MANUFACTURING METHODS FOR ALLOGENIC CAR-T CELLS.

Abstract

This description outlines improved media for culturing immune cells and methods for using them. In particular, the cell culture media described herein are especially suitable for the expansion of T cells, which can be used to manufacture cells useful in adoptive cell therapies, including therapies using chimeric antigen receptors (e.g., CAR-T cell therapy).

Description

MANUFACTURING METHODS FOR ALLOGENIC CAR-T CELLS CROSS REFERENCE This application claims the benefit of priority to United States provisional application no. 62 / 839,449, filed on April 26, 2019, the contents of which are hereby incorporated by reference in their entirety. TECHNICAL FIELD This description relates to cell culture media and methods for the manufacture of manipulated immune cells, including those comprising chimeric antigen receptors (CARs) and manipulated T cell receptors (TCRs), and methods for treating cancer in a patient by using the same. LIST OF SEQUENCES This request contains a Sequence List that was submitted electronically in ASCII format and is therefore incorporated herein by reference in its entirety. This ASCII copy, created on April 17, 2020, is named AT-025_02WO_ST25.txt and is 15,405 bytes in size. BACKGROUND Adoptive transfer of immune cells genetically modified to recognize malignancy-associated antigens shows promise as a novel approach to cancer treatment (see, for example, Brenner et al., Current Opinion in Immunology, 22(2): 251-257 (2010); Rosenberg et al., Nature Reviews Cancer, 8(4): 299-308 (2008)). Immune cells can be genetically modified to express chimeric antigen receptors (CARs) (see, for example, Eshhar et al., Proc. Nati. Acad. Sci. USA, 90(2): 720-724 (1993) and Sadelain et al., Curr. Opin. Immunol., 21(2): 215-223 (2009)). CAR-containing immune cells, for example, CAR-T cells (CAR-T), are manipulated to give them antigenic specificity while retaining or improving their ability to recognize and destroy a target cell.Effective immune cell culture media and methods for using them can be particularly useful for manufacturing engineered immune cells such as CAR-T cells. This description provides cell culture media and manufacturing methods that address this need. SUMMARY This description outlines improved media for culturing immune cells and methods of use. For example, this description includes media that are particularly suitable for T-cell expansion, which can be used to manufacture cells useful in adoptive cell therapies, including therapies using chimeric antigen receptors (e.g., CAR-T cell therapy). In one aspect, the description provides a cell culture medium for T cell expansion comprising: a first cell proliferation stimulant and a second cell proliferation stimulant, each independently selected from the group consisting of IL-4, IL-7, IL-10, IL-12, and IL-15, and wherein the first stimulant and the second stimulant are present in a concentration ratio of approximately 1,000:1 to approximately 4:1. In one aspect, the description provides a cell culture medium for T cell expansion comprising an extracellular modulator of cell metabolism which is extracellular potassium at a concentration of approximately 4 mM to approximately 40 mM. In one aspect, the description provides a cell culture medium for T cell expansion comprising: a first and a second cell proliferation stimulant, each independently selected from the group consisting of IL-4, IL-7, IL-10, IL-12, and IL-15; and an extracellular modulator of cell metabolism, which is extracellular potassium at a concentration of approximately 4 mM to approximately 40 mM; and wherein the first and second stimulants are present in a concentration ratio of approximately 1,000:1 to approximately 4:1. In some modalities, the first stimulant of cell proliferation is IL 7. In some modalities, the second stimulant of cell proliferation is IL 15. In some forms, the first stimulant and the second stimulant are present in a concentration ratio of approximately 500:1 to approximately 10:1, approximately 250:1 to approximately 10:1, approximately 200:1 to approximately 10:1, approximately 150:1 to approximately 10:1, approximately 100:1 to approximately 10:1, approximately 500:1 to approximately 50:1, approximately 250:1 to approximately 50:1, approximately 200:1 to approximately 50:1, approximately 150:1 to approximately 50:1, approximately 100:1 to approximately 50:1, approximately 500:1 to approximately 75:1, approximately 250:1 to approximately 75:1, approximately 200:1 to approximately 75:1, approximately 150:1 to approximately 75:1, approximately 100:1 to approximately 75:1, approximately 10:1 to approximately 4:1, approximately 8:1 to approximately 4:1 or approximately 7:1 to approximately 6:1. In some modalities, the first stimulant and the second stimulant are present in a concentration ratio of approximately 150:1, approximately 140:1, approximately 130:1, approximately 120:1, approximately 110:1, approximately 100:1, approximately 90:1, approximately 80:1, or approximately 70:1. In some modalities, a first stimulant is IL-7, present at a concentration of approximately 100 Ul / ml to approximately 5,000 Ul / ml; and a second stimulant is IL15, present at a concentration of approximately 1 Ul / ml to approximately 100 Ul / ml. MA / a / ¿U¿l / Ul dUUO In some modalities, IL-7 is present at a concentration of approximately 300 Ul / ml to approximately 5,000 Ul / ml and IL-15 is present at a concentration that is approximately 25 Ul / ml to approximately 50 Ul / ml. In some modalities, IL-7 is present at a concentration of approximately 5,000 Ul / ml and IL-15 is present at a concentration that is approximately 50 Ul / ml. In some modalities, the first stimulant and the second stimulant are added to the cell culture simultaneously or sequentially. In some modalities, extracellular potassium is present at a concentration of approximately 4 mM to approximately 40 mM; approximately 10 mM to approximately 35 mM; approximately 10 mM to approximately 25 mM; approximately 20 mM to approximately 35 mM; approximately 20 mM to approximately 25 mM; or approximately 20 mM to approximately 30 mM. In some modalities, extracellular potassium is present at a concentration of approximately 20 mM or approximately 25 mM. In some forms, extracellular potassium is present at a concentration less than 40 mM and greater than 4 mM; less than 40 mM and greater than 10 mM; less than 40 mM and greater than 20 mM; or less than 40 mM and equal to or greater than 25 mM. In some forms, extracellular potassium is present as KCl. In some modalities, the T cell population obtained is enriched in Tcm and / or TsCM cells. In another aspect, the description provides a method for obtaining a T cell population in vitro, comprising culturing an initial T cell population with a cell culture medium comprising a first and a second cell proliferation stimulant, each independently selected from the group consisting of IL-4, IL-7, IL-10, IL-12, and IL-15; and an extracellular modulator of cell metabolism, which is extracellular potassium at a concentration of approximately 4 mM to approximately 40 mM; and wherein the first and second stimulants are present in a concentration ratio of approximately 1,000:1 to approximately 4:1, and wherein the resulting T cell population is enriched in Tcm and / or Tscm cells. In some modalities, the cell culture medium is the cell culture medium as described above. In some modalities, the T cell population obtained comprises at least approximately 30%, 35%, 40%, or 45% Tscm cells. In some modalities, the T cell population obtained comprises at least approximately 40% Tscm cells. MA / a / ZUZl / Ul dUUO In some modalities, the T cell population obtained is approximately 100 to approximately 1,000 times that of the initial T cell population measured over a period of approximately 7-16 days. In some modalities, the T cell population obtained is approximately 100 to approximately 1,000 times that of the initial T cell population measured over a period of approximately 10-14 days. In some modalities, the T cell population obtained is at least approximately 100 or approximately 200 times that of the initial T cell population. In some modalities, the resulting T cell population is enriched approximately 100 to approximately 1,000 times in Tcm and / or Tscm than the initial T cell population, measured over a period of approximately 7–16 days. In some modalities, the resulting T cell population is enriched approximately 100 to approximately 1,000 times in Tcm and / or Tscm than the initial T cell population, measured over a period of approximately 10–14 days. In some modalities, the resulting T cell population is enriched at least approximately 100 or at least approximately 200 times in Tcm and / or Tscm than the initial T cell population. In some modalities, the initial T cell population is a manipulated T cell population. In some modalities, the initial T cell population is a population of T cells that express one or more chimeric antigen receptors. In some modalities, the T cells are allogeneic T cells. In some modalities, the T cells are autologous T cells. In one aspect, the description provides a population of manipulated immune cells, wherein said population comprises T cells expressing one or more chimeric antigen receptors (CAR-T cells), and wherein said CAR-T cells are obtained by using the cell culture medium as described above. In one aspect, the description provides a population of manipulated immune cells, wherein said population comprises T cells expressing one or more chimeric antigen receptors (CAR-T cells), and wherein said (CAR-T cells) are obtained by using the method as described above. In some modalities, CAR-T cells comprise at least approximately 30%, 35%, 40%, or 45% Tscm cells. In some modalities, CAR-T cells comprise at least approximately 40% Tscm cells. In one respect, the description provides a pharmaceutical composition comprising the manipulated immune cell population as described above. In one aspect, the description provides a method for treating a disease or disorder in a subject in need, comprising administering to the subject the manipulated immune cell as described above or the pharmaceutical composition as described above. In some cases, the disease or disorder is cancer. In one respect, the description provides a manufactured article comprising the manipulated immune cell as described above or the pharmaceutical composition as described above. All the modalities presented in this description are applicable to all aspects described throughout the description. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1A represents a comparison of CAR T+ phenotypes using processes comprising: sequential stimulation with IL-2; stimulation with IL-2 followed by stimulation with IL-7 + IL-15; and sequential stimulation with IL-7 + IL-15. The IL-7 + IL-15-based processes increase the abundance of Tscm CAR-T cells. Figure 1B represents the cytokine release capabilities of CD19-specific CAR-T cells after exposure to target cells, where the CAR-T cells were prepared using manufacturing processes comprising sequential stimulation with IL-2; stimulation with IL-2 followed by stimulation with IL-7 + IL-15; and sequential stimulation with IL-7 + IL-15. Figure 2A represents the effect of increased extracellular potassium (40 mM, unfilled bars) on increasing Tscm cell abundance in IL-2 and IL-7+15 processes compared to normal potassium concentrations (4 mM) (filled bars). Figure 2B represents the effect of increased extracellular potassium (40 mM, unfilled bars) on the expansion capacity of allogeneic CAR-T cells in both IL-2 and IL-7+15 processes compared to normal potassium concentrations (4 mM, filled bars). Figure 3 depicts the effect of extracellular potassium concentrations of 25 mM and 10 mM: these concentrations did not negatively affect the expansion of human T cells during the fabrication of allogeneic CAR-T cells compared to higher concentrations. Figure 4 shows that the maximum preservation of Tscm cells was achieved at extracellular potassium 25 mM. Figure 5 shows that enhanced potency of Flt3-specific CAR-T cells (Allo-819) was obtained by using a combination of an IL-7+15-based process with 25 mM extracellular potassium supplementation. DETAILED DESCRIPTION This description outlines cell culture media and methods particularly useful for the cultivation of immune cells. Adoptive cell therapy using engineered immune cells is a type of immunotherapy that has emerged as a particularly promising new approach to cancer treatment. The production and manufacture of engineered immune cells involves collecting immune cells (e.g., T cells) from a subject followed by in vitro cell expansion. Successful in vitro expansion of immune cells (e.g., T cells) is characterized by sufficient cell proliferation as well as the development of desirable cell phenotypes. For example, T cell expansion can result in a mixture of T cells that includes subsets of naïve T cells (TN), memory T cells (including stem cell-like memory T cells (Tscm), core memory T cells (Tcm), and effector memory T cells (Tem)), and effector T cells (Tefe). The varying amounts of different T cell subsets can affect the therapeutic profile and efficacy of the resulting manipulated T cells. In particular, T cell expansion methods that yield enriched quantities of T cells in early stages of differentiation are therapeutically advantageous. Consequently, the cell culture media described herein may be particularly beneficial for T cell expansion where the ratio of Tscm and / or Tcm cells is enriched. Tscm cells are the least differentiated type of memory T cell and, for adoptive T cell therapy, may be particularly advantageous for promoting T cell proliferation in vivo after administration of the manipulated cells to a patient. Therefore, such cell culture media and immune cells (e.g., T cells) prepared using such media may result in more potent adoptive cell transfer therapies, including CAR-T cell therapies as described herein. In particular, cell culture media comprising combinations of different cell proliferation stimulants (e.g., a first and second T cell growth factor) in combination with an extracellular modulator of cell metabolism can result in convenient cell proliferation and cell phenotypes. MA / a / ZUZl / Ul dUUO In one respect, the description presents a cell culture medium (e.g., a medium suitable for T cell expansion) comprising: a first stimulant of cell proliferation (e.g., a first cytokine that stimulates T cell proliferation); a second cell proliferation stimulant (e.g., a second cytokine that stimulates T cell proliferation); and an extracellular modulator of cell metabolism (e.g., T cell metabolism). In particular, the combinations and concentrations of cytokines and metabolic modulators10 as described herein provide novel means of T-cell expansion that may lead to increased potency of immune cell therapy products, including allogeneically modified cell therapy products and autologous cell therapy products. In one modality, the combination of the stimulators IL-7 and IL-15 along with increased extracellular potassium may achieve15 improvements in obtaining desirable phenotypes of allogeneic CAR-T cells (e.g., increased Tscm cell population) and / or autologous cell therapy products compared to classical IL-2-based expansion conditions. 1. Immune cells Cells suitable for cultivation using the media and methods described herein include immune cells. Prior to in vitro genetic manipulation or modification (e.g., as described herein), cells for use in the methods described herein (e.g., immune cells) may be obtained from a subject. Cells25 may be obtained from various non-limiting sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, stem cell-derived immune cells or iPSCs, tissue from an infection site, ascites, pleural effusion, spleen tissue, and tumors. In some modalities, any variety of T cell lines available and known to those skilled in the technique may be used.30 In some modalities, cells may be derived from a healthy donor, a patient diagnosed with cancer, or a patient diagnosed with an infection.In some modalities, the cells may be part of a mixed population of cells that exhibit different phenotypic characteristics. In some methods, immune cells are obtained from a subject who will ultimately receive the engineered immune cells. In other methods, immune cells are obtained from a donor, who is a different individual from the subject who will receive the engineered immune cells. In some modalities, immune cells comprise T cells. T cells can be obtained from various sources, including peripheral blood mononuclear cells (PBMCs), bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, stem cell-derived T cells or iPSCs, tissue from an infection site, ascites, pleural effusion, spleen tissue, and tumors. In certain modalities, T cells can be obtained from a blood volume collected from the subject using any variety of techniques known to the practitioner, such as FICOLL™ separation. Cells can be obtained from an individual's circulating blood by apheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets. In certain modalities, the cells collected by apheresis can be washed to remove the plasma fraction and placed in an appropriate buffer or medium for subsequent processing. PBMCs can be used directly for genetic modification of immune cells (such as CAR or TCR cells) using methods as described herein. In certain modalities, after isolating the PBMCs, T lymphocytes can be further isolated, and cytotoxic and helper T lymphocytes can be sorted into naïve, memory, and effector T cell subpopulations before or after genetic modification and / or expansion. In certain modalities, T cells are isolated from PBMCs by lysis of red blood cells and depletion of monocytes, for example, by centrifugation through a PERCOLL™ gradient. A specific T cell subpopulation, such as CCR7+, CD95+, CD122+, CD27+, CD69+, CD127+, CD28+, CD3+, CD4+, CD8+, CD25+, CD62L+, CD45RA+, and CD45RO+ T lymphocytes, can be further isolated using positive or negative selection techniques known in the technique. For example, enrichment of a T cell population by negative selection can be achieved with a combination of antibodies targeting unique surface markers for negatively selected cells.One method for use in this description is cell sorting and / or selection by negative magnetic immunoadherence or flow cytometry using a cocktail of monoclonal antibodies targeting cell surface markers present on the negatively selected cells. For example, to enrich CD4+ cells by negative selection, a monoclonal antibody cocktail typically includes antibodies against CD14, CD20, CD11b, CD16, HLA-DR, and CD8. Flow cytometry and cell sorting can also be used to isolate cell populations of interest for use in this description. In some modalities, a population of T cells becomes enriched in C4+ cells. In some modalities, a population of T cells is enriched in CD8+ cells. In some modalities, CD8+ cells are further classified into naïve, central memory, and effector cells by identifying the cell surface antigens associated with each of these CD8+ cell types. In some modalities, the expression of phenotypic markers of central memory T cells includes CD45RO, CD62L, CCR7, CD28, CD3, and CD127, and they are negative for granzyme B. In some modalities, central memory T cells are CD45RO+, CD62L+, and / or CD8+ T cells. In some modalities, effector T cells are negative for CD62L, CCR7, CD28, and / or CD127, and positive for granzyme B and perforin. In certain modalities, CD4+ T cells are further classified into subpopulations. For example, CD4+ helper T cells can be classified into naïve, central memory, and effector cells by identifying cell populations that have cell surface antigens. It will be noted that PBMCs may also include other cytotoxic lymphocytes such as NK cells or NKT cells. An expression vector carrying the coding sequence of a chimeric receptor, as described herein, can be introduced into a population of human donor T cells, NK cells, or NKT cells. Successfully transduced T cells carrying the expression vector can be sorted using flow cytometry to isolate CD3-positive T cells and then further propagated to increase the number of these CAR-expressing T cells, in addition to cell activation using anti-CD3 and IL-2 antibodies or other methods known in the art, as described elsewhere herein. Standard procedures are used for the cryopreservation of CAR-expressing T cells for storage and / or preparation for use in a human subject.In one modality, the transduction, culture and / or in vitro expansion of T cells are performed in the absence of non-human animal-derived products such as fetal calf serum and fetal bovine serum. 302. Manipulated immune cells The cell culture media and methods of use described herein may be particularly useful in the in vitro expansion of immune cells, including manipulated immune cells (e.g., CAR-T cells). Manipulated immune cells can be allogeneic or autologous. In some modalities, the manipulated immune cell is a T cell (e.g., cytotoxic T lymphocyte, inflammatory T lymphocyte, regulatory T lymphocyte, helper T lymphocyte, tumor-infiltrating lymphocyte (TIL)), NK cell, NK-T cell, TCR-expressing cell, dendritic cell, killer dendritic cell, mast cell, or a B cell. In some modalities, the cell may be derived from the group consisting of CD4+ T lymphocytes and CD8+ T lymphocytes. In some illustrative modalities, the manipulated immune cell is a T cell. In some modalities, the manipulated immune cell can be derived from, for example, without limitation, a stem cell. Stem cells can be adult stem cells, non-human embryonic stem cells, more particularly non-human stem cells, umbilical cord blood stem cells, progenitor cells, bone marrow stem cells, induced pluripotent stem cells, totipotent stem cells, or hematopoietic stem cells. In some modality, the cell is obtained or prepared from peripheral blood. In some modality, the cell is obtained or prepared from peripheral blood mononuclear cells (PBMCs). In some modality, the cell is obtained or prepared from bone marrow. In some modality, the cell is obtained or prepared from umbilical cord blood. In some modality, the cell is a human cell. In some modality, the cell is transfected or transduced with the nucleic acid vector using a method selected from the group consisting of electroporation, sonoporation, biolistics (e.g., gene gun), lipid transfection, polymer transfection, nanoparticles, or polyplexes. a. Binding agents (including antibodies) In some embodiments, the manipulated immune cells comprise an antigen-binding agent (e.g., comprising an antigen-binding domain or comprising an antibody or fragment thereof). As used in this description, the term antibody refers to a polypeptide that includes sufficient canonical immunoglobulin sequence elements to confer specific binding to a particular target antigen. As known in the art, intact antibodies produced in nature are tetrameric agents of approximately 150 kDa composed of two identical heavy-chain polypeptides (approximately 50 kDa each) and two identical light-chain polypeptides (approximately 25 kDa each) that associate with each other in what is commonly known as a Y-shaped structure. Each heavy chain is composed of at least four domains (each approximately 110 amino acids in length): an amino-terminal variable domain (VH) (located at the tips of the Y structure), followed by three constant domains: CH1, CH2, and the carboxy-terminal CH3 (located at the base of the Y stem).A short region, known as the switch, connects the variable and constant regions of the heavy chain. The hinge connects the domains. CH2 and CH3 with the rest of the antibody. Two disulfide bonds in this hinge region connect the two heavy-chain polypeptides to each other in an intact antibody. Each light chain is composed of two domains—an amino-terminal variable domain (VL), followed by a carboxy-terminal constant domain (CL), separated from each other by another switch. Skilled in the art are familiar with antibody structure and sequence elements, recognize the variable and constant regions in the provided sequences, and understand that there may be some flexibility in defining a boundary between such domains, so that different presentations of the same antibody chain sequence may, for example, indicate such a boundary at a location that is offset by one or a few residues relative to a different presentation of the same antibody chain sequence. Intact antibody tetramers are composed of two heavy-chain-light-chain dimers in which the heavy and light chains are linked by a single disulfide bond; two additional disulfide bonds connect the hinge regions of the heavy chains, so that the dimers connect to each other and form the tetramer. Naturally produced antibodies are also glycosylated, typically in the CH2 domain. Each domain in a natural antibody has a structure characterized by an immunoglobulin fold formed from two beta sheets (e.g., 3-, 4-, or 5-chain sheets) packed together in a compressed antiparallel beta barrel. Each variable domain contains three hypervariable loops known as complementarity-determining regions (CDR1, CDR2, and CDR3) and four somewhat invariant structural framework regions (FR1, FR2, FR3, and FR4).When natural antibodies fold, the FR regions form beta sheets that provide the structural framework for the domains, and the CDR loop regions of the heavy and light chains approximate in three-dimensional space, creating a unique hypervariable antigen-binding site located at the tip of the Y-structure. The Fe region of natural antibodies binds to elements of the complement system and also to receptors on effector cells, including, for example, effector cells that mediate cytotoxicity. As is known in the art, the affinity and / or other binding attributes of Fe regions for Fe receptors can be modulated by glycosylation or other modification. In some embodiments, antibodies produced and / or used according to the present description include glycosylated Fe domains, which are Fe domains with such modified or manipulated glycosylation. For the purposes of this description, in certain embodiments, any polypeptide or polypeptide complex that includes sufficient immunoglobulin domain sequences as found in naturally occurring antibodies may be designated and / or used as an antibody, whether such polypeptide occurs naturally (e.g., generated by an organism reacting to an antigen) or is produced by recombinant engineering, chemical synthesis, or other artificial system or methodology. In some embodiments, an antibody is polyclonal; in some embodiments, an antibody is monoclonal. In some embodiments, an antibody has constant region sequences that are characteristic of mouse, rabbit, primate, or human antibodies. In some embodiments, the antibody sequence elements are humanized, primatized, chimeric, etc., as known in the art. Furthermore, the term antibody, as used herein, may refer in appropriate ways (unless otherwise indicated or evident from the context) to any of the constructs or formats developed or known in the art to utilize structural and functional features of the antibody in an alternative presentation. For example, in some embodiments, an antibody used according to herein is in a format selected from, but not limited to, intact IgA, IgG, IgE, or IgM antibodies; β- or multispecific antibodies (e.g., Zybodies®, etc.).); Antibody fragments such as Fab fragments, Fab' fragments, F(abj2) fragments, Fd' fragments, Fd fragments, and isolated CDRs or assemblies thereof; single-chain Fv; polypeptide-Fc fusions; single-domain antibodies (e.g., shark single-domain antibodies such as IgNAR or fragments thereof); camelid antibodies; masked antibodies (e.g., Probodies®); small modular immunopharmaceuticals (SMIPs™); single-chain or tandem diabodies (TandAb®); VHH; Anticalins®; Nanobodies® minibodies; B¡TE®s; ankyrin repeat proteins or DARPINs®; Avimers®; DART; TCR-like antibodies, Adnectins®; Affilins®; Trans-bodies®; Affibodies®; TrimerX®; MicroProteins; Fynomers®; Centyrins®; and KALBITOR®. In some forms, an antibody may lack a covalent modification. (for example, the joining of a glycan) that it would have if it occurred naturally.In some forms, an antibody may contain a covalent modification (e.g., attachment of a glycan, a payload (e.g., a detectable residue, a therapeutic residue, a catalytic residue, etc.) or another dangling group (e.g., polyethylene glycol, etc.). In some embodiments, the antibody or binding agent may be symmetric. Symmetric means that the antibody or binding agent has the same type of Fv regions (e.g., the antibody has two Fab regions). In some embodiments, the antibody or binding agent may be asymmetric. Asymmetric means that the antibody or binding agent has at least two different types of Fv regions (e.g., the antibody has Fab and scFv regions, Fab and scFv2 regions, or Fab-VHH regions). Several asymmetric architectures of antibodies or binding agents are known in the art (Brinkman and Kontermann et al. 2017 Mabs (9)(2): 182-212). As used herein, the term antibody agent refers to an agent that binds specifically to a particular antigen. In some modalities, the term includes any polypeptide or polypeptide complex that includes sufficient immunoglobulin structural elements to confer specific binding. Illustrative antibody agents include, but are not limited to, monoclonal or polyclonal antibodies. In some modalities, an antibody agent may include one or more constant region sequences that are characteristic of mouse, rabbit, primate, or human antibodies. In some modalities, an antibody agent may include one or more humanized, primatized, chimeric, etc., sequence elements, as known in the art.In many modalities, the term antibody agent is used to refer to one or more of the constructs or formats developed or known in the art to utilize the structural and functional characteristics of antibodies in an alternative presentation. For example, an antibody agent used according to the present description is in a format selected from, but not limited to, intact IgA, IgG, IgE, or IgM antibodies; β- or multispecific antibodies (e.g., Zybodies®, etc.).); antibody fragments such as Fab fragments, Fab' fragments, F(abj2) fragments, Fd' fragments, Fd fragments, and isolated CDRs or assemblies thereof; single-chain Fv; polypeptide-Fc fusions; single-domain antibodies (e.g., shark single-domain antibodies such as IgNAR or fragments thereof); camelid antibodies; masked antibodies (e.g., Probodies®); small modular immunopharmaceuticals (SMIPs™); single-chain or tandem diabodies (TandAb®); VHH; Anticalins®; Nanobodies® minibodies; BITE®; ankyrin repeat proteins or DARPINs®; Avimers®; DART; TCR-like antibodies; Adnectins®; Affilins®; Trans-bodies®; Affibodies®; TrimerX®; Microproteins; Fynomers®; Centyñns®; and KALBITOR®. An antibody or antigen-binding molecule encoded in this description may be single-stranded or double-stranded. In some embodiments, the antibody or antigen-binding molecule is single-stranded. In certain embodiments, the antigen-binding molecule is selected from the group consisting of scFv, Fab, Fab', Fv, F(abj2), dAb, and any combination thereof. Antibodies include antibody fragments. An antibody fragment comprises a portion of an intact antibody, such as the antigen-binding or variable region of the intact antibody. Antibody fragments include, but are not limited to, Fab, Fab', Fab'SH, F(abj2), Fv, diabody, linear antibodies, multispecific antibodies formed from antibody fragments, scFv antibodies and fragments, and other fragments described below. Antibodies also include, but are not limited to, monoclonal polyclonal antibodies, chimeric dAbs (domain antibodies), single-chain fragments, Fab, Fa, F <ab) 2, scFv y bibliotecas de expresión de Fab- Un anticuerpo puede ser un anticuerpo completo, una inmunoglobulina o un fragmento de anticuerpo.In some embodiments, the antibody is a full-length antibody, e.g., an intact lgG1 antibody or another class or isotype of antibody as described herein (see, e.g., Hudson et al., Nat. Med., 9:129-134 (2003); Pluckthun, The Pharmacology of Monoclonal Antibodies, vol. 113, pp. 269-315 (1994); Hollinger et al., Proc. Nati. Acad. Sci. USA, 90: 6444-6448 (1993); WO93 / 01161; and U.S. Patent Nos. 5,571,894, 5,869,046, 6,248,516 and 5,587,458). A full-length antibody, an intact antibody, or a complete antibody is an antibody that has a structure substantially similar to a native antibody structure or that has heavy chains containing an Fe region as defined in the present description.Antibody fragments can be prepared using various techniques, including, but not limited to, proteolytic digestion of an intact antibody, as well as production by recombinant host cells (e.g., E. coli or phage), as known in the technique. In some forms, an antibody is or comprises a monoclonal antibody, which includes a chimeric, humanized, or human antibody. In some embodiments, an antigen-specific antibody agent provided herein may be a chimeric antibody (see, for example, U.S. Patent No. 4,816,567; and Morrison et al., Proc. Nati. Acad. Sci. USA, 81: 6851-6855 (1984)). A chimeric antibody may be an antibody in which a portion of the heavy and / or light chain is derived from one particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species. In one example, a chimeric antibody may comprise a non-human variable region (for example, a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate such as a monkey) and a human constant region. In a further example, a chimeric antibody may be a class-switched antibody in which the class or subclass has been changed from that of the original antibody.Chimeric antibodies include antigen-binding fragments of the same. In some forms, a chimeric antibody can be a humanized antibody (see, for example, Almagro and Fransson, Front. Biosci., 13:1619-1633 (2008); Riechmann et al., Nature, 332:323-329 (1988); Queen et al., Proc. Nati Acad. Sci. USA 86:10029-10033 (1989); U.S. Patents Nos. 5,821,337, 7,527,791, 6,982,321 and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005); Padlan, Mol. Immunol., 28:489-498 (1991); Dall'Acqua et al., Methods, 36:43-60). (2005); Osbourn et al., Methods, 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000)). A humanized antibody is a chimeric antibody comprising amino acid residues from non-human hypervariable regions and amino acid residues from human FR.In certain configurations, 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 regions (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all of the receptors correspond to those of a human antibody. A humanized antibody may optionally comprise at least a portion of an antibody constant region derived from a human antibody. A non-human antibody can be humanized to reduce immunogenicity in humans while retaining the specificity and affinity of the original non-human antibody. A humanized antibody may comprise one or more variable domains comprising one or more CDRs, or portions thereof, derived from a non-human antibody. A humanized antibody may also comprise one or more variable domains comprising one or more FRs, or portions thereof, derived from human antibody sequences. Optionally, a humanized antibody may also comprise at least a portion of a human constant region. In some embodiments, one or more FR residues in a humanized antibody are replaced with the corresponding residues from a non-human antibody (e.g., the antibody from which the CDR residues are derived) to restore or enhance the antibody's specificity or affinity. Human scaffold regions that can be used for humanization include, but are not limited to: scaffold regions selected using a best-fit method; scaffold regions derived from the consensus sequence of human antibodies from a particular subset of light or heavy chain variable regions; mature (somatically mutated) human scaffold regions or human germline scaffold regions; and structural framework regions derived from screening FR libraries (see, for example, Sims et al., J. Immunol., 151:2296 (1993); Carter et al., Proc. Nati. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immunol., 151:2623 (1993); Baca et al., J. Biol. Chem., 272:10678-10684 (1997); and Rosok et al., J. Biol. Chem., 271:22611-22618 (1996)). In some embodiments, an antibody agent provided in this description is a human antibody. Human antibodies can be produced using various techniques known in the art (see, for example, van Dijk and van de Winkel, Curr. Opin. Pharmacol, 5: 368-74 (2001); and Lonberg, Curr. Opin. Immunol, 20:450-459 (2008)). A human antibody may be one that possesses an amino acid sequence corresponding to that of an antibody produced by a human being or a human cell, or derived from a non-human source that utilizes human antibody repertoires or other sequences encoding human antibodies. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues.Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigenic exposure (see, for example, Lonberg, Nat. Biotech., 23: 1117-1125 (2005); U.S. Patent Nos. 6,075,181, 6,150,584, 5,770,429, and 7,041,870; and U.S. Patent Application Publication No. US 2007 / 0061900). The human variable regions of intact antibodies generated by such animals can be further modified, for example, by combining them with a different human constant region. Human antibodies can also be prepared using hybridoma-based methods. For example, human antibodies can be produced from human myeloma and mouse-human heteromyeloma cell lines using human B-cell hybridoma technology and other methods (see, for example, Kozbor, J. Immunol, 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (1987); Boemer et al., J. Immunol, 147: 86 (1991); L1 et al., Proc. Nati. Acad. Sel. USA, 103: 3557-3562 (2006); U.S. Patent No. 7,189,826; Ni, Xiandai Mianyixue, 26(4): 265-268 (2006); Vollmers and Brandlein, Histology and Histopathology, 20(3): 927-937 (2005); and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3): 185-91 (2005)).Human antibodies can also be generated by isolating selected Fv clone variable domain sequences from human-derived phage presentation libraries. These variable domain sequences can then be combined with a desired human constant region. Oligosaccharide modifications in an antibody can be made, for example, to create antibody variants with certain enhanced properties. For instance, glycosylated antibody variants may have enhanced CDC function. In some embodiments, the present description may include an antibody variant that possesses some, but not all, effector functions, making it a suitable candidate for applications where the in vivo antibody half-life is important, but certain effector functions (such as complement) are unnecessary or detrimental. In vitro and / or in vivo cytotoxicity assays can be performed to confirm the reduction / depletion of CDC activities. In some embodiments, an antibody agent provided herein may be further modified to contain additional non-protein residues known in the art and readily available. Suitable residues for obtaining the antibody may include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers may include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, and alcohol. MA / a / ZUZl / Ul dUUO polyvinyl, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acids (homopolymers or random copolymers) and dextran or poly(n-vinylpyrrolidone) polyethylene glycol, polypropylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., 5-glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may have manufacturing advantages due to its stability in water. The polymer can have any molecular weight and can be branched or unbranched. The number of polymers bound to the antibody can vary, and if two or more polymers are bound, they can be the same or different molecules. In some formulations, conjugates of an antibody and a non-protein moiety are provided that can be selectively heated by exposure to radiation. In some formulations, the non-protein moiety may be a carbon nanotube (see, for example, Kam et al., Proc. Nati. Acad. Sci. USA, 102: 11600-11605 (2005)). The radiation may be of any wavelength and may include, but is not limited to, wavelengths that do not damage normal cells but heat the non-protein moiety to a temperature at which cells near the antibody's non-protein moiety die. b. Chimeric antigen receptors In some embodiments, a manipulated immune cell comprises a population of CARs, each CAR comprising an extracellular antigen-binding domain. In some embodiments, an immune cell comprises a population of CARs, each CAR comprising the same extracellular antigen-binding domains. As used in this description, chimeric antigen receptors (CARs) are proteins that specifically recognize target antigens (e.g., target antigens on 25 cancer cells). When bound to the target antigen, the CAR can activate the immune cell to attack and destroy the cell bearing that antigen (e.g., the cancer cell). CARs may also incorporate costimulatory or signaling domains to increase their potency. See Krause et al., J. Exp. Med., Vol. 188, No. 4, 1998 (619-626); Finney et al., Journal of Immunology, 1998, 161: 2791-2797; Song et al., Blood 119:696-706 (2012); Kalos et al., Sci. Trans. Med. 3:95 (2011); Porter et al., N. Engl. J. Med. 365:725-33 (2011) and Gross et al., Annu. Rev. Pharmacol. Toxico!. 56:59-83 (2016); U.S. patents nos. 7,741,465 and 6,319,494. The chimeric antigen receptors described herein comprise an extracellular domain, a transmembrane domain, and an intracellular domain, wherein the extracellular domain 35 comprises an antigen-binding domain that binds specifically to the target. In some forms, antigen-specific CARs also include a safety switch and / or one or more monoclonal antibody-specific epitopes. i. Antigen-binding domains As discussed previously, the CARs described herein comprise an antigen-binding domain. An antigen-binding domain, as used herein, means any polypeptide that binds to a specific target antigen. In some embodiments, the antigen-binding domain binds to an antigen on a tumor cell. In some embodiments, the antigen-binding domain binds to an antigen on a cell involved in a hyperproliferative disease. In some embodiments, the antigen-binding domain comprises a variable heavy chain, a variable light chain, and / or one or more CDRs described herein. In some embodiments, the antigen-binding domain is a single-chain variable fragment (scFv), comprising light chain CDRs CDR1, CDR2, and CDR3, and heavy chain CDRs CDR1, CDR2, and CDR3. Antigen-binding domain variants (e.g., CDR, VH, and / or VL variants) are also within the scope of this description, for example, variable light and / or variable heavy chains, each having at least 70–80%, 80–85%, 85–90%, 90–95%, 95–97%, 97–99%, or greater than 99% identity with the amino acid sequences of the 20 antigen-binding domain sequences. In some cases, such molecules include at least one heavy chain and one light chain, while in other cases, the variant forms contain two variable light chains and two variable heavy chains (or subparts thereof). A skilled individual may determine the appropriate antigen-binding domain variants as described herein using 25 well-known techniques.In certain modalities, an expert in the technique can identify suitable areas of the molecule that can be changed without destroying activity by targeting regions not believed to be important for activity. In certain embodiments, the polypeptide structure of the antigen-binding domains is based on antibodies, including, but not limited to, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as antibody mimetics), chimeric antibodies, humanized antibodies, human antibodies, antibody fusions (sometimes referred to herein as antibody conjugates), and fragments thereof, respectively. In some embodiments, the antigen-binding domain comprises or consists of avimers. An antigen-binding domain is said to be selective when it binds to one target more strongly than it binds to a second target. In some modality, an antigen-binding domain is an scFv. In some forms, an antigen-selective CAR comprises a leader or signal peptide. In other embodiments, the description refers to isolated polynucleotides encoding any of the antigen-binding domains described herein. In some embodiments, the description refers to isolated polynucleotides encoding a CAR. Vectors comprising the polynucleotides and methods for preparing them are also provided herein. ii. Safety switches and specific epitopes of monoclonal antibodies It will be appreciated that adverse events can be minimized by transducing immune cells (containing one or more CARs) with a suicide gene. It may also be desirable to incorporate an inducible on / off switch or accelerator into the immune cells. Suitable techniques include the use of inducible caspase-9 (request for United States 2011 / 0286980) or a thymidine kinase, before, after, or at the same time, when cells are transduced with the CAR construct described herein. Additional methods for introducing suicide genes and / or on-switches include TALENS, zinc fingers, iRNA, siRNA, hsRNA, antisense technology, and 20 other techniques known in the art. According to the description, other on / off switching techniques or other types of control may be incorporated herein. These techniques may employ the use of dimerization domains and optional actuators of such dimerization domains. These techniques include, for example, those described by Wu et al., Science 2014 350 (6258), which utilize FKBP / Rapalog dimerization systems in certain cells, the contents of which are incorporated herein by reference in their entirety. Additional dimerization technology is described in, for example, Fegan et