1. IL-10R-deficient macrophages secrete IL-23, inducing IL-22 secretion by ILC3 and TH17 cells.
IL-10R-deficient macrophages secrete IL-23, inducing IL-22 secretion by ILC3 and TH17 cells. (A) Volcano plot of statistical significance (−log10P value) against log2 ratio of macrophages sorted from the colonic lamina propria of 6- to 7-week-old Cx3cr1creIl10rafl/fl and Cx3cr1gfp/+ mice, based on RNA-seq data. Significantly up- or down-regulated genes (fold change, >2; adj. P < 0.05) are in black, and relevant proinflammatory up-regulated genes are highlighted in red. Data are representative of two independent experiments, n ≥ 3 for each group. (B) RNA-seq–normalized read numbers for single genes of interest are plotted separately; each dot represents one mouse. (C) qRT-PCR analysis of Il23p19 expression by sorted colonic macrophages from 6- to 7-week-old mice (left) or of Il22 expression (right) in colonic whole-tissue extracts of 6- to 7-week-old indicated mice, including antibiotics (Abx) treatment. Data were collected from two independent experiments, n ≥ 3 in each group. (D) qRT-PCR analysis of Il17 expression in colonic whole-tissue extracts of indicated mice. Data were collected from two independent experiments, n ≥ 3 in each group. (E) Whole-mount staining of colonic tissue of Cx3cr1cre:Il10rafl/fl:Rorgtgfp mice and Cre-negative littermates (age, 3 to 4 months), analyzed by confocal microscope, GFP in green, and CD3 in red. Indent in the bottom is in the white rectangle. Scale bars, 50 μm (top) and 20 μm (bottom). (F) Representative picture of flow cytometry analysis and sorting strategy of Cx3cr1cre:Il10rafl/fl:Rorgtgfp mice and Cre-negative littermates 3 to 4 months old. (G and H) qRT-PCR analysis of RNA extracted from sorted colonic TH17 cells (F) or ILC3s (G) for indicated genes. Data were collected from two independent experiments, n = 3 in each group. *P ≤ 0.05.
Biana Bernshtein et al. Sci. Immunol. 2019;4:eaau6571
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