SELECTINS AND CARBOHYDRATES IN CELL ADHESION AND ARTHRITIS

风湿性关节炎（RA）是一种慢性、进行性、破坏性关节炎 导致巨大发病率和过度死亡率的疾病。 组织学 RA滑膜的评估显示明显的血液来源的 白细胞;这些细胞的激活维持炎症反应 这导致RA中的关节损伤。 炎症性白细胞积聚 病灶是一个多方面的过程，始于细胞粘附， 微血管内皮 这一关键过程源于相互作用 内皮细胞和白细胞表面分子之间，包括 选择素基因家族 我们实验室最近的研究发现 作为碳水化合物的E-选择素的配体， 刘易斯x血型相关分子（sLex）。 该建议的基本假设是：（1）白细胞 RA关节中的积累由选择蛋白的选择性表达诱导， 和（2）抑制选择素-配体结合将抑制白细胞 积累，防止关节损伤。 为了验证这些假设，滑膜 来自RA患者和来自两种良好表征的RA动物模型 （卵清蛋白诱导的兔膝关节炎和链球菌细胞壁 大鼠的关节炎）。 研究计划的一个基本先决条件是产生 克隆的cDNA、重组蛋白和中和抗体， 在兔、大鼠和人体组织中进行研究的选择素。 第一 实验的目标将是准备和验证这些活动 试剂，通过评估体外粘附模型中的选择素表达 使用来自每个测试物种的细胞。 一种充分表征的体外 将使用白细胞粘附于内皮的测定来检测 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素蛋白的表达 在关节炎关节，通过免疫组织化学，并评估 特定白细胞类型积聚部位的对应性 局部表达选择素及其配体。 该项目的最后一部分将决定行政管理是否 可溶形式的选择素配体、中和抗体，或 重组选择素抑制关节炎的发展。 这些研究 将描绘选择素分子的发展的贡献， 关节炎病变，他们可以提供科学依据， 基于这些分子的RA的新的治疗干预， 交互.

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.