al., Chem. Rev. 2010, 110, 33153336, as well as in U.S. Patent Nos. 5,830,462; 5,834,266; 5,869,337; and 6,165,787, the contents of which are also incorporated herein by reference in their entirety.Additional dimerization pairs may include cyclosporine A / cyclophilin, receptor, estrogen / estrogen receptor (optionally by using tamoxifen), glucocorticoid / glucocorticoid receptor, tetracycline / tetracycline receptor, vitamin D / vitamin D receptor. Additional examples of dimerization technology can be found in, for example, documents WO 2014 / 127261, WO 2015 / 090229, US 35 2014 / 0286987, US2015 / 0266973, US2016 / 0046700, US patents no. 8,486,693, US 2014 / 0171649 and US 2012 / 0130076, the contents of which are additionally incorporated in this description by reference in their entirety. In some embodiments, the CAR immune cell (e.g., CAR-T cell) described herein comprises a polynucleotide encoding a suicide polypeptide, such as RQR8. See, for example, WO2013153391 A, which is hereby incorporated by reference in its entirety. In CAR immune cells (e.g., CAR-T cells) comprising the polynucleotide, the suicide polypeptide is expressed on the surface of a CAR immune cell (e.g., CAR-T cell). In some embodiments, the suicide polypeptide comprises the amino acid sequence shown in SEQ ID NO: 1. CPYSNPSLCSGGGGSELPTQGTFSNVSTNVSPAKPTTTACPYSNPSLCSGGGGSP APPRPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLS LVITLYCNHRNRRRVCKCPRPVV (SEQ ID NO: 1). The suicide polypeptide may further comprise a signal peptide at the amino terminus, e.g., MGTSLLCWMALCLLGADHADA (SEQ ID NO: 2). In some embodiments, the suicide polypeptide comprises the amino acid sequence shown in SEQ ID NO: 3, which includes the signal sequence of SEQ ID NO: 2. MGTSLLCWMALCLLGADHADACPYSNPSLCSGGGGSELPTQGTFSNVSTNVSPAKPTTTAC PYSNPSLCSGGGGSPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPL AGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVV (SEQ ID NO: 3). When the suicide polypeptide is expressed on the surface of a CAR immune cell (e.g., CAR-T cell), the binding of rituximab to the R epitopes of the polypeptide causes cell lysis. More than one rituximab molecule can bind to each polypeptide expressed on the cell surface. Each R epitope of the polypeptide can bind to a separate rituximab molecule. Deletion of antigen-specific CAR immune cells (e.g., CAR-T cells) can occur in vivo, for example, by administering rituximab to a patient. The decision to remove the transferred cells may arise from the detection of undesirable effects in the patient that are attributable to the transferred cells, such as when unacceptable levels of toxicity are detected. In some modality, a suicide polypeptide is expressed on the cell surface. In some modality, a suicide polypeptide is included in the CAR construct. In some modality, a suicide polypeptide is not part of the CAR construct. In some embodiments, the extracellular domain of an antigen-specific CAR may comprise one or more epitopes specific to (i.e., specifically recognized by) a monoclonal antibody. These epitopes are also referred to herein as mAb-specific epitopes. Illustrative mAb-specific epitopes are described in International Patent Publication No. WO 2016 / 120216, which is incorporated herein in its entirety. In these embodiments, the extracellular domain of the CARs comprises antigen-binding domains that bind specifically to an antigen and one or more epitopes that bind to one or more monoclonal antibodies (mAbs). The CARs comprising the mAb-specific epitopes may be single-stranded or multi-stranded.The inclusion of monoclonal antibody-specific epitopes in the extracellular domain of the CARs described herein allows for the sorting and depletion of the engineered immune cells expressing the CARs. In some modalities, this feature also promotes the recovery of cells expressing endogenous antigens that were depleted by the administration of engineered immune cells expressing the CARs. In some modalities, allowing for depletion provides a safety mechanism in case of adverse effects, for example, following administration to a subject. Accordingly, in some modalities, the present description refers to a method for classifying and / or depleting manipulated immune cells equipped with CARs comprising specific mAb epitopes and a method for promoting the recovery of cells expressing endogenous antigens. Several monoclonal antibody-epitope pairs can be used to generate CARs comprising monoclonal antibody-specific epitopes; in particular, those already approved for medical use, such as the CD20 / rituximab epitope as a non-limiting example. The description also includes methods for classifying engineered immune cells equipped with antigen-specific CARs that express the specific mAb epitope(s) and therapeutic methods in which the activation of engineered immune cells equipped with these CARs is modulated by cell exhaustion using an antibody that targets the external ligand-binding domain of these CARs. Table 1 provides illustrative mimotope sequences that can be inserted into the extracellular domains of any of the CARs described. MA / a / ZUZl / Ul dUUO Table 1. Illustrative mimotope sequences Rituximab Mimotopo SEQ ID NO: 4 CPYSNPSLC Palivizumab Epitope SEQ ID NO: 5 NSELLSLINDMPITNDQKKLMSNN Cetuximab Mimotopo 1 SEQ ID NO: 6 CQFDLSTRRLKC Mimotopo 2 SEQ ID NO: 7 CQYLSTRLKC Mimotope SEQ ID NO: 7 CVWQRWQKSYVC Mimótopo 4 SEQ ID NO: 9 CMWDRFSRWYKC Nivolumab Epitope 1 SEQ ID NO: 10 SFVLNWYRMSPSNQTDKLAAFPEDR Epitope 2 SEQ ID NO: 11 SGTYLCGAISLAPKAQIKE Qí 10 EBEND-12 SEQ ID NO: 12 ELPTQGTFSNVSTNVS Epitope 2 SEQ ID NO: 25 ELPTQGTFSNVSTNVSPAKPTTTA Alemtuzumab Epitope SEQ ID NO: 13 GQNDTSQTSSPS In some embodiments, the extracellular binding domain of the CAR comprises the following sequence Vi-Li-V2-(L)x-Epitope1-(L)x-; Vi-Li-V2-(L)x-Epitope1-(L)x-Epitope2-(L)x-; Vi-Li-V2-(L)x-Epitope1 -(L)x-Epitope2-(L)x-Epitope3-(L)x-; (L)x-Epitope1-(L)x-Vi-Li-V2; (L)x-Epitope1-(L)x-Epitope2-(L)x-Vi-L1-V2; Epitope1-(L)x-Epitope2-(L)x-Epitope3-(L)x-Vi-Li-V2; (L)x-Epítopo1-(L)x-Vi-Li-V2-(L)x-Epítopo2-(L)x; (L)x-Epítopo1-(L)x-Vi-Li-V2-(L)x-Epítopo2-(L)x-Epítopo3-(L)x-; (L)x-Epítopo1-(L)x-Vi-Li-V2-(L)x-Epítopo2-(L)x-Epítopo3-(L)x-Epítopo4-(L)x-; (L)x-Epítopo1-(L)x-Epítopo2-(L)x-Vi-L1-V2-(L)x-Epítopo3-(L)x-; (L)x-Epítopo1-(L)x-Epítopo2-(L)x-Vi-L1-V2-(L)x-Epítopo3-(L)x-Epítopo4-(L)x-; Vi-(L)x-Epítopo1-(L)x-V2; Vi-(L)x-Epítopo1-(L)x-V2-(L)x-Epítopo2-(L)x; Vi-(L)x-Epítopo1-(L)x-V2-(L)x-Epítopo2-(L)x-Epítopo3-(L)x; Vi-(L)x-Epítopo1-(L)x-V2-(L)x-Epítopo2-(L)x-Epítopo3-(L)x-Epítopo4-(L)x; (L)x-Epítopo1-(L)x-Vi-(L)x-Epítopo2-(L)x-V2; O, (L)x-Epítopo1-(L)x-Vi-(L)x-Epítopo2-(L)x-V2-(L)x-Epítopo3-(L)x; where, Vi es Vl y V2es Vh or Vi es Vh y V2es Vl; There is a suitable tightener to join the Vh chain to the Vl chain; Epitope 1, epitope 2, epitope 3, and epitope 4 are mAb-specific epitopes and may be identical or different, where Vh is a heavy chain variable fragment and Vl is a light chain variable fragment. iii. Hinge domain The extracellular domain of the CARs described may include a hinge domain (or hinge region). The term generally refers to any polypeptide that functions to link the transmembrane domain of a CAR to the extracellular antigen-binding domain. In particular, hinge domains can be used to provide greater flexibility and accessibility to the extracellular antigen-binding domain. A hinge domain can comprise up to 300 amino acids, in some forms from 10 to 100 amino acids, or in some forms from 25 to 50 amino acids. The hinge domain can be derived from all or part of natural molecules, such as all or part of the extracellular region of CD8, CD4, CD28, 4-1BB, or IgG (in particular, the hinge region of a IgG; it will be noted that the hinge region may contain part or all of a member of the immunoglobulin family, such as IgG1, IgG2, IgG3, IgG4, IgA, IgD, IgE, IgM, or a fragment thereof, or all or part of an antibody heavy chain constant region. Alternatively, the A domain may be a synthetic sequence that corresponds to a natural A sequence, or it may be a completely synthetic A sequence. In some embodiments, the A domain is part of the human CD8a chain (e.g., NP_001139345.1). In another embodiment, the hinge and transmembrane domains comprise a portion of the human CD8a chain. In some embodiments, the hinge domain of the CARs described herein comprises a subsequence of CD8o, an IgG1, IgG4, PD-1, or FcyRIIIa, specifically the hinge region of any of a CD8o, an IgG1, an IgG4, PD-1, or FcyRIIIa. In some embodiments, the hinge domain comprises a human CD8a hinge, a human IgG1 hinge, a human IgG4 hinge, a human PD-1 hinge, or a human FcyRIIIa hinge. In some modalities, the CARs described in the present description comprise a human scFv, hinge domains and 35 transmembrane CD8a, the CD3ζ signaling domain and a 4-1BB signaling domain.Table 2 provides illustrative hinge amino acid sequences provided in this description. Table 2. Illustrative hinges 5 Domain Amino acid sequence SEQ ID NO: FcyRIIIa hinge GLAVSTISSFFPPGYQ ​​14 10 CD8a hinge TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD 15 15 LGG1 hinge EPKSPDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMIARTPEVTCV VDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK 16 iv. Transmembrane domain The CARs described here are designed with a transmembrane domain that is fused to the extracellular domain of the CAR. Similarly, it can be fused to the intracellular domain of the CAR. In some cases, the transmembrane domain can be selected or modified by amino acid substitution to prevent such domains from binding to the transmembrane domains of identical or different surface membrane proteins, thereby minimizing interactions with other members of the receptor complex. In some embodiments, short linkers can form links between any or some of the extracellular, transmembrane, and intracellular domains of the CAR. The transmembrane domains suitable for a CAR described herein have the ability to (a) be expressed on the surface of an immune cell such as, for example, without limitation, a lymphocyte, such as a helper T cell (Th), cytotoxic T cell (Tc), regulatory T cell (Treg), or natural killer (NK) cells, and / or (b) interact with the extracellular antigen-binding domain and the intracellular signaling domain to direct the cellular response of an immune cell against a target cell. The transmembrane domain can be derived from a natural or synthetic source. When the source is natural, the domain can be derived from any transmembrane or membrane-bound protein. The transmembrane regions of particular use in this description may be derived from (comprising or corresponding to) CD28, OX-40, 4-1BB / CD137, CD2, CD7, CD27, CD30, CD40, programmed cell death 1 (PD-1), inducible T-cell costimulatory factor (ICOS), lymphocyte function-associated antigen 1 (LFA-1, CD1-1a / CD18), CD3 gamma, CD3 delta, CD3 epsilon, CD247, CD276 (B7-H3), LIGHT, (TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fe receptor gamma, MHC class 1 molecule, TNF receptor proteins, an immunoglobulin protein, cytokine receptor, integrins, lymphocyte activation signaling molecules (SLAM proteins), activating NK cell receptors, BTLA, a ligand receptor Toll, ICAM-1, B7-H3, CDS, ICAM-1, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL-2R beta, IL-2R gamma, IL-7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD1 1 d, ITGAE, CD103, ITGAL, CD1 1a, LFA-1, ITGAM,CD1 1b, ITGAX, CD1 1c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRT AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, a ligand that binds specifically with CD83 or cualquiera de sus combinaciones., As non-limiting examples, the transmembrane region may be derived from, or be a portion of, a T-cell receptor such as the α, β, γ, or δ polypeptide that constitutes the CD3 complex, p55 (one chain), p75 (β chain), or IL-2 receptor chain, Fe receptor subunit chain, particularly the Fcy receptor III, or CD proteins. Alternatively, the transmembrane domain may be synthetic and may predominantly comprise hydrophobic residues such as leucine and valine. In some embodiments, such a transmembrane domain is derived from the human CD8a chain (e.g., NP 001139345.1). In some embodiments, the transmembrane domain in the CAR of the description is a CD8a transmembrane domain. In some embodiments, the transmembrane domain in the CAR of the description is a CD8a transmembrane domain comprising the amino acid sequence IYIWAPLAGTCGVLLLSLVIT (SEQ ID NO: 17). In some embodiments, the CD8a transmembrane domain comprises the nucleic acid sequence encoding the transmembrane amino acid sequence of SEQ ID NO: 17. In some embodiments, the hinge and transmembrane domain in the CAR of the description is a CD8a hinge and a transmembrane domain comprising the amino acid sequence of SEQ ID NO: 17. In some embodiments, the transmembrane domain in the CAR of the description is a CD28 transmembrane domain. In some embodiments, the transmembrane domain in the CAR of the description is a CD28 transmembrane domain comprising the amino acid sequence FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 18). In some embodiments, the CD28 transmembrane domain comprises the nucleic acid sequence encoding the transmembrane amino acid sequence SEQ ID NO: 18. v. Intracellular domain The intracellular (cytoplasmic) domain of the CARs described can provide activation of at least one of the normal effector functions of the immune cell comprising the CAR. The effector function of a T cell, for example, may refer to cytolytic activity or auxiliary activity, which includes cytokine secretion. In some modalities, an activation intracellular signaling domain for use in a CAR may be the cytoplasmic sequences of, for example, without limitation, the T cell receptor and coreceptors that work together to initiate signal transduction after antigen receptor binding, as well as any derivative or variant of these sequences and any synthetic sequence having the same functional capability. It will be appreciated that the appropriate intracellular domains (e.g., activators) include, but are not limited to, signaling domains derived from (or corresponding to) CD28, OX40, 4-1BB / CD137, CD2, CD7, CD27, CD30, CD40, programmed cell death 1 (PD-1), inducible T-cell costimulatory factor (ICOS), lymphocyte function-associated antigen 1 (LFA-1, CD11a / CD18), CD3 gamma, CD3 delta, CD3 epsilon, CD247, CD276 (B7-H3), LIGHT, (TNFSF14), 20NKG2C, Ig alpha (CD79a), DAP-10, Fe receptor gamma, MHC class 1 molecule, TNF receptor proteins, an immunoglobulin protein, cytokine receptor, integrins, lymphocyte activation signaling molecules (proteins SLAM), activating NK cell receptors, BTLA, Toll ligand receptors, ICAM-1, B7-H3, CDS, ICAM-1, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL-2R beta, IL-2R gamma, IL-7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6,CD49f, ITGAD, CD1 1d, ITGAE, CD103, ITGAL, CD1 1a, LFA-1, ITGAM, CD1 1b, ITGAX, CD1 1c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRT AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, 30 Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, a ligand that binds specifically to CD83 or any of its combinations. The intracellular domains of the CARs described herein may incorporate, in addition to the activation domains described above, costimulatory signaling domains 35 (referred to interchangeably in this description as costimulatory molecules) to increase their potency. The costimulatory domains may provide a signal in addition to the primary signal provided by an activation molecule as described herein. It will be appreciated that the appropriate costimulatory domains within the scope of the description may be derived from (or correspond to), for example, CD28, OX40, 4-1BB / CD137, CD2, CD3 (alpha, beta, delta, epsilon, gamma, zeta), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, PD-1, ICOS, lymphocyte function-associated antigen 1 (LFA-1 (CD11a / CD18)), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fe gamma receptor, MHC class I molecule, TNFR, integrin, lymphocytic activation signaling molecule, BTLA, Toll ligand receptors, ICAM-1, B7-H3, CDS, ICAM-1, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL-2R beta, IL-2R gamma, IL-7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD1-1d, ITGAE, CD103, ITGAL, CD1-1a, LFA-1, ITGAM,CD1 -1b, ITGAX, CD1-1c, ITGB1,CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRT AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand or fragments or their combinations. It will be noted that additional costimulatory molecules, or fragments thereof, not listed above are within the scope of the description. In some embodiments, the intracellular / cytoplasmic domain of the CAR can be designed to comprise the 4-1BB / CD137 domain as such or combined with any other desired intracellular domain(s) useful in the context of the described CAR. The complete native amino acid sequence of 4-1BB / CD137 is described in NCBI reference sequence NP_001552.2. The complete native nucleic acid sequence of 4-1BB / CD137 is described in NCBI reference sequence NM_001561.5. In some embodiments, the intracellular / cytoplasmic domain of the CAR can be designed to comprise the CD28 domain as such or combined with any other desired intracellular domain(s) useful in the context of the CAR described. The complete native amino acid sequence of CD28 is described in NCBI reference sequence NP 006130.1. The complete native nucleic acid sequence of CD28 is described in NCBI reference sequence NM_006139.1. In some embodiments, the intracellular / cytoplasmic domain of the CAR can be designed to comprise the CD3 zeta domain as such or combined with any other desired intracellular domain(s) useful in the context of the CAR described. MA / a / ZUZl / Ul dUUO In some modalities, the intracellular signaling domain of the CAR may comprise the Οϋ3ζ signaling domain having an amino acid sequence with at least approximately 70%, at least 80%, at least 90%, 95%, 97%, or 99% sequence identity with an amino acid sequence shown in SEQ ID NO: 19 or SEQ ID NO: 26. LRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLY NELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (SEQ ID NO: 19) LRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLY NELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (SEQ ID NO: 26) For example, the intracellular domain of the CAR may comprise a portion of the CD3 zeta chain and a portion of a costimulatory signaling molecule. The intracellular signaling sequences within the intracellular signaling portion of the CAR described may be joined together in a random or specified order. In some embodiments, the intracellular domain is designed to comprise the CD3 zeta activation domain and a CD28 signaling domain. In some embodiments, the intracellular domain is designed to comprise the CD3 zeta activation domain and a 4-1 BB signaling domain. In some forms, the 4-1 BB (intracellular domain) comprises the amino acid sequence KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL (SEQ ID NO: 20). In some modalities, the 4-1 BB (intracellular domain) is encoded by the nucleic acid sequence: AAGCGCGGCAGGAAGAAGCTCCTCTACATTTTTAAGCAGCCTTTTATGAGGCCCGTACAG ACAACACAGGAGGAAGATGGCTGTAGCTGCAGATTTCCCGAGGAGGAGGAAGGTGGGTG CGAGCTG (SEQ ID NO: 21). In some modalities, the intracellular domain in the CAR is designed to comprise a portion of CD28 and CD3 zeta, wherein the intracellular CD28 comprises the nucleic acid sequence exposed in SEQ ID NO: 22. AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCC ACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCG CTGCCTATCGGAGC (SEQ ID NO: 22). In some embodiments, the intracellular domain in the CAR is designed to comprise the amino acid sequence RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS (SEQ ID NO: 23). The CD3 zeta amino acid sequence may comprise SEQ ID NO: 23, and the nucleic acid sequence may comprise SEQ ID NO: 24. AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAAC CAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCG CAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGT CTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAA GGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTA CGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 24). In some embodiments, the intracellular signaling domain of the described CAR comprises a domain of a costimulatory molecule. In some embodiments, the intracellular signaling domain of a described CAR comprises a portion of the costimulatory molecule selected from the group consisting of a fragment of 4-1BB (GenBank: AAA53133.) and CD28 (NP_006130.1). In some embodiments, the intracellular signaling domain of the described CAR comprises an amino acid sequence comprising at least 70%, at least 80%, at least 90%, 95%, 97%, or 99% sequence identity with an amino acid sequence shown in SEQ ID NO: 20 and SEQ ID NO: 22. In some embodiments, the intracellular signaling domain of the CAR described comprises an amino acid sequence comprising at least 70%, at least 80%, at least 90%, 95%, 97%, or 99% sequence identity with an amino acid sequence shown in SEQ ID NO: 20 and / or at least 70%, at least 80%, at least 90%, 95%, 97%, or 99% sequence identity with an amino acid sequence shown in SEQ ID NO: 22. c. Immune cells comprising CAR In this description, manipulated immune cells expressing the CARs described are provided (e.g., CAR-T cells). In some embodiments, a manipulated immune cell comprises a population of CARs, each CAR comprising different extracellular antigen-binding domains. In some embodiments, a manipulated immune cell comprises a population of CARs, each CAR comprising the same extracellular antigen-binding domains. Manipulated immune cells can be allogeneic or autologous. In some modalities, the manipulated immune cell is a T cell (e.g., inflammatory T lymphocyte, cytotoxic T lymphocyte, regulatory T lymphocyte, helper T lymphocyte, tumor-infiltrating lymphocyte (TIL)), NK cell, NK-T cell, TCR-expressing cell, dendritic cell, killer dendritic cell, mast cell, or B cell. In some modalities, the cell may be derived from the group consisting of CD4+ T lymphocytes and CD8+ T lymphocytes. In some illustrative modalities, the manipulated immune cell is a T cell. In some illustrative modalities, the manipulated immune cell is a gamma delta T cell. In some illustrative modalities, the manipulated immune cell is a macrophage. In some modalities, the manipulated immune cell can be derived from, for example, without limitation, a stem cell. Stem cells can be adult stem cells, non-human embryonic stem cells, more particularly non-human stem cells, umbilical cord blood stem cells, progenitor cells, bone marrow stem cells, induced pluripotent stem cells, totipotent stem cells, or hematopoietic stem cells. In some modality, the cell is obtained or prepared from peripheral blood. In some modality, the cell is obtained or prepared from