Hemagglutination activity of Treponema denticola grown in serum-free medium in continuous culture

Hemagglutination by different Treponema denticola strains was observed for erythrocytes of human, horse, bovine, and rabbit origin. The growth of T. denticola ATCC 33520 in serum-free medium in continuous culture enabled us to study the hemagglutinating activity of freshly harvested spirochetes of a defined physiological status. The hemagglutinating activity was cell bound and not related to motility or appendages, such as fimbriae. The activity was destroyed by proteolytic enzymes, heat, and alkylation, indicating that the agglutinin is of a proteinaceous nature. In addition, periodate oxidation of the spirochetes indicated the involvement of carbohydrate groups. Microscopic inspection of the hemagglutination mixtures at the titration endpoints revealed that only a part of the spirochete population was involved in the hemagglutination process. The hemagglutinating activity was found to be growth phase related. The activity was blocked by serum, while of all tested amino acids and carbohydrates, only sialic acid blocked the activity at low concentrations. In conclusion, we found a hemagglutinating activity in T. denticola which was cell bound and growth phase related. The agglutinin may be a glycoprotein, like lectin, that recognizes sialic acid as a receptor.