Enterococcus faecalis metalloprotease compromises epithelial barrier and contributes to intestinal inflammation

Gastroenterology. 2011 Sep;141(3):959-71. doi: 10.1053/j.gastro.2011.05.035. Epub 2011 May 26.

Abstract

Background & aims: Matrix metalloproteases (MMPs) mediate pathogenesis of chronic intestinal inflammation. We characterized the role of the gelatinase (GelE), a metalloprotease from Enterococcus faecalis, in the development of colitis in mice.

Methods: Germ-free, interleukin-10-deficient (IL-10(-/-)) mice were monoassociated with the colitogenic E faecalis strain OG1RF and isogenic, GelE-mutant strains. Barrier function was determined by measuring E-cadherin expression, transepithelial electrical resistance (TER), and translocation of permeability markers in colonic epithelial cells and colon segments from IL-10(-/-) and TNF(ΔARE/Wt) mice. GelE specificity was shown with the MMP inhibitor marimastat.

Results: Histologic analysis (score 0-4) of E faecalis monoassociated IL-10(-/-) mice revealed a significant reduction in colonic tissue inflammation in the absence of bacteria-derived GelE. We identified cleavage sites for GelE in the sequence of recombinant mouse E-cadherin, indicating that it might be degraded by GelE. Experiments with Ussing chambers and purified GelE revealed the loss of barrier function and extracellular E-cadherin in mice susceptible to intestinal inflammation (IL-10(-/-) and TNF(ΔARE/Wt) mice) before inflammation developed. Colonic epithelial cells had reduced TER and increased translocation of permeability markers after stimulation with GelE from OG1RF or strains of E faecalis isolated from patients with Crohn's disease and ulcerative colitis.

Conclusions: The metalloprotease GelE, produced by commensal strains of E faecalis, contributes to development of chronic intestinal inflammation in mice that are susceptible to intestinal inflammation (IL-10(-/-) and TNF(ΔARE/Wt) mice) by impairing epithelial barrier integrity.

Publication types

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

MeSH terms

  • Animals
  • CD4-Positive T-Lymphocytes / physiology
  • Cadherins / metabolism
  • Cell Membrane Permeability / physiology*
  • Colitis / etiology
  • Colitis / metabolism
  • Colitis / physiopathology*
  • Disease Models, Animal
  • Enterococcus faecalis / metabolism*
  • Gelatinases / metabolism*
  • Gram-Positive Bacterial Infections / complications
  • Gram-Positive Bacterial Infections / metabolism
  • Gram-Positive Bacterial Infections / physiopathology*
  • Interleukin-10 / genetics
  • Interleukin-10 / metabolism
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / metabolism*
  • Metalloproteases / metabolism*
  • Mice
  • Mice, Knockout
  • Mice, Mutant Strains
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Cadherins
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • Metalloproteases
  • Gelatinases