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Infection and Immunity, December 1998, p. 5643-5649, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification of Immunodominant Regions within the C-Terminal Cell Binding Domain of Intimin alpha  and Intimin beta  from Enteropathogenic Escherichia coli

Jeannette Adu-Bobie,1 Luiz R. Trabulsi,2 Magda M. S. Carneiro-Sampaio,3 Gordon Dougan,1 and Gad Frankel1,*

Department of Biochemistry, Imperial College of Science, Technology and Medicine, London SW7 2AZ, United Kingdom,1 and Departamentos de Microbiologia2 e Immunologia,3 Instituto de Ciencias Biomedicas, Universidade de São Paulo, São Paulo Cep 05508-900, Brazil

Received 15 June 1998/Returned for modification 25 August 1998/Accepted 18 September 1998

Enteropathogenic Escherichia coli (EPEC) strains are a common cause of infantile diarrhea in developing countries. EPEC strains induce a characteristic attaching and effacing (A/E) lesion on epithelial cells. A/E lesion formation requires intimin, an outer membrane adhesin protein. The cell-binding activity of intimin is localized at the C-terminal 280 amino acids of the polypeptide (Int280). So far, four distinct Int280 types (alpha , beta , gamma , and delta ) have been identified. The aim of this study was to identify immunodominant regions within the Int280alpha and Int280beta domains. Recombinant DNA was used to construct and express overlapping polypeptides spanning these domains. Rabbit anti-Int280 antisera and human colostral immunoglobulin A were reacted with these polypeptides in Western blots and enzyme-linked immunosorbent assays. The results obtained with the rabbit antisera showed the presence of two separate immunodominant regions which are common to both Int280alpha and Int280beta . The first localized within the N-terminal region of Int280, and the second localized between amino acids 80 and 130. The results with the human colostra revealed one reactivity pattern against the Int280alpha fragments but two different reactivity patterns against the Int280beta domain.

* Corresponding author. Mailing address: Department of Biochemistry, Imperial College of Science, Technology and Medicine, London SW7 2AZ, United Kingdom. Phone: 44-171-594-5253. Fax: 44-171-594-5255. E-mail: g.frankel{at}ic.ac.uk.

Infection and Immunity, December 1998, p. 5643-5649, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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