Figure 6: IL-10 competent macrophages can rescue survival after infection.

Il10^−/− Rag^−/− mice were infected with C. rodentium. On day 1 after infection, macrophages (CD11b⁺CD11c^intF4/80⁺ cells) were sorted from large intestine of Il10^gfp reporter mice and 5 × 10⁵ cells per mouse of GFP⁺ (IL-10⁺) or GFP⁻ (IL10⁻) populations were injected intravenously. (a) Gating strategy for obtaining GFP⁺ (IL-10⁺) and GFP⁻ (IL-10⁻) colonic macrophages for adoptive cell transfer. CD45⁺ colonic lamina propria cells were selected on the basis of CD11b⁺, CD11c^int expression (gate #1) and F4/80 expression (gate #2) and further separated into GFP⁺(IL-10⁺) (gate #4) or GFP⁻ (IL-10⁻; gate #3) populations. (b) Survival and histology score at day 6 after infection were monitored and is presented as a summary of two independent experiments with 3–5 mice per group. (c) (Top) total cell numbers were counted from large intestine at day 6 after infection. Frequencies of monocytes (Ly6G⁻Ly6C^high) and granulocytes (Ly6G^highLy6C^low) were measured by flow cytometry and cell numbers were calculated according to the respective total cell number. Cell numbers are presented as averages and s.d. from three mice per group from two independent experiments. (Bottom) analysis of Ccl3, Ccl4 and Ccl5 mRNA transcripts in the large intestine at day 6 after infection. Data are presented as averages with s.d. from three mice per group from two independent experiments. Student’s t-test, **P<0.01, ***P<0.001.