title: A functional epitope of the pneumococcal surface adhesin A activates nasopharyngeal cells and increases bacterial internalization.
abstract: Pneumococcal surface adhesin A (PsaA) is a putative pneumococcal (Pnc) adhesin known to bind to nasopharyngeal (NP) epithelial cells. This study evaluated the effect of peptides within a functional domain of PsaA on NP cells. Detroit 562 NP cells were treated with synthetic peptides derived from PsaA (P4, P6, and P7; 28, 12, and 16 amino acids, respectively). The P4 peptide also binds to NP cells. Analysis of P4-treated NP cells by transmission electron microscopy revealed major cytological changes. Of 9 cytokines analyzed, a 6-fold increase in FGFb secretion at 3 and 6h (11-fold at 12h) was found post-P4 treatment of NP cells. There was a simultaneous reduction in the secreted levels of IL-6, IL-8, and VEGF. We observed enhancement in the adherence of Pnc strains to P4-treated NP cells (2-38-fold increase). Enhancement in adherence (2-fold increase) to P4-treated NP cells was also recorded with other streptococcal species (Streptococcus mitis and Streptococcus pyogenes). Internalization experiments demonstrated that 45% of the adherent bacteria were actually internalized after pretreatment with P4 peptide as compared to controls. Peptide fragments of P4, P6 and P7 did not activate NP cells to the extent of P4 peptide. The P4-mediated enhancement of Pnc adherence was blocked (100%) by anti-P4 antibodies, confirming the specificity of the P4 sequence for NP cell activation. Our data suggests that this functional domain of PsaA contained within the P4 sequence binds and activates NP cells to facilitate Pnc invasion.
authors.affiliation: Division of Bacterial Diseases, Centers for Disease Control and Prevention, Immunology Section, Building 18, Room B-105, MS A-36, 1600 Clifton Road, Atlanta, GA 30333, USA.
authors.forename: Gowrisankar
authors.initials: G
authors.lastname: Rajam
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Phillips
authors.forename: Elizabeth
authors.initials: E
authors.lastname: White
authors.forename: Julie
authors.initials: J
authors.lastname: Anderton
authors.forename: Craig W
authors.initials: CW
authors.lastname: Hooper
authors.forename: Jacquelyn S
authors.initials: JS
authors.lastname: Sampson
authors.forename: George M
authors.initials: GM
authors.lastname: Carlone
authors.forename: Edwin W
authors.initials: EW
authors.lastname: Ades
authors.forename: Sandra
authors.initials: S
authors.lastname: Romero-Steiner
mesh_terms: D018829:Adhesins, Bacterial / Q000494:pharmacology*; D000907:Antibodies, Bacterial / Q000276:immunology / Q000494:pharmacology; D001422:Bacterial Adhesion / Q000276:immunology*; D016207:Cytokines / Q000502:physiology; D004705:Endocytosis / Q000187:drug effects* / Q000502:physiology; D000939:Epitopes / Q000494:pharmacology*; D008074:Lipoproteins / Q000494:pharmacology; D046529:Microscopy, Electron, Transmission; D009305:Nasopharynx / Q000166:cytology* / Q000382:microbiology; D013296:Streptococcus pneumoniae / Q000737:chemistry*
chemical_list: D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D016207:Cytokines; D000939:Epitopes; D008074:Lipoproteins; C085000:PsaA protein, Streptococcus