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Infection and Immunity, May 1999, p. 2575-2584, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Host Cell Death due to Enteropathogenic Escherichia coli Has Features of Apoptosis

John K. Crane,* Swastika Majumdar, and Donald F. Pickhardt III

Center for Microbial Pathogenesis and Division of Infectious Diseases, State University of New York at Buffalo, Buffalo, New York

Received 21 December 1998/Accepted 9 February 1999

Enteropathogenic Escherichia coli (EPEC) is a cause of prolonged watery diarrhea in children in developing countries. The ability of EPEC to kill host cells was investigated in vitro in assays using two human cultured cell lines, HeLa (cervical) and T84 (colonic). EPEC killed epithelial cells as assessed by permeability to the vital dyes trypan blue and propidium iodide. In addition, EPEC triggered changes in the host cell, suggesting apoptosis as the mode of death; such changes included early expression of phosphatidylserine on the host cell surface and internucleosomal cleavage of host cell DNA. Genistein, an inhibitor of tyrosine kinases, and wortmannin, an inhibitor of host phosphatidylinositol 3-kinase, markedly increased EPEC-induced cell death and enhanced the features of apoptosis. EPEC-induced cell death was contact dependent and required adherence of live bacteria to the host cell. A quantitative assay for EPEC-induced cell death was developed by using the propidium iodide uptake method adapted to a fluorescence plate reader. With EPEC, the rate and extent of host cell death were less that what has been reported for Salmonella, Shigella, and Yersinia, three other genera of enteric bacteria known to cause apoptosis. However, rapid apoptosis of the host cell may not favor the pathogenic strategy of EPEC, a mucosa-adhering, noninvasive pathogen.

* Corresponding author. Mailing address: Room 109, Biomedical Research Bldg., 3435 Main St., Buffalo, NY 14214. Phone: (716) 829-2676. Fax: (716) 829-3889. E-mail: jcrane{at}acsu.buffalo.edu.

Infection and Immunity, May 1999, p. 2575-2584, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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