ROLE OF PEPTIDE CHEMOTACTIC FACTORS IN THE PATHOGENESIS OF RHEUMATOID ARTHRITIS

类风湿关节炎(RA)是一种慢性、进行性、退行性的关节 造成巨大发病率和超额死亡率的疾病。在RA， 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显著的浸润性 血液来源的白细胞。这些细胞介体启动和维持 导致类风湿关节炎关节损伤的炎症反应。惊人的一击 而类风湿关节炎中无法解释的现象是时空上的分离 炎性细胞类型：早期急性皮损以中性粒细胞为主 在滑液中，而单个核细胞在滑膜中积聚 组织。最近发现的两种多肽趋化因子可以解释 中性粒细胞激活因子(NaF)与单核细胞 趋化蛋白-1(MCP)是人类免疫缺陷病毒基因家族中的相关成员。 对其命名的细胞类型具有选择性趋化作用的小肽。 这一提议的基本假设是选择性白细胞 在RA关节中的积累是局部产生NaF和MCP的结果。至 验证这一假设，无论是人类RA滑膜还是 将采用的类风湿关节炎动物模型：兔膝卵白蛋白关节炎模型， 由关节内注射卵白蛋白引起的先前 敏感的动物。 第一个实验目标是定位NaF和MCP蛋白 免疫组织化学和原位杂交检测产物和mRNA 分别在关节炎关节处，并对特定部位进行相关分析 细胞型蓄积，局部分泌NaF和MCP。必需品 先决条件是产生克隆的cdna，重组蛋白， 和抗NaF和MCP的中和抗体在兔身上进行这些研究 和人体组织。这些将被用来确定NaF&的表达 MCP肽和mRNA在关节炎病变演变过程中的表达 在人类类风湿关节炎和兔模型中。在下一阶段的研究中，我们 将确定是否将重组兔NaF和MCP直接注射到 兔膝关节会引起关节炎。此外，我们还将评估 基因转移技术在体内诱导表达的潜力 在滑膜组织中克隆了NaF和MCP基因。最后，我们将评估 氟化钠和单核细胞趋化蛋白在关节炎病变中的作用 抗NaF和MCP的中和性抗体是否抑制血管生成 关节炎的任何方面。对调控机制的理解 RA关节内的炎性细胞可能为 开发新的有效的治疗策略。

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.