Differential modulation of NF-κB-mediated pro-inflammatory response in human intestinal epithelial cells by cheY homologues of Vibrio cholerae

Vibrio cholerae, the etiological agent of cholera, colonizes the small intestine, produces an enterotoxin and causes acute inflammatory response at intestinal epithelial surface. Chemotaxis and motility greatly influence the infectivity of V. cholerae although the role of chemotaxis genes in V. cholerae pathogenesis is less well understood. Four cheY genes are present in three clusters in the complete genome sequence of V. cholerae. A less motile and less adherent mutant was generated by inactivation of cheY-3 (O395Y3N) or cheY-4 (O395Y4N) whereas alterations in motility or adherence were not observed for cheY-1 (O395Y1N) or cheY-2 (O395Y2N) insertional mutants. In contrast to O395Y1N and O395Y2N, O395Y3N and O395Y4N showed reduced cholera toxin production compared to wild-type in vitro. Infection of the human intestinal epithelial cell line Int407 with O395Y3N and O395Y4N caused reduced secretion of interleukin (IL)-1a, IL-6, tumor necrosis factor (TNF-a) and monocyte chemotactic protein-1 (MCP-1) compared to wild-type and was associated with delayed activation of nuclear factor kappa B (NF-κB) p65 and its co-activator cAMP response element binding protein (CREB). Further, the absence of nuclear translocation of NF-κB p50 subunit upon infection with O395Y3N or O395Y4N and its reversal upon complementation indicates the involvement of cheY-3 and cheY-4 in V. cholerae-induced pro-inflammatory response in the INT407 cell line.