Pantigen specific cytokine receptor stimulation of immune cell function

Abstract

In aspects, a chimeric cytokine receptor (CCR) is provided that comprises (i) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor domains, an interleukin receptor transmembrane domain, and an interleukin receptor intracellular domain; and (ii) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor common gamma chain (γc) domains, a γc transmembrane domain, and a γc intracellular domain.

Description

PANTIGEN SPECIFIC CYTOKINE RECEPTOR STIMULATION OF IMMUNE CELL FUNCTION

[0001] The present application claims the benefit of U.S provisional application no. 63 / 562,184 filed March 6, 2024, which is incorporated by reference herein in its entirety.FIELD

[0002] The present disclosure relates in general to immunotherapeutic compositions methods of cancer treatment. The disclosure relates in particular to antigen-specific cytokine signaling in immune cells utilizing engineered chimeric cytokine receptor molecules.BACKGROUND

[0003] The incidence of leukemia is rising annually,1with nearly half a million new cases of this poor prognosis malignancy diagnosed globally in 2020.2Acute myeloid leukemia (AML), a leukemia originating from myeloid progenitor cells, is the most common form of acute leukemia in adults.3AML has an aggressive disease course with a 5 -year survival rate of only approximately 30%.4AML therapeutic regimens are associated with systemic toxicities and patients with poor fitness are unable to receive full intensity treatment. Even with rigorous therapy and hematopoietic stem cell transplant, patients relapse. Alternate therapies with less harmful side effects that can be applied broadly are sorely needed.

[0004] Immunotherapies are alternatives to chemotherapies and their extensive side effects. Dramatic anti -cancer efficacy of engineered chimeric antigen receptor (CAR) T cells have led to the FDA-approval of multiple novel products.5However, no such T-cell therapy for AML has been approved, or yet shown efficacy in clinical trial. This is due to T-cell dysfunction at AML diagnosis and immune dysfunction as therapy progresses.6These together contribute to lack of healthy starting material for product manufacture. Allogeneic donor T-cell therapies have been investigated but their use has the associated risk of severe graft-versus-host disease (GVHD).7

[0005] Unlike T cells, allogenic natural killer (NK) cells do not cause GVHD.8This gives NK-cell therapies the potential to be an “off-the-shelf’ cell therapy. Various labs, including ours, have shown NK-cell antigen-specific cytotoxicity can be significantly enhanced using CARs.However, various clinical trials have shown that NK cells are not persistent in vivo following adoptive transfer.9'11Persistence is key for an infused cell product to maintain durable efficacy against leukemic blasts12and to therefore ensure long-lasting remission and cure.

[0006] Interleukin- 15 (IL- 15) is essential forNK-cell development and increases bothNK- cell proliferation and survival.13, 14We have shown that engineering CAR-NK cells with constitutive secreted IL-15 can enhance activation and persistence.15Despite potent anti-AML activity in vitro and in vivo, CAR-NK cells expressing IL- 15 caused lethal toxicity in xenograft mouse models. Clinically, human patients who have received soluble injections of IL-15 or IL-15 complexed with its a receptor display severe inflammatory toxicity.16SUMMARY

[0007] Embodiments are directed to chimeric cytokine receptor molecules and their uses as immunotherapeutics.

[0008] Accordingly, in certain aspects, a chimeric cytokine receptor (CCR) comprises (i) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor domains, an interleukin receptor transmembrane domain, and an interleukin receptor intracellular domain; and (ii) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor common gamma chain (yc) domains, a yc transmembrane domain, and a yc intracellular domain. In certain embodiments, the one or more extracellular interleukin receptor domains comprise one or more interleukin 15 receptor (IL- 15R) extracellular domains or variants thereof. In certain embodiments, the one or more transmembrane interleukin receptor domains comprise one or more interleukin 15 receptor (IL-15R) transmembrane domains or variants thereof. In certain embodiments, the one or more intracellular interleukin receptor domains comprise one or more interleukin 15 receptor (IL- 15R) intracellular domains or variants thereof. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 50% shorter than a wild type IL-15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 75% shorter than a wild type IL-15R extracellular domain. In certain embodiments, the one or moreextracellular domains of the IL-15R comprise an amino acid sequence length that is at least 90% shorter than a wild type IL-15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 95% shorter than a wild type IL-15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 99% shorter than a wild type IL-15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is as long as a wild type IL-15R extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 50% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 75% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 90% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 95% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 99% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is as long as a wild type yc extracellular domain. In certain embodiments, the cytokine receptor comprises an amino acid sequence identity of at least 75% to SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A. In certain embodiments, the cytokine receptor comprises an amino acid sequence identity of at least 80% to SEQ ID NOs: : 2, 4, 4A, 5, 5A, 6 or 6A. In certain embodiments, the cytokine receptor comprises an amino acid sequence identity of at least 90% to SEQ ID NOs: 2, 4, 5 or 6. In certain embodiments, the cytokine receptor comprises an amino acid sequence identity of at least 95% to SEQ ID NOs: : 2, 4, 4A, 5, 5A, 6 or 6A. In certain embodiments, the cytokine receptor comprises SEQ ID NOs: : 2, 4, 4A, 5, 5A, 6 or 6A. In certain embodiments, the cytokine receptor comprises a nucleic acid sequence identity of at least 75% to SEQ ID NOs: 1 or 3. In certain embodiments, the cytokine receptor comprises a nucleic acid sequence identity of at least 80% to SEQ ID NOs: 1 or 3. In certain embodiments, the cytokinereceptor comprises a nucleic acid sequence identity of at least 90% to SEQ ID NOs: 1 or 3. In certain embodiments, the cytokine receptor comprises a nucleic acid sequence identity of at least 95% to SEQ ID NOs: 1 or 3. In certain embodiments, the cytokine receptor comprises SEQ ID NOs: 1 or 3. In certain embodiments, the antigen specific binding domain specifically binds to tumor antigens or virus antigens. In certain embodiments, the antigen specific binding domain specifically binds cluster of differentiation antigen 123 (CD123). In certain embodiments, the antigen specific binding domain comprises an antibody, an antibody binding fragment, an aptamer, or a peptide. In certain embodiments, the antigen specific binding domain comprises a single chain variable fragment (scFv).

[0009] In another aspect, a vector comprises a cytokine receptor nucleic acid sequence having a nucleic acid sequence identity of at least 75% to SEQ ID NOs: 1 or 3. In certain embodiments, the vector comprises a cytokine receptor nucleic acid sequence having a nucleic acid sequence identity of at least 95% to SEQ ID NOs: 1 or 3. In certain embodiments, the vector comprises a cytokine receptor nucleic acid sequence comprising SEQ ID NOs: 1 or 3. In some embodiments, a first nucleic acid sequence (e.g. encoding an IL-15R or variants thereof), and a second nucleic acid sequence (e.g. encoding a yc chain, or variants thereof) are disposed on separate nucleic acid molecules, e.g., separate vectors, e.g., separate viral vectors, e.g., separate retrovirus vectors. In some embodiments, the first nucleic acid sequence is disposed on a first nucleic acid molecule, e.g., a first vector, e.g., a first viral vector, e.g., a first retrovirus vector. In some embodiments, the second nucleic acid sequence is disposed on a second nucleic acid molecule, e.g., a second vector, e.g., a second viral vector, e.g., a second retrovirus vector.

[0010] In some embodiments, a first nucleic acid sequence (e.g. encoding an IL-15R or variants thereof), and a second nucleic acid sequence (e g. encoding a yc chain, or variants thereof) are disposed on the same nucleic acid molecules, e.g., a polycistronic or bicistronic vector. Such polycistronic or bicistronic vectors can be preferred and may provide enhanced expression and therapeutic benefit.

[0011] In another aspect, a vector comprises a nucleic acid sequence encoding a cytokine receptor amino acid sequence having an amino acid sequence identity of at least 75% to SEQ ID NOs: : 2, 4, 4A, 5, 5A, 6 or 6A. In certain embodiments, the vector comprises a nucleic acidsequence encoding a cytokine receptor amino acid sequence having an amino acid sequence identity of at least 95% to SEQ ID NOs: : 2, 4, 4A, 5, 5A, 6 or 6A. In certain embodiments, the vector comprises a nucleic acid sequence encoding a cytokine receptor amino acid sequence comprising SEQ ID NOs: : 2, 4, 4A, 5, 5A, 6 or 6A.

[0012] In another aspect, a pharmaceutical composition comprising a chimeric cytokine receptor embodied herein or a vector expressing a chimeric cytokine receptor embodied herein.

[0013] In another aspect, a host cell comprises a vector embodied herein. In certain embodiments, the host cell comprises an autologous cell, an allogeneic cell, a haplotype matched cell, a haplotype mismatched cell, a haplo-identical cell, a xenogeneic cell, a stem cell, cell lines or combinations thereof. In certain embodiments, the host cell comprises an immune cell. In certain embodiments, the immune cell is a natural killer (NK) cell.

[0014] In another aspect, a method of modulating natural killer (NK) cell persistence in vivo, comprises administering to a subject, a pharmaceutical composition comprising a therapeutically effective amount of a chimeric cytokine receptor, or an NK cell transduced with a vector encoding for the chimeric cytokine receptor, wherein the chimeric cytokine receptor comprises: (i) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin 15 receptor (IL-15R) domains, an IL-15R transmembrane domain, and an IL-15R intracellular domain; and (ii) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin common gamma chain (yc) domains, a yc transmembrane domain, and a yc intracellular domain. In certain embodiments, the NK cells comprise an autologous cell, an allogeneic cell, a haplotype matched cell, a haplotype mismatched cell, a haplo-identical cell, or combinations thereof. In certain embodiments, the NK cell is an autologous cell. In certain embodiments, the NK cells are transduced ex vivo with the chimeric cytokine receptor molecule. In certain embodiments, the transduced NK cells are adoptively transferred to the subject. In certain embodiments, further comprises administering one or more secondary therapies. In certain embodiments, the one or more secondary therapies comprise chemotherapeutic agents, immune therapeutic agents, checkpoint inhibitors, radiation, surgery or combinations thereof.

[0015] In another aspect, a method of treating a virus infection or cancer, comprises administering to a subject, a pharmaceutical composition comprising a therapeutically effective amount of a chimeric cytokine receptor, or an isolated cell transduced with a vector encoding for the chimeric cytokine receptor, wherein the chimeric cytokine receptor comprises: (i) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor domains, an interleukin receptor transmembrane domain, and an interleukin receptor intracellular domain; and (ii) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor common gamma chain (yc) domains, a yc transmembrane domain, and a yc intracellular domain. In certain embodiments, the isolated cell comprises an autologous cell, an allogeneic cell, a haplotype matched cell, a haplotype mismatched cell, a haplo-identical cell, or combinations thereof. In certain embodiments, the isolated cell is an autologous cell. In certain embodiments, the isolated cells are transduced ex vivo with the chimeric cytokine receptor molecule. In certain embodiments, the transduced isolated cells are adoptively transferred to the subject. In certain embodiments, treat further comprises administering one or more secondary therapies.

[0016] In another aspect, the amino acid sequences comprising SEQ ID NOs: : 2, 4, 4A, 5, 5A, 6 or 6A, comprise one or more modified amino acids, non-natural amino acids, variants or combinations thereof.

[0017] Definitions

[0018] Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that this disclosure is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary.

[0019] Standard nomenclature is used for the natural amino acids and their abbreviations. For example, L-alanine is represented with the three-letter abbreviation Ala, or one-letter abbreviation “A”. Where indicated, the “D” stereoisomer of alanine is represented as D-Ala.

[0020] Standard nomenclature is used for the bases of DNA, with cytosine, guanosine, adenine, and thymine indicated as “C”, “G”, “A”, and “T”, and codons that encode DNA follow the standard genetic code, for example the amino acid Leu is encoded by TTA, TTG, CTT, CTC, CTA or CTG, and Asp is encoded by GAT or GAC.

[0021] As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. Furthermore, to the extent that the terms “including”, “includes”, “having”, “has”, “with”, or variants thereof are used in either the detailed description and / or the claims, such terms are intended to be inclusive in a manner similar to the term “comprising.”

[0022] The term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value or range. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, within 5-fold, and also within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed. All numeric values are herein assumed to be modified by the term “about”, whether or not explicitly indicated. The recitation of numerical ranges by endpoints includes all numbers within that range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, and 5).

[0023] In the description and in the claims, phrases such as “at least one of’ or “one or more of’ may occur followed by a conjunctive list of elements or features. The term “and / or” may also occur in a list of two or more elements or features. Unless otherwise implicitly or explicitly contradicted by the context in which it is used, such a phrase is intended to mean any of the listed elements or features individually or any of the recited elements or features in combination with any of the other recited elements or features. For example, the phrases “at least one of A and B;” “one or more of A and B;” and “A and / or B” are each intended to mean “A alone, B alone, or A and B together.” A similar interpretation is also intended for lists including three or more items.For example, the phrases “at least one of A, B, and C”, “one or more of A, B, and C” and “A, B, and / or C” are each intended to mean “A alone, B alone, C alone, A and B together, A and C together, B and C together, or A and B and C together.” In addition, use of the term “based on,” above and in the claims is intended to mean, “based at least in part on,” such that an unrecited feature or element is also permissible.

[0024] As used herein, the term “affinity” is meant as a measure of binding strength. Without being bound to theory, affinity depends on the closeness of stereochemical fit between antibody combining sites and antigen determinants, on the size of the area of contact between them, and on the distribution of charged and hydrophobic groups. Affinity also includes the term “avidity,” which refers to the strength of the antigen-antibody bond after formation of reversible complexes. Methods for calculating the affinity of an antibody for an antigen are known in the art, including use of binding experiments to calculate affinity. Antibody activity in functional assays (e.g., flow cytometry assay) is also reflective of antibody affinity. Antibodies and affinities can be phenotypically characterized and compared using functional assays (e.g., flow cytometry assay).

[0025] As used herein, the term “agent” is meant to encompass any molecule, chemical entity, composition, drug, therapeutic agent, chemotherapeutic agent, or biological agent capable of preventing, ameliorating, or treating a disease or other medical condition. The term includes small molecule compounds, antisense oligonucleotides, siRNA reagents, antibodies, antibody fragments bearing epitope recognition sites, such as Fab, Fab', F(ab')z fragments, Fv fragments, single chain antibodies, antibody mimetics (such as DARPins, affibody molecules, affilins, affitins, anticalins, avimers, fynomers, Kunitz domain peptides and monobodies), peptoids, aptamers; enzymes, peptides organic or inorganic molecules, natural or synthetic compounds and the like. An agent can be assayed in accordance with the methods of the disclosure at any stage during clinical trials, during pre-trial testing, or following FDA-approval.

[0026] By “ameliorate” is meant decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease.

[0027] The term “amino acid” as used herein refers to naturally occurring and synthetic a, P, y, and 5 amino acids, and includes but is not limited to, amino acids found in proteins, i.e.glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartate, glutamate, lysine, arginine and histidine. Alternatively, the amino acid can be a derivative of alanyl, valinyl, leucinyl, isoleucinyl, prolinyl, phenylalaninyl, tryptophanyl, methioninyl, glycinyl, serinyl, threoninyl, cysteinyl, tyrosinyl, asparaginyl, glutaminyl, aspartoyl, glutaroyl, lysinyl, argininyl, histidinyl, P- alanyl, P-valinyl, P-leucinyl, P-isoleucinyl, P-prolinyl, P-phenylalaninyl, P -tryptophanyl, P- methioninyl, P-glycinyl, P-serinyl, P-threoninyl, P-cysteinyl, P-tyrosinyl, P-asparaginyl, P- glutaminyl, P-aspartoyl, P-glutaroyl, P-lysinyl, P-argininyl or P-histidinyl. The amino acids can be non-naturally occurring amino acids. Examples of non-naturally occurring amino acids include, but are not limited to, D-amino acids (i.e. an amino acid of an opposite chirality to the naturally- occurring form), N-a -methyl amino acids, C-a-methyl amino acids, P-methyl amino acids and D- or L-P-amino acids. Other non-naturally occurring amino acids include, for example, P-alanine (P- Ala), norleucine (Nle), norvaline (Nva), homoarginine (Har), 4-aminobutyric acid (y-Abu), 2- aminoisobutyric acid (Aib), 6-aminohexanoic acid (s-Ahx), ornithine (orn), sarcosine, a-amino isobutyric acid, 3 -aminopropionic acid, 2,3-diaminopropionic acid (2,3-diaP), D- or L- phenylglycine, D-(trifluoromethyl)-phenylalanine, and D-p-fluorophenylalanine. When the term amino acid is used, it is considered to be a specific and independent disclosure of each of the esters of a, P, y, and 5 glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartate, glutamate, lysine, arginine and histidine in the D and L-configurations.

[0028] The term “antibody” includes monoclonal antibodies (including full length antibodies which have an immunoglobulin Fc region), antibody compositions with polyepitopic specificity, multispecific antibodies (e.g., bispecific antibodies, diabodies, and single-chain molecules, as well as antibody fragments (e.g., Fab, F(ab')2, Fab', and Fv). The term “immunoglobulin” (Ig) is used interchangeably with “antibody” herein. The basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. An IgM antibody consists of 5 of the basic heterotetramer units along with an additional polypeptide called a J chain, and contains 10 antigen binding sites, while IgA antibodies comprise from 2-5 of the basic 4-chain units which can polymerize to formpolyvalent assemblages in combination with the J chain. In the case of IgGs, the 4-chain unit is generally about 150,000 daltons. Each L chain is linked to an H chain by one covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has at the N-terminus, a variable domain (VH) followed by three constant domains (CH) for each of the a and y chains and four CH domains for p and s isotypes. Each L chain has at the N-terminus, a variable domain (VL) followed by a constant domain at its other end. The Vtis aligned with the VH and the CL is aligned with the first constant domain of the heavy chain (CHI). Particular amino acid residues are believed to form an interface between the light chain and heavy chain variable domains. The pairing of a VH and VL together forms a single antigen-binding site. For the structure and properties of the different classes of antibodies, see e.g., Basic and Clinical Immunology, 8th Edition, Daniel P. Sties, Abba I. Ten and Tristram G. Parsolw (eds), Appleton & Lange, Norwalk, Conn., 1994, page 71 and Chapter 6. The L chain from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains (CH), immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, having heavy chains designated a, 5, s, y and p, respectively. The y and a classes are further divided into subclasses on the basis of relatively minor differences in the CH sequence and function, e.g., humans express the following subclasses: IgGl, IgG2A, IgG2B, IgG3, IgG4, IgAl and IgA2. The terms “full-length antibody,” “intact antibody” or “whole antibody” are used interchangeably to refer to an antibody in its substantially intact form, as opposed to an antibody fragment. Specifically whole antibodies include those with heavy and light chains including an Fc region. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. In some cases, the intact antibody may have one or more effector functions.

[0029] An “antibody fragment” comprises a portion of an intact antibody, preferably the antigen binding and / or the variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2 and Fv fragments; diabodies; linear antibodies (see U.S. Pat. No.5,641 ,870, Example 2; Zapata et al., Protein Eng. 8(10): 1057-1062

[1995] ); single-chain antibody molecules and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies produced two identical antigen-binding fragments, called “Fab” fragments, and a residual “Fc” fragment, a designation reflecting the ability to crystallize readily. The Fab fragment consists of an entire L chain along with the variable region domain of the H chain (VH), and the first constant domain of one heavy chain (CHI). Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site. Pepsin treatment of an antibody yields a single large F(ab')2 fragment which roughly corresponds to two disulfide linked Fab fragments having different antigen-binding activity and is still capable of cross-linking antigen. Fab' fragments differ from Fab fragments by having a few additional residues at the carboxy terminus of the C HI domain including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine residue(s) of the constant domains bear a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known. The Fc fragment comprises the carboxy -terminal portions of both H chains held together by disulfides. The effector functions of antibodies are determined by sequences in the Fc region, the region which is also recognized by Fc receptors (FcR) found on certain types of cells.

[0030] By “cancer” as used herein is meant, a disease, condition, trait, genotype or phenotype characterized by unregulated cell growth or replication as is known in the art; including colorectal cancer, as well as, for example, leukemias, e.g., acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphocytic leukemia (ALL), and chronic lymphocytic leukemia, AIDS related cancers such as Kaposi's sarcoma; breast cancers; bone cancers such as Osteosarcoma, Chondrosarcomas, Ewing's sarcoma, Fibrosarcomas, Giant cell tumors, Adamantinomas, and Chordomas; Brain cancers such as Meningiomas, Glioblastomas, Lower-Grade Astrocytomas, Oligodendrocytomas, Pituitary Tumors, Schwannomas, Primary CNS Lymphoma, and Metastatic brain cancers; cancers of the head and neck including various lymphomas such as mantle cell lymphoma, non-Hodgkin’s lymphoma, adenoma, squamous cell carcinoma, laryngeal carcinoma, gallbladder and bile duct cancers, cancers of the retina such as retinoblastoma, cancers of the esophagus, gastric cancers, multiple myeloma, ovarian cancer,uterine cancer, thyroid cancer, testicular cancer, endometrial cancer, melanoma, lung cancer, bladder cancer, prostate cancer, lung cancer (including non-small cell lung carcinoma), pancreatic cancer, sarcomas, Wilms' tumor, cervical cancer, head and neck cancer, skin cancers, nasopharyngeal carcinoma, liposarcoma, epithelial carcinoma, renal cell carcinoma, gallbladder adeno carcinoma, parotid adenocarcinoma, endometrial sarcoma, multidrug resistant cancers; and proliferative diseases and conditions, such as neovascularization associated with tumor angiogenesis, macular degeneration (e.g., wet / dry AMD), corneal neovascularization, diabetic retinopathy, neovascular glaucoma, myopic degeneration and other proliferative diseases and conditions such as restenosis and polycystic kidney disease, and other cancer or proliferative disease, condition, trait, genotype or phenotype.

[0031] The term “combination therapy”, as used herein, refers to those situations in which two or more different agents are administered in overlapping regimens so that the subject is simultaneously exposed to both agents. When used in combination therapy, two or more different agents may be administered simultaneously or separately. This administration in combination can include simultaneous administration of the two or more agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, two or more agents can be formulated together in the same dosage form and administered simultaneously. Alternatively, two or more agents can be simultaneously administered, wherein the agents are present in separate formulations. In another alternative, a first agent can be administered just followed by one or more additional agents. In the separate administration protocol, two or more agents may be administered a few minutes apart, or a few hours apart, or a few days apart.

[0032] As used herein, the transitional term “comprising,” which is synonymous with “including,” “containing,” or “characterized by,” is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. When used herein the term “comprising” can be substituted with the term “containing” or “including” or sometimes when used herein with the term “having.” By contrast, the transitional phrase “consisting of’ excludes any element, step, or ingredient not specified in the claim. The transitional phrase “consisting essentially of’ limits the scope of a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed disclosure.

[0033] As used herein, the term “common gamma c (yc) chain” or “gamma c (yc) chain” refers to the wild type or native molecule (HGNC:HGNC:6010;Ensembl:ENSG00000147168 MIM:308380; AllianceGenome:HGNC:6010; ACCESSION NP 000197).

[0034] The term “diabodies” refers to small antibody fragments prepared by constructing sFv fragments with short linkers (about 5-10) residues) between the Vu and VL domains such that inter-chain but not intra-chain pairing of the V domains is achieved, thereby resulting in a bivalent fragment, i.e., a fragment having two antigen-binding sites. Bispecific diabodies are heterodimers of two “crossover” sFv fragments in which the VH and VL domains of the two antibodies are present on different polypeptide chains. Diabodies are described in greater detail in, for example, EP 404,097; WO 93 / 1 1161 ; Hollinger et al., Proc. Natl. Acad. Set. USA 90: 6444-6448 (1993).

[0035] ‘ ‘Diagnostic” or “diagnosed” means identifying the presence or nature of a pathologic condition. Diagnostic methods differ in their sensitivity and specificity. The “sensitivity” of a diagnostic assay is the percentage of diseased individuals who test positive (percent of “true positives”). Diseased individuals not detected by the assay are “false negatives.” Subjects who are not diseased and who test negative in the assay, are termed “true negatives.” The “specificity” of a diagnostic assay is 1 minus the false positive rate, where the “false positive” rate is defined as the proportion of those without the disease who test positive. While a particular diagnostic method may not provide a definitive diagnosis of a condition, it suffices if the method provides a positive indication that aids in diagnosis.

[0036] A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal's health continues to deteriorate.

[0037] “Encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Boththe coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

[0038] The term “expression” as used herein is defined as the transcription and / or translation of a particular nucleotide sequence driven by its promoter.

[0039] As used herein, “expression vector” refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient c / .s-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e g., lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide. Examples of vectors include but are not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term includes an autonomously replicating plasmid or a virus. The term is also construed to include non-plasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, polylysine compounds, liposomes, and the like. Examples of viral vectors include, but are not limited to, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, and the like.

[0040] ‘ ‘Fv” is the minimum antibody fragment which contains a complete antigenrecognition and -binding site. This fragment consists of a dimer of one heavy- and one light-chain variable region domain in tight, non-covalent association. From the folding of these two domains emanate six hypervariable loops (3 loops each from the H and L chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three HVRs specific for an antigen) can recognize and bind antigen, although at a lower affinity than the entire binding site.

[0041] The term “hinge” or “hinge region” refers to a flexible connector region, e g. natural or synthetic polypeptides, or any other type of molecule, providing structural flexibility and spacing to flanking polypeptide regions.

[0042] ‘ ‘Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In one embodiment, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from an HVR (hereinafter defined) of the recipient are replaced by residues from an HVR of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and / or capacity. In some instances, framework (“FR”) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications may be made to further refine antibody performance, such as binding affinity. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin sequence, and all or substantially all of the FR regions are those of a human immunoglobulin sequence, although the FR regions may include one or more individual FR residue substitutions that improve antibody performance, such as binding affinity, isomerization, immunogenicity, etc. The number of these amino acid substitutions in the FR are typically no more than 6 in the H chain, and in the L chain, no more than 3. The humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See also, for example, Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1 :105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Pat. Nos. 6,982,321 and 7,087,409.

[0043] A “human antibody” is an antibody that possesses an amino-acid sequence corresponding to that of an antibody produced by a human and / or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibodyspecifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phagedisplay libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Also available for the preparation of human monoclonal antibodies are methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1 ):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e.g., immunized xenomice (see, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584 regarding XENOMOUSE™ technology). See also, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies generated via a human B-cell hybridoma technology.

[0044] The term “hypervariable region,” “HVR,” or “HV,” when used herein refers to the regions of an antibody variable domain which are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six HVRs; three in the VH (Hl, H2, H3), and three in the VL (LI, L2, L3). In native antibodies, H3 and L3 display the most diversity of the six HVRs, and H3 in particular is believed to play a unique role in conferring fine specificity to antibodies. See, e.g., Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, in Methods in Molecular Biology 248: 1-25 (Lo, ed., Human Press, Totowa, N.J., 2003). Indeed, naturally occurring camelid antibodies consisting of a heavy chain only are functional and stable in the absence of light chain. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol. 3:733-736 (1996). A number of HVR delineations are in use and are encompassed herein. The Kabat Complementarity Determining Regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). Chothia refers instead to the location of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The AbM HVRs represent a compromise between the Kabat HVRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modelingsoftware. The “contact” HVRs are based on an analysis of the available complex crystal structures.The residues from each of these HVRs are noted below.

[0045] As used herein, the term “immune cells” refers to any cells of the immune system that are involved in mediating an immune response. Non-limiting examples of immune cells include a T lymphocyte, B lymphocyte, natural killer (NK) cell, macrophage, eosinophil, mast cell, dendritic cell, neutrophil, or combination thereof. In some aspects, an immune cell expresses CD3. In certain aspects, the CD3-expressing immune cells are T cells (e.g., CD4+T cells or CD8+T cells). In some aspects, an immune cell that can be targeted with a targeting moiety (e.g., anti- CD3) comprises a naive CD4+T cell. In some aspects, an immune cell comprises a memory CD4+T cell. In some aspects, an immune cell comprises an effector CD4+T cell. In some aspects, an immune cell comprises a naive CD8+T cell. In some aspects, an immune cell comprises a memory CD8+T cell. In some aspects, an immune cell comprises an effector CD8+T cell. In some aspects, an immune cell comprises a gamma delta T cell. In some aspects, an immune cell is a dendritic cell. In certain aspects, a dendritic cell comprises a plasmacytoid dendritic cell (pDC), a conventional dendritic cell 1 (cDCl), a conventional dendritic cell 2 (cDC2), inflammatory monocyte derived dendritic cells, Langerhans cells, dermal dendritic cells, lysozyme-expressing dendritic cells (LysoDCs), Kupffer cells, or any combination thereof.

[0046] As used herein, the terms “immunoglobulin domain” and “immunoglobulin-like domain” are synonymous with “antibody” and refer to a polypeptide from the immunoglobulin superfamily of proteins with a specific secondary and tertiary structure derived from proteins found in nature. Engineered immunoglobulin domains can include components of monoclonal antibodies, antibody fragments such as Fab fragment, a F(ab')2 fragment, a single-chain variable fragment (scFv), a single-chain antibody fragment (scAb), a single domain heavy chain antibody, and a single domain light chain antibody, VHH antibodies (or nanobodies), and other single domain antibodies (sdAbs). Natural immunoglobulin and immunoglobulin-like domains are found in cell surface receptors, co-receptors, and co-activators and in soluble proteins involved in the recognition, binding and adhesion processes of cells.

[0047] The term “IL- 15 receptor molecule” or “native IL-15RP” as used herein refers to a full-length naturally-occurring IL-15 receptor 0 (IL-15RP) (e.g., a mammalian IL-15RP, e.g.,human IL-15RP, e.g., GenBank Accession Number CAG30392, Version CAG30392.1 ; embl accession CR456506.1; UniProt: P14784), an active fragment of IL-15RP, or an active variant having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to a naturally- occurring wild type polypeptide of IL-15Ra or fragment thereof. Exemplified native mammalian IL- 15 receptor beta nucleic acid or amino acid sequences can be from, for example, human, primate, canine, feline, porcine, equine, bovine, ovine, rodentia, murine, rat, hamster, guinea pig, etc. In some embodiments, the variant is a derivative, e.g., a mutant, of a wild type polypeptide or nucleic acid encoding the same. In some embodiments, the IL-15RP variant, e.g., active variant of IL-15RP, has at least 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity of the wild type IL-15RP polypeptide. In some embodiments, the IL-15RP molecule comprises one or more post-translational modifications. As used herein, the terms IL-15R and IL- 15RP are interchangeable.

[0048] The “Gammaretrovirus” is a genus in the Retroviridae family (subfamily Orthoretroviridae; Genus: Gammaretrovirus). These viruses contain a single-stranded RNA encapsulated in a nucleocapsid and enveloped by a plasma membrane derived from infected host cells during virus budding. Besides viral encoded nucleocapsid (Gag) and envelope (Env) proteins, each virion contains two identical single-stranded RNAs and enzymes including reverse transcriptase and integrase essential for virus replication. Viral cell entry is mediated through the interaction between viral Env protein and its cell surface receptor. They belong to the same family as lentiviruses, however there are a few important differences between these two genera. First, these viruses are derived from different genomes (Moloney Murine Leukemia Virus and Murine Stem Cell Virus in the case of retrovirus and the human immunodeficiency virus for lentiviruses), second and most important difference is that in contrast to lentiviruses, gammaretroviruses can transduce only dividing cells, as they lack the nuclear import mechanisms that allow entry into the nucleus of non-dividing cells. In certain embodiments, the vector comprises a gamma retrovirus.

[0049] The term “linker”, also referred to as a “spacer” or “spacer domain” as used herein, refers to an amino acid or sequence of amino acids that that is optionally located between two amino acid sequences in a fusion protein of the invention.

[0050] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and / or posttranslation modifications (e.g., isomerizations, amidations) that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In contrast to polyclonal antibody preparations which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, the monoclonal antibodies are advantageous in that they are synthesized by the hybridoma culture, uncontaminated by other immunoglobulins. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present disclosure may be made by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler and Milstein., Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14 (3): 253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nded. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567), phage-display technologies (see, e.g., Clackson et al., Nature, 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee etal., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004), and technologies for producing human or human-like antibodies in animals that have parts or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA 90: 2551 (1993); Jakobovits et al., Nature 362: 255-258 (1993); Bruggemann et al., Year in Immunol. 7:33 (1993); U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and U.S. Pat. No. 5,661,016; Marks etal., Bio / Technology 10: 779-783 (1992); Lonberg et al., Nature 368: 856-859 (1994); Morrison, Nature 368: 812-813 (1994); Fishwild et al., Nature Biotechnol. 14: 845-851 (1996); Neuberger,Nature Biotechnol. 14: 826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13: 65-93 (1995).

[0051] As used herein, a “natural amino acid” refers to the twenty genetically encoded alpha-amino acids. See, e.g., Biochemistry by L. Stryer, 3rded. 1988, Freeman and Company, New York for structures of the twenty natural amino acids.

[0052] As may be used herein, the terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid oligomer,” “oligonucleotide,” “nucleic acid sequence,” “nucleic acid fragment” and “polynucleotide” are used interchangeably and are intended to include, but are not limited to, a polymeric form of nucleotides covalently linked together that may have various lengths, either deoxyribonucleotides or ribonucleotides, or analogs, derivatives or modifications thereof. Different polynucleotides may have different three-dimensional structures, and may perform various functions, known or unknown. Non-limiting examples of polynucleotides include a gene, a gene fragment, an exon, an intron, intergenic DNA (including, without limitation, heterochromatic DNA), messenger RNA (mRNA), transfer RNA, ribosomal RNA, a ribozyme, cDNA, a recombinant polynucleotide, a branched polynucleotide, a plasmid, a vector, isolated DNA of a sequence, isolated RNA of a sequence, a nucleic acid probe, and a primer. Polynucleotides useful in the methods of the disclosure may comprise natural nucleic acid sequences and variants thereof, artificial nucleic acid sequences, or a combination of such sequences.

[0053] Unless otherwise specified, a “nucleotide sequence encoding” an amino acid sequence includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intron(s).

[0054] “Operably linked” refers to a juxtaposition wherein the components so described are in a relationship permitting them to function in their intended manner. A control sequence “operably linked” to a coding sequence is ligated in such a way that expression of the coding sequence is achieved under conditions compatible with the control sequences.

[0055] “Optional” or “optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where the event or circumstance occurs and instances where it does not

[0056] As used in this specification and the appended claims, the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise and should be understood to mean “either or both” of the elements so conjoined, e.g., elements that are conjunctively present in some cases and disjunctively present in other cases.

[0057] ‘ ‘Parenteral” administration of an immunogenic composition includes, e g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), intravitreal (i.v.i.), intra-cisterna magna (i.c.m.), or intrasternal injection, or infusion techniques.

[0058] The terms “patient” or “individual” or “subject” are used interchangeably herein, and refers to a mammalian subject to be treated, with human patients being preferred. In some cases, the methods of the invention find use in experimental animals, in veterinary application, and in the development of animal models for disease, including, but not limited to, rodents including mice, rats, and hamsters, and primates.

[0059] “Percentage of sequence identity” is determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. In embodiments, the percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.

[0060] The terms “pharmaceutically acceptable” (or “pharmacologically acceptable”) refer to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to an animal or a human, as appropriate. The term “pharmaceutically acceptable carrier,” as used herein, includes any and all solvents, dispersionmedia, coatings, antibacterial, isotonic and absorption delaying agents, buffers, excipients, binders, lubricants, gels, surfactants and the like, that may be used as media for a pharmaceutically acceptable substance.

[0061] A polynucleotide is typically composed of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T) (uracil (U) for thymine (T) when the polynucleotide is RNA). Thus, the term “polynucleotide sequence” is the alphabetical representation of a polynucleotide molecule; alternatively, the term may be applied to the polynucleotide molecule itself. This alphabetical representation can be input into databases in a computer having a central processing unit and used for bioinformatics applications such as functional genomics and homology searching. Polynucleotides may optionally include one or more non-standard nucleotide(s), nucleotide analog(s) and / or modified nucleotides.

[0062] The terms “polypeptide,” “peptide” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues, wherein the polymer may in embodiments be conjugated to a moiety that does not consist of amino acids. The terms also apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. A “fusion protein” refers to a chimeric protein encoding two or more separate protein sequences that are recombinantly expressed or chemically synthesized as a single moiety.

[0063] “Polypeptide fragment” refers to a polypeptide that has an amino-terminal and / or carboxy-terminal deletion, in which the remaining amino acid sequence is usually identical to the corresponding positions in the naturally-occurring sequence. Fragments typically are at least 5, 6, 8 or 10 amino acids long, at least 14 amino acids long, at least 20 amino acids long, at least 50 amino acids long, or at least 70 amino acids long.

[0064] The term “promoter” as used herein is defined as a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence.

[0065] As used herein, the term “promoter / regulatory sequence” means a nucleic acid sequence which is required for expression of a gene product operably linked to the promoter / regulatory sequence. In some instances, this sequence may be the core promoter sequence and in other instances, this sequence may also include an enhancer sequence and other regulatory elements which are required for expression of the gene product. The promoter / regulatory sequence may, for example, be one which expresses the gene product in a tissue specific manner.

[0066] A “constitutive” promoter is a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell.

[0067] An “inducible” promoter is a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell substantially only when an inducer which corresponds to the promoter is present in the cell.

[0068] A “tissue-specific” promoter is a nucleotide sequence which, when operably linked with a polynucleotide encodes or specified by a gene, causes the gene product to be produced in a cell substantially only if the cell is a cell of the tissue type corresponding to the promoter.

[0069] A “therapeutic” treatment is a treatment administered to a subject who exhibits signs of pathology, for the purpose of diminishing or eliminating those signs.

[0070] The term “transfected” or “transformed” or “transduced” means to a process by which exogenous nucleic acid is transferred or introduced into the host cell. A “transfected” or “transformed” or “transduced” cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The transfected / transformed / transduced cell includes the primary subject cell and its progeny.

[0071] “Single-chain Fv” also abbreviated as “sFv” or “scFv” are antibody fragments that comprise the VH and VL antibody domains connected into a single polypeptide chain. Preferably, the sFv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the sFv to form the desired structure for antigen binding. For a review of the sFv, seePluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994). “Functional fragments” of the antibodies of the disclosure comprise a portion of an intact antibody, generally including the antigen binding or variable region of the intact antibody or the Fc region of an antibody which retains or has modified FcR binding capability. Examples of antibody fragments include linear antibody, single-chain antibody molecules and multispecific antibodies formed from antibody fragments.[00072J As used herein, an “unnatural amino acid,” “non-natural”, “modified amino acid” or “chemically modified amino acid” refers to any amino acid, modified amino acid, or amino acid analogue other than the twenty genetically encoded alpha-amino acids. Unnatural amino acids have side chain groups that distinguish them from the natural amino acids, although unnatural amino acids can be naturally occurring compounds other than the twenty proteinogenic alphaamino acids. In addition to side chain groups that distinguish them from the natural amino acids, unnatural amino acids may have an extended backbone such as beta-amino acids.

[0073] Non-limiting examples of non-natural amino acids include selenocysteine, pyrrolysine, homocysteine, an O-methyl-L-tyrosine, an L-3-(2-naphthyl)alanine, a 3-methyl- phenylalanine, an O-4-allyl-L-tyrosine, a 4-propyl-L-tyrosine, a tri-O-acetyl-GlcNAcP-serine, an L-Dopa, a fluorinated phenylalanine, an isopropyl-L-phenylalanine, a p-azido-L -phenylalanine, a p-acyl-L-phenylalanine, a p-benzoyl-L-phenylalanine, an L-phosphoserine, a phosphonoserine, a phosphonotyrosine, a p-iodo-phenylalanine, a p-bromophenylalanine, a p-amino-L-phenylalanine, an isopropyl-L-phenylalanine, an unnatural analogue of a tyrosine amino acid; an unnatural analogue of a glutamine amino acid; an unnatural analogue of a phenylalanine amino acid; an unnatural analogue of a serine amino acid; an unnatural analogue of a threonine amino acid; an alkyl, aryl, acyl, azido, cyano, halo, hydrazine, hydrazide, hydroxyl, alkenyl, alkynl, ether, thiol, sulfonyl, seleno, ester, thioacid, borate, boronate, phospho, phosphono, phosphine, heterocyclic, enone, imine, aldehyde, hydroxylamine, keto, or amino substituted amino acid, or any combination thereof; an amino acid with a photoactivatable cross-linker; a spin-labeled amino acid; a fluorescent amino acid; an amino acid with a novel functional group; an amino acid that covalently or noncovalently interacts with another molecule; a metal binding amino acid; a metal-containing amino acid; a radioactive amino acid; a photocaged and / or photoisomerizable amino acid; a biotinor biotin-analogue containing amino acid; a glycosylated or carbohydrate modified amino acid; a keto containing amino acid; amino acids comprising polyethylene glycol or polyether; a heavy atom substituted amino acid; a chemically cleavable or photocleavable amino acid; an amino acid with an elongated side chain; an amino acid containing a toxic group; a sugar substituted amino acid, e.g., a sugar substituted serine or the like; a carbon-linked sugar-containing amino acid; a redox-active amino acid; an a-hydroxy containing acid; an amino thio acid containing amino acid; an a, a disubstituted amino acid; a P-amino acid; and a cyclic amino acid other than proline. In an embodiment of the helicases described herein, one or more amino acids of the helicase are substituted with one or more unnatural amino acids and / or one or more natural amino acids.

[0074] The “variable region” or “variable domain” of an antibody refers to the aminoterminal domains of the heavy or light chain of the antibody. The variable domains of the heavy chain and light chain may be referred to as “VH” and “VL”, respectively. These domains are generally the most variable parts of the antibody (relative to other antibodies of the same class) and contain the antigen binding sites. However, the variability is not evenly distributed across the entire span of the variable domains. Instead, it is concentrated in three segments called hypervariable regions (HVRs) both in the light-chain and the heavy chain variable domains. The more highly conserved portions of variable domains are called the framework regions (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three HVRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The HVRs in each chain are held together in close proximity by the FR regions and, with the HVRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991)). The constant domains are not involved directly in the binding of antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.

[0075] As used herein, “variant” of polypeptides refers to an amino acid sequence that is altered by one or more amino acid residues. The variant may have “conservative” changes, wherein a substituted amino acid has similar structural or chemical properties (e.g., replacement of leucine with isoleucine). More rarely, a variant may have “nonconservative” changes (e.g., replacementof glycine with tryptophan). Analogous minor variations may also include amino acid deletions or insertions, or both. Guidance in determining which amino acid residues may be substituted, inserted, or deleted without abolishing biological activity may be found using computer programs well known in the art, for example, LASERGENE software (DNASTAR).

[0076] “ Treatment” is an intervention performed with the intention of preventing the development or altering the pathology or symptoms of a disorder. Accordingly, “treatment” refers to both therapeutic treatment and prophylactic or preventative measures. “Treatment” may also be specified as palliative care. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented. Accordingly, “treating” or “treatment” of a state, disorder or condition includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in a human or other mammal that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition; (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical or subclinical symptom thereof; or (3) relieving the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms. The benefit to an individual to be treated is either statistically significant or at least perceptible to the patient or to the physician.

[0077] ‘ ‘Variant” as the term is used herein, is a nucleic acid sequence or a peptide sequence that differs in sequence from a reference nucleic acid sequence or peptide sequence respectively, but retains essential properties of the reference molecule. Changes in the sequence of a nucleic acid variant may not alter the amino acid sequence of a peptide encoded by the reference nucleic acid, or may result in amino acid substitutions, additions, deletions, fusions and truncations. Changes in the sequence of peptide variants are typically limited or conservative, so that the sequences of the reference peptide and the variant are closely similar overall and, in many regions, identical. A variant and reference peptide can differ in amino acid sequence by one or more substitutions, additions, deletions in any combination. A variant of a nucleic acid or peptide can be a naturally occurring such as an allelic variant, or can be a variant that is not known to occurnaturally. Non-naturally occurring variants of nucleic acids and peptides may be made by mutagenesis techniques or by direct synthesis.

[0078] As used herein, the term “virus” includes any type of virus or virus vector. For example, adenovirus, adeno-associated virus (AAV), recombinant adeno-associated virus (rAAV), herpes simplex virus, lentivirus, retrovirus, alphavirus, flavivirus, rhabdovirus, measles virus, Newcastle disease virus, poxvirus, vaccinia virus, modified Ankara virus, vesicular stomatitis virus, picornavirus. In various embodiments the virus is a chimeric virus, a synthetic virus, a recombinant virus, a mosaic virus or a pseudotyped virus.

[0079] Ranges: throughout this disclosure, various aspects of the disclosure can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range. The recitation of numerical ranges by endpoints includes all numbers, e.g., whole integers, including fractions thereof, subsumed within that range (for example, the recitation of 1 to 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, and the like) and any range within that range.

[0080] All genes, gene names, and gene products disclosed herein are intended to correspond to homologs from any species for which the compositions and methods disclosed herein are applicable. Thus, the terms include, but are not limited to genes and gene products from humans and mice. It is understood that when a gene or gene product from a particular species is disclosed, this disclosure is intended to be exemplary only, and is not to be interpreted as a limitation unless the context in which it appears clearly indicates. Thus, for example, for the genes or gene products disclosed herein, which in some embodiments relate to mammalian nucleic acid and amino acid sequences, are intended to encompass homologous and / or orthologous genes and gene products from other animals including, but not limited to other mammals, fish, amphibians,reptiles, and birds. In preferred embodiments, the genes, nucleic acid sequences, amino acid sequences, peptides, polypeptides and proteins are human. The term “gene” is also intended to include variants.

[0081] The practice of the present disclosure employs, unless otherwise indicated, conventional techniques of chemistry, molecular biology, microbiology, recombinant DNA, genetics, immunology, cell biology, cell culture and transgenic biology, which are within the skill of the art. See, e.g., Maniatis et al., 1982, Molecular Cloning (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Sambrook etal., 1989, Molecular Cloning, 2nd Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Sambrook and Russell, 2001, Molecular Cloning, 3rd Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Ausubel et al., 1992), Current Protocols in Molecular Biology (John Wiley & Sons, including periodic updates); Glover, 1985, DNA Cloning (IRL Press, Oxford); Anand, 1992; Guthrie and Fink, 1991; Harlow and Lane, 1988, Antibodies, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.); Jakoby and Pastan, 1979; Nucleic Acid Hybridization (B. D. Hames & S. J. Higgins eds. 1984); Transcription And Translation (B. D. Hames & S. J. Higgins eds. 1984); Culture Of Animal Cells (R. I. Freshney, Alan R. Liss, Inc., 1987); Immobilized Cells And Enzymes (IRL Press, 1986); B. Perbal, A Practical Guide To Molecular Cloning (1984); the treatise, Methods In Enzymology (Academic Press, Inc., N.Y.); Gene Transfer Vectors For Mammalian Cells (J. H. Miller and M. P. Calos eds., 1987, Cold Spring Harbor Laboratory); Methods In Enzymology, Vols. 154 and 155 (Wu et al. eds.), Immunochemical Methods In Cell And Molecular Biology (Mayer and Walker, eds., Academic Press, London, 1987); Handbook Of Experimental Immunology, Volumes I-IV (D. M. Weir and C. C. Blackwell, eds., 1986); Riott, Essential Immunology, 6th Edition, Blackwell Scientific Publications, Oxford, 1988; Hogan et al., Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986); Westerfield, M., The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerid), (4th Ed., Univ, of Oregon Press, Eugene, 2000).

[0082] Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0083] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and payment of the necessary fee.

[0084] FIGS. 1A-1D are a series of schematics showing the chimeric cytokine receptor design based on the interleukin- 15 receptor (IL-15R). FIG. 1A: Endogenous interaction of heterodimeric common cytokine receptor gamma chain family receptors involves interaction of the common gamma chain (yc) and a receptor chain that confers specificity to a cytokine. FIG. IB: Validated single chain variable fragments (scFvs) targeting CD123 (26292 and 32716) were added to various sequences derived from the yc and IL-15RP chains to create chimeric cytokine receptors (CCRs). Binding a cell surface antigen on a target cell will pull the heterodimeric CCRs together and induce signal transduction by the Janus kinase / signal transducer and activator of transcription (JAK / STAT) pathway. FIG. 1C: AlphaFold was used to model the folding of the chimeric yc receptors (CCR-yc) with shortening segments of the native extracellular yc, all including the same scFv (32716). Red and blue constructs were previous attempts from our group at creating a CCR-yc but constructs either did not express or did not bind the target antigen. FIG. ID: From in silica predictions, two CCR-yc (pink and white in FIG. 1C) were chosen for in vitro testing. Three CCRs using IL-15RP sequences (CCR-IL- 15RP) were designed using similar principles, all including the 26292 scFv. These constructs were subcloned into pSFG expression vectors containing fluorescent proteins for subcellular tracking (green fluorescent protein (GFP) or a red fluorescent protein (mScarlet, mSc)) and were designated 15D2a.mSc, 15D2.mSc, 15Dl.mSc, ycD2.GFP, and ycDl.GFP. SnapGene v5.1.7 software was used to design subcloning procedures. Plasmid sequence fidelity was validated by Sanger sequencing (Johns Hopkins Genomic Research Core Facilities).

[0085] FIG. 2 is series of plots demonstrating CCR expression and antigen binding is dependent on the length of the native extracellular domain included. Primary NK cells derived from healthy donor peripheral blood were transduced with BaeV-pseudotyped replicationincompetent gamma-retroviral vectors containing our chimeric yc and chimeric IL-15RP of different lengths. Transduction efficiency was assessed by flow cytometry 2 to 4 days afterincubation with virus. The longest CCR-yc and CCR-IL-15RP (15Dl .mSc and ycDl .GFP, respectively) had the highest expression as detected by fluorescent protein expression. Higher antigen binding was also observed with these constructs after incubation with his-tagged recombinant human CD123 (rhCD123) for 30 minutes and staining with anti-his-tag flow antibodies.

[0086] FIGS. 3A-3D are a series of schematics and plots demonstrating CCR-yc and CCR- IL-15R0 exhibit stable surface expression and antigen binding individually or together. FIG. 3A: Sequence diagrams of CCR-pair with the highest expression and binding. FIG. 3B: The total expression, antigen binding, and surface expression of this CCR-pair were assessed using flow cytometry. Total expression was calculated as the percentage of cells positive for GFP and / or mScarlet (GFP+and / or mSc+). Antigen binding was determined after incubation with his-tagged rhCD123 for 30 minutes and staining with anti-his-tag flow antibodies (His+). Surface expression was determined by staining with an Alexa Fluor 647-conjugated anti-mouse IgG flow antibody (F(ab)2+), specific for murine antibody variable regions (i.e., able to bind an scFv independent of antigen). Compensation and marker positivity was calculated using FlowJo vlO.lO.O software. N = 3 unique NK cell donors. FIG. 3C: Representative dot plots of NK cells expressing each CCR chain individually and together. FIG. 3D shows SEQ ID NOS. 1-4 including specified regions thereof. FIG. 3D is the specified SEQ ID NOs: 1 and 2.

[0087] FIGS. 4A-4D CCRs induce antigen-specific signaling and activation in response to stimulation with CD 123 and includes a series of graphs and a blot demonstrating that CCRs induce antigen-specific signaling and activation in response to stimulation with CD 123. FIG. 4A: Non-transduced, 15Dl.mSc, ycDl.GFP, and co-transduced NK cells were stimulated with platebound rhCD123 or rhIL-2 (lane 3, positive control for antibody) for 30 minutes. Unstimulated cells served as negative controls. Cells were lysed, total protein quantified by BCA, and then western blot was performed on 20 pg of total protein from each lysate. STAT5 is phosphorylated (pSTAT5) downstream of yc cytokine family receptors. Antibody stains were used for total STAT5 and pSTAT5 with a GAPDH loading control. Lanes were quantified in imageJ, normalized to GAPDH, and the ratio of pSTAT5 Total STAT5 was calculated. Co-transduced NK cells showed a 2-fold increase in signal when stimulated with rhCD123. N = 3 NK cell donors. FIG. 4B: To assess iffunctional signaling was necessary for CCR functionality in NK cells, mutated ycDl GFP constructs were created using site-directed mutagenesis. Two signaling null mutants were created by introducing an early stop codon after the transmembrane region (R289*, mutation of residue corresponding to R289 in the native yc) or by deleting the JAK binding motif (Boxl A, deletion of residues corresponding to T286 through K294 in the native yc). All CCR-yc constructs showed similar levels of transduction in NK cells as detected by flow cytometry using his-tagged rhCD123 and anti-his-tag antibody staining. To assess if functional scFv-binding was necessary for CCR functionality in NK cells, a native yc fused to GFP (yc-GFP) was cloned into an pSFG expression vector. Expression of yc-GFP was achieved in NK cells based on detection of GFP signal by flow cytometry. Representative flow plots. FIG. 4C: NK cells were transduced with mutated chimeric ycDl.GFPs (R289* or BoxlA), yc-GFP, and chimeric ycDl.GFP alone and in combination with our best expressing CCR-IL-15RP (15Dl.mSc). Engineered NK cells were co-cultured at a 1 : 1 effector-to-target ratio (E:T, where NK cells are effector cells and target cells are cancer cell lines) with firefly luciferase (ffLuc) expressing MOLM-13 for 18 hours. Reduction in bioluminescent signal was detected and compared to signal from target cells incubated without NK cells. Only NK cells co-transduced with CCR-IL-15RP and CCR-ycDl (ycDl.GFP with intact signaling and antigen-binding) were able to activate NK cells vs. MOLM-13 and induce cytotoxicity. N = 1, 3 technical replicates per condition. FIG. 4D: NK cells engineered with functional ycDl.GFP and 15Dl.mSc, alone or together, were co-cultured with ffLuc target cells, either MOLM-13 (CD123- positive) or Raji (CD 123 -negative), for 18 hours at various E:T ratios. Cytotoxicity was calculated as a reduction in bioluminescent signal compared to signal from cancer cells incubated without NK cells. Only NK cells co-transduced with both halves of our CCR-pair displayed elevated cytotoxicity. Statistical comparison was completed using two-way ANOVA with Tukey tests for multiple comparisons: **** p < 0.0001 vs. non-transduced, N = 3 NK cell donors, 3 technical replicates per donor.[00088J FIGS. 5A-5C: High expression of both CCR chains induces robust cytotoxic activity against cancer cell lines. FIG. 5A: NK cells were engineered with monocistronic CCR- yc (ycDl), monocistronic CCR-IL-15RP (15D1), or bicistronic CCRs (ycDl.T2A.15Dl) encoded in pMSGV expression vectors. CCR chains expressed individually or together displayed long-termsurface expression and antigen binding up to 22 days post-activation, as determined by flow cytometry using his-tagged rhCD123 and anti-his-tag antibody staining. N = 3 NK cell donors. FIG 5B: CRISPR was used to knockout (KO) the target antigen of interest (CD123) from three AML cell lines (MOLM-13, MV-4-11, and 0CI-AML3) to create negative controls. KO was confirmed using flow cytometry. FIG. 5C: NK cells engineered with monocistronic OCR chains or bicistronic OCRs were co-cultured with ffLuc-expressing target cells, either CD 123 -positive targets (top: MOLM-13, MV411, 0CLAML3, Raji engineered with constitutive CD123 expression) or CD 123 -negative targets (bottom: CRISPR knockout cell lines or parental Raji), for 18 hours at various E:T ratios. Reduction in bioluminescent signal was detected and compared to signal from target cells incubated without NK cells. NK cells expressing both halves of our CCR- pair on one expression vector showed elevated cytotoxicity against all evaluated cancer cell lines. Statistical comparison was completed using two-way ANOVA with Tukey tests for multiple comparisons: * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001, color of asterisks corresponds to experimental condition vs. non-transduced, N = 3 NK cell donors, 3 technical replicates per donor.

[0089] FIGS. 6A-6D. CCR-NK cells have increased survival and sustained killing activity in vitro. FIG. 6A: NK cells were transduced with a BaeV-pseudotyped vector encoding a nuclear- localized (NLS) mScarlet alone (mSc) or with a 1 : 1 mixture ofNLS-mSc vector and our bicistronic CCR vector (ycDl.T2A.15D1, CCR+mSc). Expression of mSc and CCRs were confirmed using flow cytometry and recombinant protein staining. FIG. 6B NLS-mScarlet enabled long-term tracking of NK cells in an Incucyte S3 Live-Cell Analysis System. Engineered NK cells expressing CCRs or not were tracked over the course of 10 days with or without supplemental rhIL-2 (25 lU / mL, top). Media was refreshed every 2 days. CCR-NK cells displayed improved cytokineindependent survival but were still responsive to exogenous stimulation with cytokine. NK-cell numbers were quantified as the number of red objects normalized to the starting value at time 0. Solid bold lines represent mean NK cell counts, while thin and dashed lines are individual replicates. NK cells were also serially stimulated with NLS-GFP expressing MOLM-13 and MOLM-13.CD123KO target cells (bottom). NK cells and AML targets were plated at an initial density of 2e5: l e5 and le5 additional target cells were added every 2 days. CCR-NK cells wereable to kill AML across consecutive stimulations. CD 123+ parental MOLM-13 had a shorter halflive than MOLM-13.CD123KO co-cultured with CCR-NK cells for all stimulations except the last. Quantity of AML was calculated as the number of green objects and normalized to the number of AML cells present at the start of each repeat stimulation. Cell numbers were quantified using the Incucyte 2022B Rev2 software. AML count curve comparisons were done by fitting one-phase decay curves to each stimulation using GraphPad Prism 10. Previously collected data using chimeric antigen receptor (CAR) modified NK cells has been included for comparison. N = 3 NK cell donors, 3 technical replicates per donor, 4 images per replicate. FIG. 6C: Representative images and respective image masks (counted objects) of engineered NK cells cultured with and without rhIL-2 on day 5 of 10-day culture. FIG. 6D: Following 10-day culture with and without rhIL-2, NK cells were harvested for flow cytometry. CCR+ cells, as determined by recombinant antigen staining (His+), had a trend of enrichment without cytokine supplementation but were not significantly elevated. Statistical comparison was completed using one-way ANOVA with Tukey tests for multiple comparisons. N = 3 NK cell donors

[0090] FIGS. 7A-7D: CCR-NK cells have increased survival in vivo. FIG. 7A: The longevity of CCR-NK cells was evaluated in 8-week-old female NSG (NOD. Cg- Prkdcsc,dIl2r^mlW]lISz]') mice. Mice were first engrafted with le6 MV-4-11 via tail vein injection. Mice were treated with saline, NK cells without a chimeric construct, or CCR-NK cells 7 days following AML injections with N = 4 or 5 mice per group. NK cells were modified to express firefly luciferase (ffLuc) to enable tracking in live animals. CCR+ffLuc.NK cells were transduced with separate viral vectors on subsequent days. FIG. 7B: NK cell abundance was detected by bioluminescence imaging using an Xenogen IVIS Spectrum in vivo imaging system following intraperitoneal injection of D-luciferin. Radiance was quantified using the Living Image software. Thick lines represent means while thin dashed lines represent radiance signal for individual animals. FIG. 7C: Images collected for mice treated with ffLuc.NK and CCR+ffLuc.NK cells. FIG. 7D: One month post-treatment, all mice were sacrificed. Spleens, bone marrow (BM) from hind limbs, and peripheral blood (PB) was collected and processed for analysis by flow cytometry. NK cell abundance (top) and AML burden (bottom) was determined for each compartment.

[0091] FIG. 8A-8C: CCRs can be functionalized against another cancer-associated antigen: B7H3. FIG. 8A: NK cells were engineered with monocistronic anti-B7H3 CCR-yc, monocistronic anti-B7H3 CCR-IL-15R0, or bicistronic CCRs encoded in pMSGV expression vectors. CCR chains could be expressed individually or together, as determined by flow cytometry on day 8 post NK-cell activation using his-tagged rhB7H3 and anti-his-tag antibody staining. N = 2 NK cell donors. FIG. 8B: CRISPR was used to knockout (KO) the second target antigen of interest (B7H3) from two AML cell lines (MV-4-11 and OCLAML3) to create negative controls. KO was confirmed using flow cytometry. FIG. 8C: NK cells engineered with anti-B7H3 CCR chains were co-cultured with either B7H3 -positive or B7H3-negative ffLuc-expressing target cells for 18 hours at various E:T ratios. Reduction in bioluminescent signal was detected and compared to signal from target cells incubated without NK cells. CCR-NK cells expressing both chimeric anti-B7H3 chains showed elevated cytotoxicity against the evaluated cancer cell lines. Statistical comparison was completed using two-way ANOVA with Tukey tests for multiple comparisons: * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001, color of asterisks corresponds to experimental condition vs. non-transduced, N = 2 NK cell donors, 3 technical replicates per donor.DETAILED DESCRIPTION

[0092] In order to increase NK cell persistence, antigen-specific chimeric cytokine receptors (CCRs) based on the interleukin- 15 receptor (IL-15R) have been engineered to stimulate prolonged INK cell survival.

[0093] IL-15 / IL-15RP and Immune Cells

[0094] The cytokine interleukin (IL)-15 is crucial for natural killer (NK) cell development, survival, and proliferation. The IL-15 receptor (IL-15R) consists of 3 subunits: the a chain, unique to IL-15R; the P chain, shared with IL-2R; and the common y (yc) chain, shared with the receptors for IL-2, IL-4, IL-7, IL-9, and IL-21. Accordingly, NK cells do not develop in mice lacking either IL- 15 or any of the 3 IL-15R subunits. Despite their similarity, IL-15R has properties that are not shared with IL-2R. IL-15Ra has a very high affinity for IL- 15 (Kd= 50 pM), whereas IL-2Ra binds IL-2 with a Ala of 10 nM. IL-15Ra and IL- 15 are coexpressed in the same cells and exported to the plasma membrane as a complex, with IL-15Ra acting as a chaperone for IL-15. Therefore, IL- 15is usually presented in trans by cells expressing the IL-15Ra-IL-l 5 complex, such as stromal cells and dendritic cells (DCs), to cells that express the P-ycchains, such as T cells and NK cells. Consistent with this, adoptively transferred NK cells lacking the IL-15Ra gene survive in normal mice but not in IL-15Ra-deficient mice. Signal transduction from the [Lye chains activates multiple pathways, including a pathway that leads to a transcriptional response and cell survival through JAKl / 3-dependent phosphorylation of STAT5 and another pathway that activates cell proliferation through PI-3K-Akt-mTOR-dependent phosphorylation of the ribosomal protein S6 by the p70-S6 kinase (Anton, O.M. et al., (2019) Zra / zs-endocytosis of intact IL-15Ra-IL-15 complex from presenting cells into NK cells favors signaling for proliferation, PNAS 117 (1) 522- 531; doi.org / 10.1073 / pnas.1911678117).

[0095] Natural Killer Cells: Natural killer (NK) cells are an essential part of tumor immunosurveillance, evidenced by higher cancer susceptibility and metastasis in association with diminished NK activity in mouse models and clinical studies. Using an array of germline-encoded surface receptors, NK cells are able to recognize and rapidly act against malignant cells without prior sensitization. Upon activation, NK cells release cytotoxic granules containing perforin and granzymes to directly lyse tumor cells, in a similar fashion to activated cytotoxic T cells. NK cells are also potent producers of chemokines and cytokines such as interferon gamma (IFN-y) and tumor necrosis factor alpha (TNF-a) and thereby are essential in modulating adaptive immune responses (Liu, S., Galat, V., Galat, Y. et al. NK cell-based cancer immunotherapy: from basic biology to clinical development. J Hematol Oncol 14, 7 (2021). doi.org / 10.1186 / sl3045-020- 01014-w).

[0096] NK cells can trigger target cell death by releasing cytotoxic granules containing granzymes and perforin and through death receptor-mediated pathways (e.g., FasL / Fas) (Smyth MJ, et al. Activation of NK cell cytotoxicity. Mol Immunol. 2005;42(4):501-10). NK cells also play immunomodulatory functions by secreting chemokines and cytokines, such as RANTES and IFN-y (Roda JM, et al. Natural killer cells produce T cell-recruiting chemokines in response to antibody-coated tumor cells. Cancer Res. 2006;66(l):517-26; Bottcher JP, et al. NK cells stimulate recruitment of cDCl into the tumor microenvironment promoting cancer immune control. Cell. 2018; 172(5): 1022-1037).

[0097] In humans, NK cells are traditionally identified by the absence of CD3 and the presence of CD56 on their surface as characterized by flow cytometry. In mouse, the lack of CD3 and the presence of NK1.1 are canonical criterion for distinguishing NK cells. In mouse strains lacking NK1.1 expression (e.g., BALB / c), CD49b is used for NK cell identification. The natural cytotoxic receptor NKp46 is also often used to identify mouse and human NK cells in combination with the absence of CD3 expression. Notably, certain tissues such as the mucosal barriers possess subsets of recently identified innate lymphoid cells (ILCs) that also share canonical markers of NK cells. For example, a subset of IL-22 secreting human ILC3s is CD56 NI<p46 CD3 (Spits H, et al. Innate lymphoid cells: a proposal for uniform nomenclature. Nat Rev Immunol. 2013 ; 13(2): 145-9). Additional markers such as lack of c-kit can be used to distinguish human NK (c-kit ) from ILC3s (c-kit+). In mouse, NK and ILC1 are NK I . I CD3 but can be further characterized by CD49a and Eomes expression. NK cells are CD49a Eomes , whereas ILCls are CD49a Eomes (Gao Y, et al. Tumor immunoevasion by the conversion of effector NK cells into type 1 innate lymphoid cells. Nat Immunol. 2017;18(9): 1004-15; Colonna M. Innate lymphoid cells: diversity, plasticity, and unique functions in immunity. Immunity. 2018;48(6): 1104-17).

[0098] NK cells are found both in blood at levels of 5%-l 5% of circulating lymphocytes and in various lymphoid and non-lymphoid organs such as the spleen, lung, and liver (Freud AG, et al. The broad spectrum of human natural killer cell diversity. Immunity. 2017;47(5):820— 33; Bjorkstrom NK, et al. Emerging insights into natural killer cells in human peripheral tissues. Nat Rev Immunol. 2016; 16(5):310-20). Based on characterization ofNK cells in the peripheral blood, human NK cells are conventionally sub-divided into two major subsets: CD56briglltCD I 6dimand CD56dimCD16+, with the former classically believed to be less mature and a potent cytokine producer and the latter more mature and the most cytotoxic (Abel AM, et al. Natural killer cells: development, maturation, and clinical utilization. Front Immunol. 2018;9: 1869). Most NK cells in the blood are CD56dim, whereas the CD56brightsubset only represents less than 15% of total circulating NK cells. The relative proportion of CD56bnghtand CD56dnnNK cells in tissues can be very different from that observed in the peripheral blood. Notably, many tissue-resident NK subsets are now shown to be phenotypically and functionally distinct from conventional peripheral blood NK cells (Hashemi E, etal. Tissue-resident NK cells: development, maturation, and clinicalrelevance. Cancers (Basel), 2020; 12(6). For example, uterine NK cells, which constitute the majority of lymphocyte in the uterus during the first trimester, are CD56super bnghtand play important roles in pregnancies by regulating placental vascular remodeling (Liu, S., et al. NK cellbased cancer immunotherapy: from basic biology to clinical development. J Hematol Oncol 14, 7 (2021). doi.org / 10.1186 / sl3045-020-01014-w).

[0099] NK cells primarily develop in the bone marrow (BM), which contains abundant hematopoietic stem cells (HSCs) capable of differentiating toward NK cells through common lymphoid progenitor (CLP) and lineage-restricted progenitor (NKP) cells (Di Santo JP. Natural killer cell developmental pathways: A Question of Balance. Annu Rev Immunol (2006) 24(1):257— 86. doi: 10.1 146 / annurev.immunol.24.021605.090700). Multiple internal pathways and external factors contribute to the development of NK cells from HSCs (Stabile H, et al. Impact of bone marrow-derived signals on NK cell development and functional maturation. Cytokine Growth Factor Rev (2018) 42: 13-9. doi: 10.1016 / j.cytogfr.2018.03.008). Most importantly, the pleiotropic cytokine IL- 15 is indispensable for the development and homeostasis of NK cells as highlighted by their significant deficiency in IL-15-deficient mice. Correspondingly, deficiency in IL- 15 or any one of the IL- 15 receptor subunits, such as the IL-15Ra, IL-15RP, and yc in mice, results in a dramatic paucity of mature NK cells (Vosshenrich CA, et al. Roles for common cytokine receptor gamma-chain-dependent cytokines in the generation, differentiation, and maturation of NK cell precursors and peripheral NK cells in vivo. J Immunol (2005) 174(3): 1213-21. doi: 10.4049 / jimmunol.174.3.1213). Parallel with the role of IL-15 in mice, several studies have demonstrated that the early commitment of NK cells from human CD34+hemopoietic progenitor cells into NKP cells is dependent on the coordinated function of IL-3, IL-7, c-kit ligand (KL), and flt3 ligand (FL) but not IL-15, whereas IL- 15 is involved in the emergence of CD56+NK cells.[000100] NK cell development primarily involves the following two independent and continuous processes: early NK cell commitment to IL- 15 -responsive NK cell progenitors (NKPs) and subsequent phenotypical and functional maturation of NK cells in response to IL-15. Early NK cell commitment to NKP cells is characterized by the acquisition of CD122 (IL-15R0), which is a critical subunit of the IL-15 receptor and dimerizes with yc to transduce IL-15 signaling. However, IL-15 is not involved in the generation of IL- 15 -responsive NKPs because the IL-15receptor is not expressed prior to the NKP stage (Rosmaraki EE, et al. Identification of committed NK cell progenitors in adult murine bone marrow. Eur J Immunol (2001) 31(6): 1900-9. doi: 10.1002 / 1521-4141(200106)31:6<1900::aid-immul900>3.0.co;2-m). Pre-NKP cells were identified as the earliest committed NK cell progenitors in murine BM, and these cells reside downstream of CLP and differentiate into NKPs (Fathman JW, et al. Identification of the earliest natural killer cell-committed progenitor in murine bone marrow. Blood (2011) 118(20):5439. doi: 10.1182 / blood-2011-04-348912). Although pre-NKP cells express undetectable levels of CD122, they are fully committed to the NK lineage both in vitro and in vivo. Therefore, IL- 15 is not necessary for NK cell lineage commitment. Furthermore, mice deficient in yc exhibit an intact NKP compartment (Vosshenrich CA, et al. (2005)), and IL-3, IL-7, KL, and FL synergistically drive the differentiation of NKP cells from human HSCs in vitro in the absence of IL- 15 (McCullar V, et al. Mouse fetal and embryonic liver cells differentiate human umbilical cord blood progenitors into CD56-negative natural killer cell precursors in the absence of interleukin-15. Exp Hematol (2008) 36(5):598-608. doi: 10.1016 / j.exphem.2008.01.001). Conversely, IL-15 is indispensable for the later development of NK cells. The expression of CD122 endows NK cells with the capacity to be responsive to IL-15; thus, these cells can become phenotypically and functionally mature and exhibit survival in response to IL-15 (16).[000101] IL-15 Receptor Expression Varies in Different Stages ofNK Cell Development. The expression of CD 122 on NK cells is not static but dynamically changes with NK cell maturation. It has been previously demonstrated that CD56brightNK cells express higher levels of CD122 as well as elevated CD 122 transcripts compared with CD56dimNK cells, and thus are intrinsically more responsive to IL-15 (Rautela J, Huntington ND. IL-15 signaling in NK cell cancer immunotherapy. Curr Opin Immunol (2017) 44: 1-6. doi: 10.1016 / j .coi.2016.10.004; Poli A, et al. CD56brightnatural killer (NK) cells: an important NK cell subset. Immunology (2009) 126(4):458- 65. doi: 10.111 l / j.l365-2567.2008.03027.x; Michel T. et al. Human CD56bright NK Cells: An Update. J Immunol (2016) 196(7):2923-31. doi: 10.4049 / jimmunol.1502570). This observation explains the decreased proliferation capacity in response to IL- 15 or dendritic cell (DC) stimulation during NK cell maturation and is consistent with the fact that cytokines, such as IL-2 and IL- 15, fail to reverse the proliferation defects of CD57+terminally matured NK cells (Lopez-Verges S.,et al. CD57 defines a functionally distinct population of mature NK cells in the human CD56dimCD16+NK-cell subset. Blood (2010) 116(19):3865-74. doi: 10.1182 / blood-2010-04- 282301). Consistent with the observation in human NK cells, CD122 expression is significantly decreased during maturation from mice CD1 lb+CD27+NK cells to CD1 lb CD27 NK cells and concomitant with decreased proliferation capacity (Yang C., et al. mTORCl and mT0RC2 differentially promote natural killer cell development. Elife (2018) 7:e35619. doi: 10.7554 / eLife.35619).[000102] Transcriptional Regulation of IL- 15 Receptor Expression at Different Stages During NK Cell Development. Although CD122 (encoded by I12rb) is critical for NK cell development by transducing IL-15 signaling, the coordinated regulation of CD122 expression by various transcription factors remains elusive. Studies have demonstrated that RUNX3 (one of the Runx family transcription factors), T-bet, and Eomesodermin (Eomes) directly bind to the promoter region of I12rb and induce CD122 expression (Ohno S-i, Sato T., et al. Runx proteins are involved in regulation of CD122, Ly49 family and IFN-gamma expression during NK cell differentiation. Int Immunol (2008) 20(1 ):71— 9. doi: 10.1093 / intimm / dxml20; Intlekofer AM., et al. Effector and memory CD8(+) T cell fate coupled by T-bet and eomesodermin. Nat Immunol (2005) 6(12): 1236-44. doi: 10.1038 / nil268). However, these transcription factors are not simultaneously functional, but rather function at different stages of NK cell development. In the NK cell development pathway, RUNX3 expression is initiated at the NKP stage. The inactivation of RUNX3 in HSCs partially disturbed the generation of CD122+NKP cells in vitro but not completely, indicating that other unknown transcription factors contribute to the expression of CD122 during NK cell commitment. In addition, the deletion of RUNX3 in immature NK cells in mice only slightly reduced CD122 expression on NK cells, and the absolute number of NK cells was not significantly affected. These results confirmed that RUNX3 is necessary for the acquisition of CD 122 during NK cell lineage commitment but is not essential for the maintenance of CD 122 at the later maturation stages of NK cell development (Ohno S-i, Sato T., et al. 2005).[000103] In contrast, T-bet and Eomes are weakly expressed at the NKP stage but highly expressed during NK cell maturation (Gordon SM., et al. The transcription factors T-bet and Eomes control key checkpoints of natural killer cell maturation. Immunity (2012) 36(l):55-67.doi: 10.1016 / j.immuni.2011 .11 .016). Consistently, mice harboring genomic deletions of T-bet and Eomes lack NK cells, but CD 122hlprecursors ofNK cells were observed. In addition, the deletion of Eomes in mice results in significantly decreased CD122 expression at different stages of NK cell maturation (Wagner JA, et al. Stage-Specific Requirement for Eomes in Mature NK Cell Homeostasis and Cytotoxicity. Cell Rep (2020) 31(9): 107720. doi:10.1016 / j.celrep.2020.107720). Moreover, Eomes+NK cells express more CD122 and proliferate better than Eomes NK cells, which are called Innate Lymphoid Cells (ILCs) 1 now (Colonna M. Innate Lymphoid Cells: Diversity, Plasticity, and Unique Functions in Immunity. Immunity (2018) 48(6): 1104-17. doi: 10.1016 / j.immuni.2018.05.013). However, CD122 expression is upregulated in T-bet-deficient NK cells, and this finding may be attributed to increased Eomes expression, which is repressed by T-bet (van Helden MJ, et al. Terminal NK cell maturation is controlled by concerted actions of T-bet and Zeb2 and is essential for melanoma rejection. J Exp Med (2015) 212(12):2015- 25. doi: 10.1084 / jem.20150809). These results indicate that Eomes but not T-bet plays a dominant role in the maintenance of CD 122 expression during NK cell maturation. Consistently, although T-bet expression is upregulated during the NK cell transition from the CDl lb+CD27+to CD I Ib CD27 stage, CD122 expression is progressively decreased, accompanied by a reduction in Eomes expression.[000104] Accordingly, embodiments of this disclosure are directed to CCRs with antigenspecific cytokine signaling to increase NK persistence and confer powerful long-term anti-AML efficacy while avoiding in vivo toxicity. By utilizing heterodimerized complexes in the same fashion as native cytokine signaling, the compositions embodied herein surmount current clinical limitations of NK cell therapies.[000105] In certain aspects, a chimeric cytokine receptor comprises (i) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor domains, an interleukin receptor transmembrane domain, and an interleukin receptor intracellular domain; (ii) a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor common gamma chain (yc) domains, a yc transmembrane domain, and a yc intracellular domain. In certain embodiments, the one or more extracellular interleukin receptor domains comprise one or more interleukin 15 receptor (IL-15R)extracellular domains or variants thereof. In certain embodiments, the one or more transmembrane interleukin receptor domains comprise one or more interleukin 15 receptor (IL-15R) transmembrane domains or variants thereof. In certain embodiments, the one or more intracellular interleukin receptor domains comprise one or more interleukin 15 receptor (IL-15R) intracellular domains or variants thereof. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 50% shorter than a wild type IL- 15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 75% shorter than a wild type IL- 15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 90% shorter than a wild type IL- 15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 95% shorter than a wild type IL- 15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 99% shorter than a wild type IL- 15R extracellular domain. In certain embodiments, the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is as long as a wild type IL-15R extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 50% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 75% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 90% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 95% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 99% shorter than a wild type yc extracellular domain. In certain embodiments, the one or more extracellular domains of the yc comprise an amino acid sequence length that is as long as a wild type yc extracellular domain.[000106] Compositions herein include IL-15Rp variants of the present disclosure comprising an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the wild-type IL-15RP amino acid sequence (SEQ ID NO: 9; UniProtKB / Swiss-Prot:P14784; EMBL Accession CR456506.1 ; GenBank: CAG30392.1):[000107] Compositions herein include common gamma chain (yc) (cytokine receptor common subunit gamma) variants of the present disclosure comprising an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the common gamma chain amino acid sequence (SEQ ID NO: 9 A; UniProtKB / Swiss-Prot:P31785:[000108] In certain embodiments, the chimeric yc cytokine receptor comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 2 (includes a signal sequence):[000109] In certain embodiments, the chimeric P cytokine receptor comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 4 (includes a signal sequence):[000110] In the above SEQ ID. NO 4, the following end portion is a protein tag:[000111] In certain embodiments, the chimeric 0 cytokine receptor comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, atleast 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 4 A. (includes a signal sequence):[000112] In certain embodiments, the chimeric yc cytokine receptor comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 5 (SEQ ID NO: 2 minus the signal sequence):[000113] In the above SEQ ID. NO 5, the following end portion is a protein tag:[000114] In certain embodiments, the chimeric yc cytokine receptor comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 5A:[000115] In certain embodiments, the chimeric 0 cytokine receptor comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 6 (SEQ ID NO: 4 minus the signal sequence):[000116] In the above SEQ ID. NO 6, the following end portion is a protein tag:[000117] In certain embodiments, the chimeric 0 cytokine receptor comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 6A:[0001181 Incertain embodiments, a signal sequence comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 7: MLKPSLPFTSLLFLQLPLLGVG.[000119] In certain embodiments, a signal sequence comprises an amino acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 8: MDWIWRILFLVGAATGAHS .[000120] In certain embodiments, the IL-15RP molecules comprises one or more modified amino acids, unnatural amino acids, substituted amino acids or combinations thereof Accordingly, the cytokine receptor comprising SEQ ID NOs: 2, 4, 5 or 6 further comprises one or more modified amino acids, unnatural amino acids, substituted amino acids or combinations thereof. Nonlimiting examples of non-natural amino acids include selenocysteine, pyrrolysine, homocysteine, an O-methyl-L-tyrosine, an L-3-(2-naphthyl)alanine, a 3-methyl-phenylalanine, an O-4-allyl-L- tyrosine, a 4-propyl-L-tyrosine, a tri-O-acetyl-GlcNAcP-serine, an L-Dopa, a fluorinated phenylalanine, an isopropyl-L-phenylalanine, a p-azido-L-phenylalanine, a p-acyl-L- phenylalanine, a p-benzoyl-L-phenylalanine, an L-phosphoserine, a phosphonoserine, a phosphonotyrosine, a p-iodo-phenylalanine, a p-bromophenylalanine, a p-amino-L-phenylalanine, an isopropyl-L-phenylalanine, an unnatural analogue of a tyrosine amino acid; an unnaturalanalogue of a glutamine amino acid; an unnatural analogue of a phenylalanine amino acid; an unnatural analogue of a serine amino acid; an unnatural analogue of a threonine amino acid; an alkyl, aryl, acyl, azido, cyano, halo, hydrazine, hydrazide, hydroxyl, alkenyl, alkynl, ether, thiol, sulfonyl, seleno, ester, thioacid, borate, boronate, phospho, phosphono, phosphine, heterocyclic, enone, imine, aldehyde, hydroxylamine, keto, or amino substituted amino acid, or any combination thereof; a fluorescent amino acid; an amino acid with a novel functional group; an amino acid that covalently or noncovalently interacts with another molecule; a metal binding amino acid; a metalcontaining amino acid; a radioactive amino acid; a photocaged and / or photoisomerizable amino acid; a biotin or biotin-analogue containing amino acid; a glycosylated or carbohydrate modified amino acid; a keto containing amino acid; amino acids comprising polyethylene glycol or polyether; a heavy atom substituted amino acid; a chemically cleavable or photocleavable amino acid; an amino acid with an elongated side chain; a sugar substituted amino acid, e.g., a sugar substituted serine or the like; a carbon-linked sugar-containing amino acid; a redox-active amino acid; an a-hydroxy containing acid; an amino thio acid containing amino acid; an a, a disubstituted amino acid; a P-amino acid; and a cyclic amino acid other than proline.[000121] Antigen Specific Binding Domain[000122] In certain embodiments, the antigen specific binding domain specifically binds to tumor or cancer antigens, virus antigens or combinations thereof. In certain embodiments, the antigen specific binding domain specifically binds cluster of differentiation antigen 123 (CD 123). In certain embodiments, the antigen specific binding domain comprises an antibody, an antibody binding fragment, an aptamer, or a peptide. In certain embodiments, the antigen specific binding domain comprises a single chain variable fragment (scFv).[000123] Wild type CD123 (SEQ ID NO: 10; Uniprot P26951-1):[000124] In certain embodiments, the antigen specific binding domain specifically binds cluster of differentiation B7H3 (CD276). In certain embodiments, the antigen specific binding domain comprises an antibody, an antibody binding fragment, an aptamer, or a peptide.[000125] Human CD276 (SEQ ID NO: 10A; Uniport Q5ZPR3-1):[000126] Numerous antigen-binding domains are known in the art, including those based on the antigen binding site of an antibody, antibody mimetics, nanobodies, and T-cell receptor fragments. For example, the antigen-binding domain may comprise: a single-chain variable fragment (scFv) derived from a monoclonal antibody; a natural ligand of the target antigen; a peptide with sufficient affinity for the target; a single domain binder such as a camelid; an artificial binder single as a Darpin; or a single-chain derived from a T-cell receptor. Accordingly, the antigen specific binding domain includes, without limitation, an antibody, a T cell receptor fragment, a soluble T cell receptor, nanobody, aptamer, syn / notch recognition domain / effector domain pair, receptors, fragments or combinations thereof. In certain embodiments, the antigen binding domain is a single chain variable fragment (scFv).[000127] In certain embodiments, the antigen binding domain is or comprises an antibody or antibody fragment, aptamers, proteins and the like. In certain embodiments, the antibodies are human antibodies, including any known to bind a targeting molecule. The term “antibody” herein is used in the broadest sense and includes polyclonal and monoclonal antibodies, including intact antibodies and functional (antigen-binding) antibody fragments, including fragment antigen binding (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rlgG) fragments, variable heavy chain (VH) regions capable of specifically binding the antigen, singlechain antibody fragments, including single chain variable fragments (scFv), and single domain antibodies (e.g., sdAb, sdFv, nanobody) fragments. The term encompasses genetically engineered and / or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific, e.g., bispecific, antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise stated, the term “antibody” should be understood to encompass functional antibody fragments thereof. The term also encompasses intact or full-length antibodies, including antibodies of any class or sub-class, including IgG and sub-classes thereof, IgM, IgE, IgA, and IgD.[000128] In some embodiments, the antigen-binding domain is a humanized antibody of fragments thereof. A humanized antibody optionally may include at least a portion of an antibody constant region derived from a human antibody. A “humanized form” of a non-human antibody, refers to a variant of the non-human antibody that has undergone humanization, typically to reduce immunogenicity to humans, while retaining the specificity and affinity of the parental non-human antibody. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., the antibody from which the CDR residues are derived), e.g., to restore or improve antibody specificity or affinity.[000129] In some embodiments, the heavy and light chains of an antibody can be full-length or can be an antigen-binding portion (a Fab, F(ab')2, Fv or a single chain Fv fragment (scFv)). In other embodiments, the antibody heavy chain constant region is chosen from, e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE, particularly chosen from, e.g., IgGl, IgG2, IgG3, and IgG4, more particularly, IgGl (e.g., human IgGl). In another embodiment, the antibody light chain constant region is chosen from, e g., kappa or lambda, particularly kappa.[000130] Among the provided antibodies are antibody fragments. Examples of antibody fragments include but are not limited to Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; variable heavy chain (Vn) regions, single-chain antibody molecules such as scFvs and single-domain VH single antibodies; and multispecific antibodies formed from antibody fragments. In particular embodiments, the antibodies are single-chain antibody fragments comprising a variable heavy chain region and / or a variable light chain region, such as scFvs. Single-domainantibodies are antibody fragments comprising all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody.[000131] Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody as well as production by recombinant host cells. In some embodiments, the antibodies are recombinantly produced fragments, such as fragments comprising arrangements that do not occur naturally, such as those with two or more antibody regions or chains joined by synthetic linkers, e.g., peptide linkers, and / or that are may not be produced by enzyme digestion of a naturally-occurring intact antibody. In some aspects, the antibody fragments are scFvs.[000132] Cancer Antigens: As used herein, a cancer antigen is an antigenic substance present in cancer cells. In principle, any protein produced in a cancer cell that has an abnormal structure due to mutation can act as a cancer antigen. In principle, cancer antigens can be products of mutated oncogenes and tumor suppressor genes, products of other mutated genes, overexpressed or aberrantly expressed cellular proteins, cancer antigens produced by oncogenic viruses, oncofetal antigens, altered cell surface glycolipids and glycoproteins, or cell type-specific differentiation antigens. Examples of cancer antigens include the abnormal products of ras and p53 genes. Other examples include tissue differentiation antigens, mutant protein antigens, oncogenic viral antigens, cancer-testis antigens and vascular or stromal specific antigens. Tissue differentiation antigens are those that are specific to a certain type of tissue. Mutant protein antigens are likely to be much more specific to cancer cells because normal cells should not contain these proteins. Normal cells will display the normal protein antigen on their MHC molecules, whereas cancer cells will display the mutant version. Some viral proteins are implicated in forming cancer, and some viral antigens are also cancer antigens. Cancer-testis antigens are antigens expressed primarily in the germ cells of the testes, but also in fetal ovaries and the trophoblast. Some cancer cells aberrantly express these proteins and therefore present these antigens, allowing attack by T-cells specific to these antigens. Exemplary antigens of this type are CTAG1 B and MAGEA1 as well as Rindopepimut, a 14-mer intradermal injectable peptide vaccine targeted against epidermal growth factor receptor (EGFR) vIII variant. Rindopepimut is particularly suitable for treating glioblastoma when used incombination with an inhibitor of the CD95 / CD95L signaling system as described herein. Also, proteins that are normally produced in very low quantities, but whose production is dramatically increased in cancer cells, may trigger an immune response. An example of such a protein is the enzyme tyrosinase, which is required for melanin production. Normally tyrosinase is produced in minute quantities but its levels are very much elevated in melanoma cells. Oncofetal antigens are another important class of cancer antigens. Examples are alpha fetoprotein (AFP) and carcinoembryonic antigen (CEA). These proteins are normally produced in the early stages of embryonic development and disappear by the time the immune system is fully developed. Thus, self-tolerance does not develop against these antigens. Abnormal proteins are also produced by cells infected with oncoviruses, e.g. EBV and HPV. Cells infected by these viruses contain latent viral DNA which is transcribed, and the resulting protein produces an immune response. A cancer vaccine may include a peptide cancer vaccine, which in some embodiments is a personalized peptide vaccine. In some embodiments, the peptide cancer vaccine is a multivalent long peptide vaccine, a multi-peptide vaccine, a peptide cocktail vaccine, a hybrid peptide vaccine, or a peptide- pulsed dendritic cell vaccine[000133] Accordingly, in certain embodiments, the antigen-specific binding domain binds to an antigen on a tumor cell. Tumor-specific binding domains may be derived from antibodies approved for treatment of patients with cancer include rituximab, ofatumumab, and obinutuzumab (anti-CD20 Abs); trastuzumab and pertuzumab (anti-HER2 Abs); cetuximab and panitumumab (anti-EGFR Abs); and alemtuzumab (anti-CD52 Ab). Similarly, binding domains from approved antibody-effector molecule conjugates specific to CD20 (90Y-labeled ibritumomab tiuxetan, 1311- labeled tositumomab), HER2 (ado-trastuzumab emtansine), CD30 (brentuximab vedotin) and CD33 (gemtuzumab ozogamicin) (Sliwkowski MX, Mellman I. 2013 Science 341 : 1192) could be used.[000134] Additionally, binding domains of the disclosure may include various other tumorspecific antibody domains known in the art. The antibodies and their respective targets for treatment of cancer include but are not limited to nivolumab (anti-PD-1 Ab), TA99 (anti-gp75), 3F8 (anti-GD2), 8H9 (anti-B7-H3), abagovomab (anti-CA-125 (imitation)), adecatumumab (anti- EpCAM), afutuzumab (anti-CD20), alacizumab pegol (anti-VEGFR2), altumomab pentetate (anti-CEA), amatuximab (anti-mesothelin), AME-133 (anti-CD20), anatumomab mafenatox (anti- TAG-72), apolizumab (anti-HLA-DR), arcitumomab (anti-CEA), bavituximab (antiphosphatidylserine), bectumomab (anti-CD22), belimumab (anti-BAFF), besilesomab (anti-CEA- related antigen), bevacizumab (anti-VEGF-A), bivatuzumab mertansine (anti-CD44 v6), blinatumomab (anti-CD19), BMS-663513 (anti-CD137), brentuximab vedotin (anti-CD30 (TNFRSF8)), cantuzumab mertansine (anti -mucin CanAg), cantuzumab ravtansine (anti-MUCl), capromab pendetide (anti-prostatic carcinoma cells), carlumab (anti-MCP-1), catumaxomab (anti- EpCAM, CD3), cBR96-doxorubicin immunoconjugate (anti-Lewis-Y antigen), CC49 (anti-TAG- 72), cedelizumab (anti-CD4), Ch.14.18 (anti-GD2), ch-TNT (anti-DNA associated antigens), citatuzumab bogatox (anti-EpCAM), cixutumumab (anti -IGF- 1 receptor), clivatuzumab tetraxetan (anti-MUCl), conatumumab (anti-TRAIL-R2), CP-870893 (anti-CD40), dacetuzumab (anti- CD40), daclizumab (anti-CD25), dalotuzumab (anti-insulin-like growth factor I receptor), daratumumab (anti-CD38 (cyclic ADP ribose hydrolase)), demcizumab (anti-DLL4), detumomab (anti-B-lymphoma cell), drozitumab (anti-DR5), duligotumab (anti-HER3), dusigitumab (anti- ILGF2), ecromeximab (anti-GD3 ganglioside), edrecolomab (anti-EpCAM), elotuzumab (anti- SLAMF7), elsilimomab (anti-IL-6), enavatuzumab (anti-TWEAK receptor), enoticumab (anti- DLL4), ensituximab (anti-5 AC), epitumomab cituxetan (anti-epi sialin), epratuzumab (anti-CD22), ertumaxomab (anti-HER2 / neu, CD3), etaracizumab (anti-integrin av03), faralimomab (anti- Interferon receptor), farletuzumab (anti-folate receptor 1), FBTA05 (anti-CD20), ficlatuzumab (anti-HGF), figitumumab (anti-IGF-1 receptor), flanvotumab (anti -TYRP1 (glycoprotein 75)), fresolimumab (anti-TGF P), futuximab (anti-EGFR), galiximab (anti-CD80), ganitumab (anti- IGF-I), gemtuzumab ozogamicin (anti-CD33), girentuximab (anti-carbonic anhydrase 9 (CA-IX)), glembatumumab vedotin (anti-GPNMB), guselkumab (anti-IL13), ibalizumab (anti-CD4), ibritumomab tiuxetan (anti-CD20), icrucumab (anti-VEGFR-1), igovomab (anti-CA-125), IMAB362 (anti-CLDN18.2), IMC-CS4 (anti-CSFIR), IMC-TR1 (TGFpRII), imgatuzumab (anti- EGFR), inclacumab (anti-selectin P), indatuximab ravtansine (anti-SDCl), inotuzumab ozogamicin (anti-CD22), intetumumab (anti-CD51), ipilimumab (anti-CD152), iratumumab (anti- CD30 (TNFRSF8)), KM3065 (anti-CD20), KW-0761 (anti-CD194), LY2875358 (anti-MET) labetuzumab (anti-CEA), lambrolizumab (anti-PDCDl), lexatumumab (anti -TRAIL -R2),lintuzumab (anti-CD33), lirilumab (anti-KTR2D), lorvotuzumab mertansine (anti-CD56), lucatumumab (anti-CD40), lumiliximab (anti-CD23 (IgE receptor)), mapatumumab (anti-TRAIL- Rl), margetuximab (anti-ch4D5), matuzumab (anti-EGFR), mavrilimumab (anti-GMCSF receptor a-chain), milatuzumab (anti-CD74), minretumomab (anti-TAG-72), mitumomab (anti-GD3 ganglioside), mogamulizumab (anti-CCR4), moxetumomab pasudotox (anti-CD22), nacolomab tafenatox (anti-C242 antigen), naptumomab estafenatox (anti-5T4), narnatumab (anti-RON), necitumumab (anti-EGFR), nesvacumab (anti-angiopoietin 2), nimotuzumab (anti-EGFR), nivolumab (anti-IgG4), nofetumomab merpentan, ocrelizumab (anti-CD20), ocaratuzumab (anti- CD20), olaratumab (anti-PDGF-R a), onartuzumab (anti-c-MET), ontuxizumab (anti-TEMl), oportuzumab monatox (anti-EpCAM), oregovomab (anti-CA-125), otlertuzumab (anti-CD37), pankomab (anti-tumor specific glycosylation of MUC1), parsatuzumab (anti-EGFL7), pascolizumab (anti-IL-4), patritumab (anti-HER3), pemtumomab (anti-MUCl), pertuzumab (anti- HER2 / neu), pidilizumab (anti-PD-1), pinatuzumab vedotin (anti-CD22), pintumomab (antiadenocarcinoma antigen), polatuzumab vedotin (anti-CD79B), pritumumab (anti-vimentin), PROD 1921 (anti-CD20), quilizumab (anti-IGHE), racotumomab (anti-N-glycolylneuraminic acid), radretumab (anti-fibronectin extra domain-B), ramucirumab (anti-VEGFR2), rilotumumab (anti-HGF), robatumumab (anti-IGF-1 receptor), roledumab (anti-RHD), rovelizumab (anti-CDl l & CD 18), samalizumab (anti-CD200), satumomab pendetide (anti-TAG-72), seribantumab (anti- ERBB3), SGN-CD19A (anti-CD19), SGN-CD33A (anti-CD33), sibrotuzumab (anti-FAP), siltuximab (anti-IL-6), solitomab (anti-EpCAM), sontuzumab (anti-epi sialin), tabalumab (anti- BAFF), tacatuzumab tetraxetan (anti -alpha-fetoprotein), taplitumomab paptox (anti-CD19), telimomab aritox, tenatumomab (anti-tenascin C), teneliximab (anti-CD40), teprotumumab (anti- CD221), TGN1412 (anti-CD28), ticilimumab (anti-CTLA-4), tigatuzumab (anti-TRAIL-R2), TNX-650 (anti-IL-13), tositumomab (anti-CS20), tovetumab (anti-CD140a), TRBS07 (anti-GD2), tregalizumab (anti-CD4), tremelimumab (anti-CTLA-4), TRU-016 (anti-CD37), tucotuzumab celmoleukin (anti-EpCAM), ublituximab (anti-CD20), urelumab (anti-4-lBB), vantictumab (anti- Frizzled receptor), vapaliximab (anti-AOC3 (VAP-1)), vatelizumab (anti-ITGA2), veltuzumab (anti-CD20), vesencumab (anti-NRPl), visilizumab (anti-CD3), volociximab (anti-integrin a501), vorsetuzumab mafodotin (anti-CD70), votumumab (anti-tumor antigen CTAA16.88),zalutumumab (anti-EGFR), zanolimumab (anti-CD4), zatuximab (anti-HERl), ziralimumab (anti- CD147 (basigin)), RG7636 (anti-ETBR), RG7458 (anti-MUC16), RG7599 (anti-NaPi2b), MPDL3280A (anti-PD-Ll), RG7450 (anti-STEAPl), and GDC-0199 (anti-Bcl-2).[000135] The antibodies and their respective targets for treatment of cancer include 8H9 (anti-B7-H3).