HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by TAKAHASHI, A. Articles by HONDA, T. Search for Related Content PubMed PubMed Citation Articles by TAKAHASHI, A. Articles by HONDA, T. Agricola Articles by TAKAHASHI, A. Articles by HONDA, T.

Mechanisms of chloride secretion induced by thermostable direct haemolysin of Vibrio parahaemolyticus in human colonic tissue and a human intestinal epithelial cell line

Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, 3-1, Yamadaoka, Suita, Osaka 565-0871 and *Department of Urology, School of Medicine, University of Kobe, 7-5-1 Kusunoki, Tyuoku, Kobe, Hyogo 650-0017, Japan

Corresponding author: Dr A. Takahashi (e-mail: akiratak{at}nutr.med.tokushima-u.ac.jp).

Received 9 Aug. 1999; revised version accepted 31 Jan. 2000.

Abstract

Thermostable direct haemolysin (TDH) produced by Vibrio parahaemolyticus is thought to play an important role in the severe diarrhoea caused by this organism. This study investigated the enterotoxicity of TDH for human intestinal cells. Addition of TDH to the mucosal side of human colonic tissue in Ussing chambers caused increased short circuit currents (Isc), a process that was inhibited by 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), an inhibitor of Ca²⁺-activated chloride (Cl^-) channels. With human colonic epithelial (Caco-2) cells, high Isc and intracellular Ca²⁺ concentrations ([Ca²⁺]_in) were detected after the addition of TDH to the apical side of the cell monolayer. The Isc decreased with the addition of DIDS, but not with glybenclamide, 5-nitro-2-(3-phenylpropylamino) benzoic acid, or gadolinium chloride. No Isc increase with TDH was observed when the Cl^- in the medium was replaced by gluconate or when Ca²⁺ was depleted. Similarly, TDH did not raise [Ca²⁺]_in after depletion of extracellular Ca²⁺. R7, a mutant form of TDH, reduced the effects of TDH on Isc and [Ca²⁺]_in, as did protein kinase C (PKC) inhibitors. Thus, TDH increases Cl^- secretion in human colonic epithelial cells, apparently through mechanisms involving cell binding and Ca²⁺ influx, followed by elevation of [Ca²⁺]_in associated with PKC phosphorylation.

This article has been cited by other articles:

				R. De La Fuente, W. Namkung, A. Mills, and A. S. Verkman Small-Molecule Screen Identifies Inhibitors of a Human Intestinal Calcium-Activated Chloride Channel Mol. Pharmacol., March 1, 2008; 73(3): 758 - 768. [Abstract] [Full Text] [PDF]

				A. Takahashi, C. Yamamoto, T. Kodama, K. Yamashita, N. Harada, M. Nakano, T. Honda, and Y. Nakaya Pore Formation of Thermostable Direct Hemolysin Secreted from Vibrio parahaemolyticus in Lipid Bilayers International Journal of Toxicology, September 1, 2006; 25(5): 409 - 418. [Abstract] [Full Text] [PDF]

				T. Lynch, S. Livingstone, E. Buenaventura, E. Lutter, J. Fedwick, A. G. Buret, D. Graham, and R. DeVinney Vibrio parahaemolyticus Disruption of Epithelial Cell Tight Junctions Occurs Independently of Toxin Production Infect. Immun., March 1, 2005; 73(3): 1275 - 1283. [Abstract] [Full Text] [PDF]

				J Berkes, V K Viswanathan, S D Savkovic, and G Hecht Intestinal epithelial responses to enteric pathogens: effects on the tight junction barrier, ion transport, and inflammation Gut, March 1, 2003; 52(3): 439 - 451. [Abstract] [Full Text] [PDF]

				A. TAKAHASHI, T. IIDA, R. NAIM, Y. NAYKAYA, and T. HONDA Chloride secretion induced by thermostable direct haemolysin of Vibrio parahaemolyticus depends on colonic cell maturation J. Med. Microbiol., October 1, 2001; 50(10): 870 - 878. [Abstract] [Full Text] [PDF]