title: Glycolytic enzymes associated with the cell surface of Streptococcus pneumoniae are antigenic in humans and elicit protective immune responses in the mouse.
abstract: Streptococcus pneumoniae is a leading cause of otitis media, sinusitis, pneumonia, bacteraemia and meningitis worldwide. The drawbacks associated with the limited number of various capsular polysaccharides that can be included in the polysaccharide-based vaccines focuses much attention on pneumococcal proteins as vaccine candidates. We extracted an enriched cell wall fraction from S. pneumoniae WU2. Approximately 150 soluble proteins could be identified by 2D gel electrophoresis. The proteins were screened by 2D-Western blotting using sera that were obtained longitudinally from children attending day-care centres at 18, 30 and 42 months of age and sera from healthy adult volunteers. The proteins were further identified using matrix-assisted laser desorption ionization-time of flight mass spectrometry. Seventeen proteins were antigenic in children and adults, of which 13 showed an increasing antibody response with age in all eight children analysed. Two immunogenic proteins, fructose-bisphosphate aldolase (FBA) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and a control protein with known low immunogenicity, heat shock protein 70 (DnaK), were expressed in Escherichia coli, purified and used to immunize mice. Mouse antibodies elicited to the recombinant (r) FBA and rGAPDH were cross-reactive with several genetically unrelated strains of different serotypes and conferred protection to respiratory challenge with virulent pneumococci. In addition, the FBA used in this study (NP_345117) does not have a human ortholog and warrants further investigation as a candidate for a pneumococcal vaccine. In conclusion, the immunoproteomics based approach utilized in the present study appears to be a suitable tool for identification of novel S. pneumoniae vaccine candidates.
authors.affiliation: Pediatric Infectious Disease Unit, Soroka University Medical Center and the Department of Microbiology and Immunology, Ben Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: E
authors.initials: E
authors.lastname: Ling
authors.forename: G
authors.initials: G
authors.lastname: Feldman
authors.forename: M
authors.initials: M
authors.lastname: Portnoi
authors.forename: R
authors.initials: R
authors.lastname: Dagan
authors.forename: K
authors.initials: K
authors.lastname: Overweg
authors.forename: F
authors.initials: F
authors.lastname: Mulholland
authors.forename: V
authors.initials: V
authors.lastname: Chalifa-Caspi
authors.forename: J
authors.initials: J
authors.lastname: Wells
authors.forename: Y
authors.initials: Y
authors.lastname: Mizrachi-Nebenzahl
mesh_terms: D000328:Adult; D000367:Age Factors; D000818:Animals; D000917:Antibody Formation / Q000276:immunology*; D000942:Antigens, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D002462:Cell Membrane / Q000276:immunology; D002675:Child, Preschool; D029968:Escherichia coli Proteins / Q000276:immunology; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000276:immunology; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000276:immunology; D006019:Glycolysis / Q000276:immunology; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology; D006801:Humans; D007223:Infant; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology; D013296:Streptococcus pneumoniae / Q000201:enzymology / Q000276:immunology*
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); C063842:dnaK protein, E coli; D005634:Fructose-Bisphosphate Aldolase