[000136] Other antibody domains or tumor target binding proteins useful in the disclosure include, but are not limited to, those that bind the following antigens (note, the cancer indications indicated represent non-limiting examples): aminopeptidase N (CD13), annexin Al, B7-H3 (CD276, various cancers), CA125 (ovarian cancers), CA15-3 (carcinomas), CA19-9 (carcinomas), L6 (carcinomas), Lewis Y (carcinomas), Lewis X (carcinomas), alpha fetoprotein (carcinomas), CA242 (colorectal cancers), placental alkaline phosphatase (carcinomas), prostate specific antigen (prostate), prostatic acid phosphatase (prostate), epidermal growth factor (carcinomas), CD2 (Hodgkin's disease, NHL lymphoma, multiple myeloma), CD3 epsilon (T cell lymphoma, lung, breast, gastric, ovarian cancers, autoimmune diseases, malignant ascites), CD 19 (B cell malignancies), CD20 (non-Hodgkin's lymphoma, B-cell neoplasmas, autoimmune diseases), CD21 (B-cell lymphoma), CD22 (leukemia, lymphoma, multiple myeloma, SLE), CD30 (Hodgkin's lymphoma), CD33 (leukemia, autoimmune diseases), CD38 (multiple myeloma), CD40 (lymphoma, multiple myeloma, leukemia (CLL)), CD51 (metastatic melanoma, sarcoma), CD52 (leukemia), CD56 (small cell lung cancers, ovarian cancer, Merkel cell carcinoma, and the liquid tumor, multiple myeloma), CD66e (carcinomas), CD70 (metastatic renal cell carcinoma and non-Hodgkin lymphoma), CD74 (multiple myeloma), CD80 (lymphoma), CD98 (carcinomas), CD 123 (leukemia), mucin (carcinomas), CD221 (solid tumors), CD227 (breast, ovarian cancers), CD262 (NSCLC and other cancers), CD309 (ovarian cancers), CD326 (solid tumors), CEACAM3 (colorectal, gastric cancers), CEACAM5 (CEA, CD66e) (breast, colorectal and lung cancers), DLL4 (A-like-4), EGFR (various cancers), CTLA4 (melanoma), CXCR4 (CD 184, hemeoncology, solid tumors), Endoglin (CD 105, solid tumors), EPCAM (epithelial cell adhesion molecule, bladder, head, neck, colon, NHL prostate, and ovarian cancers), ERBB2 (lung, breast, prostate cancers), FCGR1 (autoimmune diseases), FOLR (folate receptor, ovarian cancers), FGFR (carcinomas), GD2 ganglioside (carcinomas), G-28 (a cell surface antigen glycolipid, melanoma),GD3 idiotype (carcinomas), heat shock proteins (carcinomas), HER1 (lung, stomach cancers), HER2 (breast, lung and ovarian cancers), HLA-DR10 (NHL), HLA-DRB (NHL, B cell leukemia), human chorionic gonadotropin (carcinomas), IGF1R (solid tumors, blood cancers), IL-2 receptor (T-cell leukemia and lymphomas), IL-6R (multiple myeloma, RA, Castleman's disease, IL6 dependent tumors), integrins (av03, a501, a604, al ip3, a5p5, avP5, for various cancers), MAGE- 1 (carcinomas), MAGE-2 (carcinomas), MAGE-3 (carcinomas), MAGE 4 (carcinomas), antitransferrin receptor (carcinomas), p97 (melanoma), MS4A1 (membrane-spanning 4-domains subfamily A member 1, Non-Hodgkin's B cell lymphoma, leukemia), MUC1 (breast, ovarian, cervix, bronchus and gastrointestinal cancer), MUC16 (CAI 25) (ovarian cancers), CEA (colorectal cancer), gplOO (melanoma), MARTI (melanoma), MPG (melanoma), MS4A1 (membrane-spanning 4-domains subfamily A, small cell lung cancers, NHL), nucleolin, Neu oncogene product (carcinomas), P21 (carcinomas), nectin-4 (carcinomas), paratope of anti-(N- glycolylneuraminic acid, breast, melanoma cancers), PLAP-like testicular alkaline phosphatase (ovarian, testicular cancers), PSMA (prostate tumors), PSA (prostate), ROB04, TAG 72 (tumor associated glycoprotein 72, AML, gastric, colorectal, ovarian cancers), T cell transmembrane protein (cancers), Tie (CD202b), tissue factor, TNFRSF10B (tumor necrosis factor receptor superfamily member 10B, carcinomas), TNFRSF13B (tumor necrosis factor receptor superfamily member 13B, multiple myeloma, NHL, other cancers, RA and SLE), TPBG (trophoblast glycoprotein, renal cell carcinoma), TRAIL-R1 (tumor necrosis apoptosis inducing ligand receptor 1, lymphoma, NHL, colorectal, lung cancers), VCAM-1 (CD106, Melanoma), VEGF, VEGF-A, VEGF-2 (CD309) (various cancers). Some other tumor associated antigen targets have been reviewed (Gerber, et al, mAbs 2009 1 :247-253; Novellino et al., Cancer Immunol Immunother. 2005 54: 187-207, Franke, et al., Cancer Biother Radiopharm. 2000, 15:459-76, Guo, et al., Adv Cancer Res. 2013; 119: 421-475, Parmiani et al., J Immunol. 2007 178: 1975-9). Examples of these antigens include Cluster of Differentiations (CD4, CD5, CD6, CD7, CD8, CD9, CD 10, CDl la, CDl lb, CDllc, , CD12w, CD14, CD15, CD16, CDwl7, CD18, CD21, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD31, CD32, CD34, CD35, CD36, CD37, CD41, CD42, CD43, CD44, CD45, CD46, CD47, CD48, CD49b, CD49c, CD53, CD54, CD55, CD58, CD59, CD61, CD62E, CD62L, CD62P, CD63, CD68, CD69, CD71, CD72, CD79, CD81, CD82, CD83,CD86, CD87, CD88, CD89, CD90, CD91, CD95, CD96, CD 100, CD 103, CD 105, CD 106, CD109, CD117, CD120, CD123, CD127, CD133, CD134, CD135, CD138, CD141, CD142, CD143, CD144, CD147, CD151, CD152, CD154, CD156, CD158, CD163, CD166, .CD168, CD184, CDwl86, CD195, CD202 (a, b), CD209, CD235a, CD271, CD303, CD304), annexin Al, nucleolin, endoglin (CD 105), ROB04, amino-peptidase N, -like-4 (DLL4), VEGFR-2 (CD309), CXCR4 (CD184), Tie2, B7-H3, WT1, MUC1, LMP2, HPV E6 E7, EGFRvIII, HER-2 / neu, idiotype, MAGE A3, p53 nonmutant, NY-ESO-1, GD2, CEA, MelanA / MARTl, Ras mutant, gplOO, p53 mutant, proteinase3 (PR1), bcr-abl, tyrosinase, survivin, hTERT, sarcoma translocation breakpoints, EphA2, PAP, ML-IAP, AFP, EpCAM, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, ALK, androgen receptor, cyclin B 1, polysialic acid, MYCN, RhoC, TRP-2, GD3, fucosyl GM1 , mesothelin, PSCA, MAGE Al, sLe(a), CYPIB I, PLAC1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, carbonic anhydrase IX, PAX5, OY-TES1, sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, B7H3, legumain, Tie 2, Page4, VEGFR2, MAD- CT-1, FAP, PDGFR-P, MAD-CT-2, Notch 1, ICAM1 and Fos-related antigen 1.[000137] In certain embodiments, the antigen binding domain specifically binds to a virus antigen. Non- limiting examples of such antigens include antigens of the flavivirus, West Nile Virus (WNV) including structural proteins, e.g., C, M, and E, and non-structural proteins, e.g., NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5; human immunodeficiency virus (HIV) antigens gp41, gpl20, gpl60, Nef, Gag, and Rev, Tat, Vif, Vpu, Vpr, or vpx; influenza virus hemagglutinin; human respiratory syncytial virus G glycoprotein; core protein, matrix protein or other protein of Dengue virus; measles virus hemagglutinin; herpes simplex virus type 2 glycoprotein gB; poliovirus I VP1 (Emini etal., 1983, Nature 304:699); an envelope glycoprotein of HIV I; hepatitis B surface antigen; diptheria toxin; streptococcus 24M epitope; gonococcal pilin; pseudorabies virus g50 (gpD); pseudorabies virus II (gpB); pseudorabies virus gill (gpC); pseudorabies virus glycoprotein H; pseudorabies virus glycoprotein E; transmissible gastroenteritis glycoprotein 195; transmissible gastroenteritis matrix protein; swine rotavirus glycoprotein 38; swine parvovirus capsid protein; Serpulina hydodysenteriae protective antigen; bovine viral diarrhea glycoprotein 55; Newcastle disease virus hemagglutinin-neuraminidase; swine flu hemagglutinin; swine flu neuraminidase; antigens of foot and mouth disease virus; antigens of hog cholera virus; antigensof swine influenza virus; antigens of African swine fever virus; Mycoplasma hyopneumoniae; antigens of infectious bovine rhinotracheitis virus {e.g. , infectious bovine rhinotracheitis virus glycoprotein E or glycoprotein G); antigens of infectious laryngotracheitis virus {e.g. , infectious laryngotracheitis virus glycoprotein G or glycoprotein I); a glycoprotein of La Crosse virus; antigens of neonatal calf diarrhea virus; Venezuelan equine encephalomyelitis virus; punta toro virus; murine leukemia virus; mouse mammary tumor virus; hepatitis B virus core protein and / or hepatitis B virus surface antigen or a fragment or derivative thereof (see, e.g., Ganem and Varmus, 1987, Ann. Rev. Biochem. 56:651-693); antigen of equine influenza virus or equine herpesvirus (e.g., equine influenza virus type AJ Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase; equine influenza virus type A / Kentucky 81 neuraminidase; equine herpes virus type 1 glycoprotein B; equine herpes virus type 1 glycoprotein D); antigen of bovine respiratory syncytial virus or bovine parainfluenza virus {e.g., bovine respiratory syncytial virus attachment protein (BRSV G); bovine respiratory syncytial virus fusion protein (BRSV F); bovine respiratory syncytial virus nucleocapsid protein (BRSV N); bovine parainfluenza virus type 3 fusion protein; the bovine parainfluenza virus type 3 hemagglutinin neuraminidase); bovine viral diarrhea virus glycoprotein 48 or glycoprotein 53.[000138] NUCLEIC ACIDS AND VECTORS[000139] Provided herein are nucleic acids that encode IL-15RP and yc chains wherein the IL-15RP and yc chains can form a complex and bind to a specific antigen.[000140] In certain embodiments, the nucleic acids comprises a nucleic acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 1 : atgctgaagcccagcctgccttttaccagcctgctgttcctgcagctgcctctgcttggcgtgggccagattcagctggtgcagtctggcccc gagctgaagaaacccggcgagacagtgaagatcagctgcaaggccagcggctacatcttcaccaactacggcatgaactgggtcaagca ggcccctggcaagagcttcaagtggatgggctggatcaacacctacaccggcgagagcacctacagcgccgacttcaagggcagattcg[000141] In certain embodiments, the nucleic acids comprises a nucleic acid sequence that has at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 3 :[000142] In certain embodiments, the nucleic acids that encode IL-15RP and / or yc chain are optimized, e.g., by codon / RNA optimization, replacement with heterologous signal sequences, and elimination of mRNA instability elements. Methods to generate optimized nucleic acids encoding IL-15R0 and / or yc chain for expression by introducing codon changes and / or eliminating inhibitory regions in the mRNA can be carried out by adapting the optimization methods described in, e.g., U.S. Patent Nos. 5,965,726; 6,174,666; 6,291,664; 6,414,132; and 6,794,498. For example, potential splice sites and instability elements (e.g., A / T or A / U rich elements) within the RNA of IL-15RP and / or yc chain can be mutated without altering the amino acids encoded by the nucleic acid sequences to increase stability of the RNA for expression. The alterations utilize the degeneracy of the genetic code, e.g., using an alternative codon for an identical amino acid. Insome embodiments, it may be desirable to alter one or more codons to encode a conservative mutation, e.g., a similar amino acid with similar chemical structure and properties and / or function as the original amino acid. Such methods can increase expression of IL-15RP and / or yc chain proteins by at least 1 fold, 2 fold, 3 fold, 4 fold, 5 fold, 10 fold, 20 fold, 30 fold, 40 fold, 50 fold, 60 fold, 70 fold, 80 fold, 90 fold, or 100 fold or more relative to the expression of IL-15RP and / or yc chain proteins encoded by native nucleic acid sequences.[000143J Further, the native signal peptide sequence of IL-15RP and / or yc chain can be replaced with a heterologous signal peptide, e.g., a signal peptide of human GM-CSF, tissue plasminogen activator (tPA), preprolactin, growth hormone or an immunoglobulin protein (e.g., IgE).[000144] Vectors: In certain embodiments, a vector or genetic construct comprises adenovirus, adeno-associated virus (AAV), herpes simplex virus, gammaretrovirus lentivirus, retrovirus, alphavirus, flavivirus, rhabdovirus, measles virus, Newcastle disease virus, poxvirus, vaccinia virus, modified Ankara virus, or vesicular stomatitis virus.[000145] The vectors can be monocistronic or multicistronic. A multi ci stronic nucleic acid construct may encode 2, 3, 4, 5, 6, 7, 8, 9, 10 or more, or in the range of 2-5, 5-10 or 10-20 genes / nucleotide sequences. For example, a bicistronic nucleic acid construct may comprise in the following order a promoter, a first gene (e.g., IL-15RP), and a second gene and (e.g., the yc chain). In such a nucleic acid construct, the transcription of both genes is driven by the promoter, whereas the translation of the mRNA from the first gene is by a cap-dependent scanning mechanism and the translation of the mRNA from the second gene is by a cap-independent mechanism, e.g., by an IRES. In certain embodiments, the vector is a bicistronic vector.[000146] Accordingly, in certain embodiments, IL-15RP and the yc chain are encoded by one nucleic acid construct (e.g., bicistronic construct). In some embodiments, the IL-15RP and the yc chain are encoded by one nucleic acid construct comprising a single open reading frame (ORF) of IL-15RP and yc chain. In other embodiments, IL-15RP and the yc chain are encoded by two nucleic acid constructs, wherein a first nucleic acid construct encodes IL-15RP, a second nucleic acid construct encodes the yc chain. The IL-15RP encoded by the first nucleic acid construct comprisesa nucleic acid encoding a heterologous molecule, such as an antigen or an antibody of interest, e g. an antigen specific binding domain which specifically binds to CD 123. Alternatively, or in addition, the yc chain encoded by the second nucleic acid construct comprises a nucleic acid encoding an antigen specific binding domain which specifically binds to the same antigen as the IL-15R0, e.g., CD 123.[000147] Vectors can include, for example, origins of replication, scaffold attachment regions (SARs), and / or markers. A marker gene can confer a selectable phenotype on a host cell. For example, a marker can confer biocide resistance, such as resistance to an antibiotic (e.g., kanamycin, G418, bleomycin, or hygromycin). An expression vector can include a tag sequence designed to facilitate manipulation or detection (e.g., purification or localization) of the expressed polypeptide. Tag sequences, such as green fluorescent protein (GFP), glutathione S-transferase (GST), polyhistidine, c-myc, hemagglutinin, or FLAG™ tag (Kodak, New Haven, Conn.) sequences typically are expressed as a fusion with the encoded polypeptide. Such tags can be inserted anywhere within the polypeptide, including at either the carboxyl or amino terminus. For example, a tag sequence comprises SEQ ID NO: 11 :[000148] In certain embodiments, the IL-15R comprises (SEQ ID NO: 12):LVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKF EGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNI EDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAA GITLGMDELYK (Tag sequence in bold font)[000149] Additional expression vectors also can include, for example, segments of chromosomal, non-chromosomal and synthetic DNA sequences. Suitable vectors include derivatives of SV40 and known bacterial plasmids, e.g., E. coli plasmids col El, pCRl, pBR322, pMal-C2, pET, pGEX, pMB9 and their derivatives, plasmids such as RP4; phage DNAs, e.g., the numerous derivatives of phage 1, e.g., NM989, and other phage DNA, e.g., M13 and filamentous single stranded phage DNA; yeast plasmids such as the 2p plasmid or derivatives thereof, vectors useful in eukaryotic cells, such as vectors useful in insect or mammalian cells; vectors derived from combinations of plasmids and phage DNAs, such as plasmids that have been modified to employ phage DNA or other expression control sequences.[000150] The vector can also include a regulatory region. The term “regulatory region” refers to nucleotide sequences that influence transcription or translation initiation and rate, and stability and / or mobility of a transcription or translation product. Regulatory regions include, without limitation, promoter sequences, enhancer sequences, response elements, protein recognition sites, inducible elements, protein binding sequences, 5' and 3’ untranslated regions (UTRs), transcriptional start sites, termination sequences, polyadenylation sequences, nuclear localization signals, and introns.[000151] The term “operably linked” refers to positioning of a regulatory region and a sequence to be transcribed in a nucleic acid so as to influence transcription or translation of such a sequence. For example, to bring a coding sequence under the control of a promoter, the translation initiation site of the translational reading frame of the polypeptide is typically positioned between one and about fifty nucleotides downstream of the promoter. A promoter can, however, be positioned as much as about 5,000 nucleotides upstream of the translation initiation site or about 2,000 nucleotides upstream of the transcription start site. A promoter typically comprises at least a core (basal) promoter A promoter also may include at least one control element, such as an enhancer sequence, an upstream element or an upstream activation region (UAR), The choice of promoters to be included depends upon several factors, including, but notlimited to, efficiency, selectability, inducibility, desired expression level, and cell- or tissue- preferential expression. It is a routine mater for one of skill in the art to modulate the expression of a coding sequence by appropriately selecting and positioning promoters and other regulatory regions relative to the coding sequence.[000152] Vectors include, for example, viral vectors (such as adenoviruses (Ad), AAV, lenti virus, vesicular stomatitis virus (VSV), and retroviruses). Liposomes and other lipid- containing complexes, and other macromolecular complexes capable of mediating delivery of a polynucleotide to a host cell. Vectors can also comprise other components or functionalities that further modulate gene delivery and / or gene expression, or that otherwise provide beneficial properties to the targeted cells. Other components include, for example, components that influence binding or targeting to cells (including components that mediate cell-type or tissue-specific binding); components that influence uptake of the vector nucleic acid by the cell; components that influence localization of the polynucleotide within the cell after uptake (such as agents mediating nuclear localization); and components that influence expression of the polynucleotide. Such components also might include markers, such as detectable and / or selectable markers that can be used to detect or select for cells that have taken up and are expressing the nucleic acid delivered by the vector Such components can be provided as a natural feature of the vector (such as the use of certain viral vectors which have components or functionalities mediating binding and uptake), or vectors can be modified to provide such functionalities. Other vectors include those described by Chen etal:. Bio Techniques, 34: 167-171 (2003). A large variety of such vectors are known in the art and are generally available. A “recombinant viral vector” refers to a viral vector comprising one or more heterologous gene products or sequences. Since many viral vectors exhibit sizeconstraints associated with packaging, the heterologous gene products or sequences are typically introduced by replacing one or more portions of the viral genome Such viruses may become replication-defective, requiring the deleted function(s) to be provided in trans during viral replication and encapsidation (by using, e.g., a helper virus or a packaging cell line carrying gene products necessary for replication and / or encapsidation). Modified viral vectors in which a polynucleotide io be delivered is carried on the outside of the viral particle have also been described (see, e.g , Curiel, D T , ei cd. PHAS ES: 8850-8854, 1991).[000153] Additional vectors include viral vectors, fusion proteins and chemical conjugates Retroviral vectors include Moloney murine leukemia viruses and HIV-based viruses. One HIV based viral vector comprises at least two vectors wherein the gag and pol genes are from an HIV genome and the env gene is from another virus. DNA viral vectors include pox vectors such as orthopox or avipox vectors, herpesvirus vectors such as a herpes simplex I virus (HSV) vector [Geller, A. I. et al., J. Nettrochem, 64: 487 ( 1995); Lirn, F., et al., in DNA Cloning: Mammalian Systems, D. Glover, Ed. (Oxford Univ. Press, Oxford England) ( 1995); Geller, A. I. el al., Proc Natl. Acad. Sci.: U.S.A.;90 7603 (1993 ); Geller, A. I., et al., Proc Nail. Acad. Sc? USA: 87: 1 149 (1990)], Adenovirus Vectors [LeGal LaSalle etal., Science, 259:988 (1993); Davidson, el al., Nat. Genet. 3: 219 (1993); Yang, et al., J. Virol. 69. 2004 ( 1995)] and Adeno-associated Virus Vectors [Kaplitt, M. G., el al., Nat. Genet. 8: 148 (1994)].[000154] When taken up by a cell, the genetic construct which includes the nucleotide sequence encoding the desired protein operably linked to the regulatory elements may remain present in the cell as a functioning extrachromosomal molecule or it may integrate into the cell's chromosomal DNA. DNA may be introduced into cells where it remains as separate genetic material in the form of a plasmid. Alternatively, linear DNA which can integrate into the chromosome may be introduced into the cell. When introducing DNA into the cell, reagents which promote DNA integration into chromosomes may be added. DNA sequences which are useful to promote integration may also be included in the DNA molecule. Alternatively, RNA may be administered to the cell. It is also contemplated to provide the genetic construct as a linear minichromosome including a centromere, telomeres and an origin of replication.[000155] The regulatory elements necessary for gene expression of a DNA molecule include: a promoter, an initiation codon, a stop codon, and a polyadenylation signal. In addition, enhancers are often required for gene expression. It is necessary that these elements be operable linked to the sequence that encodes the desired proteins and that the regulatory elements are operably in the individual to whom they are administered.[000156] Initiation codons and stop codon are generally considered to be part of a nucleotide sequence that encodes the desired protein. However, it is necessary that these elements arefunctional in the individual to whom the gene construct is administered. The initiation and termination codons must be in frame with the coding sequence.[000157] Promoters and polyadenylation signals used must be functional within the cells of the individual. Examples of promoters useful to practice the present disclosure, especially in the production of a genetic vaccine for humans, include but are not limited to promoters from Simian Virus 40 (SV40, Mouse Mammary Tumor Virus (MMTV) promoter, Human Immunodeficiency Virus (HIV) such as the HIV Long Terminal Repeat (LTR) promoter, Moloney virus, ALV, Cytomegalovirus (CMV) such as the CMV immediate early promoter, Epstein Barr Virus (EBV), Rous Sarcoma Virus (RSV) as well as promoters from human genes such as human Actin, human Myosin, human Hemoglobin, human muscle creatine and human metalothionein. Examples of polyadenylation signals useful to practice the present disclosure, especially in the production of a genetic vaccine for humans, include but are not limited to SV40 polyadenylation signals and LTR polyadenylation signals.[000158] In addition to the regulatory elements required for DNA expression, other elements may also be included in the DNA molecule. Such additional elements include enhancers. The enhancer may be selected from the group including but not limited to: human Actin, human Myosin, human Hemoglobin, human muscle creatine and viral enhancers such as those from CMV, RSV and EBV. Genetic constructs can be provided with mammalian origin of replication in order to maintain the construct extrachromosomally and produce multiple copies of the construct in the cell. Plasmids pCEP4 and pREP4 from Invitrogen (San Diego, Calif.) contain the Epstein Barr virus origin of replication and nuclear antigen EBNA-1 coding region which produces high copy episomal replication without integration.[000159] In certain embodiments, a genetic construct may include a suicide gene. An example of a suicide gene is inducible human caspase-9 transgene (iC9), which is dimerized and is activated by the administration of an otherwise bioinert small-molecule drug, AP1903 (Spencer DM, etal. Controlling signal transduction with synthetic ligands., Science, 1993, vol. 2625136(pg. 1019-1024. Fan L, et al. Improved artificial death switches based on caspases and FADD., Hum Gene Ther, 1999, vol. 10 14, pg. 2273-2285. Straathof KC, et al. An inducible caspase 9 safety switch for T-cell therapy., Blood, 2005, vol. 105 11, pg. 4247-4254. Tey SK, et al. Induciblecaspase 9 suicide gene to improve the safety of allodepleted T cells after haploidentical stem cell transplantation., Biol Blood Marrow Transplant, 2007, vol. 13 8, pg. 913-924). Other examples are cytosine deaminase / 5-fluorocytosine and the herpes simplex virus / ganciclovir.[000160] Several delivery methods may be utilized in conjunction with the isolated nucleic acid sequences for in vitro (cell cultures) and in vivo (animals and patients) systems. In one embodiment, a lentiviral gene delivery system may be utilized. Such a system offers stable, longterm presence of the gene in dividing and non-dividing cells with broad tropism and the capacity for large DNA inserts. (Dull el al, J Virol, 72:8463-8471 1998) In an embodiment, adeno- associated virus (.AAV) may be utilized as a delivers- method. AAV is a non-pathogenic, single- stranded DNA virus that has been actively employed in recent years for delivering therapeutic gene in in vitro and in vivo systems (Choi et al, Citrr Gene Ther, 5:299-310, 2005). AAV include serotypes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, DJ or DJ / 8. An example of non-viral delivery method may utilize nanoparticle technology This platform has demonstrated utility as a pharmaceutical in vivo. Nanotechnology has improved transcytosis of drugs across tight epithelial and endothelial barriers. It offers targeted delivery of its payload to cells and tissues in a specific manner (Allen and CulHs, Science, 303: 1818-1822, 1998).[000161] The polynucleotides embodied herein may be used with a microdelivery vehicle such as cationic liposomes and adenoviral vectors. For a review of the procedures for liposome preparation, targeting and delivery- of contents, see Mannino and Gould-Fogerite, Bio Techniques, 6:682 (1988). See also, Feigner and Holm, Bethesda Res. Lab. Focus, 11(2):21 (1989) and Maurer, R. A., Bethesda Res. Lab. Focus, 1 1 (2): 25 (1989).[000162] Replication-defective recombinant adenoviral vectors can be produced in accordance with known techniques See, Quantin, el al., Proc. Natl. Acad. Sci. USA, 89:2581-2584 (1992), Stratford-Perricadet, et al., J. Clin. Invest., 90:626-630 (1992), and Rosenfeld, et al, Cell, 68: 143-155 (1992)[000163] Another method is to use single stranded DNA producing vectors which can produce the expressed products intracellularly. See for example, Chen et al, BioTechniques, 34: 167-171 (2003), which is incorporated herein, by reference, in its entirety.[000164] Th e nucleic acid sequences of the disclosure, e.g., vectors, can be delivered to an appropriate cell of a subject. This can be achieved by, for example, the use of a polymeric, biodegradable microparticle or microcapsule delivery vehicle, sized to optimize phagocytosis by phagocytic cells such as macrophages. For example, PLGA (poly-lacto-co-glycolide) microparticles approximately 1-10 pm in diameter can be used. The polynucleotide is encapsulated in these microparticles, which are taken up by macrophages and gradually biodegraded within the ceil, thereby releasing die polynucleotide. Once released, the DNA is expressed within the cell. A second type of microparticle is intended not to be taken up directly by cells, but rather to serve primarily as a slow-release reservoir of nucleic acid that is taken up by cells only upon release from the micro-particle through biodegradation. These polymeric particles should therefore be large enough to preclude phagocytosis (i.e., larger than 5 gm and preferably larger than 2.0 pm). Another way to achieve uptake of the nucleic acid is using liposomes, prepared by standard methods. The nucleic acids can be incorporated alone into these delivery7vehicles or coincorporated with cell- or tissue-specific antibodies, for example, specific for Treg cells or delivery to tumor cells as a target. Alternatively, one can prepare a molecular complex composed of a plasmid or other vector attached to poly-L-lysine by electrostatic or covalent forces. Poly-L-lysine binds to a ligand that can bind to a receptor on target cells Deliver,' of “naked DNA” (i e., without a deliver,' vehicle) to an intramuscular, intradermal, or subcutaneous site, is another means to achieve w vivo expression. In the relevant polynucleotides (e.g., expression vectors) the nucleic acid sequence encoding an isolated nucleic acid sequence comprising a sequence encoding a CCR, as described above.[000165] In some embodiments, the compositions of the disclosure can be formulated as a nanoparti de, for example, nanoparticles comprised of a core of high molecular weight linear polyethyleneimine (LPEI) complexed with DNA and surrounded by a shell of polyethylene glycol modified (PEGylated) low molecular weight LPEI. The nucleic acids and vectors may also be applied to a surface of a device (e.g., a catheter) or contained within a pump, patch, or other drug delivery' device. The nucleic acids and vectors disclosed herein can be administered alone, or in a mixture, in the presence of a pharmaceutically acceptable excipient or carrier (e.g , physiological saline). The excipient or carrier is selected on the basis of the mode and route of administration.Suitable pharmaceutical carriers, as well as pharmaceutical necessities for use in pharmaceutical formulations, are described in Remington's Pharmaceutical Sciences (E. W. Martin), a well-known reference text in this field, and in the USP / NF (United States Pharmacopeia and the National Formulary).[000166] In some embodiments, the compositions can be formulated as a nanoparticle encapsulating the compositions embodied herein.[000167] Regardless of whether compositions are administered as nucleic acids or polypeptides, they are formulated in such a way as to promote uptake by the mammalian cell. Useful vector systems and formulations are described above. In some embodiments the vector can deliver the compositions to a specific cell type. The disclosure is not so limited however, and other methods of DNA delivery such as chemical transfection, using, for example calcium phosphate, DEAE dextran, liposomes, lipoplexes, surfactants, and perfluoro chemical liquids are also contemplated, as are physical delivery methods, such as electroporation, micro injection, ballistic particles, and "‘gene gun” systems.[000168] Methods for Isolation of Cells[000169] Any number of methods known in the art can be used to isolate cells, such as NK cells, or any other cell type that may be used in carrying out the treatment of a subject. Thus, also provided are various other genetically engineered cells expressing the chimeric cytokine receptors (CCRs) embodied herein. The cells generally are eukaryotic cells, such as mammalian cells, and typically are human cells. In some embodiments, the cells are derived from the blood, bone marrow, lymph, or lymphoid organs, are cells of the immune system, such as cells of the innate or adaptive immunity, e g., myeloid or lymphoid cells, including lymphocytes, typically T cells and / or NK cells. Other exemplary’ cells include stem cells, such as multipoient and pluripotent stem cells, including induced pluripotent stem cells (iPSCs). The cells typically are primary cells, such as those isolated directly from a subject and / or isolated from a subject and frozen. In some embodiments, the cells include one or more NK cells and subpopulations thereof, such as those defined by function, activation state, maturity, potential for differentiation, expansion, recirculation, localization, and / or persistence capacities, antigen-specificity, type of antigenreceptor, presence in a particular organ or compartment, marker or cytokine secretion profile, and / or degree of differentiation. With reference to the subject to be treated, the cells may be allogeneic and / or autologous. In some aspects, the cells are pluripotent and / or multipotent, such as stem cells, such as induced pluripotent stem cells (iPSCs). In some embodiments, the methods include isolating cells from the subject, preparing, processing, culturing, and / or engineering them, as described herein, and re-introducing them into the same patient or different patient, before or after cry opre servati on .[000170] In some embodiments, the cells include one or more nucleic acids introduced via genetic engineering, and thereby express recombinant or genetically engineered products of such nucleic acids In some embodiments, the nucleic acids are heterologous, i.e., normally not present in a cell or sample obtained from the cell, such as one obtained from another organism or cell, which for example, is not ordinarily found in the cell being engineered and / or an organism from which such cell is derived. In some embodiments, the nucleic acids are not naturally occurring, such as a nucleic acid not found in nature, including one comprising chimeric combinations of nucleic acids encoding various domains from multiple different cell types.