IMMUNOCHEMICAL STUDIES OF ACTINOMYCES VISCOSUS POLYSACCHARIDES

粘性放线菌产生的多糖可促进 这些微生物对吸附在牙釉质上或暴露在牙釉质上的糖蛋白的影响 根面 它们可以抑制或增强凝集素介导的共聚集 口腔细菌之间，并有助于大量的牙菌斑。 这些 多糖还可以诱导抗体的形成， 促进龋齿的发展。 虽然A.粘性产生 几种多糖，它们的化学精细结构仍然是 阐明。 它们在根面病变中的作用评价 也没有龋齿。 因此，本提案旨在： 1-在完整的A.粘液细胞定量 荧光染料标记的凝集素结合试验， 结合位点的性质、数量和亲和力（Ka）; 2-以纯化形式分离细胞壁、细胞壁缔合物和 从A. T6、T14 V和T14 Av菌株中提取胞外多糖。 粘度和表征其分子结构的化学和 免疫化学方法; 3-为了检测分离的多糖作为抗原的能力， 抑制细菌附着到唾液涂覆的羟基磷灰石涂覆的 珠粒和抑制细菌共聚集; 4-检测鼠、兔抗体结合位点特异性 多克隆抗体分离A.粘多糖抗原; 5-用分离的多糖抗原主动免疫大鼠， 确定特定聚合物刺激抑制的能力， 口腔感染后疾病加重， 6-制备抗分离多糖的鼠单克隆抗体 抗原，阐明其结合位点的特异性，并评估其作用 的特定抗碳水化合物在修改植入和疾病， 无特定病原体的大鼠。

Polysaccharides produced by Actinomyces viscosus may promote adherence of those microorganisms to glycoproteins adsorbed to enamel or exposed root-surfaces. They may inhibit or enhance lectin mediated coaggregation between oral bacteria, and contribute to the bulk of plaque. These polysaccharides can also induce formation of antibodies that may hinder or promote development of dental caries. Although A. viscosus produces several polysaccharides their chemical fine structure remains to be elucidated. Evaluation of their role in the pathogenesis of root surface caries is also lacking. This proposal therefore aims: 1- on the surface of intact A. viscosus cells by quantitative fluorochrome-tagged lectin binding assays which yield information on the nature, number and affinity (Ka) of binding sites; 2- to isolate in purified form, cell wall, cell wall associated and extracellular polysaccharides from the T6, T14V and T14Av strains of A. viscosus and characterize their molecular structure by chemical and immunochemical methods; 3- to examine the ability of isolated polysaccharides to serve as antigen, inhibit attachment of bacteria to saliva coated hydroxylapatite coated beads and inhibit bacterial coaggregation; 4- to examine the antibody combining site specificity of rat and rabbit polyclonal antibody to isolate A. viscosus polysaccharide antigens; 5- to actively immunize rats with isolated polysaccharide antigens to determine the ability of specific polymers to stimulate suppression or enhancement of disease upon oral infection and 6- to prepare monoclonal rat antibodies against isolated polysaccharide antigens, elucidate their combining site specificity and evaluate the role of specific anti-carbohydrate in modifying implantation and disease in specific pathogen-free rats.