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Infection and Immunity, September 2000, p. 5090-5095, Vol. 68, No. 9
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Human Response to Escherichia coli O157:H7 Infection: Antibodies to Secreted Virulence Factors

Yuling Li,1 Elizabeth Frey,1 Andrew M. R. Mackenzie,2,dagger and B. Brett Finlay1,*

Biotechnology Laboratory, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3,1 and Division of Microbiology, Ottawa Hospital Civic Campus, Ottawa, Ontario, Canada KIY 4E92

Received 10 April 2000/Returned for modification 9 May 2000/Accepted 2 June 2000

Vaccination has been proposed for the prevention of disease due to enterohemorrhagic Escherichia coli (EHEC), but the immune response following human infection, including the choice of potential antigens, has not been well characterized. To study this, sera were obtained from five pediatric patients with acute diarrhea caused by E. coli O157:H7 0, 8, and 60 days after hospitalization. These sera were used to examine the immune response to four different EHEC virulence factors: Tir (translocated intimin receptor, which is inserted into the host cell membrane), intimin (bacterial outer membrane protein which binds to Tir), EspA (secreted protein which forms filamentous structures on EHEC surface), and EspB (inserted into the host membrane and cytoplasm). The response to O157:H7 lipopolysaccharide was also examined. Sera were assayed against purified recombinant proteins using immunoblot analysis and by enzyme-linked immunosorbent assay to determine the sera's titers to each of the antigens in all patients. We found that there was little reaction to EspA, EspB, and intimin in the acute-phase sera, although there was some reactivity to Tir. By day 8, titers of antibody to all four virulence factors were present in all patients, with a very strong response against Tir (up to a titer of 1:256,000), especially in hemolytic-uremic syndrome patients, and lesser strong responses to the other three antigens. The titer to the antigens 60 days after hospitalization was decreased but was still highest for Tir. These results suggest that there is a strong immune response to Tir, and to a lesser extent to the other three virulence factors, following EHEC disease, indicating that these bacterial molecules are potential vaccine candidates for preventing EHEC disease. They also suggest that bacterial virulence factors that are inserted into host cells during infection by type III secretion systems (Tir or EspB) are still recognized by the host immune response.

* Corresponding author. Mailing address: Biotechnology Laboratory, University of British Columbia, Room 237-Wesbrook Building, 6174 University Blvd., Vancouver, British Columbia, Canada V6T-1Z3. Phone: (604) 822-2210. Fax: (604) 822-9830. E-mail: bfinlay{at}interchange.ubc.ca.

dagger Present address: Child and Youth Clinical Trial Network, Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario, Canada KIH 8L1.

Infection and Immunity, September 2000, p. 5090-5095, Vol. 68, No. 9
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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