[000171] In some embodiments, preparation of the engineered cells includes one or more culture and / or preparation steps. The cells for introduction of the CCRs, may be isolated from a sample, such as a biological sample, e.g., one obtained from or derived from a subject. In some embodiments, the subject from which the cell is isolated is one having the disease or condition or in need of a cell therapy or to which cell therapy will be administered. The subject in some embodiments is a human in need of a particular therapeutic intervention, such as the adoptive cell therapy for which cells are being isolated, processed, and / or engineered.[000172] Accordingly, the cells in some embodiments are primary cells, e g., primary human cells. The samples include tissue, fluid, and other samples taken directly from the subject, as well as samples resulting from one or more processing steps, such as separation, centrifugation, genetic engineering (e g., transduction with viral vector), washing, and / or incubation. The biological sample can be a sample obtained directly from a biological source or a sample that is processed. Biological samples include, but are not limited to, body fluids, such as blood, plasma, serum,cerebrospinal fluid, synovial fluid, urine and sweat, tissue and organ samples, including processed samples derived therefrom.[000173] In some aspects, the sample from which the cells are derived or isolated is blood or a blood-derived sample or is derived from an apheresis or leukapheresis product. Exemplary samples include whole blood, peripheral blood mononuclear cells (PBMCs), brain, central nervous system (CNS), leukocytes, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut associated lymphoid tissue, mucosa associated lymphoid tissue, spleen, other lymphoid tissues, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testes, ovaries, tonsil, or other organ, and / or cells derived therefrom. Samples include, in the context of ceil therapy, e.g., adoptive cell therapy, samples from autologous and allogeneic sources.[000174] In some embodiments, the cells are derived from cell lines, e.g., NK cell lines. The cells in some embodiments are obtained from a xenogeneic source, for example, from mouse, rat, non-human primate, or pig.[000175] In some embodiments, isolation of the cells includes one or more preparation and / or non-affiniiy based cell separation steps. In some examples, cells are washed, centrifuged, and / or incubated in the presence of one or more reagents, for example, to remove unwanted components, enrich for desired components, lyse, or remove cells sensitive to particular reagents. In some examples, cells are separated based on one or more property, such as density, adherent properties, size, sensitivity and / or resistance to particular components[000176] In some examples, cells from the circulating blood of a subject are obtained, e g., by apheresis or leukapheresis. The samples, in some aspects, contain lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and / or platelets, and in some aspects contains cells other than red blood cells and platelets.[000177] In some embodiments, the blood cells collected from the subject are washed, e.g., io remove the plasma fraction and to place the cells in an appropriate buffer or media for subsequent processing steps In some embodiments, the cells are washed with phosphate buffered saline (PBS). In some embodiments, the wash solution lacks calcium and / or magnesium and / ormany or all divalent cations. In some aspects, a washing step is accomplished by a semi -automated “flow-through” centrifuge according to the manufacturer's instructions. In some aspects, a washing step is accomplished by tangential flow filtration (TFF) according to the manufacturer's instructions. In some embodiments, the cells are resuspended in a variety of biocompatible buffers after washing, such as, for example, Ca+7Mg4 rfree PBS In certain embodiments, components of a blood cell sample are removed, and the cells directly resuspended in culture media.[000178] In some embodiments, the methods include density-based cell separation methods, such as the preparation of white blood cells from peripheral blood by lysing the red blood cells and centrifugation through a Percoll or Ficoll gradient.[000179] In some embodiments, the isolation methods include the separation of different cell types based on the expression or presence in the cell of one or more specific molecules, such as surface markers, e.g., surface proteins, intracellular markers, or nucleic ac;d In some embodiments, any known method for separation based on such markers may be used. In some embodiments, the separation is affinity- or immunoaffinity-based separation. For example, the isolation in some aspects includes separation of cells and cell populations based on the cells' expression or expression level of one or more markers, typically cell surface markers, for example, by incubation with an antibody or binding partner that specifically binds to such markers, followed generally by washing steps and separation of cells having bound the antibody or binding partner, from those cells having not bound to the antibody or binding partner[000180] Such separation steps can be based on positive selection, in which the cells having bound the reagents are retained for further use, and / or negative selection, in which the cells having not bound to the antibody or binding partner are retained. In some examples, both fractions are retained for further use. In some aspects, negative selection can be particularly useful where no antibody is available that specifically identifies a cell type in a heterogeneous population, such that separation is best carried out based on markers expressed by ceils other than the desired population.[000181] Th e separation need not result in 100% enrichment or removal of a particular cell population or cells expressing a particular marker. For example, positive selection of or enrichment for cells of a particular type, such as those expressing a marker, refers to increasing the number orpercentage of such cells, but need not result in a complete absence of cel Is not expressing the marker. Likewise, negative selection, removal, or depletion of cells of a particular type, such as those expressing a marker, refers to decreasing the number or percentage of such cells, but need not result in a complete removal of all such cells.[000182] In some examples, multiple rounds of separation steps are carried out, where the positively or negatively selected fraction from one step is subjected to another separation step, such as a subsequent positive or negative selection. In some examples, a single separation step can deplete cells expressing multiple markers simultaneously, such as by incubating cells with a plurality of antibodies or binding partners, each specific for a marker targeted for negative selection. Likewise, multiple cell types can simultaneously be positively selected by incubating cells with a plurality of antibodies or binding partners expressed on the various cell types e.g. NK cells.[000183] In some embodiments, isolation is carried out by enrichment for a particular cell population by positive selection, or depletion of a particular ceH population, by negative selection. In some embodiments, positive or negative selection is accomplished by incubating cells with one or more antibodies or other binding agent that specifically bind to one or more surface markers expressed or expressed (markeriT’) at a relatively higher level (marker!,,gh) on the positively or negatively selected cells, respectively.[000184] In certain embodiments, the cells are NK cells wherein the NK cells are transduced with the CCRs, of the disclosure. NK cells have typically been identified in flow cytometry by first excluding other lymphocyte markers such as the CD3 T cell marker. After this sorting, two markers have become well-established as standard NK cell markers, CD56 (neural cell adhesion molecule-1, NCAM1) and CD16 (low affinity Fc gamma receptor 3A, FCGR3A, FcyRIII). The differential expression of these two surface proteins defines the two main subsets of conventional NK cells, CD56brighlCD16io" and CD56dimCD16\ often simplified as CD56brighland CD56din\ respectively. Among circulating cells in peripheral blood, CD56brigh’ is less abundant, estimated to comprise only 5- 10% of the population. CD56dimrepresents greater than 90% of NK cells. However, CD56br,gfahas been noted to be abundant in certain tissues, including secondary lymphoid tissues.[000185] In some aspects, the sample or composition of cells to be separated is incubated with small, magnetizable or magnetically responsive material, such as magnetically responsive particles or microparticles, such as paramagnetic beads (e.g., such as DYNABEADS or MACS beads). The magnetically responsive material, e.g., particle, generally is directly or indirectly attached to a binding partner, e.g., an antibody, that specifically binds to a molecule, e.g., surface marker, present on the cell, cells, or population of cells that it is desired to separate, e g., that it is desired to negatively or positively selected.[000186] In some embodiments, the magnetic particle or bead comprises a magnetically responsive material bound to a specific binding member, such as an antibody or other binding partner. There are many well-known magnetically responsive materials used in magnetic separation methods. Suitable magnetic particles include those described in Mol day, U.S. Pat. No. 4,452,773, and in European Patent Specification EP 452342 B, which are hereby incorporated by reference Colloidal sized particles, such as those described in Owen U.S. Pat, No 4,795,698, and Liberti eta!., U.S. Pat. No. 5,200,084 are other examples.[000187] The incubation generally is carried out under conditions whereby the antibodies or binding partners, or molecules, such as secondary antibodies or other reagents, which specifically bind to such antibodies or binding partners, which are attached to the magnetic particle or bead, specifically bind to cell surface molecules if present on cells within the sample.[000188] In some aspects, the sample is placed in a magnetic field, and those cells having magnetically responsive or magnetizable particles attached thereto will be attracted to the magnet and separated from the unlabeled cells. For positive selection, cells that are attracted to the magnet are retained; for negative selection, cells that are not attracted (unlabeled cells) are retained. In some aspects, a combination of positive and negative selection is performed during the same selection step, where the positive and negative fractions are retained and further processed or subject to further separation steps[000189] In certain embodiments, the magnetically responsive particles are coated in primary antibodies or other binding partners, secondary antibodies, lectins, enzymes, or streptavidin. In certain embodiments, the magnetic particles are attached to cells via a coating of primaryantibodies specific for one or more markers. In certain embodiments, the cells, rather than the beads, are labeled with a primary antibody or binding partner, and then cell-type specific secondary antibody- or other binding partner (e.g , streptavidin)-coated magnetic particles, are added. In certain embodiments, streptavidin-coated magnetic particles are used in conjunction with biotinylated primary or secondary antibodies.[000190] In some embodiments, the magnetically responsive particles are left attached to the cells that are to be subsequently incubated, cultured and / or engineered; in some aspects, the particles are left attached to the cells for administration to a patient In some embodiments, the magnetizable or magnetically responsive particles are removed from the ceils. Methods for removing magnetizable particles from cells are known and include, e.g , the use of competing nonlabeled antibodies, magnetizable particles or antibodies conjugated to cleavable linkers, etc. In some embodiments, the magnetizable particles are biodegradable.[000191] In some embodiments, the affinity-based selection is via magnetic-activated cell sorting (MACS) (Miltenyi Biotec, Auburn, Calif ). Magnetic Activated Cell Sorting (MACS) systems are capable of high-purity selection of cells having magnetized particles attached thereto. In certain embodiments, MACS operates in a mode wherein the non-target and target species are sequentially eluted after the application of the external magnetic field. That is, the cells attached to magnetized particles are held in place while the unattached species are eluted. Then, after this first elution step is completed, the species that were trapped in the magnetic field and were prevented from being eluted are freed in some manner such that they can be eluted and recovered In certain embodiments, the non-target cells are labelled and depleted from the heterogeneous population of cells[000192] In certain embodiments, the isolation or separation is carried out using a system, device, or apparatus that carries out one or more of the isolation, cell preparation, separation, processing, incubation, culture, and / or formulation steps of the methods. In some aspects, the system is used to carry out each of these steps in a closed or sterile environment, for example, to minimize error, user handling and / or contamination.[000193] In some embodiments, the system or apparatus carries out one or more, e g., all, of the isolation, processing, engineering, and formulation steps in an integrated or self-contained system, and / or in an automated or programmable fashion. In some aspects, the system or apparatus includes a computer and / or computer program in communication with the system or apparatus, which allows a user to program, control, assess the outcome of, and / or adjust various aspects of the processing, isolation, engineering, and formulation steps.[000194] In some aspects, the separation and / or other steps is carried out using CliniMACS system (Miltenyi Biotec), for example, for automated separation of cells on a clinical-scale level in a closed and sterile system. Components can include an integrated microcomputer, magnetic separation unit, peristaltic pump, and various pinch valves. The integrated computer in some aspects controls all components of the instrument and directs the system to perform repeated procedures in a standardized sequence. The magnetic separation unit in some aspects includes a movable permanent magnet and a holder for the selection column. The peristaltic pump controls the flow rate throughout the tubing set and, together with the pinch valves, ensures the controlled flow of buffer through the system and continual suspension of cells.[000195] Th e CliniMACS system in some aspects uses antibody-coupled magnetizable particles that are supplied in a sterile, non-pyrogenic solution. In some embodiments, after labelling of cells with magnetic particles the cells are washed to remove excess particles. A cell preparation bag is then connected to the tubing set, which in turn is connected to a bag containing buffer and a cell collection bag. The tubing set consists of pre-assembled sterile tubing, including a pre-column and a separation column, and are for single use only. After initiation of the separation program, the system automatically applies the cell sample onto the separation column Labelled cells are retained within the column, while unlabeled cells are removed by a series of washing steps. In some embodiments, the cell populations for use with the methods described herein are unlabeled and are not retained in the column. In some embodiments, the cell populations for use with the methods described herein are labeled and are retained in the column. In some embodiments, the cell populations for use with the methods described herein are eluted from the column after removal of the magnetic field and are collected within the cell collection bag.[000196] In certain embodiments, separation and / or other steps are carried out using the CliniMACS Prodigy system (Miltenyi Biotec). The CliniMACS Prodigy system in some aspects is equipped with a cell processing unity that permits automated washing and fractionation of ceils by centrifugation. The CliniMACS Prodigy system can also include an onboard camera and image recognition software that determines the optimal cell fractionation endpoint by discerning the macroscopic layers of the source cell product. For example, peripheral blood may be automatically separated into erythrocytes, white blood cells and plasma layers. The CliniMACS Prodigy system can also include an integrated cell cultivation chamber which accomplishes cell culture protocols such as, e.g., cell differentiation and expansion, antigen loading, and long-term cell culture. Input ports can allow for the sterile removal and replenishment of media and cells can be monitored using an integrated microscope. See, e.g., Klebanoff et al. (2012) J Imtmmoiher, 35(9): 651-660, Terakura et al. (2012) Blood. 1:72-82, and Wang et al. (2012) Innmtnather. 35(9):689-70l .[000197] In some embodiments, a cell population described herein is collected and enriched (or depleted) via flow cytometry, in which cells stained for multiple cell surface markers are carried in a fluidic stream. In some embodiments, a cell population described herein is collected and enriched (or depleted) via preparative scale (FACS)-sorting. In certain embodiments, a cell population described herein is collected and enriched (or depleted) by use of microelectromechanical systems (MEMS) chips in combination with a FACS-based detection system (see, e.g., WO 2010 / 033140, Cho et al. (2010) Lab Chip 10, 1567-1573, and Godin et al. (2008) J Biophoton. l(5):355-376. In both cases, cells can be labeled with multiple markers, allowing for the isolation of well-defined cell subsets at high purity.[000198] In some embodiments, the antibodies or binding partners are labeled with one or more detectable markers, to facilitate separation for positive and / or negative selection. For example, separation may be based on binding to fluorescently labeled antibodies. In some examples, separation of cells based on binding of antibodies or other binding partners specific for one or more cell surface markers are carried in a fluidic stream, such as by fluorescence-activated cell sorting (FACS), including preparative scale (FACS) and / or microelectromechanical systems (MEMS) chips, e.g., in combination with a flow-cytometric detection system. Such methods allow for positive and negative selection based on multiple markers simultaneously.[000199] In some embodiments, the preparation methods include steps for freezing, e g., cryopreserving, the cells, either before or after isolation, incubation, and / or engineering. In some embodiments, the freeze and subsequent thaw step removes granulocytes and, to some extent, monocytes in the cell population. In some embodiments, the cells are suspended in a freezing solution, e.g.. following a washing step to remove plasma and platelets. Any of a variety of known freezing solutions and parameters in some aspects may be used. One example involves using PBS containing 20% DMSO and 8% human serum albumin (FISA), or other suitable cell freezing media. This is then diluted 1: 1 with media so that the final concentration of DMSO and HSA are10% and 4%, respectively. The cells are then frozen to -80° C at a rate of 1°C per minute and stored in the vapor phase of a liquid nitrogen storage tank.[000200] In some embodiments, the provided methods include cultivation, incubation, culture, and / or genetic engineering steps. For example, in some embodiments, provided are methods for incubating and / or engineering the depleted cell populations and culture-initiating compositions.[000201] Thus, in some embodiments, the cell populations are incubated in a cultureinitiating composition The incubation and / or engineering may be carried out in a culture vessel, such as a unit, chamber, well, column, tube, tubing set, valve, vial, culture dish, bag, or other container for culture or cultivating ceils.[000202] In some embodiments, the cells are incubated and / or cultured prior to or in connection with genetic engineering. The incubation steps can include culture, cultivation, stimulation, activation, and / or propagation. In some embodiments, the compositions or cells are incubated in the presence of stimulating conditions or a stimulatory agent. Such conditions include those designed to induce proliferation, expansion, activation, and / or survival of cells in the population, to mimic antigen exposure, and / or to prime the cells for genetic engineering, such as for the introduction of a recombinant antigen receptor.[000203] Th e conditions can include one or more of particular media, temperature, oxygen content, carbon dioxide content, time, agents, e.g., nutrients, amino acids, antibiotics, ions, and / orstimulatory factors, such as cytokines, chemokines, antigens, binding partners, fusion proteins, recombinant soluble receptors, and any other agents designed to activate the cells.[000204] Methods of Treatment[000205] In certain embodiments, a method of treating a subject requiring imrmmotherapy such as for exampie, cancer, and the like comprises isolating NK cells from a biological sample obtained from the subject; transducing the NK cells with an expression vector encoding a CCR embodied herein, which specifically binds to, for example CD 123; stimulating the transduced NK cells with a CCR embodied herein, at least once ex vivo; and reinfusing the cells into the subject, thereby treating the subject. In certain embodiments, the cells are allogeneic cells. In certain embodiments, the cells are autologous cells. The NK cells may be generated from any suitable source ofNK cells known in the art including, but not limited to, NK cells collected from a subject The subject may be a patient with cancer in need of NK cell therapy or a subject of the same species as the subject with the disease in need of cell therapy. The collected cells may be expanded ex vivo using methods commonly known in the art before transduction with a CCR embodied herein to generate CCR bearing NK cells.[000206] The CCR NK cells, once they have been expanded ex vivo in response to, for example, a cancer antigen, can be reinfused into the subject in a therapeutically effective amount The term “therapeutically effective amount’' as used herein means the amount of CCR NK cells when administered to a mammal, in particular a human, in need of such treatment, is sufficient to treat cancer, virus infections, etc. In certain embodiments, administration of any of the compositions embodied herein, can be combined with other cell-based therapies, for example, stem cells, antigen presenting cells, pancreatic islets etc.[000207] The composition of the present disclosure may be prepared in a manner known in the art and in a manner suitable for parenteral administration to mammals, particularly humans, comprising a therapeutically effective amount of the composition alone, with one or more pharmaceutically acceptable carriers or diluents.[000208] I he term “pharmaceutically acceptable carried' as used herein means any suitable carriers, diluents or excipients. These include ail aqueous and non-aqueous isotonic sterileinjection solutions which may contain anti-oxidants, buffers and solutes, which render the composition isotonic with the blood of the intended recipient; aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents, dispersion media, antifungal and antibacterial agents, isotonic and absorption agents and the like. It will be understood that compositions of the invention may also include other supplementary physiologically active agents.[000209J The carrier must be pharmaceutically "acceptable” in the sense of being compatible with the other ingredients of die composition and not injurious to the subject. Compositions include those suitable for parenteral administration, including subcutaneous, intramuscular, intravenous and intradermal administration. The compositions may conveniently be presented in unit dosage form and may be prepared by any method well known in the art of pharmacy. Such methods include preparing the earner for association with the NK cells expressing CCRs. In general, the compositions are prepared by uniformly and intimately bringing into association any active ingredients with liquid carriers.[000210] In an embodiment, the composition is suitable for parenteral administration. In another embodiment, the composition is suitable for intravenous administration.[000211] Compositions suitable for parenteral administration include aqueous and nonaqueous isotonic sterile injection solutions which may contain anti-oxidants, buffers, bactericides and solutes, which render the composition Isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.[000212] In other embodiments, the compositions comprise a cell which has been transformed or transfected with one or more vectors or nucleic acids encoding one or more CCR. In some embodiments, the methods of the disclosure can be applied ex vivo. That is, a subject's cells can be removed from the body and transduced with the compositions in culture with a desired target antigen NK cells returned to the subject's body The cells can be irradiated to prevent replication. In some embodiments, the cells are human leukocyte antigen (HLA)-rnatched, autologous, cell lines, or combinations thereof. In other embodiments the cells can be a stem cell.For example, an embryonic stem cel 1 or an artificial pluripotent stem cell (induced pluripotent stem cell (iPS cell)). Embryonic stem cells (ES cells) and artificial pluripotent stem cells (induced pluripotent stem cell. iPS cells) have been established from many animal species, including humans. These types of pluripotent stem ceils would be the most useful source of cells for regenerative medicine because these cells are capable of differentiation into almost all of the organs by appropriate induction of their differentiation, with retaining their ability of actively dividing while maintaining their pluripotency. IPS cells, in particular, can be established from selfderived somatic cells, and therefore are not likely to cause ethical and social issues, in comparison with ES cells which are produced by destruction of embryos. Further, IPS cells, which are selfderived cell, make it. possible to avoid rejection reactions, which are the biggest obstacle to regenerative medicine or transplantation therapy.[000213] The CCR complexes can be easily delivered io a subject by methods known in the art, for example, methods which deliver siRNA Thus, the CCR complexes can be used clinically, similar to the approaches taken by current gene therapy. In particular, a CAR stable expression stem cell or iPS cells for cell transplantation therapy as well as vaccination can be developed for use in subjects.[000214] The NK cells expressing the CCRs embodied herein, once they have been expanded ex vivo etc., are reinfused into the subject in a therapeutically effective amount. The term “therapeutically effective amount” as used herein means the amount of NK cells expressing the CCRs embodied herein when administered to a mammal, in particular a human, in need of such treatment, is sufficient to treat the disease.[000215] The precise amount of NK cells expressing the CCRs embodied herein to be administered can be determined by a physician with consideration of individual differences in age, weight, extent of disease and condition of the subject. Typically, administration of NK cell therapies is defined by number of cells per kilogram of body weight. However, because NK cells will replicate and expand after transfer, the administered cell dose will not resemble the final steady-state number of cells.[000216] In an embodiment, a pharmaceutical composition comprising the NK cells expressing the CCRs embodied herein of the present disclosure may be administered at a dosage of 104to 109cells / kg body weight. In another embodiment, a pharmaceutical composition comprising the NK cells expressing the CCRs embodied herein of the present disclosure may be administered at a dosage of 105to IO6cells / kg body weight, including all integer values within those ranges.[000217] Compositions comprising the NK cells expressing the CCRs embodied herein of the present disclosure may also be administered multiple times at these dosages. The cells can be administered by using infusion techniques that are known in the art (see, for example, Rosenberg el al., 1988, Afw Englana ’ Journal of Medicine, 319: 1676). The optimal dosage and treatment regimen for a particular subject can be readily determined by one skilled in the art by monitoring the patient for signs of disease and adjusting the treatment accordingly.[000218] COMHiNAI ION THERAPIES[000219] The disclosure also contemplates the combination of the composition of the present disclosure with other drugs and / or in addition to other treatment regimens or modalities such as surgery. When the composition of the present disclosure is used in combination with known therapeutic agents the combination may be administered either in sequence (either continuously or broken up by periods of no treatment) or concurrently or as an admixture. For example, in the case of cancer chemotherapeutic agents may be administered as part of the combination therapy.[000220] In certain embodiments, the NK cells expressing the CCRs embodied herein are administered in conjunction with a cancer therapy As used herein, the term “cancer therapy” refers to a therapy useful in treating cancer. Examples of anti-cancer therapeutic agents include, but are not limited to, e.g., surgery, chemotherapeutic agents, immunotherapy, growth inhibitory agents, cytotoxic agents, agents used in radiation therapy, anti-angiogenesis agents, apoptotic agents, antitubulin agents, and other agents to treat cancer, such as anti-HER-2 antibodies (e.g., HERCEPTIN™), anti-CD20 antibodies, an epidermal growth factor receptor (EGFR) antagonist (e.g., a tyrosine kinase inhibitor), HER1 / EGFR inhibitor (e.g., erlotinib (TARCEVA™)), platelet derived growth factor inhibitors (e.g., GLEEVEC™ (Imatinib Mesylate)), a COX-2 inhibitor (e.g.,celecoxib), interferons, cytokines, antagonists (e g., neutralizing antibodies) that bind to one or more of the following targets ErbB2, ErbB3, ErbB4, PDGFR-beta, BlyS, APRIL, BCMA or VEGF receptor(s), TRAIL / Apo2, and other bioactive and organic chemical agents, etc. Combinations thereof are also contemplated for use with the methods described herein.[000221] A “chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include Erlotinib (TARCEVA™, Genentech / OSI Pharm.), Bortezomib (VELCADE™, Millennium Pharm.), Fulvestrant (FASLODEX™, Astrazeneca), Sutent (SU11248, Pfizer), Letrozole (FEMARA™, Novartis), Imatinib mesylate (GLEEVEC™, Novartis), PTK787 / ZK 222584 (Novartis), Oxaliplatin (Eloxatin™, Sanofi), 5-FU (5-fluorouracil), Leucovorin, Rapamycin (Sirolimus, RAPAMUNE™, Wyeth), Lapatinib (GSK572016, GlaxoSmithKline), Lonafarnib (SCH 66336), Sorafenib (BAY43-9006, Bayer Labs.), and Gefitinib (IRESSA™, Astrazeneca), AG1478, AG1571 (SU 5271; Sugen), alkylating agents such as Thiotepa and CYTOXAN1Mcyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozcicsin, carzcicsin and bizcicsin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pan crati statin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin yl and calicheamicin omega 1 (Angew Chem. Inti. Ed. Engl. (1994) 33:183-186); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, anthramycin, azaserine,bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN™ doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, strcptonigrin, strcptozocin, tubcrcidin, ubenimcx, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5 -fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6- mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacytidine, 6- azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK™ polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosinc; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g., TAXOL™ paclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE1MCremophor-free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE™ doxetaxel (Rhone-Poulenc Rorer, Antony, France); chloranbucil; GEMZAR™ gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE™ vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RES 2000; difluoromethylornithine(DMFO); retinoids such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above.