title: Identification and characterization of a protective immunodominant B cell epitope of pertactin (P.69) from Bordetella pertussis.
abstract: Epitopes defined by monoclonal antibodies (mAb) specific for the Bordetella pertussis outer membrane protein P.69 (pertactin) were mapped using a series of amino- and carboxy-terminal deletion mutants expressed in Escherichia coli. mAb were found to bind predominantly to a region of pertactin spanning a (Pro-Gln-Pro)5 repeat motif and one mAb was found to bind to another region spanning a (Gly-Gly-Xaa-Xaa-Pro)5 repeat motif. To localize further the mAb-binding sites, a panel of synthetic peptides, a series of 94 overlapping hexameric peptides, and a P.69 30-amino acid fusion to a hepatitis B core protein (HBcAg-69), were synthesized. This combined approach has identified the binding site for the mAb BBO5: Pro-Gly-Pro-Gln-Pro-Pro; mAb BBO7, E4A8 and E4D7: Ala-Pro-Gln-Pro-Pro-Ala-Gly-Arg; and mAb BPE3: Thr-Leu-Trp-Tyr-Ala-Glu-Ser-Asn-Ala-Leu-Ser-Lys-Arg. We have used a non-lethal murine respiratory model of B. pertussis infection to investigate the ability of a peptide containing the epitope of the mAb BBO5 to elicit protective immunity. Immunization of mice with the HBcAg-69 protein prevented growth of B. pertussis in the lungs compared to mice receiving HBcAg alone, and protection correlated with high titers of anti-P.69 antibodies.
authors.affiliation: Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, GB.
authors.forename: I G
authors.initials: IG
authors.lastname: Charles
authors.forename: J L
authors.initials: JL
authors.lastname: Li
authors.forename: M
authors.initials: M
authors.lastname: Roberts
authors.forename: K
authors.initials: K
authors.lastname: Beesley
authors.forename: M
authors.initials: M
authors.lastname: Romanos
authors.forename: D J
authors.initials: DJ
authors.lastname: Pickard
authors.forename: M
authors.initials: M
authors.lastname: Francis
authors.forename: D
authors.initials: D
authors.lastname: Campbell
authors.forename: G
authors.initials: G
authors.lastname: Dougan
authors.forename: M J
authors.initials: MJ
authors.lastname: Brennan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology*; D001886:Bordetella pertussis / Q000276:immunology*; D000939:Epitopes / Q000032:analysis*; D006512:Hepatitis B Core Antigens / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D010566:Virulence Factors, Bordetella
chemical_list: D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D006512:Hepatitis B Core Antigens; D010446:Peptide Fragments; D010566:Virulence Factors, Bordetella; C068929:pertactin