title: Antibodies to BrkA augment killing of Bordetella pertussis.
abstract: BrkA is a Bvg-regulated Bordetella pertussis protein that mediates serum resistance and adherence. It shares sequence identity with another B. pertussis virulence factor called pertactin, and it is a member of the diverse group of proteins found in Gram-negative bacteria that are secreted by an autotransporter mechanism. Sera, either from individuals who have been vaccinated with acellular pertussis vaccines, or from individuals who have no re-collection of recent infection with B. pertussis fail to kill wild-type B. pertussis, but kill brkA mutant strains very well. We examined whether BrkA could be neutralised in serum fitting this profile. BrkA is synthesised as a 103kDa precursor that is processed into a surface-associated N-terminal 73kDa passenger domain, and an outer-membrane embedded C-terminal 30kDa transporter moiety. Polyclonal antibodies were raised to a recombinant, re-folded histidine-tagged fusion protein representing the 73kDa passenger region. These anti-BrkA antibodies were shown to boost the existing bactericidal capacity of human serum against B. pertussis by neutralising BrkA.
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, #300-6174 University Boulevard, BC, V6T 1Z3, Vancouver, Canada.
authors.forename: D C
authors.initials: DC
authors.lastname: Oliver
authors.forename: R C
authors.initials: RC
authors.lastname: Fernandez
mesh_terms: D000328:Adult; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000918:Antibody Specificity; D000920:Antibody-Dependent Cell Cytotoxicity; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology* / Q000502:physiology; D001770:Blood Bactericidal Activity; D001886:Bordetella pertussis / Q000276:immunology*; D006801:Humans; D007106:Immune Sera; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000276:immunology; D010567:Pertussis Vaccine / Q000276:immunology; D011498:Protein Precursors / Q000276:immunology / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013047:Specific Pathogen-Free Organisms
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; C090007:BrkA protein, Bordetella pertussis; D007106:Immune Sera; D010446:Peptide Fragments; D010567:Pertussis Vaccine; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins