FIG 5.
Forced maturation induces CCR7 expression and CCL19-directed migration of P. gingivalis-infected MoDCs. MoDCs that had been infected as indicated for 24 h were treated with E. coli LPS/TNF-α for an additional 24 h to force maturation. (A) Representative histograms are shown for each MoDC infection treatment for CCR7 expression. The inflammatory cocktail leads to significant upregulation of surface CCR7 expression. (B) Summary data for CCR7 expression are shown, representing the additional 24-h treatment with the inflammatory cocktail. The treatment causes significant upregulation of CCR7 in all of our infected MoDCs, which matches with what occurs in typical matured MoDCs. (C) After treatment with inflammatory cocktail, cells were collected and loaded into the upper compartment of a microchemotaxis Boyden chamber (1 × 105/well) and migrated toward a CCL19 chemokine gradient. Migration distance (μm) was determined with leading-edge analysis after hematoxylin staining by using a 40× objective lens and subtracting migration distance without chemokine from migration distance with chemokine. The infected MoDCs show an increased response to CCL19 after the additional treatment compared to untreated controls. *, P < 0.01.