In particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway inhibits PDK-1 activation of AKT. In various embodiments, a method for maintaining the proliferation and decreasing the differentiation of restimulated T cells expressing an engineered TCR or CAR comprising: (a) contacting all or a portion of a population of proliferated T cells comprising an engineered TCR or CAR with an anti-CD3 antibody or CD3-binding fragment thereof, and an anti-CD28 antibody or CD28-binding fragment thereof, which stimulates a CD28 accessory molecule on the surface of the T cells, thereby restimulating the activated T cells to proliferate; wherein the restimulated T cells have maintained proliferation and decreased differentiation compared to the proliferation of T cells that were stimulated or restimulated in the absence of the inhibitor of the PI3K/AKT/mTOR pathway is provided. In additional embodiments, the population of activated T cells restimulated in the presence of an inhibitor of PI3K/AKT/mTOR pathway do not express CD57 or KLRG1 or express less CD57 or KLRG1 compared to a population of T cells activated and stimulated in the absence of the inhibitor of PI3K/AKT/mTOR pathway.



In particular embodiments, the population of activated T cells restimulated in the presence of an inhibitor of PI3K/AKT/mTOR pathway have an increased number of T cells expressing one or more markers selected from the group consisting of: CD62L, CCR7, CD28, CD27, CD122, and CD127 compared to a population of T cells activated and stimulated in the absence of the inhibitor of PI3K/AKT/mTOR pathway. In additional embodiments, the inhibitor of the PI3K/AKT/mTOR pathway inhibits HSP activation of AKT. In further embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a pan-mTOR inhibitor selected from the group consisting of: AZD8055, INK128, and rapamycin. In other particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a pan-mTOR inhibitor selected from the group consisting of: AZD8055, INK128, and rapamycin. In particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway inhibits HSP activation of AKT. In other particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a DNA-PK inhibitor selected from the group consisting of: NU7441, PI-103, NU7026, PIK-75, and PP-121.



In particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is selected from the group consisting of: BEZ235, LY294002, GDC-0941, BYL719, GSK2636771, TGX-221, AS25242, CAL-101, IPI-145, MK-2206, GSK690693, GDC-0068, A-674563, CCT128930, AZD8055, INK128, rapamycin, PF-04691502, everolimus, BI-D1870, H89, PF-4708671, FMK, AT7867, NU7441, PI-103, NU7026, PIK-75, ZSTK474, and PP-121. In some embodiments, the inhibitor of the PI3K/AKT/mTOR pathway inhibits DNA-PK activation of AKT. In additional embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is the selective AKT1 inhibitor A-674563. In particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a s6 kinase inhibitor selected from the group consisting of: BI-D1870, H89, PF-4708671, FMK, and AT7867. In some embodiments, watch live sexcam the inhibitor Https://Watchlivesexcam.Com of the PI3K/AKT/mTOR pathway is a s6 kinase inhibitor selected from the group consisting of: BI-D1870, H89, PF-4708671, FMK, and AT7867. In some embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is selected from the group consisting of: BEZ235, LY294002, GDC-0941, BYL719, GSK2636771, TGX-221, AS25242, CAL-101, IPI-145, MK-2206, GSK690693, GDC-0068, A-674563, CCT128930, AZD8055, INK128, rapamycin, PF-04691502, everolimus, BI-D1870, H89, PF-4708671, FMK, AT7867, NU7441, PI-103, NU7026, PIK-75, ZSTK474, and PP-121. In various embodiments, a population of T cells comprising a vector comprising an engineered TCR or CAR, wherein the cells have been activated and stimulated to proliferate in the presence of an inhibitor of PI3K/AKT/mTOR pathway is provided.



In further embodiments, the population of T cells activated and stimulated in the presence of an inhibitor of PI3K/AKT/mTOR pathway do not express CD57 or KLRG1 or express less CD57 or KLRG1 compared to a population of T cells activated and stimulated in the absence of the inhibitor of PI3K/AKT/mTOR https://Watchlivesexcam.com/ pathway. In particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a DNA-PK inhibitor selected from the group consisting of: NU7441, PI-103, NU7026, PIK-75, and PP-121. In certain embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a pan-AKT inhibitor selected from the group consisting of: MK-2206, GSK690693, and GDC-0068. In particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a pan-AKT inhibitor selected from the group consisting of: MK-2206, GSK690693, and GDC-0068. In certain embodiments, the inhibitor of the PI3K/AKT/mTOR pathway inhibits mTORC2 activation of AKT. In certain embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a selective mTORC1 inhibitor selected from the group consisting of: PF-04691502 and everolimus.



In other particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a selective PI3K inhibitor selected from the group consisting of: BYL719, GSK2636771, TGX-221, AS25242, CAL-101, and IPI-145. In additional embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is the selective AKT2 inhibitor CCT128930. In other particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is the PI3K inhibitor ZSTK474. In other particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway the PI3K inhibitor ZSTK474. In certain embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is the selective AKT2 inhibitor CCT128930. In further embodiments, the inhibitor of the PI3K/AKT/mTOR pathway inhibits PDK-1 activation of AKT. In particular embodiments, wherein the inhibitor of the PI3K/AKT/mTOR pathway is the selective AKT1 inhibitor A-674563. In some embodiments, the population of T cells activated and stimulated in the presence of an inhibitor of PI3K/AKT/mTOR pathway have an increased number of T cells expressing one or more markers selected from the group consisting of: CD62L, CCR7, CD28, CD27, CD122, and CD127 compared to a population of T cells activated and stimulated in the absence of the inhibitor of PI3K/AKT/mTOR pathway. In certain embodiments, the inhibitor of the PI3K/AKT/mTOR pathway inhibits DNA-PK activation of AKT. In particular embodiments, the inhibitor of the PI3K/AKT/mTOR pathway is a pan-PI3K inhibitor selected from the group consisting of: BEZ235, LY294002, and GDC-0941.