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Infection and Immunity, October 1999, p. 5338-5344, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Involvement of the Enteroaggregative Escherichia coli Plasmid-Encoded Toxin in Causing Human Intestinal Damage

Ian R. Henderson,1 Susan Hicks,2 Fernando Navarro-Garcia,1,dagger Waldir P. Elias,1,Dagger Alan D. Philips,2 and James P. Nataro1,*

Center for Vaccine Development, Department of Pediatrics, University of Maryland School of Medicine, Baltimore, Maryland 21201,1 and University Department of Pediatric Gastroenterology, Royal Free Hospital, London, England2

Received 20 May 1999/Returned for modification 21 June 1999/Accepted 26 July 1999

Enteroaggregative Escherichia coli (EAEC) strains have been shown to adhere to human intestinal tissue in an in vitro organ culture (IVOC) model, and certain strains manifest mucosal toxicity. We have recently described the EAEC plasmid-encoded toxin (Pet), a member of a specific serine protease subclass of the autotransporter proteins. When injected into rat ileal loops, Pet both elicited fluid accumulation and had cytotoxic effects on the mucosa. Furthermore, the Pet protein caused rises in short circuit current from rat jejunal tissue mounted in a Ussing chamber and rounding of intestinal epithelial cells in culture. We therefore hypothesized that the mucosal pathology induced by EAEC strains in the IVOC model was related to expression of the Pet protein. Here, we have examined the effects of EAEC strain 042 and its isogenic pet mutant in the IVOC model. 042-infected colonic explants exhibited dilation of crypt openings, increased cell rounding, development of prominent intercrypt crevices, and absence of apical mucus plugs. Colonic tissue incubated with the pet mutant exhibited significantly fewer mucosal abnormalities both subjectively and as quantitated morphometrically by measurement of crypt aperture diameter. Mucosal effects were restored upon complementation of the pet mutation in trans. Interestingly, we found that the ability of 042 to damage T84 cells was not dependent upon Pet. The data suggest that the Pet toxin is active on the human intestinal mucosa but that EAEC may have other mechanisms of eliciting mucosal damage.

* Corresponding author. Mailing address: Center for Vaccine Development, Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD 21201. Phone: (410) 706-7376. Fax: (410) 706-6205. E-mail: jnataro{at}medicine.umaryland.edu.

dagger Present address: Department of Public Health, Faculty of Medicine, UNAM, 04510 Mexico City, Mexico.

Dagger Present address: Laboratório Especial de Microbiologia, Instituto Butantan, São Paulo, Brazil.

Infection and Immunity, October 1999, p. 5338-5344, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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