peripheral blood mononuclear cells (PBMCs). In some modality, the cell is obtained or prepared from bone marrow. In some modality, the cell is obtained or prepared from umbilical cord blood. In some modality, the cell is a human cell. In some modality, the cell is transfected or transduced with the nucleic acid vector using a method selected from the group consisting of electroporation, sonoporation, biolistics (e.g., gene gun), lipid transfection, polymer transfection, nanoparticles, viral transfection (e.g., retrovirus, lentivirus, AAV), or polyplexes. In some modalities, the manipulated immune cells that express on their cell surface membrane a CAR specific to the antigen described comprise a percentage of stem cell and central memory cells greater than 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. In some modalities, the manipulated immune cells expressing on their cell surface membrane a CAR specific to the antigen described comprise a percentage of stem cell and core memory cells of approximately 10% to approximately 100%, approximately 10% to approximately 90%, approximately 10% to approximately 80%, approximately 10% to approximately 70%, approximately 10% to approximately 60%, approximately 10% to approximately 50%, approximately 10% to approximately 40%, approximately 10% to approximately 30%, approximately 10% to approximately 20%, approximately 15% to approximately 100%, approximately 15% to approximately 90%, approximately 15% to approximately 80%, approximately 15% to approximately 70%, approximately 15% to approximately 60%, approximately 15% to approximately 50%, approximately 15% to approximately 40%,approximately 15% to approximately 30%, approximately 20% to approximately 100%, approximately 20% to approximately 90%, approximately 20% to approximately %, approximately 20% to approximately 70%, approximately 20% to approximately 60%, approximately 20% to approximately 50%, approximately 20% to approximately 40%, approximately 20% to approximately 30%, approximately 30% to approximately 100%, approximately 30% to approximately 90%, approximately 30% to approximately 80%, approximately 30% to approximately 70%, approximately 30% to approximately 60%, approximately 30% to approximately 50%, approximately 30% to approximately 40%, approximately 40% to approximately 100%, approximately 40% to approximately 90%, approximately 40% to approximately 80%, approximately 40% to approximately 70%, approximately 40% to approximately 60%, approximately 40% to approximately 50%,approximately 50% to approximately 100%, approximately 50% to approximately 90%, approximately 50% to approximately 80%, approximately 50% to approximately 70%, approximately 50% to approximately 60%, approximately 60% to approximately 100%, approximately 60% to approximately 90%, approximately 60% to approximately 80%, approximately 60% to approximately 70%, approximately 70% to approximately 90%, approximately 70% to approximately 80%, approximately 80% to approximately 100%, approximately 80% to approximately 90%, approximately 90% to approximately 100%, approximately 25% to approximately 50%, approximately 75% to approximately 100%, or approximately 50% to approximately 75%. In some modalities, the manipulated immune cells that express on their cell surface membrane a CAR specific to the antigen described comprise a percentage of stem cell and central memory cells greater than 10%, 20%, 30%, 40%, 50%, or 60%. In some modalities, the manipulated immune cells expressing on their cell surface membrane a CAR specific to the antigen described comprise a percentage of stem cell and core memory cells of approximately 10% to approximately 60%, approximately 10% to approximately 50%, approximately 10% to approximately 40%, approximately 15% to approximately 50%, approximately 15% to approximately 40%, approximately 20% to approximately 60%, or approximately 20% to approximately 70%. In some embodiments, the engineered immune cells expressing an antigen-specific CAR on their cell surface membrane, as described, are enriched in Tcm and / or Tscm cells such that the engineered immune cells comprise at least approximately 60%, 65%, 70%, 75%, or 80% of combined Tcm and Tscm cells. In some embodiments, the manipulated immune cells expressing on their cell surface membrane a CAR specific to the antigen described are enriched in Tcm and / or Tscm cells such that the manipulated immune cells comprise at least approximately 70% of combined Tcm and Tscm cells. In some embodiments, the manipulated immune cells expressing on their cell surface membrane a CAR specific to the antigen described are enriched in Tcm and / or Tscm cells such that the manipulated immune cells comprise at least approximately 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% or 60% Tscm cells. In some modalities, the immune cell is an inflammatory T lymphocyte expressing any of the CARs described herein. In some modalities, the immune cell is a cytotoxic T lymphocyte expressing any of the CARs described herein. In some modalities, the immune cell is a regulatory T lymphocyte expressing any of the CARs described herein. In some modalities, the immune cell is a helper T lymphocyte expressing any of the CARs described herein. This description also provides cell lines obtained from a transformed immune cell (e.g., T cell) according to any of the methods described above. Modified cells resistant to immunosuppressive treatment are also provided herein. In some embodiments, an isolated cell according to the description comprises a polynucleotide encoding a CAR. In some embodiments, an immune cell manipulated according to the present description may comprise one or more disrupted or inactivated genes. In some embodiments, an immune cell manipulated according to the present description comprises a disrupted or inactivated gene selected from the group consisting of CD52, DLL3, GR, PD-1, CTLA-4, LAG3, TIM3, BTLA, BY55, TIGIT, B7H5, LAIR1, SIGLEC10, 2B4, HLA, TCRa, and TCRp and / or expresses the transgene of a CAR, a multi-stranded CAR, and / or a pTa. In some embodiments, an isolated cell comprises polynucleotides encoding polypeptides comprising a multi-stranded CAR.In some embodiments, the isolated cell according to the present description comprises two interrupted or inactivated genes selected from the group consisting of: CD52 and GR, CD52 and TCRa, CDR52 and TCRp, DLL3 and CD52, DLL3 and TCRa, DLL3 and TCRp, GR and TCRa, GR and TCRp, TCRa and TCRp, PD-1 and TCRa, PD-1 and TCRp, CTLA-4 and TCRa, CTLA-4 and TCRp, LAG3 and TCRa, LAG3 and TCRp, TIM3 and TCRa, Tim3 and TCRp, BTLA and TCRa, BTLA and TCRp, BY55 and TCRa, BY55 and TCRp, TIGIT and TCRa, TIGIT and TCRp, B7H5 and TCRa, B7H5 and TCRp, LAIR1 and TCRa, LAIR1 and TCR3, SIGLEC10 and TCRa, SIGLEC10 and TCRp, 2B4 and TCRa, 2B4 and TCRP and / or expresses the transgene of a CAR, a multistranded CAR and a pTa. In some modalities, the method comprises disrupting or inactivating one or more genes by introducing into the cells an endonuclease that can selectively inactivate a gene by selective DNA excision.In some forms, the endonuclease may be, for example, a zinc finger nuclease (ZFN), megaTAL nuclease, meganuclease, transcription activator-type effector nuclease (TALE nuclease), or CRIPR endonuclease (e.g., Cas9). In some embodiments, the TCR is rendered non-functional in cells as described by disruption or inactivation of the TCRα gene and / or the TCRκ gene(s). In some embodiments, a method is provided for obtaining modified cells derived from an individual, wherein the cells can proliferate independently of the major histocompatibility complex (MHC) signaling pathway. Modified cells, capable of proliferating independently of the MHC signaling pathway and amenable to obtaining them by this method, are included within the scope of this description.The modified cells described herein can be used to treat patients in need of treatment against host-versus-graft (HvG) and graft-versus-host disease (GvHD); therefore, within the scope of this description is a method for treating patients in need of treatment against host-versus-graft (HvG) and graft-versus-host disease (GvHD) comprising treating such patient by administering to such patient an effective amount of modified cells comprising disrupted or inactivated TCRα and / or TCRβ genes. In some approaches, immune cells are engineered to be resistant to one or more chemotherapy drugs. The chemotherapy drug might be, for example, a purine nucleotide analog (PNA), making the immune cell suitable for cancer treatment that combines adoptive immunotherapy and chemotherapy. Illustrative PNAs include, for example, clofarabine, fludarabine, cyclophosphamide, and cytarabine, alone or in combination. Deoxycytidine kinase (dCK) metabolizes PNAs to mono-, di-, and triphosphate PNAs. Their triphosphate forms compete with ATP for DNA synthesis, act as pro-apoptotic agents, and are potent inhibitors of ribonucleotide reductase (RNR), which is involved in trinucleotide production. In some modalities, the isolated cells or cell lines described may comprise a pTo or a functional variant thereof. In some modalities, an isolated cell or cell line may be further genetically modified by disruption or inactivation of the TCRa gene. The description also provides for engineered immune cells comprising any of the CAR polynucleotides described herein. In some embodiments, a CAR can be introduced into an immune cell as a transgene using a plasmid vector. In some embodiments, the plasmid vector may also contain, for example, a selection marker that enables the identification and / or selection of cells that received the vector. CAR polypeptides can be synthesized in situ within the cell after the introduction of polynucleotides encoding the CAR polypeptides. Alternatively, CAR polypeptides can be produced outside of cells and then introduced into cells. Methods for introducing a polynucleotide construct into cells are known in the art. In some modalities, stable transformation methods (e.g., using a lentiviral vector) can be used to integrate the polynucleotide construct into the cell's genome. In other modalities, transient transformation methods can be used to transiently express the polynucleotide construct, and the polynucleotide construct is not integrated into the cell's genome. In still other modalities, virus-mediated methods can be used.Polynucleotides can be introduced into a cell by any suitable means, such as recombinant viral vectors (e.g., retroviruses, adenoviruses), liposomes, and the like. Transient transformation methods include, but are not limited to, microinjection, electroporation, or particle bombardment. Polynucleotides can be incorporated into vectors, such as plasmid vectors or viral vectors. In some embodiments, isolated nucleic acids are provided comprising a promoter operatively linked to a polynucleotide primer encoding an antigen-binding domain, at least one costimulatory molecule, and an activator domain. In some embodiments, the nucleic acid construct is contained within a viral vector. In some embodiments, the viral vector is selected from the group consisting of retroviral vectors, murine leukemia virus vectors, SFG vectors, adenoviral vectors, lentiviral vectors, adeno-associated virus (AAV) vectors, herpesvirus vectors, and vaccinia virus vectors. In some embodiments, the nucleic acid is contained within a plasmid. 3. Cell culture media for in vitro expansion of immune cells In vitro cell expansion (including the expansion of immune cells, such as genetically modified T cells) can be achieved using a cell culture medium comprising: a first stimulant of cell proliferation (e.g., a first cytokine that stimulates T cell proliferation); a second cell proliferation stimulant (e.g., a second cytokine that stimulates T cell proliferation); and an extracellular modulator of cell metabolism (e.g., T cell metabolism). The immune cells described herein can be activated and expanded either before or after genetic modification, using methods generally known and described herein. In some embodiments, the immune cells (e.g., T cells) are activated and expanded before genetic modification. In other embodiments, the immune cells (e.g., T cells) are activated and expanded after genetic modification (e.g., engineered immune cells, including those described herein). Accordingly, appropriate conditions for T cell expansion include a suitable medium (e.g., Essential Minimum Medium or RPMI 1640 medium, or X-vivo 15 (Lonza)) comprising the first and second cell proliferation stimulants (e.g., IL-7 and IL-15), as well as an extracellular modulator of cell metabolism (e.g., extracellular potassium), and may contain additional factors necessary for proliferation and viability, including serum (e.g., fetal bovine or human serum), IL-2, insulin, IFN-γ, IL-4, IL-7, GM-CSF, IL-10, IL-2, IL-15, TGFβ, and TNF, or any other cell growth additive known to the expert. In some modalities, the T cell culture medium does not include exogenous IL-2. Other cell growth additives include, but are not limited to, surfactant, plasmanate, and reducing agents such as N-acetylcysteine ​​and 2-mercaptoethanol.Media may include RPMI 1640, A1M-V, DMEM, MEM, α-MEM, F30 12, X-Vivo 15, and X-Vivo 20, Optimizer, with added amino acids, sodium pyruvate, and vitamins, either without serum or supplemented with an appropriate amount of serum (or plasma) or a defined set of hormones, and / or a sufficient amount of cytokine(s) for T-cell growth and expansion (e.g., IL-7 and / or IL-15). Antibiotics, e.g., penicillin and streptomycin, are included only in experimental cultures, not in cell cultures that will be infused into a subject. Target cells are maintained under the conditions necessary to support growth, e.g., an appropriate temperature (e.g., 37°C) and atmosphere (e.g., air plus 5% CO2). T cells that have been exposed to different stimulation times may exhibit different characteristics. The methods described herein may also include contacting cells with an agent that stimulates a CD3 TCR complex and a costimulatory molecule on the T cell surface to create an activation signal for the T cell. For example, chemicals such as calcium ionophore A23187, phorbol 12-myristate 13acetate (PMA), or mitogenic lectins such as phytohemagglutinin (PHA) may be used to create an activation signal for the T cell. In some modalities, T cell populations can also be stimulated in vitro by contact with, for example, an anti-CD3 antibody or an antigen-binding fragment thereof, or an anti-CD2 antibody immobilized on a surface, or by contact with a protein kinase C activator (e.g., bryostatin) along with a calcium ionophore. For co-stimulation of a helper molecule on the T cell surface, a ligand is used that binds to the helper molecule. For example, a T cell population can be contacted with an anti-CD3 antibody and an anti-CD28 antibody under conditions appropriate to stimulate T cell proliferation. The anti-CD3 antibody and the anti-CD28 antibody can be placed on a bead, plate, or other substrate. In some methods, the cells described can be expanded by co-culturing with tissue or other cells. The cells can also be expanded in vivo, for example, in the subject's bloodstream after administration. In particular, cell culture media and methods of using them can be particularly useful in the manufacture of T cells, including CAR-T cells such as 25 allogeneic CAR-T cells. a. Cell proliferation stimulants In some modalities, a first and a second stimulant are selected from: an agent that stimulates a CD3 TCR complex; an anti-CD3 antibody, or an antigen-binding fragment thereof; an anti-CD2 antibody or an antigen-binding fragment thereof; a protein kinase C activator; or a growth factor (e.g., a T cell growth factor) or any combination thereof. In some modalities, an immune cell stimulant (e.g., T cells or engineered immune cells) is an agent that stimulates a CD3 TCR complex and a costimulatory molecule on the surface of T cells to create an activation signal for the T cell. For example, chemicals such as calcium ionophore may be used. A23187, phorbol 12-myristate 13-acetate (PMA) or mitogenic lectins such as phytohemagglutinin (PHA) to create an activation signal for the T cell. In some formulations, an immune cell stimulant (e.g., T cells or engineered immune cells) is an anti-CD3 antibody, an antigen-binding fragment thereof, an anti-CD2 antibody immobilized on a surface, or a protein kinase C activator (e.g., bryostatin) along with a calcium ionophore. A ligand that binds to the T cell surface is used to co-stimulate an auxiliary molecule. For example, a population of T cells can be exposed to an anti-CD3 antibody and an anti-CD28 antibody under conditions appropriate for stimulating T cell proliferation. The anti-CD3 and anti-CD28 antibodies can be placed on a bead, plate, or other substrate. In some modalities, a cell stimulant is a growth factor. In some modalities, a cell stimulant is a T-cell growth factor. In some modalities, a T-cell growth factor is a cytokine. In some modalities, a first stimulant is a first cytokine and a second stimulant is a second cytokine. In some modalities, a cytokine is an interleukin (e.g., IL-2, IL-4, IL-7, IL-10, IL-12, or IL-15). In some modalities, a first and a second stimulant are selected from the group consisting of IL-4, IL-7, IL-10, IL-12, and IL-15. In some modalities, a first stimulant is IL-7 and a second stimulant is IL-15. In some modalities, a cell culture medium excludes IL-2 (e.g., a cell culture medium excludes exogenous IL-2 and IL-2 is not present in a cell culture medium as a first stimulant and / or second stimulant). In some modalities, the amounts of a first and second stimulant used are described in concentrations (e.g., using IU / ml). Therefore, a concentration ratio of x: / can be used to describe the ratio of the concentration of a first stimulant (e.g., x IU / ml of IL-7) to the concentration of a second stimulant (e.g., y IU / ml of IL-15). In some modalities, a first stimulant (e.g., a first cytokine such as IL7) and a second stimulant (e.g., a second cytokine such as IL-15) are present in a concentration ratio of approximately 10,000:1 to approximately 1:10,000, approximately 1,000:1 to approximately 1:1,000, approximately 100:1 to approximately 1:100, or approximately 10:1 to approximately 1:10. In some modalities, a first stimulant (e.g., a first cytokine such as IL7) is present in an amount that is greater than a second stimulant (e.g., a second cytokine such as IL-15). In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) are present in a concentration ratio of approximately 1,000:1 to approximately 10:1, approximately 900:1 to approximately 10:1, approximately 800:1 to approximately 10:1, approximately 700:1 to approximately 10:1, approximately 600:1 to approximately 10:1, approximately 500:1 to approximately 10:1, approximately 400:1 to approximately 10:1, approximately 300:1 to approximately 10:1, approximately 250:1 to approximately 10:1, approximately 200:1 to approximately 10:1, approximately 150:1 to approximately 10:1, or approximately 100:1 to approximately 4:1.In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) are present in a concentration ratio of approximately 1,000:1 to approximately 50:1, approximately 900:1 to approximately 50:1, approximately 800:1 to approximately 50:1, approximately 700:1 to approximately 50:1, approximately 600:1 to approximately 50:1, approximately 500:1 to approximately 50:1, approximately 400:1 to approximately 50:1, approximately 300:1 to approximately 50:1, approximately 250:1 to approximately 50:1, approximately 200:1 to approximately 50:1, approximately 150:1 to approximately 50:1, or approximately 100:1 to approximately 50:1.In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) are present in a concentration ratio of approximately 200:1 to approximately 10:1, approximately 150:1 to approximately 10:1, approximately 125:1 to approximately 10:1, or approximately 100:1 to approximately 4:1. In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) are present in a concentration ratio of approximately 100:1. In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) are present in a concentration ratio of approximately 5:1, approximately 6:1, or approximately 7:1.In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) are present in a concentration ratio of 6.25:1. In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) is present at a concentration of approximately 100 IU / ml to approximately 20,000 IU / ml. In some modalities, a second stimulant (e.g., a second cytokine such as IL-15) is present at a concentration of approximately 1 IU / ml to approximately 200 IU / ml. MA / a / ¿U¿l / Ul dUUO In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) is present at a concentration of approximately 100 IU / ml to approximately 10,000 IU / ml. In some modalities, a second stimulant (e.g., a second cytokine such as IL-15) is present at a concentration of approximately 1 IU / ml to approximately 100 IU / ml. In some modalities, a prime stimulant (e.g., a prime cytokine such as IL-7) is present at a concentration of approximately 100 IU / ml to approximately 1,000 IU / ml, approximately 300 IU / ml to approximately 5,000 IU / ml, or approximately 3,000 IU / ml. In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) is present at a concentration of approximately 2,500 IU / ml to approximately 7,500 IU / ml. In some modalities, a second stimulant (e.g., a second cytokine such as IL-15) is present at a concentration of approximately 25 IU / ml to approximately 75 IU / ml. In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) is present at a concentration of approximately 4,000 IU / ml to approximately 6,000 IU / ml. In some modalities, a second stimulant (e.g., a second cytokine such as IL-15) is present at a concentration of approximately 40 IU / ml to approximately 60 IU / ml. In some modalities, a second stimulant (e.g., a second cytokine such as IL-15) is present at a concentration of approximately 25 IU / ml to approximately 100 IU / ml or approximately 50 IU / ml. In some modalities, a first stimulant (e.g., a first cytokine such as IL-7) is present at a concentration of approximately 5,000 IU / ml. In some modalities, a second stimulant (e.g., a second cytokine such as IL-15) is present at a concentration of approximately 50 IU / ml. In some modalities, immune cells (e.g., T cells such as genetically modified T cells) are exposed to a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) approximately once every one, two, three, four, five, six, seven, or eight days. In some modalities, immune cells (e.g., T cells such as genetically modified T cells) are exposed to a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) approximately once every week.In some modalities, immune cells (e.g., T cells such as genetically modified T cells) are exposed to a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) once, twice, three times, four times, five times, six times, seven times, or eight times during an expansion process lasting approximately one day, two days, three days, four days, five days, six days, or seven days (e.g., approximately one week), approximately two weeks, approximately three weeks, or approximately four weeks. In some modalities, immune cells (e.g., T cells such as genetically modified T cells) are exposed to a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) for approximately two weeks. b. Extracellular modulators of cellular metabolism In some embodiments, a cell culture medium further comprises an extracellular modulator 15 of cell metabolism in addition to a first stimulant (e.g., a first cytokine such as IL-7) and a second stimulant (e.g., a second cytokine such as IL-15) as described herein. In some modalities, an extracellular modulator is extracellular potassium (K+). In some modalities, extracellular potassium is present at a concentration of approximately 20 mM to approximately 50 mM, approximately 4 mM to approximately 40 mM, approximately 4 mM to approximately 30 mM, approximately 10 mM to approximately 40 mM, approximately 10 mM to approximately 30 mM, approximately 15 mM to approximately 40 mM, approximately 15 mM to approximately 35 mM, approximately 15 mM to approximately 30 mM, or approximately 20 mM to approximately 30 mM. In some modalities, extracellular potassium is present at a concentration of approximately 10 mM to approximately 30 mM or approximately 20 mM to approximately 30 mM. In some modalities, extracellular potassium is present at a concentration that is approximately 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 30 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM or 30 mM. In some modalities, extracellular potassium is present at a concentration greater than approximately 15 mM. In some modalities, extracellular potassium is present at a concentration greater than or equal to approximately 20 mM. In some modalities, extracellular potassium is present at a concentration that is approximately 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, or 30 mM. MA / a / ZUZl / Ul dUUO In some modalities, extracellular potassium is present at a concentration of no less than approximately 10 mM. In some modalities, extracellular potassium is present at a concentration of no less than approximately 20 mM. In some modalities, extracellular potassium is present at a concentration of no more than approximately 30 mM. In some modalities, extracellular potassium is present at a concentration of approximately 25 mM. In some modalities, extracellular potassium is provided as KCI. c. Additional characteristics and components of cell culture media A cell culture medium as described herein (e.g., a cell culture medium comprising a first stimulant being IL-7, a second stimulant being IL-15, and an extracellular modulator being extracellular potassium) may be used in various methods for activating and expanding T cells known in the art and described, e.g., in U.S. Patent No. 6,905,874; U.S. Patent No. 6,867,041; United States patent no. 6,797,514; and the PCT document WO2012 / 079000, the contents of which are hereby incorporated by reference in their entirety. In some embodiments, a method further comprises contacting PBMCs or isolated T cells with an additional stimulatory molecule and a costimulatory molecule, such as anti-CD3 and anti-CD28 antibodies, generally bound to a bead or other surface, in a culture medium with appropriate cytokines, such as IL-2. The anti-CD3 and anti-CD28 antibodies bound to the same bead serve as a surrogate antigen-presenting cell (APC). An example is the Dynabeads® system, a CD3 / CD28 activator / stimulator system for the physiological activation of human T cells. In other embodiments, T cells can be activated and stimulated to proliferate with appropriate feeder cells and antibodies and still other cytokines (e.g., in addition to IL-7 and IL-15) by using methods such as those described in U.S. Patent No.6,040,177; U.S. Patent No. 5,827,642; and document WO2012129514, the contents of which are hereby incorporated by reference in their entirety. In some embodiments, the cell culture media and methods of use thereof exclude exogenous IL-2 as a T-cell growth factor. Other appropriate conditions for T cell culture include a suitable medium (e.g., Essential Minimum Medium or RPMI 1640 Medium or X-vivo™ Medium (Lonza)) that may contain factors in addition to, for example, the first and second cell proliferation stimulants, which include serum (e.g., fetal bovine or human serum 35), insulin, IFN-γ, GM-CSF, TGF-β, and TNF, or any other cell growth additive known to the expert. Other cell growth additives include, but are not limited to, surfactant, plasmanate, and reducing agents such as N-acetylcysteine ​​and 2-mercaptoethanol. The media may include RPM11640, A1M-V, DMEM, MEM, a-MEM, F-12, X-Vivo™ 10, X-Vivo™ 15 and X-Vivo™ 20, OpTmizer™, may contain added amino acids, sodium pyruvate and / or vitamins, and may not contain serum or be supplemented with an appropriate amount of serum (or plasma) and / or a defined set of hormones.Antibiotics such as penicillin and streptomycin are included only in experimental cultures, not in cell cultures that will be infused into a subject. The target cells are maintained under the conditions necessary to support growth, such as an appropriate temperature (e.g., 37°C) and atmosphere (e.g., air plus 5% CO2). T cells that have been exposed to different stimulation times may exhibit different characteristics In some methods, the cells described can be expanded by co-culturing with tissue or other cells. The cells can also be expanded in vivo, for example, in the subject's bloodstream after administration. In some forms, a cell culture medium contains serum. In some forms, a cell culture medium does not contain serum. In some forms, a cell culture medium does not contain components from other species. In some forms, a cell culture medium is chemically defined. d. Cell proliferation In some embodiments, the culture of cells (including immune cells such as T cells) for in vitro expansion using a cell culture medium described herein (e.g., a cell culture medium comprising a first stimulant being IL-7, a second stimulant being IL-15, and an extracellular modulator being extracellular potassium) or any of the methods described herein lasts for approximately one day, two days, three days, four days, five days, six days, seven days (e.g., approximately one week), approximately two weeks, approximately three weeks, or approximately four weeks. In some embodiments, the duration is from approximately one week to approximately three weeks or from approximately one week to approximately two weeks (e.g., approximately 7–15 days or approximately 10–14 days). In some embodiments, a cell culture medium described herein (e.g., a cell culture medium comprising a first stimulant being IL-7, a second stimulant being IL-15, and an extracellular modulator being extracellular potassium) or any of the methods described herein results in cell expansion of approximately 10 to approximately 10,000 times, measured over a period of approximately one week to approximately three weeks. In some embodiments, cell expansion of approximately 100 to approximately 1,000 times is observed over approximately 7–15 days (e.g., for approximately 10–14 days or approximately 2 weeks). In some embodiments, cell expansion of at least approximately 100 times is observed over approximately 7–15 days (e.g., for approximately 10–14 days or for approximately 2 weeks).In some modalities, cell expansion of at least approximately 200 times is observed for approximately 7-15 days (e.g., for approximately 10-14 days or for approximately 2 weeks). In some embodiments, a cell culture medium described herein (e.g., a cell culture medium comprising a first stimulant being IL-7, a second stimulant being IL-15, and an extracellular modulator being extracellular potassium) or any of the methods for cell expansion results in a cell count of at least approximately 50 × 10⁶ to approximately 100 × 10⁶ over a period of approximately 8–16 days. In some embodiments, the cell count is at least approximately 100 × 10⁶ over a period of approximately 10–14 days (e.g., at least approximately 100 × 10⁶, approximately 125 × 10⁶, approximately 150 × 10⁶, or approximately 100 × 10⁶ over a period of approximately 10–14 days). e. Cellular phenotypes In some embodiments, a cell culture medium described herein (for example, a cell culture medium comprising a first stimulant being IL-7, a second stimulant being IL-15, and an extracellular modulator being extracellular potassium) or any method of using the same produces a phenotype suitable for the production of 25 manipulated immune cells. In some embodiments, a T cell culture medium described herein (e.g., a cell culture medium comprising a first stimulant being IL-7, a second stimulant being IL-15, and an extracellular modulator being extracellular potassium) or any method of using the same produces a T cell population (e.g., genetically modified T30 cells) enriched in Tcm and Tscm cells. In some modalities, a T cell population (e.g., genetically modified T cells) enriched in Tcm and / or Tscm cells comprises at least approximately 60%, 65%, 70%, 75%, or 80% of combined Tcm and Tscm cells. In some modalities, a T cell population enriched in Tcm and / or Tscm cells comprises at least approximately 70% of combined Tcm and Tscm cells. In some modalities, a T cell population (e.g., genetically modified T cells) enriched in Tcm and Tscm cells comprises at least approximately 75% of combined Tcm and / or Tscm cells. In some modalities, a T cell population (e.g., genetically modified T cells) enriched in Tcm and / or Tscm cells comprises at least approximately 20.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% Tscm cells. In some modalities, a T cell population enriched in Tcm and / or Tscm cells comprises at least approximately 30%, 35%, 40%, or 45% Tscm cells. 4. Manufacturing of engineered immune cells (including CAR-T cells) This description provides methods of using cell culture media described herein for the manufacture of immune cells (including manipulated immune cells such as CAR-T cells). A variety of known techniques can be used to prepare the polynucleotides, polypeptides, vectors, antigen-binding domains, immune cells, compositions, and 15 likewise according to the description. Prior to the in vitro manipulation or genetic modification of the immune cells described herein, the cells may be obtained from a subject. The cells expressing a CAR may be derived from an allogeneic or autologous process. 20a. Source material The cell culture media described herein (e.g., comprising IL-7+IL-15 and increased extracellular potassium) can be used to culture various cells, including for the in vitro expansion of various immune cells (e.g., genetically modified T cells). Illustrative cells are described herein. In some modalities, immune cells comprise T cells. T cells can be obtained from various sources, including peripheral blood mononuclear cells (PBMCs), bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from an infection site, ascites, pleural effusion, spleen tissue, and tumors. In certain modalities, T cells can be obtained from a blood volume collected from the subject using any variety of techniques known to the practitioner, such as FICOLL™ separation. Cells can be obtained from an individual's circulating blood by apheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets. In some specific modalities, the cells collected by apheresis can be washed to remove the plasma fraction and then placed in an appropriate buffer or medium for further processing. In certain modalities, T cells are isolated from PBMCs by lysis of red blood cells and depletion of monocytes, for example, by centrifugation through a PERCOLL™ gradient. A specific T cell subpopulation (e.g., CD28+, CD4+, CD45RA-, and CD45RO+ T cells or CD28+, CD4+, CDS+, CD45RA-, CD45RO+, and CD62L+ T cells) can be further isolated using positive or negative selection techniques known in the technique. For example, enrichment of a T cell population by negative selection can be achieved with a combination of antibodies targeting unique surface markers for the negatively selected cells.One method for use in this description is cell sorting and / or selection by negative magnetic immunoadherence or flow cytometry using a cocktail of monoclonal antibodies targeting cell surface markers present on the negatively selected cells. For example, to enrich CD4+ cells by negative selection, a monoclonal antibody cocktail typically includes antibodies against CD14, CD20, CD11b, CD16, HLA-DR, and CD8. Flow cytometry and cell sorting can also be used to isolate cell populations of interest for use in this description. PBMCs can be used directly for genetic modification of immune cells (such as CAR or TCR cells) using methods as described herein. In certain modalities, after isolating the PBMCs, T lymphocytes can be further isolated, and cytotoxic and helper T lymphocytes can be sorted into naïve, memory, and effector T cell subpopulations before or after genetic modification and / or expansion. In some modalities, CD8+ cells are further classified as naïve, stem cell memory, central memory, and effector cells by identifying cell surface antigens associated with each of these CD8+ cell types. In some modalities, the expression of phenotypic markers of central memory T cells includes CD27, CD45RA, CD45RO, CD62L, CCR7, CD28, CD3, and CD127, and they are negative for granzyme B. In some modalities, stem cell memory T cells are CD45RO-, CD62L+, CD8+ T cells. In some modalities, central memory T cells are CD45RO+, CD62L+, CD8+ T cells. In some modalities, effector T cells are negative for CD62L, CCR7, CD28, and CD127, and positive for granzyme B and perforin. In some specific modalities, CD4+ T cells are further classified into subpopulations.For example, CD4+ helper T cells can be classified into naïve, central memory, and effector cells by identifying cell populations that have cell surface antigens. b. Immune cells derived from stem cells In some modalities, immune cells can be derived from embryonic stem cells (ES) or induced pluripotent stem cells (IPS). Suitable HSCs, mesenchymal stem cells, iPS cells, and other stem cell types can be cultured immortal cell lines or isolated directly from a patient. Several methods for isolating, growing, and / or culturing stem cells are known in the technique and can be used to implement the present description. In some modalities, the immune cell is an induced pluripotent stem cell (PSC) derived from a reprogrammed T cell. In some modalities, the source material may be an induced pluripotent stem cell (PSC) derived from a T cell or a non-T cell. The source material may be an embryonic stem cell. The source material may be a B cell, or any other cell type, including peripheral blood mononuclear cell isolates, hematopoietic progenitor cells, hematopoietic stem cells, mesenchymal stem cells, adipose stem cells, or any other type of somatic cell. c. Genetic modification of isolated cells Immune cells, such as T cells, can be genetically modified after isolation using known methods, or immune cells can be activated and expanded (or differentiated in the case of progenitor cells) in vitro before being genetically modified. In some modalities, isolated immune cells are genetically modified to reduce or eliminate the expression of endogenous TCRa and / or CD52. In some modalities, cells are genetically modified using gene-editing technology (e.g., CRISPR / Cas9, a zinc finger nuclease (ZFN), a TALEN, a MegaTAL, a meganuclease) to reduce or eliminate the expression of endogenous proteins (e.g., TCRa and / or CD52).In another modality, immune cells, such as T cells, are genetically modified with the chimeric antigen receptors described in the present description (e.g., transduced with a viral vector comprising one or more nucleotide sequences encoding a CAR) and then activated and / or expanded in vitro. PCT application PCT / US15 / 14520 describes certain methods for manufacturing the constructs and manipulated immune cells described herein, the contents of which are hereby incorporated by reference in their entirety. It will be noted that PBMCs may also include other cytotoxic lymphocytes such as NK cells or NKT cells. An expression vector carrying the coding sequence of a receptor The chimeric MA / a / ¿U¿l / Ul dUUO as described herein can be introduced into a population of human donor T cells, NK cells, or NKT cells. Successfully transduced T cells carrying the expression vector can be sorted using flow cytometry to isolate CD3-positive T cells and then further propagated to increase the number of these CAR-expressing T cells, in addition to cell activation using anti-CD3 and IL-2 antibodies or other methods known in the art as described elsewhere herein. Standard procedures are used for the cryopreservation of CAR-expressing T cells for storage and / or preparation for use in a human subject. In one modality, the transduction, culture, and / or in vitro expansion of T cells are performed in the absence of non-human animal-derived products such as fetal calf serum and fetal bovine serum. For polynucleotide cloning, the vector can be introduced into a host cell (an isolated host cell) to allow replication of the vector itself and thus amplify the copies of the polynucleotide it contains. Cloning vectors may contain sequence components that generally include, but are not limited to, an origin of replication, promoter sequences, transcription start sequences, enhancer sequences, and selection markers. These elements can be selected as appropriate by a person skilled in the technique. For example, the origin of replication can be selected to promote autonomous replication of the vector in the host cell. In certain embodiments, this description provides isolated host cells containing the vector provided herein. These vector-containing host cells may be useful for the expression or cloning of the polynucleotide contained within the vector. Suitable host cells may include, but are not limited to, prokaryotic cells, fungal cells, yeast cells, or higher eukaryotic cells such as mammalian cells, particularly human cells. The vector can be introduced into the host cell using any suitable method known in the art, including, but not limited to, DEAedextran-mediated delivery, calcium phosphate precipitation, cationic lipid-mediated delivery, liposome-mediated transfection, electroporation, microprojectile bombardment, receptor-mediated gene delivery, and delivery via polylysine, histone, chitosan, and peptides. Standard methods for transfecting and transforming cells to express a vector of interest are well known in the art. In a further modality, a mixture of different expression vectors can be used to genetically modify a donor population of immune effector cells, where each vector encodes a different CAR as described herein.The resulting transduced immune effector cells form a mixed population of manipulated cells, in which a proportion of the manipulated cells express more than one different CAR. In one modality, the description provides a method for storing genetically engineered cells that express CARs or TCRs. This involves cryopreservation of immune cells so that they remain viable upon thawing. A fraction of the CAR-expressing immune cells can be cryopreserved using methods known in the art to provide a permanent source of such cells for the future treatment of patients with malignancies. When needed, the cryopreserved transformed immune cells can be thawed, cultured, and expanded to obtain more of these cells. In some modalities, cells are first prepared by harvesting them from their culture medium, then washing and concentrating them in a suitable medium and container system for administration (a pharmaceutically acceptable carrier) in an amount effective for treatment. The suitable infusion medium can be any isotonic medium formulation, typically normal saline, Normosol™ R (Abbott) or Plasma-Lyte™ A (Baxter), but 5% dextrose in water or Ringer's lactate can also be used. The infusion medium can be supplemented with human serum albumin. 2Qd. Allogeneic CAR-T cells (ALLOCAR T™) The process for manufacturing allogeneic CAR-T cell therapy, or AlloCARs™, involves collecting healthy, selected, screened, and evaluated T cells from healthy donors. These T cells are then engineered to express CARs, which recognize specific cell surface proteins expressed in hematologic or solid tumors. The allogeneic T cells undergo gene editing to reduce the risk of graft-versus-host disease (GvHD) and to prevent allogeneic rejection. A T-cell receptor gene (e.g., TCRa, TCRP) is deactivated to prevent GvHD. The CD52 gene can be deleted to make the CAR-T cell product resistant to anti-CD52 antibody treatment. Therefore, anti-CD52 antibody treatment can be used to suppress the host's immune system and allow the CAR-T cell to remain engrafted to achieve its full therapeutic effect.The manipulated T cells then undergo a purification stage and are ultimately cryopreserved in vials for delivery to patients. and. Autologous CAR-T cells (AUTOCAR T™) Autologous chimeric antigen receptor (CAR) T-cell therapy involves collecting the patient's own cells (e.g., white blood cells, which include T cells) and genetically manipulating the T cells to express CARs that recognize the target expressed on the cell surface of one or more cancer cells and destroy the cancer cells. The manipulated cells are then cryopreserved and subsequently administered to the patient. 