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Infection and Immunity, February 2005, p. 1034-1043, Vol. 73, No. 2
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.2.1034-1043.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Temporal Expression of Enteropathogenic Escherichia coli Virulence Genes in an In Vitro Model of Infection

Laura Q. Leverton1,2 and James B. Kaper1,2*

Department of Microbiology and Immunology,1 Center for Vaccine Development, School of Medicine, University of Maryland, Baltimore, Maryland2

Received 9 April 2004/ Returned for modification 28 July 2004/ Accepted 11 October 2004

The hallmark of enteropathogenic Escherichia coli (EPEC) infection is the ability of EPEC to cause attaching and effacing (A/E) lesions on intestinal epithelium. This event is reproducible in in vitro tissue culture models of infection. We used real-time PCR to measure transcription from several locus of enterocyte effacement (LEE) operons (LEE1 to LEE5) and from bfp during a 5-h infection of HEp-2 cells with EPEC. We found that after the initial formation of A/E lesions, which occurs as early as 5 min postinfection, EPEC continues to increase transcription from LEE3 to LEE5 as well as from bfp. These levels are maximized by 3 h postinfection and remain constant throughout the course of infection. This increase in transcription from LEE3 to LEE5 occurs when LEE1 (ler) transcription is decreasing. EspA, EspB, intimin, Tir, and bundle-forming pilus expression is detectable during the entire 5-h infection. These results indicate that the EPEC genes involved in localized and intimate adherence are continually expressed after the initial stages of A/E lesion formation on host cells.

* Corresponding author. Mailing address: Center for Vaccine Development, School of Medicine, University of Maryland, Baltimore, MD 21201. Phone: (410) 706-2493. Fax: (410) 706-0182. E-mail: jkaper{at}umaryland.edu.

Editor: V. J. DiRita

Infection and Immunity, February 2005, p. 1034-1043, Vol. 73, No. 2
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.2.1034-1043.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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