Strains Of Streptococcus Sanguis Research Articles

Chemiluminescence and phagocytic reactions of human nuclear leukocytes incubated with Streptococcus sanguis

This experiment was designed to determine the role of indigenous organisms in gingival inflammation of the oral cavity, The test bacteria consisted of 3 strains of Streptococcus sanguis isolated from plaque at the gingival margin and grown in sure culture. These strains were subsequently designated as A, B and C. Peripheral nuclear leukocytes were isolated from 5 test subjects with clinically healthy gingiva. Luminol-dependent chemiluminescence and phagocytic functions of the 3 strains were measured, with the following results obtained. The mean peak value of luminol-dependent chemiluminescence following stimulation for strain A was 80.8 in the presence of serum, 38.8 in the presence of inactivated serum, and 20.5 in the absence of serum. For strain B it was 82.3 in the presence of serum, 6.4 in the presence of inactivated serum, and 2.5 in the absence of serum. For strain C it was 68.1 in the presence of serum, 65.3 in the presence of inactivated serum, and 8.3 in the absence of serum. In the absence of plasma strains A and B showed slight phagocytosis. In strain C, however, no phagocytosis was noted. In the presence of inactivated serum phagocytosis was increased strains A and B, but no phagocytosis was noted in strain C. In the presence of blood. phagocytosis was increased in strains A and B, but no phagocytosis was noted in strain C.

Roxithromycin as a possible agent for prophylaxis of endocarditis: a study in normal volunteers.

A single dose of roxithromycin, 300 mg, was taken by six healthy male volunteers on three occasions at consecutive weekly intervals. It was well tolerated. On the first two occasions, roxithromycin was assayed in serum samples taken at intervals up to 8 h after the administration. The mean peak concentration at 1 h was 3.0 mg/l (range 0.3-7.3). The serum samples from the volunteers showed variable bactericidal activity against a strain of Streptococcus sanguis isolated from a case of bacterial endocarditis. Roxithromycin was not detected in saliva or gingival fluid. Smooth surface plaque samples taken at intervals were investigated for the emergence of streptococci resistant to roxithromycin at 2 and 8 mg/l. Initially two volunteers had small number of roxithromycin-resistant streptococci. At the end of the study all six volunteers had resistant streptococci detectable in their plaque samples and these accounted for 100% of the streptococci in two volunteers. The most resistant isolates (in several cases with MIC greater than 64 mg/l) were Str. sanguis or Str. mitior; individual volunteers tended to yield the same strain on consecutive samplings.

Adherence of Streptococci to Surface-modified Glass

Four types of surface-modified glass were prepared. Aminopropyl glass was prepared by alkylsilylation of glass slides with gamma-aminopropyltriethoxysilane. This glass carries primary amino groups which may be protonated at pH 7.2. Owing to the presence of both positively charged ions and hydrophobic ethoxyl groups, the glass is considered to be amphipathic. Three other types of surface-modified glass slides were prepared from aminopropyl glass by forming Schiff's bases with three aldehydes: glucose, glyoxylic acid and hexanal. The aldehyde-treated slides were subsequently reduced using sodium borohydride. Thus, the surface of the glass was rendered hydrophilic, ampholytic or hydrophobic, respectively. The adherence of two Streptococcus sanguis strains and two Streptococcus mutans strains to the surface-modified glass slides was studied. Different strains showed differences in adherence to these slides depending on their physico-chemical surface properties. For S. sanguis ATCC 10556, hydrophobic bonds seemed to be most important, while in S. mutans OMZ 176, ionic interactions made the highest contribution to adhesion. Hydrogen bonds seemed to contribute least to adherence.

Cloning of a Streptococcus sanguis adhesin which mediates binding to saliva-coated hydroxyapatite.

Chromosomal DNA from a salivary aggregating strain of Streptococcus sanguis 12 was partially digested with PstI and ligated into the plasmid vector pUC18 and transformed into Escherichia coli JM83. A total of 1,700 recombinant clones of E. coli were examined by a colony immunoassay with antisera raised against either S. sanguis 12 whole cells or S. sanguis 12 surface fibrils. Five clones which reacted with one or the other antiserum were shown to be unique by Western blotting (immunoblotting) and restriction endonuclease digestion. One recombinant plasmid pSA2 expressed two proteins with Mrs of 20,000 and 36,000. The 36,000-Mr protein has been designated SsaB. Both proteins were purified to homogeneity by Sephadex G-75 and ion-exchange chromatography. The proteins were present in mutanolysin digests of whole-cell lysates of S. sanguis 12 and in the non-saliva-aggregating variant 12na and the hydrophilic variant 12L. Polyclonal antiserum raised against the SsaB protein reacted strongly with the cell surfaces of S. sanguis 12 and 12na but not with that of 12L. SsaB inhibited the adhesion of S. sanguis 12na to saliva-coated hydroxyapatite, indicating that the adhesin mediates the binding to the pH-sensitive receptor.

