Fig. 3

The strategy for enhancing Vγ9Vδ2 T cell therapy in the treatment of glioma in the WAT group. Vγ9Vδ2 T cells were incubated with WAT primary glioma cells (TT-LS-luc, TT-YZJ-luc, TT-ZLH-luc, TT-XJ-luc, TT-SX-luc, and TT-HCL-luc) at E:T ratios of 1:1 in the presence or absence of ZOL (5 μM) or BTN3A1 agonistic antibody (BTN3A1 20.1, 1 µg/mL) for 18–20 h. The killing of Vγ9Vδ2 T cells against WAT primary glioma cells was significantly enhanced after ZOL stimulation A or BTN3A1 agonistic antibody stimulation B. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Car-B7H3-γδT cells had stronger and wider anti-glioma ability than parental Vγ9Vδ2 T cells in vitro. (C) CAR construct is shown. (D)Vγ9Vδ2 T cells and Car-B7H3-γδT cells were co-cultured with U87-B7H3-KO cells in a 1:1 ratio for 18–20 h in the presence or absence of ZOL (5 μM). Luciferase activity was measured. Data are expressed as the mean ± SD. E Flow cytometry analysis was performed to determine B7-H3 expression in WAT primary glioma cells. The expression of B7-H3 protein was higher than 90%. F Vγ9Vδ2 T cells and Car-B7H3-γδT cells were incubated with WAT primary glioma cells at E:T ratios of 1:1 for 18–20 h. Luciferase activity was measured. Data are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. G, H The culture medium was collected, and ELISA was performed to measure IFN-γ and TNF-α levels. Data are expressed as the mean ± SD. *P < 0.05, ***P < 0.001, ****P < 0.0001; n = 3