naturally found on the surface of T cells and that comprises T-cell receptor (TCR) a and 0 chains, invariant Natural Killer T-cell receptor (iTCR) a and 0 chains, and / or a T-cell receptor y and 5 chains, in addition to CD3^, CD3y, CD36, and CD3s chains.I. Embodiments of the Disclosure

[0174] Natural killer (NK) cells are an emerging cellular immunotherapy for patients with malignant hematologic disease, as well as solid tumors. The present disclosure specifically relates to NK cells that have been modified to render the NK cells to have enhanced functionas an immunotherapy compared to NK cells not so modified. The modifications allow for the NK cells to have greater versatility when used with other therapeutic agents and at least in some embodiments to have T cell-like activity by utilizing the CD3 / TCR receptor complex. In specific embodiments, the NK cells are modified to express (i) either a single CD3 chain (CD3zeta, CD3 epsilon, CD3 delta, or CD3 gamma) or part or all of the human CD3 receptor complex (including any combination of CD3 delta, epsilon (one or two copies of epsilon), gamma, and zeta); or (ii) either a single CD3 chain or the human CD3 receptor complex (including any combination of CD3 delta, epsilon (one or two molecules), gamma, and zeta) as a full length protein or as a partial protein heterologously linked to one or more intracellular signaling domains); and (iii) the CD3 complex may or may not include the T-cell receptor (aP or y5) and / or iTCR receptor (a|3). The disclosure concerns the use of CD3 -expressing NK cells in the diagnosis and treatment of disease, including use of the cells in combination with bispecific or multi-specific antibodies in which one epitope of the antibody binds CD3 on the CD3 -expressing NK cells. The CD3 -expressing NK cells can either be pre-complexed ex vivo with the bi / multi-specific antibody to redirect their specificity toward the target antigen and / or combined in vivo. In diagnostic embodiments, labeled NK cells may be loaded with bispecific or multi-specific antibodies of any kind, including that comprise at least an anti-CD3 antibody, and the loaded, labeled NK cells may be monitored for trafficking to the site of the target antigen for which another antibody on the bispecific or multi-specific antibody binds.

[0175] In certain embodiments of the disclosure, a TCR recognizes antigens and / or epitopes presented by a major-histocompatibility complex (MHC). In certain embodiments, an antigen and / or epitope is a peptide, lipid, and / or glycolipid. In certain embodiments, a MHC is a class I MHC. In certain embodiments, a MHC is a class II MHC. In certain embodiments, an MHC is a non-classical MHC. In certain embodiments, an MHC is a class I-like MHC. In certain embodiments, an MHC is CD Id.

[0176] In certain embodiments, a TCR target antigen is not primarily what provides a transduced effector cell with target antigen specificity. In certain embodiments, a TCR acts primarily as a stabilizer for a CD3 co-receptor complex, while an antibody provides the primary target antigen specificity for a transduced effector cell.IL Compositions of the Disclosure

[0177] The disclosure concerns compositions that at least include modified NK cells that express at least parts of the TCR / CD3 complex. In some cases, the compositions also include monospecific, bispecific, and / or multi-specific antibodies, including in the same formulation,although in alternative embodiments the NK cells and antibodies are utilized as physically separate compositions.A. NK Cell TCR / CD3 Modifications

[0178] In particular embodiments, provided herein are NK cells that have been modified by the hand of man to express part or all of a CD3 / TCR receptor complex. In specific embodiments, the NK cells are modified to include all components of the CD3 complex, including CD3^, CD3s, CD3y and CD36. In particular cases the full lengths of CD3^, CD3s, CD3y and CD36 are utilized, including their extracellular domain, transmembrane domain, and intracellular domain, however in alternative embodiments only part of one or more of CD3^, CD3s, CD3y and CD36 are utilized each of which that may or may not be combined with one or more intracellular signaling domains such as CD 16, NKG2D, DAP 10, DAP 12, CD28, 4 IBB, 2B4, CD27, 0X40, or any combination thereof.

[0179] In certain embodiments, an amino acid sequence (e.g., a polypeptide) may comprise an amino acid represented by a single letter “X” or a three letter code “Xaa”. In some embodiments, the amino acid represented by “X” or “Xaa” is any naturally occurring amino acid, such as but not limited to, Arginine (Arg, R), Histidine (His, H), Lysine (Lys, K), Aspartic Acid (Asp, D), Glutamic Acid (Glu, E), Serine (Ser, S), Threonine (Thr, T), Asparagine (Asn, N), Glutamine (Gin, Q), Glycine (Gly, G), Proline (Pro, P), Cysteine (Cys, C), Alanine (Ala, A), Valine (Vai, V), Isoleucine (He, I), Leucine (Leu, L), Methionine (Met, M), Phenylalanine (Phe, F), Tyrosine (Tyr, Y), or Tryptophan (Trp, W).

[0180] In certain embodiments, particular sequences for any of the CD3 receptor components are utilized, including wildtype or mutants of the components so long as the CD3 receptor having the mutant is able to allow signaling through the CD3 complex leading to activation and killing of targets. In some cases, CD3 / TCR complex associated polypeptides, polynucleotides encoding the same, and / or constructs comprising said polynucleotides are described in the Inventors international patent application publication W02023004425A2 (PCT / US2022 / 074062), published on January 26, 2023, which is incorporated herein by reference in its entirety for the purposes described herein.

[0181] In some cases, the following examples of sequences for CD3s, CD36, CD3y, and CD3^ and are utilized for modification of the NK cells.

[0182] CD3 Epsilon (CD3s, CD3e) (UniProtKB - P07766 (CD3E HUMAN))

[0183] Signal PeptideMQSGTHWRVLGLCLLSVGVW ( SEQ ID NO : 1 )

[0184] Extracellular Domain sp|P07766|23-126DGNEEMGGI TQTPYKVS I SGTTVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDHLSL KEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD ( SEQ ID NO : 2 )

[0185] Transmembrane Domain sp|P07766| 127-152VMSVAT IVIVDICI TGGLLLLVYYWS ( SEQ ID NO : 3 )

[0186] Intracellular Domain sp|P07766| 153-207KNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI ( SEQ ID NO : 4 )

[0187] An example of a Homo sapiens CD3e molecule (CD3E), mRNA is at NCBI Reference Sequence: GENBANK® Accession No. NM_000733.4 ATGCAGTCGGGCACTCACTGGAGAGTTCTGGGCCTCTGCCTCTTATCAGTTGGCGTTTGGGG GCAAGATGGTAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCTCTG GAAC GAG AG T AAT AT T GAG AT G C C C T GAG TATCCTGGATCT GAAAT AC T AT G G C AAC AC AAT GATAAAAACATAGGCGGTGATGAGGATGATAAAAACATAGGCAGTGATGAGGATCACCTGTC ACTGAAGGAATTTTCAGAATTGGAGCAAAGTGGTTATTATGTCTGCTACCCCAGAGGAAGCA AACCAGAAGATGCGAACTTTTATCTCTACCTGAGGGCAAGAGTGTGTGAGAACTGCATGGAG ATGGATGTGATGTCGGTGGCCACAATTGTCATAGTGGACATCTGCATCACTGGGGGCTTGCT GCTGCTGGTTTACTACTGGAGCAAGAATAGAAAGGCCAAGGCCAAGCCTGTGACACGAGGAG CGGGTGCTGGCGGCAGGCAAAGGGGACAAAACAAGGAGAGGCCACCACCTGTTCCCAACCCA GACTATGAGCCCATCCGGAAAGGCCAGCGGGACCTGTATTCTGGCCTGAATCAGAGACGCAT CTGA ( SEQ ID NO : 5 )

[0188] Examples of respective nucleic acid and amino acid CD3 epsilon sequences in their entirety are as follows (underlining refers to signal peptide sequence, which in some embodiments may be modified, omitted, and / or replaced with an alternative signal peptide): ATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGGG CCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGCG GCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAAC GACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGAG CCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGCA AGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGAG ATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGCT GCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGCG CCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCCC GACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAAT C ( SEQ ID NO : 6 )MQSGTHWRVLGLCLLSVGVWGQDGNE EMGG I T Q T P YKVS I S G T T V I L T C P Q Y P G S E I L WQHN DKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI ( SEQ ID NO : 7 )

[0189] CD3 Delta (CD38, CD3d) (UniProtKB - P04234 (CD3D HUMAN))

[0190] Signal PeptideMEHSTFLSGLVLATLLSQVS ( SEQ ID NO : 8 )

[0191] Extracellular Domain sp|P04234|22-105FKI PIEELEDRVFVNCNTS I TWVEGTVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKE STVQVHYRMCQSCVELDPATVA ( SEQ ID NO : 9 )

[0192] Transmembrane Domain sp|P04234| 106-126GI IVTDVIATLLLALGVFCFA ( SEQ ID NO : 10 )

[0193] Intracellular Domain sp|P04234|127-171GHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK ( SEQ ID NO : 11 )

[0194] Homo sapiens CD3d molecule, delta (CD3-TCR complex), mRNA (cDNA clone MGC:88324 IMAGE:30412345), complete cds GENBANK®: BC070321.1ATGGAACATAGCACGTTTCTCTCTGGCCTGGTACTGGCTACCCTTCTCTCGCAAGTGAGCCC C T T C AAGAT AC C T AT AGAG GAAC T T GAG GAG AGAG T G T T T G T GAAT T G C AAT AC C AG CAT C A CATGGGTAGAGGGAACGGTGGGAACACTGCTCTCAGACATTACAAGACTGGACCTGGGAAAA CGCATCCTGGACCCACGAGGAATATATAGGTGTAATGGGACAGATATATACAAGGACAAAGA ATCTACCGTGCAAGTTCATTATCGAATGTGCCAGAGCTGTGTGGAGCTGGATCCAGCCACCG TGGCTGGCATCATTGTCACTGATGTCATTGCCACTCTGCTCCTTGCTTTGGGAGTCTTCTGC TTTGCTGGACATGAGACTGGAAGGCTGTCTGGGGCTGCCGACACACAAGCTCTGTTGAGGAA TGACCAGGTCTATCAGCCCCTCCGAGATCGAGATGATGCTCAGTACAGCCACCTTGGAGGAA ACTGGGCTCGGAACAAGTGA ( SEQ ID NO : 12 )

[0195] Examples of respective nucleic acid and amino acid CD3 delta sequences in their entirety are as follows (underlining refers to signal peptide sequence, which in some embodiments may be modified, omitted, and / or replaced with an alternative signal peptide):ATGGAGCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCC CTTCAAGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCA CCTGGGTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAG AGAATCCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGA GAGCACCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCG TGGCCGGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGC TTCGCCGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAA CGACCAGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCA ACTGGGCCAGAAACAAG ( SEQ ID NO : 13 )MEHSTFLSGLVLATLLSQVSPFKI PIEELEDRVFVNCNTS I TWVEGTVGTLLSDI TRLDLGK RILDPRGI YRCNGTDI YKDKESTVQVHYRMCQSCVELDPATVAGI IVTDVIATLLLALGVFC FAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK ( SEQ ID NO : 14 )

[0196] CD3 Gamma (CD3y, CD3g) (T-cell surface glycoprotein CD3 gamma chain Gene CD3G P09693)Signal PeptideMEQGKGLAVL ILAI ILLQGTLA ( SEQ ID NO : 15 )

[0197] Extracellular Domain sp|P09693|23-l 16QS IKGNHLVKVYDYQEDGSVLLTCDAEAKNI TWFKDGKMIGFLTEDKKKWNLGSNAKDPRGM YQCKGSQNKSKPLQVYYRMCQNCIELNAAT I S ( SEQ ID NO : 1 6 )

[0198] Transmembrane Domain sp|P09693|l 17-137GFLFAE IVS I FVLAVGVYFIA ( SEQ ID NO : 17 )

[0199] Intracellular Domain sp|P096931138- 182GQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRN ( SEQ ID NO : 18 )

[0200] Homo sapiens CD3g molecule (CD3G), mRNA; NM_000073.3:81-629 Homo sapiens CD3g molecule (CD3G), mRNAATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTT GGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGG TAG T T C T GAG T T GT GAT GCAGAAGCCAAAAATAT CACAT GGT T TAAAGAT GGGAAGAT GAT C GGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGAGG GAT G TAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T GTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTC AGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTC GAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATC GAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATTGA ( SEQ ID NO : 19 )

[0201] Examples of respective nucleic acid and amino acid CD3 gamma sequences in their entirety are as follows (underlining refers to signal peptide sequence, which in some embodiments may be modified, omitted, and / or replaced with an alternative signal peptide): ATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTT GGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGG TAG T T C T GAC T T GT GAT GCAGAAGCCAAAAATAT CACAT GGT T TAAAGAT GGGAAGAT GAT C GGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGG GAT G TAT C AG T G T AAAG GAT C AC AGAAC AAG T C AAAAC C AC T C C AAG T G T AT TAG AGAAT G T GTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTC GAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATC GAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAAT ( SEQ ID NO : 20 )MEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMI GFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIV S I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRN ( SEQ ID NO : 21 )

[0202] CD3 Zeta (CD3 , CD3z)

[0203] Signal Peptide sp|P20963| SPMKWKALFTAAILQAQLPITEA ( SEQ ID NO : 22 )

[0204] Extracellular Domain sp|P20963122-30 ECDQSFGLLDPK ( SEQ ID NO : 23 )

[0205] Transmembrane Domain sp|P20963131-51 tmdLCYLLDGILFI YGVILTALEL ( SEQ ID NO : 24 )

[0206] Intracellular Domain sp|P20963|52-164 ICDRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNE LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO : 25 )

[0207] Examples of respective nucleic acid and amino acid CD3 zeta sequences in their entirety are as follows (underlining refers to signal peptide sequence, which in some embodiments may be modified, omitted, and / or replaced with an alternative signal peptide): ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCC GCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTA CGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGA AGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGT GAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT CAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGC ( SEQ ID NO : 26 )MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAP AYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYS EIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO : 27 )

[0208] Homo sapiens CD247 molecule (CD247; also referred to as CD3 Zeta), transcript variant 1, mRNANCBI Reference Sequence: NM_198053.3NM_198053.3:65-559 Homo sapiens CD247 molecule (CD247), transcript variant 1, mRNA ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCC GCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTA CGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGA AGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGT GAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT CAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCTAA ( SEQ ID NO : 28 )

[0209] In specific embodiments, the NK cells are modified to express one of more of the TCRa chain, the TCR0 chain, the TCRy chain, and the TCR5 chain, and any combination thereof may be utilized. In certain embodiments, the TCR may be an invariant Natural Killer cell TCR (iTCR). In a specific case, the NK cells are modified to express the T-cell receptor (TCR) 0.0 chains, iTCR a0 chains, or the TCR y5 chains. In certain cases, the NK cells are modified to express part or all of only the constant region of one of more of the TCRa chain, iTCRa chain, the TCR0 chain, iTCR0 chain, the TCRy chain, and the TCR5 chain. The NK cells may be modified to express part or all of only the constant region of the T-cell receptor (TCR) 0 chains, or the TCR y5 chains, or the iTCR 0 chains. In cases wherein part of the constant region is utilized, the part of the constant region may be at least 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, or 400 amino acids, including contiguous amino acids of any constant region. The part of the constant region may comprise at least 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the amino acids of a constant region, including contiguous amino acids of a constant region.

[0210] In specific cases, any sequences encompassed herein are utilized to modify the NK cells, although in other cases sequences that are related to these in identity are utilized. For example, related sequences that are at least 80, 85, 90, 95, 96, 97, 98, 99% identical to any sequence encompassed herein may be utilized in the disclosure.

[0211] Particular constructs for the expression of various TCR / CD3 proteins in the NK cells may be utilized, and in a variety of configurations. In specific cases, the NK cells may be transduced or transfected with one or more vectors to express any of the various proteins encompassed herein, including at least any one or more components of the TCR / CD3 complex. In specific cases, the one or more vectors themselves may or may not be multi ci str onic by beingable ultimately to produce more than one separate polypeptide. In cases wherein one or more multi ci str onic vectors are employed, they may utilize one or more internal ribosome entry sites (IRES) and / or one or more 2A self-cleaving peptide sites. In some embodiments, a 2A selfcleaving peptide site is encoded by a codon optimized polynucleotide. In cases wherein one or more 2A sequences are utilized, the following may be used, where GSG is an optional linker:

[0212] T2A ( GSG) EGRGSLLTCGDVEENPGP ( SEQ ID NO : 29 )

[0213] P2A ( GSG) ATNFSLLKQAGDVEENPGP ( SEQ ID NO : 30 )

[0214] E2A ( GSG) QCTNYALLKLAGDVESNPGP ( SEQ ID NO : 31 )

[0215] F2A ( GSG) VKQTLNFDLLKLAGDVESNPGP ( SEQ ID NO : 32 )

[0216] In situations wherein multiple protein components are expressed from a multi ci str onic vector, the order in a 5' to 3' direction on the polynucleotide vector may be of any order, although in alternative cases they are present on the vector in a particular order. A multi ci str onic vector may express multiple components of the TCR / CD3 receptor complex and no other heterologous protein, or the multi ci stronic vector may express multiple components of the TCR / CD3 receptor complex and one or more other heterologous proteins. In specific embodiments, two or more multicistronic vectors are provided, each encoding one or more components of the TCR / CD3 receptor complex and one or more other heterologous proteins, such as a cytokine and / or an engineered receptor (e.g., a receptor comprising an extracellular portion of an Fc binding protein).

[0217] There is an example in FIGs. 3A and 3C of a multicistronic vector in which full lengths of CD3s, CD36, CD3y, and CD3^ are present and separated by the same or different 2A self-cleaving peptide sites (“CD3 complex”). A multicistronic vector may include the signal peptide, extracellular domain, transmembrane domain, and intracellular domain of each of CD3s, CD35, CD3y, and CD3

[0218] FIG. 3C provides examples of various iTCR and / or CD 16 expression constructs for engineering of NK cells. In particular embodiments of the disclosure, TCR / CD3 receptor complex components are expressed from different vectors in the NK cells. In any case, the vector(s) may express a TCR directed against, or not directed against, a particular antigen of interest, such as a cancer antigen or a viral antigen. The TCR may or may not comprise at least part of CD3^, including the intracellular domain of CD3^, in addition to the NK cells also expressing CD3^ as a separate molecule from the TCR and as part of the CD3 receptor complex.

[0219] In specific embodiments, a TCR, such as an iTCR of the modified NK cells is utilized not necessarily as a therapeutic and / or targeting moiety aspect for the NK cells, but as a structural support or scaffold to facilitate function or enhanced function of the CD3 receptor complex. That is, the TCR may be any TCR and may not necessarily be utilized for its ability to target a particular antigen. In such cases, as non-limiting examples, a TCR that targets a viral antigen, a glycolipid, a bacterial antigen, etc. may be employed for NK cells that will be used for cancers that are not necessarily related to that particular virus, bacteria, and / or glycolipid. In other cases, the TCR is selected for the ability to target a particular antigen associated with a cancer of interest (e.g., a tumor associated antigen).

[0220] In certain embodiments, provided herein are CD3 constructs comprising a fusion with an intracellular co-stimulatory domain derived from CD 16, NKG2D, DAP 10, DAP 12, 2B4, 4- IBB, CD2, CD28, DNAM, or any combination thereof. In certain embodiments, an intracellular co-stimulatory domain is fused to CD36, CD3s, CD3y, and / or CD3<^. In certain embodiments, such a CD3 fusion construct comprises a CD3<^ fused to a DAP 10 intracellular co-stimulatory domain. In certain embodiments, such a CD3 fusion construct comprises a CD3<^ fused to a CD28 intracellular co-stimulatory domain. In certain embodiments, such a CD3 fusion construct comprises a CD3<^ fused to a DAP 10 intracellular co-stimulatory domain and a CD28 intracellular co-stimulatory domain. In certain embodiments, a CD3<^ fused to a DAP 10 intracellular co-stimulatory domain is represented by a 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 SEQ ID NO: 33. In certain embodiments, a CD3<^ fused to a CD28 intracellular co-stimulatory domain is represented by 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 SEQ ID NO: 34. In certain embodiments, a CD3<^ fused to a DAP10 intracellular co- stimulatory domain and a CD28 intracellular co-stimulatory domain is represented by 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 SEQ ID NO: 35. In certain embodiments, a CD3<^ fused to a DAP10 intracellular co- stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 36. In certain embodiments, a CD3<^ fused to a CD28 intracellular co-stimulatory domain is represented by an amino acid sequence thatis 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: 37. In certain embodiments, a CD3^ fused to a DAP10 intracellular co-stimulatory domain and a CD28 intracellular co-stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38. In certain embodiments, a CD3^ fused to an intracellular domain may not comprise a C terminal 2A domain. In certain embodiments, a CD3^ fused to an intracellular domain may not comprise an N terminal signal peptide domain.ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGCTTTGCGCACGCCCACGCCGCAGCCCCGCCCAA GAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGC AGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAA GAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCG CAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGA GGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTT ACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCC CCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGG CCCC ( SEQ ID NO : 33 )ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGAC TACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCAAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCG CGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTAC GATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAA GAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTG AGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTC AGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTG CACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC ( SEQ ID NO : 34 )ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGAC TACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCACTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAG AAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGCA GACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAG AGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGC AGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAG GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTA CCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCC ( SEQ ID NO : 35 )MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLLCARPRRSPAQ EDGKVYINMPGRGRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKP QRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALP PRQCTNYALLKLAGDVESNPGP ( SEQ ID NO : 36 )MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSD YMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGL STATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGP ( SEQ ID NO : 37 )MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSD YMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMPGRGRVKFSRSA DAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGP ( SEQ ID NO : 38 )

[0221] In certain embodiments, a DAP 10 intracellular co-stimulatory domain is represented by 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 SEQ ID NO: 39. In certain embodiments, a CD28 intracellular costimulatory domain is represented by 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 SEQ ID NO: 40. In certain embodiments, a DAP10 intracellular co-stimulatory domain and CD28 intracellular co-stimulatory domain is represented by 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 SEQ ID NO: 41. In certain embodiments, a DAP 10 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: 42. In certain embodiments, a CD28 intracellular co-stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 43. In certain embodiments, a DAP 10 intracellular co-stimulatory domain and CD28 intracellular co- stimulatory domain is represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 44.CTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGC ( SEQ ID NO : 39 )AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGG GCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCA ( SEQ ID NO : 40 )AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGG GCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCAC TTTGCGCACGCCCACGCCGCAGCCCCGCCCAAGAAGATGGCAAAGTCTACATCAACATGCCA GGCAGGGGC ( SEQ ID NO : 41 )LCARPRRSPAQEDGKVYINMPGRG ( SEQ ID NO : 42 )RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS ( SEQ ID NO : 43 )RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMP GRG ( SEQ ID NO : 44 )

