MONOCLONAL ANTIBODIES RAISED AGAINST SENILE PLAQUES AND NEUROFIBRILLARY TANGLES IN ALZHEIMER'S DISEASE

针对阿尔茨海默病中的老年斑块和神经纤维缠结产生的单克隆抗体

• 批准号: 01480281
• 负责人: FUKATSU Ryo
• 金额: $ 3.52万
• 依托单位: SAPPORO MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01480281/
• 关键词: Alzheimer's disease; monoclonal antibody; amyloid deposition; NFT; amyloid beta protein; apoE; SP-40,40; amyloid associated proteins; アルツハイマ-病; アミロイド; Bー蛋白; serum protein 40ー40; 35kDa蛋白; モノクロ-ナル抗体; βー蛋白; Alzheimer's disease; PHF; Amyloid

Cerebral amyloid deposits of varying morphology, and neurofibrillary tangles (NFT) are major neuropathological characteristics in Alzheimer's disease (AD) affected brains.1) Twelve monoclonal antibodies (mcAb's) against cerebral amyloid were established. The antigens recognized were determined to be Abeta, C3, and C4. we characterized proteinchemically the antigen four mcAb's and two mcAb's recognized to be apoE and SP-40,40. The antigens recognized with Two mcAb's remained unidentified.2) Twenty two mcAb's against PHF were established. The reactive antigens were tau protein, neurofilaments, and ubiquitin. The antigens which the rest of mcAb's e recognized were unidentified.3) Using these mcAb's, immunohistochemical study showed that apoE was found in almost all amyloid depositions studied, and that there was a spatial relation between apoE overproducing astrocytes and plaques. NFT were stained with polyclonal apoE Ab, but not with our mcAb's. Competitive ELISA and Western blot analysis combined with thrombolytic digestion of apoE indicated that our four mcAb's recognized the epitopes within 22 kDa amino-terminal domain of apoE,and there were at least two distinct epitopes of the amino-terminal domain in amyloid depositions. SP-40,40 is present in various morphologies of Abeta depositions to a lessor degree and in astrocytes which are also located close to these depositions in brains affected by AD,and SDAT.These studies suggest that the mechanisms underlying the expression and processing of apoE,and SP-40,40 may be an essential issue in the pathogenesis of AD.

不同形态的脑淀粉样蛋白沉积和神经纤维缠结（NFT）是阿尔茨海默病（AD）影响脑的主要神经病理学特征。1）建立了12种抗脑淀粉样蛋白的单克隆抗体（McAb）。确定识别的抗原为Abeta、C3和C4。我们对抗原进行了蛋白化学表征，四种单克隆抗体和两种单克隆抗体被识别为apoE和SP-40，40。2）建立了22株抗PHF单克隆抗体。反应性抗原为tau蛋白、神经丝和泛素。3）免疫组化结果显示，apoE在所有的淀粉样沉积物中均有表达，且apoE过度表达的星形胶质细胞与斑块之间存在一定的空间关系。NFT用多克隆apoE Ab染色，但不用我们的mAb染色。竞争性ELISA和Western blot结合溶血栓酶切分析表明，4株单抗均识别apoE 22 kDa氨基端结构域内的表位，且在淀粉样蛋白沉积中至少存在2个不同的氨基端结构域表位。SP-40，40以较小程度存在于各种形态的Abeta沉积物中，并且存在于也位于受AD和SDAT影响的脑中的这些沉积物附近的星形胶质细胞中。这些研究表明，apoE和SP-40，40的表达和加工的潜在机制可能是AD发病机制中的重要问题。

项目成果

Takamaru Y et al: "SP-40,40 as an amyloid associated protein in Alzheimer's disease affected brains" (in preparation). (1995)

Takamaru Y 等人：“SP-40,40 作为阿尔茨海默病影响大脑中的淀粉样蛋白相关蛋白”（准备中）。

Aizawa Y,Fukatsu R,Takamaru Y,Tsuzuki K,Obara T,Fujii M,Kobayashi M,Takahata N,Gotoda T,Nagasawa S,Oguma K,Kunishita T and Tabira T: "Monoclonal antibodies against senile plaque amyloid in Alzheimer's disease" In Nagatsu T,Fisher A,Yoshida M (eds). Plenum

Aizawa Y、Fukatsu R、Takamaru Y、Tsuzuki K、Obara T、Fujii M、Kobayashi M、Takahata N、Gotoda T、Nagasawa S、Oguma K、Kunishita T 和 Tabira T：“针对阿尔茨海默病中老年斑淀粉样蛋白的单克隆抗体”

Yuji Aizawa: "Monoclonal antibodies raised against native amyloid in Alzheimer's disease ーestablishment and characterizationー"

Yuji Aizawa：“针对阿尔茨海默病中的天然淀粉样蛋白产生的单克隆抗体 - 建立和表征 -”

Yuji Takamaru: "Serum protein 40ー40 is involved in amyloid plaque formation in Alzheimer's desease."

Yuji Takamaru：“血清蛋白 40-40 参与阿尔茨海默病中淀粉样斑块的形成。”

Tsuzuki K et al: "Potentially amyloidogenic fragment of 50 kDa and intracellular processing of amyloidprecursor protein in cell under leupept in" Brain Research. 659. 213-220 (1994)

Tsuzuki K 等人：“大脑研究中 leupept 下细胞中 50 kDa 的潜在淀粉样蛋白生成片段和淀粉样前体蛋白的细胞内加工”。