Analysis of migration and cytokine secretion by MoDCs or autologous FL-generated PBDCs.
PBDCs were purified by negative depletion from the peripheral blood of melanoma patients treated with FL for 14 consecutive days, and then FACS were sorted to high purity (> 95%) on the basis of CD1b/c and HLA-DR expression. Autologous MoDCs were generated from blood isolated from pretreatment bleeds and cultured 7 days prior to the isolation of PBDCs. (A) Migration toward CCL19 by immature MoDCs or MoDCs stimulated with the indicated stimuli for 48 hours. (B) Migration toward CCL19 by sorted FL-generated PBDCs from melanoma patients with minimal residual disease stimulated with the indicated stimuli for 48 hours. (C) IL-12p70 secretion by immature MoDCs or MoDCs stimulated (1 × 105/well) with the indicated stimuli for 3 days. (D) IL-12p70 secretion by FL-generated PBDCs (1 × 105/well) from melanoma patients with minimal residual disease stimulated with the indicated stimuli for 3 days. Culture SNs were examined for cytokine production by ELISA. Data represent the means ± SEM of experiments from 7 separate donors.
