Activation of the human neurtrophil

描述(由申请人提供)：由 中性粒细胞对于杀菌活动是必不可少的，但也可以引起 脓毒症、类风湿等疾病中炎症的组织损伤 关节炎、哮喘。肿瘤坏死因子(TNFa)等免疫配体 复合体和趋化肽，如fMet-Leu-Phe，诱导分泌 氧自由基和颗粒含量以及对表面的粘附性。我们假设 离散蛋白激酶C(PKC)亚型既有阳性也有阴性 在促炎反应信号中的作用。促炎信号可以 在多个水平上受PKC亚型的调节，包括：a)受体，b) 产生和调节i)第二信使Ca~(2+)，以及ii)脂质辅助因子， 以及c)组装用于产生O2-的络合物，如NADPH氧化酶。vbl.使用 去除分化HL60中特定PKC亚型的反义策略 (DHL60)，我们展示了a-PKC和b-PKC在 产氧量。相反，b-PKC下调配体启动的钙摄取， 5-PKC对p60TNFR功能具有负性调节作用。 反义策略将被用来选择性地耗尽特定的PKC亚型， 建立脂质和脂质依赖的PKC亚型在信号中的作用 促炎症反应的转导。我们将：我将建立选择性的 钙依赖的a-PKC和b-PKC亚型在NADPH激活中的作用 氧化酶和p47Phox的磷酸化。II确定以下人员的选择角色 钙依赖的a-PKC和b-PKC亚型在黏附和活化中的作用 Marcks脱颗粒和磷酸化。确定PKC亚型 负责调节配体启动的细胞内动员 Ca~(2+)和存储操作的钙通道。IV审查选定的角色 Ca~(2+)依赖的a-PKC和b-PKC亚型对配体启动的调节 涉及效应酶PLCB/PLCg和PLD1及其涉及的酶的信号转导 在PIP2的再生中。确立d-PKC在下调细胞周期中的作用 由p60TNFR触发的信号通路。确定d-PKC是否作用于 丝氨酸残基(S)在膜旁区域或死亡区域。

DESCRIPTION (provided by applicant): Secretion of oxygen radicals by neutrophils is essential for microbicidal activity, but can also give rise to the tissue damage of inflammation in diseases such as sepsis, rheumatoid arthritis, asthma. Ligands such as tumor necrosis factor (TNFa), immune complexes and chemotactic peptides such as fMet-Leu-Phe, elicit secretion of oxygen radicals and granule contents and adherence to a surface. We hypothesize that discrete protein kinase C (PKC) isotypes play both positive and negative roles in signaling for proinflammatory responses. Proinflammatory signaling can be regulated by PKC isotypes at numerous levels including: a) the receptor, b) generation and regulation of i) second messenger Ca2+, and ii) lipid cofactors, and c) assembly of complexes such as the NADPH oxidase for O2-generation. Using antisense strategies to deplete specific PKC isotypes in differentiated HL60 (dHL60), we demonstrated positive signaling roles for both a-PKC and b-PKC for O2- generation. Conversely, b-PKC downregulates ligand-initiated Ca2+ uptake, and 5-PKC negatively regulates the p60TNF receptor (p60TNFR) function. Antisense strategies will be used to selectively deplete specific PKC isotypes, to establish roles for lipids and lipid-dependent PKC isotypes in signal transduction for proinflammatory responses. We will: I Establish selective roles for Ca2+-dependent a-PKC and b-PKC isotypes in activation of the NADPH oxidase, and phosphorylation of p47phox. II Determine selective roles for Ca2+-dependent a-PKC and b-PKC isotypes in activation of adherence and degranulation and phosphorylationof MARCKS. III Determine the PKC isotypes responsible for regulation of ligand-initiated mobilization of intracellular Ca2+ and store operated Ca2+ channels. IV Examine selective roles for Ca2+-dependent a-PKC and b-PKC isotypes in the regulation of ligand initiated signalling involving effector enzymes PLCb/PLCg, and PLD1, and enzymes involved in the regeneration of PIP2. V Establish the role of d-PKC in downregulation of signalling pathways triggered by the p60TNFR. Determine if d-PKC acts on a serine residue(s) in the juxtamembrane region or the death domain.