Migration and cytokine secretion by mature MoDCs differ in response to differentially polymerized CD40L preparations. MoDCs were generated from CD14⁺ monocytes within 5 to 7 days of culture in the presence of GM-CSF and IL-4. Immature MoDCs (1 × 105 to 3 × 105/mL) were washed and resuspended in culture medium. The following cytokines were added as indicated for an additional 36 to 48 hours: GM-CSF (20 ng/mL), TNF-α (10 ng/mL), IFN-α (2000 IU/mL), PGE₂ (1 μM), CD40L₁ ([CD40L monomers] 1 μg/mL; enhancer, 1 μg/mL), CD40L₃ ([CD40L trimers] 1 μg/mL), and IFN-γ (1000 U/mL). (A) Migration assays were performed in duplicate using 3 μm pore size transwell plates. Migration to medium in the absence or presence of CCL21 (6Ckine, 40 ng/mL) in the lower well is shown as mean values ± SEM of 14 experiments. Supernatants were harvested and (B) IL-12p70 and (C) IL-6 production measured by ELISA (mean values ± SEM, n = 11). *P < .05; **P < .01 as compared with activation with CD40L₃ plus IFN-γ.