title: Cross-reactivity of antipneumococcal surface protein C (PspC) antibodies with different strains and evaluation of inhibition of human complement factor H and secretory IgA binding via PspC.
abstract: Pneumococcal surface protein C (PspC) is an important candidate for a cost-effective vaccine with broad coverage against pneumococcal diseases. Previous studies have shown that Streptococcus pneumoniae is able to bind to both human factor H (FH), an inhibitor of complement alternative pathway, and human secretory IgA (sIgA) via PspC. PspC was classified into 11 groups based on variations of the gene. In this work, we used three PspC fragments from different groups (PspC3, PspC5, and PspC8) to immunize mice for the production of antibodies. Immunization with PspC3 induced antibodies that recognized the majority of the clinical isolates as analyzed by Western blotting of whole-cell extracts and flow cytometry of intact bacteria, while anti-PspC5 antibodies showed cross-reactivity with the paralogue pneumococcal surface protein A (PspA), and anti-PspC8 antibodies reacted only with the PspC8-expressing strain. Most of the isolates tested showed strong binding to FH and weaker interaction with sIgA. Preincubation with anti-PspC3 and anti-PspC5 IgG led to some inhibition of binding of FH, and preincubation with anti-PspC3 partially inhibited sIgA binding in Western blotting. The analysis of intact bacteria through flow cytometry showed only a small decrease in FH binding after incubation of strain D39 with anti-PspC3 IgG, and one clinical isolate showed inhibition of sIgA binding by anti-PspC3 IgG. We conclude that although anti-PspC3 antibodies were able to recognize PspC variants from the majority of the strains tested, partial inhibition of FH and sIgA binding through anti-PspC3 antibodies in vitro could be observed for only a restricted number of isolates.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, Brazil.
authors.forename: Adriana T
authors.initials: AT
authors.lastname: Moreno
authors.forename: Maria Leonor S
authors.initials: ML
authors.lastname: Oliveira
authors.forename: Paulo L
authors.initials: PL
authors.lastname: Ho
authors.forename: Cintia F M
authors.initials: CF
authors.lastname: Vadesilho
authors.forename: Giovana M P
authors.initials: GM
authors.lastname: Palma
authors.forename: Jorge M C
authors.initials: JM
authors.lastname: Ferreira
authors.forename: Daniela M
authors.initials: DM
authors.lastname: Ferreira
authors.forename: Silvia R
authors.initials: SR
authors.lastname: Santos
authors.forename: Marina B
authors.initials: MB
authors.lastname: Martinez
authors.forename: Eliane N
authors.initials: EN
authors.lastname: Miyaji
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D015153:Blotting, Western; D017242:Complement Factor H / Q000037:antagonists & inhibitors; D003429:Cross Reactions; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D007071:Immunoglobulin A, Secretory / Q000097:blood / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013296:Streptococcus pneumoniae / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; C493495:CFH protein, human; D004269:DNA, Bacterial; D007071:Immunoglobulin A, Secretory; D007074:Immunoglobulin G; C110344:SpsA protein, Streptococcus pneumoniae; D017242:Complement Factor H