Figure 6
ReoTCM effectively supports priming of specific CTL by tumour cell-loaded DC. PBMC were incubated with autologous DC that had been cultured overnight with Mel-888 tumour cells in the presence of reoTCM/non-reoTCM. The PBMC were restimulated 7 days later and assayed at 14 days. (A) Lymphocyte proliferation was determined via trypan blue exclusion (2 representative donors are shown). (B) IFN-γ levels in CTL supernatants were determined by ELISA (2 representative donors are shown). (C) Cytotoxicity of lymphocytes primed in the presence of reoTCM versus non-reoTCM was determined by 51Cr release assay using Mel-888 tumour cells as specific targets and SKOV-3 as irrelevant controls. One donor is shown as representative of 2 independent experiments. * indicates P < 0.05 by Student's t-test. (D) CTL as in (C) were further assayed for CD107 degranulation and intracellular IFN-γ on co-culture with Mel-888 targets. The results of one donor are shown, representative of at least four independent experiments.