SELECTINS AND CARBOHYDRATES IN CELL ADHESION AND ARTHRITIS

选择素和碳水化合物在细胞粘附和关节炎中的作用

• 批准号: 6099601
• 负责人: RORY M MARKS
• 金额: $ 11.52万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6099601
• 关键词: antiinflammatory agents; cell adhesion; disease /disorder model; inhibitor /antagonist; laboratory rabbit; ligands; oligosaccharides; receptor; rheumatoid arthritis; selectins; tissue /cell culture; vascular endothelium

Rheumatoid arthritis (RA) is a chronic, progressive, destructive joint disease which causes immense morbidity and excess mortality. Histological assessment of RA synovium reveals prominent infiltrates of blood-derived leukocytes; activation of these cells sustains the inflammatory response which leads to joint injury in RA. Leukocyte accumulation in inflammatory foci is a multi-faceted process that begins with cell adhesion to microvascular endothelium. This crucial process results from interactions between endothelial and leukocyte surface molecules, including members of the Selectin gene family. Recent work from our laboratory has identified the ligand for E-Selectin as a carbohydrate, the sialylated form of the Lewis x blood group related molecule (sLex). The underlying hypotheses of this proposal are that: (1) leukocyte accumulation in RA joints is induced by selective expression of Selectins, and (2) inhibition of Selectin-ligand binding will suppress leukocyte accumulation and prevent joint damage. To test these hypotheses, synovium from patients with RA and from two well-characterized animal models of RA (ovalbumin-induced arthritis in the rabbit knee and streptococcal cell wall arthritis in the rat) will be used. An essential prerequisite for the research program will be to generate cloned cDNA's, recombinant proteins, and neutralizing antibodies to Selectins to perform studies in rabbit, rat, and human tissue. The first experimental goal will be to prepare and validate the activity of these reagents, by assessing Selectin expression in in vitro adhesion models using cells derived from each test species. A well-characterized in vitro assay of leukocyte adhesion to endothelium will be used to test the inhibitory activity of the neutralizing antibodies, sLex and other carbohydrate analogs, and the recombinant Selectin molecules. The next experimental goal will be to localize Selectin protein expression in the arthritic joint, by immunohistochemistry, and to assess the correspondence of the sites of specific leukocyte cell-type accumulation with local expression of Selectins and their ligands. The final component of this project will determine whether administration of soluble forms of Selectin ligands, neutralizing antibodies, or recombinant Selectins inhibits the development of arthritis. These studies will delineate the contribution of Selectin molecules to the development of the arthritic lesion, and they may provide the scientific foundation for novel therapeutic interventions for RA, based on these molecular interactions.

类风湿性关节炎 (RA) 是一种慢性、进行性、破坏性关节 导致巨大发病率和过高死亡率的疾病。 组织学 RA 滑膜评估显示血源性物质明显浸润 白细胞；这些细胞的激活维持炎症反应 这会导致 RA 中的关节损伤。 炎症时白细胞积聚 焦点是一个多方面的过程，从细胞粘附开始 微血管内皮。 这个关键过程是相互作用的结果 内皮细胞和白细胞表面分子之间，包括成员 选择素基因家族。 我们实验室最近的工作已经确定 E-选择素的配体作为碳水化合物，其唾液酸化形式 Lewis x 血型相关分子（sLex）。 该提案的基本假设是：（1）白细胞 选择素的选择性表达会诱导 RA 关节中的积累， (2) 抑制选择素-配体结合将抑制白细胞 积累并防止关节损伤。 为了检验这些假设，滑膜 来自 RA 患者和两种特征良好的 RA 动物模型 （卵白蛋白诱导的兔膝关节炎和链球菌细胞壁 大鼠关节炎）将被使用。 研究计划的一个重要先决条件是产生 克隆的 cDNA、重组蛋白和中和抗体 选择蛋白在兔、大鼠和人体组织中进行研究。 第一个 实验目标是准备和验证这些的活性 试剂，通过评估体外粘附模型中的选择蛋白表达 使用来自每个测试物种的细胞。 具有良好的体外表征 白细胞与内皮细胞粘附的测定将用于测试 中和抗体、sLex 和其他抗体的抑制活性 碳水化合物类似物和重组选择素分子。 下一个实验目标是定位选择蛋白表达 在关节炎关节中，通过免疫组织化学，并评估 特定白细胞细胞类型积累位点的对应关系 选择蛋白及其配体的局部表达。 该项目的最后一部分将决定管理是否 可溶形式的选择蛋白配体、中和抗体，或 重组选择素抑制关节炎的发展。 这些研究 将描述选择素分子对发展的贡献 关节炎病变，它们可能提供科学基础 基于这些分子的新的 RA 治疗干预措施 互动。