Interleukin-10 blocked endoplasmic reticulum stress in intestinal epithelial cells: impact on chronic inflammation

Gastroenterology. 2007 Jan;132(1):190-207. doi: 10.1053/j.gastro.2006.10.030. Epub 2006 Oct 21.

Abstract

Background & aims: The initiation of endoplasmic reticulum (ER)-mediated stress responses in intestinal epithelial cells (IEC) may contribute to the pathogenesis of chronic intestinal inflammation. The aim of the study was to use functional epithelial cell proteomics to characterize anti-inflammatory mechanisms of interleukin 10 (IL-10).

Methods: Primary IEC were isolated from Enterococcus faecalis-monoassociated IL-10-deficient (IL-10-/-) and wild-type mice to perform 2D-gel sodium-dodecyl-sulfate polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. In addition, IEC from 6 patients with active Crohn's disease, ulcerative colitis, and sigmoid diverticulitis as well as noninflamed controls were purified. Molecular protective mechanisms of IL-10 were characterized in tumor necrosis factor (TNF)-stimulated IL-10 receptor (IL-10R) reconstituted epithelial cells.

Results: Primary IEC from IL-10-/- mice as well as inflammatory bowel disease patients revealed increased expression levels of the glucose-regulated ER stress protein (grp)-78 under conditions of chronic inflammation. Consistent with the observation that TNF induced ER stress responses through grp-78 redistribution from the ER lumen to the cytoplasmic IkappaB kinase complex, grp-78 knockdown completely abolished TNF-induced nuclear factor-kappaB RelA phosphorylation in epithelial cell cultures. Interestingly, IL-10 inhibited grp-78 protein and messenger RNA expression in IL-10R reconstituted IEC. Chromatin immunoprecipitation analysis and immunofluorescence microscopy revealed that IL-10-mediated p38 signaling inhibited TNF-induced recruitment of the ER-derived activating transcription factor (ATF)-6 to the grp-78 promoter likely through the blockade of ATF-6 nuclear translocation.

Conclusions: Primary IEC from inflamed IL-10-/- mice and inflammatory bowel disease patients revealed activated ER stress responses in the intestinal epithelium. IL-10 inhibits inflammation-induced ER stress response mechanisms by modulating ATF-6 nuclear recruitment to the grp-78 gene promoter.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activating Transcription Factor 6 / metabolism
  • Animals
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Chronic Disease
  • Endoplasmic Reticulum / immunology
  • Endoplasmic Reticulum / metabolism*
  • Enterococcus faecalis
  • Epithelial Cells / cytology
  • Epithelial Cells / immunology
  • Gram-Positive Bacterial Infections / immunology
  • Gram-Positive Bacterial Infections / metabolism
  • Heat-Shock Proteins / genetics
  • Heat-Shock Proteins / metabolism
  • Humans
  • Inflammatory Bowel Diseases / immunology*
  • Inflammatory Bowel Diseases / metabolism
  • Inflammatory Bowel Diseases / microbiology
  • Interleukin-10 / genetics
  • Interleukin-10 / immunology*
  • Interleukin-10 / metabolism
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / immunology*
  • Intestinal Mucosa / pathology
  • Mice
  • Mice, Inbred Strains
  • Mice, Mutant Strains
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism
  • Promoter Regions, Genetic / immunology
  • Proteomics
  • Signal Transduction / immunology
  • Transcription Factor RelA / metabolism
  • Tumor Necrosis Factor-alpha / metabolism
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Activating Transcription Factor 6
  • Atf6 protein, mouse
  • Heat-Shock Proteins
  • Molecular Chaperones
  • Rela protein, mouse
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • p38 Mitogen-Activated Protein Kinases
  • molecular chaperone GRP78