[0222] UTNK15-D AP 10 : refers to full length CD3zeta comprising a fusion with an intracellular co-stimulatory domain derived from DAP10, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to IL 15, it may be represented by a 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 SEQ ID NO: 45. In certain embodiments, a UTNK15-DAP10 amino acid sequence may be represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46.ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGCTTTGCGCACGCCCACGCCGCAGCCCCGCCCAA GAAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGC AGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAA GAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCG CAGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGA GGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTT ACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCC CCTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGG CCCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTA CTTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGT TCGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGAT GATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTC GTGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT TAG AGA ATGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAAT CGTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAG GATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCA GACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGG AGCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTC AAGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTG GGTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAA TCCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGC ACCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGC CGGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCG CCGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGAC CAGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTG GGCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCG GCCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTG TGGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCAT CAGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGC ACAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCAC CTGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGG CAGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCA TGGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGC CTGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAG AGGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCA ACCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGA AGAATCGGACCGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAA TCCCGGGCCCATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGT GCCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGC TTCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAA GATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGC ACCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTG GAAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAG CCTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGA AGAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 45 )MKWKALFTAAILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLLCARPRRSPAQ EDGKVYINMPGRGRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKP QRRKNPQEGLYNELQKDKMAEAYSE IGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALP PRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDG SVLLTCDAEAKNI TWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYR MCQNCIELNAAT I SGFLFAE IVS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLK DREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPF KI PIEELEDRVFVNCNTS I TWVEGTVGTLLSDI TRLDLGKRILDPRGIYRCNGTDIYKDKES T VQVH YRMC Q S C VE L D PAT VAG I IVTDVIATLL LAL GVFC FAGHE T GRL S GAAD T QAL LRND QVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGV WGQDGNEEMGGI TQTPYKVS I SGTTVILTCPQYPGSE ILWQHNDKNIGGDEDDKNIGSDEDH LSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVAT IVIVDICI TGG LLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQR RIGPQCTNYALLKLAGDVESNPGPMRI SKPHLRS I S IQCYLCLLLNSHFLTEAGIHVFILGC FSAGLPKTEANWVNVI SDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVI SL ESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIKEFLQS FVHIVQMFINTS ( SEQ ID NO : 4 6 )

[0223] UTNK15-28: refers to full length CD3zeta comprising a fusion with an intracellular co-stimulatory domain derived from CD28, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to IL15, it may be represented by a 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 SEQ ID NO: 47. In certain embodiments, a UTNK 15-28 amino acid sequence may be represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 48.ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGC ACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCT ATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGAC TACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCAAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCG CGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTAC GATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGCAGAGAAGGAA GAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTG AGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTC AGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCCAGTG CACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAAC AGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTACTTTGGCCCAG TCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTTCGGTACTTCT GACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATGATCGGCTTCC TAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGTGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAAT G T G T C AGAA CTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTCAGCATTT TCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCAGTCGAGAGCT TCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGGATCGAGAAGA TGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAGACCCTGAACT TCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGAGCACAGCACC TTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCAAGATCCCCAT CGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGGGTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAATCCTGGACCCC AGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCACCGTGCAGGT GCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCCGGCATCATCG TGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGCCGGCCACGAG ACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACCAGGTGTACCA GCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGGGCCAGAAACA AGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGGCCCCATGCAG AGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGTGGGGCCAGGA CGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATCAGCGGCACCA CCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCACAACGACAAG AACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACCTGAGCCTGAA GGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGCAGCAAGCCCG AGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCATGGAGATGGAC GTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCCTGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGAGGCGCCGGCG CCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAACCCCGACTAC GAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAAGAATCGGACC GCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCA TGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTGCCTGCTGCTG AACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCTTCAGCGCCGG ACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAGATCGAGGACC TGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCACCCCAGCTGC AAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGGAAAGCGGCGA CGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGCCTGAGCAGCA ACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCAAA GAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 47 )MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSD YMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGL STATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPGPMEQGKGLAVLILAI ILLQGTLAQ S IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMY QCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVS I FVLAVGVYFIAGQDGVRQSRA SDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQTLNFDLLKLAGDVESNPGPMEHST FLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITWVEGTVGTLLSDITRLDLGKRILDP RGI YRCNGTDI YKDKESTVQVHYRMCQSCVELDPATVAGI IVTDVIATLLLALGVFCFAGHE TGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNKEGRGSLLTCGDVEENPGPMQ SGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS ISGTTVILTCPQYPGSEILWQHNDK NIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD VMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDY EPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESNPGPMRISKPHLRS IS IQCYLCLLL NSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSC KVTAMKCFLLELQVISLESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIK EFLQSFVHIVQMFINTS ( SEQ ID NO : 48 )

[0224] UTNK15-28-DAP10: refers to full length CD3zeta comprising a fusion with an intracellular co- stimulatory domain derived from DAP 10 and an intracellular co-stimulatory domain derived from CD28, full length CD3 gamma, full length CD3 delta, and full length CD3 epsilon linked to IL15, it may be represented by a 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 SEQ ID NO: 49. In certain embodiments, a UTNK15-28-DAP10 amino acid sequence may be represented by an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ IDNO: 50.ATGAAGTGGAAGGCGCTTTTCACCGCGGCCATCCTGCAGGCACAGTTGCCGATTACAGAGGCACAGAGCTTTGGCCTGCTGGATCCCAAACTCTGCTACCTGCTGGATGGAATCCTCTTCATCTATGGTGTCATTCTCACTGCCTTGTTCCTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCC ACCACGCGACTTCGCAGCCTATCGCTCACTTTGCGCACGCCCACGCCGCAGCCCCGCCCAAG AAGATGGCAAAGTCTACATCAACATGCCAGGCAGGGGCAGAGTGAAGTTCAGCAGGAGCGCA GACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAG AGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGC AGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAG GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTA CCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCC CTCGCCAGTGCACCAACTACGCCCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGC CCCATGGAACAGGGGAAGGGCCTGGCTGTCCTCATCCTGGCTATCATTCTTCTTCAAGGTAC TTTGGCCCAGTCAATCAAAGGAAACCACTTGGTTAAGGTGTATGACTATCAAGAAGATGGTT CGGTACTTCTGACTTGTGATGCAGAAGCCAAAAATATCACATGGTTTAAAGATGGGAAGATG ATCGGCTTCCTAACTGAAGATAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCG TGGGATGTAT GAG T G T AAAG GAT C AC AGAAC AAG T C AAAAC GAG T C C AAG T G T AT T AC AGAA TGTGTCAGAACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATC GTCAGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCCA GTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCCTCAAGG ATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGGAATGTGAAGCAG ACCCTGAACTTCGACCTGCTGAAGCTGGCCGGCGACGTGGAGAGCAACCCCGGCCCCATGGA GCACAGCACCTTCCTGAGCGGCCTGGTGCTGGCCACCCTGCTGAGCCAGGTGAGCCCCTTCA AGATCCCCATCGAGGAGCTGGAGGACAGAGTGTTCGTGAACTGCAACACCAGCATCACCTGG GTGGAGGGCACCGTGGGCACCCTGCTGAGCGACATCACCAGACTGGACCTGGGCAAGAGAAT CCTGGACCCCAGAGGCATCTACAGATGCAACGGCACCGACATCTACAAGGACAAGGAGAGCA CCGTGCAGGTGCACTACAGAATGTGCCAGAGCTGCGTGGAGCTGGACCCCGCCACCGTGGCC GGCATCATCGTGACCGACGTGATCGCCACCCTGCTGCTGGCCCTGGGCGTGTTCTGCTTCGC CGGCCACGAGACCGGCAGACTGAGCGGCGCCGCCGACACCCAGGCCCTGCTGAGAAACGACC AGGTGTACCAGCCCCTGAGAGACAGAGACGACGCCCAGTACAGCCACCTGGGCGGCAACTGG GCCAGAAACAAGGAGGGCAGAGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAGAACCCCGG CCCCATGCAGAGCGGCACCCACTGGAGAGTGCTGGGCCTGTGCCTGCTGAGCGTGGGCGTGT GGGGCCAGGACGGCAACGAGGAGATGGGCGGCATCACCCAGACCCCCTACAAGGTGAGCATC AGCGGCACCACCGTGATCCTGACCTGCCCCCAGTACCCCGGCAGCGAGATCCTGTGGCAGCA CAACGACAAGAACATCGGCGGCGACGAGGACGACAAGAACATCGGCAGCGACGAGGACCACC TGAGCCTGAAGGAGTTCAGCGAGCTGGAGCAGAGCGGCTACTACGTGTGCTACCCCAGAGGC AGCAAGCCCGAGGACGCCAACTTCTACCTGTACCTGAGAGCCAGAGTGTGCGAGAACTGCAT GGAGATGGACGTGATGAGCGTGGCCACCATCGTGATCGTGGACATCTGCATCACCGGCGGCC TGCTGCTGCTGGTGTACTACTGGAGCAAGAACAGAAAGGCCAAGGCCAAGCCCGTGACCAGA GGCGCCGGCGCCGGCGGCAGACAGAGAGGCCAGAACAAGGAGAGACCCCCCCCCGTGCCCAA CCCCGACTACGAGCCCATCAGAAAGGGCCAGAGAGACCTGTACAGCGGCCTGAACCAGAGAA GAATCGGACCGCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAAT CCCGGGCCCATGCGCATTAGCAAGCCCCACCTGCGGAGCATCAGCATCCAGTGCTACCTGTG CCTGCTGCTGAACAGCCACTTCCTGACCGAGGCCGGCATCCACGTGTTCATCCTGGGCTGCT TCAGCGCCGGACTGCCCAAGACCGAGGCCAACTGGGTGAACGTGATCAGCGACCTGAAGAAG ATCGAGGACCTGATCCAGAGCATGCACATCGACGCCACCCTGTACACCGAGAGCGACGTGCA CCCCAGCTGCAAGGTGACCGCCATGAAGTGCTTTCTGCTGGAACTGCAGGTGATCAGCCTGG AAAGCGGCGACGCCAGCATCCACGACACCGTGGAGAACCTGATCATCCTGGCCAACAACAGC CTGAGCAGCAACGGCAACGTGACCGAGAGCGGCTGCAAAGAGTGCGAGGAACTGGAAGAGAA GAACATCAAAGAGTTTCTGCAGAGCTTCGTGCACATCGTGCAGATGTTCATCAACACCAGC ( SEQ ID NO : 49 )MKWKALFTAAILQAQLPI TEAQS FGLLDPKLCYLLDGILFIYGVILTALFLRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSLCARPRRSPAQEDGKVYINMPGRGRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPRQCTNYALLKLAGDVESNPG PMEQGKGLAVLILAI ILLQGTLAQS IKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKM IGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEI VS I FVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRNVKQ TLNFDLLKLAGDVESNPGPMEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTS ITW VEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVA GI IVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNW ARNKEGRGSLLTCGDVEENPGPMQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVS I SGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRG SKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTR GAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRIGPQCTNYALLKLAGDVESN PGPMRISKPHLRS IS IQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKK IEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDAS IHDTVENLI ILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS ( SEQ ID NO : 50 )

[0225] As depicted in FIG. 3C and described above, the term “linked” refers to being present on the same polynucleotide vector and does not necessarily mean that the two polypeptides are expressed as one polypeptide. For example, a cytokine produced from a vector of the disclosure may ultimately be produced as a separate molecule from any one or more TCR / CD3 receptor complex components. Whereas, the term “fused” or “fusion” refers to two polypeptides that comprise a peptide bond conjoining the two molecules, i.e. that the two polypeptides are covalently bound by an amide bond and are not separated by a splitting element, such as a 2A element.

[0226] In some embodiments, a TCR construct comprises Human papilloma virus (HPV)- specific TCR chains. In some embodiments, a TCR construct comprising an HPV-specific TCR chains comprises TCR alpha and TCR beta chains that target the HPV 18 E6 protein, and / or HPV 18 E7 protein. In some embodiments, an HPV 18 E6 epitope is amino acids 121-135 and / or amino acids 77-91 of the HPV 18 E6 protein. In some embodiments, a TCR construct comprising an HPV-specific TCR chains comprises TCR alpha and TCR beta chains that target the HPV 18 E7 protein. In some embodiments, an HPV 18 E7 epitope is amino acids 11-19. In some embodiments, HPV-specific TCR sequences, TCR variable domain sequences, CDR sequences, and / or TCR constant domain sequences, are described in international patent application publications WO 2015 / 009604 Al, which is incorporated herein by reference for the purpose described herein.

[0227] In some embodiments, a TCR is an invariant natural killer T cell TCR (iTCR). In some embodiments, an iTCR may bind to CD Id expressing cells, such as certain cancer cells, monocytes, and / or macrophages that promote tumor growth. While not being limited by theory, CD 1 d molecules are not polymorphic, and thus in certain embodiments, recognition of antigenspresented by CD Id by transgenic iTCR bearing NK cells does not result in a host-graft mismatch. In certain embodiments, utilization of iTCRs in place of standard T cell derived TCRs can reduce comorbidities associated with MHC-1 / 2 mismatch.

[0228] In some embodiments, an iTCR complex comprises an alpha (a) chain iTCR (iTCRa). In some embodiments, a construct encoding an iTCRa comprises a polynucleotide 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: 51 (e.g., iTCRa associated with Va24-Jal8 in humans). In some embodiments, an iTCRa comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 52 (e.g., iTCRa associated with Va24-Jal8 in humans).

[0229] iTCRaATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGC ( SEQ ID NO : 51 )MKKHLTTFLVILWLYFYRGNGKNQVEQS PQSL I I LEGKNCTLQCNYTVS PFSNLRWYKQDTG RGPVSLTIMTFSENTKSNGRYTATLDADTKQSSLHITASQLSDSASYICWSDRGSTLGRLY FGRGTQLTVWPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLD MRSMDFKSNSAVAWSNKSDFACANAFNNS I IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQ NLSVIGFRILLLKVAGFNLLMTLRLWSS ( SEQ ID NO : 52 )

[0230] iTCRp

[0231] In some embodiments, an iTCR complex comprises a beta (P) chain iTCR (iTCRP). In some embodiments, a construct encoding an iTCRp comprises a polynucleotide 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: 53, 55, or 57. In some embodiments, an iTCRp comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 54, 56, 58.ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGGCAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGCAGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTACTTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTACCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCCAGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGC ( SEQ ID NO : 53 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMELHLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSELRALGPSSYNSPLHFGNGTRLTVTDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVK RKDF ( SEQ ID NO : 54 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCTCTAGTGAAGGCGGAGGACTCAAGTTGGCGAAAAACATCCAATACTTTGGTGCTGGCACCCGGCTTTCTGTCCTGGACCTGAACAAGGTGTTCCCTCCAGAGGTGGCCGTGTTCGAGCCTTCTGAGGCCGAGATCAGCCACACACAGAAAGCCACACTCGTGTGTCTGGCCACCGGCTTTTTCCCCGATCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTGCACAGCGGCGTCtGCACAGATCCCCAGCCTCTGAAAGAACAGCCCGCTCTGAACGACAGCCGGTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACCACTTCAGATGCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGATAGAGCCAAGCCTGTGACACAGATCGTGTCTGCCGAAGCCTGGGGCAGAGCCGATTGTGGCTTTACCAGCGTGTCATACCAGCAGGGCGTGCTGTCTGCCACCATCCTGTATGAGATTCTGCTGGGCAAAGCCACTCTGTACGCCGTGCTGGTGTCTGCCCTTGTGCTGATGGCCATGGTCAAGAGAAAG GACTTC ( SEQ ID NO : 55 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMELHLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSEGGGLKLAKNIQYFGAGTRLSVLDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRK DE ( SEQ ID NO : 56 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCTCTAGTGAATTCGCCAGCTCCGTGCGAGGGAATACTATC TACTTTGGTGAGGGATCTTGGCTTACGGTAGTAGACCTGAACAAGGTGTTCCCTCCAGAGGT GGCCGTGTTCGAGCCTTCTGAGGCCGAGATCAGCCACACACAGAAAGCCACACTCGTGTGTC TGGCCACCGGCTTTTTCCCCGATCACGTGGAACTGTCTTGGTGGGTCAACGGCAAAGAGGTG CACAGCGGCGTCtGCACAGATCCCCAGCCTCTGAAAGAACAGCCCGCTCTGAACGACAGCCG GTACTGTCTGAGCAGCAGACTGAGAGTGTCCGCCACCTTCTGGCAGAACCCCAGAAACCACT TCAGATGCCAGGTGCAGTTTTACGGCCTGAGCGAGAACGACGAGTGGACCCAGGATAGAGCC AAGCCTGTGACACAGATCGTGTCTGCCGAAGCCTGGGGCAGAGCCGATTGTGGCTTTACCAG CGTGTCATACCAGCAGGGCGTGCTGTCTGCCACCATCCTGTATGAGATTCTGCTGGGCAAAG CCACTCTGTACGCCGTGCTGGTGTCTGCCCTTGTGCTGATGGCCATGGTCAAGAGAAAGGAC TTC ( SEQ ID NO : 57 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSEFASSVRGNTI YFGEGSWLTWDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEV HSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRA KPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F ( SEQ ID NO : 58 )

[0232] In some embodiments, an NK cell comprises an iTCR complex beta (P) chain iTCR (iTCRP). In some embodiments, a construct encoding an iTCRp comprises a polynucleotide 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: 59, 61, 63, 65, 67, 69, 71, or 73. In some embodiments, an iTCRp comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 60, 62, 64, 66, 68, 70, 72, or 74.

[0233] In some embodiments, a construct encoding an iTCRp comprises a polynucleotide 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: 59, 65, 69, or 71. In some embodiments, an iTCRp comprises an amino acid sequence that is at least, or exactly, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical to SEQ ID NO: 60, 66, 70, or 72.ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCA GGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTT TGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGG GATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCT TTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTC AAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTG ACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATA CCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGT ACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC ( SEQ ID NO : 59 ; iTCRp clone 3 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCATGQGAQDTQYFGP GTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSG VCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 60 ; iTCR[3 clone 3 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGATGGGGTGGGGAGCAATCAGCCCCAGCATTTT GGTGATGGGACTCGACTCTCCATCCTAGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGC GGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcG CGACGGGATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCAT TCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTA TTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCA GATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAA CCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgt gTCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGA CTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 61 ; iTCR[3 clone 18 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSDGVGSNQPQHF GDGTRLS ILEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVH SGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAK PVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 62 ; iTCR[3 clone 18 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGAGGGGGCTGGAAACACCATATATTTTGGAGAG GGAAGTTGGCTCACTGTTGTAGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGCGGTTTT TGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGG GATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGA GTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCT TTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTC AAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgtgTCATA CCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGT ACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 63 ; iTCRp clone 24 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSEGAGNTIYFGE GSWLTWEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSG VCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF ( SEQ ID NO : 64 ; 1TCRP clone 24 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGACAGGGATAGCAATCAGCCCCAGCATTTTGGT GATGGGACTCGACTCTCCATCCTAGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGCGGT TTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGA CGGGATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCC GGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTG CCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGAT GTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCC GTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgtgTC ATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTC TGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 65 ; 1TCRP clone 51 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSDRDSNQPQHFG DGTRLS ILEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHS GVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF( SEQ ID NO : 66 ; 1TCRP clone 51 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCGTGGGTCCGGTACCCTCCTACAATGAGCAGTTCTTC GGGCCAGGGACACGGCTCACCGTGCTAGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGC GGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcG CGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCAT TCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTA TTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCA GATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAA CCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGA ATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC ( SEQ ID NO : 67 ; 1TCRP clone 56 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASVGPVPSYNEQFF GPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVH SGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAK PVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDS RG ( SEQ ID NO : 68 ; iTCRp clone 56 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGGGGTGACTAGCGCCTCCTACAATGAGCAGTTC TTCGGGCCAGGGACACGGCTCACCGTGCTAGAGGACCTCAAGAATGTGTTTCCGCCCGAAGT CGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCC TcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTG CATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAG GTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACT TCAGATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCT AAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATC AGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAG CGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGAT AGTAGGGGC ( SEQ ID NO : 69 ; 1TCRP clone 76 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSGVTSASYNEQF FGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEV HSGVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRA KPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG ( SEQ ID NO : 70 ; 1TCRP clone 76 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGGAGGAGGGGAGGAGACCCAGTACTTCGGGCCA GGCACGCGGCTCCTGGTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTT TGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGG GATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGA GTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCT TTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTC AAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTG ACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATA CCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGT ACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGC( SEQ ID NO : 71 ; 1TCRP clone 93 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSGGGEETQYFGP GTRLLVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSG VCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPV TQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG ( SEQ ID NO : 72 ; iTCRp clone 93 )ATGACCATCCGGCTGCTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGC CGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAAT GCTCCCAGACCATGGGCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTG CATCTGATCCACTACAGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAG CACCGTGTCCAGAATCCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCC ACACCAGCCAGTACCTGTGTGCCAGCAGTGCGCAGGGGGTCAGCGAAAAACTGTTTTTTGGC AGTGGAACCCAGCTCTCTGTCTTGGAGGACCTCAATAAGGTGTTTCCGCCCGAAGTCGCGGT TTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGA CGGGATTTttcCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCC GGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTG CCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGAT GTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCC GTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAgtgTC ATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTC TGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGACTTC ( SEQ ID NO : 73 ; iTCRp clone 96 )MTIRLLCYMGFYFLGAGLMEADIYQTPRYLVIGTGKKITLECSQTMGHDKMYWYQQDPGMEL HLIHYSYGVNSTEKGDLSSESTVSRIRTEHFPLTLESARPSHTSQYLCASSAQGVSEKLFFG SGTQLSVLEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHS GVCTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF( SEQ ID NO : 74 ; iTCRp clone 96 )

