scFV-3H11-CAR-T cells mediate robust cytotoxicity, and IL-2 and IFN-γ production against antigen positive gastric tumor cells. (A) Flow cytometry histogram plots of 3H11-CAR lentivirus transduction efficiency for 3H11-CAR in Jurkat cells using anti-Myc antibody staining. Cytotoxicity assays were performed to measure the specific lysis of (B) BCG-823, (C) NCI-N87 and (D) GES1 cells by scFV-3H11-CAR T Jurkat cells or mock Jurkat cells for 8 h at different E/T ratio. (E) Flow cytometry histogram plots of 3H11-CAR lentivirus transduction efficiency for 3H11-CAR in CD3+ T cells from healthy donors using anti-Myc antibody staining. Cytotoxicity assays were performed to measure the specific lysis of (F) BCG-823, (G) NCI-N87 and (H) GES1 cells by scFV-3H11-CAR T cells or mock T cells for 8 h at different E/T ratios. Measurements of IFN-γ production in (I) BCG823 and (J) NCI-N87 cells at different E/T ratios for 8 h, as determined by ELISA. Measurements of IL-2 production in (K) BCG823 and (K) NCI-N87 cells at different E/T ratios for 8 h, as determined by ELISA. Data in (B-D and F-L) are presented as the mean ± standard error of the mean from three independent experiments with triplicate wells. Data in (B-D and F-L) were analyzed by one-way analysis of variance. CAR, chimeric antigen receptor; IL-2, interleukin 2; IFN-γ, interferon γ; ns, not significant.