Extended Data Fig. 4: BATF overexpression improves effector function in human CAR T cells.

a, Histograms showing expression of human BATF (hBATF, left panel) and human CAR (hCAR [stained by Goat Anti-Armenian Hamster IgG (H + L)], right panel) in the corresponding lentivirally transduced human CD8⁺ T cells. FMO, fluorescence-minus-one control; TIG, control cells transduced with the empty vector. b, Left, Histograms showing CellTrace Violet (CTV) dilution in lentivirally transduced human CD8⁺ CAR T cells. Right, Proliferation index calculated as (Day 0 CTV MFI) / (Day 4 CTV MFI). c, Histograms showing expression of the indicated cytokines after resting in X-Vivo media or after stimulation with PMA and ionomycin for 4 h. d-e, Human CAR T cells were labeled with CellTrace Violet and co-cultured with NALM6 cells for 5 h. d, Gating strategy for in vitro cytotoxicity assay of control (TIG) or hBATF-transduced human CAR T cells. e, Histograms showing ratio between target cells (NALM6) and effector cells (human CAR T cells). Percent cytotoxicity was calculated as (1- (R5/R0)) x 100, where R5 = (target cells as % of total at 5 h) / (effector cells as % of total at 5 h), R0 = (target cells as % of total at 0 h) / (effector cells as % of total at 0 h). Each dot in b-e represents an individual donor. Data were obtained from four biological experiments and analyzed by one-tailed unpaired Student’s t-test.