title: Characterization of two distinct opsonic and protective epitopes within the alpha C protein of the group B Streptococcus.
abstract: Group B Streptococcus (GBS) is a major cause of neonatal sepsis, meningitis in early infancy, postpartum endometritis, and serious invasive infections in adults in the United States. We previously cloned, sequenced, and characterized the alpha antigen gene, bca, and showed that the alpha C protein of GBS is a trypsin-resistant, surface-associated polypeptide that contains a signal sequence, a unique N terminus, nine identical tandem repeats, and a C-terminal membrane anchor structure. Polyclonal antiserum raised to the recombinant alpha C protein and an opsonic monoclonal antibody, 4G8, raised to the native protein from GBS have been shown to be protective in a mouse model. The binding site of 4G8 has now been localized to the tandem repeat region of the alpha C protein. To determine whether the N terminus of the alpha C protein contains additional opsonic and/or protective epitopes, the sequence corresponding to the alpha C protein N terminus was subcloned into a pET vector, the expressed peptide from Escherichia coli was purified by Ni2+ affinity chromatography, and rabbit polyclonal antibodies were raised to the purified recombinant peptide. Antibodies to the alpha C protein N terminus were shown to be opsonic by an in vitro opsonophagocytosis assay. In addition, 69% of newborn mouse pups from mothers passively immunized with the antiserum to the recombinant N-terminal polypeptide of the alpha C protein were protected against lethal challenge with GBS A909. These data indicate that at least two distinct regions of the alpha C protein, the N terminus and the tandem repeat region, contain opsonic and protective epitopes.
authors.affiliation: Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, USA.
authors.forename: D E
authors.initials: DE
authors.lastname: Kling
authors.forename: C
authors.initials: C
authors.lastname: Gravekamp
authors.forename: L C
authors.initials: LC
authors.lastname: Madoff
authors.forename: J L
authors.initials: JL
authors.lastname: Michel
mesh_terms: D000328:Adult; D000818:Animals; D000954:Antigens, Surface / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D018604:Epitope Mapping; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology; D051379:Mice; D011817:Rabbits; D013292:Streptococcus agalactiae / Q000276:immunology*
chemical_list: D000954:Antigens, Surface; D001426:Bacterial Proteins; D016056:Immunodominant Epitopes; C077961:alpha C protein, group B streptococci