5. Pharmaceutical compositions and therapy In some modalities, cells are first prepared by harvesting them from their culture medium, then washing and concentrating them in a suitable medium and container system for administration (a pharmaceutically acceptable carrier) in an amount effective for treatment. The appropriate infusion medium may be any isotonic medium formulation, typically normal saline, Normosol™ R (Abbott), or Plasma-Lyte™ A (Baxter), but 5% dextrose in water or Ringer's lactate may also be used. The infusion medium may be supplemented with human serum albumin. In these formulations, the desired number of treatment cells in the composition is generally at least 2 cells (e.g., at least 1 CD8+ core memory T cell or stem cell and at least 1 CD4+ helper T cell subset; or two or more CD8+ core memory T cells or stem cells; or two or more CD4+ helper T cell subsets) or is more typically greater than 10² cells, and up to and including 10⁶, up to 20, and including 10⁷, 10⁸, or 10⁹ cells, and may be more than 10¹⁰ cells. The number of cells will depend on the intended use of the composition and the type of cells included. The desired cell density is typically greater than 10⁶ cells / ml and is generally greater than 10⁷ cells / ml, usually 10⁸ cells / ml or more. The clinically relevant number of immune cells can be distributed in multiple infusions that cumulatively equal or exceed 105, 106, 107, 108, 109, 1010, 1011 or 1012 cells.In some aspects of this description, particularly because all infused cells will be redirected to a specific target antigen, a smaller number of cells may be administered, in the range of approximately 10⁵ / kg or approximately 10⁶ / kg (10⁶–10¹¹ per patient). CAR T-cell therapies may be administered multiple times in doses within these ranges. The cells may be autologous, allogeneic, or heterologous to the patient undergoing the therapy. The CAR-expressing cell populations described herein may be administered alone or as a pharmaceutical composition in combination with diluents and / or other components such as IL-2 or other cytokines or cell populations. The pharmaceutical compositions described herein may comprise a CAR- or TCR-expressing cell population, such as T cells, as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents, or excipients. Such compositions may comprise buffers such as neutral buffered saline, phosphate-buffered saline, and the like; carbohydrates such as glucose, mannose, sucrose, or dextrans, mannitol; proteins; Polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives. The compositions described herein are preferably formulated for intravenous administration. Pharmaceutical compositions (solutions, suspensions or the like) may include one or more of the following: sterile diluents such as water for injection, saline solution, preferably physiological saline solution, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono- or diglycerides that can serve as a solvent or suspension medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; Buffers such as acetates, citrates, or phosphates and tonicity-adjusting agents such as sodium chloride or dextrose are used. The parenteral preparation may be contained in ampoules, disposable syringes, or multidose vials made of glass or plastic. An injectable pharmaceutical composition is preferably sterile. 206. Treatment methods The description includes methods for treating or preventing a disease (e.g., cancer) in a patient, comprising administering to a patient in need an effective amount of at least one CAR or immune cell comprising a CAR described herein. Methods for treating diseases or disorders, including cancer, are provided. In some modalities, the description refers to creating a T-cell-mediated immune response in a subject, comprising administering to the subject an effective quantity of the manipulated immune cells of this application. In some modalities, the T-cell-mediated immune response is directed against a target cell or cells. In some modalities, the manipulated immune cell comprises a chimeric antigen receptor (CAR). In some modalities, the target cell is a tumor cell. In some aspects, the description comprises a method for treating or preventing a malignant neoplasm, said method comprising administering to a subject in need an effective quantity of at least one isolated antigen-binding domain described herein.In some 35 aspects, the description comprises a method for treating or preventing a malignant neoplasm, said method comprising administering to a subject in need an effective amount of al. MA / a / ZUZl / Ul dUUO less than one immune cell, wherein the immune cell comprises at least one chimeric antigen receptor, T-cell receptor, and / or isolated antigen-binding domain as described herein. The CAR containing immune cells of the description can be used to treat neoplasms involving atypical biomarker expression. In some modalities, the immune cells containing CARs of the description can be used to treat small cell lung cancer, melanoma, low-grade gliomas, glioblastoma, medullary thyroid cancer, carcinoids, neuroendocrine tumors scattered in the pancreas, bladder, and prostate, testicular cancer, and lung adenocarcinomas with neuroendocrine features. In illustrative modalities, the immune cells containing CARs, for example, the CAR-T cells of the description, are used to treat small cell lung cancer. Methods for reducing the size of a tumor in a subject are also provided, comprising administering to the subject a manipulated cell of the present description, wherein the cell comprises a chimeric antigen receptor comprising an antigen-binding domain and binds to an antigen in the tumor. In some modalities, the subject has a solid tumor or a hematologic malignancy, such as lymphoma or leukemia. In some modalities, the manipulated cell is administered to a tumor bed. In some modalities, the cancer is present in the subject's bone marrow. In some modalities, the manipulated cells are autologous immune cells, for example, autologous T cells. In some modalities, the manipulated cells are allogeneic immune cells, for example, allogeneic T cells. In some modalities, the manipulated cells are heterologous immune cells, for example, heterologous T cells. In some modalities, the manipulated cells of this application are transfected or transduced in vivo. In other modalities, the manipulated cells are transfected or transduced ex vivo. As used herein, the term "in vitro cell" refers to any cell that is cultured ex vivo. A therapeutically effective amount, effective dose, or therapeutically effective amount of a therapeutic agent, for example, engineered CAR cells, is any quantity that, when used alone or in combination with another therapeutic agent, protects a subject against the onset of a disease or promotes disease regression, as evidenced by a decrease in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods, or a prevention of deterioration or disability due to the pathological condition. The ability of a therapeutic agent to promote disease regression can be assessed using a variety of methods known to the skilled clinician, such as in human subjects during clinical trials, in animal model systems that predict efficacy in humans, or by assaying the agent's activity in in vitro assays. The terms patient and subject are used interchangeably and include human subjects and non-human animals, as well as those with formally diagnosed disorders, those without formally recognized disorders, those receiving medical care, those at risk of developing disorders, etc. The terms "treat" and "treatment" include therapeutic treatments, prophylactic treatments, and applications that reduce the risk of an individual developing a disorder or other risk factor. Treatment does not require a complete cure for a disorder and includes modalities that reduce symptoms or underlying risk factors. The term "prevent" does not require the complete elimination of the possibility of an event. Rather, it indicates that the likelihood of the event occurring has been reduced in the presence of the compound or method. The desired number of treatment cells in the composition is generally at least 2 cells (e.g., at least 1 CD8+ core memory T cell and at least 1 subset of CD4+ helper T cells) or more typically greater than 10² cells, and up to 10⁶, including 10⁸ or 10⁹ cells, and may be more than 10¹⁰ cells. The number of cells will depend on the intended use of the composition and the type of cells included. The desired cell density is typically greater than 10⁶ cells / ml and generally greater than 10⁷ cells / ml, usually 10⁸ cells / ml or more. The clinically relevant number of immune cells can be distributed in multiple infusions that cumulatively equal or exceed 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, or 10¹² cells.In some aspects of this description, particularly because all infused cells will be redirected to a specific target antigen, a smaller number of cells may be administered, in the range of 10⁶ / kilogram (10⁶–10¹¹ per patient). CAR T-cell therapies can be administered multiple times in doses within these ranges. The cells may be autologous, allogeneic, or heterologous to the patient undergoing the therapy. In some modalities, the therapeutically effective amount of CAR-T cells is approximately 1 x 10⁵ cells / kg, approximately 2 x 10⁵ cells / kg, approximately 3 x 10⁵ cells / kg, approximately 4 x 10⁵ cells / kg, approximately 5 x 10⁵ cells / kg, approximately 6 x 10⁵ cells / kg, approximately 7 x 10⁵ cells / kg, approximately 8 x 10⁵ cells / kg, approximately 9 x 10⁵ cells / kg, 2 x 10⁶ cells / kg, approximately 3 x 10⁶ cells / kg, approximately 4 x 10⁶ cells / kg, approximately 5 x 10⁶ cells / kg, approximately 6 x 10⁶ cells / kg, approximately 7 x 10⁶ cells / kg, approximately 8 x 10⁶ cells / kg, approximately 9 x 106cells / kg, approximately 1 X 107cells / kg, approximately 2 X 107cells / kg, approximately 3 107cells / kg or approximately 9 X 107cells / kg. In some modalities, the target doses for CAR+ / CAR-T+ / TCR+ cells vary from 1 x 10⁶ cells / kg to, for example, 2 x 10⁶ cells / kg. It should be noted that doses above and below this range may be appropriate for certain individuals, and the healthcare provider can determine the appropriate dose levels as needed. Additionally, multiple cell doses may be provided as described. In some aspects, the description comprises a pharmaceutical composition comprising at least one antigen-binding domain as described herein and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further comprises an additional active agent. The CAR-expressing cell populations described herein may be administered alone or as a pharmaceutical composition in combination with diluents and / or other components such as IL-2 or other cytokines or cell populations. The pharmaceutical compositions described herein may comprise a CAR- or TCR-expressing cell population, such as T cells, as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents, or excipients. Such compositions may include buffers such as neutral buffered saline, phosphate-buffered saline, and the like; carbohydrates such as glucose, mannose, sucrose, or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives.The compositions described herein are preferably formulated for intravenous administration. Pharmaceutical compositions (solutions, suspensions, or the like) may include one or more of the following: sterile diluents such as water for injection, saline solution (preferably physiological saline), Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono- or diglycerides that may serve as a solvent or suspension medium, polyethylene glycols, glycerin, propylene glycol, or other solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates, or phosphates; and tonicity-adjusting agents such as sodium chloride or dextrose. The parenteral preparation may be contained in ampoules, disposable syringes, or multidose vials made of glass or plastic. An injectable pharmaceutical composition is preferably sterile. In some embodiments, following administration to a patient, engineered immune cells expressing any of the antigen-specific CARs described herein can reduce, destroy, or lyse the patient's cells expressing endogenous antigens. In one embodiment, the percentage reduction or lysis of endogenous antigen-expressing cells or cells of a cell line expressing an antigen by engineered immune cells expressing any of the antigen-specific CARs described herein is at least approximately or greater than 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.In one modality, a percentage reduction or lysis of endogenous cells expressing an antigen or cells of a cell line expressing an antigen by manipulated immune cells expressing antigen-specific CARs is approximately 5% to approximately 95%, approximately 10% to approximately 95%, approximately 10% to approximately 90%, approximately 10% to approximately 80%, approximately 10% to approximately 70%, approximately 10% to approximately 60%, approximately 10% to approximately 50%, approximately 10% to approximately 40%, approximately 20% to approximately 90%, approximately 20% to approximately 80%, approximately 20% to approximately 70%, approximately 20% to approximately 60%, approximately 20% to approximately 50%, approximately 25% to approximately 75%, or approximately 25% to approximately 60%.In one modality, the cells that express endogenous antigens are bone marrow cells that express endogenous antigens. In one modality, the percentage reduction or lysis of target cells, for example, a cell line expressing an antigen, by manipulated immune cells expressing on their cell surface membrane an antigen-specific CAR of the description, can be measured using the assay described herein. The methods may further include administering one or more chemotherapeutic agents. In certain modalities, the chemotherapeutic agent is a lympho-depleting (preconditioning) chemotherapeutic agent. For example, methods for conditioning a patient who requires T-cell therapy include administering to the patient specific beneficial doses of cyclophosphamide (between 200 mg / m² / day and 2,000 mg / m² / day, approximately 100 mg / m² / day and approximately 2,000 mg / m² / day).for example, approximately 100 mg / m2 / day, approximately 200 mg / m2 / day, approximately 300 mg / m2 / day, approximately 400 mg / m2 / day, approximately 500 mg / m2 / day, approximately 600 mg / m2 / day, approximately 700 mg / m2 / day, approximately 800 mg / m2 / day, approximately 900 mg / m2 / day, approximately 1,000 mg / m2 / day, approximately 1,500 mg / m2 / day or approximately 2,000 mg / m2 / day) and specific doses of fludarabine (between 20 mg / m2 / day and 900 mg / m2 / day, between approximately 10 mg / m2 / day and approximately 900 mg / m2 / day;for example, approximately 10 mg / m2 / day, approximately 20 mg / m2 / day, approximately 30 mg / m2 / day, approximately 40 mg / m2 / day, approximately 40 mg / m2 / day, approximately 50 mg / m2 / day, approximately 60 mg / m2 / day, approximately 70 mg / m2 / day, approximately 80 mg / m2 / day, approximately 90 mg / m2 / day, approximately 100 mg / m2 / day, approximately 500 mg / m2 / day, or approximately 900 mg / m2 / day). A preferred dosing regimen involves treating a patient comprising administering to the patient daily approximately 300 mg / m2 / day of cyclophosphamide and approximately 30 mg / m2 / day of fludarabine for three days prior to administering to the patient a therapeutically effective amount of manipulated T cells. In some modalities, lymphatic depletion also includes the administration of an antibody against CD52. In some modalities, the CD52 antibody is administered at a dose of approximately 13 mg / day IV. In other modalities, the antigen-binding domain, the transduced (or otherwise manipulated) cells, and the chemotherapeutic agent are each administered in an amount effective to treat the disease or condition in the subject. In certain formulations, the CAR-expressing immune effector cell compositions described herein may be administered in conjunction with any variety of chemotherapeutic agents, which may be administered in any order. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN™); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; azirlidines such as benzodopa, carboquone, meturedopa, and uredopa; and ethyleneimiles and methylamylamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine. nitrogen mustards such as chlorambucil, chlornaphazine, colofosfamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, fenesterin, prednimustine, trofosfamide, uracil mustard;nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomycins, actinomycin, autramycin, azaserine, bleomycins, cactinomycin, calicheamycin, carabicin, carminomycin, carzinophylline, cromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxino-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, chelamicin, rhodorubicin, streptonigrine, streptozocin, tubercidine, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as; MA / a / ZUZl / Ul dUUO such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxyfluridine, enocitabine, floxuridine, 5-FU; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid booster such as frolinic acid; aceglatone; aldophosfamide glucoside; aminolevulinic acid; amsacrine; bestrabucil; bisanthrene; edatrataxate; defofamine; demecolcine; diaziquone; elformitin; elliptinium acetate; ethoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; fenamet; pirarubicin; podophylinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; sizofiran; spirogermanium; tenuazonic acid;triazicuone; 2,2',2-trichlorotiet¡lam¡na; urethane; vindesine; dacarbazine; manomustine; mitobronitol; mitolactol; pipobroman; gacytosin; arabinoside (Ara-C); cyclophosphamide; thiotepa; taxoides, for example, paclitaxel (TAXOL™, Bristol-Myers Squibb) and doxetaxel (TAXOTERE®, RhonePoulenc Rorer); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbina; novantrona; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RF S2000; difluoromethylomitin (DMFO); retinoic acid derivatives such as Targretin™ (bexarotene), Panretin™ (alitretinoin); ONTAK™ (denyleukin diftitox); esperamycins; capecitabine;and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing. Also included in this definition are antihormonal agents that act to regulate or inhibit hormonal action in tumors, such as antiestrogens, including, for example, tamoxifen, raloxifene, 4(5)-imidazole aromatase inhibitors, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston); and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing. Combinations of chemotherapeutic agents are also administered when appropriate, including, but not limited to, CHOP, i.e., cyclophosphamide (Cytoxan®), doxorubicin (hydroxydoxorubicin), vincristine (Oncovin®), and prednisone. In some modalities, the chemotherapeutic agent is administered at the same time as, or within one week of, the administration of the manipulated cell, polypeptide, or nucleic acid. In other modalities, the chemotherapeutic agent is administered 1 to 4 weeks, or 1 week to 1 month, 1 week to 2 months, 1 week to 3 months, 1 week to 6 months, 1 week to 9 months, or 1 week to 12 months after the administration of the manipulated cell, polypeptide, or nucleic acid. In still other modalities, the chemotherapeutic agent is administered at least 1 month before the administration of the cell, polypeptide, or nucleic acid. In some modalities, the methods also involve administering two or more chemotherapeutic agents. A variety of additional therapeutic agents may be used in conjunction with the compositions described herein. For example, potentially useful additional therapeutic agents include PD-1 inhibitors such as nivolumab (Opdivo®), pembrolizumab (Keytruda®), pidilizumab, and atezolizumab. The additional therapeutic agents suitable for use in combination with the description include, but are not limited to, ibrutinib (Imbruvica®), ofatumumab (Arzerra®, rituximab (Rituxan®), bevacizumab (Avastin®), trastuzumab (Herceptin®), trastuzumab emtansine (KADCYLA®), imatinib (Gleevec®), cetuximab (Erbitux®), panitumumab (Vectibix®), catumaxomab, ibritumomab, ofatumumab, tositumomab, brentuximab, alemtuzumab, gemtuzumab, erlotinib, gefitinib, vandetanib, afatinib, lapatinib, neratinib, axitinib, masitinib, pazopanib, sunitinib, sorafenib, toceranib, lestaurtinib, axitinib, cediranib, lenvatinib, nintedanib, pazopanib, regorafenib, semaxanib, sorafenib, sunitinib, tivozanib, toceranib, vandetanib, entrectinib, cabozantinib, imatinib, dasatinib, nilotinib, ponatinib, radotinib, bosutinib, lestaurtinib, ruxolitinib, pacritinib, cobimetinib, selumetinib, trametinib, binimetinib, alectinib, ceritinib, crizotinib, aflibercept, adipotida, denileucina diftitox,mTOR inhibitors such as everolimus and temsirolimus, hedgehog inhibitors such as sonidegib and vismodegib, CDK inhibitors such as the CDK inhibitor (palbociclib). In some formulations, the CAR-containing immune cell composition may be administered with a therapeutic regimen to prevent cytokine release syndrome (CRS) or neurotoxicity. The therapeutic regimen to prevent CRS or neurotoxicity may include lenzilumab, tocilizumab, atrial natriuretic peptide (ANP), anakinra, or NOS inhibitors (e.g., L-NIL or 1400W). In additional formulations, the CAR-containing immune cell composition may be administered with an anti-inflammatory agent.Anti-inflammatory agents or drugs include, but are not limited to, steroids and glucocorticoids (including betamethasone, budesonide, dexamethasone, hydrocortisone acetate, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone), and nonsteroidal anti-inflammatory drugs (NSAIDs) including aspirin, ibuprofen, naproxen, methotrexate, sulfasalazine, leflunomide, anti-TNF drugs, cyclophosphamide, and mycophenolate. Illustrative NSAIDs include ibuprofen, naproxen, naproxen sodium, COX-2 inhibitors, and sialilates. Illustrative analgesics include acetaminophen, oxycodone, tramadol, and proproxyphene hydrochloride. Illustrative glucocorticoids include cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, or prednisone. Illustrative biological response modifiers include molecules directed against cell surface markers (e.g., CD4, CD5, etc.).), cytokine inhibitors, such as TNF antagonists (e.g., etanercept (ENBREL®), adalimumab (HUMIRA®), and infliximab (REMICADE®), chemokine inhibitors, and adhesion molecule inhibitors. The 5 biological response modifiers include monoclonal antibodies as well as recombinant forms of molecules. Illustrative DMARDs include azathioprine, cyclophosphamide, cyclosporine, methotrexate, penicillamine, leflunomide, sulfasalazine, hydroxychloroquine, Gold (oral (auranofin) and intramuscular), and minocycline. In certain formulations, the compositions described herein are administered together with a cytokine. Examples of cytokines include lymphokines, monokines, and traditional polypeptide hormones. Cytokines include growth hormones such as human growth hormone, human N-methionyl growth hormone, and bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones such as follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), and luteinizing hormone (LH); liver growth factor (HGF); fibroblast growth factor (FGF); prolactin; placental lactogen; Müller inhibitory substance; mouse gonadotropin-associated peptide; inhibin; activin; vascular endothelial growth factor; integrin; and thrombopoietin (TPO). nerve growth factors (NGF) such as NGF-beta;platelet growth factor; transforming growth factors (TGFs) such as TGF-alpha and TGF-beta; insulin-like growth factor I and II; erythropoietin (EPO); osteoinductive factors; interferons such as interferon-alpha, beta, and gamma; colony-stimulating factors (CSFs) such as macrophage CSF (M-CSF); granulocyte-macrophage CSF (GM-CSF); and granulocyte-macrophage CSF (GCSF); interleukins (ILs) such as IL-1, IL-1 alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12; IL-15, IL-21, a tumor necrosis factor such as TNF-alpha or TNF-beta; and other polypeptide factors including LIF and kit ligand (KL). As used in this description, the term cytokine includes proteins from natural sources or recombinant cell culture and biologically active equivalents of native sequence cytokines. 