RHEUMATOID ARTHRITIS PHOSPHOLIPASES AND INFLAMMATION

对磷脂酶激活蛋白的了解很少， 哺乳动物，以前没有描述过关于 在人类疾病中的作用。 的目的 本发明的应用是鉴定和分离磷脂酶A2激活剂， 类风湿性关节炎患者的蛋白质： 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 关节炎的形式;检查生化和细胞生物学 这些磷脂酶A2活化蛋白的特征;和 开始探索这些激活剂在体内的相关性， 动物模型 风湿性关节炎是一种重要的，常见的，往往致残 以广泛的炎症为特征的关节疾病 反应 前列腺素和相关的类花生酸， 在这种疾病中大量产生， 杀伤性 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 酸如花生四烯酸。 我们 先前的研究表明，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，类似的蛋白质 可能被假设为存在于人类身上。 抗蜂毒肽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 和液体。 进一步纯化将包括HPLC凝胶排阻 SDS-PAGE。 将对患者标本进行检测 通过免疫斑点印迹和ELISA测定，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、细胞膜和完整细胞。 父母的角色 磷脂底物，sn-2脂肪酸取代，和末端 将探索产物抑制。 这种蛋白质对 从单核细胞释放溶酶体和细胞质酶， 中性粒细胞和类花生酸合成的诱导将是 研究了 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 将化合物注射到正常家兔中，并定量随后的 炎症反应，以及观察本体 炎症的发展和这种 关节炎动物模型中磷脂酶激活蛋白 炎症

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.