[0234] In some embodiments, a construct encoding an iTCRp chain comprises an iTCRp V -DJ region comprising 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 SEQ ID NOs: 75-149.ACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGA G ( SEQ ID NO : 75 ; iTCRp clone 3 )AGCAGTGCCCCTGGAGGGTCTGAAGCTTTCTTTGGACAAGGCACCAGACTCACAGTTGTAGA G ( SEQ ID NO : 76 ; iTCR clone 4 )AGCAGTGAACTCGACAGGGAAGGAAACACCATATATTTTGGAGAGGGAAGTTGGCTCACTGT TGTAGAG ( SEQ ID NO : 77 ; iTCRp clone 7 )AGCAGTGATATGGGACCCGTCTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG ( SEQ ID NO : 78 ; iTCRp clone 9 )AGCAGTGAGGAAGAGCGGGGCCCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 79; iTCRp clone 13)AGCAGTGATGGGGTGGGGAGCAATCAGCCCCAGCATTTTGGTGATGGGACTCGACTCTCCAT CCTAGAG (SEQ ID NO: 80; 1TCRP clone 18)AGCAGTGACCCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG ( SEQ ID NO: 81; iTCR[3 clone 20)AGCAGTGAGGCCCCAACAGGAACCGGGGCCAACGTCCTGACTTTCGGGGCCGGCAGCAGGCT GACCGTGCTGGAG (SEQ ID NO: 82; iTCR[3 clone 21)AGCAGTGATCCTCGACTAGCGGGGGGGGTCGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG (SEQ ID NO: 83; iTCR[3 clone 23)AGCAGTGAGGGGGCTGGAAACACCATATATTTTGGAGAGGGAAGTTGGCTCACTGTTGTAGA G (SEQ ID NO: 84; iTCR[3 clone 24)AGCAGTTATGCTACAGGCTACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 85; iTCR[3 clone 26)AGCAGTGAACGGCAGGGCTCCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 86; iTCR[3 clone 28)AGCAGTGCTCCGACTAGCGGGAGGGACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCAC CGTGCTAGAG (SEQ ID NO: 87; iTCR[3 clone 29)AGCAGTGAATGGACTAGCGGGGGGCCCAACACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAG GCTGACCGTACTGGAG (SEQ ID NO: 88; iTCR[3 clone 30)AGCAGTGAACGGGGGGCTAGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 89; iTCRp clone 32)AGCAGTGAAGGGCTAGCGGGAGAACCTCTCTTAGGCAATGAGCAGTTCTTCGGGCCAGGGAC ACGGCTCACCGTGCTAGAG (SEQ ID NO: 90; iTCRp clone 33)AGCAGTGAGGCAGGCGGCCACACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 91; iTCRp clone 34)AGCAGTGAATACCAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGAG (SEQ ID NO: 92; iTCRp clone 36)AGCACCGACAGGGGATCTTTCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACT GGAG (SEQ ID NO: 93; iTCRp clone 37)GGAGGAGGGACATCTCAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGA G (SEQ ID NO: 94; iTCRp clone 38)AGCAGTCCGACTAGCGGGATGGGGGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGT GCTCGAG (SEQ ID NO: 95; iTCRp clone 39)AGCAGTGAGTTCGGGGCCAACGTCCTGACTTTCGGGGCCGGCAGCAGGCTGACCGTGCTGGA G (SEQ ID NO: 96; iTCRp clone 41)AGCAGTGTCCGTAGCGGGAGAGGGGACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCAC CGTGCTAGAG (SEQ ID NO: 97; 1TCRP clone 42)AGCAGTGTCCAGGAGGAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGA G (SEQ ID NO: 98; iTCR[3 clone 43)AGCAGTGATAGTAGCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG (SEQ ID NO: 99; iTCR[3 clone 44)AGCAGTGGTACTACGGGACAGGAATCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG (SEQ ID NO: 100; 1TCRP clone 46)AGCAGTGTAAGGGGGAACCACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 101; 1TCRP clone 47)AGCAGTGAACTTCAGCGGGAGGGTTCTCCAGATACGCAGTATTTTGGCCCAGGCACCCGGCT GACAGTGCTCGAG (SEQ ID NO: 102; 1TCRP clone 48)AGCAGTGTCCGGGACAGGGATGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTT GGAG (SEQ ID NO: 103; iTCR[3 clone 49)AGCAGTGAGGGTCAGGGAGGTTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCAC AGAG (SEQ ID NO: 104; iTCR[3 clone 50)AGCAGTGACAGGGATAGCAATCAGCCCCAGCATTTTGGTGATGGGACTCGACTCTCCATCCT AGAG (SEQ ID NO: 105; iTCR[3 clone 51)AGCAGTGATCGGTCTAGCGGAGCCAAAAACATTCAGTACTTCGGCGCCGGGACCCGGCTCTC AGTGCTGGAG (SEQ ID NO: 106; iTCR[3 clone 52)AGCAGTGCCACGACTAGCGGGAGGACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGAC AGTGCTGGAG (SEQ ID NO: 107; iTCRp clone 53)AGCAGTGAATTTCGGCAGCGGGAGTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG (SEQ ID NO: 108; iTCRp clone 54)AGCAGTGAAATAGCGGGAGTGGCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 109; iTCRp clone 55)AGCGTGGGTCCGGTACCCTCCTACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGT GCTAGAG (SEQ ID NO: 110; iTCRp clone 56)AGCAGTGAACGGCGCGGGAGACGGGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 111; iTCRp clone 57)AGCAGTGGGACAGGGTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG (SEQ ID NO: 112; iTCRp clone 58)AGCAGTGACCGTAGCGGGAGCTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 113; iTCR[3 clone 59)AGCAGTGACAGCACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACTGGAG (SEQ ID NO: 114; iTCRp clone 60)AGCAGTGCTAGCGGGAGCAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCTAGA G (SEQ ID NO: 115; iTCR[3 clone 61)AGCAGTGACGGGACTAGCGGCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCAC AGAG (SEQ ID NO: 116; 1TCRP clone 62)AGCAGTGAATATGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTTGGAG ( SEQ ID NO: 117; 1TCRP clone 63)AGCAGTGAGTCCGGCCCCCGCAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 118; 1TCRP clone 64)AGCAGTGGCCGACTAGCGGGAGAGGAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCT GGTGCTCGAG (SEQ ID NO: 119; 1TCRP clone 66)AGCAGTGAGGGTGGCAGGGTCGATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 120; 1TCRP clone 67)AGCAGTGAGGCTAACTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGTAGAG (SEQ ID NO: 121; iTCR[3 clone 68)AGCAGTCAGGACGGATTGGGATATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGT AGAG (SEQ ID NO: 122; iTCR[3 clone 69)AGCAGTGGGCGCCTCCACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG (SEQ ID NO: 123; iTCR[3 clone 70)AGCAGTGAATATAACAGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCT CGAG (SEQ ID NO: 124; iTCR[3 clone 71)AGCAGTGAACCCGGATTGGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACTGGA G (SEQ ID NO: 125; iTCR[3 clone 72)AGCATCCTGGGAGAGGGGCGGAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 126; iTCR[3 clone 73)AGCAGTGCCCCGGGACAGATCTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGT AGAG (SEQ ID NO: 127; iTCR[3 clone 74)AGCAGTGACAACCAAGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGAG (SEQ ID NO: 128; iTCR[3 clone 75)AGCAGTGGGGTGACTAGCGCCTCCTACAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCAC CGTGCTAGAG (SEQ ID NO: 129; iTCR[3 clone 76)AGCAGTCCTGAGCCCACCACCCTAGCGGGAGTCCACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGTCACAGAG (SEQ ID NO: 130; iTCRp clone 77)AGCAGTGGGACACAGAGGGCTGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTT GGAG (SEQ ID NO: 131; iTCR[3 clone 78)AGCAGTGGGACTAGCGGGAGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 132; 1TCRP clone 79)AGCAGTGAGGCGGGACAGGGTTCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 133; 1TCRP clone 80)AGCACCTCTAGCCGCACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACTGGA G (SEQ ID NO: 134; iTCR[3 clone 83)AGCAGTGAACCGGGGGAGCGGAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCT AGAG (SEQ ID NO: 135; iTCR[3 clone 84)AGCAGTGAAGGTCGGGTTAACTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGT AGAG (SEQ ID NO: 136; iTCR[3 clone 85)AGCAGTGAATCAGAAGGGGGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGT GCTCGAG (SEQ ID NO: 137; iTCRp clone 86)AGCAGTCCCGGGGGGACTAGCGGGAGGGCACGTCCCTACGAGCAGTACTTCGGGCCGGGCAC CAGGCTCACGGTCACAGAG (SEQ ID NO: 138; iTCRp clone 87)AGCAGTGGGAGGGAGGGGGACCCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 139; iTCRp clone 88)AGCAGTGGACTAGCGAACACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGACCGTACT GGAG (SEQ ID NO: 140; iTCRp clone 89)AGCAGTGGGACGACAGGGGATACACGCACAGATACGCAGTATTTTGGCCCAGGCACCCGGCT GACAGTGCTCGAG (SEQ ID NO: 141; iTCRp clone 90)AGCAGTGAAGACCGGGACAGGGGTCACGAGCAGTACTTCGGGCCGGGCACCAGGCTCACGGT CACAGAG (SEQ ID NO: 142; iTCRp clone 91)AGCAGTGAACTAGCGAATGAGCAGTTCTTCGGGCCAGGGACACGGCTCACCGTGCTAGAG (SEQ ID NO: 143; iTCRp clone 92)AGCAGTGGAGGAGGGGAGGAGACCCAGTACTTCGGGCCAGGCACGCGGCTCCTGGTGCTCGA G (SEQ ID NO: 144; iTCRp clone 93)AGCAGTGAATATGCAGGGTGGGGCGGCAATCAGCCCCAGCATTTTGGTGATGGGACTCGACT CTCCATCCTAGAG (SEQ ID NO: 145; iTCRp clone 94)AGCAGTGAATTGGACGGGACTAGCGCCTACGAGCAGTACTTCGGGCCGGGCACCAGGCTCAC GGTCACAGAG (SEQ ID NO: 146; iTCRp clone 95)AGCAGTGCGCAGGGGGTCAGCGAAAAACTGTTTTTTGGCAGTGGAACCCAGCTCTCTGTCTTGGAG ( SEQ ID NO : 147 ; iTCRp clone 96 )AGCAGTGAAGTGGCGGGAGCGGACACCGGGGAGCTGTTTTTTGGAGAAGGCTCTAGGCTGAC CGTACTGGAG ( SEQ ID NO : 148 ; iTCRp clone 97 )AGCAGCGGCAGGGGGCCAGGGGAAAGTGCAGATACGCAGTATTTTGGCCCAGGCACCCGGCT GACAGTGCTCGAG ( SEQ ID NO : 149 ; iTCRp clone 98 )B. NK Cell Fc Binding Modifications

[0235] As described herein, engineered NK cells can be modified to express a transgenic construct (heterologous construct) encoding a polypeptide comprising an Fc Receptor extracellular Fc binding domain (e.g., an Fc binding domain). The Fc binding domain can be tethered to a cell membrane, such as through a transmembrane domain (TMD). The Fc binding domain is comprised as part of, or obtained from, an Fc Receptor. An Fc receptor can be an Fey receptor, e g., FcyRI (CD64); FcyRIIA, IIB, and IIC (CD32); FcyRIIIA and / or IIIB (CD16). In certain embodiments, an Fey receptor is FcyRIII. In certain embodiments, an FcyRIII is FcyRIIIA, which can be a high affinity variant CD16 (CD16ha; e.g., F158V). In some embodiments, a cell expresses more than one transgenic Fc binding domain comprising polypeptides.

[0236] As described herein, a TCR / FcR construct comprises a coding sequence for an Fc Receptor extracellular binding domain. In exemplary embodiments described herein, a TCR / FcR construct can comprise chimeric polypeptides comprising an Fc binding domain derived from CD16 (e.g., FcyRIII). In some embodiments, an extracellular Fc binding domain derived from CD16 is a high affinity polymorph comprising an F158V substitution. In some embodiments, a CD16 derived Fc binding domain comprising polypeptide comprises substantially no intracellular signaling domain (ICD). In some embodiments, a CD 16 extracellular domain (e.g., Fc binding domain) comprising polypeptide comprises an optional heterologous hinge, a heterologous or CD 16 derived TMD, and / or a heterologous and / or CD 16 derived ICD. In certain embodiments, a CD16 derived Fc binding domain does not comprise mutations which render the binding domain uncleavable and / or refractory to cleavage.

[0237] In some embodiments, a polynucleotide encoding a transgenic Fc binding domain comprises a signal peptide. In some embodiments, a signal peptide is a signal peptide that can be found associated with the endogenous Fc Receptor that comprises the noted Fc binding domain. In some embodiments, a signal peptide is a heterologous signal peptide that is not naturally associated with an Fc binding protein.

[0238] In some embodiments, a TCR / FcR construct comprises an Fc Receptor extracellular binding domain polypeptide that is encoded by 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 SEQ ID NO: 150-154. In certain embodiments, a TCR / FcR construct comprises an Fc Receptor extracellular binding domain polypeptide 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: 155-159. In some embodiments, in sequences described herein, underlining refers to signal peptide sequence, which in some embodiments may be modified, omitted, and / or replaced with an alternative signal peptide.

[0239] As described herein, in some embodiments a TCR / FcR construct comprises a chimeric Fc Receptor extracellular binding domain polypeptide (e.g., that comprises one or more sequences derived from a non-Fc Receptor polypeptide and / or a different Fc Receptor polypeptide). In some embodiments, a TCR / FcR construct comprises a human CD32 derived hinge region linking an FC Receptor extracellular domain (ECD) sequence and a transmembrane domain (TMD) sequence. In some embodiments, a TCR / FcR construct comprises a CD3(^ derived TMD sequence and / or intracellular domain (ICD) sequence. In some embodiments, a human CD32 derived hinge region is encoded by 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 SEQ ID NO: 160. In certain embodiments, a human CD32 derived hinge region comprises a polypeptide 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: 161. In some embodiments, a CD3(^ derived TMD region is encoded by 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 SEQ ID NO: 162. In certain embodiments, a human CD3(^ derived TMD region comprises a polypeptide 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: 163. In some embodiments, a CD3(^ derived ICD region is encoded by 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 SEQ ID NO: 164. In certain embodiments, a human CD3(^ derived ICD region comprises a polypeptide 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: 165. In some embodiments, a TCR / FcR construct comprises a CD16 derived TMD sequence and / or ICD sequence. In some embodiments, a human CD 16 derived TMD region is encoded by 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 SEQ ID NO: 166. In certain embodiments, a human CD 16 derived TMD region comprises a polypeptide 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: 167. In some embodiments, a human CD16 derived ICD region is encoded by 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 SEQ ID NO: 168. In certain embodiments, a human CD16 derived ICD region comprises a polypeptide 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: 169.TCR / FcR #1 CD16 extracellular domain (Fc binding domain) comprising polypeptide: WT CD16, polynucleotide sequence (SEQ ID NO: 150)ATGTGGCAGCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GA AGATCTCCCAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACA GTGTGACTCTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCAC AATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGA CAGTGGAGAGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAG TCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATT CACCTGAGGTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGG CAAAGGCAGGAAGTAT T T T CAT CATAAT T C T GAG T T C TACAT T CCAAAAGCCACAC T CAAAG ACAGCGGCTCCTACTTCTGCAGGGGGCTTTTTGGGAGTAAAAATGTGTCTTCAGAGACTGTG AACATCACCATCACTCAAGGTTTGGCAGTGTCAACCATCTCATCATTCTTTCCACCTGGGTA CCAAgtctctttctgcttggtgatggtactcctttttgcagtggacacaggactatatttct ctgtgaagacaaacattcgaagctcaacaagagactggaaggaccataaatttaaatggaga aaggaccctcaagacaaaTAA ( SEQ ID NO : 150 )TCR / FcR #2 CD16 extracellular domain (Fc binding domain) comprising polypeptide: WT high affinity CD16, polynucleotide sequence (CD16ha, F158V) (SEQ ID NO: 151) ATGTGGCAGCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GA AGATCTCCCAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACA GTGTGACTCTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCAC AATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGA CAGTGGAGAGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAG TCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATT CACCTGAGGTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGG CAAAGGCAGGAAGTAT T T T CAT CATAAT T C T GAC T T C TACAT T CCAAAAGCCACAC T CAAAGACAGCGGCTCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTG AACATCACCATCACTCAAGGTTTGGCAGTGTCAACCATCTCATCATTCTTTCCACCTGGGTA CCAAgtctctttctgcttggtgatggtactcctttttgcagtggacacaggactatatttct ctgtgaagacaaacattcgaagctcaacaagagactggaaggaccataaatttaaatggaga aaggaccctcaagacaaaTAA ( SEQ ID NO : 151 )TCR / FcR #3 CD16 extracellular domain (Fc binding domain) comprising polypeptide: CD16ha ECD, CD32 hinge, CD16 TMD, and CD16 ICD, polynucleotide sequence (SEQID NO: 152)ATGTGGCAGCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GA AGATCTCCCAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACA GTGTGACTCTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCAC AATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGA CAGTGGAGAGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAG TCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATT CACCTGAGGTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGG CAAAGGCAGGAAGTAT T T T CAT CATAAT T C T GAG T T C TACAT T CCAAAAGCCACAC T CAAAG ACAGCGGCTCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTG AACATCACCGTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGGgtctctttctgctt ggtgatggtactcctttttgcagtggacacaggactatatttctctgtgaagacaaacattc gaagctcaacaagagactggaaggaccataaatttaaatggagaaaggaccctcaagacaaa TAA ( SEQ ID NO : 152 )TCR / FcR #4 CD16 extracellular domain (Fc binding domain) comprising polypeptide: CD16ha ECD, CD32 hinge, CD3 TMD, and CD3 ICD, polynucleotide sequence (SEQID NO: 153)ATGTGGCAGCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GA AGATCTCCCAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACA GTGTGACTCTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCAC AATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGA CAGTGGAGAGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAG TCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATT CACCTGAGGTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGG CAAAGGCAGGAAGTAT T T T CAT CATAAT T C T GAC T T C TACAT T CCAAAAGCCACAC T CAAAG ACAGCGGCTCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTG AACATCACCGTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGGCTCTGTTACCTTCT TGACGGTATTCTTTTTATTTACGGCGTCATCCTCACTGCCCTCTTTTTGAGGGTCAAGTTTT CAAGATCCGCCGACGCACCTGCTTACCAGCAAGGACAAAATCAACTTTATAACGAACTCAAC CTTGGTCGGCGGGAAGAATATGATGTACTCGACAAAAGAAGAGGCCGCGACCCGGAGATGGG TGGCAAACCACAGCGGAGGAAAAACCCCCAGGAAGGTCTCTATAACGAATTGCAGAAAGATA AAATGGCCGAGGCTTATTCAGAGATCGGCATGAAAGGAGAGCGACGACGAGGAAAGGGGCAT GATGGTCTGTATCAAGGGCTCAGCACGGCAACAAAGGACACATATGATGCCTTGCATATGCA GGCGCTTCCGCCGCGCTAA ( SEQ ID NO : 153 )TCR / FcR #5 CD16 extracellular domain (Fc binding domain) comprising polypeptide:CD16ha ECD, CD32 hinge, CD16 TMD, polynucleotide sequence (SEQ ID NO: 154)ATGTGGCAGCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GA AGATCTCCCAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACA GTGTGACTCTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCAC AATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGA CAGTGGAGAGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAG TCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATT CACCTGAGGTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGG CAAAGGCAGGAAGTAT T T T CAT CATAAT T C T GAG T T C TACAT T CCAAAAGCCACAC T CAAAG ACAGCGGCTCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTG AACATCACCGTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGGgtctctttctgctt ggtgatggtactcctttttgcagtggacacaggactatatttctctgtgaagacaTAA ( SEQ ID NO : 154 )TCR / FcR #1 CD16 extracellular domain (Fc binding domain) comprising polypeptide:WT CD16, amino acid sequence (SEQ ID NO: 155)MWQLLLPTALLLLVSAGMRTEDLPKAWFLEPQWYRVLEKDSVTLKCQGAYSPEDNSTQWFH NESLI SSQASSYFIDAATVDDSGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRWVFKEEDPI HLRCHSWKNTALHKVTYLQNGKGRKYFHHNSDFYI PKATLKDSGSYFCRGLFGSKNVSSETV NI T I TQGLAVS T I S S FFPPGYQVS FCLVMVLLFAVDTGLYFSVKTNIRS S TRDWKDHKFKWR KDPQDK ( SEQ ID NO : 155 )TCR / FcR #2 CD16 extracellular domain (Fc binding domain) comprising polypeptide:WT high affinity CD16, amino acid sequence (CD16ha, F158V) (SEQ ID NO: 156)MWQLLLPTALLLLVSAGMRTEDLPKAWFLEPQWYRVLEKDSVTLKCQGAYSPEDNSTQWFH NESLI SSQASSYFIDAATVDDSGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRWVFKEEDPI HLRCHSWKNTALHKVTYLQNGKGRKYFHHNSDFYI PKATLKDSGSYFCRGLVGSKNVSSETV NI T I TQGLAVS T I S S FFPPGYQVS FCLVMVLLFAVDTGLYFSVKTNIRS S TRDWKDHKFKWR KDPQDK ( SEQ ID NO : 156 )TCR / FcR #3 CD16 extracellular domain (Fc binding domain) comprising polypeptide: CD16ha ECD, CD32 hinge, CD16 TMD, and CD16 ICD, amino acid sequence (SEQ ID NO: 157)MWQLLLPTALLLLVSAGMRTEDLPKAWFLEPQWYRVLEKDSVTLKCQGAYSPEDNSTQWFH NESLI SSQASSYFIDAATVDDSGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRWVFKEEDPI HLRCHSWKNTALHKVTYLQNGKGRKYFHHNSDFYI PKATLKDSGSYFCRGLVGSKNVSSETV NI TVQVPSMGSSSPMGVS FCLVMVLLFAVDTGLYFSVKTNIRSSTRDWKDHKFKWRKDPQDK ( SEQ ID NO : 157 )TCR / FcR #4 CD16 extracellular domain (Fc binding domain) comprising polypeptide: CD16ha ECD, CD32 hinge, CD3 TMD, and CD3 ICD, amino acid sequence (SEQ ID NO: 158)MWQLLLPTALLLLVSAGMRTEDLPKAWFLEPQWYRVLEKDSVTLKCQGAYSPEDNSTQWFH NESLI SSQASSYFIDAATVDDSGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRWVFKEEDPIHLRCHSWKNTALHKVTYLQNGKGRKYFHHNSDFYIPKATLKDSGSYFCRGLVGSKNVSSETV NITVQVPSMGSSSPMGLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELN LGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO : 158 )TCR / FcR #5 CD16 extracellular domain (Fc binding domain) comprising polypeptide:CD16ha ECD, CD32 hinge, CD16 TMD, amino acid sequence (SEQ ID NO: 159)MWQLLLPTALLLLVSAGMRTEDLPKAWFLEPQWYRVLEKDSVTLKCQGAYSPEDNSTQWFH NESLISSQASSYFIDAATVDDSGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRWVFKEEDPIHLRCHSWKNTALHKVTYLQNGKGRKYFHHNSDFYIPKATLKDSGSYFCRGLVGSKNVSSETVNITVQVPSMGSSSPMGVSFCLVMVLLFAVDTGLYFSVKT ( SEQ ID NO : 159 )CD32 hinge polynucleotide (SEQ ID NO: 160)GTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGG ( SEQ ID NO : 160 )CD32 hinge polypeptide (SEQ ID NO: 161)VQVPSMGSSSPMG ( SEQ ID NO : 161 )CD3 transmembrane domain polynucleotide (SEQ ID NO: 162)CTCTGTTACCTTCTTGACGGTATTCTTTTTATTTACGGCGTCATCCTCACTGCCCTCTTTTT G ( SEQ ID NO : 162 )CD3 transmembrane domain polypeptide (SEQ ID NO: 163)LCYLLDGILFI YGVILTALEL ( SEQ ID NO : 163 )CD3 intracellular signaling domain polynucleotide (SEQ ID NO: 164)AGGGTCAAGTTTTCAAGATCCGCCGACGCACCTGCTTACCAGCAAGGACAAAATCAACTTTA TAACGAACTCAACCTTGGTCGGCGGGAAGAATATGATGTACTCGACAAAAGAAGAGGCCGCG ACCCGGAGATGGGTGGCAAACCACAGCGGAGGAAAAACCCCCAGGAAGGTCTCTATAACGAATTGCAGAAAGATAAAATGGCCGAGGCTTATTCAGAGATCGGCATGAAAGGAGAGCGACGACG AGGAAAGGGGCATGATGGTCTGTATCAAGGGCTCAGCACGGCAACAAAGGACACATATGATG CCTTGCATATGCAGGCGCTTCCGCCGCGCTAA ( SEQ ID NO : 164 )CD3 intracellular signaling domain polypeptide (SEQ ID NO: 165)RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNE LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO : 165 )CD16 transmembrane domain polynucleotide (SEQ ID NO: 166)GTCTCTTTCTGCTTGGTGATGGTACTCCTTTTTGCAGTGGACACAGGACTATATTTCTCTGT G ( SEQ ID NO : 166 )CD16 transmembrane domain polypeptide (SEQ ID NO: 167)VSFCLVMVLLFAVDTGLYFSV ( SEQ ID NO : 167 )CD16 intracellular signaling domain polynucleotide (SEQ ID NO: 168)AAGACAAACAT T CGAAGC T CAACAAGAGAC T GGAAGGACCATAAAT T TAAAT GGAGAAAGGA CCCTCAAGACAAATAA ( SEQ ID NO : 168 )CD16 intracellular signaling domain polypeptide (SEQ ID NO: 169)KTNIRSSTRDWKDHKFKWRKDPQDK ( SEQ ID NO : 169 )C. Exemplary Transgenic Open Reading Frames and Vectors

[0240] NK cells are modified to express a heterologous (i.e., transgenic) polynucleotide vector (i.e., construct). In some embodiments, a TCR / FcR construct comprises a multi ci stronic open reading frame 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 SEQ ID NO: 171-175. In certain embodiments, a TCR / FcR construct comprises a transgenic vector sequence book-ended by LTRs that is at least, or exactly, 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% identical to SEQ ID NO: 177-181. In some embodiments, in sequences described herein, underlining refers to signal peptide sequence, which in some embodiments may be modified, omitted, and / or replaced with an alternative signal peptide.