[000222] Also included in this definition of “chemotherapeutic agent” are: (i) anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX™ (tamoxifen)), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and FARESTON™ (toremifene); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, MEGASE™ (megestrol acetate), AROMASIN™ (exemestane), formestanie, fadrozole, RIVISOR™ (vorozole), FEMARA™ (letrozole), and ARIMIDEX™ (anastrozole); (iii) anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; as well as troxacitabine (a 1,3-di oxolane nucleoside cytosine analog); (iv) aromatase inhibitors; (v) protein kinase inhibitors; (vi) lipid kinase inhibitors; (vii) antisense oligonucleotides, particularly those which inhibit expression of genes in signaling pathways implicated in aberrant cell proliferation, such as, for example, PKC-alpha, Ralf and H-Ras; (viii) ribozymes such as a VEGF expression inhibitor (e.g., ANGIOZYME™ (ribozyme)) and a HERZ expression inhibitor; (ix) vaccines such as gene therapy vaccines, for example, ALLOVECTIN™ vaccine, LEUVECTIN™ vaccine, and VAXID™ vaccine; PROLEUKIN™ rIL-2; LURTOTECAN™ topoisomerase 1 inhibitor; ABARELIX™ rmRH; (x) anti-angiogenic agents such as bevacizumab (AVASTIN™, Genentech); and (xi) pharmaceutically acceptable salts, acids or derivatives of any of the above.[000223] In various embodiments, the cancer therapeutic is an immunotherapy selected from the group comprising oncolytic virus, bacteria, oncolytic bacteria or other bacterial compositions, Bacillus Calmette-Guerin (BCG), a microbiome modulator, and / or a toll-like receptor (TLR) agonist. In various embodiments, the TLR agonist is a TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, TLR10, TLR11, TLR12, and / or TLR13 agonist. In various embodiments, the TLR agonist is derived from virus, plants, bacteria and / or made synthetically. In various embodiments, the immunotherapy is a is a stimulator of interferon genes (STING) pathway modulator.[000224] It will be appreciated by those skilled in the art of cancer immunotherapy that other complementary immune therapies may be added to the regimens described above to further enhance their efficacy including but not limited to GM-CSF to increase the number of myeloid derived innate immune system cells, low dose cyclophosphamide or PI3K inhibitors (e.g., PI3K delta inhibitors) to eliminate T regulatory cells that inhibit innate and adaptive immunity and 5FU (e.g., capecitabine), PI3K inhibitors or histone deacetylase inhibitors to remove inhibitory myeloid derived suppressor cells. For example, PI3K inhibitors include, but are not limited to, LY294002, Perifosine, BKM120, Duvelisib, PX-866, BAY 80-6946, BEZ235, SF1126, GDC-0941, XL147, XL765, Palomid 529, GSK1059615, PWT33597, IC87114, TGI 00-15, CAL263, PI-103, GNE- 477, CUDC-907, and AEZS-136. In some aspects, the PI3K inhibitor is aPI3K delta inhibitor such as, but not limited to, Idelalisib, RP6530, TGR1202, and RP6503. Additional PI3K inhibitors are disclosed in U.S. Patent Application Nos. US20150291595, US20110190319, and International Patent Application Nos. WO2012146667, WO2014164942, WO2012062748, and WO2015082376. The immunotherapy may also comprise the administration of an interleukin such as IL-2, or an interferon such as INFa.[000225] In certain embodiments, the NK cells expressing the CCRs embodied herein are administered with one or more immune checkpoint modulators. Immune checkpoints refer to inhibitory pathways of the immune system that are responsible for maintaining self-tolerance and modulating the duration and amplitude of physiological immune responses. Examples of checkpoint inhibitor include, without limitation an inhibitor of: PD-1, PD-L1, PD-L2, CTLA4, TIM-3, LAG-3, CEACAM-1, CEACAM-5, VISTA, BTLA, TIGIT, LAIR1, CD 160, 2B4 or TGFR-p.[000226] The term “checkpoint inhibitor” means a group of molecules on the cell surface of CD4+and / or CD8+T cells that fine-tune immune responses by down-modulating or inhibiting an anti-tumor immune response. Immune checkpoint proteins are well known in the art and include, without limitation, CTLA-4, PD-1, VISTA, B7-H2, B7-H3, PD-L1, B7-H4, B7-H6, 2B4, ICOS, HVEM, PD-L2, CD160, gp49B, PIR-B, KIR family receptors, TIM-1, TIM-3, TIM-4, LAG-3, BTLA, SIRPa (CD47), CD48, 2B4 (CD244), B7.1, B7.2, ILT-2, ILT-4, TIGIT, and A2aR (see, for example, WO 2012 / 177624). “Anti-immune checkpoint inhibitor therapy” refers to the use ofagents that inhibit immune checkpoint inhibitors. Inhibition of one or more immune checkpoint inhibitors can block or otherwise neutralize inhibitory signaling to thereby upregulate an immune response in order to more efficaciously treat cancer. Exemplary agents useful for inhibiting immune checkpoint inhibitors include antibodies, small molecules, peptides, peptidomimetics, natural ligands, and derivatives of natural ligands, that can either bind and / or inactivate or inhibit immune checkpoint proteins, or fragments thereof; as well as RNA interference, antisense, nucleic acid aptamers, etc. that can downregulate the expression and / or activity of immune checkpoint inhibitor nucleic acids, or fragments thereof. Exemplary agents for upregulating an immune response include antibodies against one or more immune checkpoint inhibitor proteins block the interaction between the proteins and its natural receptor(s); a non-activating form of one or more immune checkpoint inhibitor proteins (e.g., a dominant negative polypeptide); small molecules or peptides that block the interaction between one or more immune checkpoint inhibitor proteins and its natural receptor(s); fusion proteins (e.g. the extracellular portion of an immune checkpoint inhibition protein fused to the Fc portion of an antibody or immunoglobulin) that bind to its natural receptor(s); nucleic acid molecules that block immune checkpoint inhibitor nucleic acid transcription or translation; and the like. Such agents can directly block the interaction between the one or more immune checkpoint inhibitors and its natural receptor(s) (e.g., antibodies) to prevent inhibitory signaling and upregulate an immune response. Alternatively, agents can indirectly block the interaction between one or more immune checkpoint proteins and its natural receptor(s) to prevent inhibitory signaling and upregulate an immune response. For example, a soluble version of an immune checkpoint protein ligand such as a stabilized extracellular domain can binding to its receptor to indirectly reduce the effective concentration of the receptor to bind to an appropriate ligand. In one embodiment, anti-PD-1 antibodies, anti-PD-Ll antibodies, and anti-CTLA-4 antibodies, either alone or used in combination.[000227] In some embodiments, such therapy involves blockade of programmed cell death 1 (PD-1). In some embodiments, such therapy involves treatment with an agent that interferes with an interaction involving PD-1 (e.g., with PD-L1). In some embodiments, such therapy involves administration of an antibody agent that specifically interacts with PD-1 or with PD-L1. In some embodiments, such therapy involves administration of one or more of nivolumab (BMS-936558,MDX-1106, ONO-4538, a fully human Immunoglobulin G4 (IgG4) monoclonal PD-1 antibody), pembrolizumab (MK-3475, a humanized monoclonal IgG4 anti-PD-1 antibody), BMS-936559 (a fully human IgG4 PD-L1 antibody), MPDL3280A (a humanized engineered IgGl monoclonal PD- L1 antibody) and / or MEDI4736 (a humanized engineered IgGl monoclonal PD-L1 antibody).[000228] Each embodiment disclosed herein is contemplated as being applicable to each of the other disclosed embodiments. Thus, all combinations of the various elements described herein are within the scope of the disclosure.[000229] This disclosure is further illustrated by the following examples which should not be construed as limiting. The contents of all references, patents, and published patent applications cited throughout this application, as well as the figures and the sequence listing, are hereby incorporated by reference.EXAMPLES[000230] EXAMPLE 1: CHIMERIC CYTOKINE RECEPTORS INDUCE ANTIGEN-SPECIFIC ACTIVATION IN NK CELLS AGAINST AML[000231] Chimeric antigen receptor (CAR) T cell therapies have revolutionized the standard of care treatment for B cell malignancies. However, CAR-T cell clinical trials for acute myeloid leukemia (AML) have had discouraging outcomes. As an alternative effector to T cells, natural killer (NK) cells are being investigated. NK cells are innately cytotoxic against cancer cells, are readily enhanced with CARs, and, unlike T cells, do not cause graft-versus-host disease. However, NK cells have exhibited a lack of persistence in multiple clinical trials, limiting their efficacy and the relapse-free survival of patients.[000232] Methods[000233] Artificial receptors were designed with AlphaFold using sequences from either the native IL-15R0 chain or the common gamma chain (yc) and single chain variable fragments to target CD 123, an AML-associated antigen. Each CCR chain was subcloned into an expression vector with a fluorescent protein, GFP or m Scarlet (mSc), to create various CCR-yc-GFP and CCR-IL-15Rp-mSc constructs. Primary NK cells were expanded from peripheral bloodmononuclear cells with irradiated K562.mbILl 5.41BBL feeder cells following magnetic depletion of T cells. NK cells were then transduced with retroviral vectors. After testing constructs with promising predicted in silico folding, the optimal CCR-pair was chosen for further testing based on surface expression and antigen-binding. Antigen-specific signaling of CCRs was evaluated by stimulating CCR-NK cells with immobilized recombinant human CD123 (rhCD123) and detecting activation of STAT5 by phosphorylation (pSTAT5) using Western blot. Antigen-specific cytotoxicity was assessed in 18-hour bioluminescence assays with firefly luciferase-expressing target cell lines. Non-transduced NK cells were used as controls in all experiments.[000234] Results[000235] CCR chains were expressed at similar levels individually (mean ± SD; CCR-yc- GFP, GFP%+: 66.3 ± 18.2%; CCR-IL-15RP-mSc, mSc+%: 64.5 ± 16.3%) and together (cotransduced, GFP+mSc+%: 53.2 ± 13.3). CCR single chain pSTAT5 was not detectable with rhCD123 stimulation and while NK cells co- transduced with both receptors displayed tonic signaling, stimulation with rhCD123 resulted in a 2-fold increase in signal. Co-transduced NK cells displayed significantly higher cytotoxicity versus MOLM-13 (CD123+) compared to NKs expressing single CCRs and non-transduced cells (1 :2 p <0.0001, 1 :1 p <0.0001 vs. nontransduced). NK cells did not show different cytotoxicity against Raji (CD123') (all p > 0.6691 vs. non-transduced).[000236] Conclusions[000237] NK cells expressing the engineered IL-15R-based CCRs demonstrate antigenspecific signaling and cytotoxicity dependent on expression of both our chimeric IL-15RP and chimeric yc.[000238] EXAMPLE 2: CHIMERIC CYTOKINE RECEPTORS TO EXTEND NATURAL KILLER CELL PERSISTENCE AND ENHANCE ANTI-AML EFFICACY[000239] In this study, it is sought to develop an alternate mechanism of CAR-NK cell stimulation that mimics the beneficial intracellular signaling of the IL- 15 receptor without unspecific systemic toxicity to achieve persistence of adoptively transferred NK cells.[000240] Antigen-specific cytokine signaling will be induced using chimeric cytokine receptors (CCRs). While activation domains in CARs designed for T and NK cell expression have been derived from co-stimulatory molecules (e.g., CD3(^, CD28, 2B4, 4-1BB),13the plan is to utilize domains from the common cytokine receptor gamma (yc) family (FIG. 1A). Antigenspecific cytokine signaling was engineered by linking a single-chain variable fragment (scFv) specific for the AML-associated antigen CD 123 (IL-3Ra) to the transmembrane and intracellular domains of the yc and IL-15RP chains (CCR-yc and CCR-IL-15RP, respectively), as yc family receptors function as heterodimers of the yc chain and an interleukin-specific chain (FIG. IB).14These two receptor chains were expressed in NK cells.[000241] Without wishing to be bound by theory, it was hypothesized that CCRs with antigen-specific cytokine signaling will increase NK persistence and confer powerful long-term anti-AML efficacy while avoiding in vivo toxicity. By utilizing heterodimerized complexes in the same fashion as native cytokine signaling, the aim is to surmount current clinical limitations of NK cell therapies. Also, using study of engineered CCRs, will ultimately lead to better understanding of the contribution of cytokine receptor-dependent synapse formation to the biology of NK cell activation.[000242] Cytokine treatments have been used to enhance the anti-cancer potential of immune cells engineered with CARs.17Additionally, switch cytokine receptors using extracellular domains from one cytokine receptor linked to intracellular signaling domains from others have been described and used in the development of T cell immunotherapies.18However, tumor-antigen- specific cytokine signaling has not been engineered into immune cells. Although cytokine signaling can enhance NK cell activity, other groups have relied on CARs for cytotoxic activity and it has been unclear if cytokine signaling alone in the context of an immunological synapse will stimulate NK cells enough to effectively kill targets. It is expected that the cytokine signal transduction, with conditional activation dependent on antigen-specific interactions, will eliminate the systemic toxicities observed with soluble cytokine supplements.[000243] RESULTS[000244] To optimize the expression of the CCRs, in silica modeling was used to predict the folding of CCR-yc constructs with shortening lengths of the native yc extracellular (FIG. 1C). Two CCR-yc constructs were chosen for in vitro testing based on this modeling and three CCR- IL-15RP were designed (FIG. ID). Two different scFvs (26292 or 32716)21are used for each chain in a CCR-pair to encourage epitope binding on the same CD123 molecule. The initial studies utilized CCR chains fused to fluorescent proteins for tracking. The sequences of these CCR constructs were synthesized and validated. The expression and antigen binding of the constructs of various lengths was evaluated using flow cytometry and his-tagged recombinant human CD 123 (rhCD123) staining (FIG. 2). Higher antigen binding and greater overall expression was observed when more of the native receptor chain extracellular domain was included in each of the chimeric chains. Thus, the longest CCR-yc and CCR-IL-15RP constructs (FIG. 3A, 3D and the following Tables 1 and 1A (Table 1 also shown in FIG. 3D) were chosen for further evaluation.Table 1A. Fluorescent protein tag labeled CCR DNA and protein sequences. DNA sequences of CCRs chosen for further development. Protein sequences are annotated based on the legend on the lower right. CCRs were encoded on two separate plasmids and were labeled with encoded fluorescent protein tags. The pSFG backbone was used for both plasmids.Table 1A. Fluorescent protein tag labeled CCR DNA and protein sequences (without stop codons). DNA sequences of CCRs chosen for further development. Protein sequences are annotated based on the legend on the lower left. CCRs were encoded on two separate plasmids and were labeled with encoded fluorescent protein tags. The pSFG backbone was used for both plasmids. These sequences of Table 1A correspond to those of Table 1 above except the stop sequence at the sequence end is removed. The signal sequence is cleaved during protein processing and is not present in the final construct. Protein sequences are annotated based on the legend on the lower right.TABLE 2Table 2. CCR DNA and protein sequences. To enable more efficient viral production and transduction, chosen chimeric yc and IL-15R0 sequences were subcloned into a single pMSGV expression vector and separated by a viral T2A skip sequence. Individual constructs were subcloned into pMSGV plasmids to serve as controls. Fluorescent protein tags were excluded from these vectors. Protein sequences are annotated based on the legend on the lower right.TABLE 3Table 3. Anti-B7H3 CCR DNA and protein sequences. Anti-CD123 scFvs on chimeric yc and IL-15R0 sequences were replaced with sequences targeting B7H3 (CD276). Validated anti-B7H3 scFv sequences (MGA271 and humanized 8H9) were used. Protein sequences are annotated based on the legend on the lower right.[000245] The transduction efficiency, cytotoxicity, and antigen-specific signaling of NK cells transduced with the best expressing CCR-yc, CCR-IL-15RP, and CCR-yc + CCR-IL-15RP were compared. Non-transduced NK cells served as controls in this study. Monocistronic vectors were co-transduced into NK cells to express CCR-yc + CCR-IL-15Rp. These constructs were stably integrated and expressed in primary human NK cells (FIGS. 3B, 3C). Transduction was measured by fluorescent tag expression (mean ± SD; CCR-yc-GFP, GFP%+: 66.3 ± 18.2%; CCR- IL-15RP-mSc, mSc+%: 64.5 ± 16.3%); co-transduced, GFP+mSc+%: 53.2 ± 13.3%). The ability of these constructs to bind the intended target was evaluated with his-tagged rhCD123 staining (mean ± SD, anti-his+%; CCR-yc-GFP: 34.9 ± 8.7%; CCR-IL-15Rp-mSc: 37.4 ± 12.1%; cotransduced: 42.4 ± 7.2%). Additionally, surface expression, as compared to total expression, was calculated by staining cells with an anti-mouse IgG flow antibody (mean ± SD, anti-F(ab)2+%; CCR-yc-GFP: 37.5 ± 14.6%; CCR-IL-15RP-mSc: 63.1 ± 12.6%; co-transduced: 60.0 ± 10.5%), as the 26292 and 32716 scFvs are derived from murine sequences.[000246] Example 3: Methodology[000247] NK cells engage other cells in an immunologic synapse. If a cell sends activating signals (e.g., stress-induced signals from viruses or DNA damage) that overwhelm inhibitory signals (e.g., presence of MHC class I), an NK cell will kill the engaged cell.13- 19, 20Cytokine signaling can enhance NK cell targeted cytotoxicity. However, cytokine signaling has the potential to cause inflammatory toxicity if not regulated. CCRs will allow NK cells to synapse with target cells to induce beneficial cytokine signaling that will direct NK cells to kill bound targets. The data herein demonstrates that antigen-specific IL- 15 receptor signaling alone (yc + IL-15RP) is enough to induce cytotoxicity. The aim is to evaluate NK cell CCR-dependent persistence and anti-tumor cytotoxicity as well as general safety.[000248] Plasmid construction. The CCR-IL-15RP + CCR-yc pair was subcloned into retroviral vectors. Plasmid sequence fidelity was verified by Sanger sequencing (Johns Hopkins Genomic Research Core Facilities). Plasmids were packaged into BaeV-pseudotyped replicationincompetent retroviral particles produced in HEK-293T cells using standard procedure.15[000249] CCR-NK cell production. NK cell activation, expansion, and engineering is performed as per lab standard.15Briefly, peripheral blood mononuclear cells from healthy donor leucopaks will be isolated using density gradient centrifugation. T cells are depleted using CD3-microbeads (Militenyi Biotec). CD3-depleted cells are plated and stimulated on Day 0 with K562.mbIL15.41BBL feeder cells.22Cells are maintained in Advanced RPMI media with 10% FBS, GlutaMAX, and 50 lU / mL recombinant human IL-2. Purity of NK cells are verified using flow cytometry and antibody-conjugated fluorophores. NK cells are transduced on Day 4 in 24- well plates containing RetroNectin-immobilized viral vector. CCR expression is validated using flow cytometry with his-tagged rhCD123 and fluorophore conjugated anti-his antibodies.[000250J CCR-NK cell systemic toxicity and anti-AML activity. We have shown that constitutive IL- 15 secretion by adoptively transferred NK cells is associated with lethal toxicity in vivo}~ Sustained proliferation of CCR-yc + CCR-IL-15RP NK cells in the presence of antigen as well as the ability of the NK cell products to clear leukemic blasts, without inducing inflammatory toxicity, will be tested and compared in our established human AML NSG xenograft models. All protocols are approved by the Johns Hopkins Institutional Animal Care and Use Committee.[000251] Briefly, 6 to 8-week-old female NSG (SlOV).Cg-PrkdcsctdIl2r^mIWjlISz2') mice will be injected via their tail veins with AML cells expressing ffLuc (i.e., Molm-13.ffLuc) on day 0. CCR-yc + CCR-IL-15RP NK cells will be injected after 7 days. Target cell bioluminescence will be tracked and quantified following intraperitoneal injection of D-luciferin using an IVIS Spectrum Imaging System each week. Blood will be collected weekly by submandibular bleeding. Peripheral blood will be used to quantify the number of circulating human NK cells per pL of blood by flow cytometry and NK-specific antibodies (e g., CD56, CD16, NKp30). Symptoms of toxicity will be monitored by weekly weight measurements and visual inspection of animals. If it is suspected that animals are afflicted with inflammatory disease, proinflammatory murine cytokines and cytokines produced by human NK cells will be determined by performing ELISA on plasma separated from collected blood.[000252] Statistical plan. All in vitro experiments are performed using at least three independent cell donors. Statistical significance is determined using multifactor ANOVA with Bonferroni correction and p-values < 0.05. Power analysis was performed to determine the number of mice needed for in vivo experiments. Mice will be treated with CCR-NK cells derived from at least two unique donors to account for donor-to-donor variability.[000253] Example 4:[000254] CCRs induce antigen-specific signaling and activation in response to stimulation with CD123 and includes a series of graphs and a blot demonstrating that CCRs induce antigenspecific signaling and activation in response to stimulation with CD 123. FIG. 4A: Nontransduced, 15Dl.mSc, ycDl.GFP, and co-transduced NK cells were stimulated with plate-bound rhCD123 or rhIL-2 (lane 3, positive control for antibody) for 30 minutes. Unstimulated cells served as negative controls. Cells were lysed, total protein quantified by BCA, and then western blot was performed on 20 pg of total protein from each lysate. STAT5 is phosphorylated (pSTAT5) downstream of yc cytokine family receptors. Antibody stains were used for total STAT5 and pSTAT5 with a GAPDH loading control. Lanes were quantified in imaged, normalized to GAPDH, and the ratio of pSTAT5:total STAT5 was calculated. Co-transduced NK cells showed a 2-fold increase in signal when stimulated with rhCD123. N = 3 NK cell donors.[000255] FIG. 4B: To assess if functional signaling was necessary for CCR functionality in NK cells, mutated ycDl.GFP constructs were created using site-directed mutagenesis. Two signaling null mutants were created by introducing an early stop codon after the transmembrane region (R289*, mutation of residue corresponding to R289 in the native yc) or by deleting the JAK binding motif (Boxl A, deletion of residues corresponding to T286 through K294 in the native yc). All CCR-yc constructs showed similar levels of transduction in NK cells as detected by flow cytometry using his-tagged rhCD123 and anti-his-tag antibody staining. To assess if functional scFv-binding was necessary for CCR functionality in NK cells, a native yc fused to GFP (yc-GFP) was cloned into an pSFG expression vector. Expression of yc-GFP was flow plots. Cells cotransduced with a CCR-pair showed high antigen-specific killing efficacy when co-cultured with MOLM-13 (1 :2 p <0.0001, 1: 1 p <0.0001 vs. non-transduced). NK cells did not show different cytotoxicity against Raji (all p > 0.6691 vs. non-transduced).[000256][000257] FIG. 4C: NK cells were transduced with mutated chimeric ycDl .GFPs (R289* or Boxl A), yc-GFP, and chimeric ycDl.GFP alone and in combination with our best expressing CCR- IL- 15R0 (15D 1.mSc). Engineered NK cells were co-cultured at a 1 : 1 effector-to-target ratio (E:T,where NK cells are effector cells and target cells are cancer cell lines) with firefly luciferase (ffLuc) expressing MOLM-13 for 18 hours. Reduction in bioluminescent signal was detected and compared to signal from target cells incubated without NK cells. Only NK cells co-transduced with CCR-IL-15RP and CCR-ycDl (ycDl.GFP with intact signaling and antigen-binding) were able to activate NK cells vs. MOLM-13 and induce cytotoxicity. N = 1, 3 technical replicates per condition. FIG. 4D: NK cells engineered with functional ycDl.GFP and 15Dl.mSc, alone or together, were co-cultured with ffLuc target cells, either MOLM-13 (CD 123 -positive) or Raji (CD 123 -negative), for 18 hours at various E:T ratios. Cytotoxicity was calculated as a reduction in bioluminescent signal compared to signal from cancer cells incubated without NK cells. Only NK cells co-transduced with both halves of our CCR-pair displayed elevated cytotoxicity. Statistical comparison was completed using two-way ANOVA with Tukey tests for multiple comparisons: **** p < 0.0001 vs. non-transduced, N = 3 NK cell donors, 3 technical replicates per donor.[000258] Example 5: High expression of both CCR chains induces robust cytotoxic activity against cancer cell lines.[000259] NK cells were engineered with monocistronic CCR-yc (ycDl), monocistronic CCR-IL-15RP (15D1), or bicistronic CCRs (YCD1.T2A.15D1) encoded in pMSGV expression vectors. CCR chains expressed individually or together displayed long-term surface expression and antigen binding up to 22 days post-activation, as determined by flow cytometry using his- tagged rhCD123 and anti-his-tag antibody staining. N = 3 NK cell donors. See FIG. 5A.[000260] CRISPR was used to knockout (KO) the target antigen of interest (CD 123) from three AML cell lines (MOLM-13, MV-4-11, and OCLAML3) to create negative controls. KO was confirmed using flow cytometry as shown in FIG 5B.[000261] NK cells engineered with monocistronic CCR chains or bicistronic CCRs were cocultured with ffLuc-expressing target cells, either CD 123 -positive targets (top: MOLM-13, MV411, 0CI-AML3, Raji engineered with constitutive CD123 expression) or CD 123 -negative targets (bottom: CRISPR knockout cell lines or parental Raji), for 18 hours at various E:T ratios. Reduction in bioluminescent signal was detected and compared to signal from target cellsincubated without NK cells. NK cells expressing both halves of our CCR-pair on one expression vector showed elevated cytotoxicity against all evaluated cancer cell lines shown in FIG. 5C. Statistical comparison was completed using two-way ANOVA with Tukey tests for multiple comparisons: * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001, color of asterisks corresponds to experimental condition vs. non-transduced, N = 3 NK cell donors, 3 technical replicates per donor. Higher activity versus all the cancer cell lines may be due to higher expression of the bicistronic CCRs (and higher tonic signaling): ycDl-GFP + 15Dl-mSc cotransduced - Day 8 GFP+mSc+His+: 36.3 ± 9.4% (FIG. 3B) Bicistronic - Day 8 His+: 54.5 ± 3.5% (FIG. 5 A).[000262] Example 6 CCR-NK cells have increased survival and sustained killing activity in vitro.[000263] NK cells were transduced with a BaeV-pseudotyped vector encoding a nuclear- localized (NLS) mScarlet alone (mSc) or with a 1 : 1 mixture of NLS-mSc vector and our bicistronic CCR vector (ycDl.T2A.15D1, CCR+mSc). Expression of mSc and CCRs were confirmed using flow cytometry and recombinant protein staining as shown in FIG 6A.[000264] NLS-mScarlet enabled long-term tracking of NK cells in an Incucyte S3 Live-Cell Analysis System. Engineered NK cells expressing CCRs or not were tracked over the course of 10 days with or without supplemental rhIL-2 (25 lU / mL, top). Media was refreshed every 2 days. CCR-NK cells displayed improved cytokine-independent survival but were still responsive to exogenous stimulation with cytokine. NK-cell numbers were quantified as the number of red objects normalized to the starting value at time 0 (FIG. 6B) where solid bold lines represent mean NK cell counts, while thin and dashed lines are individual replicates. NK cells were also serially stimulated with NLS-GFP expressing MOLM-13 and MOLM-13.CD123KO target cells (bottom). NK cells and AML targets were plated at an initial density of 2e5:le5 and le5 additional target cells were added every 2 days. CCR-NK cells were able to kill AML across consecutive stimulations. CD123+ parental MOLM-13 had a shorter half-live than MOLM-13.CD123KO cocultured with CCR-NK cells for all stimulations except the last. Quantity of AML was calculated as the number of green objects and normalized to the number of AML cells present at the start of each repeat stimulation. Cell numbers were quantified using the Incucyte 2022B Rev2 software.AML count curve comparisons were done by fitting one-phase decay curves to each stimulation using GraphPad Prism 10. Previously collected data using chimeric antigen receptor (CAR) modified NK cells has been included for comparison. N = 3 NK cell donors, 3 technical replicates per donor, 4 images per replicate. FIG. 6C: Representative images and respective image masks (counted objects) of engineered NK cells cultured with and without rhIL-2 on day 5 of 10-day culture. FIG. 6D: Following 10-day culture with and without rhIL-2, NK cells were harvested for flow cytometry. CCR+ cells, as determined by recombinant antigen staining (His+), had a trend of enrichment without cytokine supplementation but were not significantly elevated. Statistical comparison was completed using one-way ANOVA with Tukey tests for multiple comparisons. N = 3 NK cell donors[000265] Example 7: CCR-NK cells have increased survival in vivo.[000266] The longevity of CCR-NK cells was evaluated in 8-week-old female NSGmice. Mice were first engrafted with le6 MV-4-11 via tail vein injection. Mice were treated with saline, NK cells without a chimeric construct, or CCR-NK cells 7 days following AML injections with N = 4 or 5 mice per group. NK cells were modified to express firefly luciferase (ffLuc) to enable tracking in live animals. CCR+ffLuc.NK cells were transduced with separate viral vectors on subsequent days. See FIG. 7A.[000267] NK cell abundance was detected by bioluminescence imaging using an Xenogen IVIS Spectrum in vivo imaging system following intraperitoneal injection of D-luciferin. See FIG. 7B. Radiance was quantified using the Living Image software. In FIG. 7B, Thick lines represent means while thin dashed lines represent radiance signal for individual animals. FIG. 7C: Images collected for mice treated with ffLuc.NK and CCR+ffLuc.NK cells. FIG. 7D: One month post-treatment, all mice were sacrificed. Spleens, bone marrow (BM) from hind limbs, and peripheral blood (PB) was collected and processed for analysis by flow cytometry. NK cell abundance (top) and AML burden (bottom) was determined for each compartment. In FIG. 7D, in a specific pit, saline is plotted on the left, ffLuc.NK plotted in the central or middle position, and CCR+ffLuc.NK is plotted on the right.[000268] Example 8: CCRs can be functionalized against another cancer-associated antigen: B7H3.[000269] FIG. 8A: NK cells were engineered with monocistronic anti-B7H3 CCR-yc, monocistronic anti-B7H3 CCR-IL-15R0, or bicistronic CCRs encoded in pMSGV expression vectors. CCR chains could be expressed individually or together, as determined by flow cytometry on day 8 post NK-cell activation using his-tagged rhB7H3 and anti-his-tag antibody staining. N = 2 NK cell donors. FIG. 8B: CR1SPR was used to knockout (KO) the second target antigen of interest (B7H3) from two AML cell lines (MV-4-11 and OCI-AML3) to create negative controls. KO was confirmed using flow cytometry. FIG. 8C: NK cells engineered with anti-B7H3 CCR chains were co-cultured with either B7H3-positive or B7H3-negative ffLuc-expressing target cells for 18 hours at various E:T ratios. Reduction in bioluminescent signal was detected and compared to signal from target cells incubated without NK cells. CCR-NK cells expressing both chimeric anti-B7H3 chains showed elevated cytotoxicity against the evaluated cancer cell lines. Statistical comparison was completed using two-way ANOVA with Tukey tests for multiple comparisons: * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001, color of asterisks corresponds to experimental condition vs. non-transduced, N = 2 NK cell donors, 3 technical replicates per donor.

