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Infection and Immunity, December 2001, p. 7356-7364, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7356-7364.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Induction of Epithelial Cell Death Including Apoptosis by Enteropathogenic Escherichia coli Expressing Bundle-Forming Pili

Maan Abul-Milh,1,2 Ying Wu,1 Bedy Lau,1 Clifford A. Lingwood,2,3 and Debora Barnett Foster1,2,*

Department of Chemistry, Biology and Chemical Engineering, Ryerson University, Toronto, Ontario M5B 2K3,1 Division of Immunity, Infection, Injury and Repair Research Institute, Hospital for Sick Children, Toronto, Ontario M5G 1X8,2 and Departments of Biochemistry and Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5G 1L5,3 Canada

Received 19 March 2001/Returned for modification 14 May 2001/Accepted 17 September 2001

Infection with enteropathogenic Escherichia coli (EPEC) is a major cause of severe infantile diarrhea, particularly in parts of the developing world. The bundle-forming pilus (BFP) of EPEC is an established virulence factor encoded on the EPEC adherence factor plasmid (EAF) and has been implicated in both localized adherence to host cells and bacterial autoaggregation. We investigated the role of BFP in the ability of EPEC binding to kill host epithelial cells. BFP-expressing strains killed all three cell lines tested, comprising HEp-2 (laryngeal), HeLa (cervical), and Caco-2 (colonic) cells. Analysis of phosphatidylserine expression, internucleosomal cleavage of host cell DNA, and morphological changes detected by electron microscopy indicated evidence of apoptosis. The extent of cell death was significantly greater for BFP-expressing strains, including E2348/69, a wild-type clinical isolate, as well as for a laboratory strain, HB101, transformed with a bfp-carrying plasmid. Strains which did not express BFP induced significantly less cell death, including a bfpA disruptional mutant of E2348/69, EAF plasmid-cured E2348/69, HB101, and HB101 complemented with the locus of enterocyte effacement pathogenicity island. These results indicate a direct correlation between BFP expression and induction of cell death, including apoptosis, an event which may involve the targeting of host cell membrane phosphatidylethanolamine.

* Corresponding author. Mailing address: Department of Chemistry, Biology and Chemical Engineering, Ryerson University, 350 Victoria St., Toronto, Ontario M5B 2K3, Canada. Phone: (416) 979-5000, ext. 6345. Fax: (416) 979-5044. E-mail: dfoster{at}acs.ryerson.ca.

Infection and Immunity, December 2001, p. 7356-7364, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7356-7364.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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