Up-regulation of CCR7 and MIP-3β expression in IFN-DCs and CD83⁺ cell migration in response to exogenous MIP-3β.

(A) RT-PCR analysis showing the up-regulation of CCR7 and MIP-3β mRNAs in IFN-DCs as compared with IL-4-DCs. (B) Strong migratory capacity of IFN-DCs in response to MIP3β. Migration assays were performed in a 24-well Transwell cell culture chamber (Costar). MIP3α and MIP3β chemokines (100 ng/mL) were loaded in the lower compartment. After 2 hours of incubation, the cells that migrated in the lower compartment were collected and counted. Assays were performed in triplicate. SDs did not exceed 15%. (C) The majority of IFN-DCs migrated in response to MIP-3β expressed the CD83 marker. After migration assay, cells recovered from the lower compartment of the Transwell chambers were collected and analyzed by flow cytometry. MIP-3β-responsive cells in IFN-DCs were mostly CD83⁺, whereas the few IL-4-DCs migrated in response to MIP-3β did not express CD83. The addition of MIP3-β to the cell cultures did not induce up-regulation of CD83 (data not shown). The results are representative of 3 experiments performed with cells from different donors.