[0241] In some embodiments, provided herein are heterologous polynucleotide vectors comprising a pair of iTCR chains, while not comprising a CD16 extracellular domain (Fc binding domain) comprising polypeptide (e.g., an “iTCR” construct, e.g., construct iTCR3). In some embodiments, an iTCR construct comprises a multi ci stronic open reading frame 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 SEQ ID NO: 170. In certain embodiments, an iTCR construct comprises a transgenic vector sequence book-ended by LTRs that is at least, or exactly, 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% identical to SEQ ID NO: 176. iTCR3 open reading frame (SEQ ID NO: 170)ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATTTCAGCCTGCTGAAACAGGCTG GCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTGCTGTGCTATATGGGCTTCTAC TTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGG CACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGT ATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACC GAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACT GACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGG CGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAAT GTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAA GGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGG TTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCC GCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCA AAAC C C AAGAAAT GAG T T C AGAT G T C AAG T T GAG TTCTACGGTCT GAG C GAGAAT GAT GAG T GGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCG GATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGA AATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGCTAA ( SEQ ID NO : 170 )TCR / FcR #1 open reading frame (ORF) (SEQ ID NO: 171)ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATTTCAGCCTGCTGAAACAGGCTG GCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTGCTGTGCTATATGGGCTTCTAC TTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGG CACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGT ATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACC GAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACT GACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGG CGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAAT GTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAA GGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCC GCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCA AAAC C C AAGAAAT GAG T T C AGAT G T C AAG T T GAG TTCTACGGTCT GAG C GAGAAT GAT GAG T GGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCG GATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGA AATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAA TGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGTACTAATTATGCTCTCTTGAAA TTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCAGCTGCTCCTCCCAACTGCTCT GCTACTTCTAGTTTCAGCTGGCATGCGGACTGAAGATCTCCCAAAGGCTGTGGTGTTCCTGG AGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACTCTGAAGTGCCAGGGAGCCTAC TCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAGCCTCATCTCAAGCCAGGCCTC GAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAGAGTACAGGTGCCAGACAAACC TCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATCGGCTGGCTGTTGCTCCAGGCC CCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAGGTGTCACAGCTGGAAGAACAC TGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCAGGAAGTATTTTCATCATAATT CTGACTTCTACATTCCAAAAGCCACACTCAAAGACAGCGGCTCCTACTTCTGCAGGGGGCTT TTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCACCATCACTCAAGGTTTGGCAGT GTCAACCATCTCATCATTCTTTCCACCTGGGTACCAAgtctctttctgcttggtgatggtac tcctttttgcagtggacacaggactatatttctctgtgaagacaaacattcgaagctcaaca agagactggaaggaccataaatttaaatggagaaaggaccctcaagacaaaTAA ( SEQ ID NO : 171 )TCR / FcR #2 open reading frame (SEQ ID NO: 172)ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATTTCAGCCTGCTGAAACAGGCTG GCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTGCTGTGCTATATGGGCTTCTAC TTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGG CACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGT ATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACC GAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACT GACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGG CGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAAT GTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAA GGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGG TTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCC GCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCA AAAC C C AAGAAAT GAG T T C AGAT G T C AAG T T GAG TTCTACGGTCT C AG C GAGAAT GAT GAG T GGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGA AATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAA TGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGTACTAATTATGCTCTCTTGAAA TTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCAGCTGCTCCTCCCAACTGCTCT GCTACTTCTAGTTTCAGCTGGCATGCGGACTGAAGATCTCCCAAAGGCTGTGGTGTTCCTGG AGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACTCTGAAGTGCCAGGGAGCCTAC TCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAGAGTACAGGTGCCAGACAAACC TCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATCGGCTGGCTGTTGCTCCAGGCC CCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAGGTGTCACAGCTGGAAGAACAC TGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCAGGAAGTATTTTCATCATAATT CTGACTTCTACATTCCAAAAGCCACACTCAAAGACAGCGGCTCCTACTTCTGCAGGGGGCTT gTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCACCATCACTCAAGGTTTGGCAGT GTCAACCATCTCATCATTCTTTCCACCTGGGTACCAAgtctctttctgcttggtgatggtac tcctttttgcagtggacacaggactatatttctctgtgaagacaaacattcgaagctcaaca agagactggaaggaccataaatttaaatggagaaaggaccctcaagacaaaTAA ( SEQ IDNO : 172 )TCR / FcR #3 open reading frame (SEQ ID NO: 173)ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATTTCAGCCTGCTGAAACAGGCTG GCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTGCTGTGCTATATGGGCTTCTAC TTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGT ATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACC GAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACT GACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGG CGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAAT GTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAA GGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCC GCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCA AAAC C C AAGAAAT GAG T T C AGAT G T C AAG T T GAG TTCTACGGTCT GAG C GAGAAT GAT GAG T GGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGA AATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAA TGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGTACTAATTATGCTCTCTTGAAA TTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCAGCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGACTGAAGATCTCCCAAAGGCTGTGGTGTTCCTGG AGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACTCTGAAGTGCCAGGGAGCCTAC TCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAGCCTCATCTCAAGCCAGGCCTC GAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAGAGTACAGGTGCCAGACAAACC TCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATCGGCTGGCTGTTGCTCCAGGCC CCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAGGTGTCACAGCTGGAAGAACAC TGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCAGGAAGTATTTTCATCATAATT CTGACTTCTACATTCCAAAAGCCACACTCAAAGACAGCGGCTCCTACTTCTGCAGGGGGCTT gTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCACCGTCCAAGTGCCCAGCATGGG CAGCTCTTCACCAATGGGGgtctctttctgcttggtgatggtactcctttttgcagtggaca caggactatatttctctgtgaagacaaacattcgaagctcaacaagagactggaaggaccat aaatttaaatggagaaaggaccctcaagacaaaTAA ( SEQ ID NO : 173 )TCR / FcR #4 open reading frame (SEQ ID NO: 174)ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGAT GACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATTTCAGCCTGCTGAAACAGGCTG GCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTGCTGTGCTATATGGGCTTCTAC TTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGG CACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGT ATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACC GAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACT GACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGG CGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAAT GTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAA GGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGG TTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCC GCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCA AAAC C C AAGAAAT GAG T T C AGAT G T C AAG T T GAG TTCTACGGTCT GAG C GAGAAT GAT GAG T GGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCG GATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGA AATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAA TGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGTACTAATTATGCTCTCTTGAAA TTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCAGCTGCTCCTCCCAACTGCTCT GCTACTTCTAGTTTCAGCTGGCATGCGGACTGAAGATCTCCCAAAGGCTGTGGTGTTCCTGG AGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACTCTGAAGTGCCAGGGAGCCTAC TCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAGCCTCATCTCAAGCCAGGCCTC GAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAGAGTACAGGTGCCAGACAAACC TCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAGGTGTCACAGCTGGAAGAACAC TGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCAGGAAGTATTTTCATCATAATT CTGACTTCTACATTCCAAAAGCCACACTCAAAGACAGCGGCTCCTACTTCTGCAGGGGGCTT gTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCACCGTCCAAGTGCCCAGCATGGG CAGCTCTTCACCAATGGGGCTCTGTTACCTTCTTGACGGTATTCTTTTTATTTACGGCGTCA TCCTCACTGCCCTCTTTTTGAGGGTCAAGTTTTCAAGATCCGCCGACGCACCTGCTTACCAG CAAGGACAAAATCAACTTTATAACGAACTCAACCTTGGTCGGCGGGAAGAATATGATGTACT CGACAAAAGAAGAGGCCGCGACCCGGAGATGGGTGGCAAACCACAGCGGAGGAAAAACCCCC AGGAAGGTCTCTATAACGAATTGCAGAAAGATAAAATGGCCGAGGCTTATTCAGAGATCGGC ATGAAAGGAGAGCGACGACGAGGAAAGGGGCATGATGGTCTGTATCAAGGGCTCAGCACGGC AACAAAGGACACATATGATGCCTTGCATATGCAGGCGCTTCCGCCGCGCTAA ( SEQ ID NO : 174 )TCR / FcR #5 open reading frame (SEQ ID NO: 175)ATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCTGTACTTCTACAGAGGCAACGG CAAGAACCAGGTGGAACAGAGCCCTCAGAGCCTGATCATCCTGGAAGGCAAGAACTGCACCC TGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGGTGGTACAAGCAGGATACAGGC AGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTA CACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGCACATCACAGCCAGCCAGCTGA GCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGCAGCACCCTGGGCAGACTGTAC TTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCAGAACCCTGATCCTGCCGTGTA CCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCCTGTTCACCGACTTCGACAGCC AGACCAACGTGTCCCAGAGCAAGGACAGCGACGTGTACATCACCGACAAGtgcGTGCTGGAC ATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTGGTCCAACAAGAGCGATTTCGC CTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACACATTCTTCCCAAGTCCTGAGA GCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAG AACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATTTCAGCCTGCTGAAACAGGCTG GCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTGCTGTGCTATATGGGCTTCTAC TTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGACACCTAGATACCTGGTCATCGG CACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGGGCCACGACAAGATGTACTGGT ATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTACAGCTACGGCGTGAACAGCACC GAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAATCCGGACCGAGCACTTCCCACT GACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACCTGTGTGCCACTGGACAGGGGG CGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACAGTGCTCGAGGACCTCAAGAAT GTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGCCGAGATCTCTCATACACAAAA GGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACCACGTCGAGCTTTCCTGGTGGG TTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCTCAGCCATTGAAGGAACAGCCC GCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCGCGTGTCTGCGACATTCTGGCA AAAC C C AAGAAAT GAG T T C AGAT G T C AAG T T GAG TTCTACGGTCT GAG C GAGAAT GAT GAG TGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCTGCCGAGGCCTGGGGGAGGGCG GATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATTGAGCGCGACAATTCTTTACGA AATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGTCCGCTTTGGTTCTTATGGCAA TGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGTACTAATTATGCTCTCTTGAAA TTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCAGCTGCTCCTCCCAACTGCTCT GCTACTTCTAGTTTCAGCTGGCATGCGGACTGAAGATCTCCCAAAGGCTGTGGTGTTCCTGG AGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACTCTGAAGTGCCAGGGAGCCTAC TCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAGCCTCATCTCAAGCCAGGCCTC GAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAGAGTACAGGTGCCAGACAAACC TCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAGGTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCAGGAAGTATTTTCATCATAATTCTGACTTCTACATTCCAAAAGCCACACTCAAAGACAGCGGCTCCTACTTCTGCAGGGGGCTT gTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCACCGTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGGgtctctttctgcttggtgatggtactcctttttgcagtggaca caggactatatttctctgtgaagacaTAA ( SEQ ID NO : 175 ) iTCR3 transgenic vector (SEQ ID NO: 176)AATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCAT GGAAAAATACATAAC T GAGAATAGAAAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAAC AGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCT CAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCAT CAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAA TCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCAC AACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCA ATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCT GAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGAC CCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTC TGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAAC TAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCC TGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCC GATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGA CGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCC GCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTG TATTTGTCTGAAAATATGGGCCCGGGCTAGCCTGTTACCACTCCCTTAAGTTTGACCTTAGG TCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTT GGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACC TTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACA CCCAGACCAGGTGGGGTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGG TCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCC CTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCT AGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCC CTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTC TACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCG ACCGGTGGTACCTCACCCTTACCGAGTCGGCGACACAGTGTGGGTCCGCCGACACCAGACTA AGAACCTAGAACCTCGCTGGAAAGGACCTTACACAGTCCTGCTGACCACCCCCACCGCCCTC AAAGTAGACGGCATCGCAGCTTGGATACACGCCGCCCACGTGAAGGCTGCCGACCCCGGGGG T G GAG C AT C C T C T AGAC TGC CATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCT GTACTTCTACAGAGGCAACGGCAAGAAC GAG G T G GAAC AGAG C C C T C AGAG CCTGATCATCC TGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGG TGGTACAAGCAGGATACAGGCAGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAA CACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGC ACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGC AGCACCCTGGGCAGACTGTACTTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCA GAACCCTGATCCTGCCGTGTACCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCC T G T T GAG C GAG T T C GAG AG C C AGAC C AAC G T G T C C C AGAG C AAG GAC AG C GAC G T G TAG AT C ACCGACAAGtgcGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTG GTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACA CATTCTTCCCAAGTCCTGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACAGACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGT GGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATT TCAGCCTGCTGAAACAGGCTGGCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTG CTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGAC ACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGG GCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTAC AGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAAT CCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACC TGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACA GTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGC CGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACC ACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCT CAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCG CGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACG GTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCT GCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATT GAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGT CCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGCTAACAATTGCGCGTC ATCATCGATCCGGATTAGTCCAATTTGTTAAAGACAGGATATCAGTGGTCCAGGCTCTAGTT T T GAG T C AAC AAT AT GAG GAG C T GAAG C C T AT AGAG TAG GAG C C AT AGAT AAAAT AAAAGAT TTTATTTAGTCTCCAGAAAAAGGGGGGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAG C T TAAGTAACGCCAT T T T GCAAGGCAT GGAAAAAT AGAT AAC T GAGAATAGAGAAGT T GAGA TCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAG TTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATAT CTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCC AGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATG ACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTG CTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGAC TGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCA ( SEQ ID NO : 17 6 )TCR / FcR #1 transgenic vector (SEQ ID NO: 177)AATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCAT GGAAAAAT AGAT AAC T GAGAATAGAAAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAAC AGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCT CAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCAT CAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAA TCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCAC AACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCA ATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCT GAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGAC CCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTC TGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAAC TAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCC TGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCC GATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGA CGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCC GCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATATGGGCCCGGGCTAGCCTGTTACCACTCCCTTAAGTTTGACCTTAGG TCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTT GGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACC TTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACA CCCAGACCAGGTGGGGTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGG TCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCC CTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCT AGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCC CTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTC TACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCG ACCGGTGGTACCTCACCCTTACCGAGTCGGCGACACAGTGTGGGTCCGCCGACACCAGACTA AGAACCTAGAACCTCGCTGGAAAGGACCTTACACAGTCCTGCTGACCACCCCCACCGCCCTC AAAGTAGACGGCATCGCAGCTTGGATACACGCCGCCCACGTGAAGGCTGCCGACCCCGGGGG T G GAG C AT C C T C T AGAC TGC CATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCT GTACTTCTACAGAGGCAACGGCAAGAAC GAG G T G GAAC AGAG C C C T C AGAG CCTGATCATCC TGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGG TGGTACAAGCAGGATACAGGCAGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAA CACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGC ACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGC AGCACCCTGGGCAGACTGTACTTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCA GAACCCTGATCCTGCCGTGTACCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCC T G T T GAG C GAG T T C GAG AG C C AGAC C AAC G T G T C C C AGAG C AAG GAC AG C GAC G T G TAG AT C ACCGACAAGtgcGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTG GTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACA CATTCTTCCCAAGTCCTGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACA GACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGT GGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATT TCAGCCTGCTGAAACAGGCTGGCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTG CTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGAC ACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGG GCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTAC AGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAAT CCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACC TGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACA GTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGC CGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACC ACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCT CAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCG CGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACG GTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCT GCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATT GAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGT CCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCA GCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GAAGAT C T CC CAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACTCTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAGAGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATCGGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAGGTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCAGGAAGTAT T T T CAT CATAAT T C T GAG T T C TACAT T CCAAAAGCCACAC T CAAAGACAGCGGC TCCTACTTCTGCAGGGGGCTTTTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCAC CATCACTCAAGGTTTGGCAGTGTCAACCATCTCATCATTCTTTCCACCTGGGTACCAAgtct ctttctgcttggtgatggtactcctttttgcagtggacacaggactatatttctctgtgaag acaaacattcgaagctcaacaagagactggaaggaccataaatttaaatggagaaaggaccc tcaagacaaaTAACAATTGCGCGTCATCATCGATCCGGATTAGTCCAATTTGTTAAAGACAG GATATCAGTGGTCCAGGCTCTAGTTTTGACTCAACAATATCACCAGCTGAAGCCTATAGAGT ACGAGCCATAGATAAAATAAAAGAT T T TAT T TAGT C T CCAGAAAAAGGGGGGAAT GAAAGAC CCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATAC AT AAC T GAGAATAGAGAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAAT AT GGGC CAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACA GCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAG AACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTC CAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCT TCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCAC TCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTC TTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGA CTACCCGTCAGCGGGGGTCTTTCA (SEQ ID NO: 177)TCR / FcR #2 transgenic vector (SEQ ID NO: 178)AATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCAT GGAAAAATACATAAC T GAGAATAGAAAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAAC AGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCT CAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCAT CAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAA TCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCAC AACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCA ATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCT GAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGAC CCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTC TGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAAC TAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCC TGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCC GATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGA CGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCC GCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTG TATTTGTCTGAAAATATGGGCCCGGGCTAGCCTGTTACCACTCCCTTAAGTTTGACCTTAGG TCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTT GGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACC TTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACA CCCAGACCAGGTGGGGTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGG TCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCC CTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCT AGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCC CTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTC TACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCG ACCGGTGGTACCTCACCCTTACCGAGTCGGCGACACAGTGTGGGTCCGCCGACACCAGACTA AGAACCTAGAACCTCGCTGGAAAGGACCTTACACAGTCCTGCTGACCACCCCCACCGCCCTC AAAGTAGACGGCATCGCAGCTTGGATACACGCCGCCCACGTGAAGGCTGCCGACCCCGGGGGT G GAG C AT C C T C T AGAC TGC CATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCT GTACTTCTACAGAGGCAACGGCAAGAAC GAG G T G GAAC AGAG C C C T C AGAG CCTGATCATCC TGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGG TGGTACAAGCAGGATACAGGCAGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAA CACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGC ACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGC AGCACCCTGGGCAGACTGTACTTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCA GAACCCTGATCCTGCCGTGTACCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCC T G T T GAG C GAG T T C GAG AG C C AGAC C AAC G T G T C C C AGAG C AAG GAC AG C GAC G T G TAG AT C ACCGACAAGtgcGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTG GTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACA CATTCTTCCCAAGTCCTGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACA GACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGT GGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATT TCAGCCTGCTGAAACAGGCTGGCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTG CTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGAC ACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGG GCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTAC AGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAAT CCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACC TGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACA GTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGC CGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACC ACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCT CAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCG CGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACG GTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCT GCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATT GAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGT CCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGT ACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCA GCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GAAGAT C T CC CAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACT CTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAG CCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAG AGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATC GGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAG GTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCA GGAAGTAT T T T CAT CATAAT T C T GAC T T C TACAT T CCAAAAGCCACAC T CAAAGACAGCGGC TCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCAC CATCACTCAAGGTTTGGCAGTGTCAACCATCTCATCATTCTTTCCACCTGGGTACCAAgtct ctttctgcttggtgatggtactcctttttgcagtggacacaggactatatttctctgtgaag acaaacattcgaagctcaacaagagactggaaggaccataaatttaaatggagaaaggaccc tcaagacaaaTAACAATTGCGCGTCATCATCGATCCGGATTAGTCCAATTTGTTAAAGACAG GATATCAGTGGTCCAGGCTCTAGTTTTGACTCAACAATATCACCAGCTGAAGCCTATAGAGT ACGAGCCATAGATAAAATAAAAGAT T T TAT T TAGT C T CCAGAAAAAGGGGGGAAT GAAAGAC CCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATAC ATAAC T GAGAATAGAGAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATAT GGGC CAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACA GCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAG AACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGA CTACCCGTCAGCGGGGGTCTTTCA ( SEQ ID NO : 178 )TCR / FcR #3 transgenic vector (SEQ ID NO: 179)AATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCAT GGAAAAATACATAAC T GAGAATAGAAAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAAC AGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCT CAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCAT CAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAA TCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCAC AACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCA ATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCT GAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGAC CCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTC TGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAAC TAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCC TGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCC GATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGA CGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCC GCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTG TATTTGTCTGAAAATATGGGCCCGGGCTAGCCTGTTACCACTCCCTTAAGTTTGACCTTAGG TCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTT GGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACC TTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACA CCCAGACCAGGTGGGGTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGG TCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCC CTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCT AGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCC CTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTC TACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCG ACCGGTGGTACCTCACCCTTACCGAGTCGGCGACACAGTGTGGGTCCGCCGACACCAGACTA AGAACCTAGAACCTCGCTGGAAAGGACCTTACACAGTCCTGCTGACCACCCCCACCGCCCTC AAAGTAGACGGCATCGCAGCTTGGATACACGCCGCCCACGTGAAGGCTGCCGACCCCGGGGG T G GAG C AT C C T C T AGAC TGC CATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCT GTACTTCTACAGAGGCAACGGCAAGAAC GAG G T G GAAC AGAG C C C T C AGAG CCTGATCATCC TGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGG TGGTACAAGCAGGATACAGGCAGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAA CACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGC ACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGC AGCACCCTGGGCAGACTGTACTTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCA GAACCCTGATCCTGCCGTGTACCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCC T G T T GAG C GAG T T C GAG AG C C AGAC C AAC G T G T C C C AGAG C AAG GAC AG C GAC G T G TAG AT C ACCGACAAGtgcGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTG GTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACA CATTCTTCCCAAGTCCTGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACA GACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGTGGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATT TCAGCCTGCTGAAACAGGCTGGCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTG CTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGAC ACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGG GCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTAC AGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAAT CCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACC TGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACA GTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGC CGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACC ACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCT CAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCG CGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACG GTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCT GCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATT GAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGT CCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGT ACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCA GCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GAAGAT C T CC CAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACT CTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAG CCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAG AGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATC GGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAG GTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCA GGAAGTAT T T T CAT CATAAT T C T GAG T T C TACAT T CCAAAAGCCACAC T CAAAGACAGCGGC TCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCAC CGTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGGgtctctttctgcttggtgatgg tactcctttttgcagtggacacaggactatatttctctgtgaagacaaacattcgaagctca acaagagactggaaggaccataaatttaaatggagaaaggaccctcaagacaaaTAACAATT GCGCGTCATCATCGATCCGGATTAGTCCAATTTGTTAAAGACAGGATATCAGTGGTCCAGGC TCTAGTTTT GAG T C AAC AAT AT GAG GAG C T GAAG C C T AT AGAG TAG GAG C C AT AGAT AAAAT AAAAGATTTTATTTAGTCTCCAGAAAAAGGGGGGAATGAAAGACCCCACCTGTAGGTTTGGC AAGC TAGC T TAAGTAACGCCAT T T T GCAAGGCAT GGAAAAAT AGAT AAC T GAGAATAGAGAA GTTCAGATCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGG TAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAAC AGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGAT GCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACC TGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGC GCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCC GATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACT TGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGT CTTTCA ( SEQ ID NO : 179 )TCR / FcR #4 transgenic vector (SEQ ID NO: 180)AATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCAT GGAAAAAT ACAT AAC T GAGAATAGAAAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAAC AGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCT CAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAA TCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCAC AACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCA ATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCT GAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGAC CCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTC TGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAAC TAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCC TGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCC GATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGA CGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCC GCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTG TATTTGTCTGAAAATATGGGCCCGGGCTAGCCTGTTACCACTCCCTTAAGTTTGACCTTAGG TCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTT GGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACA CCCAGACCAGGTGGGGTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGG TCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCC CTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCT AGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCC CTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTC TACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCG ACCGGTGGTACCTCACCCTTACCGAGTCGGCGACACAGTGTGGGTCCGCCGACACCAGACTAAGAACCTAGAACCTCGCTGGAAAGGACCTTACACAGTCCTGCTGACCACCCCCACCGCCCTC AAAGTAGACGGCATCGCAGCTTGGATACACGCCGCCCACGTGAAGGCTGCCGACCCCGGGGG T G GAG C AT C C T C T AGAC TGC CATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCT GTACTTCTACAGAGGCAACGGCAAGAAC C AG G T G GAAC AGAG C C C T C AGAG CCTGATCATCC TGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGG TGGTACAAGCAGGATACAGGCAGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAACACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGC ACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGC AGCACCCTGGGCAGACTGTACTTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCA GAACCCTGATCCTGCCGTGTACCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCC T G T T GAG C GAG T T C GAG AG C C AGAC C AAC G T G T C C C AGAG C AAG GAG AG C GAG G T G TAG AT C ACCGACAAGtgcGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTG GTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACA CATTCTTCCCAAGTCCTGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACA GACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGT GGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATT TCAGCCTGCTGAAACAGGCTGGCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTG CTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGAC ACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGG GCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTAC AGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAAT CCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACC TGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACA GTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGC CGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACC ACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCT CAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCG CGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACGGTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCT GCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATT GAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGT CCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGT ACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCA GCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GAAGAT C T CC CAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACT CTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAG CCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAG AGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATC GGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAG GTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCA GGAAGTAT T T T CAT CATAAT T C T GAG T T C TACAT T CCAAAAGCCACAC T CAAAGACAGCGGC TCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCAC CGTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGGCTCTGTTACCTTCTTGACGGTA TTCTTTTTATTTACGGCGTCATCCTCACTGCCCTCTTTTTGAGGGTCAAGTTTTCAAGATCC GCCGACGCACCTGCTTACCAGCAAGGACAAAATCAACTTTATAACGAACTCAACCTTGGTCG GCGGGAAGAATATGATGTACTCGACAAAAGAAGAGGCCGCGACCCGGAGATGGGTGGCAAAC CACAGCGGAGGAAAAACCCCCAGGAAGGTCTCTATAACGAATTGCAGAAAGATAAAATGGCC GAGGCTTATTCAGAGATCGGCATGAAAGGAGAGCGACGACGAGGAAAGGGGCATGATGGTCT GTATCAAGGGCTCAGCACGGCAACAAAGGACACATATGATGCCTTGCATATGCAGGCGCTTC CGCCGCGCTAATAACAATTGCGCGTCATCATCGATCCGGATTAGTCCAATTTGTTAAAGACA GGATATCAGTGGTCCAGGCTCTAGTTTTGACTCAACAATATCACCAGCTGAAGCCTATAGAG TACGAGCCATAGATAAAATAAAAGAT T T TAT T TAGT C T CCAGAAAAAGGGGGGAAT GAAAGA CCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATA CATAAC T GAGAATAGAGAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATAT GGG CCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAAC AGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAA GAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTT CCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGC TTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCA CTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCT CTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTG ACTACCCGTCAGCGGGGGTCTTTCA ( SEQ ID NO : 180 )TCR / FcR #5 transgenic vector (SEQ ID NO: 181])AATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCAT GGAAAAATACATAAC T GAGAATAGAAAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAAC AGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCT CAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCAT CAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAA TCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCAC AACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCA ATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCT GAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGAC CCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTC TGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAAC TAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCC TGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCCGATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGA CGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCC GCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTG TATTTGTCTGAAAATATGGGCCCGGGCTAGCCTGTTACCACTCCCTTAAGTTTGACCTTAGG TCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTT GGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACC TTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACA CCCAGACCAGGTGGGGTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGG TCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCC CTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCT AGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCC CTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTC TACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCG ACCGGTGGTACCTCACCCTTACCGAGTCGGCGACACAGTGTGGGTCCGCCGACACCAGACTA AGAACCTAGAACCTCGCTGGAAAGGACCTTACACAGTCCTGCTGACCACCCCCACCGCCCTC AAAGTAGACGGCATCGCAGCTTGGATACACGCCGCCCACGTGAAGGCTGCCGACCCCGGGGG T G GAG C AT C C T C T AGAC TGC CATGAAGAAGCACCTGACCACCTTTCTGGTCATCCTGTGGCT GTACTTCTACAGAGGCAACGGCAAGAAC GAG G T G GAAC AGAG C C C T C AGAG CCTGATCATCC TGGAAGGCAAGAACTGCACCCTGCAGTGCAACTACACCGTGTCTCCCTTCAGCAACCTGCGG TGGTACAAGCAGGATACAGGCAGAGGCCCTGTGTCTCTGACCATCATGACCTTCAGCGAGAA CACCAAGAGCAACGGCCGGTACACCGCCACACTGGATGCCGATACAAAGCAGAGCAGCCTGC ACATCACAGCCAGCCAGCTGAGCGATAGCGCCAGCTACATCTGCGTGGTGTCCGATAGAGGC AGCACCCTGGGCAGACTGTACTTTGGCAGAGGCACCCAGCTGACCGTGTGGCCCGATATTCA GAACCCTGATCCTGCCGTGTACCAGCTGAGAGACAGCAAGTCCAGCGACAAGAGCGTGTGCC T G T T GAG C GAG T T C GAG AG C C AGAC C AAC G T G T C C C AGAG C AAG GAC AG C GAC G T G TAG AT C ACCGACAAGtgcGTGCTGGACATGCGGAGCATGGACTTCAAGAGCAATAGCGCCGTGGCCTG GTCCAACAAGAGCGATTTCGCCTGCGCCAACGCCTTCAACAACAGCATTATCCCCGAGGACA CATTCTTCCCAAGTCCTGAGAGCAGCTGCGACGTGAAGCTGGTGGAAAAGAGCTTCGAGACA GACACCAACCTGAACTTCCAGAACCTGAGCGTGATCGGCTTCCGGATCCTGCTGCTGAAAGT GGCCGGCTTCAACCTGCTGATGACCCTGAGACTGTGGTCTAGCGGAAGCGGCGCCACCAATT TCAGCCTGCTGAAACAGGCTGGCGACGTGGAAGAGAACCCCGGACCTATGACCATCCGGCTG CTGTGCTATATGGGCTTCTACTTCCTCGGAGCCGGCCTGATGGAAGCCGACATCTACCAGAC ACCTAGATACCTGGTCATCGGCACCGGCAAAAAGATCACCCTGGAATGCTCCCAGACCATGG GCCACGACAAGATGTACTGGTATCAGCAGGACCCCGGCATGGAACTGCATCTGATCCACTAC AGCTACGGCGTGAACAGCACCGAGAAGGGCGATCTGTCTAGCGAGAGCACCGTGTCCAGAAT CCGGACCGAGCACTTCCCACTGACACTGGAAAGCGCCAGACCTAGCCACACCAGCCAGTACC TGTGTGCCACTGGACAGGGGGCGCAAGATACGCAGTATTTTGGCCCAGGCACCCGGCTGACA GTGCTCGAGGACCTCAAGAATGTGTTTCCGCCCGAAGTCGCGGTTTTTGAACCATCAGAAGC CGAGATCTCTCATACACAAAAGGCGACGCTCGTATGCCTcGCGACGGGATTTTATCCGGACC ACGTCGAGCTTTCCTGGTGGGTTAAcGGAAAGGAGGTGCATTCCGGAGTTTGCACGGACCCT CAGCCATTGAAGGAACAGCCCGCACTGAACGACAGTAGGTATTGCCTTTCATCTCGCCTGCG CGTGTCTGCGACATTCTGGCAAAACCCAAGAAATCACTTCAGATGTCAAGTTCAGTTCTACG GTCTCAGCGAGAATGATGAGTGGACACAAGATAGGGCTAAACCCGTGACTCAAATAGTCTCT GCCGAGGCCTGGGGGAGGGCGGATTGCGGCTTCACATCAGAATCATACCAACAAGGAGTATT GAGCGCGACAATTCTTTACGAAATTCTGCTTGGGAAAGCGACTCTGTACGCGGTGCTCGTGT CCGCTTTGGTTCTTATGGCAATGGTTAAACGAAAGGATAGTAGGGGCGGtAGtGGaCAGTGTACTAATTATGCTCTCTTGAAATTGGCTGGAGATGTTGAGAGCAATCCCGGGCCCATGTGGCA GCTGCTCCTCCCAACTGCTCTGCTACTTCTAGTTTCAGCTGGCATGCGGAC T GAAGAT C T CC CAAAGGCTGTGGTGTTCCTGGAGCCTCAATGGTACAGGGTGCTCGAGAAGGACAGTGTGACTCTGAAGTGCCAGGGAGCCTACTCCCCTGAGGACAATTCCACACAGTGGTTTCACAATGAGAGCCTCATCTCAAGCCAGGCCTCGAGCTACTTCATTGACGCTGCCACAGTCGACGACAGTGGAGAGTACAGGTGCCAGACAAACCTCTCCACCCTCAGTGACCCGGTGCAGCTAGAAGTCCATATC GGCTGGCTGTTGCTCCAGGCCCCTCGGTGGGTGTTCAAGGAGGAAGACCCTATTCACCTGAG GTGTCACAGCTGGAAGAACACTGCTCTGCATAAGGTCACATATTTACAGAATGGCAAAGGCA GGAAGTAT T T T CAT CATAAT T C T GAG T T C TACAT T CCAAAAGCCACAC T CAAAGACAGCGGC TCCTACTTCTGCAGGGGGCTTgTTGGGAGTAAAAATGTGTCTTCAGAGACTGTGAACATCAC CGTCCAAGTGCCCAGCATGGGCAGCTCTTCACCAATGGGGgtctctttctgcttggtgatgg tactcctttttgcagtggacacaggactatatttctctgtgaagacaTAACAATTGCGCGTC ATCATCGATCCGGATTAGTCCAATTTGTTAAAGACAGGATATCAGTGGTCCAGGCTCTAGTT T T GAG T C AAC AAT AT GAG GAG C T GAAG C C T AT AGAG TAG GAG C C AT AGAT AAAAT AAAAGAT TTTATTTAGTCTCCAGAAAAAGGGGGGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAG C T TAAGTAACGCCAT T T T GCAAGGCAT GGAAAAAT AGAT AAC T GAGAATAGAGAAGT T CAGA TCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAG TTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATAT CTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCC AGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATG ACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTG CTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGAC TGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTC TCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCA ( SEQ ID NO : 181 )D. NK Cells

