Fig. 4.

Reproducibility of triple CAR T-cell transduction achieved with TVP derived virus. (A) Representative histograms showing the median fluorescent intensity of CAR marker expression in anti-CD38 + CLEC12A + TIM3 (38/C/T) CAR T-cells and control untransduced T-cells in gray. (B) Percentages of triple CAR positive (TP), double CAR positive (DP), single CAR positive (SP) or triple negative (TN) T-cells from three distinct donors transduced with three independent virus batches (B1-3). (C, D) Variability in 38/C/T CAR T-cell population composition after transduction between healthy primary T-cell donors (D1-3) or virus batches (B1-3). In all experiments n = 3 and duplicates, error bars represent mean ± SD. In (B) different symbols each represent a different T-cells donor. In this figure, CAR1 (dsRed) is a previously described high affinity (A1) second generation anti-CD38 CAR with a CD28 costimulatory domain,²⁹ transmembrane domain and hinge, CAR2 (LNGFR) is an anti-TIM3 CAR and CAR3 (sII) is an anti-CLEC12a CAR, both containing a CD8a hinge and transmembrane domain and 4-1BB costimulatory domain.