FIBRONECTIN-BUCCAL EPITHELIAL CELL INTERACTIONS

纤连蛋白-颊上皮细胞相互作用

• 批准号: 3220793
• 负责人: DAVID L HASTY
• 金额: $ 9.99万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1988-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3220793
• 关键词: Escherichia coli; acid base balance; cell adhesion; enzyme linked immunosorbent assay; epithelium; fluorescence microscopy; gel electrophoresis; human tissue; immunoelectrophoresis; ionic strengths; microorganism immunology; monoclonal antibody; oral bacteria; oral mucosa; radioimmunoassay; radiotracer; saliva

The overall goal of this research project is to determine the role of fibronectin (FN) in the attachment of bacteria to the mucosal surface of the oropharynx. To pursue these studies logically, the experiments proposed in this first application are focused on characterizing the interaction of FN with human buccal epithelial cells. Specific efforts will be directed toward: (1) examining buccal cells for content and distribution of FN bound to their surfaces naturally and after experimental treatment, (2) studying the specific characteristics of the binding of exogenous FN to the buccal cells, and (3) determining the major functional domain of plasma FN that mediates binding of the intact molecule to buccal epithelial cells. Pools of buccal epithelial cells will be examined in assays of the amount of labeled anti-FN antibody that binds to the cells, the amount of labeled FN that binds to the cells and the morphological distribution of anti-FN antibodies that bind to the cells. Several different chemical and enzymatic treatments will be used to manipulate FN levels on these cells. Binding studies will be performed to determine the specificity, affinity and number of FN binding sites on buccal epithelial cells. We will pay particular attention to the effects of pH, ionic strength and divalent cation composition and concentration. Well defined FN fragments, and other specific ligands known to interact with FN, will be tested for their ability to inhibit the binding of intact FN to buccal cells. The ability of defined FN fragments to bind to the cells directly will also be tested. Further, defined monoclonal antibodies will be tested for their ability to inhibit the binding of FN to the cells. We believe the proposed experiments on characterization of FN-buccal epithelial cell interactions will be an important step in our efforts to understand the role of FN in bacterial adherence.

该研究项目的总体目标是确定 纤连蛋白 (FN) 促进细菌附着在粘膜表面 口咽部。 为了逻辑地进行这些研究，实验 在第一个应用中提出的重点是表征 FN 与人颊上皮细胞的相互作用。 具体努力 将针对：（1）检查口腔细胞的内容和 自然和实验后结合到其表面的 FN 分布 治疗，（2）研究结合的具体特征 外源性 FN 至颊细胞，以及 (3) 确定主要功能 介导完整分子与颊粘膜结合的血浆 FN 结构域 上皮细胞。 颊上皮细胞池将在 测定与细胞结合的标记抗 FN 抗体的量， 与细胞结合的标记 FN 的量和形态 与细胞结合的抗 FN 抗体的分布。 一些 将使用不同的化学和酶处理来操纵 FN 这些细胞上的水平。 将进行结合研究以确定 颊上皮上 FN 结合位点的特异性、亲和力和数量 细胞。 我们将特别关注 pH 值、离子的影响 强度和二价阳离子组成和浓度。 明确定义 FN 片段以及已知与 FN 相互作用的其他特定配体将被 测试了它们抑制完整 FN 与颊粘膜结合的能力 细胞。 确定的 FN 片段直接与细胞结合的能力 也将受到考验。 此外，还将测试特定的单克隆抗体 因为它们能够抑制 FN 与细胞的结合。 我们相信 拟议的 FN 颊上皮细胞表征实验 互动将是我们努力理解的重要一步 FN 在细菌粘附中的作用。