IL-10 Promotes Production of Intestinal Mucus by Suppressing Protein Misfolding and Endoplasmic Reticulum Stress in Goblet Cells

doi:10.1053/j.gastro.2012.10.043

Gastroenterology

Volume 144, Issue 2, February 2013, Pages 357-368.e9

https://doi.org/10.1053/j.gastro.2012.10.043 Get rights and content

Background & Aims

Protein misfolding and endoplasmic reticulum (ER) stress have been observed in intestinal secretory cells from patients with inflammatory bowel diseases and induce intestinal inflammation in mice. However, it is not clear how immune factors affect ER stress and therefore disease symptoms.

Methods

We analyzed the effects of interleukin (IL)-10 on ER stress in intestinal tissues in wild-type C57BL/6, Winnie, IL-10^−/−, and Winnie × IL-10^+/− mice. In Winnie mice, misfolding of the intestinal mucin Muc2 initiates ER stress and inflammation. We also analyzed the effects of different inhibitors of IL-10 signaling and the N-glycosylation inhibitor tunicamycin in cultured human LS174T goblet cells.

Results

Administration of neutralizing antibodies against IL-10 or its receptor (IL-10R1) to Winnie mice rapidly exacerbated ER stress and intestinal inflammation compared with mice given vehicle (controls). Antibodies against IL-10 also increased accumulation of misfolded Muc2 in the ER of goblet cells of Winnie mice and increased T-cell production of inflammatory cytokines. Winnie × IL-10^+/− mice and IL-10^−/− mice with a single Winnie allele each developed more severe inflammation than Winnie mice or IL-10^−/− mice. Administration of tunicamycin to wild-type mice caused intestinal ER stress, which increased when IL-10R1 was blocked. In LS174T cells, induction of ER stress with tunicamycin and misfolding of MUC2 were reduced by administration of IL-10; this reduction required STAT1 and STAT3. In LS174T cells incubated with tunicamycin, IL-10 up-regulated genes involved in MUC2 folding and in ER-associated degradation and maintained correct folding of MUC2, its transport from the ER, and its O-glycosylation and secretion.

Conclusions

IL-10 prevents protein misfolding and ER stress by maintaining mucin production in goblet cells and helps the intestine preserve the mucus barrier.

Section snippets

Materials and Methods

See Supplementary Materials and Methods for additional information.

Deficiency in IL-10 Combines With Intestinal ER Stress to Cause Severe Colitis

To determine the importance of IL-10 in suppression of ER stress–initiated colitis, we crossed Winnie (missense misfolding mutation in Muc2) and IL-10^−/− mice. As expected,7, 21 both IL-10^−/− and Winnie mice developed mild colitis, and mice heterozygous for either the IL-10 null allele or Muc2^Win allele did not develop colitis. However, strikingly, Winnie mice haplosufficient for IL-10 and IL-10^−/− mice carrying a single Muc2^Win allele developed very severe inflammation through all regions of

Discussion

Protein accumulation within the ER results in the activation of UPR pathways that control ER stress and protect the cell from damage but can induce inflammatory signaling.3, 29, 30 A growing body of evidence suggests that ER stress in secretory cells contributes to pathology in IBD, and therefore modulating ER stress has therapeutic potential.4, 6, 31 In the intestinal microenvironment, IL-10 is produced by regulatory T cells, T effector cells, macrophages, dendritic cells, and epithelial

Acknowledgements

The authors thank all of the members of the Mater Medical Research Institute for their help and support.

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: Conflicts of interest The authors disclose no conflicts.

: Funding Supported by National Health and Medical Research Council project grant 604304. M.A.M. is supported by a National Health and Medical Research Council Senior Research Fellowship. T.H.F. is supported by a National Health and Medical Research Council Practitioner Fellowship.

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Original ResearchBasic and Translational—Alimentary TractIL-10 Promotes Production of Intestinal Mucus by Suppressing Protein Misfolding and Endoplasmic Reticulum Stress in Goblet Cells