Independent regulation of migration and cytokine secretion by cAMP (cBIMPS) and PI3K: CD40L-induced DC activation. Immature MoDCs were washed and resuspended in culture medium at a concentration of 1 to 3 × 105 cells/mL. BHK-CD40L cells were irradiated and added at a concentration of 1:20 to MoDCs either alone or together with PGE₂ (1 μM), wortmannin (100 ng/mL), and the cAMP analog and activator of protein kinase A (Sp-5,6-DCl-cBIMPS; 50 μM). Supernatants and cells were harvested after 36 to 48 hours. (A) Secretion of IL-6 and IL-12p70 measured by ELISA (means ± SEM of 7 experiments; *P < .05, **P < .01 compared with activation with the BHK-CD40L cell line). (B) Mean fluorescence intensity (MFI) of CD80, CD83, and CD86 expression for MoDCs activated with the BHK-CD40L cell line in the presence of inhibitors relative to MoDCs activated without inhibitors (means ± SD, n = 6-16 separate donors; *P < .05, **P < .01 compared with activation with the BHK-CD40L cell line).(C) Migration toward CCL21 (means ± SEM, n = 5-8 separate donors; *P < .05 compared with activation with the BHK-CD40L cell line).