Figure 5

Selective activation of Vγ2Vδ2 T cells by compound 7. (A) Selective expansion of Vγ2Vδ2 T cells from blood αβ and γδ T cells after culture with compound 7. PBMC from a prostate cancer patient were stimulated with 1 μM compound 7 and IL-2. The two-color flow cytometric analysis of Vγ2Vδ2 T cells in PBMC before (left panel) and after (right panel) 10 day stimulation is shown. (B) Inhibition of EJ-1 bladder carcinoma cell growth by exposure to compound 7 and γδ T cells. Compound 7 was added to cultures of EJ-1 tumor cells followed by the addition of Vγ2Vδ2 T cell to some cultures16 h later. Cell growth was assessed in a real-time cell analyzer system. Culture conditions were: (1) 50 nM compound 7 + medium, (2) 0 nM compound 7 + γδ T cells, (3) 1.56 nM compound 7 + γδ T cells, (4) 12.5 nM compound 7 + γδ T cells, (5) 25 nM compound 7 + γδ T cells, (6) 50 nM compound 7 + γδ T cells. (C) Degranulation of γδ T cells in response to U937 histocytoma pretreated with compound 7. The proportion of CD107a⁺ degranulated Vδ2⁺ cells were plotted against the concentrations of compound 7 used for the pretreatment of U937 cells. (D) Stimulation of IFN-γ production by Vγ2Vδ2 T cells by compound 7. PBMC from healthy donor were cultured with 1 μM compound 7. After 48 h, the culture supernatants were removed and IFN-γ levels determined by ELISA.