Figure 2.

(A and B) Day-6 DCs generated with or without D₃ analog from VDR WT and VDR KO mice were plated in equal numbers for 48 h in the presence or absence of anti-CD40. Flow cytometric analysis was carried out 48 h later and expressed as median fluorescence intensity (MFI) for CD80/CD86 of CD11c^+ve cells (A). ELISA for IL-12 p40 was carried out on culture supernatants 48 h later (B). Both WT and KO unconditioned DCs demonstrated increased CD80/CD86 expression and IL-12 p40 secretion with anti-CD40. D₃ analog-conditioned WT DCs failed to significantly up-regulate CD80/CD86 expression and IL-12 p40 secretion after CD40 cross-linkage whereas D₃ analog-conditioned KO DCs responded similarly to unconditioned DCs. (C and D) Day-6 unconditioned or D₃ analog-conditioned DCs from B6 mice were cultured for 48 h without additional stimulation and with two concentrations each of MCM (C, 12.5% and 50% final volume) or LPS (D, 10 and 50 ng/ml). Results are shown of MFI for CD80/CD86 by CD11c^+ve cells. Unconditioned DCs responded to the lower concentration of both stimuli by significantly increasing CD80/CD86 expression. D₃ analog-conditioned DCs had lower basal expression of CD80/CD86 that did not significantly increase even with the higher concentrations of the two stimuli. *, P < 0.05 compared with unstimulated cells from the same population; †, P < 0.05 compared with unconditioned DCs.