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Chitin-binding domains of Escherichia coli ChiA mediate interactions with intestinal epithelial cells in mice with colitis

Abstract : Background and aims: Inducible chitinase 3-like-1 is expressed by intestinal epithelial cells (IECs) and adheres to bacteria under conditions of inflammation. We performed a structure-function analysis of the chitin-binding domains encoded by the chiA gene, which mediates the pathogenic effects of adherent invasive Escherichia coli (AIEC). Methods: We created AIEC (strain LF82) with deletion of chiA (LF82-Delta chiA) or that expressed chiA with specific mutations. We investigated the effects of infecting different IEC lines with these bacteria compared with nonpathogenic E coli; chitinase activities were measured using the colloidal chitin-azure method. Colitis was induced in C57/Bl6 mice by administration of dextran sodium sulfate, and mice were given 10(8) bacteria for 15 consecutive days by gavage. Stool/tissue samples were collected and analyzed. Results: LF82-Delta chiA had significantly less adhesion to IEC lines than LF82. Complementation of LF82-Delta chiA with the LF82 chiA gene, but not chiA from nonpathogenic (K12) E coli, increased adhesion. We identified 5 specific polymorphisms in the chitin-binding domain of LF82 chiA (at amino acids 362, 370, 378, 388, and 548) that differ from chiA of K12 and were required for LF82 to interact directly with IECs. This interaction was mediated by an N-glycosylated asparagine in chitinase 3-like-1 (amino acid 68) on IECs. Mice infected with LF82, or LF82-Delta chiA complemented with LF82 chiA, developed more severe colitis after administration of dextran sodium sulfate than mice infected with LF82-Delta chiA or LF82 that expressed mutant forms of chiA. Conclusions: AIEC adheres to an N-glycosylated chitinase 3-like-1 on IECs via the chitin-binding domain of chiA. This mechanism promotes the pathogenic effects of AIEC in mice with colitis.
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Submitted on : Friday, May 29, 2020 - 5:19:45 PM
Last modification on : Tuesday, February 23, 2021 - 11:42:03 AM

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Daren Low, Hoa T. Tran, In-Ah Lee, Nicolas Dreux, Alan Kamba, et al.. Chitin-binding domains of Escherichia coli ChiA mediate interactions with intestinal epithelial cells in mice with colitis. Gastroenterology, WB Saunders, 2013, 145 (3), pp.602-612. ⟨10.1053/j.gastro.2013.05.017⟩. ⟨hal-02651692⟩



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