Response of a Streptococcus sanguis strain to arginine-containing peptides.

For dental plaque organisms such as Streptococcus sanguis, the ecological importance of the ability to utilize arginine as an energy source has been established in previous studies. The present investigation was undertaken to determine the ability of a strain of S. sanguis to process unsubstituted arginine-containing peptides. The organism was grown under glucose-limited conditions in a chemically defined medium, and peptide was added to washed, resting cells in a pH-stat at pH 7.0. Filtrates taken at appropriate time intervals were assayed for peptide, free amino acids, and metabolites. Irrespective of the position of the arginine residue, all peptides tested were attacked, although those that possessed a C-terminal arginine (including a tetrapeptide) were processed at a faster rate than were those in which arginine was N terminal. However, C-terminal arginine was cleaved only slowly from a peptide containing 24 residues. In each case, most of the released arginine was converted to ornithine via the arginine deiminase pathway. Such peptidase activities appeared to occur at or near the cell surface and were probably constitutive. It was found that the organism grew in chemically defined medium containing arginine that was present solely in the form of a tripeptide, and also that a strain of S. mutans possessed only a limited ability to attack arginine-containing peptides and was unable to utilize the released arginine.

Determination of the cell surface hydrophobicity of oral bacteria using a modified hydrocarbon adherence method

Hydrophobic interactions between bacterial cell surfaces and colonisable substrates have been implicated in the mechanisms of bacterial adherence. However, current methods of assessing bacterial hydrophobicity as a function of adherence to liquid hydrocarbons (especially hexadecane) do not always produce accurate or reproducible results. Therefore, the present technique was developed using xylene. The hydrophobic surface properties of fresh and type strains of Bacteriodes gingivalis, Bacteriodes intermedius, Capnocytophaga spp., Streptococcus salivarius and Streptococcus sanguis suspended either in saliva ions buffer (SIB) or in saliva diluted in SIB were measured. In SIB the test strains were predominantly hydrophobic. The addition of saliva caused a significant reduction (P < 0.05) in hydrophobicity compared to SIB alone, with 80% of the strains tested. Since oral bacteria will be suspended in saliva in vivo, it is concluded that bacteria in the oral cavity may be less hydrophobic than previous studies have suggested.

Transformation ofStreptococcus sanguisChallis with a plasmid ofStreptococcus pneumoniae

Abstract The present work is concerned with plasmid transformation of Streptococcus sanguis strain Challis with derivatives of pDP1/pSMB1, the only plasmid found to occur naturally in Streptococcus pneumoniae . Two recombinant plasmids derived from the cryptic pSMB1 were used: pDP27 (4.5 kb) conferring resistance to chloramphenicol (Cm), and pDP28 (7.8 kb), a shuttle plasmid, conferring resistance to Cm in Escherichia coli , and resistance to erythromycin (Em) in pneumococcus. It could be shown that pSMB1 can replicate in S. sanguis ; in fact, Challis strain V288 was transformed to Cm-resistance and to Em-resistance by pDP27 and pDP28 respectively. Shuttle plasmid pDP28 can transform S. sanguis both when isolated from pneumococcus and from E. coli , albeit with a different efficiency. The low frequency of transformation observed when pDP28 was isolated from E. coli DH1 ( recA ) was shown to be due to lack of multimeric forms of the plasmid in the DNA preparations obtained from this strain. When pDP28 was isolated from E. coli C600 (RecA + ), multimeric forms were present, and transformations of S. sanguis was more efficiency Using pDP28 as vector in cloning experiments, where S. sanguis was the host of the recombinant DNA molecules, treatment of the vector with alkaline phosphatase inhibited the recovery of recombinant clones.

