Macrophage dysfunction initiates colitis during weaning of infant mice lacking the interleukin-10 receptor
- Boston Children's Hospital, United States
- Harvard Medical School, United States
- Massachusetts Institute of Technology, United States
- Brigham and Women's Hospital, United States
- Edmond and Lily Safra Children’s Hospital, Sheba Medical Center, Israel
- Tel Aviv University, Israel
- University of Manchester, United Kingdom
- Hannover Medical School, Germany
- Brigham and Women’s Hospital, United States
Figures
Figure 1 with 1 supplement
Spontaneous colitis develops in Il10rb-/- mice between 3–4 weeks.
(A) Representative histologic images of distal colon from Il10rb-/- and control mice at indicated age (10X magnification for 8 week and 30 week Il10rb-/-, scale = 200 μm; and 20X for all other …
Figure 1—figure supplement 1
Severe colitis in 6 month old Il10rb-/- mice.
(A) Representative endoscopic images, (B) Endoscopy score, n = 10–12; (C) Representative gross colon images, and (D) Colon tissue weights without fecal contents at 6 months (30 weeks). Each data …
Figure 2 with 2 supplements
Colonic lamina propria macrophage dysfunction in 3–4 week old Il10rb-/- mice.
Representative FACS plots of (A) MΦ subsets, gated on CD45+CD11b+ CD11cintCD103-CD64+ cells in the LP of Il10rb-/- and littermate Il10rb+/- mice at 2 and 3–4 weeks. Gating strategy is shown in the …
Figure 2—figure supplement 1
Gating strategy for colonic lamina propria macrophages.
https://doi.org/10.7554/eLife.27652.006
Figure 2—figure supplement 2
Minimal alterations in colonic lymphocytes and dendritic cells occur in the absence of IL10Rβ signaling in 3–4 week old mice.
Representative FACS plots of (A) FoxP3+ Tregs, gated on CD45+CD3e+CD4+ T cells, (B) CD4+ Th effectors following PMA and ionomycin stimulation for 4.5 hr, gated on CD45+CD3e+CD4+ T cells, (C) Type …
Figure 3
Lamina propria macrophages of infant Il10rb-/- mice exhibit proinflammatory transcriptional profile.
Colonic LP MΦs (P3/P4; gated on CD45+CD11b+CD11cint CD103-CD64+Ly6C-MHCII+ cells) were sorted (minimum 10,000 cells), RNA was prepared, and RNA-seq was performed. (A) Heatmap showing relative …
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Figure 3—source data 1
R-script used to generate differential gene expression data shown in Figure 3A.
- https://doi.org/10.7554/eLife.27652.009
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Figure 3—source data 2
R-script used to generate heat map shown in Figure 3A.
- https://doi.org/10.7554/eLife.27652.010
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Figure 3—source data 3
Normalized gene expression values used to construct heat map shown in Figure 3A.
- https://doi.org/10.7554/eLife.27652.011
Figure 4
Liposomal clodronate-mediated depletion of colonic macrophages ameliorates colitis in infant Il10rb-/- mice.
Three week old Il10rb-/- mice were injected intraperitoneally (i.p.) with liposomal control or liposomal-clodronate for three times within a week. (A) Representative histologic images of distal …
Figure 5
IL10Rα is necessary to prevent intestinal inflammation in infant mice.
Il10ra-/- mice (C57BL/6) were crossed with congenic Cdcs1 mice and the development of spontaneous colitis was investigated. (A) Representative histologic images from the colon of Cdcs1+/+Il10ra-/- …
Figure 6 with 1 supplement
Macrophage-specific IL10Rα is necessary to prevent intestinal inflammation in infant mice.
(A) Representative gross macroscopic images of colon from 6 week old mice. (B) Representative histologic images of distal colons of Cdcs1+/+Il10rafl/flLyz2Cre and littermate Cdcs1+/+Il10rafl/fl …
Figure 6—figure supplement 1
Immunophenotyping of 3–4 week old Cdcs1+/+Il10rafl/fl Lyz2Cre and littermate Cdcs1+/+Il10rafl/fl control mice.
Representative FACS plots (A, B), and graphical comparison of frequencies and total numbers of intracellular cytokine positive CD4+ Th cells (C), and FoxP3+ Tregs (D) in the colons of 3–4 week old …
Figure 7
Antibiotics exposure prevents colitis in infant Il10rb-/- mice.
(A) Representative histological images (4X magnification in upper panel, scale = 500 μm; 10X in lower panel, scale = 200 μm for Il10rb+/-, Il10rb-/- Water, and Poly B; and 20X in lower panel, scale =…
Figure 8
IL10Rβ does not regulate fecal microbiome in developing mice.
Fecal pellets were collected aseptically every other day until day 28 and then weekly until day 98 from Il10rb-/- and littermate Il10rb+/- mice co-housed under SPF conditions. Composition of the …
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Figure 8—source data 1
Raw values of alpha diversity (Chao1), and quantification of OTU007 shown in Figure 8A and C, respectively.
- https://doi.org/10.7554/eLife.27652.018
Author response image 1
CCR2 blocking does not inhibit inflammatory gene expression in the infant Il10rb-/- mice.
(A) Flow cytometry analyses of colonic lamina propria macrophages of indicated mice after 5 days of treatment (20μg/mouse/day, i.p.) with anti-CCR2 (MC21) or rat IgG2b, κ (isotype). (B) Graphical …
