Detection of intimins alpha, beta, gamma, and delta, four intimin derivatives expressed by attaching and effacing microbial pathogens.
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.
Adu-Bobie
J
J
Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, United Kingdom.
Frankel
G
G
Bain
C
C
Goncalves
A G
AG
Trabulsi
L R
LR
Douce
G
G
Knutton
S
S
Dougan
G
G
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
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D007106:Immune Sera; C073819:eaeA protein, E coli