ADHERENCE OF PERIODONTAL DISEASE-ASSOCIATED BACTERIA

牙周病相关细菌的粘附

• 批准号: 3219429
• 负责人: WILLIAM B CLARK
• 金额: $ 12.6万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-04-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219429
• 关键词: Actinomyces; adhesin; chromatography; enzyme linked immunosorbent assay; genetic strain; gingival sulcus; glycoproteins; human tissue; immunization; immunochemistry; immunoelectrophoresis; laboratory mouse; laboratory rabbit; microorganism immunology; monoclonal antibody; mutant; oral bacteria; oral mucosa; periodontium disorder; physical chemical interaction; pilus; preventive dentistry; saliva; serum; tissue /cell culture

Periodontal diseases are a major cause of tooth loss in adults. Recognition that microorganisms adhere to tissues such as teeth by specific structures (i.e., adhesins) suggest that regimes might be developed to inhibit adherence and colonization. The mechanisms by which periodontal disease-associated microorganisms attach to an accumulate on the teeth must be understood if periodontal diseases are to be controlled in this manner. The proposed project is a continuation of our studies on the mechanisms of adsorption by Actinomyces viscosus and A. naeslundii to teeth. One facet of the project is directed at characterizing the receptor binding region of the type 1 fimbriae which mediates attachment of A. viscosus to saliva-treated hydroxyapatite (SHA) in vitro. These studies will be approached be immunochemical analysis of isolated fragments of the immunoglobulins using SDS-PAGE gels and Western Blots. By combining the SDS-PAGE, HPLC, and Western Blot techniques with the receptor binding inhibition assay we plan to identify and isolate the receptor binding region type 1 fimbriae which mediates adsorption to SHA. Once this fragment has been identified monospecific and monoclonal antibodies will be prepared against it. We will then examine type 1 fimbriae from other Actinomyces strains to determine whether or not the receptor binding regions of these strains cross-react with that of strain T14V. Further we plan to use fimbrial-deficient mutant strains of strain T14V to determine if the proposed roles for type 1 and type 2 fimbriae in adherence and colonization established in vitro can be confirmed in vivo in humans and mice. Studies demonstrating that the teeth of mice immunized locally in the region of the salivary gland with a mixture of these fimbriae are resistant to colonization when challenged by that bacterial strain, will be expanded. These studies will determine the optimum dose and route of immunization, and whether type 1 or type 2 fimbriae singlely is as effective an immunogen as the mixture. The required role for fimbriae-specific salivary IgA in inhibiting adsorption or reducing colonization will also be evaluated. The principals established for reducing strain T14V colonization of teeth in mice by vaccination with fimbrial adhesins should by helpful in preventing colonization of other periodontopathogens by vaccination with other fimbrial adhesins.

牙周病是成年人牙齿脱落的主要原因。 认识到微生物通过特定的方式附着在牙齿等组织上 结构(即粘附素)表明，系统可能会发展成 抑制附着性和殖民性。牙周的作用机制 附着在牙齿上的与疾病相关的微生物必须 如果要以这种方式控制牙周病，就必须了解。 这项拟议的项目是我们对 粘性放线菌和奈氏放线菌对牙齿的吸附作用一个方面 该项目的主要目的是确定其受体结合区 介导粘性支原体附着的1型菌毛 唾液处理羟基磷灰石(SHA)的体外实验。这些研究将是 应用免疫化学方法对猪瘟病毒分离片段的分析 用SDS-PAGE凝胶和Western blotting检测免疫球蛋白。通过将 结合受体的十二烷基硫酸钠-PAGE、高效液相色谱和蛋白质印迹技术 抑制实验我们计划鉴定和分离受体结合 区域类型1菌毛，介导对SHA的吸附。一旦这一次 片段已被鉴定为单特异性和单抗 做好了应对的准备。然后，我们将检查来自其他地方的1型菌毛 放线菌菌株测定受体是否结合 这些毒株的区域与T14V毒株的区域发生交叉反应。更进一步，我们 计划使用菌株T14V菌毛缺陷突变株来确定 如果类型1和类型2的菌毛在粘连和 在体外建立的定植可以在体内证实在人类和 老鼠。研究表明，在当地免疫的小鼠的牙齿 唾液腺中含有这些菌毛的区域是 当受到该细菌菌株的挑战时，对定植的抗性将是 扩大了。这些研究将确定最佳剂量和途径 免疫，以及1型或2型菌毛是否单独为AS 有效的免疫原作为混合物。所需的角色 菌毛特异性唾液免疫球蛋白A抑制吸附或还原 还将对殖民活动进行评估。为以下目的而建立的原则 减毒株T14V在小鼠牙齿中的定植 菌毛粘附素应有助于防止其他 通过接种其他菌毛粘附素接种牙周病原体。