ADHERENCE OF PERIODONTAL DISEASE-ASSOCIATED BACTERIA

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，以及 开发调节突出的依恋和定植的方法 口腔中的牙周病原体。建议的具体目标 项目：(1)确定、分离和表征相关的粘连 与与富含脯氨酸的蛋白质相互作用的1型放线菌菌毛 被认为是实验性唾液膜的受体；(2) 建立附着体上的粘附力与实验牙的相关性 活体表面和天然牙；(3)建立功能相关性 (即黏附抑制活性(ALA)和对感染的调节 体内)的血清特异性受基因调控的变异 粘性弧菌T14V免疫近交系小鼠的抗体研究 AIA在人类中受到基因调控的可能性。获得的数据 这一项目将有望成为未来研究的典范 探讨其他细菌附着和定植的分子机制 与人类牙周病有关的口腔微生物。在……里面 此外，如果AIA与放线菌有功能相关性 并在人类中受到基因调控，人们希望 为建立这种联系而开发的原则和方法可以是 也适用于突出的牙周病原体。这些研究应该 导致开发更好的方法来防止殖民 牙周病病原体通过免疫调节。 为了鉴定黏附，抗体介导的吸附抑制将是 在体外羟基磷灰石-细菌吸附实验中进行研究； 生物化学(快速蛋白质液相色谱、差示和蔗糖- 梯度离心法等)和免疫学(亲和层析 使用多克隆或单克隆抗黏附抗体)方法将 用于胶粘剂的净化。分子机制 在体外建立的研究将在可能的情况下在人类身上得到确认 抗生素耐药菌毛缺乏的吸附和黏附- 粘性曲霉T14V缺陷突变株对第3代釉质切片的致病作用 磨牙。基因管制的功能相关性(即，AIA) 近交系小鼠血清抗体的特异性将发生变化 通过检测抗体的不同谱型和独特型进行评估 等电聚焦从免疫小鼠血清中提纯 AIA检测中的独特型特异性ELISA。初步实验也将 调查AIA在人类中受基因调控的可能性 利用一组家系血清进行AIA的分离和连锁分析 之前已经对5号染色体上的人类白细胞抗原和免疫球蛋白进行了分型 11号染色体上的同种异型。为了确定AIA的遗传调控 影响口腔定植，近交系小鼠产生高或低水平 将用纯化的1型菌毛相关黏附免疫 并用菌株T14V进行攻击，以确定AIA水平是否与 T14V在牙齿上的定植水平。

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.