Takamaru Y
Department of Psychiatry and Neurology, Hokkaido University School of Medicine, Sapporo, Japan.
Hokkaido Igaku Zasshi. 1994 Mar;69(2):191-201.
Amyloid deposition is the major hallmark of Alzheimer's disease (AD). Beta/A4 protein is identified as the major component of amyloid in the brains of AD. But all the components of amyloid and the precise mechanism of amyloid formation are still unknown. To provide better understanding of amyloidogenesis in AD brain, we have established 12 monoclonal antibodies (mcAb's) which immunohistochemically reacted with amyloid or amyloid-related structures, and tried to characterize these mcAb's. The epitopes, which these mcAb's reacted with, have been studied, and 2 of these mcAb's (Az 172/4, Az 520/4) turned out to react with protein of 80 kDa in brain homogenate or normal human plasma (NHP). Reactive antigen was purified from NHP using PEG fractionation, and immunoaffinity column. Amino-terminus amino acid sequence analysis of purified antigen was performed, and the results showed that 80 kDa antigen was identical to SP-40, 40 (or clusterin). Immunohistochemical study revealed that various types of senile plaque and vascular amyloid were stained with the mcAb's. In addition, fibrous background structures were also stained in AD brain. But there were no positive structures in control brains. The above observations indicate that SP-40, 40 is expressed and deposited in AD brain. This protein is closely associated with amyloid deposition. SP-40, 40 seems to play an important role in pathogenesis of amyloid formation in AD.
淀粉样蛋白沉积是阿尔茨海默病(AD)的主要标志。β/A4蛋白被确定为AD患者大脑中淀粉样蛋白的主要成分。但淀粉样蛋白的所有成分以及淀粉样蛋白形成的确切机制仍不清楚。为了更好地理解AD大脑中的淀粉样蛋白生成过程,我们制备了12种单克隆抗体(mcAb),这些抗体与淀粉样蛋白或淀粉样蛋白相关结构发生免疫组织化学反应,并试图对这些mcAb进行特性分析。已经研究了这些mcAb所反应的表位,结果发现其中2种mcAb(Az 172/4、Az 520/4)能与脑匀浆或正常人血浆(NHP)中的80 kDa蛋白发生反应。使用聚乙二醇分级分离和免疫亲和柱从NHP中纯化反应性抗原。对纯化抗原进行了氨基末端氨基酸序列分析,结果表明80 kDa抗原与SP-40、40(或簇集蛋白)相同。免疫组织化学研究显示,各种类型的老年斑和血管淀粉样蛋白都被这些mcAb染色。此外,AD大脑中的纤维状背景结构也被染色。但对照大脑中没有阳性结构。上述观察结果表明,SP-40、40在AD大脑中表达并沉积。这种蛋白与淀粉样蛋白沉积密切相关。SP-40、40似乎在AD淀粉样蛋白形成的发病机制中起重要作用。