FIBRONECTIN RECEPTOR MEDIATED ADHESION OF P GINGIVALIS

纤连蛋白受体介导的牙龈卟啉单胞菌粘附

• 批准号: 3425697
• 负责人: KAREN SNAPP
• 金额: $ 2.96万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-01 至 1994-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3425697
• 关键词: Bacteroides gingivalis; antigen receptors; binding proteins; bioassay; cell adhesion; clone cells; enzyme linked immunosorbent assay; fibronectins; flow cytometry; gene expression; human tissue; laboratory rabbit; periodontium disorder; receptor binding; saliva; serum; tissue /cell culture

Approximately 75% of the adult population suffers from some form of periodontal disease. Periodontal disease results when virulent gram negative bacteria colonize the gingival crevice. This colonization involves specific interactions between bacterial adhesions and epithelial cell surface receptors. Secreted molecules found in serum, saliva, and crevicular fluid can also participate in these adhesion events. Porphyromonas gingivalis is a gram negative obligate anaerobe that is commonly isolated from periodontal pockets. Its adherence to oral epithelium is well documented, but the molecular basis for this interaction is unknown. Fibronectin is known to participate in a number of important adhesion events involving the integrin family of receptors, and a number of these receptors have been identified on epithelial cells. In addition, fibronectin can be expressed in an insoluble form on the surface of epithelial cells and also secreted by them. Also, soluble fibronectin is found in both saliva and serum. Finally, several bacterial strains have been identified which are able to bind various forms of human fibronectin. Thus the objective of this proposal is to determine the role of fibronectin in the adhesion of P. gingivalis to oral epithelial cells. Once the role of fibronectin in bacterial adhesion is determined, preventive therapies could to be developed which would inhibit this initial colonization event. Accordingly, the specific aims of this proposal are: (1) to characterize the interactions between P. gingivalis and fibronectin; (2) to characterize the fibronectin expression and/or fibronectin binding capacity of oral epithelium; and (3) to determine the potential role of fibronectin in the binding of P. gingivalis to oral epithelium. The ability of P. gingivalis to interact with either soluble and/or insoluble fibronectin will be determined by means of an ELISA. Both an epithelial cell line and freshly isolated oral epithelial cells will be analyzed by flow cytometry to determine the presence of cell surface fibronectin, receptors for fibronectin, and the ability of the receptors to bind fibronectin. Adhesion assays will be performed to determine the binding properties of P. gingivalis and oral epithelium. Importantly, the ability to use an established cell line as an in vitro model for bacterial adhesion in the oral cavity will also be investigated.

大约 75% 的成年人患有某种形式的疾病 牙周病。 致命的革兰氏菌会导致牙周病 阴性细菌在牙龈缝隙中定殖。 这种殖民 涉及细菌粘附和上皮细胞之间的特定相互作用 细胞表面受体。 血清、唾液和唾液中发现的分泌分子 裂隙液也可以参与这些粘连事件。 牙龈卟啉单胞菌是一种革兰氏阴性专性厌氧菌， 通常从牙周袋中分离出来。 其坚持口头 上皮细胞已被充分记录，但这种相互作用的分子基础 未知。 已知纤连蛋白参与许多重要的 涉及整合素受体家族的粘附事件，以及许多 已在上皮细胞上鉴定出这些受体。 此外， 纤连蛋白可以以不溶性形式表达在细胞表面 上皮细胞并由它们分泌。 此外，可溶性纤连蛋白 唾液和血清中均发现。 最后，一些细菌菌株 已鉴定出能够结合各种形式的人纤连蛋白。 因此，该提案的目的是确定纤连蛋白的作用 牙龈卟啉单胞菌与口腔上皮细胞的粘附。 一旦角色 确定细菌粘附中的纤连蛋白，预防性治疗 可以开发出抑制这种最初的殖民事件的技术。 因此，本提案的具体目标是：（1）表征 牙龈卟啉单胞菌和纤连蛋白之间的相互作用； (2) 表征 口服的纤连蛋白表达和/或纤连蛋白结合能力 上皮； (3) 确定纤连蛋白在 牙龈卟啉单胞菌与口腔上皮的结合。 牙龈卟啉单胞菌与可溶性和/或 不溶性纤连蛋白将通过 ELISA 测定。 两者都是 上皮细胞系和新鲜分离的口腔上皮细胞将被 通过流式细胞术分析以确定细胞表面的存在 纤连蛋白、纤连蛋白受体以及受体的能力 结合纤连蛋白。 将进行粘附力测定以确定 牙龈卟啉单胞菌和口腔上皮的结合特性。 重要的是， 能够使用已建立的细胞系作为细菌的体外模型 还将研究口腔中的粘附。