Many researchers have suggested that the role of glucan-mediated interactions in the adherence of Streptococcus mutans is restricted to accumulation of this cariogenic bacterium following its sucrose (i.e., glucan)-independent binding to saliva-coated tooth surfaces. However, the presence of enzymatically active glucosyltransferase in salivary pellicle suggests that glucans could also promote the initial adherence of S. mutans to the teeth. In the present study, the commonly used hydroxyapatite adherence assay was modified to include the incorporation of glucosyltransferase and the synthesis of glucans in situ on saliva-coated hydroxyapatite beads. Several laboratory strains and clinical isolates of S. mutans were examined for their ability to adhere to experimental pellicles, either with or without the prior formation of glucans in situ. Results showed that most strains of S. mutans bound stereospecifically to glucans synthesized in pellicle. Inhibition studies with various polysaccharides and fungal dextranase indicated that alpha 1,6-linked glucose residues were of primary importance in the glucan binding observed. Scanning electron microscopic analysis showed direct binding of S. mutans to hydroxyapatite surface-associated polysaccharide and revealed no evidence of trapping or cell-to-cell binding. S. mutans strains also attached to host-derived structures in experimental pellicles, and the data suggest that the bacterial adhesins which recognize salivary binding sites were distinct from glucan-binding adhesins. Furthermore, glucans formed in experimental pellicles appeared to mask the host-derived components. These results support the concept that glucans synthesized in salivary pellicle can promote the selective adherence of the cariogenic streptococci which colonize human teeth.

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This study aimed to determine physical and kinetic properties ofglucosyltransferase (GTF) adsorbed onto hydroxyapatite (HA) surfaces.For development of a solid-phase enzyme assay, 4.0-mg samples of washed HA powder were exposed to centrifuged whole saliva (WSHA) or buffer, and subsequently exposed to a GTF solution. The activities of GTF adsorbed to H4 and that remaining in solution were measured. WSH4 was more effective in adsorbing GTF than was naked HA. Enzyme activity on the surface of WSHA was enhanced; more activity was detected on WSHA than was apparently removed from solution. A similar effect was observed when GTF was adsorbed to naked HA from a mixture with lysozyme or saliva; however, no enhancement was seen when GTF was adsorbed from a mixture with albumin. Compared with GTF in solution, adsorbed GTF displayed activity over a much wider range ofpH values. Temperature-activity profiles indicated that GTF adsorbed to surfaces had a lower temperature optimum (400C) than did soluble enzyme (45'C), and that the bound enzyme was more resistant to adverse effects of heat at elevated temperatures. The majority of glucan made by GTF adsorbed to parotid saliva-coated HA remained attached to the surface. The activity of lysozyme adsorbed to HA was reduced by adsorption of GTF to the same surface and was almost completely abolished by formation of glucans by the adsorbed GTF. These results suggest that soluble bacterial enzymes found in saliva can be incorporated into pellicle, interact with host-derived molecules on the surfaces of teeth, express enzymatic activity, and potentially influence the biological properties ofpellicle.

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Certain strains of Streptococcus sanguis adhere (Adh+) selectively to human platelets and, in plasma, induce them to aggregate (Agg+) into in vitro thrombi. In this study, we examined 18 recent endocarditis and dental plaque isolates of microorganisms that were biotyped as S. sanguis for coexpression of platelet interactivity phenotypes with another possible virulence factor in bacterial endocarditis, dextran synthesis. Detectable production of extracellular glucosyltransferase ranged from 0.2 to 66 mU/mg of culture fluid for 10 representative strains tested. Production of extracellular or cell-associated glucosyltransferase, fructosyltransferase, and soluble or insoluble dextrans was not necessarily coexpressed with platelet interactivity phenotypes, since the levels of production of soluble and insoluble dextrans varied among representative Adh+ Agg+ and Adh-Aggstrains. Analysis of a second panel of 38 fresh dental plaque isolates showed that S. sanguis distributes in a reproducible manner into the possible phenotype groups. Strains with different platelet interactivity phenotypes were distinguished with a panel of four murine monoclonal antibodies (MAbs) raised against Adh+ Agg+ strain 133-79 and screened to rule out artifactual reactions with antigenic components in culture media. The MAbs reacted selectively with Adh+ Agg+ strains in a direct-binding, whole-cell, enzyme-linked immunosorbent assay and also inhibited their interactions with platelets. Analysis of minimal tryptic digests of many strains, including variants that failed to bind the MAbs, suggested that some noninteractivity phenotypes possess cryptic surface determinants. Since the ability to adhere to platelets and induce them to aggregate is relatively stable, these traits may be useful in a phenotyping scheme for these Lancefield nontypeable streptococci.