title: Disruption of colonic barrier function and induction of mediator release by strains of Campylobacter jejuni that invade epithelial cells.
abstract: OBJECTIVE
  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 [(3)H] mannitol, and ultrastructural disruption of tight junctions, with rounding and lifting of cells off the filter membrane. Synthesis of interleukin (IL)-8 and prostaglandin E(2) was increased with strains that invaded the monolayer but not with those that did not.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Wolfson Digestive Diseases Centre, Nottingham University Hospital, QMC Campus, Nottingham NG7 2UH, United Kingdom.
authors.forename: Johannes
authors.initials: J
authors.lastname: Beltinger
authors.forename: Jo
authors.initials: J
authors.lastname: del Buono
authors.forename: Maeve M
authors.initials: MM
authors.lastname: Skelly
authors.forename: John
authors.initials: J
authors.lastname: Thornley
authors.forename: Robin C
authors.initials: RC
authors.lastname: Spiller
authors.forename: William A
authors.initials: WA
authors.lastname: Stack
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hawkey
mesh_terms: D000230:Adenocarcinoma / Q000473:pathology*; D018988:Bacterial Translocation / Q000502:physiology*; D016123:Campylobacter jejuni / Q000472:pathogenicity*; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000502:physiology*; D003110:Colonic Neoplasms / Q000473:pathology*; D015232:Dinoprostone / Q000378:metabolism; D004847:Epithelial Cells / Q000382:microbiology* / Q000473:pathology*; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008353:Mannitol / Q000378:metabolism; D008954:Models, Biological
chemical_list: D016209:Interleukin-8; D008353:Mannitol; D007770:L-Lactate Dehydrogenase; D015232:Dinoprostone