ROLE OF PEPTIDE CHEMOTACTIC FACTORS IN THE PATHOGENESIS OF RHEUMATOID ARTHRITIS

肽趋化因子在类风湿性关节炎发病机制中的作用

• 批准号: 6100557
• 负责人: RORY M MARKS
• 金额: --
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6100557
• 关键词: chemoattractants; gene expression; histochemistry /cytochemistry; human subject; immunochemistry; in situ hybridization; laboratory mouse; laboratory rabbit; monocyte; neutralizing antibody; neutrophil; nucleic acid probes; pathologic process; polymerase chain reaction; protein biosynthesis; recombinant proteins; rheumatoid arthritis; synovial fluid; transfection

Rheumatoid arthritis (RA) is a chronic, progressive, degenerative joint disease which causes enormous morbidity and excess mortality. In RA, inappropriate activation of inflammatory mechanisms leads to joint injury. Histological assessment of RA synovium reveals prominent infiltrates of blood-derived leukocytes. These cellular mediators initiate and sustain the inflammatory response which leads to joint injury in RA. A striking and unexplained phenomenon in RA is the spatial and temporal disassociation of inflammatory cell types: neutrophils predominate in early acute lesions and in synovial fluid, whereas mononuclear cells accumulate in synovial tissue. Two recently described peptide chemotactic factors could explain this response: Neutrophil Activating Factor (NAF) and Monocyte Chemoattractant Protein-1 (MCP) are related members of a gene family of small peptides which are selectively chemotactic for their named cell type. The underlying hypothesis of this proposal is that selective leukocyte accumulation in RA joints results from local production of NAF and MCP. To test this hypothesis, both human RA synovium and a well-characterized animal model of RA will be used: ovalbumin arthritis in the rabbit knee, elicited by intra-articular administration of ovalbumin to a previously sensitized animal. The first experimental goals will be to localize the NAF and MCP protein products and mRNA by immunohistochemistry and in situ hybridization respectively in the arthritic joint, and to correlate the sites of specific cell-type accumulation with local secretion of NAF and MCP. An essential prerequisite will be to generate the cloned cDNA's, recombinant proteins, and neutralizing antibodies to NAF & MCP to perform these studies in rabbit and human tissue. These will be used to determine the expression of NAF & MCP peptide and mRNA's during the evolution of the arthritic lesion, both in human RA and in the rabbit model. In the next phase of the study, we will determine if direct injection of recombinant rabbit-NAF and MCP into the rabbit knee joint causes arthritis. In addition we will assess the potential of gene transfer techniques to induce in vivo expression of cloned NAF and MCP cDNA's in the synovium. Finally, we will assess the contributions of NAF and MCP to causing the arthritic lesion by determining whether neutralizing antibodies to NAF and MCP inhibit the development of any facet of the arthritis. Understanding of the mechanisms that regulate inflammatory cells within the RA joint could provide the foundation for the development of novel and effective therapeutic strategies.

类风湿性关节炎 (RA) 是一种慢性、进行性、退行性关节病 导致巨大发病率和过高死亡率的疾病。 在RA中， 炎症机制的不当激活会导致关节损伤。 RA 滑膜的组织学评估显示明显的浸润 血液来源的白细胞。 这些细胞介质启动并维持 导致 RA 关节损伤的炎症反应。 一个引人注目的 RA 中无法解释的现象是空间和时间的分离 炎症细胞类型：中性粒细胞在早期急性病变中占主导地位 和在滑液中，而单核细胞在滑液中积累 组织。 最近描述的两种肽趋化因子可以解释 此反应：中性粒细胞激活因子 (NAF) 和单核细胞 趋化蛋白-1 (MCP) 是以下基因家族的相关成员 小肽对其指定的细胞类型具有选择性趋化作用。 该提议的基本假设是选择性白细胞 RA 关节中的积累是 NAF 和 MCP 局部产生的结果。 到 测试这个假设，人类 RA 滑膜和一个充分表征的 将使用RA动物模型：兔膝部卵清蛋白关节炎， 由关节内给予先前的卵清蛋白引起 致敏的动物。 第一个实验目标是定位 NAF 和 MCP 蛋白 通过免疫组织化学和原位杂交检测产物和 mRNA 分别在关节炎关节中，并将特定部位关联起来 局部分泌 NAF 和 MCP 的细胞类型积累。 必不可少的 先决条件是生成克隆的 cDNA、重组蛋白、 以及 NAF 和 MCP 的中和抗体，以在兔子中进行这些研究 和人体组织。 这些将用于确定 NAF & 的表达 MCP 肽和 mRNA 在关节炎病变的演变过程中 在人类 RA 和兔子模型中。 在下一阶段的研究中，我们 将确定重组兔-NAF和MCP是否直接注射到 兔子膝关节会引起关节炎。 此外，我们将评估 基因转移技术诱导体内表达的潜力 在滑膜中克隆了 NAF 和 MCP cDNA。 最后，我们将评估 通过确定 NAF 和 MCP 对引起关节炎病变的贡献 NAF 和 MCP 的中和抗体是否会抑制 关节炎的任何方面。 了解调节机制 RA 关节内的炎症细胞可以为 开发新颖有效的治疗策略。