Fig. 4.

Inhibition of complement C5aR decreases cutaneous expression of innate immune and inflammatory mediators and skin inflammatory cell infiltration. (A) After 2-wk treatment with C5aRA or iC5aRA, skin was collected and RNA extracted for gene expression analysis. Skin mRNA levels were determined by quantitative real-time PCR (normalized to B2m) and expressed as fold change in C5aRA-treated transcript levels relative to iC5aRA-treated transcript levels, which were assigned an average value of 1. Data are means ± SD (n = 9 C5aRA-treated mice and n = 8 iC5aRA-treated mice from two independent experiments; *P < 0.05 and **P < 0.01). Table S2 shows all genes assayed and primer/probe sets. (B and C) Skin sections (6 µM thick) of mice treated with C5aRA and iC5aRA for 2 wk were stained with antibodies specific for (B) CD3 and (C) F4/80 to identify infiltrating lymphocytes and macrophages, respectively. Depicted are representative images at magnifications of 300× (Left) and the same image at a magnification of 600× (Right). Fig. S2 shows control and H&E staining. (D) For all staining, three to five fields per section were analyzed at magnification of 400× and positive cells were counted. Data are expressed as average number of immunoreactive cells in the field and are representative of one experiment consisting of three mice for each treatment and two skin biopsies per mouse. Error bars represent SEM (*P < 0.05). (Scale bars: Left, 100 µm; Right, 200 µm.)