Claims

What is claimed:

1. A chimeric cytokine receptor comprising: i. a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor domains, an interleukin receptor transmembrane domain, and an interleukin receptor intracellular domain; ii. a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor common gamma chain (yc) domains, a yc transmembrane domain, and a yc intracellular domain.

2. The chimeric cytokine receptor of claim 1, wherein the one or more extracellular interleukin receptor domains comprise one or more interleukin 15 receptor (IL-15R) extracellular domains or variants thereof.

3. The chimeric cytokine receptor of claim 1, wherein the one or more transmembrane interleukin receptor domains comprise one or more interleukin 15 receptor (IL-15R) transmembrane domains or variants thereof.

4. The chimeric cytokine receptor of claim 1, wherein the one or more intracellular interleukin receptor domains comprise one or more interleukin 15 receptor (IL-15R) intracellular domains or variants thereof.

5. The chimeric cytokine receptor of claims 1 or 2, wherein the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 50% shorter than a wild type IL-15R extracellular domain.

6. The chimeric cytokine receptor of claims 1 or 2, wherein the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 75% shorter than a wild type IL-15R extracellular domain.

7. The chimeric cytokine receptor of claims 1 or 2, wherein the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 90% shorter than a wild type IL-15R extracellular domain.

8. The chimeric cytokine receptor of claims 1 or 2, wherein the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 95% shorter than a wild type IL-15R extracellular domain.

9. The chimeric cytokine receptor of claims 1 or 2, wherein the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is at least 99% shorter than a wild type IL-15R extracellular domain.

10. The chimeric cytokine receptor of claims 1 or 2, wherein the one or more extracellular domains of the IL-15R comprise an amino acid sequence length that is as long as a wild type IL- 15 extracellular domain.

11. The chimeric cytokine receptor of claim 1, wherein the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 50% shorter than a wild type yc extracellular domain.

12. The chimeric cytokine receptor of claim 1, wherein the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 75% shorter than a wild type yc extracellular domain.

13. The chimeric cytokine receptor of claim 1, wherein the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 90% shorter than a wild type yc extracellular domain.

14. The chimeric cytokine receptor of claim 1, wherein the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 95% shorter than a wild type yc extracellular domain.

15. The chimeric cytokine receptor of claim 1, wherein the one or more extracellular domains of the yc comprise an amino acid sequence length that is at least 99% shorter than a wild type yc extracellular domain.

16. The chimeric cytokine receptor of claim 1, wherein the one or more extracellular domains of the yc comprise an amino acid sequence length that is as long as a wild type yc extracellular domain.

17. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises an amino acid sequence identity of at least 75% to SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

18. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises an amino acid sequence identity of at least 80% to SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

19. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises an amino acid sequence identity of at least 90% to SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

20. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises an amino acid sequence identity of at least 95% to SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

21. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

22. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises a nucleic acid sequence identity of at least 75% to SEQ ID NOs: 1 or 3.

23. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises a nucleic acid sequence identity of at least 80% to SEQ ID NOs: 1 or 3.

24. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises a nucleic acid sequence identity of at least 90% to SEQ ID NOs: 1 or 3.

25. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises a nucleic acid sequence identity of at least 95% to SEQ ID NOs: 1 or 3.

26. The chimeric cytokine receptor of any one of claims 1-16, wherein the cytokine receptor comprises SEQ ID NOs: 1 or 3.

27. The chimeric cytokine receptor of any one of claims 1-26, wherein the antigen specific binding domain specifically binds to tumor antigens or virus antigens.

28. The chimeric cytokine receptor of any one of claims 1-26, wherein the antigen specific binding domain specifically binds cluster of differentiation antigen 123 (CD123) or 276 (CD276, B7H3).

29. The chimeric cytokine receptor of any one of claims 1-28, wherein the antigen specific binding domain comprises an antibody, an antibody binding fragment, an aptamer, or a peptide.

30. The chimeric cytokine receptor of any one of claims 1-28, wherein the antigen specific binding domain comprises a single chain variable fragment (scFv).

31. A vector comprising a cytokine receptor nucleic acid sequence having a nucleic acid sequence identity of at least 75% to SEQ ID NOs: 1 or 3.

32. The vector of claim 31, wherein the vector comprises a cytokine receptor nucleic acid sequence having a nucleic acid sequence identity of at least 95% to SEQ ID NOs: 1 or 3.

33. The vector of claim 31, wherein the vector comprises a cytokine receptor nucleic acid sequence comprising SEQ ID NOs: 1 or 3.

34. A vector comprising a nucleic acid sequence encoding a cytokine receptor amino acid sequence having an amino acid sequence identity of at least 75% to SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

35. The vector of claim 34, wherein the vector comprises a nucleic acid sequence encoding a cytokine receptor amino acid sequence having an amino acid sequence identity of at least 95% to SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

36. The vector of claim 34, wherein the vector comprises a nucleic acid sequence encoding a cytokine receptor amino acid sequence comprising SEQ ID NOs: 2, 4, 4A, 5, 5A, 6 or 6A.

37. A pharmaceutical composition comprising a chimeric cytokine receptor of any one of claims 1-30 or a vector of any one of claims 31-36.

38. A host cell comprising a vector of any one of claims 31-36.

39. The host cell of claim 38, wherein the host cell comprises an autologous cell, an allogeneic cell, a haplotype matched cell, a haplotype mismatched cell, a haplo-identical cell, a xenogeneic cell, a stem cell, cell lines or combinations thereof.

40. The host cell of claim 39, wherein the host cell comprises an immune cell.

41. The host cell of claim 40, wherein the immune cell is a natural killer (NK) cell.

42. A method of modulating natural killer (NK) cell persistence in vivo, comprising: administering to a subject, a pharmaceutical composition comprising a therapeutically effective amount of a chimeric cytokine receptor, or an NK cell transduced with a vector encoding for the chimeric cytokine receptor, wherein the chimeric cytokine receptor comprises: i. a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin 15 receptor (IL-15R) domains, an IL-15R transmembrane domain, and an IL-15R intracellular domain; ii. a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin common gamma chain (yc) domains, a yc transmembrane domain, and a yc intracellular domain.

43. The method of claim 38, wherein the NK cells comprise an autologous cell, an allogeneic cell, a haplotype matched cell, a haplotype mismatched cell, a haplo-identical cell, or combinations thereof.

44. The method of claim 43, wherein the NK cell is an autologous cell.

45. The method of claim 44, wherein the NK cells are transduced ex vivo with the chimeric cytokine receptor molecule.

46. The method of claim 45, wherein the transduced NK cells are adoptively transferred to the subject.

47. The method of any one of claims 42-46, further comprising administering one or more secondary therapies.

48. The method of claim 47, wherein the one or more secondary therapies comprise chemotherapeutic agents, immune therapeutic agents, checkpoint inhibitors, radiation, surgery or combinations thereof.

49. A method of treating a virus infection or cancer, comprising administering to a subject, a pharmaceutical composition comprising a therapeutically effective amount of a chimeric cytokine receptor, or an isolated cell transduced with a vector encoding for the chimeric cytokine receptor, wherein the chimeric cytokine receptor comprises: i. a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor domains, an interleukin receptor transmembrane domain, and an interleukin receptor intracellular domain; ii. a single chain variable fragment (scFv) which specifically binds to an antigen, one or more extracellular interleukin receptor common gamma chain (yc) domains, a yc transmembrane domain, and a yc intracellular domain.

50. The method of claim 49, wherein the isolated cell comprises an autologous cell, an allogeneic cell, a haplotype matched cell, a haplotype mismatched cell, a haplo-identical cell, or combinations thereof.

51. The method of claim 50, wherein the isolated cell is an autologous cell.

52. The method of claim 50, wherein the isolated cells are transduced ex vivo with the chimeric cytokine receptor molecule.

53. The method of claim 50, wherein the transduced isolated cells are adoptively transferred to the subject.

54. The method of any one of claims 49-53, further comprising administering one or more secondary therapies.

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