title: The effect of extracellular polysaccharides from Streptococcus mutans on the bactericidal activity of human neutrophils.
abstract: Extracellular polysaccharides (PS) synthesized by oral bacteria constitute one of their major virulence factors. The PS, synthesized from sucrose, facilitate adhesion and colonization by bacteria to tooth surfaces. The study was designed to test the effect of in situ production of extracellular PS by Streptococcus mutans on the bactericidal activity of human neutrophils. These effects were tested on bacteria pre-exposed to sucrose (PS-positive Strep. mutans) and compared to bacteria not exposed to sucrose (PS-negative Strep. mutans). The interactions between neutrophils and Strep. mutans were tested in suspension and on bacteria in an experimental model of dental plaque. Viability of Strep. mutans was measured by [3H]-thymidine incorporation into the bacteria. Degranulation of neutrophils was evaluated by the release of lysozyme, and the production of reactive oxygen products was measured by chemiluminescence. When neutrophils were incubated with suspended bacteria, the viability of PS-negative Strep. mutans was 20% of that of bacteria not incubated with neutrophils (control), while the viability of PS-positive Strep. mutans was 40% of the control. In the experimental dental-plaque model, 50% of the PS-negative Strep. mutans were killed by neutrophils while the viability of PS-positive Strep. mutans was not different than of the control. Degranulation of neutrophils was not affected by the presence of extracellular PS of Strep. mutans. Artificial stimulation of neutrophils with phorbol myristate acetate also did not enhance the bactericidal effect of neutrophils on PS-positive Strep. mutans. However, PS-positive Strep. mutans elicited oxygen-reactive products from neutrophils, 2-fold less than with PS-negative Strep. mutans. The results indicate that in situ production of bacterial extracellular polysaccharides might be a major virulence factor of Strep. mutans, enabling PS-positive Strep. mutans in the dental-plaque biofilm to evade killing by human neutrophils.
authors.affiliation: Department of Oral Biology, Faculty of Dental Medicine, Hebrew University, Hadassah, Jerusalem, Israel. dorons@cc.huji.ac.il
authors.forename: D
authors.initials: D
authors.lastname: Steinberg
authors.forename: S
authors.initials: S
authors.lastname: Poran
authors.forename: L
authors.initials: L
authors.lastname: Shapira
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D018441:Biofilms; D002273:Carcinogens; D015550:Cell Degranulation / Q000502:physiology; D003773:Dental Plaque / Q000382:microbiology; D006801:Humans; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D008165:Luminol; D008213:Lymphocyte Activation / Q000187:drug effects; D009113:Muramidase / Q000032:analysis; D009504:Neutrophils / Q000201:enzymology / Q000502:physiology*; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis / Q000494:pharmacology*; D019275:Radiopharmaceuticals; D017382:Reactive Oxygen Species; D013295:Streptococcus mutans / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D013395:Sucrose / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism; D014316:Tritium; D014774:Virulence
chemical_list: D000900:Anti-Bacterial Agents; D002273:Carcinogens; D007202:Indicators and Reagents; D011135:Polysaccharides, Bacterial; D019275:Radiopharmaceuticals; D017382:Reactive Oxygen Species; D014316:Tritium; D013395:Sucrose; D008165:Luminol; D009113:Muramidase; D013755:Tetradecanoylphorbol Acetate; D013936:Thymidine