Effects of neutrophil depletion on the activation and function of dermal DCs <i>ex vivo</i>, and on CD4⁺ T cell priming <i>in vivo</i>.

<p>(A) Representative dot plots of RFP<b>⁺</b> DCs recovered 3 d after i.d. infection with 2×10⁶<i>Lm</i>-RFP metacyclic promastigotes in mice treated with GL113 or RB6-8C5 1 d before challenge. (B) Representative histogram plots of RFP⁺CD11c⁺MHCII⁺ gated cells from control treated (gray filled) and RB6-8C5 treated (black line) infected mice stained for MHCII, CD86, CD40 and CD80. (C) Mean fluorescence intensity (MFI) of MHCII, CD86, CD40 and CD80 expression on RFP⁺CD11c⁺MHCII⁺ gated cells from control treated (white bars) and RB6-8C5 treated (black bars) infected mice; mean percentage +/− 1 s.d. calculated from 2 independent experiments; * <i>p</i> = 0.0214, ** <i>p</i> = 0.0134, *** <i>p</i> = 0.0025. (D) Representative dot plots showing the gates used for sorting of the RFP<b>⁺</b>CD11c⁺ dermal cells recovered from control and RB6-8C5 treated, infected mice. (E) IFN-γ levels in supernatants of T cells from healed, <i>L. major</i>-infected mice co-cultured for 3 d with sorted RFP⁺CD11c⁺ cells recovered from the ears of control treated or RB6-8C5 treated mice 3 days after i.d. infection with 2×10⁶<i>Lm</i>-RFP metacyclic promastigotes. Two independent experiments are shown. (F–H) B6.SJL mice were treated with GL113, RB6-8C5 or 1A8 mAb followed 1 d later by i.d. inoculation with 10⁵<i>Lm</i> SP-OVA or <i>Lm</i> 3′NT metacyclic promastigotes. CFSE-labeled CD4⁺ OT-II cells were adoptively transferred at the time of (F–G) or 2 weeks after infection (F). Shown are representative dot and histogram plots of IFN-γ expression and/or CFSE fluorescence of CD45.2<b>⁺</b>CD4<b>⁺</b> gated cells from the draining lymph nodes 6 days after transfer, and the proliferative response with means of CD4⁺ OT-II cells from individual mice, as defined by the percent of cells with reduced CFSE content. The data are pooled from two independent experiments.