Immunological detection of specific protein from organs and blood of patients with Creutzfeldt-Jakob disease.

克雅氏病患者器官和血液中特异性蛋白的免疫学检测。

基本信息

批准号：
61480202
负责人：
TATEISHI Jun
金额：
$ 4.35万
依托单位：
Kyushu 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：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61480202/
关键词：
Creutzfeldt-Jakob disease Pathogen Prion Amyloid クル班脾臓アミロイド斑

项目摘要

Creutzfeldt-Jakob disease (CJD) is a intractable disease of central nervous system, causing transmissible dementia in human. The pathogen of CJD remains unknown, however, the prion hypothesis have been declared in scrapie infections.I investigated the prion hypothesis in CJD. The purification protocols of prion yield amyloid proteins from CJD brains and non-transmissible amyloidosis. The amyloid fibril protein of CJD is composed of specific protein (prion protein), which is different from other amyloid proteins. Immunohistochemically, anti-prion protein reacted positively with kuru plaques in CJD brains, but not with other amyloid deposits. In addition, we reported that Congophilia of kuru plaque is modified by inactivation procedures on CJD agents. As the second step, we produced the antisera against human prion, mouse prion, and prion protein synthetic peptides. These antisera and a newly developed formic acid pretreatment revealed the very small kuru plaques in human and mouse CJD brain sections. In human CJD, almost all patients with long clinical couse (over 1 year) have kuru plaques in their brains. In mouse CJD, the kuru plaques are composed of mouse prion proteins, but not of human prion proteins. Therefore, kuru plaques can be one of the pathological hallmarks in CJD. The diagnosis of CJD is easily confirmed by the Western blotting technique using antiprion proteins. Prion protein can be detected in 5 mg (wet weight) of brain tissues, and 25 to 50 mg of spleen tissues from CJD-infected mice. In order to increase the sensitivity of the prion protein detection, we are going to establish the radioimmunoassay using our antisera.

克雅氏病（CJD）是一种中枢神经系统的难治性疾病，可导致人类传染性痴呆。克雅氏病的病原体尚不清楚，但朊病毒假说已在瘙痒病感染中被宣布。我研究了克雅氏病的朊病毒假说。朊病毒的纯化方案可从克雅氏病大脑和非传染性淀粉样变性中产生淀粉样蛋白。克雅氏病的淀粉样原纤维蛋白由特定蛋白（朊病毒蛋白）组成，与其他淀粉样蛋白不同。免疫组织化学显示，抗朊病毒蛋白与克雅氏病大脑中的库鲁斑块呈阳性反应，但与其他淀粉样蛋白沉积物不呈阳性反应。此外，我们报道了库鲁菌斑的嗜刚果癖通过克雅氏病药物的灭活程序得到了改善。第二步，我们生产了针对人朊病毒、小鼠朊病毒和朊病毒蛋白合成肽的抗血清。这些抗血清和新开发的甲酸预处理揭示了人和小鼠克雅氏病脑切片中非常小的库鲁斑块。在人类克雅氏病中，几乎所有临床病程较长（超过一年）的患者大脑中都有库鲁斑块。在小鼠克雅氏病中，库鲁斑块由小鼠朊病毒蛋白组成，但不是由人类朊病毒蛋白组成。因此，库鲁斑可能是克雅氏病的病理标志之一。使用抗朊病毒蛋白的蛋白质印迹技术可以轻松确诊克雅氏病。在克雅氏病感染小鼠的5毫克（湿重）脑组织和25至50毫克脾组织中可以检测到朊病毒蛋白。为了提高朊病毒蛋白检测的灵敏度，我们将建立使用我们的抗血清的放射免疫分析方法。