Transformation of Streptococcus sanguis Challis with a plasmid of Streptococcus pneumoniae

The present work is concerned with plasmid transformation of Streptococcus sanguis strain Challis with derivatives of pDP1/pSMB1, the only plasmid found to occur naturally in Streptococcus pneumoniae. Two recombinant plasmids derived from the cryptic pSMB1 were used: pDP27 (4.5 kb) conferring resistance to chloramphenicol (Cm), and pDP28 (7.8 kb), a shuttle plasmid, conferring resistance to Cm in Escherichia coli, and resistance to erythromycin (Em) in pneumococcus. It could be shown that pSMB1 can replicate in S. sanguis; in fact, Challis strain V288 was transformed to Cm-resistance and to Em-resistance by pDP27 and pDP28 respectively. Shuttle plasmid pDP28 can transform S. sanguis both when isolated from pneumococcus and from E. coli, albeit with a different efficiency. The low frequency of transformation observed when pDP28 was isolated from E. coli DH1 (recA) was shown to be due to lack of multimeric forms of the plasmid in the DNA preparations obtained from this strain. When pDP28 was isolated from E. coli C600 (RecA+), multimeric forms were present, and transformations of S. sanguis was more efficiency Using pDP28 as vector in cloning experiments, where S. sanguis was the host of the recombinant DNA molecules, treatment of the vector with alkaline phosphatase inhibited the recovery of recombinant clones.

Environmental pH as a factor in the competition between strains of the oral streptococci Streptococcus mutans, S. sanguis, and "S. mitior" growing in continuous culture.

Strains of Streptococcus mutans (biotype 1), Streptococcus sanguis, and Streptococcus mitior have been grown in mixed continuous culture in a semidefined medium under glucose limitation at a growth rate of D = 0.1 h-1. The effect of varying the environmental pH on the proportions of the different populations within the community has been determined. Initially the populations were allowed to reach steady state at pH 7.0 when S. sanguis was dominant with S. mutans and "S. mitior" maintaining similar populations. The medium pH was then lowered in steps of 0.5 pH units from pH 7.0 to 4.5, and the community was grown at each step for at least 15 generations. Viable counts of each species were made at 24-h intervals. The population ratios established at pH 7.0 remained relatively stable when the environmental pH was set at pH 6.5. However, after the medium pH was lowered to 6.0 (days 18-27), the population of S. mutans began to increase and the S. mitior population began to decline. A further change was seen at pH 5.5 (days 27-34) when S. mutans became dominant, S. sanguis declined, and S. mitior was not detectable. At pH 4.5, both S. mutans and S. sanguis were reduced in numbers, but survived until the experimental run was terminated (44 days). Samples of culture fluid were taken throughout the experiment and analyzed for the presence of the acid products of glucose metabolism. The amounts of lactic acid produced by the community increased as the environmental pH was lowered.(ABSTRACT TRUNCATED AT 250 WORDS)

Serological relationships between serotype-III Streptococcus sanguis and Lancefield group-H Streptococci

Certain strains of Streptococcus sanguis and group-H streptococci have been shown to have similar physiological properties and serological specificities. Serological studies revealed that serotype-III S. sanguis shared a common antigen with the so-called "British" group-H streptococci, but not with the "American" group-H streptococci. Serotype-III antigen was extracted in cold 5% trichloroacetic acid from isolated cell walls of S. sanguis ATCC 10558, and purified chromatographically. The purified serotype-III antigen consisted of neutral and amino sugars and some phosphorus, and was negatively charged. Hapten inhibition of the quantitative precipitin reaction between serotype-III antigen and antiserum indicated the strong possibility of alpha-glucosidic linkages being an immunodeterminant of the antigen.

A bacteriocin of Actinobacillus actinomycetemcomitans.

An inhibitory factor from Actinobacillus actinomycetemcomitans Y4 was isolated, and its properties indicated that it was a bacteriocin (actinobacillicin). The bacteriocin was active against Streptococcus sanguis strains, Streptococcus uberis (FDC1), and Actinomyces viscosus T14 as well as other strains of A. actinomycetemcomitans, but not against other crevicular bacteria, including other streptococci and actinomycetes. The activity of this bacteriocin was inhibited by pronase, trypsin, and heat (45 min at 56 degrees C) but not by DNase, RNase, phospholipase, exposure to UV light, or low pH (1.0 to 6.5). Although actinobacillicin markedly inhibited glycolysis in S. sanguis, the primary mechanism of its bactericidal action appears to be alterations in cell permeability, with the resultant leakage of RNA, DNA, and other essential intracellular macromolecules. These findings provide an ecologic explanation for the reciprocal growth relationship between A. actinomycetemcomitans and S. sanguis/Actinomyces viscosus observed in localized juvenile periodontitis.

