Vibrio parahaemolyticus disruption of epithelial cell tight junctions occurs independently of toxin production.
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.
Lynch
Tarah
T
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, 3330 Hospital Dr. NW, Calgary AB, T2N 4N1 Canada.
Livingstone
Scott
S
Buenaventura
Enrico
E
Lutter
Erika
E
Fedwick
Jason
J
Buret
Andre G
AG
Graham
David
D
DeVinney
Rebekah
R
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability; D004847:Epithelial Cells / Q000382:microbiology / Q000473:pathology*; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010125:Ostreidae / Q000382:microbiology; D012623:Seawater / Q000382:microbiology; D012758:Shellfish / Q000382:microbiology; D019108:Tight Junctions / Q000382:microbiology / Q000473:pathology*; D014735:Vibrio Infections / Q000382:microbiology; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D014774:Virulence
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; C088609:thermostable direct hemolysin-related hemolysin protein, Vibrio parahaemolyticus; C068489:thermostable direct hemolysin
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