Panel A shows a curve plot with data points of body weight variation over days post-infection under different cohousing conditions, where the x-axis represents days post-infection and the y-axis represents body weight variation in percent. Panel B shows a curve plot with data points of fecal Citrobacter rodentium burden over days post-infection, where the x-axis represents days post-infection and the y-axis represents colony forming units per gram. Panel C shows a scatter plot of colon length at 10 days post-infection, where the y-axis represents colon length in centimeters. Panel D shows a scatter bar plot of fecal lipocalin-2 levels at 10 days post-infection, where the y-axis represents lipocalin-2 in nanograms per milliliter. Panel E shows a scatter bar plot of intestinal permeability measured by fluorescein isothiocyanate dextran at 10 days post-infection, where the y-axis represents fluorescein isothiocyanate dextran in micrograms per milliliter. Panel F shows representative hematoxylin and eosin stained sections of small intestine and colon along with scatter bar plots of histopathology scores, where the y-axis represents histopathological score. Panel G shows flow cytometry plots and scatter bar plots of myeloid cell populations in the colon at 10 days post-infection, where the y-axis represents number of cells. Panel H and I show flow cytometry plots and scatter bar plots of small intestine ILC3s and cytokine producing subsets after ex vivo interleukin 23 stimulation, where the y-axis represents number of ILC3s or percentage of cytokine positive cells. Panel J and K show flow cytometry plots and scatter bar plots of colonic ILC3s and cytokine producing subsets after ex vivo interleukin 23 stimulation, where the y-axis represents number of ILC3s or percentage of cytokine positive cells. Panel L shows a schematic of interleukin 22 neutralization or recombinant interleukin 22 treatment during infection. Panel M shows a curve plot with data points of body weight variation over days post-infection under different treatment conditions, where the x-axis represents days post-infection and the y-axis represents body weight variation in percent. Panel N shows a scatter plot of fecal Citrobacter rodentium burden at 10 days post-infection, where the y-axis represents colony forming units per gram.
Lingo4 −/− mice are more susceptible to C. rodentium infection due to impaired IL-22–mediated mucosal protection. (A) Body weight variation in cohoused and non-cohoused WT and Lingo4−/− mice during C. rodentium infection. Mean ± SEM, n = 15; three experiments combined. (B) Fecal C. rodentium burden at 5, 10, 15, and 20 d.p.i. Mean ± SEM, n = 9–11; two experiments combined. (C–E) Colon length (C), fecal lipocalin-2 levels (D), and intestinal permeability by FITC-dextran (E) at 10 d.p.i. Mean ± SEM, n = 9–11 (C–D) and n = 5 (E). (F) Representative H&E sections of colon and small intestine with histopathological scoring at 10 d.p.i. Mean ± SEM, n = 5. Scale bars = 100 μm. (G) Representative plots and absolute numbers of myeloid cells in colon at 10 d.p.i. Mean ± SEM, n = 5; representative of two experiments. “NO,” neutrophils; “Mono,” monocytes; “Mac,” macrophages; “DC,” dendritic cells. (H and I) Representative plots and absolute numbers of siLP ILC3s (H) and IL-22+, IL-17+, and IFN-γ+ siLP ILC3 (I) frequencies after ex vivo IL-23 stimulation at 10 d.p.i. Mean ± SEM, n = 5; representative of two experiments. ILC3s were defined as live lymphocyte-sized Lin– (CD3− CD19−) Thy1.2hi CD45int cells. (J and K) Representative plots and absolute numbers of colonic ILC3s (J) and IL-22+, IL-17+, and IFN-γ+ colonic ILC3 (K) frequencies after ex vivo IL-23 stimulation at 10 d.p.i. Mean ± SEM, n = 5; representative of two experiments. ILC3s were defined as live lymphocyte-sized Lin– (CD3– CD19–) Thy1.2hi CD45int cells. (L–N) Schematic of IL-22 neutralization or recombinant IL-22 (rmIL-22) therapy during infection (L), relative body weight (M), and fecal C. rodentium burden at 10 d.p.i. (N). Mean ± SEM, n = 4. (A–N) Statistical significance was determined using unpaired two-tailed t test or one-way ANOVA with Tukey’s post hoc test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.