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The defect of biodefense mechanisms in bovine leukocyte adhesion deficiency syndrome (BLAD) as observed from signal transduction pathway

从信号转导途径观察牛白细胞粘附缺陷综合征（BLAD）生物防御机制的缺陷

基本信息

批准号：
09460133
负责人：
KUWABARA Mikinori
金额：
$ 9.28万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

This project was performed to clarify the signal transduction mechanisms for NADPH oxidase activation and phagocytotis by using normal neutrophils and those with bovine leukocyte adhesion deficiency (BLAD) which was genetic deficient in beta2-integrin CR3 corresponding to the receptor of complement iC3b. Various reagents to inhibit NADPH oxidase-related signal transduction were used for this purpose. We found that inhibitors of protein kinase C (PKC), phosphatidyl inositol 3-kinase (PI 3-kinase) and p38 mitogen-activated protein kinase (p38 MAPK) were dose-dependently inhibited superoxide generation from serum-opsonized zymosan (s-OZ)-stimulated neutrophils from BLAD and normal calves, although stimulation of BLAD neutrophils with s-OZ brought about the lower generation of superoxide than that of normal neutrophils with s-OZ.These results indicated that the lack of beta2-integrin CR3 did not influence signal transduction pathways of NADPH oxidase but reduced superoxide production of NA … More DPH oxidase. This reduced NADPH oxidase activity in BLAD was partially recovered by transfusion of CD 18-positive granulocytes to diseased calf. Furthennore, PI 3-kinase and p38 MAPK but not PKC were shown to be required for phagocytotic activity in normal neutrophils. Concerning the intracellular mechanisms of NADPH oxidase activation, the p47phox, one component of NADPH oxidase, is known to be extensively phosphorylated at serines that are located among its C-terminal region and the phosphorylation is a trigger for the activation of NADPH oxidase. Using site-directed mutagenesis of p47phox and p47phox-deficient B cells from human chronic granulomatous disease (CGD), it was shown that the phosphorylation of serines 303/304, 359/370 and possibly serine 379 must take place in order to activate the oxidase. These results seem to be important in not only understanding the signal transduction mechanism of NADPH oxidase activity but also development for therapy of BLAD and p47phox-deficient CGD by the granulocyte or gene transfusion. Less

本项目旨在通过使用正常中性粒细胞和牛白细胞粘附缺陷（BLAD）（其在对应于补体iC 3b受体的β 2-整合素CR 3中存在遗传缺陷）的中性粒细胞来阐明NADPH氧化酶激活和吞噬作用的信号转导机制。为此目的，使用了各种抑制NADPH氧化酶相关信号传导的试剂。我们发现，蛋白激酶C（PKC）、磷脂酰肌醇3-激酶（PI 3-激酶）和p38丝裂原活化蛋白激酶（p38 MAPK）的剂量依赖性抑制血清调理酵母聚糖（s-OZ）刺激的BLAD和正常小牛的中性粒细胞产生超氧化物，虽然用s-OZ刺激BLAD中性粒细胞比用s-OZ刺激正常中性粒细胞产生更低的超氧化物，OZ。这些结果表明β 2-整联蛋白CR 3的缺乏不影响NADPH氧化酶的信号传导途径，但减少了NA的超氧化物产生 ...更多信息 DPH氧化酶。将CD 18阳性粒细胞输注给病牛后，BLAD中NADPH氧化酶活性部分恢复。此外，PI 3-激酶和p38 MAPK而不是PKC被证明是正常中性粒细胞吞噬活性所必需的。关于NADPH氧化酶激活的细胞内机制，已知NADPH氧化酶的一种组分p47 phox在位于其C-末端区域之间的丝氨酸处被广泛磷酸化，并且磷酸化是NADPH氧化酶激活的触发器。使用来自人慢性肉芽肿病（CGD）的p47 phox和p47 phox缺陷型B细胞的定点诱变，显示丝氨酸303/304、359/370和可能的丝氨酸379的磷酸化必须发生以激活氧化酶。这些结果不仅对了解NADPH氧化酶活性的信号转导机制，而且对通过粒细胞或基因输注治疗BLAD和p47 phox缺陷型CGD的发展具有重要意义。少