Responses of Platelets to Strains of Streptococcus sanguis: Findings in Healthy Subjects, Bernard-Soulier, Glanzmann’s, and Collagen-Unresponsive Patients

The ability of endocarditis and dental strains of Streptococcus sanguis to induce platelet aggregation in plasma (PRP) from normal subjects were examined and compared to responses of PRP with known platelet membrane glycoprotein (GP) and response defects. S. sanguis strains differed in their ability to induce normal PRPs to aggregate. Strains that induced PRP aggregation in more than 60% of donors were significantly faster agonists (mean lag times to onset of aggregation less than 6 min) than those strains inducing response in PRPs of fewer than 60% of donors. Platelets from patients with Bernard-Soulier syndrome aggregated in response to strains of S. sanguis. In contrast, platelets from patients with Glanzmann's thrombasthenia and from a patient with a specific defect in response to collagen were unresponsive to S. sanguis. These observations show that GPIb and V are not essential, but GPIIb-IIIa and GPIa are important in the platelet response mechanism to S. sanguis. Indeed, the data suggests that the platelet interaction mechanisms of S. sanguis and collagen may be similar.

A simplified diagnostic system for cultural detection and enumeration of Streptococcus mutans.

A simple dip-slide test (Cariescreen SM) based on MSB selective agar was devised for detection and quantitation of Streptococcus mutans in oral samples. For this test, a bacitracin tablet is dissolved in a vial containing buffered saline diluent. Paraffin-stimulated saliva is collected in this diluent vial. A dip slide which incorporates a modified MSB agar (minus bacitracin) is immersed briefly in the diluted saliva. After addition of a CO2-generating tablet, the screw-cap dip slide is closed tightly in the vial and incubated for two days at 37 degrees C and one day at room temperature. S. mutans populations in saliva are estimated by comparison with a colony density chart. Growth of reference strains of S. mutans was equivalent on Cariescreen SM dip slides and on MSB agar plates. Reference strains of Streptococcus sanguis, Streptococcus salivarius, Streptococcus mitis, and Streptococcus milleri did not grow on Cariescreen SM dip slides. Aliquots of saliva from 50 schoolchildren and 51 adults were tested by the dip-slide method and by conventional plating methods in MSB agar. Very good correlation (0.93) between the two methods was obtained. This simplified S. mutans detection system is suitable for use by clinical personnel in dental clinics or other non-laboratory settings for identification of subjects potentially at risk for caries.

Characterization of monoclonal antibodies specific for adhesion: isolation of an adhesin of Streptococcus sanguis FW213.

Monoclonal antibodies reactive to an adhesive strain of Streptococcus sanguis (FW213) and nonreactive to a nonadhesive mutant (JL7) were derived from the fusion of myeloma line X63Ag8.653 and spleen cells from BALB/c mice immunized with live S. sanguis cells. Five cell lines, belonging to subclasses of immunoglobulin G, produced monoclonal antibodies specifically directed against the adhesive strain. All five antibodies also failed to react with five additional, independently isolated, nonadhesive mutants. A spontaneous mutant of FW213 (VT508) that no longer reacted with monoclonal antibody F51 (MAbF51) was isolated by serial agglutination with the antibody. Langmuir adsorption isotherms of VT508 indicated that this mutant also had altered ability to adhere to saliva-coated hydroxyapatite further confirming the specificity of MAbF51 for adhesion. Electron microscopy revealed that VT508 had lost the peritrichous fimbriae associated with the adhesion of FW213. MAbF51 was used to purify the adhesin from lysozyme cell extracts by using an affinity column of MAbF51 linked to Sephacryl S1000. Purity was suggested by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and crossed immunoelectrophoresis (CIEP). The adhesin had a molecular weight greater than 150,000 and was not denatured in sodium dodecyl sulfate reducing gels. Two peaks of near electrophoretic mobility were detected in CIEP when the purified material was run against polyclonal antibody to the whole cell. Tandem CIEP analysis and immunoprecipitation provided evidence that the two peaks represented the same antigen in two different forms.

Coagglutination reactions between Candida albicans and oral bacteria.

