Characterization of monoclonal antibodies to fimbria-associated adhesins of Bacteroides loescheii PK1295.
Bacteroides loescheii PK1295 fimbriae, which mediate the lactose-sensitive coaggregation with Streptococcus sanguis 34 and the lactose-insensitive coaggregation with Actinomyces israelii PK14, were injected into mice to raise adhesin-specific monoclonal antibodies (MAbs). Supernatants of hybridomas were screened for the capacity to inhibit coaggregation and agglutinate intact bacteria. Of the 10 MAbs that were isolated, 4 were specific and potent inhibitors of the coaggregation between B. loescheii and S. sanguis and two other MAbs specifically inhibited the B. loescheii-A. israelii interaction. None of the six MAbs which inhibited adherence were capable of agglutinating whole cells of B. loescheii, whereas the four remaining MAbs agglutinated whole cells but had no effect on coaggregation. Fab fragments of two MAbs, one that inhibited the coaggregation with S. sanguis and another that inhibited the interaction with A. israelii, also were shown to inhibit the respective coaggregation interactions, suggesting that each of the immunoglobulins recognized its adhesin molecule at or near the active sites. By immunoblotting or immunoprecipitation, the S. sanguis adhesin-specific MAbs reacted with a 75-kilodalton polypeptide present in fimbria-enriched preparations, whereas the A. israelii adhesin-specific MAbs recognized a 45-kilodalton polypeptide in the same preparations. By screening hybridoma supernatants directly for their capacity to block coaggregation, we isolated MAbs which were used to establish that the B. loescheii-S. sanguis and the B. loescheii-A. israelii interactions were mediated by different adhesins.
D000190:Actinomyces / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000502:physiology; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000276:immunology*; D001439:Bacteroides / Q000276:immunology*; D001666:Binding Sites, Antibody; D005260:Female; D010861:Fimbriae, Bacterial / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D013298:Streptococcus sanguis / Q000276:immunology
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins