FIBRONECTIN-BUCCAL EPITHELIAL CELL INTERACTIONS

这项研究项目的总体目标是确定 纤维连接蛋白(FN)在细菌附着于胃粘膜表面中的作用 口咽部。为了合乎逻辑地进行这些研究，实验 在这第一个应用程序中提出的，重点是描述 纤维连接蛋白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物和 天然和实验后纤维连接蛋白在其表面的分布 治疗，(2)研究结合的特殊性。 外源性FN进入颊上皮细胞，以及(3)测定主要功能 血浆FN结构域，介导完整分子与口腔结合 上皮细胞。口腔上皮细胞池将在 与细胞结合的标记抗FN抗体的量的测定， 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。几个 将使用不同的化学和酶处理来操纵FN 这些细胞上的水平。将进行约束性研究以确定 颊上皮细胞FN结合位点的特异性、亲和力和数量 细胞。我们将特别注意pH、离子的影响 强度和二价阳离子的组成和浓度。定义明确 Fn片段和其他已知与Fn相互作用的特定配体将是 测试它们抑制完整FN与口腔结合的能力 细胞。定义的FN片段直接与细胞结合的能力 也将接受测试。此外，还将测试已定义的单抗。 因为它们能够抑制FN与细胞的结合。我们相信 FN-颊上皮细胞特性的实验研究 互动将是我们努力了解 FN在细菌黏附中的作用。

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.