Study of Helicobacter pullorum proinflammatory properties on human epithelial cells in vitro.
OBJECTIVE
Helicobacter pullorum is an enterohepatic Helicobacter species of avian origin detected in patients with acute diarrhoea and inflammatory bowel disease. The aim of the present study was to determine whether H pullorum exerts a direct effect on human intestinal epithelial cells in vitro and to characterise the bacterial mechanisms and the signalling pathways involved.


METHODS
The proinflammatory properties of H pullorum from human and avian origins were measured on human gastric (AGS) and intestinal (CaCo-2 and HT-29) epithelial cell lines after co-culture with different H pullorum strains, and the extent of nuclear factor-kappaB (NF-kappaB) involvement was determined.


RESULTS
All of the H pullorum strains tested stimulated interleukin 8 (IL8) secretion by the three cell lines. Similar results were obtained with heat-killed H pullorum. Incubation of cells with filtered H pullorum culture supernatants did not stimulate IL8 secretion. The same observation was made when bacterial adherence was inhibited by Transwell inserts. H pullorum induced NF-kappaB activation and rapid nuclear translocation as demonstrated by immunofluorescent staining and cellular fractionation. NF-kappaB involvement was confirmed by using the specific inhibitor SN50 and small interfering RNA (siRNA) which abolished H pullorum-induced IL8 production.


CONCLUSIONS
H pullorum strains stimulate IL8 secretion by human gastric and intestinal epithelial cell lines. This effect requires bacterial adherence and probably lipopolysaccharides, and is mediated by NF-kappaB signalling. The present study strengthens the argument that H pullorum is a potent human pathogen and highlights its putative role in acute and chronic digestive diseases such as inflammatory bowel disease.
Varon
C
C
INSERM U853, Bordeaux, France.
Duriez
A
A
Lehours
P
P
Ménard
A
A
Layé
S
S
Zerbib
F
F
Mégraud
F
F
Laharie
D
D
D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology; D015153:Blotting, Western; D018938:Caco-2 Cells; D002467:Cell Nucleus / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology*; D019073:HT29 Cells; D016998:Helicobacter / Q000472:pathogenicity*; D016480:Helicobacter pylori / Q000472:pathogenicity; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D011200:Poultry / Q000382:microbiology; D015398:Signal Transduction
D016209:Interleukin-8; D016328:NF-kappa B