Up-regulation of maturation markers on MoDCs in response to proinflammatory factors.

MoDCs were prepared by culturing purified CD14⁺ monocytes for 7 days in GM-CSF and IL-4 and stimulated for 3 days with the indicated stimuli. Surface expression of maturation markers was examined by flow cytometry on day 10. (A) Immature MoDCs were stimulated with either TNF-α (20 ng/mL) or IFN-α (1000 IU/mL) or PGE₂ (1 μM) or combinations thereof as indicated for 3 days, and flow cytometric analysis of CD83 expression was performed. (B) Immature MoDCs were stimulated for 3 days with either TNF-α + IFN-α + PGE2 or CD40L (1 μg/mL) or intact E coli(1 × 10⁶), and flow cytometric analysis of CD80, CD83, CD86, HLA-I, and HLA-II expression was assessed. Results are shown as fold increase of mean fluorescence levels relative to nonstimulated control MoDCs (control, normalized to 1). Figures represent the means (SEM of 4 experiments for (C) FITC-dextran uptake or (D) PE-latex bead uptake (1 μm) at either 4°C or 37°C for 30 minutes. Cells were examined by flow-cytometry to assess internalized FITC or PE. The data are presented as the MFI of internalized FITC or PE and represent means (SEM) of 4 experiments.