title: The toxin-coregulated pilus (TCP) of Vibrio cholerae: molecular cloning of genes involved in pilus biosynthesis and evaluation of TCP as a protective antigen in the infant mouse model.
abstract: A serum containing antibodies to non-lipopolysaccharide (non-LPS) protective antigens of Vibrio cholerae has been used, after extensive absorption, to facilitate the cloning of genes involved in the synthesis of toxin-coregulated pili (TCP). A gene bank was constructed from V. cholerae Z17561 DNA using a mobilizable cosmid vector in Escherichia coli, and subsequently transferred by conjugation into V. cholerae O17. This strain does not produce TCP in vitro and lacks non-LPS protective antigens. Eight positive clones were isolated, and of these, four produced TCP as determined by electron microscopic and immunoblotting analyses. TCP-positive O17 clones were 70-fold more virulent than TCP-negative clones or O17 in the infant mouse cholera model. Only the former could remove protective antibodies from the clone-probing serum by absorption. As a corollary, serum containing antibodies to TCP protected mice from challenge with TCP-positive clones, but not with TCP-negative clones or O17. Our data indicate that TCP can function as both a virulence determinant and a protective antigen in the infant mouse model.
authors.affiliation: Department of Microbiology and Immunology, University of Adelaide, South Australia.
authors.forename: D P
authors.initials: DP
authors.lastname: Sharma
authors.forename: U H
authors.initials: UH
authors.lastname: Stroeher
authors.forename: C J
authors.initials: CJ
authors.lastname: Thomas
authors.forename: P A
authors.initials: PA
authors.lastname: Manning
authors.forename: S R
authors.initials: SR
authors.lastname: Attridge
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics*; D002771:Cholera / Q000276:immunology*; D002772:Cholera Toxin; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D010861:Fimbriae, Bacterial / Q000276:immunology*; D051379:Mice; D014734:Vibrio cholerae / Q000235:genetics* / Q000276:immunology; D014774:Virulence / Q000235:genetics
chemical_list: D000942:Antigens, Bacterial; D002772:Cholera Toxin