[0242] The NK cells that are modified to express the TCR / CD3 receptor complex may be obtained from any suitable source, including fresh or frozen sources. In certain embodiments, NK cells are not NK cells obtained from iPSC differentiation. In certain embodiments, NK cells are not from an NK cell line (e.g., NK-92). In certain embodiments, NK cells are derived from human peripheral blood mononuclear cells (PBMC), unstimulated leukapheresis products (PBSC), NK cell lines (e.g., NK-92), human embryonic stem cells (hESCs), induced pluripotent stem cells (iPSCs), bone marrow, or umbilical cord blood by methods well known in the art. Specifically, the NK cells may be isolated from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, NK cell lines, or a mixture thereof. In particular embodiments, the NK cells are isolated from pooled CB. The CB may be pooled from 2, 3, 4, 5, 6, 7, 8, 9, 10, or more units. The NK cells may be autologous or allogeneic with respect to a recipient individual. The isolated NK cells may or may not be haplotype matched for the subject to be administered the cell therapy. NK cells can be detected by specific surface markers, such as CD 16 and CD56 in humans, for example. In some cases, the source of the NK cells is cord blood and the NK cells may be in the cord blood in a heterogeneous mixture of cells and may be depleted of certain cells expressing CD3. In other methods, umbilical CB is used to derive NK cells by the isolation of CD34+ cells.

[0243] The NK cells may be pre-activated with one or more inflammatory cytokines, and they may be expanded or non-expanded. In some cases, the NK cells are pre-activated eitherprior to modification (e.g., engineering) or after modification. In specific embodiments, preactivation of the NK cells may comprise culturing the isolated NK cells in the presence of one or more cytokines. The NK cells may be stimulated with IL-2, or other cytokines that bind the common gamma-chain (e.g., IL-7, IL-12, IL-15, IL-18, IL-21, and others). In particular embodiments, the pre-activation cytokines may be selected from the group consisting of IL- 12, IL-15, IL-18, and a combination thereof. One or more additional cytokines may be used for the pre-activation step. The pre-activation may be for a short period of time such as 5-72 hours, such as 10-50 hours, particularly 10-20 hours, such as 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours, specifically about 16 hours. The pre-activation culture may comprise IL-12 at a concentration of 0.1-150 ng / mL, such as 0.5-50 ng / mL, particularly 1-20 ng / mL, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ng / mL, specifically about 10 ng / mL. The pre-activation culture may comprise IL- 18 and / or IL- 15 at a concentration of 10-100 ng / mL, such as 40-60 ng / mL, particularly 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 ng / mL, specifically about 50 ng / mL.

[0244] In some cases, the NK cells are expanded either prior to modification to express constructs described herein (e.g., uTNKl 5 and / or TCR / FcR constructs). Pre-activated NK cells may be expanded in the presence of artificial antigen presenting cells (aAPCs) and / or feeders / fragments or NK activating beads. The pre-activated NK cells may be washed prior to expansion, such as 2, 3, 4, or 5 times, specifically 3 times. The aAPCs may be engineered to express CD137 ligand and / or a membrane-bound cytokine. The membrane-bound cytokine may be membrane-bound IL-21 (mIL-21) or membrane-bound IL- 15 (mIL-15). In particular embodiments, the aAPCs are engineered to express CD137 ligand and mIL-2L The aAPCs may be derived from cancer cells, such as leukemia cells. The aAPCs may not express endogenous HLA class I, II, or CD Id molecules. They may express ICAM-1 (CD54) and LFA- 3 (CD58). In particular, the aAPCs may be K562 cells, such as K562 cells engineered to express CD137 ligand and mIL-21. The aAPCs may be irradiated. In some embodiments, fragments of APC can be used to expand the NK cells. The engineering may be by any method known in the art, such as retroviral transduction. Retroviral transduction may be at least, at most, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days following NK co-culturing with an antigen presenting cell. In some embodiments, retroviral transduction comprises cotransduction of more than one construct. In some embodiments, retroviral transduction occurs after or at about 5 days of co-culturing with an antigen presenting cell. In some embodiments, co-culturing with an antigen presenting cell continues following transduction of an NK cell. The expansion may be for about 2-30 days, such as 3-20 days, particularly 12-16 days, such as12, 13, 14, 15, 16, 17, 18, or 19 days, specifically about 14 days. The pre-activated NK cells and aAPCs may be present at a ratio of about 3: 1-1 :3, such as 2: 1, 1 : 1, 1 :2, specifically about 1 :2. The expansion culture may further comprise cytokines to promote expansion, such as IL- 2. The IL-2 may be present at a concentration of about 10-500 U / mL, such as 100-300 U / mL, particularly about 200 U / mL. The IL-2 may be replenished in the expansion culture, such as every 2-3 days. The aAPCs may be added to the culture at least a second time, such as at about 7 days of expansion.

[0245] In particular embodiments, the NK cells are transfected or transduced with one or more membrane bound cytokines, including IL-21, IL-12, IL-18, IL-23, IL-7, or IL-15, either secreted by NK cells or tethered to the NK cell membrane. In such cases, the membrane bound cytokine may be tethered to the NK cell membrane with a particular transmembrane domain, such as the transmembrane domain of CD8, CD28, CD27, B7H3, IgGl, IgG4, CD4, DAP10, DAP 12, for example.

[0246] Following preparation, the modified NK cells may be immediately infused (including with an effective amount of one or more bispecific or multi-specific antibodies, or the NK cells may be stored, such as by cryopreservation. In some cases, when the NK cells are source from cryopreservation, the NK cells were deactivated pre-cryopreservation using a deactivating agent (e.g., a kinase inhibitor, e.g., Dasatinib, nilotinib, rapamycin, etc.). In certain aspects, the cells may be propagated for days, weeks, or months ex vivo as a bulk population within about 1, 2, 3, 4, or 5 days.E. Loading of NK Cells

[0247] In particular embodiments, the NK cells are loaded with antibodies prior to use. The NK cells may be loaded in any specific manner, including in culture (e.g., incubation) or immediately before infusion, for example, to produce a complex of NK cells with the antibodies. The culture (e.g., incubation) conditions are suitable enough to allow for an effective amount of antibody to bind to the surface of the NK cells. In the case of use of monospecific antibodies, the Fc region of the monospecific antibody binds the NK cell while the antigen binding domain of the monospecific antibody is free to bind its target antigen. In certain embodiments of cases of use of multispecific antibodies, one or more antigen binding domains of the antibody can bind the surface of the NK cells, such as through an antigen on the surface of the NK cells (for example but not limited to, NKp30, NKp44, NKp46, CD16, CD32, CD64, KIRs, and the like), and the other antigen binding domain is free to bind its target antigen. In certain embodiments of cases of use of multispecific antibodies, one or more antigenbinding domains of the antibody can bind one or more target antigens. In certain embodiments of cases of use of multispecific antibodies, the Fc region of the antibody binds the NK cell while the antigen binding domains of the antibody are free to bind target antigens. In certain embodiments of cases of use of multispecific antibodies, the Fc region of the antibody binds the NK cell through a transgenic construct (heterologous construct) encoded polypeptide comprising an Fc Receptor extracellular Fc binding domain (e.g., an Fc binding domain).

[0248] The culture conditions by which the NK cells become loaded may or may not be of a particular type having one or more specific parameters. In particular embodiments, the loading of the NK cells occurs in culture at a specific temperature, such as 37 °C, although in alternative embodiments the temperature is 36 °C or 38 °C, or lower or higher. The duration of the loading step may be for any suitable amount of time, such as in a range of one minute to 24 hours or longer. For example, the range may be in the range of 1 min to 24 hrs, 1 min to 18 hrs, 1 min to 12 hours, 1 min to 6 hrs, 1 min to 1 hr, 30 min to 24 hrs, 30 min to 18 hrs, 30 min to 12 hrs, 30 min to 6 hrs, 30 min to 1 hr, 1-24 hrs, 1-18 hrs, 1-12 hrs, 1-6 hrs, 6-24 hrs, 6-18 hrs, 6-12 hrs, 12-24 hrs, 12-18 hrs, or 18-24 hrs. In some embodiments, the duration of the loading step may be greater than or equal to approximately 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, or 48 hours, or any range derivable therein. In specific embodiments, the cell culture media is basal media or complex media. In some cases, the culture comprises one or more reagents that were utilized during pre-activation and / or expansion steps, while in other cases the culture does not. In specific embodiments, the culture comprises one or more cytokines, including one or more of IL-12, IL-15, IL-2, and IL-18, for example. In some embodiments, the culture comprises APCs of any kind.

[0249] In some embodiments, loading of the NK cells is conducted such that antibodies remain bound to the NK cell surface following washing of the NK cells. In some embodiments, NK cells are loaded with antibodies under conditions that are suitable for retention of at least a detectable fraction of the antibody on the NK cell surface for at least or exactly 12 hours, 24 hours, 36 hours, 48 hours, 60 hours, 72 hours, 84 hours, 96 hours, 108 hours, 120 hours, 132 hours, 144 hours, 156 hours, 168 hours, 180 hours, 192 hours, 204 hours, 216 hours, 228 hours, 240 hours, 252 hours, 264 hours, 276 hours, 288 hours, 300 hours, 312 hours, or greater than 312 hours, or any range derivable therein. In some embodiments, NK cells are loaded with antibodies under conditions that are suitable for retention of the antibody on the NK cell surface for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15, or longer than 15 days, or any range derivable therein. In some embodiments, NK cells are loaded with antibodies under conditions that are suitable for retention of the antibody on the NK cell surface following cry opreservationand / or thawing. In some embodiments, confirmation of loading of an antibody to an NK cell surface can be determined using flow cytometry.

