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. 2021 May 27;14(8):101138. doi: 10.1016/j.tranon.2021.101138

Fig. 2.

Fig. 2

Antigen-dependent cytotoxicity of CAR-T cells. (A) FACS analysis of CD276 expression on ESCC cell lines. (B) Structures of CAR used. (C-E) T cells from healthy donors were transduced with CAR via lentivirus and expanded. Then numbers of CAR-T cells were counted at indicated intervals (C). On day 12, CAR transduction efficacy (D) and memory differentiation (E) were determined by FACS. t-test was performed. (F) BLI detection of the viability of tumor cells incubated with indicated CAR-T cells at the listed E/T ratios without exogenous cytokines. (G) Effector cytokines secretion of CAR-T cells at E/T = 1 without exogenous cytokines. (H and I) The FFluc-expressing TE-7 cell line was engineered to express CD276 via lentiviral transduction. Cells were sorted based on CD276 levels and single clones were expanded (H). Then FFluc+ TE-7 (wildtype) and the derivate CD276+ clones were co-cultured with CAR-T cells at E/T = 1 without exogeneous cytokines. Tumor cell viability was detected by BLI after 24 h (I). Data represent assays using CAR-T cells produced from five donors. *, P < 0.05.