Effect of Helicobacter pylori on polymorphonuclear leukocyte migration across polarized T84 epithelial cell monolayers: role of vacuolating toxin VacA and cag pathogenicity island.
Helicobacter pylori infection can induce polymorphonuclear leukocyte (PMNL) infiltration of the gastric mucosa, which characterizes acute chronic gastritis. The mechanisms underlying this process are poorly documented. The lack of an in vitro model has considerably impaired the study of transepithelial migration of PMNL induced by H. pylori. In the present work, we used confluent polarized monolayers of the human intestinal cell line T84 grown on permeable filters to analyze the epithelial PMNL response induced by broth culture filtrates (BCFs) and bacterial suspensions from different strains of H. pylori. We have evaluated the role of the vacuolating cytotoxin VacA and of the cag pathogenicity island (PAI) of H. pylori in PMNL migration via their effects on T84 epithelial cells. We noted no difference in the rates of PMNL transepithelial migration after epithelial preincubation with bacterial suspensions or with BCFs of VacA-negative or VacA-positive H. pylori strains. In contrast, PMNL transepithelial migration was induced after incubation of the T84 cells with cag PAI-positive and cagE-positive H. pylori strains. Finally, PMNL migration was correlated with a basolateral secretion of interleukin-8 by T84 cells, thus creating a subepithelial chemotactic gradient for PMNL. These data provide evidence that the vacuolating cytotoxin VacA is not involved in PMNL transepithelial migration and that the cag PAI, with a pivotal role for the cagE gene, provokes a transcellular signal across T84 monolayers, inducing a subepithelial PMNL response.
Hofman
V
V
Laboratoire d'Anatomie-Pathologique, IFR 50, Faculté de Médecine, 06107 Nice Cedex 01, France.
Ricci
V
V
Galmiche
A
A
Brest
P
P
Auberger
P
P
Rossi
B
B
Boquet
P
P
Hofman
P
P
D000942:Antigens, Bacterial; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000502:physiology*; D002465:Cell Movement; D016764:Cell Polarity; D016480:Helicobacter pylori / Q000472:pathogenicity*; D006801:Humans; D016209:Interleukin-8 / Q000502:physiology; D009504:Neutrophils / Q000502:physiology*; D014407:Tumor Cells, Cultured
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D016209:Interleukin-8; C074689:VacA protein, Helicobacter pylori; C080866:cagA protein, Helicobacter pylori