RHEUMATOID ARTHRITIS PHOSPHOLIPASES AND INFLAMMATION

类风湿关节炎磷脂酶和炎症

• 批准号: 3159421
• 负责人: JOHN S BOMALASKI
• 金额: $ 6.56万
• 依托单位: ALLEGHENY UNIVERSITY OF HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3159421
• 关键词: Hymenoptera; arachidonate; binding proteins; cell membrane; eicosanoids; enzyme induction /repression; enzyme linked immunosorbent assay; high performance liquid chromatography; human subject; inflammation; insect poison; laboratory mouse; laboratory rabbit; liposomes; lysosomes; membrane lipids; micelles; monocyte; neutrophil; phospholipids; reptile poison; rheumatoid arthritis; unsaturated fatty acids

Little is known about phospholipase activating proteins in mammals and nothing has previously been described concerning their role in human disease states. The objective of this application is to identify and isolate a phospholipase A2 activating protein from patients with rheumatoid arthritis: to correlate the presence of this activating protein with clinical characteristics of rheumatoid arthritis patients compared to patients with other forms of arthritis; to examine biochemical and cell biological characteristics of these phospholipase A2 activating proteins; and to begin to explore the in vivo relevance of these activators in animal models. Rheumatoid arthritis is an important, common and often disabling disease of joints characterized by an extensive inflammatory reaction. Prostaglandins and related eicosanoids, which are produced in enhanced quantities in this disease, help mediate this destruction. The rate limiting step in the eicosanoid synthesis pathway is phospholipase enzyme cleavage of unsaturated fatty acids such as arachidonic acid from membrane phospholipids. We have previously shown that cells from rheumatoid patients express enhanced phospholipase enzyme activities compared to control cells. Phospholipase activating proteins such as melittin are found in venoms from bees and snakes. Thus, similar proteins might be hypothesized to teleologically exist in humans. Antimelittin antibodies will be used to immuno affinity purify a phospholipase A2 activating protein from rheumatoid tissues, cells and fluids. Further purification will include HPLC gel exclusion and SDS-PAGE. Testing of patient specimens will be performed by immuno dot blot and ELISA assays, as well as histochemical localization. The biochemical characteristics of this protein will include its effects on phospholipase enzymes present in micelles, liposomes, cell membranes and intact cells. The role of parent phospholipid substrate, sn-2 fatty acid substitution, and end product inhibition will be explored. The effects of this protein on lysosomal and cytoplasmic enzyme release from monocytes and neutrophils, and induction of eicosanoid synthesis will be investigated. Preliminary investigations into the in vivo relevance of this protein in animals will include injection of this compound into normal rabbits with quantification of the ensuing inflammatory response, as well as observing the ontologic development of inflammation and the presence of this phospholipase activating protein in animal models of arthritic inflammation.

对磷脂酶激活蛋白知之甚少。 哺乳动物和以前没有描述过的关于 它们在人类疾病状态中的作用。这样做的目的是 应用于鉴定和分离一种激活磷脂酶A2 类风湿关节炎患者的蛋白质：与 这种具有临床特征的激活蛋白的存在 类风湿关节炎患者与其他疾病患者的比较 各种类型的关节炎；检查生化和细胞生物学 这些磷脂酶A2激活蛋白的特性；以及 开始探索这些激活剂在体内的相关性 动物模型。 类风湿性关节炎是一种重要、常见且常致残的疾病 以广泛性炎症为特征的关节病 反应。前列腺素和相关的二十烷类化合物，它们是 在这种疾病中的产量增加，有助于调解这一点 毁灭。二十烷类化合物合成中的限速步骤 不饱和脂肪的磷脂酶分解途径 从膜磷脂中提取的酸，如花生四烯酸。我们 先前已经表明，来自类风湿患者的细胞表达 与对照相比磷脂酶活性增强 细胞。磷脂酶激活蛋白，如蜂毒素 发现于蜜蜂和蛇的毒液中。因此，相似的蛋白质 可能被假设为在目的上存在于人类身上。 抗梅毒蛋白抗体将用于免疫亲和纯化 类风湿组织、细胞磷脂酶A2激活蛋白 还有液体。进一步的纯化将包括高效液相凝胶排除 和十二烷基硫酸钠-PAGE。将对患者样本进行检测 通过免疫斑点印迹和酶联免疫吸附试验以及组织化学 本地化。这种蛋白质的生化特性将 包括其对存在于胶束中磷脂酶的影响， 脂质体、细胞膜和完整的细胞。父母的角色 磷脂底物，sn-2脂肪酸取代，并结束 我们将探讨产物抑制问题。该蛋白对血管生成的影响 单核细胞溶酶体和胞浆酶的释放 中性粒细胞，并诱导二十烷类化合物的合成 调查过了。体内试验的初步研究 这种蛋白质在动物中的相关性将包括注射这个 将复方制成正常兔，并对其进行定量 炎症反应，以及观察本体论 炎症的发展和这种情况的存在 磷脂酶激活蛋白在关节炎动物模型中的作用 发炎。