[0250] In some embodiments, NK cells are loaded with at least, exactly, or 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 pg / ml final concentration, or greater than 100 gg / ml final concentration, or any range derivable therein, of antibody. In some embodiments, NK cells are loaded with at least, exactly, or about 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, 860, 880, 900, 920, 940, 960, 980, or 1000 pg / ml final concentration, or greater than 1000 gg / ml final concentration, or any range derivable therein, of antibody.

[0251] In some embodiments, NK cells and antibodies are incubated such that at least or exactly 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%, or any range derivable therein, of NK cells are complexed with an antibody.

[0252] In some embodiments, NK cells and antibodies are incubated such that at least or exactly 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%, or any range derivable therein, of the antibody remains bound to the NK cells for at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days,7 days, 8 days, 9 days, 10 days, 11 days, or 12 days, or any range derivable therein, following incubation of the NK cells and antibody.

[0253] In certain embodiments, relative to the amount of antibodies complexed to engineered NK cells measured about 1 hour after incubation and washing, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or greater than 20%, or any range derivable therein, of the antibodies are bound to the NK cell surface at least 3 days after incubation and washing. In certain embodiments, relative to the amount of antibodies complexed to engineered NK cells measured about 1 hour after incubation and washing, at least about 9%, of the antibodies are bound to the NK cell surface at least 3 days after incubation and washing.

[0254] In certain embodiments, relative to the amount of antibodies complexed to engineered NK cells measured about 1 hour after incubation and washing, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or greater than 20%, or any range derivable therein, of the antibodies are bound to the NK cell surface at least 5 days after incubation and washing. In certain embodiments, relative to the amount of antibodies complexed to engineered NK cells measured about 1 hour after incubation and washing, at least about 6% of the antibodies are bound to the NK cell surface at least 5 days after incubation and washing.

[0255] In certain embodiments, relative to the amount of antibodies complexed to engineered NK cells measured about 1 hour after incubation and washing, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or greater than 20%, or any range derivable therein, of the antibodies are bound to the NK cell surface at least 7 days after incubation and washing. In certain embodiments, relative to the amount of antibodies complexed to the engineered NK cells measured about 1 hour after incubation and washing, at least about 4% of the antibodies are bound to the NK cell surface at least 7 days after incubation and washing.

[0256] In certain embodiments, relative to the amount of antibodies complexed to engineered NK cells measured about 1 hour after incubation and washing, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or greater than 20%, or any range derivable therein, of the antibodies are bound to the NK cell surface at least 12 days after incubation and washing. In certain embodiments, relative to the amount of antibodies complexed to the engineered NK cells measured about 1 hour after incubation and washing, at least about 3% of the antibodies are bound to the NK cell surface at least 12 days after incubation and washing.

[0257] In certain embodiments, a composition comprising engineered NK cells and antibodies is cryopreserved. In certain embodiments, a composition comprising engineered NK cells and antibodies is thawed from cryopreservation, and at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the engineered NK cells are complexed to an antibody.

[0258] In certain embodiments, relative to the amount of antibodies complexed to noncryopreserved engineered NK cells measured about 1 hour after incubation and washing, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or greater than 90%, or any range derivable therein, of the antibodies are bound to the NK cell surface after thawing from cryopreservation. In certain embodiments, relative to the amount of antibodies complexed to non-cryopreserved engineered NK cells measured about 1 hour after incubation and washing, at least about 30% of the antibodies are bound to the NK cell surface after thawing from cryopreservation. In certain embodiments, relative to the amount of antibodies complexed to non-cryopreserved engineered NK cells measured about 1 hour after incubation and washing, at least about 40% of the antibodies are bound to the NK cell surface after thawing from cryopreservation. In certain embodiments, relative to the amount of antibodies complexed to non-cryopreserved engineered NK cells measured about 1 hour after incubation and washing, at least about 50% of the antibodies are bound to the NK cell surface after thawing from cryopreservation. In certain embodiments, relative to the amount of antibodies complexed to non-cryopreserved engineered NK cells measured about 1 hour after incubation and washing, at least about 60% of the antibodies are bound to the NK cell surface after thawing from cry opreservation.

[0259] In some embodiments, incubation of the NK cells and antibody occurs in any suitable NK cell media known to one of skill in the art. In certain embodiments, incubation of the NK cells and antibody is performed in media comprising, consisting of, or consisting essentially of Click’ s / RPMI media. In certain embodiments, incubation of the NK cells and antibody is performed in media comprising, consisting of, or consisting essentially of Click’s media. In certain embodiments, incubation of the NK cells and antibody is performed in media comprising, consisting of, or consisting essentially of RPMI media. In certain embodiments, incubation of the NK cells and antibody is performed in media comprising, consisting of, or consisting essentially of SCGM media. In certain embodiments, incubation of the NK cells and antibody is performed in vivo, such as in the blood, lymph, and / or tumor of a subject.

[0260] In certain embodiments, loading of an NK cell can comprise in-vivo loading of the NK cells. In some embodiments, a subject can be administered one or more doses of an antibody prior to administration of an NK cell. In some embodiments, a subject can be administered one or more doses of an antibody after administration of an NK cell. In some embodiments, a subject can be administered one or more doses of an antibody before administration of a loaded NK cell, and a subject can be administered one or more doses of an antibody after administration of a loaded NK cell.

[0261] In some embodiments, loading of an NK cell can comprise providing a subject with at least, exactly, or 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 mg of an antibody, one or more times prior to administration of an NK cell. In some embodiments, loading of an NK cell can comprise providing a subject with at least, exactly, or about 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, 860, 880, 900, 920, 940, 960, 980, or 1000 mg of an antibody, one or more times prior to administration of an NK cell.

[0262] In some embodiments, loading of an NK cell can comprise providing a subject with at least, exactly, or 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 mg of an antibody, one or more times after administration of an NK cell. In some embodiments, loading of an NK cell can comprise providing a subject with at least, exactly, or about 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, 860, 880, 900, 920, 940, 960, 980, or 1000 mg of an antibody, one or more times after administration of an NK cell.

[0263] In some embodiments, loading of an NK cell can comprise providing a subject with at least, exactly, or 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 mg of an antibody, at the same time as administration of an NK cell. In some embodiments, loading of an NK cell can comprise providing a subject with at least, exactly, or about 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, 860, 880, 900, 920, 940, 960, 980, or 1000 mg of an antibody, at the same time as administration of an NK cell.

[0264] The antibodies of the compositions are subjected in an effective amount to an effective amount of NK cells of the disclosure, thereby producing a complex that is “chimeric antigen receptor-like.” In particular, an antigen binding domain of the antibody binds to the NK cells, such as through the antigen that is a cell surface protein. A plurality of antibodies may be subjected to a plurality of NK cells such that there are multiple complexes of cell / antibody. The antibodies may be of any time, including monospecific, bispecific, or multispecific, and in specific cases the antibody engages both the NK cell and a target antigen through an antigen binding domain of the antibody (such as with engagers in the art that are fusion proteins consisting of two single-chain variable fragments (scFvs) of different antibodies). In examples wherein the antibody is monospecific, an antigen binding domain of the antibody binds a target antigen, such as a cancer antigen, and another part of the antibody binds the NK cells, such as an Fc region of the antibody. In cases wherein the antibody is multi specific, one or more antigen binding domains of the antibody may bind the NK cell (such as through an NK cell surface antigen) and one or more antigen binding domains of the antibody binds one or more target antigens. In certain embodiments of cases of use of multispecific antibodies, one or more antigen binding domains of the antibody and / or the Fc region of the antibody may bind the NK cell. In certain embodiments, one or more Fc regions of the antibody may bind the NK cell through a polypeptide encoded by a transgenic construct. The multispecific antibody may be bispecific, trispecific, or tetraspecific, for example. In cases wherein the antibody is trispecific or tetraspecific, the additional antigen binding domains may bind other cells, such as stem cells.

[0265] In particular embodiments, the antibodies may bind any NK cell surface antigen (that may or may not be receptors) on NK cells, such as CD 16 (including CD 16a or CD 16b), CD32, CD56, CD64, a c-type lectin such as NKG2D, NKG2C, a costimulatory molecule such as CS1, DNAM, 2B4, CD2, an NCR, NKp30, NKp44, NKp46, or KIR, and redirect the NK cells to a target, thus increasing the response and specificity against different tumors.

[0266] In some embodiments, the antibodies may bind any suitable antigen (e.g., antigens described herein, such as those that are described as targets of TCRs, etc.). In particularembodiments, an antibody targets CD123. In particular embodiments, an antibody targets EGFR. In particular embodiments, an antibody targets EGFR2. In particular embodiments, an antibody is bi-specific and targets EGFR and c-MET. In particular embodiments, an antibody is Imgatuzumab, Amivantamab, and / or Margetuximab.

[0267] Generation of the complexes may be by any suitable means, such that the conditions are sufficient for the appropriate region of the antibody to bind the appropriate surface region of the NK cell. Any particular medium may be utilized, in certain instances. In specific cases, Plasma-Lyte A and / or human serum albumin are utilized, wherein in other cases they are not. Once the complexes are formed in culture, they may or may not be washed prior to administration to the subject, such as through infusion. In some embodiments, the NK cells and the antibodies are administered separately, and the complexes form in vivo. In certain embodiments, the NK cells and the antibodies are administered separately, and are also administered together, and complexes form in vitro and in vivo. In certain embodiments, a composition comprising NK cells and antibody is washed with PBS to remove unbound antibody. In certain embodiments, a composition comprising NK cells and antibody is washed at least 1, 2, 3, 4, or 5, or greater than 5 times to remove unbound antibody. In certain embodiments, a composition comprising NK cells and antibody is was for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or greater than 10 minutes to remove unbound antibody. In certain embodiments, a composition comprising NK cells and antibody is washed twice to remove unbound antibody. In certain embodiments, a wash is for exactly, or about 5 minutes. In certain embodiments, a wash comprises agitation of the composition, e.g., using a cell shaker.F. Pre-Activation

[0268] In some embodiments, the NK cells are pre-activated prior to administration to a recipient individual. The pre-activation step may or may not occur before any expansion step. In specific embodiments, the NK cells are pre-activated with one or more cytokines, and in specific embodiments, the NK cells are pre-activated with one or more of IL-12, IL-15, IL-2, and IL-18 and including two, three, or more. In cases wherein less than all three of IL-12, IL- 15, IL-2, and IL-18 are utilized, it may be that IL-12 and IL-15 but not IL-18; or IL-12 and IL- 18 but not IL-15; or IL-15 and IL-18 but not IL-12. IL-2 may or may not be substituted for IL- 15.

[0269] In particular embodiments, the pre-activation cytokines may be IL-12, IL-15, and IL-18. One or more additional cytokines may be used for the pre-activation step. The pre- activation may be for a short period of time such as 5-72 hours, such as 10-50 hours,particularly 10-20 hours, such as 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours, and specifically about 16 hours in some cases. The pre-activation culture may comprise IL-18 and / or IL-15 at a concentration of 10-100 ng / mL, such as 40-60 ng / mL, particular 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 ng / mL, specifically about 50 ng / mL. In some cases, the pre-activation culture comprises IL-12 at a concentration of 0.1-150 ng / mL, including at a concentration of 1-20 ng / mL, such as a concentration of 10 ng / mL. In alternative embodiments the NK cells may be stimulated with IL-2, or other cytokines that bind the common gamma-chain (e.g., IL-7, IL-21, and others), and this may be in addition to IL-12, IL-15, and IL-18 or as an alternative to one or more of them. In such cases, the pre-activation culture may comprise IL- 12 at a concentration of 0.1-150 ng / mL, such as 0.5-50 ng / mL, particularly 1-20 ng / mL, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ng / mL, specifically about 10 ng / mL.G. Expansion

[0270] In particular embodiments, NK cells are expanded to increase their quantity prior to administration to an individual in need thereof. The expanded cells may or may not be derived from pre-activated NK cells such that a pre-activation step may occur before an expansion step. The NK cell expansion step may be of any suitable such that the NK cell population is expanded, but in specific cases the expansion step utilizes particular one or more reagents, such as in culture, to enhance their expansion. In certain cases the NK cells may not be expanded. IL-2 or IL- 15 or IL- 18 or any combination of the cytokines may be added to the expansion culture before or during expansion. The NK cells can be expanded ex vivo in flasks or in one of several different bioreactor configurations with continuous perfusion of media / additives, in specific embodiments.

[0271] In specific cases, the NK cells (whether pre-activated or not) may be washed (e.g., with PBS or Plasma Lyte or human serum albumin or culture media or combinations thereof) prior to and / or after expansion, such as 1, 2, 3, 4, or 5 times. In some embodiments, cells are washed specifically 3 times. In particular embodiments, the NK cells are expanded in the presence of artificial antigen presenting cells (aAPCs). In particular embodiments, the NK cells are expanded in the presence of fragments of aAPCs. The aAPCs may be engineered to express CD137 ligand and / or a membrane-bound cytokine. The membrane-bound cytokine may be membrane-bound IL-21 (mIL-21) or membrane-bound IL- 15 (mIL-15). In particular embodiments, the aAPCs are engineered to express CD137 ligand and mIL-2L The aAPCs may be derived from cancer cells, such as leukemia cells. The aAPCs may not express endogenous HLA class I, II, or CD Id molecules. They may express ICAM-1 (CD54) and LFA-3 (CD58) or CD48. In particular, the aAPCs may be K562 cells, such as K562 cells engineered to express CD137 ligand and mIL-21. The engineering may be by any method known in the art, such as retroviral transduction, although any viral or non-viral vector may be utilized. The aAPCs may or may not be irradiated. The expansion may be for a particular duration in time, such as for about 2-30 days, such as 3-20 days, particularly 12-16 days, such as 12, 13, 14, 15, 16, 17, 18, or 19 days, specifically about 14 days. The pre-activated NK cells and aAPCs may be present at a ratio ofabout 3: 1-1 :3, such as 2: 1, 1 : 1, 1 :2, specifically about 1 :2. The expansion culture may further comprise one or more cytokines to promote expansion, such as IL-2. The IL-2 may be present at a concentration of about 10-500 U / mL, such as 100-300 U / mL, particularly about 200 U / mL. The IL-2 may be replenished in the expansion culture, including at a certain frequency, such as every 2-3 days. The aAPCs may be added to the culture at least a second time, such as at about 7 days of expansion. Any cytokine(s) used in the pre-activation and / or expansion steps may be recombinant human cytokines.

[0272] In some embodiments, following expansion, the NK cells may be immediately utilized in any manner, such as complexed with one or more antibodies, or they may be stored, such as by cryopreservation. In certain aspects, the cells may be propagated for days, weeks, or months ex vivo as a bulk population within about 1, 2, 3, 4, or 5 days.

[0273] Activated and / or expanded NK cells can secrete type I cytokines, such as interferon- y, tumor necrosis factor-a and granulocyte-macrophage colony-stimulating factor (GM-CSF), which activate both innate and adaptive immune cells as well as other cytokines and chemokines. The measurement of these cytokines can be used to determine the activation status of NK cells. In addition, other methods known in the art for determination of NK cell activation may be used for characterization of the NK cells of the present disclosure.

[0274] Thus, with respect to particular pre-activation and expansion aspects of the disclosure, in specific embodiments the NK cells pre-activated with any combination of IL-12, IL15, and / or IL-18 followed by expansion with aAPCs, such as K562 cells expressing mIL-21 and CD 137 ligand, provide a highly potent cellular product. Thus, methods are provided using the present NK cells for the treatment of various diseases, such as immunotherapy of patients with cancer. In an exemplary method, the isolated NK cells may be subjected to a brief period, such as about 16 hours, of pre-activation with a combination of cytokines, such as interleukin- 12 (IL-12), IL-15, and / or IL-18, followed by expansion using artificial antigen presenting cells (aAPCs), such as K562 feeder cells expressing membrane-bound IL-21 and CD137 ligand, and / or exogenous IL-2. IL-2 or IL- 15 or IL- 18 or any combination of the cytokines may be added to the expansion culture at least a second time.H. Cryopreservation

[0275] In particular cases, NK cells and / or antibodies of the disclosure are preserved in a cry opreservation medium composition comprising at least one cryoprotectant, a serum (human or animal serum) or a non-serum alternative to serum (not human serum or animal serum), and at least one cytokine and / or at least one growth factor. In some cases, the cryoprotectant is dimethyl sulfoxide (DMSO), glycerin, glycerol, hydroxy ethyl starch, or a combination thereof. The non-serum alternative may be of any kind, including at least platelet lysate and / or a blood product lysate (for example, human serum albumin). In embodiments of the composition wherein one or more (including two or more) cytokines are utilized, the cytokine may be a natural or a recombinant or a synthetic protein. At least one of the cytokines may be an Food and Drug Administration (FDA)-approved cytokine. Examples of cytokines and growth factors include at least IL-1, IL-2, IL-3, IL-4, IL-6, IL-7, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, IL- 18, IL-21, IL-22, interferon, tumor necrosis factor, stem cell factor, FLT3-ligand, APRIL, thrombopoietin, erythropoietin, or a combination thereof. For serum embodiments, the serum may be an animal-derived serum, such as human serum (including human AB serum) or bovine serum. DMSO and other cryoprotectants, when utilized may comprise 4-10%, 4-6%, 4-8%, 5- 10%, 5-8%, 6-10%, 6-8%, 8-10%, and so forth, of the composition. For embodiments wherein serum is employed, the serum may comprise 5-99%, 5-95%, 5-90%, 5-85%, 5-80%, 5-75%, 5- 70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5-20%, 5-15%, 5- 10%, 10-99%, 10-95%, 10-90%, 10-85%, 10-80%, 10-75%, 10-70%, 10-65%, 10-60%, 10-55%, 10-50%, 10-45%, 10-40%, 10-35%, 10-30%, 10-25%, 10-20%, 10-15%, 20-99%, 20-95%, 20-90%. 20-85%, 20-80%, 20-75%, 20-70%, 20-65%, 20-60%, 20-55%, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 20-25%, 30-99%, 30-95%, 30-90%, 30-85%, 30-80%, 30-75%, 30-70%, 30-65%, 30-60%, 30-55%, 30-50%, 30-45%, 30-40%, 30-35%, 40-99%, 40-95%, 40-90%, 40-85%, 40-80%, 40-75%, 40-70%, 40-65%, 40-60%, 40-55%, 40-50%, 40-45%, 50-99%, 50-95%, 50-90%, 50-85%, 50-80%, 50-75%, 50-70%, 50-65%, 50-60%, 50-55%, 60-99%, 60-95%, 60-90%, 60-85%, 60-80%, 60-75%, 60-70%, 60-65%, 70-99%, 70-95%, 70-90%, 70-85%, 70-80%, 70-75%, 80-99%, 80-95%, 80-90%, 80-85%, 90-99%, 90-95%, or 95-99% of the composition. The composition may comprise at least or no more than 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of serum. In specific embodiments, the composition comprises platelet lysate that may be at any concentration in the composition, but in certain embodiments the platelet lysate comprises 5-99%, 5-95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5- 50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-99%, 10-95%, 10-90%, 10-85%, 10-80%, 10-75%, 10-70%, 10-65%, 10-60%, 10-55%, 10-50%, 10-45%, 10-40%, 10-35%, 10-30%, 10-25%, 10-20%, 10-15%, 20-99%, 20-95%, 20-90%. 20-85%, 20-80%, 20-75%, 20-70%, 20-65%, 20-60%, 20-55%, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 20-25%, 30-99%, 30-95%, 30-90%, 30-85%, 30-80%, 30-75%, 30-70%, 30-65%, 30-60%, 30-55%, 30-50%, 30-45%, 30-40%, 30-35%, 40-99%, 40-95%, 40-90%, 40-85%, 40-80%, 40-75%, 40-70%, 40-65%, 40-60%, 40-55%, 40-50%, 40-45%, 50-99%, 50-95%, 50-90%, 50-85%, 50-80%, 50-75%, 50-70%, 50-65%, 50-60%, 50-55%, 60-99%, 60-95%, 60-90%, 60-85%, 60-80%, 60-75%, 60-70%, 60-65%, 70-99%, 70-95%, 70-90%, 70-85%, 70-80%, 70-75%, 80-99%, 80-95%, 80-90%, 80-85%, 90-99%, 90-95%, or 95-99% of the composition. The composition may comprise at least or no more than 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of platelet lysate.

[0276] The composition may have certain concentrations of components, including cytokines and / or growth factors. In specific cases, any cytokine, including IL-2, IL-21, and / or IL- 15, for example, are present in the composition in a particular concentration. The IL-2 may be present at a concentration of 1-5000, 1-1000, 1-500, 1-100, 100-5000, 100-500, 500-5000, 500-1000, or 1000-5000 U / mL, for example. In a specific case, the IL-2 is present at a concentration in the composition of at least or no more than 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 U / mL. In specific embodiments, IL-21 is present in the composition at a concentration of 10-3000, 10-2000, 10-1000, 10-500, 10-100, 100-3000, 100-2000, 100-1000, 500-3000, 500-2000, 500-1000, 1000-3000, 1000-2000, or 2000-3000 ng / mL. The IL-21 may be in a concentration in the composition of at least or nor more than 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 750, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, or 3000 ng / mL. IL-15 may be present in the composition at a concentration of 1-2000, 1-1000, 1-500, 1-100, 100-2000, 100-1000, 100-500, 500-2000, 500-1000, or 1000- 2000 ng / mL. IL-15 may be present in the composition at a concentration of at least or no more than 10, 50, 100, 500, 1000, 1500, or 2000 ng / mL.

[0277] Compositions as encompassed herein that comprise at least one cryoprotectant, a serum or a non-serum alternative to serum, and at least one cytokine and / or at least one growth factor may further comprise a plurality of immune cells and / or stem cells, each of any kind. In specific embodiments, the cells are NK cells, T cells, B cells, NKT cells derived from mature bone marrow or peripheral blood cells, cell lines such as tumor cell lines (e.g., NK92 or other NK lines), hematopoietic stem cells, induced pluripotent stem cells, MSCs (a population of cells alternatively called “mesenchymal stem cells” and “mesenchymal stromal cells” in the literature), or a mixture thereof, which can be derived from bone marrow, peripheral blood,skin, adipose tissue, or a combination thereof. In embodiments wherein NK cells are utilized, the NK cells may or may not be expanded NK cells. Embodiments of the disclosure also encompass pharmaceutical compositions that comprise any composition of the disclosure and a suitable pharmaceutically acceptable carrier.

