RHEUMATOID ARTHRITIS PHOSPHOLIPASES AND INFLAMMATION

类风湿关节炎磷脂酶和炎症

• 批准号: 3159426
• 负责人: JOHN S BOMALASKI
• 金额: $ 6.06万
• 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/3159426
• 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.

对于磷脂酶激活蛋白知之甚少 哺乳动物，之前没有任何关于 它们在人类疾病状态中的作用。 此举的目的 应用是鉴定和分离磷脂酶 A2 激活 类风湿性关节炎患者的蛋白质：将 这种激活蛋白的存在具有以下临床特征 类风湿性关节炎患者与其他类风湿性关节炎患者相比 关节炎的形式；检查生化和细胞生物学 这些磷脂酶 A2 激活蛋白的特性；和 开始探索这些激活剂的体内相关性 动物模型。 类风湿性关节炎是一种重要、常见且经常致残的疾病 以广泛炎症为特征的关节疾病 反应。 前列腺素和相关的类二十烷酸，它们是 在这种疾病中产生的数量增加，有助于调节这种情况 破坏。 类二十烷酸合成中的限速步骤 途径是磷脂酶裂解不饱和脂肪 酸，例如来自膜磷脂的花生四烯酸。 我们 之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肽 在蜜蜂和蛇的毒液中发现。 因此，相似的蛋白质 可能被假设为目的论地存在于人类中。 抗利丁抗体将用于免疫亲和纯化 来自类风湿组织、细胞的磷脂酶 A2 激活蛋白 和液体。 进一步纯化将包括 HPLC 凝胶排除 和SDS-PAGE。 将对患者样本进行测试 通过免疫斑点印迹和 ELISA 测定以及组织化学 本土化。 该蛋白质的生化特性将 包括其对胶束中存在的磷脂酶的影响， 脂质体、细胞膜和完整细胞。 家长的角色 磷脂底物、sn-2 脂肪酸取代和末端 将探讨产物抑制。 这种蛋白质的作用 单核细胞释放溶酶体和细胞质酶 中性粒细胞，并诱导类二十烷酸合成 调查了。 体内初步研究 这种蛋白质在动物中的相关性将包括注射这种蛋白质 将化合物注入正常兔子体内，并对随后的结果进行定量 炎症反应，以及观察本体论 炎症的发展及其存在 关节炎动物模型中的磷脂酶激活蛋白 炎。