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Quantification and Characterization of Mucosa-Associated and Intracellular Escherichia coli in Inflammatory Bowel Disease

Elliott, Timothy R. MBBS*,†,‡; Hudspith, Barry N. MSc*; Wu, Guanghui PhD§; Cooley, Michael PhD; Parkes, Gareth PhD*,†; Quiñones, Beatriz PhD; Randall, Luke PhD§; Mandrell, Robert E. PhD; Fagerquist, Clifton K. PhD; Brostoff, Jonathan MD*; Rayment, Neil B. MSc*; Boussioutas, Alex PhD; Petrovska, Liljana PhD§; Sanderson, Jeremy D. MD*,†

doi: 10.1097/MIB.0b013e3182a38a92
Original Basic Science Articles

Background: Mucosa-associated Escherichia coli are abundant in inflammatory bowel disease (IBD), but whether these bacteria gain intracellular access within the mucosa is uncertain. If E. coli does gain intracellular access, the contribution of bacterial pathogenicity to this requires further elucidation. This study aimed to quantify and characterize mucosa-associated and intracellular E. coli in patients with IBD and in healthy control subjects (HC).

Methods: Mucosal biopsies from 30 patients with Crohn's disease (CD), 15 with ulcerative colitis (UC), and 14 HC were cultured with or without gentamicin protection to recover intracellular or mucosa-associated E. coli, respectively. Overall, 40 strains (CD: n = 24, UC: n = 9, and HC: n = 7) were characterized by phylogenetic typing, adhesion and invasion assays, detection of virulence factors, antimicrobial resistance genes, and proteomic analysis.

Results: Mucosa-associated E. coli were more abundant in CD and UC than in HC (2750 versus 1350 versus 230 median colony-forming units per biopsy; P = 0.01). Intracellular E. coli were more prevalent in CD (90%) than in UC (47%) or HC mucosal biopsies (0%) (P < 0.001). Of 24 CD strains, 2 were adherent and invasive, but there were no unifying pathogenicity determinants that could distinguish most CD strains from UC or HC strains, or intracellular isolates from mucosa-associated isolates.

Conclusions: Intracellular E. coli are more common in CD than in UC and not identified in HC. Most intracellular E. coli did not have characterizing pathogenic features, suggesting a significant role for defects in mucosal immunity or barrier dysfunction in their ability to gain intracellular access.

Article first published online 28 August 2013

*Diabetes and Nutritional Sciences Division, King's College London, London, United Kingdom;

Department of Gastroenterology, Guy's and St. Thomas' NHS Foundation Trust, St Thomas' Hospital, London, United Kingdom;

Department of Medicine, University of Melbourne, Melbourne, Australia;

§Animal Health and Veterinary Laboratories Agency, Surrey, United Kingdom; and

US Department of Agriculture, Agricultural Research Service, Western Regional Research Center, Produce Safety and Microbiology Research Unit, Albany, California.

Reprints: Jeremy D. Sanderson, MD, Diabetes and Nutritional Sciences Division, King's College London, Franklin Wilkins Building, 150 Stamford St, London SE1 9NH, United Kingdom (e-mail: jeremy.sanderson@kcl.ac.uk).

The authors have no conflicts of interest to disclose.

Received February 22, 2013

Accepted March 13, 2013

© Crohn's & Colitis Foundation of America, Inc.
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