Increased infiltration of defective neutrophils in the kidneys of CD11cΔSyk mice.

<p>Control and CD11cΔSyk mice were infected with 2×10⁵ CFU of <i>C. albicans</i> intravenously. Kidneys were removed from naïve mice or mice 2 days post-infection and leukocytes were surface stained for CD11b, F4/80, MHC-II and Ly-6G. (A) Percentage and total number of Ly6G⁺ CD11b⁺ kidney neutrophils. Data are mean +/− SEM from four pooled independent experiments with each data point representing an individual mouse. (B) Representative expression of CD11b and CD11a on infiltrating kidney neutrophils of day 2 infected control and CD11cΔSyk mice. (C) Global geometric mean of CD11b expression on infiltrating kidney neutrophils normalized against control naïve mice. Data are mean +/− SEM from four independent experiments with each data point representing an individual mouse. (D) Neutrophils from day 2 infected mice were permeabilized and stained for MPO. Data shown are global geometric mean of MPO signal on infiltrating kidney neutrophils normalized against control mice. Data are combined from two independent experiments with each data point representing an individual mouse. (E) Control and CD11cΔSyk were infected with 2×10⁵ CFU of <i>C. albicans</i>-GFP intravenously. Kidneys were removed 2 days post-infection and analyzed for GFP expression. Representative staining profiles of total CD45.2⁺ kidney cells (top panel) and CD45.2⁺ Ly-6G⁺ neutrophils (bottom panel). Boxes indicate the percentages of GFP⁺ neutrophils and the mean +/− SEM from three independent experiments with number (n) of mice indicated. (F) GFP⁺ and GFP⁻ neutrophils (CD11b⁺ Ly-6G⁺ CD11c⁻ F4/80⁻) were sorted, lysed in water and plated to determining presence of viable fungi. Data are combined from two independent experiments with each data point representing cells sorted from an individual mouse. (G) Neutrophils were sorted from naïve BM or day 2 infected kidneys of control and CD11cΔSyk mice and incubated with <i>C. albicans</i> (10∶1) for 1 h at 37°C. The survival of fungi was then assessed. Data are mean +/− SEM from three independent experiments with each data point representing cells sorted from an individual mouse. Statistical significance of any differences for A, C, D and G was determined by 2-tailed <i>t</i> test. Whilst a Kruskal-Wallis with Dunn's multiple comparison test was undertaken for F. NS, not significant.