title: Intimin from enteropathogenic Escherichia coli restores murine virulence to a Citrobacter rodentium eaeA mutant: induction of an immunoglobulin A response to intimin and EspB.
abstract: The formation of attaching and effacing (A/E) lesions is central to the pathogenesis of enteropathogenic Escherichia coli (EPEC)-mediated disease in humans and Citrobacter rodentium (formerly C. freundii biotype 4280)-mediated transmissible colonic hyperplasia in mice. Closely related outer membrane proteins, known as intimins, are required for formation of the A/E lesion by both EPEC (Int(EPEC)) and C. rodentium (Int(CR)). A secreted protein, EspB (formally EaeB), is also necessary for A/E-lesion formation. Here we report that expression of a cloned Int(EPEC), encoded by plasmid pCVD438, restores murine virulence to an intimin-deficient mutant of C. rodentium DBS255. Replacement of Cys937 with Ala abolished the ability of the cloned EPEC intimin to complement the deletion mutation in DBS255. Ultrastructural examination of tissues from wild-type C. rodentium and DBS255(pCVD438)-infected mice revealed multiple A/E lesion on infected cells and loss of contact between enterocytes and basement membrane. Histological investigation showed that although both wild-type C. rodentium and DBS255(pCVD438) colonized the descending colon and induced colonic hyperplasia in orally infected 21-day-old mice, the latter strain adhered to epithelial cells located deeper within crypts. Nonetheless, infection with the wild-type strain was consistently more virulent, as indicated by a higher mortality rate. All the surviving mice, challenged with either wild-type C. rodentium or DBS255(pCVD438), developed a mucosal immunoglobulin A response to intimin and EspB. These results show that C. rodentium infection provides a relevant, simple, and economic model to investigate the role of EPEC proteins in the formation of A/E lesions in vivo and in intestinal disease.
authors.affiliation: Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, United Kingdom. g.frankel@ic.ac.uk
authors.forename: G
authors.initials: G
authors.lastname: Frankel
authors.forename: A D
authors.initials: AD
authors.lastname: Phillips
authors.forename: M
authors.initials: M
authors.lastname: Novakova
authors.forename: H
authors.initials: H
authors.lastname: Field
authors.forename: D C
authors.initials: DC
authors.lastname: Candy
authors.forename: D B
authors.initials: DB
authors.lastname: Schauer
authors.forename: G
authors.initials: G
authors.lastname: Douce
authors.forename: G
authors.initials: G
authors.lastname: Dougan
mesh_terms: D018829:Adhesins, Bacterial; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000633:toxicity*; D002352:Carrier Proteins; D002954:Citrobacter / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D003106:Colon / Q000382:microbiology* / Q000473:pathology; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D006801:Humans; D006965:Hyperplasia; D007070:Immunoglobulin A / Q000096:biosynthesis; D051379:Mice; D009154:Mutation
chemical_list: D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; C087863:EaeB protein, E coli; D029968:Escherichia coli Proteins; D007070:Immunoglobulin A; C073819:eaeA protein, E coli