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In various embodiments, a method for manufacturing T cells comprising: (a) isolating a population of T cells, e.g., tumor infiltrating cytotoxic T lymphocytes (TIL), from a subject; (b) activating the population of T cells and stimulating the population of T cells to proliferate, wherein the activation and stimulation steps are performed in the presence of an inhibitor of AKT/mTOR pathway; (c) culturing the T cells to proliferate; wherein the activating and stimulating steps performed in the presence of the inhibitor of the PI3K/AKT/mTOR pathway results in maintaining proliferation of the T cells compared to the proliferation of T cells that were activated and stimulated in the absence of the inhibitor of the PI3K/AKT/mTOR pathway is provided. In various embodiments, a method for manufacturing T cells comprising: (a) activating a population of T cells and stimulating the population of T cells to proliferate, wherein the activation and stimulation steps are performed in the presence of an inhibitor of AKT/mTOR pathway; (b) transducing the T cells with a viral vector comprising an engineered T cell receptor (TCR) or a chimeric antigen receptor (CAR); (c) culturing the transduced T cells to proliferate; wherein the activating and stimulating steps performed in the presence of the inhibitor of the PI3K/AKT/mTOR pathway results in maintaining proliferation of the transduced T cells compared to the proliferation of transduced T cells that were activated and stimulated in the absence of the inhibitor of the PI3K/AKT/mTOR pathway is provided.



7. The method of claim 1, wherein the viral vector comprises the polynucleotide encoding the CAR. 10. The method of claim 8, wherein the transmembrane domain is derived from CD8.alpha. 14. The method of claim 8, further comprising a signal peptide. 19. The method of claim 16, wherein the PI3K inhibitor is the PI3-K inhibitor ZSTK474. 16. The method of claim 1, wherein the PI3K inhibitor is selected from the group consisting of: BEZ235, LY294002, GDC-0941, BYL719, GSK2636771, TGX-221, AS25242, CAL-101, IPI-145, and ZSTK474. 18. The method of claim 16, wherein the PI3K inhibitor is a selective PI3K inhibitor selected from the group consisting of: BYL719, GSK2636771, Watchlivesexcam.Com TGX-221, AS25242, CAL-101, and IPI-145. NY-ESO-1, HPV, IL-11R.alpha., IL-13R.alpha.2, Lambda, Lewis-Y, Kappa, Mesothelin, Muc1, Muc16, NCAM, NKG2D Ligands, NY-ESO-1, PRAME, PSCA, PSMA, ROR1, SSX, Survivin, TAG72, TEMs, and VEGFR2; a transmembrane domain derived from a polypeptide selected from the group consisting of: CD8.alpha.; CD4, CD28, CD45, PD-1, and CD152; one or more intracellular costimulatory signaling domains selected from the group consisting of: CD28, CD54 (ICAM), CD134 (OX40), CD137 (41BB), CD152 (CTLA4), CD273 (PD-L2), CD274 (PD-L1), and CD278 (ICOS); and a CD3.zeta. In further embodiments, the one or more costimulatory signaling domains are selected from the group consisting of: CD28, CD134, and CD137.



11. The method of claim 8, wherein the one or more costimulatory signaling domains selected from the group consisting of: CD28, CD134, and CD137. 17. The method of claim 16, wherein the PI3K inhibitor is a pan-PI3K inhibitor selected from the group consisting of: BEZ235, LY294002, and GDC-0941. 12. The method of claim 8, further comprising a hinge region polypeptide. 13. The method of claim 12, wherein the hinge region polypeptide comprises a hinge region of IgG1 or CD8.alpha.. 5. The method of claim 1, wherein the viral vector is a retroviral vector. In particular embodiments, the vector is a retroviral vector. 6. The method of claim 1, wherein the viral vector is a lentiviral vector. 3. The method of claim 1, wherein activation of the T cells comprises contacting the T cells with an anti-CD3 antibody or CD3-binding fragment thereof. In certain embodiments, activation of the T cells comprises contacting the T cells with an anti-CD3 antibody or CD3-binding fragment thereof.



However, most, if not all adoptive immunotherapy strategies require T cell activation and expansion steps to generate a clinically effective, therapeutic dose of T cells. Accordingly, there is a persistent, unmet need for improvements in T cell manufacturing and therapeutic T cell compositions that survive, watch Live Sexcam expand, and persist in vivo. Thus, existing T cell manufacturing processes produce an inferior T cell product that is prone to exhaustion and loss of effector immune cell function. Current technologies for generating therapeutic doses of T cells, including engineered T cells, remain limited by cumbersome T cell manufacturing processes. More particularly, the invention relates to methods of T cell manufacturing that result in with improved survival, expansion, and persistence in vivo. More particularly, the invention relates to methods of T cell manufacturing that result in adoptive T cell immunotherapies with improved survival, expansion, and persistence in vivo. In addition, existing T cell activation/expansion methods are normally coupled with substantial T cell differentiation and usually result in short-lived effects, including short-lived survival and a lack of persistence and lack of in vivo expansion of the transferred T cells.