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Chloride secretion induced by thermostable direct haemolysin of Vibrio parahaemolyticus depends on colonic cell maturation

Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, 3-1, Yamadaoka, Suita, Osaka 565-0871 and *Department of Nutrition, School of Medicine, Tokushima University, Kuramoto-cho, 3-15-18, Tokushima 770-8503, Japan

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

Received 4 Sept. 2000; revised version received 15 March 2001; accepted 9 April 2001.

Abstract

Vibrio parahaemolyticus produces a thermostable direct haemolysin (TDH) that has been implicated in the pathogenesis of diarrhoeal disease caused by this organism. In previous work, TDH induced Cl^- secretion in human colonic epithelial cells that was dependent on the intracellular Ca²⁺ concentration, [Ca²⁺]_in. This study investigated whether Cl^- secretion induced by TDH is influenced by the stage of maturation of intestinal epithelial cells. Two different human colonic cell lines, villus cell-like Caco-2 cells and crypt cell-like T84 cells, cultured by different methods to obtain differentiated samples, were used. When these cells were exposed to butyrate, a transcriptional regulator of differentiation genes, or co-cultured with 18Co cells, a human colonic fibroblast cell line, they showed increased trans-epithelial resistance and villus cell marker enzyme activity. In Caco-2 cells, exposure to butyrate or co-culturing with 18Co cells resulted in increased TDH binding, higher short-circuit currents (Isc) and greater [Ca²⁺]_in. These results suggest that sensitivity to TDH is affected by the stage of cellular differentiation of cultured intestinal epithelial cells.

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