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Infection and Immunity, April 2003, p. 2262-2265, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2262-2265.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Polymorphisms within EspA Filaments of Enteropathogenic and Enterohemorrhagic Escherichia coli

Bianca C. Neves,1 Robert K. Shaw,2 Gad Frankel,1 and Stuart Knutton2*

Centre for Molecular Microbiology and Infection, Department of Biological Sciences, Imperial College London, London SW7 2AZ,1 Institute of Child Health, University of Birmingham, Birmingham B4 6NH, United Kingdom2

Received 16 September 2002/ Returned for modification 25 November 2002/ Accepted 18 December 2002

Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) possess a filamentous type III secretion system (TTSS) employed to deliver effector proteins into host cells. EspA is a type III secreted protein which forms the filamentous extension to the TTSS and which interacts with host cells during early stages of attaching and effacing (A/E) lesion formation. By immunofluorescence, a polyclonal antibody previously raised to EspA from EPEC strain E2348/69 (O127:H6) stained ~12-nm-diameter EspA filaments produced by this strain but did not stain similar filaments produced by EHEC serotype O157:H7. Similarly, an antibody that we subsequently raised to EHEC strain 85-170 (O157:H7) EspA stained ~12-nm-diameter EspA filaments produced by strain 85-170 but did not stain E2348/69 EspA filaments. Given such heterogeneity between EPEC and EHEC EspA filaments, we examined polymorphisms of functional EspA filaments among different EPEC and EHEC serotypes. With use of the EPEC EspA antiserum, EspA filaments were observed only with EPEC serotypes O127:H6 and O55:H6, serotypes which encode an identical EspA protein. When stained with the EHEC EspA antiserum, EspA filaments were detected only on EHEC strains belonging to serotype O157:H7; the EHEC antiserum did, however, stain EspA filaments produced by the closely related EPEC serotype O55:H7 but not filaments of any other EPEC serotype tested. Such polymorphisms among functional EspA filaments of EPEC and EHEC would be expected to have important implications for the development of broad-range EspA-based vaccines.

* Corresponding author. Mailing address: Institute of Child Health, Whittall St., Birmingham B4 6NH, United Kingdom. Phone: 44 121 333 8746. Fax: 44 121 333 8701. E-mail: s.knutton{at}bham.ac.uk.

Editor: A. D. O'Brien

Infection and Immunity, April 2003, p. 2262-2265, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2262-2265.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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