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Dual-association of gnotobiotic Il-10−/− mice with 2 nonpathogenic commensal bacteria induces aggressive pancolitis

Authors

  • Sandra C. Kim MD,

    Corresponding author
    1. Center for Gastrointestinal Biology and Disease, University of North Carolina, Chapel Hill, NC
    • Department of Pediatrics, Division of Gastroenterology, and the Center for Gastrointestinal Biology and Disease, CB #7299, Rm. 5143 Bioinformatics Building, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7299
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  • Susan L. Tonkonogy PhD,

    1. North Carolina State University College of Veterinary Medicine, Raleigh, NC
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  • Thomas Karrasch MD,

    1. Center for Gastrointestinal Biology and Disease, University of North Carolina, Chapel Hill, NC
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  • Christian Jobin PhD,

    1. Center for Gastrointestinal Biology and Disease, University of North Carolina, Chapel Hill, NC
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  • R. Balfour Sartor MD

    1. Center for Gastrointestinal Biology and Disease, University of North Carolina, Chapel Hill, NC
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Abstract

Background: Monoassociating gnotobiotic IL-10-deficient (−/−) mice with either nonpathogenic Enterococcus faecalis or a nonpathogenic Escherichia coli strain induces T-cell-mediated colitis with different kinetics and anatomical location (E. faecalis: late onset, distal colonic; E. coli: early onset, cecal). Hypothesis: E. faecalis and E. coli act in an additive manner to induce more aggressive colitis than disease induced by each bacterial species independently.

Methods: Germ-free (GF) inbred 129S6/SvEv IL-10−/− and wildtype (WT) mice inoculated with nonpathogenic E. faecalis and/or E. coli were killed 3–7 weeks later. Colonic segments were scored histologically for inflammation (0 to 4) or incubated in media overnight to measure spontaneous IL-12/IL-23p40 secretion. Bacterial species were quantified by serial dilution and plated on culture media. Mesenteric lymph node (MLN) CD4+ cells were stimulated with antigen-presenting cells pulsed with bacterial lysate (E. faecalis, E. coli, Bacteroides vulgatus) or KLH (unrelated antigen control). IFN-γ and IL-17 levels were measured in the supernatants.

Results: Dual-associated IL-10−/− (but not WT) mice developed mild-to-moderate pancolitis by 3 weeks that progressed to severe distal colonic-predominant pancolitis with reactive atypia and duodenal inflammation by 7 weeks. NF-κB was activated in the duodenum and colon in dual-associated IL-10−/− × NF-κBEGFP mice. The aggressiveness of intestinal inflammation and the degree of antigen-specific CD4+ cell activation were greater in dual- versus monoassociated IL-10−/− mice.

Conclusion: Two commensal bacteria that individually induce phenotypically distinct colitis in gnotobiotic IL-10−/− mice act additively to induce aggressive pancolitis and duodenal inflammation.

(Inflamm Bowel Dis 2007)

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