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IL-1beta expression in Int407 is induced by flagellin of Vibrio cholerae through TLR5 mediated pathway
Arunava Bandyopadhaya1, Madhubanti Sarkar, Keya Chaudhuri
1Molecular and Human Genetics Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Kolkata 700 032, India.
Abstract:
Vibrio cholerae, a noninvasive enteric bacterium, causing inflammatory diarrheal disease cholera, is associated with the secretion of proinflamammatory cytokines including IL-1beta in cultured epithelial cells. Incubation of Int407 with live V. cholerae resulted in increased IL-1beta mRNA expression as early as 2h of infection, reached a peak at approximately 3.5h and decreased thereafter. The identity of the effector molecule(s) is largely unknown. The bacterial culture supernatant showed IL-1beta stimulating activity. An engineered aflagellate V. cholerae flaA mutant (O395FLAN) resulted in highly reduced level of IL-1beta expression in Int407. The crude flagellar protein of V. cholerae as well as recombinant FlaA induced IL-1beta expression in Int407. Infection of Toll-like receptor 5 (TLR5) transfected HeLa cells with O395FLAN showed reduced expression of IL-1beta compared to wild-type. Unlike wild-type V. cholerae, O395FLAN did not activate the NF-kappaB while the recombinant flagellin could activate NF-kappaB. Finally, the mitogen activated protein kinases (ERK1 and 2, p38) were phosphorylated in wild-type and recombinant flagellin treated Int407 cells and inhibition of the p38 and ERK pathways significantly decreased the IL-1beta response induced by wild-type V. cholerae as well as recombinant flagellin. Our data clearly indicate that flagellin of V. cholerae could induce IL-1beta expression by recognizing TLR5 that activate NF-kappaB and MAP kinase in Int407.
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