ACTIVATION OF THE HUMAN NEUTROPHIL

超氧阴离子的产生和脱粒是关键的， 嗜中性粒细胞的杀微生物功能，但也可能有助于组织 炎症时的损伤。 因此对功能的理解。 这些研究的目的是调查人类的激活 中性粒细胞的吞噬和非吞噬刺激，特别是 参考钙运动、膜脂质和蛋白质的作用 激酶作为刺激-反应偶联的转换器或调节器。 钙和蛋白激酶C与触发和 下调中性粒细胞功能。 配体如趋化性 肽f-Met-Leu-Phe激活磷脂酶C， 细胞内钙，和二甘醇甘油，一种激活 蛋白激酶C将评价甘油二酯的其他来源，包括 磷脂酰肌醇、糖基磷脂酰肌醇、磷脂酰胆碱 以及甘油二酯的从头定位和分子种类， 确定辅因子活性的关键考虑因素。 二酯 是中性粒细胞活化及其清除中的重要信号元件 可以作为细胞活化的重要控制。 定位和 对调节二酰基甘油至关重要的酶的活性，例如 作为二酰基甘油激酶，将被确定。 蛋白激酶C的多种免疫反应性物质，α，β和 γ，N-同工酶，已经在中性粒细胞中得到证实。 是 假设蛋白激酶C的不同同种型发挥选择性作用， 在中性粒细胞功能中的作用 β蛋白激酶C， 从胞质溶胶转移到膜响应于升高的Ca或 佛波醇肉豆蔻酸酯乙酸酯，以及一种新的非易位的α- 蛋白激酶C将被纯化。 差分余因子要求 以及这些不同蛋白激酶C同种型的底物特异性 将被确定为理解阿尔法选择性作用的关键- PKC和β PKC在中性粒细胞功能中的作用。 一种新的糖基-磷脂酰肌醇糖基-PI）已被证明 在中性粒细胞中，它可能通过磷脂酶C发挥骨架作用，并选择性地 抑制超氧化物但不抑制脱粒。 因此， 糖基-P1是用于检查信号传导的独特的生理探针， 产生超氧化物。 对积极的产生的理解 和负面信号，以及它们的调节在设计 为加强东道国所需的这些职能而采取的适当战略 防御和选择性下调负责组织的功能 损害

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.