Dermal DCs infected with L. major display neutrophil markers.

Mice were treated with control (GL113) or neutrophil-depleting (RB6-8C5 or 1A8) monoclonal antibodies 1 d before challenge in the ear dermis with 2×105 Lm-RFP metacyclic promastigotes. (A) Representative dot plots of CD11b+GR-1hiLy6Cint neutrophils (region 1) and CD11b+GR1intLy6Chi inflammatory monocytes (region 2) in the GL113 or 1A8 treated mice. (B) Representative dot plots of CD11b+Ly6G+Ly6Cint neutrophils (region 1) and CD11b+Ly6GLy6Chi inflammatory monocytes in the GL113 or RB6-8C5 depleted mice (region 2). (C) Mean total number per ear of neutrophils (PMN) or inflammatory monocytes (Mo) in neutrophil depleted or control treated, infected mice, +/− 1 s.d., 4–6 ears per group pooled from 2 independent experiments. (D–E) Mean total number of DCs (CD11c+MHCII+) per ear (D) and the mean percentage of RFP+ DCs per ear (E) 1 d after infection, +/− 1 s.d., 8–10 ears per group, pooled from 3 independent experiments. (F) Representative dot plot of CD11c+MHCII+-gated RFP uninfected and RFP+ infected dermal DCs, 24 hr after infection. (G) Representative histogram plots of MPO stained, RFP (gray filled) and RFP+ (black line) DCs 24 hr after infection. (H) Mean percentage of RFP or RFP+ DCs staining positive for MPO, +/− 1 s.d., 4 ears per group, pooled from two independent experiments. (I) Representative histogram plots of MPO stained, RFP (gray filled) and RFP+ (black line) DCs. Mice were treated with control or neutrophil-depleting antibodies 1 d before i.d. challenge and the dermal cells were analyzed 14 days after infection.