MECHANISM OF A. NAESLUNDII ADHESION TO BUCCAL EPITHELIUM

本研究项目的总体目标是确定 唾液糖蛋白在口腔细菌附着到 口腔的粘膜表面。 为了进行这些研究 从逻辑上讲，本申请中提出的实验集中在 内氏放线菌粘附细胞的机制 颊上皮细胞，唾液大分子的作用 在介导这些配体-受体相互作用中发挥作用。 朝向 该目标的具体努力将针对：（1）隔离 并表征了吸附到唾液中的唾液成分， A的表面。人唾液中的内氏放线菌 凝集素），并介导和粘附这种细菌的口腔 上皮细胞，（2）研究特异性结合特性， 放线菌结合的唾液大分子与A. 内氏菌和颊上皮细胞表面;（3）检测 颊细胞对凝集素含量和分布的影响 自然地结合在它们的表面上， 治疗，和（4）确定的主要功能域 放线菌凝集素介导的结合完整的 分子到A. naeslundii和颊上皮细胞。 到 实现这些目标，乳糖敏感菌毛介导 附着A。内分泌细胞与颊上皮细胞的结合， 用作细菌-上皮细胞相互作用的参数。 吸附在表面的唾液大分子 曲霉将内氏菌洗脱并纯化至均质。 的 放线菌凝集素将被研究其作用， 介导A. naeslundii至上皮细胞。 将在结合试验中检查放射性标记的凝集素 凝集素对细菌和上皮细胞的作用。 结合研究 以确定特异性、亲和力和 细菌和上皮细胞上凝集素结合位点的数量。 将使用几种不同的化学和酶处理方法 来测量唾液和上皮细胞中的凝集素水平。 的 拟议的研究应提供必要的宝贵信息， 了解唾液成分在细菌粘附中的作用。 它也可能被证明在受体疫苗的开发中有用 以防止易感粘膜表面的细菌定植。

The overall goal of this research project is to determine the role of salivary glycoproteins in the attachment of oral bacteria to the mucosal surfaces of the oral cavity. To pursue these studies logically, the experiments proposed in this application are focused on the mechanisms of adhesion of Actinomyces naeslundii to the buccal epithelial cells, and the role of salivary macromolecules play in mediating these ligand-receptor interactions. Towards that goal specific efforts will be directed toward: (1) isolating and characterizing the salivary components that adsorb to the surface of A. naeslundii from human saliva (actinomyces agglutinin) and mediates and adherence of this bacterial to buccal epithelial cells, (2) studying the specific binding characteristics of the actinomyces-bound salivary macromolecule(s) to cells of A. naeslundii and to surfaces of buccal epithelial cells, (3) examining the buccal cells for content and distribution of the agglutinin bound to their surfaces naturally and after experimental treatment, and (4) determining the major functional domain of actinomyces-agglutinin that mediates the binding of the intact molecule to A. naeslundii and to buccal epithelial cells. To achieve these goals, lactose-sensitive-fimbriae mediated attachment of A. naeslundii to buccal epithelial cells will be employed as a parameter of bacteria-epithelial cell interactions. The salivary macromolecules that were adsorbed to the surfaces of A. naeslundii will be eluted and purified to homogeneity. The actinomyces-agglutinin will be investigated for their role in mediating adherence of A. naeslundii to epithelial cells. Radiolabeled agglutinin will be examined in assays of the binding of the agglutinin to bacteria and epithelial cells. Binding studies will be performed to determine the specificity, affinity and number of agglutinin binding sites on bacteria and epithelial cells. Several different chemical and enzymatic treatments will be used to measure agglutinin levels in saliva and on epithelial cells. The proposed studies should provide valuable information necessary to understand the role of salivary components in bacterial adhesion. It may also prove useful in the development of a receptor-vaccine to prevent bacterial colonization of susceptible mucosal surfaces.