Fig. 2

Pep42-BBZ CAR-T recognized GRP78 and kill glioma cell lines. A Schematic showed the structure of mock-BBZ and Pep42-BBZ CARs. B Quantification of the percentage of CAR-positive T cells 96 h post transduction detected by flow cytometry. Left: Statistics of transduction efficiency of Pep42-BBZ CAR-T group and mock-BBZ CAR-T group from three healthy donors. Right: Representative images from one donor of flow cytometry. The results were from three independent experiments. NTD = non-transduced T cell controls. C The proliferation fold change of the three groups of T cells from day 0 to day 17 post lentiviral construct transduction. D Measured VCN per cell of Pep42 CAR-T cells derived from two of the three healthy donors above. Cells were assessed 96 h post transduction and the transduction MOI and efficiency were the same as the above. E Cytotoxicity ratios of mock-BBZ and Pep42-BBZ CAR-T cells to U-251MG and U-87MG cells at increasing E:T ratios of 1:1, 2:1, 4:1 and 8:1. CAR-T cells were co-cultured with target cells for 20 h. Results were calculated based on the lysis ratio of NTD T cells to the target cells at every E:T ratio. F IFN-γ release levels from the supernatant of cells co-cultured at an E:T ratio of 8:1 quantified by ELISA. All effector cells used in the cytotoxicity assay were based on total T cells in each group. Data presented as the mean volume ± SD, * P < 0.05, ** P < 0.01 *** P < 0.001