Figure 4

IL-36-deficient mice display altered innate mucosal inflammation and bacterial colonization in C. rodentium-induced colitis. (a) Representative bioluminescence images of (il36r+/−) littermate ctrl and il36r−/− mice at day 7 after PBS treatment of infection with C. rodentium obtained using the Xenogen—IVIS Lumina II and showing (a) liver; (b) cecum; (c) colon; (d) spleen; and (e) mesenteric lymph nodes of each experimental group. Color scales represent bioluminescence signal intensity (p s⁻¹ cm⁻² per sr) with red representing the most intense light emission and blue the weakest signal. (b) Colonic bioluminescent signal total flux (p/s) was quantified using the same ROIs for all infected animals and including all the distal colons, background was corrected for all animals. Data show total flux (p/s) on the distal colons of ctrl and il36r−/− infected with C. rodentium on days 7 and 14 p.i. and is expressed as mean±s.e.m. (n= 5–7 mice per group). Significance was determined using one-way ANOVA and Bonferroni post hoc **P=0.01. Expression levels of (c) mKC and (d) MPO activity in colonic homogenates of uninfected (PBS) or C. rodentium-infected ctrl and il36r−/− mice at day 7 p.i. Protein expression/activity is expressed as pg mg⁻¹ of total tissue protein from five individual mice per group. Relative percentages of (e) total macrophage (CD11b⁺F480⁺) and (f) inflammatory macrophage (CD11b⁺F480⁺Gr1⁺) subsets in the mesenteric lymph nodes of ctrl and il36r−/− infected with C. rodentium on day 7 p.i. Each symbol represents values for an individual mouse with five mice per group. (g) Representative fluorescence activated cell sorting showing values from individual mice in (f). Data expressed as mean±s.e.m. with significance determined by unpaired student’s t-test, *P≤0.05 and **P≤0.01. ANOVA, analysis of variance; MPO, myeloperoxidase; PBS, phosphate-buffered saline; p.i., post infection; ROIs, regions of interest.

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