An agglutination assay for detecting intermicrobial adherence between the cells of Candida albicans and various oral bacteria is described. Strains of Streptococcus sanguis, S. salivarius, S. mutans, S. mitis, Fusobacterium nucleatum and Actinomyces viscosus all coagglutinated with C. albicans. No interaction could be demonstrated between the cells of Bacteroides melaninogenicus and those of C. albicans. Preliminary investigations of these interactions suggest that binding of F. nucleatum and A. viscosus to C. albicans is mediated by bacterial proteins, possibly lectins. Other mechanisms must account for the binding of oral streptococci to C. albicans. The possible implications of these findings in relation to oral mucosal colonisation and oral candidal clearance are discussed.

Preparation of a sialic acid-binding protein from Streptococcus mitis KS32AR.

A recent report has identified a lectin on the surfaces of several strains of Streptococcus mitis and Streptococcus sanguis with specificity for an N-acetylneuraminic acid alpha 2,3-galactose-beta 1,3-N-acetylgalactosamine sequence (P.A. Murray, M.J. Levine, L.A. Tabak, and M.S. Reddy, Biochem. Biophys. Res. Commun. 106:390-396, 1982). In the present study, purification and characterization of this sialic acid-binding protein (SABP) was begun. A clinical isolate of S. mitis was grown to mid stationary phase in synthetic FMC medium and then extracted with lithium 3,5-diiodosalicylate. Lyophilized extract was subjected to gel filtration on a Sephadex G-200 column, giving four protein peaks (A to D). Peak B, shown by hemagglutination assay to contain SABP, was next subjected to affinity chromatography on a Sepharose-4B matrix coupled to fetuin glycopeptides. After an extensive washing, peak B materials bound to the affinity matrix were eluted with buffered N-acetylneuraminic acid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis with 2-mercaptoethanol on 7.5% gels of affinity-purified materials revealed components of 96, 70, and 65 kilodaltons (kDa). Without reducing agent, only the 65-kDa band and materials which did not penetrate the gel were visualized, suggesting that the 96- and 70-kDa components were disulfide linked. The chemical cross-linking agent, disuccinimidyl suberate, was used to demonstrate specific interactions between the SABP preparation and [14C]fetuin glycopeptides. After cross-linking, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography revealed the 96- and 70-kDa components, indicating that the SABP is at least bivalent. These findings support our previous suggestion that human salivary glycoproteins facilitate clearance of selected oral streptococci via specific interactions between sialic acid-containing oligosaccharides and a carbohydrate-binding protein on the bacterial cell surface.

Calcium-dependent salivary agglutinin with reactivity to various oral bacterial species.

Human parotid agglutinins from three individuals were isolated by adsorption to and desorption from strains of Streptococcus mutans belonging to serotypes a, b, c, d, and e and strains of Lactobacillus casei, Actinomyces viscosus, and Streptococcus sanguis. The desorption was achieved by suspending centrifuged saliva-coated microorganisms in 10 mM phosphate buffer (pH 6.8) containing 0.154 M sodium chloride. After another centrifugation, agglutinin activity was recovered in the supernatants. The L. casei strain was not agglutinated by any of the agglutinin extracts or by saliva, but all the other strains were agglutinated to a variable extent. However, all strains, including the nonagglutinating L. casei strain, adsorbed and desorbed agglutinins active for other strains. The agglutinin extracts from S. mutans serotype c, S. sanguis, and A. viscosus were purified and characterized by electrophoretic and immunological techniques. The purified preparations were positively stained for protein and carbohydrate, and the molecular weights were estimated to be 440,000. All agglutinin extracts needed calcium in the range of 0.1 to 0.5 mM to be active, and for a single strain, all agglutinins gave the same degree of agglutination, indicating that the isolated agglutinins may be of the same molecular species, a hypothesis that was also confirmed by the preliminary characterization of the purified agglutinins. This type of agglutinin, which seems to exert its activity among various bacterial species, could be important in mediating bacterial coaggregation and thus may add to the effect of specific agglutinins in the clearance of bacteria from the human mouth.

Variable colonization by oral streptococci in molar fissures of monoinfected gnotobiotic rats.

Germfree Sprague-Dawley rats, fed a high-sucrose diet, were monoinfected with strains of Streptococcus mitis, Streptococcus sanguis, and Streptococcus mutans. Viable cell recoveries from six molar teeth, considered to reflect mainly bacterial colonization of intact fissures, were in the order of 10(6), 10(7), and 10(8) CFU, respectively. Some of the implications of the variation of bacterial plaque-forming ability in the rat model are discussed.

