title: Detection of intimins alpha, beta, gamma, and delta, four intimin derivatives expressed by attaching and effacing microbial pathogens.
abstract: Intimins are outer membrane proteins expressed by enteric bacterial pathogens capable of inducing intestinal attachment-and-effacement lesions. A eukaryotic cell-binding domain is located within a 280-amino-acid (Int280) carboxy terminus of intimin polypeptides. Polyclonal antiserum was raised against Int280 from enteropathogenic Escherichia coli (EPEC) serotypes O127:H6 and O114:H2 (anti-Int280-H6 and anti-Int280-H2, respectively), and Western blot analysis was used to explore the immunological relationship between the intimin polypeptides expressed by different clinical EPEC and enterohemorrhagic E. coli (EHEC) isolates, a rabbit diarrheagenic E. coli strain (RDEC-1), and Citrobacter rodentium. Anti-Int280-H6 serum reacted strongly with some EPEC serotypes, whereas anti-Int280-H2 serum reacted strongly with strains belonging to different EPEC and EHEC serotypes, RDEC-1, and C. rodentium. These observations were confirmed by using purified Int280 in an enzyme-linked immunosorbent assay and by immunogold and immunofluorescence labelling of whole bacterial cells. Some bacterial strains were recognized poorly by either antiserum (e.g., EPEC O86:H34 and EHEC O157:H7). By using PCR primers designed on the basis of the intimin-encoding eae gene sequences of serotype O127:H6, O114:H2, and O86:H34 EPEC and serotype O157:H7 EHEC, we could distinguish between different eae gene derivatives. Accordingly, the different intimin types were designated alpha, beta, delta, and gamma, respectively.
authors.affiliation: Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, United Kingdom.
authors.forename: J
authors.initials: J
authors.lastname: Adu-Bobie
authors.forename: G
authors.initials: G
authors.lastname: Frankel
authors.forename: C
authors.initials: C
authors.lastname: Bain
authors.forename: A G
authors.initials: AG
authors.lastname: Goncalves
authors.forename: L R
authors.initials: LR
authors.lastname: Trabulsi
authors.forename: G
authors.initials: G
authors.lastname: Douce
authors.forename: S
authors.initials: S
authors.lastname: Knutton
authors.forename: G
authors.initials: G
authors.lastname: Dougan
mesh_terms: D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D000818:Animals; D000940:Antigenic Variation; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002352:Carrier Proteins; D002954:Citrobacter / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D029968:Escherichia coli Proteins; D005455:Fluorescent Antibody Technique; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry; D016133:Polymerase Chain Reaction; D011817:Rabbits; D016415:Sequence Alignment; D012703:Serotyping; D014407:Tumor Cells, Cultured
chemical_list: D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D007106:Immune Sera; C073819:eaeA protein, E coli