(A) The final product of CAR-expressing T cells showed mainly central memory and effector memory T cells regardless of TCR expression after expansion (n=3). (B) Surface expression profile of several co-stimulatory (OX40, CD28, 4-1BB) and co-inhibitory molecules (PD-1, TIM-3, BTLA, CTLA-4, TIGIT, VISTA) as well as commonly used exhaustion markers (2B4, LAG-3, PD-1) was determined by flow cytometry and compared between CAR-T cells with or without TCR KO (n=3). (C) Cellular composition of CAR-T cells was determined on day 14 (n=4). A two-tailed paired t test was performed to determine statistical significance. Tn: naïve T cells, Tscm: stem cell-like memory T cells, Tcm: central memory T cells, Tem: effector memory T cells, Teff: Effector T cells; Tn: CD62L+, CD45RO-, CD95-; Tscm: CD62L+, CD45RO-, CD95+; Tcm: CD62L+, CD45RO+, CD95+; Tem: CD62L-, CD45RO+, CD95+; Teff: CD62L-, CD45RO-, CD95+, RV19BB_TCR+=conventional CAR-T cells without electroporation, RV19BBCRTCR+=CAR-T cells electroporated with nonsense gRNA, RV19BBCRTRBC-=CAR-T cells electroporated with TRBC-targeting gRNA, n.s.=not significant, NKT cells=natural killer T cells.