Figure 5.

Generation and Characterization of hPSMA-Directed CAR T Cells

(A) hPSMA-specific CAR receptor and hPSMA-specific CAR-encoding retroviral vector design. (B) FACS profiling of hPSMA-directed CAR T cells: (1) hPSMA CAR-transduced cells (right upper quadrant of (a) was selected for further analysis; (2) a higher percentage of CD8⁺ T cells over CD4⁺ T cells was observed (b); (3) the predominant population was an effector-memory RA subset (T_EMRA) (CD45RA⁺/CCR7⁻) in both CD8⁺ (dominant) and CD4⁺ T cells (c and d, respectively) after stimulation with NIH 3T3 hPSMA⁺ cells. T_N, naive T cells; T_EM, effector memory T cells; T_CM, central memory T cells. (C) Expansion of CAR T cells in the presence of increasing concentrations of Sodium lactate. (D) Viability of CAR T cells in the presence of increasing concentrations of Sodium lactate. (E) Comparison of in vitro⁵¹Cr-chromium-release assays. A greater toxic effect of CAR T cells was observed on the LDH-A KD cell line as compared to the NC and WT cell lines (p = 0.0005 and p < 0.0001, respectively).