ADHERENCE OF PERIODONTAL DISEASE-ASSOCIATED BACTERIA

牙周病相关细菌的粘附

• 批准号: 3219430
• 负责人: WILLIAM B CLARK
• 金额: $ 14.88万
• 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 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219430
• 关键词: Actinomyces; adsorption; affinity chromatography; cell adhesion; disease /disorder prevention /control; electrofocusing; enzyme linked immunosorbent assay; genetic regulation; human subject; hydroxyapatites; immunogenetics; immunoglobulins; immunomodulators; inbreeding; laboratory mouse; linkage mapping; monoclonal antibody; oral bacteria; pellicle; periodontium disorder; pilus; proline; serum; tooth enamel; tooth surface

The long-term objectives of this research are to elucidate molecular mechanisms of attachment for important oral bacteria to teeth, and to develop approaches for modulating attachment and colonization of prominent periodontopathogens in the oral cavity. The Specific Aims of the proposed project: (1) identify, isolate and characterize the adhesion associated with Actinomyces type 1 fimbriae which interacts with proline-rich proteins thought to be receptors in the experimental salivary pellicle; (2) establish relevance of the adhesion on attachment to experimental tooth surfaces and natural teeth in vivo; (3) establish functional relevance (i.e., adherence inhibition activity (ALA), and modulation of infection in vivo) of genetically regulated variations in specificity of serum antibodies from inbred mice immunized with A.viscosus T14V, and investigate the possibility that AIA is genetically regulated in humans. Data obtained from this project will hopefully serve as a model for future studies investigating molecular mechanisms of attachment and colonization of other oral microorganisms associated with periodontal diseases in humans. In addition, if AIA has functional relevance regarding actinomyces colonization and is genetically regulated in humans, it is hoped that principles and methods developed to establish this association can be applied to prominent periodontopathogens as well. These studies should lead to development of better approaches for preventing colonization of periodontopathogens by immune modulation. To identify the adhesion, antibody-mediated adsorption inhibition will be studied in an in vitro hydroxyapatite-bacterial adsorption assay; and biochemical (Fast Protein Liquid Chromatography, differential and sucrose- gradient centrifugation, etc.) and immunologic (affinity-chromatography using polyclonal or monoclonal anti-adhesion antibodies) methods will be used for the purification of the adhesion. Molecular mechanisms established in vitro will be confirmed in humans where possible by studying the adsorption of antibiotic-resistant fimbrial-deficient and adherence- defective mutants of A.viscosus T14V to enamel slabs cut from extracted 3rd molars. The functional relevance (i.e., AIA) of genetically regulated variations in the specificity of serum antibodies from inbred mice will be evaluated by testing various spectrotypes and idiotypes of antibodies purified from sera of immunized mice using isoelec-tric focusing and idiotype-specific ELISAs, in AIA assays. Preliminary experiments will also investigate the possibility that AIA is genetically regulated in humans by segregation and linkage analysis of AIA using sera from a group of families who have been previously typed for HLA on chromosome 5 and immunoglobulin allotype on chromosome 11. To establish whether genetic regulation of AIA influences oral colonization, inbred mice which produce high or low levels of AIA will be immunized with purified type 1 fimbriae-associated adhesion and challenged with strain T14V to determine if AIA levels correlate with the level of T14V colonization on teeth.

这项研究的长期目标是阐明分子 重要口腔细菌附着在牙齿上的机制 开发调节突出的依恋和殖民化的方法 口腔中的牙周病原体。 拟议的具体目标 项目：（1）识别、分离和表征相关的粘附 1 型放线菌菌毛与富含脯氨酸的蛋白质相互作用 被认为是实验唾液膜中的受体； (2) 确定实验牙齿附着力的相关性 体内表面和天然牙齿； (3)建立功能相关性 （即粘附抑制活性（ALA）和感染的调节 体内）血清特异性的遗传调节变异 来自用 A.viscosus T14V 免疫的近交小鼠的抗体，并研究 AIA 在人类中受到基因调控的可能性。 获得的数据 该项目有望成为未来研究的模型 研究其他物质的附着和定植的分子机制 与人类牙周病相关的口腔微生物。 在 此外，AIA 是否与放线菌具有功能相关性 定植并在人类中受到基因调控，希望 建立该协会的原则和方法可以 也适用于显着的牙周病原体。 这些研究应该 导致开发出更好的方法来防止殖民化 通过免疫调节抑制牙周病原体。 为了识别粘附，抗体介导的吸附抑制将是 研究了体外羟基磷灰石细菌吸附测定；和 生化（快速蛋白液相色谱、差异和蔗糖- 梯度离心等）和免疫学（亲和层析 使用多克隆或单克隆抗粘附抗体）方法将是 用于纯化粘附物。 分子机制 体外建立的方法将尽可能通过研究在人体中得到证实 抗生素耐药菌毛缺陷的吸附和粘附- A.viscosus T14V 的缺陷突变体对从提取的第三块中切割的牙釉质板 臼齿。 基因调控的功能相关性（即AIA） 近交系小鼠血清抗体特异性的差异将 通过测试抗体的各种光谱型和独特型进行评估 使用等电聚焦从免疫小鼠的血清中纯化 AIA 测定中的独特型特异性 ELISA。 初步实验还将 研究人类 AIA 受基因调控的可能性 使用一组家族的血清进行 AIA 分离和连锁分析 之前已进行过 5 号染色体 HLA 和免疫球蛋白分型的人 11 号染色体上的同种异型。确定 AIA 是否受到遗传调控 影响口腔定植，产生高或低水平的近交小鼠 AIA 将通过纯化的 1 型菌毛相关粘附进行免疫 并用菌株 T14V 进行挑战，以确定 AIA 水平是否与 牙齿上 T14V 定植的水平。