Figures and data
CD131 contributed to dextran sulfate sodium (DSS)-induced murine colitis. (a) Gene expression dataset shows the change of relative Csf2rb gene expression in murine colon tissues with time after DSS administration. (b) Relative Csf2rb mRNA expression level in colon tissues of control and DSS- treated wt mice. (c-h) Comparison between wt and CD131-deficient mice during steady state (administrating normal drinking water) and DSS-induced colitis, including: (c) relative body weight change; (d) colon length; (e) a group of exemplary hematoxylin and eosin (H&E) histology sections of murine colon showing inflammatory infiltration and tissue destruction; (f) an exemplary graph showing the gating strategy for identifying CD11b+Ly6G+ neutrophils on multi-color flow cytometry and their relative cell number in the colonic tissues normalized to all viable cells; (g) relative Tnf and Il1b mRNA expression levels in colon tissues; and (h) relative Csf2rb mRNA expression level in colon tissues. (i) Relative Tnf and Il1b mRNA expression levels in blood leukocytes of control and DSS-treated wt mice. (j) Relative CD11b+Ly6G+ neutrophils in the blood of control and DSS-treated wt mice, normalized to all viable CD45+ leukocytes. ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns, not significant.
CD131 was associated with immune cell infiltration signature. (a) Immune cell infiltration analysis of the colon tissues of wt and CD131-deficient mice during steady state and DSS-induced colitis. **, p < 0.01; *, p < 0.05; ns, not significant. (b) Correlation analysis of the estimated infiltrating immune cells. (c) GSEA of colon tissues between wt and CD131-deficient mice showing that inflammatory bowel disease-related pathway was enriched in wt mice. (d) Venn graph showing 1411 DEGs between control and DSS-treated wt mice, as well as 17760 non-DEGs between control and DSS-treated CD131- deficient mice; the intersection resulted in 573 genes.
Macrophages and CD131 synergistically contributed to DSS-induced murine colitis. (a) Exemplary graphs showing the gating strategy for identifying CD11b+MHCII+F4/80+ macrophages on multi-color flow cytometry and their relative cell number, normalized to all viable cells, in the colonic tissues of wt and CD131-deficient mice treated with normal drinking water or DSS. (b) Exemplary graphs showing the gating strategy for identifying CD131+F4/80+ macrophages. (c-f) Comparison between wt and CD131-deficient mice treated with DSS plus either PBS liposomes or clodronate liposomes, including: (c) relative body weight at day 7; (d) colon length; (e) relative number of macrophages in the colonic tissues normalized to all viable cells; and (f) relative Tnf mRNA expression level in colon tissues. (g) Multi-color flow cytometry showing TNF mean fluorescence intensity (MFI) on colon macrophages in wt and CD131-deficient mice treated with normal drinking water or DSS. (h-i) RAW 264.7 murine macrophage cell line treated with control or rm-GM-CSF, and relative Tnf mRNA expression level (h) and CFSE MFI (i) were observed. ***, p < 0.001; **, p < 0.01; *, p < 0.05.
CCL4 mediated the effect of CD131 on chemotaxis and inflammatory response. (a-c) Comparison between wt and CD131-deficient mice treated with normal drinking water or DSS, including: (a) an exemplary graph showing the gating strategy for identifying CD11b+Ly6C+ monocytes on multi-color flow cytometry, and their relative cell number in the blood normalized to all viable CD45+ leukocytes; (b) an exemplary graph showing the gating strategy for identifying CD3+ T cells on multi-color flow cytometry, as well as their relative cell number in the colonic tissues normalized to all viable cells, and in the blood normalized to all viable CD45+ leukocytes; (c) relative Ccl4 mRNA expression in the colon tissues. (d-i) Comparison between wt mice treated with DSS plus IgG isotype control and anti-CCL4 antibodies, including: (d) relative macrophage number in the colon tissues normalized to all viable cells; (e) relative T cell number in the colon tissues normalized to all viable cells; (f) relative body weight at day 7; (g) colon length; (h) relative neutrophil number in the colon tissues normalized to all viable cells; and (i) relative Tnf mRNA expression in the colon tissues. ***, p < 0.001; **, p < 0.01; *, p < 0.05.
CD131 was associated with endoscopic and pathological severity of ulcerative colitis as well as macrophage and T cell infiltration into the colon. (a) Gene expression dataset shows relative CSF2RB gene expression in colonic tissues of healthy controls (HC), uninflamed ulcerative colitis (UC) patients and inflamed UC patients. **, p < 0.01. (b-e) a group of exemplary graphs showing histology sections of colonic tissues from UC patients stained with H&E (b), CD131 (c), CD68 (d) and CD3 (e). (f) Immunohistochemistry (IHC) analysis showing correlation between CD131 integrated density (IntDen), reflecting its expression level, and Robarts histopathological index (RHI) on H&E, reflecting disease severity. (g) IHC analysis showing correlation between CD131 IntDen and CD68 area, reflecting relative positive macrophage cell number. (h) IHC analysis showing correlation between CD131 IntDen and CD3 area, reflecting relative positive T cell number.
