Fig. 4.

a–c Functional characterization of tumor cell-treated DC. a Endocytic activity of DC differentiated in the presence of CM from tumor cell lines. Ctrl-DC, A-427-DC, Calu-1-DC, and MΦ were incubated with FITC-dextran at 37°C or at 4°C as a control of internalization, and then analyzed by flow cytometry. The experiment shown is representative of four similar performed ones. Values to the right are the mean ± SD of FITC incorporation of the independent experiments. b Cytokine mRNA detection on the above cell populations. RT-PCR for the following molecules was performed: TNF-α (lane 1), IL-10 (lane 2), IL-6 (lane 3), TGF-β1 (lane 4), and β-actin (lane 5). Shown is an agarose gel electrophoresis of RT-PCR fragments in a representative experiment out of five. c Cytokine secretion by Ctrl-DC (white bars), A-427-DC (black bars), Calu-1-DC (gray bars), and MΦ (hatched bars). Cells were stimulated with LPS plus IFN-γ, and the supernatants were harvested after 24 h. Production of IL-6, IL-10, and IL-12 was evaluated by ELISA. Data shown are the mean ± SD of four independent experiments. Statistical analysis (MΦ or tumor cell-treated DC versus Ctrl-DC): ^* p<0.05