Figure 5.

Nutlin-3 or 10058-F4 do not significantly affect the cytotoxic function of activated T cells. (A) 10⁵ T cells pre-treated with nutlin-3, 5 µM (gray bars) or DMSO-solvent control (black bars) for 48 h were washed and then activated with PHA (1 µg/ml) and pulsed with ³H-thymidine for 16 h at the indicated time points. (B) Cytotoxic assay – Killing of chromium-51 labeled allogeneic LCL by T cells activated for 24 h following a 48 h pre-treatment with nutlin-3, 5 µM (broken line) or DMSO-solvent control (solid line). (C) p53 expression in activated T cells pre-treated with nutlin-3 (5 µM) or DMSO for 48 h, washed and activated by by PHA (1 µg/ml) for 24 h. (D) 10⁵ T cells were activated for 48 h and cultured with nutlin-3 (5 µM) or 10058-F4 (50 µM) simultaneously (black bars) or alternatively cells were cultured with each compound only for the last 24 h (gray bars). Cells treated with DMSO served as control. (E) Cytotoxic assay - Killing of chromium-51 labeled allogeneic LCL by activated T cells treated with nutlin-3 5 µM (broken line) or 10058-F4, 50 µM (dotted line) in parallel with PHA and (F) for the last 24 h of the 48 h activation. Cells treated with DMSO served as control.