Fig. 2

Activation, proliferation and cytotoxicity of Tan-CAR T cells. CAR T-cell activation was evaluated by flow cytometric identification of cell surface expression of CD69. CD69 detected on tan-CAR T cells after co-culture with K562 cells overexpressing CD19, BCMA, or both at levels that were comparable to those of the single scFv-CAR T cells co-cultured with target cells expressing their cognate antigens (n = 3). B) Proliferation in response to mitomycin treated K562-transfectant target tumor cells. CAR T cells were labeled with CFSE prior to 5-day co-culture with replication-incompetent target cells. T-cell proliferation was evaluated by flow cytometry. (C)&(D) CAR T-cell-mediated cytotoxicity was evaluated by quantitative assessment of LDH in the supernatants of co-cultured cells as indicated; data are presented as the mean ± SD, *p < 0.05; **p < 0.01; ***p < 0.001 vs. un-transduced T cells (NC) from the same donor