[0278] In certain embodiments, cells and / or antibodies are treated with one or more deactivating agents (e.g., a kinase inhibitor, e.g., Dasatinib, Nilotinib, Rapamycin, etc.) precryopreservation.

[0279] In some embodiments, technologies described herein comprise deactivating a NK cell, comprising treating an NK cell with an effective amount of one or more deactivating agents under conditions to produce a deactivated NK cell. In some embodiments, a deactivating agent is a kinase inhibitor. In some embodiments, a deactivating agent is a mechanistic target of rapamycin (mTOR) inhibitor. In some embodiments, the mTOR inhibitor is rapamycin, everolimus, and / or temsirolimus. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the deactivating agent is a tyrosine kinase (TK) inhibitor. In some embodiments, the TK inhibitor is Lorlatinib, Brigatinib, Ceritinib, Alectinib, Crizotinib, Bosutinib, Ponatinib, Nilotinib, Dasatinib, Imatinib, Zanubrutinib, Acalabrutinib, Ibrutinib, Capmatinib, Pexidartinib, Dacomitinib, Osimertinib, Erlotinib, Gefitinib, Lapatinib, Afatinib, Pemigatinib, Erdafitinib, Nintedanib, Gilteritinib, Midostaurin, Tucatinib, Neratinib, Baricitinib, Ruxolitinib, Fedratinib, Tofacitinib, Ripretinib, Selumetinib, Binimetinib, Cobimetinib, Trametinib, Upadacitinib, Avapritinib, Selpercatinib, Cabozantinib, Fostamatinib, Larotrectinib, Entrectinib, Axitinib, Regorafenib, Pazopanib, Sorafenib, Lenvatinib, Vandetanib, and / or Sunitinib. In some embodiments, the TK inhibitor is a BCR- Abl inhibitor. In some embodiments, the TK inhibitor is Bosutinib, Ponatinib, Nilotinib, Dasatinib, and / or Imatinib. In some embodiments, the TK inhibitor is Dasatinib and / or Nilotinib. In some embodiments, the TK inhibitor is Dasatinib.

[0280] In some embodiments, treatment with a deactivating agent is at any point during culturing of the NK cell. In some embodiments, the treatment is for about 24 to about 96 hours, about 36 to about 84 hours, or about 48 to about 72 hours. In some embodiments, the treatment is for about 24 hours, about 48 hours, or about 72 hours. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 1 to about 1000 nM. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 5 to about 500 nM. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 20 to about 200 nM. In some embodiments, the NK cell is treated with the deactivating agent at a concentration of about 30 to about 100 nM. In some embodiments,the deactivated NK cell has an increased expression of one or more of C-kit, CCR-5, CD62L and / or CXCR4, and / or decreased expression of one or more of NKG2D, DNAM, OX-40, TRAIL, HLA-DR, CD2, CD25, ICOS, and / or CD95 relative to an activated NK cell.In some embodiments, technologies described herein comprise methods of maintaining the viability of a population of cells over at least 50% percent following cryopreservation of the population, comprising the step of subjecting the population to an effective amount of one or more deactivating agents (e.g., a tyrosine kinase inhibitor) to deactivate the cells prior to cryopreservation, cryopreserving the cells, and thawing the population, wherein upon thawing the viability of the population is over at least 50%. In some cases, upon thawing of the cells the viability of the population of cells is over at least 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% following cryopreservation of the population.III. Heterologous Proteins and Mutations

[0281] In specific embodiments, the NK cells are modified not only to express one or more components of the TCR / CD3 complex and an Fc binding protein, but they are also modified to express one or more other heterologous proteins. The heterologous proteins may facilitate activity of the NK cells in any manner, including at least their activation, persistence, expansion, homing, and / or cytotoxicity.A. Monospecific, Bispecific, or Multi-specific Antibodies

[0282] Aspects of the disclosure relate to use of antibodies or functional fragments thereof in a composition also comprising certain NK cells. The term “antibody” refers to an intact immunoglobulin of any isotype, or a fragment thereof that can compete with the intact antibody for specific binding to the target antigen, such as through the antigen binding domain, and includes chimeric, humanized, fully human, monospecific, and multispecific (including at least bispecific and trispecific, and more) antibodies. As used herein, the terms “antibody” or “immunoglobulin” are used interchangeably and refer to any of several classes of structurally related proteins that function as part of the immune response of an animal, including IgG, IgD, IgE, IgA, IgM, and related proteins, as well as polypeptides comprising antibody CDR domains that retain antigen-binding activity. In specific embodiments, the antibody comprises an scFv. In certain embodiments, the antibodies may be any antibody or antibody-like structure known in the art, including antibody fragments, single-domain antibodies, scFv, bispecific antibodies, bispecific diabodies, trispecific antibodies, scFv-Fc and other antibody constructs and engagers.

[0283] The term “antigen” refers to a molecule or a portion of a molecule capable of being bound by a selective binding agent, such as an antibody. An antigen may possess one or more epitopes that are capable of interacting with different antibodies.

[0284] The term “epitope” includes any region or portion of molecule capable eliciting an immune response by binding to an immunoglobulin or to a T-cell receptor. Epitope determinants may include chemically active surface groups such as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and may have specific three-dimensional structural characteristics and / or specific charge characteristics. Generally, antibodies specific for a particular target antigen will preferentially recognize an epitope on the target antigen within a complex mixture.

[0285] The epitope regions of a given polypeptide can be identified using many different epitope mapping techniques are well known in the art, including: x-ray crystallography, nuclear magnetic resonance spectroscopy, site-directed mutagenesis mapping, protein display arrays, see, e.g., Epitope Mapping Protocols, (Johan Rockb erg and Johan Nilvebrant , Ed., 2018) Humana Press, New York, N.Y. Such techniques are known in the art and described in, e.g., U.S. Pat. No. 4,708,871; Geysen et al. Proc. Natl. Acad. Sci. USA 81:3998-4002 (1984); Geysen et al. Proc. Natl. Acad. Sci. USA 82: 178-182 (1985); Geysen et al. Molec. Immunol. 23:709-715 (1986 See, e.g., Epitope Mapping Protocols, supra. Additionally, antigenic regions of proteins can also be predicted and identified using standard antigenicity and hydropathy plots.

[0286] An intact antibody is generally composed of two full-length heavy chains and two full-length light chains, but in some instances may include fewer chains, such as antibodies naturally occurring in camelids that may comprise only heavy chains. Antibodies as disclosed herein may be derived solely from a single source or may be “chimeric,” that is, different portions of the antibody may be derived from two different antibodies. For example, the variable or CDR regions may be derived from a rat or murine source, while the constant region is derived from a different animal source, such as a human. The antibodies or binding fragments may be produced in hybridomas, by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact antibodies. Unless otherwise indicated, the term “antibody” includes derivatives, variants, fragments, and muteins thereof, examples of which are described below (Sela-Culang et al. Front Immunol. 2013; 4: 302; 2013)

[0287] The term “light chain” includes a full-length light chain and fragments thereof having sufficient variable region sequence to confer binding specificity. A full-length light chain has a molecular weight of around 25,000 Daltons and includes a variable region domain(abbreviated herein as VL), and a constant region domain (abbreviated herein as CL). There are two classifications of light chains, identified as kappa (K) and lambda ( ). The term “VL fragment” means a fragment of the light chain of a monoclonal antibody that includes all or part of the light chain variable region, including CDRs. A VL fragment can further include light chain constant region sequences. The variable region domain of the light chain is at the amino-terminus of the polypeptide.

[0288] The term “heavy chain” includes a full-length heavy chain and fragments thereof having sufficient variable region sequence to confer binding specificity. A full-length heavy chain has a molecular weight of around 50,000 Daltons and includes a variable region domain (abbreviated herein as VH), and three constant region domains (abbreviated herein as CHI, CH2, and CH3). The term “VH fragment” means a fragment of the heavy chain of a monoclonal antibody that includes all or part of the heavy chain variable region, including CDRs. A VH fragment can further include heavy chain constant region sequences. The number of heavy chain constant region domains will depend on the isotype. The VH domain is at the aminoterminus of the polypeptide, and the CH domains are at the carboxy-terminus, with the CH3 being closest to the — COOH end. The isotype of an antibody can be IgM, IgD, IgG, IgA, or IgE and is defined by the heavy chains present of which there are five classifications: mu (p), delta (5), gamma (y), alpha (a), or epsilon (a) chains, respectively. IgG has several subtypes, including, but not limited to, IgGl, IgG2, IgG3, and IgG4. IgM subtypes include IgMl and IgM2. IgA subtypes include IgAl and IgA2.

[0289] Antibodies can be whole immunoglobulins of any isotype or classification, chimeric antibodies, or hybrid antibodies with specificity to two or more antigens. They may also be fragments (e.g., F(ab')2, Fab', Fab, Fv, and the like), including hybrid fragments. An immunoglobulin also includes natural, synthetic, or genetically engineered proteins that act like an antibody by binding to specific antigens to form a complex. The term antibody includes genetically engineered or otherwise modified forms of immunoglobulins, such as the following:

[0290] The term “monomer” means an antibody containing only one Ig unit. Monomers are the basic functional units of antibodies. The term “dimer” means an antibody containing two Ig units attached to one another via constant domains of the antibody heavy chains (the Fc, or fragment crystallizable, region). The complex may be stabilized by a joining (J) chain protein. The term “multimer” means an antibody containing more than two Ig units attached to one another via constant domains of the antibody heavy chains (the Fc region). The complex may be stabilized by a joining (J) chain protein.

[0291] The term “bivalent antibody” means an antibody that comprises two antigenbinding sites. The two binding sites may have the same antigen specificities or they may be bi- specific, meaning the two antigen-binding sites have different antigen specificities.

[0292] Bispecific antibodies are a class of antibodies that have two paratopes with different binding sites for two or more distinct epitopes. In some embodiments, bispecific antibodies can be biparatopic, wherein a bispecific antibody may specifically recognize a different epitope from the same antigen. In some embodiments, bispecific antibodies can be constructed from a pair of different single domain antibodies termed “nanobodies”. Single domain antibodies are sourced and modified from cartilaginous fish and camelids. Nanobodies can be joined together by a linker using techniques typical to a person skilled in the art; such methods for selection and joining of nanobodies are described in PCT Publication No. WO2015044386A1, No. W02010037838A2, and Bever et al., Anal Chem. 86:7875-7882 (2014), each of which are specifically incorporated herein by reference in their entirety.

[0293] Bispecific antibodies can be constructed as: a whole IgG, Fab'2, Fab'PEG, a diabody, or alternatively as scFv. Diabodies and scFvs can be constructed without an Fc region, using only variable domains, potentially reducing the effects of anti -idiotypic reaction. Bispecific antibodies may be produced by a variety of methods including, but not limited to, fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai and Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148: 1547-1553 (1992), each of which are specifically incorporated by reference in their entirety.

[0294] In certain aspects, the antigen-binding domain may be multispecific or heterospecific by multimerizing with VH and VL region pairs that bind a different antigen. For example, the antibody may bind to, or interact with, (a) a cell surface antigen, (b) an Fc receptor on the surface of an effector cell, or (c) at least one other component. Accordingly, aspects may include, but are not limited to, bispecific, trispecific, tetraspecific, and other multispecific antibodies or antigen-binding fragments thereof that are directed to epitopes and to other targets, such as Fc receptors on effector cells.

[0295] In some embodiments, multispecific antibodies can be used and directly linked via a short flexible polypeptide chain, using routine methods known in the art. One such example is diabodies that are bivalent, bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain, and utilize a linker that is too short to allow for pairing between domains on the same chain, thereby forcing the domains to pair with complementary domains of another chain creating two antigen binding sites. The linker functionality is applicable for embodiments of triabodies, tetrabodies, and higher order antibody multimers.(see, e.g., Hollinger et al., Proc Natl. Acad. Sci. USA 90:6444-6448 (1993); Polijak et al., Structure 2: 1121-1123 (1994); Todorovska et al., J. Immunol. Methods 248:47-66 (2001)).

[0296] Bispecific diabodies, as opposed to bispecific whole antibodies, may also be advantageous because they can be readily constructed and expressed in E. coli. Diabodies (and other polypeptides such as antibody fragments) of appropriate binding specificities can be readily selected using phage display (WO94 / 13804) from libraries. If one arm of the diabody is kept constant, for instance, with a specificity directed against a protein, then a library can be made where the other arm is varied and an antibody of appropriate specificity selected. Bispecific whole antibodies may be made by alternative engineering methods as described in Ridgeway et al., (Protein Eng., 9:616-621, 1996) and Krah et al., (N Biotechnol. 39: 167-173, 2017), each of which is hereby incorporated by reference in their entirety.

[0297] Heteroconjugate antibodies are composed of two covalently linked monoclonal antibodies with different specificities. See, e.g., US Patent No. 6,010,902, incorporated herein by reference in its entirety.

[0298] The part of the Fv fragment of an antibody molecule that binds with high specificity to the epitope of the antigen is referred to herein as the “paratope.” The paratope consists of the amino acid residues that make contact with the epitope of an antigen to facilitate antigen recognition. Each of the two Fv fragments of an antibody is composed of the two variable domains, VH and VL, in dimerized configuration. The primary structure of each of the variable domains includes three hypervariable loops separated by, and flanked by, Framework Regions (FR). The hypervariable loops are the regions of highest primary sequences variability among the antibody molecules from any mammal. The term hypervariable loop is sometimes used interchangeably with the term “Complementarity Determining Region (CDR).” The length of the hypervariable loops (or CDRs) varies between antibody molecules. The framework regions of all antibody molecules from a given mammal have high primary sequence similarity / consensus. The consensus of framework regions can be used by one skilled in the art to identify both the framework regions and the hypervariable loops (or CDRs) which are interspersed among the framework regions. The hypervariable loops are given identifying names which distinguish their position within the polypeptide, and on which domain they occur. CDRs in the VL domain are identified as LI, L2, and L3, with LI occurring at the most distal end and L3 occurring closest to the CL domain. The CDRs may also be given the names CDR-1, CDR-2, and CDR-3. The L3 (CDR-3) is generally the region of highest variability among all antibody molecules produced by a given organism. The CDRs are regions of the polypeptide chain arranged linearly in the primary structure, and separated from each other byFramework Regions. The amino terminal (N-terminal) end of the VL chain is named FR1. The region identified as FR2 occurs between LI and L2 hypervariable loops. FR3 occurs between L2 and L3 hypervariable loops, and the FR4 region is closest to the CL domain. This structure and nomenclature is repeated for the VH chain, which includes three CDRs identified as Hl, H2 and H3. The majority of amino acid residues in the variable domains, or Fv fragments (VH and VL), are part of the framework regions (approximately 85%). The three dimensional, or tertiary, structure of an antibody molecule is such that the framework regions are more internal to the molecule and provide the majority of the structure, with the CDRs on the external surface of the molecule.

[0299] Several methods have been developed and can be used by one skilled in the art to identify the exact amino acids that constitute each of these regions. This can be done using any of a number of multiple sequence alignment methods and algorithms, which identify the conserved amino acid residues that make up the framework regions, therefore identifying the CDRs that may vary in length but are located between framework regions. Three commonly used methods have been developed for identification of the CDRs of antibodies: Kabat (as described in T. T. Wu and E. A. Kabat, “AN ANALYSIS OF THE SEQUENCES OF THE VARIABLE REGIONS OF BENCE JONES PROTEINS AND MYELOMA LIGHT CHAINS AND THEIR IMPLICATIONS FOR ANTIBODY COMPLEMENTARITY,” J Exp Med, vol. 132, no. 2, pp. 211-250, Aug. 1970); Chothia (as described in C. Chothia et al., “Conformations of immunoglobulin hypervariable regions,” Nature, vol. 342, no. 6252, pp. 877-883, Dec. 1989); and IMGT (as described in M.-P. Lefranc et al., “IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains,” Developmental & Comparative Immunology, vol. 27, no. 1, pp. 55-77, Jan. 2003). These methods each include unique numbering systems for the identification of the amino acid residues that constitute the variable regions. In most antibody molecules, the amino acid residues that actually contact the epitope of the antigen occur in the CDRs, although in some cases, residues within the framework regions contribute to antigen binding.

[0300] One skilled in the art can use any of several methods to determine the paratope of an antibody. These methods include: 1) Computational predictions of the tertiary structure of the antibody / epitope binding interactions based on the chemical nature of the amino acid sequence of the antibody variable region and composition of the epitope; 2) Hydrogendeuterium exchange and mass spectroscopy; 3) Polypeptide fragmentation and peptide mapping approaches in which one generates multiple overlapping peptide fragments from the full length of the polypeptide and evaluates the binding affinity of these peptides for theepitope; 4) Antibody Phage Display Library analysis in which the antibody Fab fragment encoding genes of the mammal are expressed by bacteriophage in such a way as to be incorporated into the coat of the phage. This population of Fab expressing phage are then allowed to interact with the antigen which has been immobilized or may be expressed in by a different exogenous expression system. Non-binding Fab fragments are washed away, thereby leaving only the specific binding Fab fragments attached to the antigen. The binding Fab fragments can be readily isolated and the genes which encode them determined. This approach can also be used for smaller regions of the Fab fragment including Fv fragments or specific VH and VL domains as appropriate.

[0301] In certain aspects, affinity matured antibodies are enhanced with one or more modifications in one or more CDRs thereof that result in an improvement in the affinity of the antibody for a target antigen as compared to a parent antibody that does not possess those alteration(s). Certain affinity matured antibodies will have nanomolar or picomolar affinities for the target antigen. Affinity matured antibodies are produced by procedures known in the art, e.g., Marks et al., Bio / Technology 10:779 (1992) describes affinity maturation by VH and VL domain shuffling, random mutagenesis of CDR and / or framework residues employed in phage display is described by Rajpal et al., PNAS. 24: 8466-8471 (2005) and Thie et al., Methods Mol Biol. 525:309-22 (2009) in conjugation with computation methods as demonstrated in Tiller et al., Front. Immunol. 8:986 (2017).

[0302] Chimeric immunoglobulins are the products of fused genes derived from different species; “humanized” chimeras generally have the framework region (FR) from human immunoglobulins and one or more CDRs are from a non-human source.

[0303] In certain aspects, portions of the heavy and / or light chain are identical or homologous to corresponding sequences from another particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity. U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81 :6851 (1984). For methods relating to chimeric antibodies, see, e.g., U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81 :6851- 6855 (1985), each of which are specifically incorporated herein by reference in their entirety. CDR grafting is described, for example, in U.S. Pat. Nos. 6,180,370, 5,693,762, 5,693,761, 5,585,089, and 5,530,101, which are all hereby incorporated by reference for all purposes.

[0304] In some embodiments, minimizing the antibody polypeptide sequence from the non-human species optimizes chimeric antibody function and reduces immunogenicity. Specific amino acid residues from non-antigen recognizing regions of the non-human antibody are modified to be homologous to corresponding residues in a human antibody or isotype. One example is the “CDR-grafted” antibody, in which an antibody comprises one or more CDRs from a particular species or belonging to a specific antibody class or subclass, while the remainder of the antibody chain(s) is identical or homologous to a corresponding sequence in antibodies derived from another species or belonging to another antibody class or subclass. For use in humans, the V region composed of CDR1, CDR2, and partial CDR3 for both the light and heavy chain variance region from a non-human immunoglobulin, are grafted with a human antibody framework region, replacing the naturally occurring antigen receptors of the human antibody with the non-human CDRs. In some instances, corresponding non-human residues replace framework region residues of the human immunoglobulin. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody to further refine performance. The humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. See, e.g., Jones et al., Nature 321 :522 (1986); Riechmann et al., Nature 332:323 (1988); Presta, Curr. Op. Struct. Biol. 2:593 (1992); Vaswani and Hamilton, Ann. Allergy, Asthma and Immunol. 1 : 105 (1998); Harris, Biochem. Soc. Transactions 23; 1035 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428 (1994); Verhoeyen et al., Science 239: 1534-36 (1988).

[0305] Intrabodies are intracellularly localized immunoglobulins that bind to intracellular antigens as opposed to secreted antibodies, which bind antigens in the extracellular space.

[0306] Polyclonal antibody preparations typically include different antibodies against different determinants (epitopes). In order to produce polyclonal antibodies, a host, such as a rabbit or goat, is immunized with the antigen or antigen fragment, generally with an adjuvant and, if necessary, coupled to a carrier. Antibodies to the antigen are subsequently collected from the sera of the host. The polyclonal antibody can be affinity purified against the antigen rendering it monospecific.

[0307] Monoclonal antibodies or “mAb” refer to an antibody obtained from a population of homogeneous antibodies from an exclusive parental cell, e.g., the population is identical except for naturally occurring mutations that may be present in minor amounts. Each monoclonal antibody is directed against a single antigenic determinant.

[0308] Functional antibody fragments and antigen-binding fragments may be utilized. Certain aspects relate to antibody fragments, such as antibody fragments that bind to and / orneutralize inflammatory mediators. The term functional antibody fragment includes antigenbinding fragments of an antibody that retain the ability to specifically bind to an antigen. These fragments are constituted of various arrangements of the variable region heavy chain (VH) and / or light chain (VL); and in some embodiments, include constant region heavy chain 1 (CHI) and light chain (CL). In some embodiments, they lack the Fc region constituted of heavy chain 2 (CH2) and 3 (CH3) domains. Embodiments of antigen binding fragments and the modifications thereof may include: (i) the Fab fragment type constituted with the VL, VH, CL, and CHI domains; (ii) the Fd fragment type constituted with the VH and CHI domains; (iii) the Fv fragment type constituted with the VH and VL domains; (iv) the single domain fragment type, dAb, (Ward, 1989; McCafferty et al., 1990; Holt et al., 2003) constituted with a single VH or VL domain; (v) isolated complementarity determining region (CDR) regions. Such terms are described, for example, in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, NY (1989); Molec. Biology and Biotechnology: A Comprehensive Desk Reference (Myers, R. A. (ed.), New York: VCH Publisher, Inc.); Huston et al., Cell Biophysics, 22: 189-224 (1993); Pluckthun and Skerra, Meth. Enzymol., 178:497-515 (1989) and in Day, E. D., Advanced Immunochemistry, 2d ed., Wiley-Liss, Inc. New York, N.Y. (1990); Antibodies, 4:259-277 (2015). The citations in this paragraph are all incorporated by reference.

[0309] Antigen-binding fragments also include fragments of an antibody that retain exactly, at least, or at most 1, 2, or 3 complementarity determining regions (CDRs) from a light chain variable region. Fusions of CDR-containing sequences to an Fc region (or a CH2 or CH3 region thereof) are included within the scope of this definition including, for example, scFv fused, directly or indirectly, to an Fc region are included herein.