307. Classification and exhaustion methods In some embodiments, methods are provided for the in vitro sorting of an immune cell population, wherein a subset of the immune cell population comprises engineered immune cells expressing antigen-specific CARs comprising epitopes specific to monoclonal antibodies (e.g., illustrative 35-mimotope sequences). The method comprises contacting the immune cell population with a monoclonal antibody specific to the epitopes and selecting the immune cells that bind to the monoclonal antibody to obtain a cell population enriched in engineered immune cells expressing an antigen-specific CAR. In some embodiments, the epitope-specific monoclonal antibody is optionally conjugated to a fluorophore.In this modality, the step of selecting the cells that bind to the monoclonal antibody can be performed by fluorescence-activated cell sorting (FACS). In some formulations, the monoclonal antibody specific for the epitope is optionally conjugated to a magnetic particle. In this formulation, the step of selecting the cells that bind to the monoclonal antibody can be performed using magnetically activated cell sorting (MACS). In some embodiments, the mAb used in the method to sort immune cells expressing the CAR is selected from alemtuzumab, ibritumomab tiuxetan, muromonab-CD3, tositumomab, abciximab, basiliximab, brentuximab, vedotin, cetuximab, infliximab, rivacimab, bevacizumab, certolizumab pegol, daclizumab, eculizumab, efalizumab, gemtuzumab, natalizumab, omalizumab, palivizumab, ranibizumab, tocilizumab, trastuzumab, vedolizumab, adalimumab, belimumab, canakinumab, denosumab, golimumab, iatumumab, panitumumab, panitumumab, QBEND-10 y / o ustekinumab. In some embodiments, said mAb is rituximab. In another embodiment, said mAb is QBEND-10. In another modality, the mAb binds to TCRa or TCRp. In some modalities, the CAR-expressing immune cell population obtained using the method for classifying CAR-expressing immune cells described above comprises at least 70%, 75%, 80%, 85%, 90%, or 95% of CAR-expressing immune cells. In some modalities, the CAR-expressing immune cell population obtained using the method for classifying CAR-expressing immune cells comprises at least 85% of CAR-expressing immune cells. In some modalities, the CAR-expressing immune cell population obtained using the previously described method for in vitro sorting of CAR-expressing immune cells exhibits increased in vitro cytotoxic activity compared to the initial (unsorted) cell population. In some modalities, this in vitro cytotoxic activity increases by 10%, 20%, 30%, or 50%. In some modalities, the immune cells are T cells. In some formulations, the mAbs are pre-attached to a support or surface. Non-limiting examples of solid supports may include a bead, an agarose bead, a magnetic bead, a plastic rimmed plate, a glass rimmed plate, a ceramic rimmed plate, a column, or a cell culture bag. CAR-expressing immune cells to be administered to the recipient can be enriched in vitro from the source population. Methods for expanding source populations may include selecting cells that express an antigen such as the CD34 antigen, using combinations of density centrifugation, immunomagnetic bead purification, affinity chromatography, and fluorescence-activated cell sorting. Flow cytometry can be used to quantify specific cell types within a cell population. In general, flow cytometry is a method for quantifying structural components or characteristics of cells, primarily by optical means. Since different cell types can be distinguished by quantifying structural characteristics, flow cytometry and cell sorting can be used to count and classify cells of different phenotypes in a mixture. A flow cytometry analysis involves two main steps: 1) labeling selected cell types with one or more labeled markers, and 2) determining the number of labeled cells relative to the total number of cells in the population. In some methods, the cell type labeling process involves attaching labeled antibodies to markers expressed on the specific cell type. Antibodies can be labeled directly with a fluorescent compound or indirectly by using, for example, a second fluorescently labeled antibody that recognizes the first antibody. In some modalities, the method used to classify CAR-expressing T cells is magnetically activated cell sorting (MACS). MACS is a method for separating various cell populations based on their surface antigens (CD molecules) using superparamagnetic nanoparticles and columns. MACS can be used to obtain a pure cell population. Cells in a single-cell suspension can be magnetically labeled with microbeads. The sample is applied to a column composed of ferromagnetic spheres, which are coated with a cell-nondamaging material, allowing for rapid and gentle cell separation. Unlabeled cells pass through, while magnetically labeled cells are retained within the column. The throughflow can be collected as the unlabeled cell fraction.After a washing stage, the column is removed from the separator and the magnetically labeled cells are eluted from the column. The detailed protocol for the purification of a specific cell population, such as T cells, can be found in Basu S et al. (2010). (Basu S, Campbell HM, Dittel BN, Ray A. Purification of specific cell population by fluorescence activated cell sorting (FACS). J Vis Exp. (41): 1546). In some respects, the present description provides a method for depleting antigen-specific CAR-expressing immune cells by in vivo exhaustion. In vivo exhaustion may involve administering a treatment (e.g., a molecule that binds to an epitope on the CAR) to a mammalian organism with the aim of halting the proliferation of CAR-expressing immune cells by inhibition or elimination. One aspect of the description relates to a method for in vivo exhausting a genetically engineered immune cell expressing a CAR comprising a mAb-specific epitope, by contacting the engineered immune cell or the CAR-expressing immune cell with at least one epitope-specific mAb. Another aspect of the description relates to a method for in vivo exhausting the CAR-expressing immune cell comprising a chimeric scFv (e.g., formed by the insertion of an mAb-specific epitope) by contacting the engineered immune cell with epitope-specific antibodies. In some embodiments, the immune cells are T cells and / or the antibodies are monoclonal. According to one embodiment, in vivo exhaustion of the manipulated immune cells is performed on manipulated immune cells that have been previously sorted using an in vitro method described herein. In this case, the same infused mAb may be used. In some embodiments, the epitope-specific mAb is the CD20 antigen and the epitope-specific mAb is rituximab. In some embodiments, the description refers to a method for in vivo exhaustion of a manipulated immune cell expressing a CAR comprising an epitope-specific mAb (CAR-expressing immune cell) in a patient, comprising contacting said CAR-expressing immune cell with at least one epitope-specific mAb. In some modalities, the step of contacting the engineered immune cell or the CAR-expressing immune cell with at least one epitope-specific mAb comprises infusing the patient with an epitope-specific mAb (e.g., rituximab). In some modalities, the amount of epitope-specific mAb administered to the patient is sufficient to eliminate at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the CAR-expressing immune cell in the patient. In some modalities, the step of contacting the engineered immune cell or the CAR-expressing immune cell with at least one epitope-specific mAb comprises infusing the patient with 375 mg / m² of rituximab, once or multiple times. In some modalities, the mAb (e.g., rituximab) is administered once a week. In some modalities, when immune cells expressing a CAR containing a mAb-specific epitope (CAR-expressing immune cells) are depleted in a complement-dependent cytotoxicity (CDC) assay using an epitope-specific mAb, the number of viable CAR-expressing immune cells decreases. In some modalities, the number of viable CAR-expressing immune cells decreases by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In some modalities, the mAb-specific epitope is a CD20 epitope or mimotope, and / or the epitope-specific mAb is rituximab. In certain modalities, in vivo depletion of CAR-manipulated immune cells is achieved by infusing bispecific antibodies. By definition, a bispecific monoclonal antibody (BsAb) is an artificial protein composed of fragments of two different monoclonal antibodies and, consequently, binds to two different types of antigen. These BsAbs and their use in immunotherapy have been reviewed in Muller D and Kontermann RE (2010) Bispecific Antibodies for Cancer Immunotherapy, BioDrugs 24 (2): 89-98. According to another particular modality, the infused bispecific mAb can bind both to the mAb-specific epitope found on manipulated immune cells expressing the chimeric scFv and to a surface antigen on an effector, cytotoxic cell (e.g., immune cells such as lymphocytes, macrophages, dendritic cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTLs)). By doing so, the depletion of manipulated immune cells triggered by the BsAb can occur through antibody-dependent cell-mediated cytotoxicity (ADCC). (Deo YM, Sundarapandiyan K, Keler T, Wallace PK, and Graziano RF, (2000), Journal of Immunology, 165 (10):5954-5961). In some formulations, a cytotoxic drug is coupled to epitope-specific mAbs that can be used to deplete CAR-expressing immune cells. By combining the specific recognition capabilities of monoclonal antibodies with the cancer-destroying ability of cytotoxic drugs, the antibody-drug conjugate (ADC) allows for sensitive discrimination between healthy and diseased tissue compared to using the drug alone. Market approvals have been received for several ADCs; The technology for manufacturing them - particularly linkers - is described in (Payne, G. (2003) Cancer Cell 3:207-212; Trail et al., (2003) Cancer Immunol. Immunother.52:328-337; Syrigos and Epenetos (1999) Anticancer Research 19:605-614; Niculescu-Duvaz and Springer (1997) Adv. Drug Del. Rev. 26:151-172; U.S. patent no. 4,975,278). MA / a / ZUZl / Ul dUUO In some formulations, the epitope-specific mAb to be infused is pre-conjugated with a molecule that can promote complement-dependent cytotoxicity (CDC). The complement system thus assists or complements the ability of antibodies to eliminate pathogens from the body. When stimulated, it triggers an activation cascade, including massive amplification of the response and activation of the membrane attack complex, which destroys cells. Different molecules can be used to conjugate the mAb, such as glycans [Courtois, A., Gac-Breton, S., Berthou, C., Guezennec, J., Bordron, A., & Boisset, C. (2012). Complement-dependent cytotoxicity activity of therapeutic antibody fragments can be acquired by immunogenic glycan coupling. Electronic Journal of Biotechnology 10 ISSN: 0717-3458; http: / / www.ejbiotechnology.info DOI: 10.2225 / voll5-issue5]. 8. Manufacturing kits and items This description provides kits comprising any of the cultured immune cells or manipulated immune cells described herein, and 15 pharmaceutical compositions thereof. In some illustrative embodiments, a kit of the description comprises allogeneic CAR-containing T cells for administration to a subject. This application also provides for manufactured items comprising any of the therapeutic compositions or kits described herein. Examples of a manufactured item include vials (e.g., sealed vials). This description further provides kits comprising one or more containers comprising a potassium solution at a desired concentration and, optionally, one or more containers comprising a solution comprising one or more cytokines, such as IL-7 or IL-15. EXAMPLES As the following examples illustrate, combinations of conditions and concentrations of cytokines and metabolic modulators in T-cell expansion media can lead to increased potency of genetically modified allogeneic cell therapy products. For example, desirable allogeneic CAR-T cell phenotypes can be achieved through the use of cell proliferation stimulants, such as IL-7 and IL-15, and increased extracellular potassium. 1. Illustrative protocol for the production of CAR-T cells As described herein, CAR-T cells can be produced according to 35 methods known in the art. An illustrative standard method is described herein. To generate CAR-T cells, PBMCs can be purified from bleb layer samples from healthy volunteers using Ficoll gradient density medium (Ficoll Paque PLUS / GE Healthcare Life Sciences). T cells can be purified from PBMCs using a commercially available T cell isolation kit (MiItenyi Biotec, Cat# 130-096-535). Alternatively, primary human T cells can be purified directly from Leuko Paks (StemCell Technologies). To produce CAR-encoding lentiviruses, HEK-293T cells can be seeded at 0.4 million cells per ml in 2 ml of DMEM (Gibco) supplemented with 10% FBS (Hyclone or JR Scientific) per well of a 6-well plate on day 0. On day 1, the lentivirus can be prepared by mixing lentiviral packaging vectors: 1.5 µg of psPAX2, 0.5 µg of pMD2G, and 0.5 µg of the appropriate CAR transfer vector in 250 µl of Opti-MEM (Gibco) per well of the 6-well plate (“DNA mix”). 10 µl of Lipofectamine 2000 (Invitrogen) can be incubated in 250 µl of Opti-MEM at room temperature for 5 minutes and then added to the DNA mix. The mixture can be incubated at room temperature for 20 minutes and the total volume of 500 ul was slowly added to the sides of the wells containing HEK-293T. Purified T cells can be activated in a suitable medium. On day 2, the medium in each well of the 6-well plate can be replaced with 2 mL per well of T cell transduction medium, i.e., X-Vivo-15 supplemented with 10% FBS. On day 3, the T cells can be resuspended at 0.5 million cells per mL in 1 mL of T cell transduction medium per well of a Grex-24 plate (Wilson Wolff, cat# 80192M). The lentiviral supernatants from the HEK293T cells can be collected and passed through a 0.45-µm filter (EMD Millipore) to remove cellular debris and then added to the T cells along with 100 µL / mI of human IL-2. On day 5, 4.5 ml of T-cell expansion medium (e.g., comprising IL-7+IL-15 and with or without additional extracellular potassium) can be added to each well of a Grex-24 plate. On days 9 and 13, transduction efficiency can be determined by detecting the percentage of T cells that recognize the desired antigen (e.g., a recombinant antigen) using flow cytometry. The cells were expanded in larger flasks or G-Rex (Wilson Wolff) vessels as required using T-cell expansion media as described herein. On day 14, antigen-specific CAR-T cells can be cryopreserved, for example, by freezing the cells in a medium such as CryoStor® CS5, CryoStor® CS10, or CryoStor® CS2 (BioLife Solutions). The percentage of cells stained with recombinant antigen can be normalized between clones just before cryopreservation. 2. Complementation with IL-7+15 The data from the following examples demonstrate, for example, the use of a first stimulant and a second stimulant, for example, IL-7 (5,000 Ul / ml) and IL-15 (50 Ul / ml), optionally plus an increase in extracellular potassium (25 mM), can achieve a highly convenient phenotype of 5 allogeneic CAR-T cells, compared to classical IL-2-based processes. First, IL-7 and IL-15 complementation was evaluated in cell culture media in which potassium was maintained at the baseline level of 4 mM. As shown in Figure 1A, when compared to an IL-2-based manufacturing process (IL-2 -> IL-2” - IL-2 at 100 IU / ml throughout, fresh IL-2 added on day 5), an IL-7+IL-15-based process (either “IL-2 -> IL-7+15” - IL-2 at 100 IU / ml until day 2, on day 5, IL-7 at 5,000 IU / ml and IL-15 at 50 IU / ml were added, or “IL-7+15 -> IL-7+15” - IL-7 at 5,000 IU / ml and IL-15 at 50 IU / ml throughout with fresh IL-7 and IL-15 added on day 5) increased the abundance of Tscm (CD62L+CD45ROj CD19-specific CAR-T cells) in the final product. As shown in Figure 1B, Culture conditions with IL-7+15 also increased the release capabilities of 15 cytokines from CD19-specific CAR-T cells following exposure to target cells. 3. Potassium titration and supplementation studies To further enhance the potency of our allogeneic CAR-T cells, we investigated the combined benefits of IL-7 supplementation at the same concentrations as in previous experiments with extracellular nutrients known to modulate T cell metabolism. Extracellular potassium has been shown to be a suppressive mechanism by which the tumor microenvironment (TME) suppresses effector T cells in vivo. Conversely, increasing extracellular potassium during in vitro expansion of adoptive cell therapies (ACTs) leads to the preservation of T cells with a younger, less differentiated phenotype. As shown in Figure 2A, conditions of increased extracellular potassium (40 mM, unfilled bars) increase the abundance of Tscm cells of allogeneic anti-CD19 CAR-T cells in both IL-2- and IL-7+15-based processes compared to normal potassium concentrations (4 mM) (filled bars). In Figure 2B, however, it was unexpectedly found that conditions of increased extracellular potassium (40 mM, unfilled bars) negatively affect the expansion capacity of allogeneic CAR-T cells in both IL-2- and IL-7+15-based processes compared to normal potassium concentrations (4 mM, filled bars). To further enhance the benefits of high extracellular potassium during allogeneic CAR-T cell fabrication, the amount of extracellular potassium that is particularly effective for an IL-7+15-based allogeneic CAR-T cell process was adjusted. Extracellular potassium concentrations of 25 mM and 10 mM did not adversely affect human T cell expansion during allogeneic CAR-T cell fabrication (Figure 35). 3) . Furthermore, maximum Tscm cell preservation was achieved at extracellular potassium 25 mM (Figure 4). 4. Potency studies using combined supplementation with IL-7+15 and potassium A serial destruction assay involves repeatedly exposing CAR-T cells to their target, causing the CAR-T cells to proliferate and, in some cases, differentiate and exhaust. This assay was used to study the potency of allogeneic CAR-T cells expanded under different conditions. Having identified the effective extracellular potassium concentrations for manufacturing allogeneic CAR-T cells, we proceeded to evaluate whether combining IL-7+15 with a potassium concentration of 25 mM enhances the potency of our allogeneic CAR-T cells. Combining an IL-7+15-based process with 25 mM of extracellular potassium supplementation leads to maximum CAR-T cell potency compared to similar products manufactured with IL-2 at normal (4 mM) potassium concentrations (Figure 5). Furthermore, combining an IL-7+15-based process with 25 mM of extracellular potassium significantly improved potency compared to the IL-2-based process with potassium. 25 mM and even further improved the IL-7+15-based process with a baseline potassium level of 4 mM. In conclusion, the highly effective combination of cytokines (IL-7 + IL-15) and metabolic modulators (extracellular potassium) has been identified, which can maximize the potency of allogeneic CAR-T cells. Although the teachings described herein have been presented with reference to various applications, methods, kits, and compositions, it will be appreciated that several changes and modifications can be made without departing from the teachings of the present description and the invention claimed herein. The preceding examples are provided to further illustrate the teachings described herein and are not intended to limit the scope of the teachings presented herein. While the present teachings have been described in terms of these illustrative modalities, it will readily be understood by those skilled in the art that numerous variations and modifications of these illustrative modalities are possible without excessive experimentation. All such variations and modifications are within the scope of the present teachings. SEQ ID NO. SUMMARY TABLE SEQ ID NO. Description Sequence 5 1 Suicide polypeptide CPYSNPSLCSGGGGSELPTQGTFSNVSTNVSPAKPTTTACPYSNP SLCSGGGGSPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRG LDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRRRVCKCPRPVV 2 Suicide signal peptide MGTSLLCWMALCLLGADHADA 10 3 Suicide signal peptide and suicide sequence MGTSLLCWMALCLLGADHADACPYSNPSLCSGGGGSELPTQGTF SNVSTNVSPAKPTTTACPYSNPSLCSGGGGSPAPRPPTPAPTIAS QPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSL VITLYCNHRNRRRVCKCPRPVV 15 4 Rituximab Mimotope CPYSNPSLC 5 Palivizumab Epitope NSELLSLINDMPITNDQKKLMSNN 6 Cetuximab Mimotope 1 CQFDLSTRRLKC 20 7 Cetuximab Mimotope 2 CQYNLSSRALKC 8 Cetuximab Mimotope 3 CVWQRWQKSYVC 25 9 Cetuximab Mimotope 4 CMWDRFSRWYKC 10 Nivolumab Epitope 1 SFVLNWYRMSPSNQTDKLAAFPEDR 11 Nivolumab Epitope 2 SGTYLCGAISLAPKAQIKE 30 12 QBEND-10 Epitope 1 ELPTQGTFSNVSTNVS 25 Table 1 p. 20 QBEND-10 Epitope 2 ELPTQGTFSNVSTNVSPAKPTTA 13 Alemtuzuma b_Epitope GQNDTSQTSSPS 14 Hinge of FcyRIIIa GLAVSTISSFFPPGYQ ​​15 Hinge of CD8a Hinge of lgG1 EPKSPDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMIARTPEVTCV VDVSHEDPEVKFNWYVDGVEVHNACTKPREEQYNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP V LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK 17 Transmembrane na de CD8o IYIWAPLAGTCGVLLLSLVIT Transmembrane domain na CD28 FWVLVVVGGVLACYSLLVTVAFIIFWV 19 Domain zeta deCD3(1) LRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDP EMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGH DGLYQGLSTATKDTYDALHMQALPPR 26 Zeta domain of CD3 (2) LRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDP EMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGH DGLYQGLSTATKDTYDALHMQALPPR 20 4-1BB KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL 21 Nucleic acid sequence of 4- 1BB AAGCGCGGCAGGAAGAAGCTCCTCTACAI III IAAGCAGCCTTTTTTATGAGGCCCGTACAGACAACACAGGAGGAAGATGGCTGTAGC TGCAGATTTCCCGAGGAGGAGGAAGGTGGGTGCGAGCTG 22 Intracellular CD28 comprises the nucleic acid AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATAT GACTCC ACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCA CCACCTAGAGATTTCGCTGCCTATCGGAGC 23 Intracellular domain of CD3 zeta RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 24 Zeta nucleic acid sequence CD3 AGGGTGAAG lili CCAGATCTGCAGATGCACCAGCGTATCAGC AGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAG GGAAGAGTCAGGACCGGACCGGACCGGACCG GAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTC TCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCT GAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCAC

