Independent regulation of migration and cytokine secretion by cAMP (cBIMPS), PI3K, and PC-PLC/PLD: E coli –induced DC activation. MoDCs were activated using intact E coli in the presence or absence of inhibitors. Inhibitors were added to MoDCs 20 to 40 minutes prior to E coli exposure. Wortmannin and cAMP were used as in Figure 1. D609 (tricyclodecan-9-yl xanthate potassium) was used at a concentration of 300 μM. Cells and supernatants were harvested 36 to 48 hours later. (A) Migration toward CCL21 (means ± SEM, n = 8-9 separate donors; *P < .05, **P < .01 compared with activation without inhibitor). Secretion of IL-6 (B) and IL-12p70 (C) measured by ELISA (mean values ± SEM, n = 5-12 separate donors for IL-6 and n = 10-13 for IL-12p70; *P < .05, **P < .01 compared with activation without inhibitor). (D) MFI of CD80, CD83, and CD86 expression for MoDCs activated with E coli in the presence of inhibitors relative to MoDCs activated without inhibitors (means ± SD, n = 4-6 separate donors; *P < .05, **P < .01 compared with activation without inhibitors). (E) Schematic of the independent regulation of migration and IL-12p70 secretion of DCs. Both functions are enhanced by p38K and PC-PLC, whereas the PI3K/Akt-1 pathway inhibited only IL-12p70 secretion. In contrast, cAMP enhanced migration and inhibited IL-12p70, whereas ERK1/2 inhibited migration and enhanced IL-12p70.