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Infect Immun, April 1998, p. 1680-1687, Vol. 66, No. 4
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Signal Transduction Pathways Involved in Enterohemorrhagic Escherichia coli-Induced Alterations in T84 Epithelial Permeability

Dana J. Philpott,1,2,3 Derek M. McKay, Walter Mak,4 Mary H. Perdue, and Philip M. Sherman1,2,4,*

Division of Gastroenterology and Nutrition, Research Institute, The Hospital for Sick Children,4 and Departments of Pediatrics1 and Molecular and Medical Genetics,2 University of Toronto, Toronto, and Intestinal Disease Research Programme, McMaster University, Hamilton,3 Ontario, Canada

Received 24 September 1997/Returned for modification 4 November 1997/Accepted 14 January 1998

Enterohemorrhagic Escherichia coli (EHEC) infection is associated with watery diarrhea and can lead to complications, including hemorrhagic colitis and the hemolytic-uremic syndrome. The mechanisms by which these organisms produce diarrheal disease remain to be elucidated. Changes in T84 epithelial cell electrophysiology were examined following EHEC infection. T84 cell monolayers infected with EHEC O157:H7 displayed a time-dependent decrease in transepithelial resistance. Increases in the transepithelial flux of both [3H]mannitol and 51Cr-EDTA accompanied the EHEC-induced decreases in T84 resistance. Altered barrier function induced by EHEC occurred at the level of the tight junction since immunofluorescent staining of the tight-junction-associated protein ZO-1 was disrupted when examined by confocal microscopy. Decreased resistance induced by EHEC involved a protein kinase C (PKC)-dependent pathway as the highly specific PKC inhibitor, CGP41251, abrogated the EHEC-induced drop in resistance. PKC activity was also increased in T84 cells infected with EHEC. Calmodulin and myosin light chain kinase played a role in EHEC-induced resistance changes as inhibition of these effector molecules partially reversed the effects of EHEC on barrier function. These studies demonstrate that intracellular signal transduction pathways activated following EHEC infection link the increases in T84 epithelial permeability induced by this pathogen.

* Corresponding author. Mailing address: Division of Gastroenterology, Room 8411, The Hospital for Sick Children, 555 University Ave., Toronto, Ontario, Canada M5G 1X8. Phone: (416) 813-6185. Fax: (416) 813-6531. E-mail: sherman{at}sickkids.on.ca.




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