title: Cloning and immunologic characterization of a truncated Bordetella bronchiseptica filamentous hemagglutinin fusion protein.
abstract: Filamentous hemagglutinin (FHA) is an outer-membrane associated adhesin conserved within the genus Bordetella. FHA provides protection against B. pertussis infections in humans and is a component of acellular whooping cough vaccines. Furthermore, FHA serves as a protective antigen in several animal models of infection with B. bronchiseptica and may serve as a protective antigen of canine bordetellosis. In this study, polyclonal anti-B. pertussis FHA antiserum was used to identify an immunoreactive clone from the genomic DNA library of a canine B. bronchiseptica field isolate. The nucleotide and predicted amino acid sequences of the immunoreactive clone were compared to fhaB and FhaB from B. pertussis revealing 94% identity at the nucleic acid level, and 86% identity at the protein level. A truncated fusion protein (FHAt) was prepared which included a conserved domain homologous to the immunodominant region in the FHA of B. pertussis [Leininger E, Bowen S, Renauld-Mongen G, Rouse JH, Menozzi FD, Locht C, Heron I, Brennan MJ. Immunodominant domain present on the Bordetella pertussis vaccine component filamentous hemagglutinin. J. Infect. Dis. 1997;175:1423-1431; Wilson DR, Siebers A, Finlay BB. Antigenic analysis of Bordetella pertussis filamentous hemagglutinin with phage display libraries and rabbit anti-filamentous hemagglutinin polyclonal antibodies. Infect. Immun. 1998;66:4884-4894]. FHAt was shown to be safe and antigenic in rabbits. FHAt induced the formation of antibodies that inhibit the hemagglutination associated with full length B. pertussis FHA, and inhibit adherence of B. bronchisepitca to canine fibroblasts by as much as 65%. This information may have implications for the development of safe and efficacious subunit vaccines for the prevention of canine bordetellosis and may contribute to future acellular whooping cough vaccines.
authors.affiliation: Department of Diagnostic Medicine/Pathobiology, Kansas State University College of Veterinary Medicine, Manhattan, KS 66506-5606, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Keil
authors.forename: E H
authors.initials: EH
authors.lastname: Burns
authors.forename: W R
authors.initials: WR
authors.lastname: Kisker
authors.forename: D
authors.initials: D
authors.lastname: Bemis
authors.forename: B
authors.initials: B
authors.lastname: Fenwick
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics* / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology*; D001422:Bacterial Adhesion; D001483:Base Sequence; D016950:Bordetella bronchiseptica / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D006385:Hemagglutination Inhibition Tests; D006388:Hemagglutinins / Q000235:genetics* / Q000276:immunology*; D016056:Immunodominant Epitopes; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D010566:Virulence Factors, Bordetella
chemical_list: D018829:Adhesins, Bacterial; D000942:Antigens, Bacterial; D006388:Hemagglutinins; D016056:Immunodominant Epitopes; D011993:Recombinant Fusion Proteins; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis