Fig. 2

Generation of B7-H3 CAR- T cells and their in vitro antitumor activity. a Schematic diagram of the CAR construct. CAR gene was cloned into a lentiviral vector, which contained an internal ribosome entry site (IRES)-green fluorescence protein (ZsGreen) sequence. The resulting CAR was composed of the leader sequence (LS), a scFv, a hinge (H) region, a CD8α transmembrane domain (TM), along with the 4-1BB domain and the CD3ζ chain. b Subsets of B7-H3 CAR-T cells were derived from a single healthy donor. T cells were stained with anti-CD3, anti-CD4, or anti-CD8 antibodies and then analyzed using flow cytometry. c Proliferation of B7-H3 CAR-T cells and vehicle T cells activated by anti-CD3/CD28 beads. d Cytotoxicity of B7-H3 CAR-T cells (red line) and vehicle T cells (blue line) cells against various tumor cell lines. e Apoptosis in target A549 cells induced by B7-H3 CAR-T cells and vehicle T cells as control at different E:T ratios. Representative graphs are shown. The p values of the difference between the CAR group and the control group were analyzed using ANOVA