Figure 5
The effect of tick saliva on migration and expression of CCR5 and CCR7 in DCs. Bone marrow cells derived from C57BL/6 mice were collected and cultured for 7 days in the presence of GM-CSF to allow differentiation into DCs. DCs were then pre-incubated with saliva (diluted 1:30, 1:100 v/v) from A. cajennense for 1 hr. DCs were then stimulated for an additional 18 hours with LPS (100 ng/ml). A Boyden microchamber migration assay was then performed with 2x 106 cells/ml seeded into the upper wells and the buffer control (medium) or chemokines (RANTES and MIP-3β) seeded into the lower wells of the microchamber. The incubation period was 1.5 h in a humidified 5% CO2 incubator at 37°C. The filters were then removed and stained and the number of migrated cells was counted (A). Cultured cells were collected and evaluated for the expression of surface molecules CD11c/ CCR5 (B), CD11c/CCR7 (C). In (A), bars represent the mean ± SD number of migrated cells in 15 high power fields from duplicate experiments; in (B) and (C), bars represent the mean ± SD percentage of DCs expressing molecular markers from duplicate experiments. *: p <0.05 compared with DCs cultured with LPS.