Adherence of Streptococcus sanguis to hydroxyapatite coated with lysozyme and lysozyme-supplemented saliva

The adherence of [3H]thymidine-labeled Streptococcus sanguis strains to bare hydroxyapatite and to hydroxyapatite coated with a range of concentrations of lysozyme, poly-L-lysine, poly-L-glutamic acid, whole saliva supernatant, and combinations of some of the above was studied. Adherence of several strains of S. sanguis to bare hydroxyapatite and saliva-coated hydroxyapatite was compared. Saliva present as a pellicle on the hydroxyapatite inhibited adherence of some strains (903, M-5, 73X11) and stimulated that of others (S35, B-4, 66X49). Strains 903 and S35 were chosen for further study. Adherence of both strains was stimulated up to fivefold by the presence of adsorbed lysozyme or poly-L-lysine on the hydroxyapatite, whereas poly-L-glutamic acid inhibited adherence (80 to 95%). Adherence of strain S35 to hydroxyapatite coated with combinations of saliva and (i) lysozyme, (ii) poly-L-lysine, or (iii) poly-L-glutamic acid was unaffected compared with adherence to hydroxyapatite coated with saliva alone. In contrast, adherence of strain 903 to hydroxyapatite coated with combinations of saliva and either lysozyme or poly-L-lysine was inhibited up to ca. 90% compared with hydroxyapatite coated with saliva alone. Strain 903 was also unaffected by combinations of poly-L-glutamic acid and saliva on the hydroxyapatite. Adherent cells of both strains were completely (greater than 90%) eluted with high-ionic-strength buffer from either bare hydroxyapatite or hydroxyapatite coated with lysozyme alone. Adherent cells of strain S35 were only poorly eluted (25%) from hydroxyapatite coated with either saliva alone or saliva and lysozyme. Strain 903 elution from hydroxyapatite coated with either saliva alone or saliva and lysozyme was essentially complete. These observations were taken to indicate that the two test strains adhered to saliva-coated hydroxyapatite by different mechanisms. Protein-coated hydroxyapatite was shown not to be saturated under the conditions described here. Examination by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the variously supplemented salivary pellicles formed on the hydroxyapatite demonstrated that major changes in salivary protein composition did not occur when lysozyme, poly-L-lysine, or poly-L-glutamic acid was used to supplement saliva. Lysozyme-dependent aggregation of strain 903 was shown not to occur under the conditions of our experiments. We suggest that the basis for stimulation of adherence to hydroxyapatite coated only with lysozyme is an increase in the cationic surface area available for electrostatic adherence of the microorganisms.(ABSTRACT TRUNCATED AT 400 WORDS)

Isolation and characterization of a 60-kilodalton salivary glycoprotein with agglutinating activity against strains of Streptococcus mutans.

A bacterial agglutinin specific for strains of Streptococcus mutans was isolated from human saliva. Physiochemical analyses showed the agglutinin to be a glycoprotein with a molecular weight of 60,000. The agglutinin aggregated four of the eight strains of Streptococcus mutans tested but did not aggregate the strains of Streptococcus salivarius, Streptococcus sanguis, and Streptococcus mitis tested. Chemical modification of carbohydrate moieties of the agglutinin with sodium metaperiodate had no effect on aggregation, whereas modification of the polypeptide portion with trypsin abolished aggregating activity. A set of five murine hybridoma antibodies was employed to further analyze the agglutinin. Two carbohydrate-specific antibodies, directed against D-mannose and N-acetylgalactosamine moieties, respectively, failed to block agglutinin- or whole saliva-mediated aggregation of S. mutans cells. In contrast, two antibodies directed against pronase-sensitive antigenic sites blocked both agglutinin- and saliva-mediated aggregation of S. mutans cells. Western blot analysis with the agglutinin-specific hybridoma antibodies demonstrated the agglutinin in whole saliva and in artificial tooth pellicles formed on hydroxyapatite beads incubated with saliva. These results suggest that a 60-kilodalton glycoprotein of human saliva is a bacterial agglutinin with specificity for certain strains of S. mutans. They further suggest that aggregation is mediated by polypeptide rather than carbohydrate determinants of the glycoprotein.