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Infection and Immunity, March 2005, p. 1275-1283, Vol. 73, No. 3
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.3.1275-1283.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Vibrio parahaemolyticus Disruption of Epithelial Cell Tight Junctions Occurs Independently of Toxin Production

Tarah Lynch,1 Scott Livingstone,1 Enrico Buenaventura,2 Erika Lutter,1 Jason Fedwick,3 Andre G. Buret,3 David Graham,2 and Rebekah DeVinney1*

Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre,1 Department of Biological Sciences, University of Calgary, Calgary,3 Canadian Food Inspection Agency, Burnaby, Canada2

Received 22 June 2004/ Returned for modification 26 July 2004/ Accepted 28 October 2004

Vibrio parahaemolyticus is a leading cause of seafood-borne gastroenteritis worldwide. Virulence is commonly associated with the production of two toxins, thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH). Although the majority of clinical isolates produce TDH and/or TRH, clinical samples lacking toxin genes have been identified. In the present study, we investigated the effects of V. parahaemolyticus on transepithelial resistance (TER) and paracellular permeability in Caco-2 cultured epithelial cells. We found that V. parahaemolyticus profoundly disrupts epithelial barrier function in Caco-2 cells and that this disruption occurs independently of toxin production. Clinical isolates with different toxin genotypes all led to a significant decrease in TER, which was accompanied by an increased flux of fluorescent dextran across the Caco-2 monolayer, and profound disruption of actin and the tight junction-associated proteins zonula occludin protein 1 and occludin. Purified TDH, even at concentrations eightfold higher than those produced by the bacteria, had no effect on either TER or paracellular permeability. We used lactate dehydrogenase release as a measure of cytotoxicity and found that this parameter did not correlate with the ability to disrupt tight junctions. As the effect on barrier function occurs independently of toxin production, we used PCR to determine the toxin genotypes of V. parahaemolyticus isolates obtained from both clinical and environmental sources, and we found that 5.6% of the clinical isolates were toxin negative. These data strongly indicate that the effect on tight junctions is not due to TDH and suggest that there are other virulence factors.


* Corresponding author. Mailing address: Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, 3330 Hospital Dr. NW, Calgary AB, T2N 4N1 Canada. Phone: (403) 220-4095. Fax: (403) 270-2772. E-mail: rdevinne{at}ucalgary.ca.

Editor: F. C. Fang


Infection and Immunity, March 2005, p. 1275-1283, Vol. 73, No. 3
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.3.1275-1283.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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