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. 2020 Sep 13;12(9):2612. doi: 10.3390/cancers12092612

Figure 4.

Figure 4

In vitro cytotoxicity of iKP-19-CAR-T cells against CD19+HLA-C1− Daudi cells or CD19+HLA-C1+ normal B cells. (A) Representative CD19 and HLA-C1 expression in B cells. (B–E) After CD19 CAR rate was unified, iKP-19-CAR-T cells and CD19-CAR-T cells were co-cultured with Daudi cells or normal B cells at a 1:1 ratio for 6 h in RPMI-1640 medium. The expressions of activation marker CD69 (B), degranulation marker CD107a (C), GzmB (D), and signal molecule P-Zap70 (E) in CAR-T cells was detected by flow cytometry using PE-anti-human CD69 antibody, PE-anti-human CD107a antibody, PE-anti-human GzmB antibody and PE-anti-human P-Zap-70 antibody. MFI was statistically measured from three independent experiments (n = 4 different donors). (F) LDH assay was performed to test the cytotoxicity of iKP-19-CAR-T cells and CD19-CAR-T cells against Daudi cells or normal B cells after 6h co-culture at different E:T ratio (n = 4 different donors). All experiments were performed in triplicate manner using PBMCs of every donor. ** p < 0.01, *** p < 0.001. Error bars represent ± SD.