Claims

Having described the present invention, the following claims are considered novel and therefore claimed as priority: CLAIMS 1. A cell culture medium for T cell expansion comprising: a first cell proliferation stimulant and a second cell proliferation stimulant, each independently selected from the group consisting of IL-4, IL-7, IL-10, IL-12 and IL-15; and an extracellular modulator of cell metabolism which is extracellular potassium at a concentration of approximately 4 mM to approximately 40 mM, wherein the first stimulant and the second stimulant are present in a concentration ratio of approximately 1,000:1 to approximately 4:

1.

2. The cell culture medium according to claim 1, wherein the first cell proliferation stimulant is IL-7.

3. The cell culture medium according to claim 1 or 2, wherein the second cell proliferation stimulant is IL-15. 4.The cell culture medium of any of claims 1-3, wherein the first stimulant and the second stimulant are present in a concentration ratio of approximately 500:1 to approximately 10:1, approximately 250:1 to approximately 10:1, approximately 200:1 to approximately 10:1, approximately 150:1 to approximately 10:1, approximately 100:1 to approximately 10:1, approximately 500:1 to approximately 50:1, approximately 250:1 to approximately 50:1, approximately 200:1 to approximately 50:1, approximately 150:1 to approximately 50:1, approximately 100:1 to approximately 50:1, approximately 500:1 to approximately 75:1, approximately 250:1 to approximately 75:1, approximately 200:1 to approximately 75:1, approximately 150:1 to approximately 75:1, approximately 100:1 to about 75:1, about 10:1 to about 4:1, about 8:1 to about 4:1 or about 7:1 to about 6:

1.

5. The cell culture medium of any of claims 1-3, wherein the first stimulant and the second stimulant are present in a concentration ratio of approximately 150:1, approximately 140:1, approximately 130:1, approximately 120:1, approximately 110:1, approximately 100:1, approximately 90:1, approximately 80:1 or approximately 70:

1.

6. The cell culture medium of any of claims 1-5, wherein a first stimulant is IL-7, present in a concentration of approximately 100 Ul / ml to approximately 5,000 Ul / ml; and a second stimulant is IL-15, present in a concentration of approximately 1 Ul / ml to approximately 100 Ul / ml.

7. The cell culture medium according to claim 6, wherein IL-7 is present at a concentration of approximately 300 Ul / ml to approximately 5,000 Ul / ml and IL-15 is present at a concentration that is approximately 25 Ul / ml to approximately 50 Ul / ml.

8. The cell culture medium according to claim 7, wherein IL-7 is present at a concentration of approximately 5,000 IU / ml and IL-15 is present at a concentration of approximately 50 IU / ml.

9. The cell culture medium of any of claims 1-8, wherein extracellular potassium is present at a concentration of approximately 20 mM to approximately 35 mM or approximately 20 mM to approximately 30 mM.

10. The cell culture medium according to claim 9, wherein extracellular potassium is present at a concentration of approximately 20 mM or approximately 25 mM.

11. The cell culture medium of any of claims 1-10, wherein extracellular potassium is present as KCI.

12. A method for obtaining a T cell population in vitro, comprising culturing an initial T cell population with a cell culture medium comprising a first and a second cell proliferation stimulant, each independently selected from the group consisting of IL-4, IL-7, IL-10, IL-12, and IL-15; and an extracellular modulator of cell metabolism being extracellular potassium at a concentration of approximately 4 mM to approximately 40 mM, wherein the first and second stimulants are present in a concentration ratio of approximately 1,000:1 to approximately 4:

1.

13. The method of claim 12, wherein the cell culture medium is the cell culture medium of any of claims 1-11.

14. The method of claim 12 or 13, wherein the resulting T cell population is enriched in Tcm and / or Tscm cells.

15. The method of any of claims 12-14, wherein the resulting T cell population comprises at least approximately 30%, 35%, 40% or 45% Tscm cells.

16. The method of claim 15, wherein the resulting T cell population comprises at least approximately 40% Tscm cells.

17. The method of any of claims 12-16, wherein the resulting T cell population is enriched approximately 100 to approximately 1,000 times more in Tcm and / or Tscm than in Tcm and / or Tscm of the initial T cell population measured over a period of approximately 7-16 days.

18. The method of claim 17, wherein the resulting T cell population is enriched approximately 100 to approximately 1,000 times more in Tcm and / or Tscm than in Tcm and / or Tscm of the initial T cell population measured over a period of approximately 10-14 days.

19. The method of claim 18, wherein the resulting T cell population is enriched at least approximately 100 or approximately 200 times more in Tcm and / or Tscm than in Tcm and / or Tscm of the initial T cell population.

20. The method of any of claims 12-19, wherein the initial T cell population is a manipulated T cell population.

21. The method of claim 20, wherein the initial T cell population is a T cell population expressing one or more chimeric antigen receptors.

22. The method of any of claims 12-21, wherein the T cells are allogeneic T cells.

23. The method of any of claims 12-21, wherein the T cells are autologous T cells.

24. A population of manipulated immune cells, wherein said population comprises T cells expressing one or more chimeric antigen receptors (CAR-T cells), and wherein said CAR-T cells are obtained by using the cell culture medium of any of claims 1-11.

25. A population of manipulated immune cells, wherein said population comprises T cells expressing one or more chimeric antigen receptors (CAR-T cells), and wherein said (CAR-T cells) are obtained by using the method of any of claims 12-23.

26. The manipulated immune cell population according to claim 24 or 25, wherein the CAR-T cells comprise at least approximately 30%, 35%, 40%, or 45% Tscm cells- 27. The manipulated immune cell population according to claim 26, wherein the CAR-T cells comprise at least approximately 40% Tscm cells.

28. A pharmaceutical composition comprising the manipulated immune cell population of any of claims 24-27.

29. A method for treating a disease or disorder in a subject in need, comprising administering to the subject the manipulated immune cell of any of claims 24-27, or the pharmaceutical composition according to claim 28.

30. The method of claim 29, wherein the disease or disorder is cancer.

31. A manufactured article comprising the manipulated immune cell of any of claims 24-27 or the pharmaceutical composition according to claim 28.