ACTIVATION OF THE HUMAN NEUTROPHIL

超氧阴离子的产生和脱颗粒是关键 中性粒细胞的杀菌功能，但也可能对组织有贡献 炎症过程中的损伤。因此对其功能有一定的了解。 这些研究旨在调查人类的激活 吞噬和非吞噬刺激的中性粒细胞，特别是 参考钙运动、膜脂和蛋白质的作用 作为刺激-反应耦合的传送器或调节器的激活酶。 钙和蛋白激酶C都与触发和 下调中性粒细胞功能。趋化配体等 多肽f-Met-Leu-Phe激活磷脂酶C，该酶能裂解 细胞内钙和二氯甘油，一个激活的辅因子 蛋白激酶C。二甘油三酯的其他来源将得到评估，包括 磷脂酰肌醇、糖基磷脂酰肌醇、磷脂酰胆碱 和双甘油酯的从头定位和分子物种，两者都 确定辅因子活性的关键考虑因素。甘油二酯 是中性粒细胞激活和清除的重要信号元件 可能是细胞激活的重要控制因素。本地化和 调节甘油二酯的关键酶的活性，如 作为二酰基甘油激活剂，将被测定。 多种蛋白激酶C、α、β和 在中性粒细胞中已经证实了伽马，n-同工酶。它是 假设不同亚型的蛋白激酶C起选择性作用 在中性粒细胞功能中的作用。β蛋白激酶C，也就是 从细胞质转移到细胞膜以响应升高的钙或 佛波醇肉豆蔻酸酯，以及一种新的非移位α- 蛋白激酶C将被提纯。差异余因数要求 以及这些不同类型的蛋白激酶C的底物特异性 将被确定为理解阿尔法的选择性作用的关键- 中性粒细胞功能中的PKC和βPKC。 证明了一种新的糖基磷脂酰肌醇(糖基-Pl) 在中性粒细胞中，它可能通过磷脂酶C发挥骨干作用，并选择性地 抑制超氧化物，但不能脱颗粒。因此，极头基团 Glycoyl-Pl是一种独特的检测信号转导的生理学探针 用于产生超氧化物。对积极生成的一种理解 和负面信号，以及它们的调节在设计中是必不可少的 增强主机所需功能的适当策略 防御和选择性下调对组织负责的功能 损坏。

Generation of superoxide anion and degranulation are critical to the microbicidal function of neutrophils, but may also contribute to tissue damage during inflammation. Therefore an understanding of the function. These studies are designed to investigate the activation of human neutrophils by phagocytic and non-phagocytic stimuli, with particular reference to the role of calcium movements, membrane lipids, and protein kinases as transducers or modulators of stimulus-response coupling. Calcium and protein kinase C have been implicated in both triggering and down regulating neutrophil functions. Ligands such as the chemotactic peptide f-Met-Leu-Phe, activate a phospholipase C which cleaves intracellular Ca, and diayclglycerol, a cofactor for the activation of protein kinase C. Other sources of diglyceride will be evaluated including phosphatidyl inositol, glycosyl-phosphatidyl inositol, phosphatidyl choline and de novo localization and molecular species of the diglyceride, both critical considerations in determining cofactor activity. Diacylglycerol is an important signalling element in neutrophil activation and its removal may serve as an important control for cell activation. Localization and activity of the enzymes critical to the regulation of diacyl glycerol, such as diacylglycerol kinase, will be determined. Multiple immunoreactive species of protein kinase C, the alpha-, beta and gamma,n-isozymes, have been demonstrated in the neutrophil. It is hypothesized that different isotypes of protein kinase C play selective roles in neutrophil functions. The beta-protein kinase C, that is translocated from cytosol to membrane in response to elevated Ca or to phorbol myristate acetate, as well as a novel non-translocated alpha- protein kinase C will be purified. The differential cofactor requirements and substrate specificities of these different isotypes of protein kinase C will be determined as a key to understanding the selective roles of alpha- PKC and beta PKC in neutrophil functions. A novel glycosyl-phosphatidyl inositol glycosyl-Pl) has been demonstrated in neutrophils, which may play backbone by phospholipase C, and selectively inhibits superoxide but not degranulation. Thus the polar head group of glycosyl-Pl is a unique and physiologic probe for examining the signalling for superoxide generation. An understanding of the generation of positive and negative signals, and of their regulation is essential in devising appropriate strategies for enhancing those functions necessary for host defense and selectively down regulating functions responsible for tissue damage.