[0310] The term Fab fragment means a monovalent antigen-binding fragment of an antibody containing the VL, VH, CL and CHI domains. The term Fab' fragment means a monovalent antigen-binding fragment of a monoclonal antibody that is larger than a Fab fragment. For example, a Fab' fragment includes the VL, VH, CL and CHI domains and all or part of the hinge region. The term F(ab')2 fragment means a bivalent antigen-binding fragment of a monoclonal antibody comprising two Fab' fragments linked by a disulfide bridge at the hinge region. An F(ab')2 fragment includes, for example, all or part of the two VH and VL domains, and can further include all or part of the two CL and CHI domains.

[0311] The term Fd fragment means a fragment of the heavy chain of a monoclonal antibody, which includes all or part of the VH, including the CDRs. An Fd fragment can further include CHI region sequences.

[0312] The term Fv fragment means a monovalent antigen-binding fragment of a monoclonal antibody, including all or part of the VL and VH, and absent of the CL and CHI domains. The VL and VH include, for example, the CDRs. Single-chain antibodies (sFv or scFv) are Fv molecules in which the VL and VH regions have been connected by a flexible linker to form a single polypeptide chain, which forms an antigen-binding fragment. Single chain antibodies are discussed in detail in International Patent Application Publication No. WO 88 / 01649 and U.S. Pat. Nos. 4,946,778 and 5,260,203, the disclosures of which are herein incorporated by reference. The term (scFv)2 means bivalent or bispecific sFv polypeptide chains that include oligomerization domains at their C-termini, separated from the sFv by a hinge region (Pack et al. 1992). The oligomerization domain comprises self-associating a- helices, e.g., leucine zippers, which can be further stabilized by additional disulfide bonds. (scFv)2 fragments are also known as “miniantibodies” or “minibodies.”

[0313] A single domain antibody is an antigen-binding fragment containing only a VH or the VL domain. In some instances, two or more VH regions are covalently joined with a peptide linker to create a bivalent domain antibody. The two VH regions of a bivalent domain antibody may target the same or different antigens.

[0314] In some cases, fragment crystallizable regions, Fc, are utilized. An Fc region contains two heavy chain fragments comprising the CH2 and CH3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domains. The term “Fc polypeptide” as used herein includes native and mutein forms of polypeptides derived from the Fc region of an antibody. Truncated forms of such polypeptides containing the hinge region that promotes dimerization are included. In some embodiments, antibodies that incorporate cytokines (including those referred to herein) may be utilized (such as with TRIKEs).

[0315] In some embodiments, an antibody is an antibody-drug conjugate. In some embodiments, an antibody may specifically be one or more of the following, although an antibody is not limited to these: [fam] -trastuzumab deruxtecan, Abciximab, Adalimumab, Ado- trastuzumab emtansine, Aducanumab, Alemtuzumab, Alirocumab, Amivantamab, Anifrolumab, Ansuvimab, Atezolizumab, Atoltivimab with Maftivimab and Odesivimab-ebgn (aka Inmazeb), Avelumab, Basiliximab, Belantamab mafodotin, Belimumab, Benralizumab, Bevacizumab, Bezlotoxumab, Bimekizumab, Blinatumomab, Brentuximab vedotin, Brodalumab, Brolucizumab, Burosumab, Canakinumab, Caplacizumab, Casirivimab + imdevimab, Catumaxomab, Cemiplimab, Certolizumab pegol, Cetuximab, Cevostamab, Crizanlizumab, Daclizumab, Daratumumab, Denosumab, Dinutuximab, Donanemab,Dostarlimab, Dupilumab, Durvalumab, Eculizumab, Edrecolomab, Efalizumab, Elotuzumab, Emapalumab, Emicizumab, Enfortumab vedotin, Eptinezumab, Erenumab, Evinacumab, Evolocumab, Faricimab, Fremanezumab, Galcanezumab, Gemtuzumab, Gemtuzumab- Ozogamicin, Golimumab, Guselkumab, Ibalizumab, Ibritumomab tiuxetan, Idarucizumab, Imgatuzumab, Inebilizumab, Infliximab, Inolimomab, Inotuzumab, Inotuzumab-Ozogamicin, IPH61, Ipilimumab, Isatuximab, Ixekizumab, Lanadelumab, Lecanemab, Loncastuximab tesirine, Margetuximab, Mepolizumab, Mirvetuximab soravtansine, Mogamulizumab, Mosunetuzumab, Moxetumomab pasudotox, Murom onab-CD3, Narsoplimab, Natalizumab, Naxitamab, ...

Claims

WHAT IS CLAIMED IS:

1. A polynucleotide comprising a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to transcriptional reading frames comprising SEQ ID NO: 171-175.

2. The polynucleotide of claim 1, wherein the polynucleotide is comprised in a vector comprising at least about 80%, 85%, 90%, 95%, 98%, or 100% sequence identity to SEQ ID NOs: 177-181.

3. A polynucleotide comprising a sequence encoding a T cell receptor (TCR) alpha and TCR beta polypeptide, and / or TCR gamma and TCR delta polypeptide, and a polypeptide comprising a CD16 derived Fc binding domain.

4. The polynucleotide of claim 3, wherein the TCR polypeptides are invariant TCR (iTCR) polypeptides.

5. The polynucleotide of claim 4, wherein the iTCRa and iTCRP polypeptides and / or polynucleotides encoding the same comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 51-149.

6. The polynucleotide of claim 4, wherein the iTCRP polypeptide comprises a polynucleotide encoding a VP-DJ region that is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 75-149.

7. The polynucleotide of claim 4, wherein the iTCRP polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NO: NOs: 59-74.

8. The polynucleotide of claim 4, wherein the encoded iTCRP polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 60.

9. The polynucleotide of claim 8, wherein the polynucleotide encoding the iTCRP polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 59.

10. The polynucleotide of claim 4, wherein the iTCRa polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NOs: 51-52.

11. The polynucleotide of claim 3, wherein the polypeptide comprising a CD16 derived Fc binding domain comprises a human CD16A Fc binding domain.

12. The polynucleotide of claim 3, further comprising the Fc binding domain being fused in N to C terminus order to an optional hinge domain, a transmembrane domain (TMD), and one or more optional intracellular signaling domains (ICD).

13. The polynucleotide of claim 12, wherein the TMD is derived from CD16, or CD3(^.

14. The polynucleotide of claim 12, wherein the TMD comprises or consists of a sequence with at least about 90% identity SEQ ID NOs: 163 or 167.

15. The polynucleotide of claim 12, wherein the hinge domain is derived from CD32.

16. The polynucleotide of claim 12, comprising a hinge domain that comprises or consists of a sequence with at least about 90% identity to SEQ ID NO: 161.

17. The polynucleotide of claim 12, comprising an ICD derived from CD16 and / or CD3(^.

18. The polynucleotide of claim 3, wherein the polypeptide comprising a CD16 derived Fc binding domain does not comprise a mutation that renders the CD16 derived Fc binding domain resistant to cleavage.

19. The polynucleotide of claim 3, comprising a coding sequence that is least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 150-154.

20. The polynucleotide of claim 3, wherein the polynucleotide further encodes one or more cytokine sequences.

21. The polynucleotide of claim 20, wherein the cytokine comprises IL- 15 and / or IL-21.

22. An engineered NK cell comprising the polynucleotide of any one of claims 1-21.

23. A method of treating a disease in an individual, the method comprising administration of the engineered NK cells of claim 22 to an individual in need thereof.

24. An engineered immune cell comprising, one or more transgenic polynucleotides encoding: a) a CD3 protein complex comprising part or all of a single chain or any combination of CD35, CD3s, CD3y, or CD3^, b) optionally at least one cytokine, c) at least one TCRa and TCRP chain and / or a TCRy and TCR5 chain, and d) a polypeptide comprising a CD 16 Fc binding domain.

25. The engineered immune cell of claim 24, wherein the one or more transgenic polynucleotides comprise multi ci str onic transcriptional open reading frames.

26. The engineered immune cell of claim 24, wherein the cells are modified to express part or all of CD36, two of CD3s, CD3y, and / or CD3<^.

27. The engineered immune cell of claim 24, wherein any one or more of CD36, CD3s, CD3y, and / or CD3<^ are linked to one or more heterologous intracellular signaling domains.

28. The engineered immune cell of claim 27, wherein the heterologous intracellular signaling domain is selected from the group consisting of CD16, NKG2D, DAP10, DAP12, 2B4, 4-1BB, CD2, CD28, and a combination thereof.

29. The engineered immune cell of claim 27, wherein the heterologous intracellular signaling domain comprises a DAP10 intracellular signaling domain.

30. The engineered immune cell of claim 29, wherein the heterologous intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 42.

31. The engineered immune cell of claim 27, wherein the heterologous intracellular signaling domain comprises a CD28 intracellular signaling domain.

32. The engineered immune cell of claim 31, wherein the heterologous intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 43.

33. The engineered immune cell of claim 27, wherein the heterologous intracellular signaling domain comprises a DAP 10 and CD28 intracellular signaling domain.

34. The engineered immune cell of claim 33, wherein the heterologous intracellular signaling domain comprises an amino acid sequence at least about 85% identical to SEQ ID NO: 44.

35. The engineered immune cell of claim 24, wherein coding sequences for a CD3 protein complex and at least one cytokine are comprised in a first multi ci str onic construct, and wherein coding sequences for at least one TCRa and TCRP chain and / or a TCRy and TCR5 chain, and the polypeptide comprising a CD16 Fc binding domain are encoded by a second multi ci stronic construct.

36. The engineered immune cell of claim 24, comprising a coding sequence for a cytokine, wherein the cytokine comprises IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, IL-23, and / or GMCSF.

37. The engineered immune cell of claim 36, where the cell is modified to express a polynucleotide sequence at least 85% identical to UT-NK15-DAP10 (SEQ ID NO: 45), UT- NK15-28 (SEQ ID NO: 47), or UTNK15-28-DAP10 (SEQ ID NO: 49).

38. The engineered immune cell of claim 37, wherein the cytokine comprises IL-15 and / or IL- 21.

39. The engineered immune cell of claim 38, wherein the cytokine comprises IL-15 and comprises a polypeptide sequence and / or polynucleotide sequence encoding the same that is at least 85% identical to any one or more of SEQ ID NOs: 182-183.

40. The engineered immune cell of claim 38, wherein the cytokine comprises IL-21 and comprises a polypeptide sequence and / or polynucleotide sequence encoding the same that is at least 85% identical to any one or more of SEQ ID NOs: 184-187.

41. The engineered immune cell of claim 24, wherein the TCR polypeptides are invariant TCR (iTCR) polypeptides, and wherein the iTCRa and iTCRP polypeptides and / or polynucleotides encoding the same comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 51-149.

42. The engineered immune cell of any one of claims 24 to 41, wherein the TCR polypeptides are invariant TCR (iTCR) polypeptides.

43. The engineered immune cell of claim 42, wherein the iTCRa and iTCRP polypeptides and / or polynucleotides encoding the same comprise a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 51-149.

44. The engineered immune cell of claim 42, wherein the iTCRP polypeptide comprises a polynucleotide encoding a VP-DJ region that is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 75-149.

45. The engineered immune cell of claim 42, wherein the iTCRP polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NO: NOs: 59-74.

46. The engineered immune cell of claim 42, wherein the encoded iTCRP polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 60.

47. The engineered immune cell of claim 46, wherein the polynucleotide encoding the iTCRP polypeptide is at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 59.

48. The engineered immune cell of claim 42, wherein the iTCRa polypeptide and / or polynucleotide encoding the same comprises a sequence at least about 80%, 85%, 90%, 95%, 98%, or 100% identical to SEQ ID NO: 51-52.

49. The engineered immune cell of claim 24, wherein the CD16 Fc binding domain comprising polypeptide comprises a human CD 16 derived Fc binding domain.

50. The engineered immune cell of claim 24, wherein the polypeptide comprising a CD 16 derived Fc binding domain comprises a human CD16A Fc binding domain.

51. The engineered immune cell of claim 24, wherein the Fc binding domain is fused in N to C terminus order to an optional hinge domain, a transmembrane domain (TMD), and one or more optional intracellular signaling domains (ICD).

52. The engineered immune cell of claim 51, wherein the TMD is derived from CD16, or CD3(^.

53. The engineered immune cell of claim 51, wherein the TMD comprises or consists of a sequence with at least about 90% identity to SEQ ID NOs: 163 or 167.

54. The engineered immune cell of claim 51, wherein the hinge domain is derived from CD32.

55. The engineered immune cell of claim 51, wherein the hinge domain comprises or consists of a sequence with at least about 90% identity to SEQ ID NO: 161.

56. The engineered immune cell of claim 51, comprising an ICD derived from CD 16 and / or CD357. The engineered immune cell of claim 24, comprising a coding sequence that is least about 80%, 85%, 90%, 95%, 98%, or 100% identical to any one or more of SEQ ID NOs: 171-175.

58. The engineered immune cell of any one of claims 24 to 41, wherein the cell is a Natural Killer (NK) cell.

59. The engineered NK cell of claim 58, wherein the NK cells are derived from cord blood (CB), peripheral blood (PB), bone marrow, stem cells, or a combination thereof.

60. The engineered NK cell of claim 58, wherein the NK cells are primary NK cells, and are not derived from stem cells and / or induced pluripotent stem cells (iPSCs).

61. The engineered NK cell of claim 58, wherein the NK cells are loaded with (complexed with) one or more antibodies.

62. The engineered NK cell of claim 61, wherein the one or more antibodies are one or more monospecific, bispecific, or multi-specific antibodies.

63. The engineered NK cell of claim 61, wherein at least one or more antibodies comprises a glycoengineered Fc domain that has a high affinity to wild type CD16.

64. The engineered NK cell of claim 61, wherein the at least one or more antibodies comprise a non-glycoengineered Fc domain that has a low affinity to wild type CD 16 Fc binding domains.

65. The engineered NK cell of claim 64, wherein the non-glycoengineered Fc domain is loaded on (complexed to) the transgenic polypeptide comprising a CD 16 Fc binding domain.

66. The engineered NK cell of claim 61, wherein the one or more antibodies comprise an IgGl and / or IgG4 Fc domain.

67. The engineered NK cell of claim 61 , wherein the one or more antibodies target antigens CD3, CD16, CD28, EGFR, c-MET, CD30, PSMA, MUC17, CD33, FLT3, STEAP1, BCMA, CLDN18.2, CD123, CD19, CD20, EpCAM, CEA, GPC3, CD38, CD33, CD22, HER2, GPA33, GD2, MUC16, GPRC5D, DLL-3, CLEC12A, FcRH5, and / or SSTR.

68. The engineered NK cell of claim 61, wherein the one or more antibodies target CD3, CD16, CD28, CD19, CD20, CD30, HER2, GPRC5D, EGFR, c-MET, and / or BCMA.

69. The engineered NK cell of claim 61, wherein the one or more antibodies comprise [fam]-trastuzumab deruxtecan, Abciximab, Adalimumab, Ado-trastuzumab emtansine, Aducanumab, Alemtuzumab, Alirocumab, Amivantamab, Anifrolumab, Ansuvimab, Atezolizumab, Atoltivimab with Maftivimab and Odesivimab-ebgn (aka Inmazeb), Avelumab, Basiliximab, Belantamab mafodotin, Belimumab, Benralizumab, Bevacizumab, Bezlotoxumab, Bimekizumab, Blinatumomab, Brentuximab vedotin, Brodalumab,Brolucizumab, Burosumab, Canakinumab, Caplacizumab, Casirivimab + imdevimab, Catumaxomab, Cemiplimab, Certolizumab pegol, Cetuximab, Cevostamab, Crizanlizumab, Daclizumab, Daratumumab, Denosumab, Dinutuximab, Donanemab, Dostarlimab, Dupilumab, Durvalumab, Eculizumab, Edrecolomab, Efalizumab, Elotuzumab, Emapalumab, Emicizumab, Enfortumab vedotin, Eptinezumab, Erenumab, Evinacumab, Evolocumab, Faricimab, Fremanezumab, Galcanezumab, Gemtuzumab, Gemtuzumab-Ozogamicin, Golimumab, Guselkumab, Ibalizumab, Ibritumomab tiuxetan, Idarucizumab, Imgatuzumab, Inebilizumab, Infliximab, Inolimomab, Inotuzumab, Inotuzumab-Ozogamicin, IPH61, Ipilimumab, Isatuximab, Ixekizumab, Lanadelumab, Lecanemab, Loncastuximab tesirine, Margetuximab, Mepolizumab, Mirvetuximab soravtansine, Mogamulizumab, Mosunetuzumab, Moxetumomab pasudotox, Murom onab-CD3, Narsoplimab, Natalizumab, Naxitamab, Nebacumab, Necitumumab, Nirsevimab, Nivolumab, Obiltoxaximab, Obinutuzumab, Ocrelizumab, Ofatumumab, Olaratumab, Omalizumab, Omburtamab, Oportuzumab monatox, Palivizumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin, Ramucirumab, Ranibizumab, Ravulizumab, Raxibacumab, Regdanvimab, Relatlimab, Reslizumab, Retifanlimab, Risankizumab, Rituximab, Romosozumab, Sacituzumab govitecan, Sarilumab, Satralizumab, Secukinumab, Siltuximab, Sintilimab, Sotrovimab, Spesolimab, Sutimlimab, Tafasitamab, Tebentafusp, Teclistamab, Teplizumab, Teprotumumab, Tezepelumab, Tildrakizumab, Tislelizumab, Tisotumab vedotin, Tixagevimab, cilgavimab, Tocilizumab, Toripalimab, Tositumomab-1131, Tralokinumab, Trastuzumab, Tremelimumab, Ublituximab, Ustekinumab, Vedolizumab, AMG 160 / Acapatamab, AMG 199 / TNB 585, AMG 330, AMG 427 / EMIRODATAMAB, AMG 509, AMG 701, AMG 910, APVO414 / ES414 / MOR209, APVO436, Catumaxomab / Removab, CC- 1, CC-93269 / EM801, Cibisatamab / RG7802 / RO6958688, CLN-049, Elranatamab / PF- 06863135, EMB-06, Epcoritamab / GEN3013, ERY974, Flotetuzumab / MGD006,Glofitamab / RG6026 / RO7082859, ISB 1342 / GBR 1342, JNJ-63709178, JNJ-63898081, JNJ- 67571244, JNJ-75348780, Linvoseltamab / REGN 5458, M701, M802, MGD007, Mosunetuzumab / RG7828, Nivatrotamab / Hu3F8-BsAb, Odronextamab / REGN1979, REGN4018, REGN5459, REGN7075, REGN5678, Talquetamab / JNJ-64407564,Tarlatamab / AMG 757, Tepoditamab / MCLA-117, TNB-383B, TNB-486, TNB-585, XmAbl3676 / Plamotamab, XmAbl4045 / Vibecotamab, XmAbl8087 / Tidutamab, and / or AFM13.

70. The engineered NK cell of claim 61, wherein the one or more antibodies comprise Elranatamab, Imgatuzumab, Margetuximab, Amivantamab, Blinatumomab, Obinutuzumab, IPH61 (also known as IPH6101 or SAR443579), Teclistamab, Cetuximab, Talquetamab, Pertuzumab, Trastuzumab, Tafasitamab, Brentuximab, and / or Rituximab.

71. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Elranatamab.

72. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Imgatuzumab.

73. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Margetuximab.

74. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Amivantamab.

75. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Blinatumomab.

76. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Obinutuzumab.

77. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of IPH61.

78. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Teclistamab.

79. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Cetuximab.

80. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Rituximab.

81. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Talquetamab.

82. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Pertuzumab.

83. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Trastuzumab.

84. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Tafasitamab.

85. The engineered NK cell of claim 70, wherein the one or more antibodies comprises or consists of Brentuximab.

86. The engineered NK cell of claim 61, wherein the NK cell expresses the one or more antibodies.

87. The engineered NK cell of claim 58, wherein the NK cell is further modified to express one or more additional heterologous proteins selected from the group consisting of an antigen receptor, a cytokine, a homing receptor, a chemokine receptor, and a combination thereof.

88. The engineered NK cell of claim 58, wherein the NK cells are pre-activated with one or more cytokines.

89. The engineered NK cell of claim 88, wherein the cytokines are IL-2, IL-7, IL-12, IL- 15, IL- 18, IL-21, or a combination thereof.

90. The engineered NK cell claim 58, wherein the NK cell further comprises one or more engineered mutations in an endogenous gene.

91. The engineered NK cell of claim 90, wherein the endogenous gene is GR, TGFBR2, CISH, and / or CD38.

92. A composition comprising the engineered NK cell of any one of claims 58 to 91.

93. The composition claim 92, further comprising a pharmaceutically acceptable excipient.

94. The composition claim 92 or 93, wherein the composition is comprised in a delivery device.

95. A method of treating a disease in an individual, the method comprising the step of administering to the individual a therapeutically effective amount of any one of the cells of claim 24 to 41.

96. The method of claim 95, wherein the disease is an autoimmune disease, infection, and / or cancer.

97. The method of claim 96, wherein the disease is an autoimmune disease.

98. The method of claim 97, wherein the autoimmune disease comprises a B cell related autoimmunity, systemic lupus erythematosus (SLE), Systemic scleroderma (SSc), multiple sclerosis (MS), Grave’s disease, rheumatoid arthritis (RA), myositis, dermatomyositis, myasthenia gravis, Sjogren’s syndrome, diffuse scleroderma, inflammatory myopathy, inflammatory myopathy, ANCA-associated systemic vasculitis, antiphospholipid syndrome, immune nephritis, ITP, refractory POEMS syndrome, amyloidosis, autoimmune hemolytic anemia, and / or vasculitis.

99. The method of claim 95, wherein the disease is cancer.

100. The method of claim 99, wherein the cancer expresses CD19, CD20, CD30, HER2, GPRC5D, EGFR, c-MET, and / or BCMA.

101. The method of claim 99, wherein the cancer is pancreatic cancer, colorectal cancer, ovarian cancer, kidney cancer, glioblastoma, breast cancer, renal cancer, myeloma, and / or leukemia.

102. The method of claim 98 or 101, further comprising administering to the individual at the same time or at different time, one or more monospecific, bispecific, and / or multispecific antibodies.

103. The method of claim 102, wherein the one or more antibodies comprise Elranatamab, Imgatuzumab, Margetuximab, Amivantamab, Blinatumomab, Obinutuzumab, IPH61 (also known as IPH6101 or SAR443579), Teclistamab, Cetuximab, Talquetamab, Pertuzumab, Trastuzumab, Tafasitamab, Brentuximab, and / or Rituximab.

104. The method of claim 102, wherein the one or more antibodies are administered at the same time, and / or the one or more antibodies and the engineered NK cells are complexed prior to administration to the individual.

105. The method of claim 102, wherein the one or more antibodies are administered more than once, including at least once at a time point after administration of the engineered NK cells.

106. The method of claim 102, wherein the one or more antibodies are administered more than once, including at least once at a time point before administration of the engineered NK cells.

107. The method of claim 102, wherein the one or more antibodies are administered more than once, including at least once at a time point before administration of the engineered NK cells and at least once at a time point after administration of the engineered NK cells.

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