Clinical Research
Copyright ©2008 The WJG Press and Baishideng. All rights reserved.
World J Gastroenterol. Dec 28, 2008; 14(48): 7345-7352
Published online Dec 28, 2008. doi: 10.3748/wjg.14.7345
Disruption of colonic barrier function and induction of mediator release by strains of Campylobacter jejuni that invade epithelial cells
Johannes Beltinger, Jo del Buono, Maeve M Skelly, John Thornley, Robin C Spiller, William A Stack, Christopher J Hawkey
Johannes Beltinger, Jo del Buono, Maeve M Skelly, John Thornley, Robin C Spiller, William A Stack, Christopher J Hawkey, Wolfson Digestive Diseases Centre, Nottingham University Hospital, QMC Campus, Nottingham NG7 2UH, United Kingdom
Author contributions: Stack WA and Hawkey CJ conceived and co-ordinated the study and raised funding; Beltinger J did the majority of the work, ably assisted by del Buono J and Skelly MM; Thornley J and Spiller RC kindly provided samples; All authors contributed to the design conduct and authorship of the paper which was originally drafted by Beltinger J.
Supported by The Medical Research Council (UK), No. G9716348
Correspondence to: Christopher J Hawkey, Wolfson Digestive Diseases Centre, Nottingham University Hospital, QMC Campus, Nottingham NG7 2UH, United Kingdom. cj.hawkey@nottingham.ac.uk
Telephone: +44-115-9709918 Fax: +44-115-9422232
Received: July 9, 2008
Revised: October 28, 2008
Accepted: November 4, 2008
Published online: December 28, 2008
Abstract

AIM: To study the mechanisms by which Campylobacter jejuni (C. jejuni) causes inflammation and diarrhea. In particular, direct interactions with intestinal epithelial cells and effects on barrier function are poorly under-stood.

METHODS: To model the initial pathogenic effects of C. jejuni on intestinal epithelium, polarized human colonic HCA-7 monolayers were grown on permeabilized filters and infected apically with clinical isolates of C. jejuni. Integrity of the monolayer was monitored by changes in monolayer resistance, release of lactate dehydrogenase, mannitol fluxes and electron microscopy. Invasion of HCA-7 cells was assessed by a modified gentamicin protection assay, translocation by counting colony forming units in the basal chamber, stimulation of mediator release by immunoassays and secretory responses in monolayers stimulated by bradykinin in an Ussing chamber.

RESULTS: All strains translocated across monolayers but only a minority invaded HCA-7 cells. Strains that invaded HCA-7 cells destroyed monolayer resistance over 6 h, accompanied by increased release of lactate dehydrogenase, a four-fold increase in permeability to [3H] mannitol, and ultrastructural disruption of tight junctions, with rounding and lifting of cells off the filter membrane. Synthesis of interleukin (IL)-8 and prostaglandin E2 was increased with strains that invaded the monolayer but not with those that did not.

CONCLUSION: These data demonstrate two distinct effects of C. jejuni on colonic epithelial cells and provide an informative model for further investigation of initial host cell responses to C. jejuni.

Keywords: Campylobacter jejuni; Cell invasion; Cell culture; Chloride secretion; Colonocyte; HCA-7 cells; Membrane permeability; Monolayer; Mucosal barrier; Ussing chamber