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

克雅氏病（Creutzfeldt-Jakob disease， CJD）是一种中枢神经系统的顽固性疾病，是人类的传染性痴呆。克雅氏病的病原尚不清楚，然而，在痒病感染中已经宣布了朊病毒假说。我调查了克雅氏病的朊病毒假说。从CJD脑和非传染性淀粉样变性中提取朊病毒的纯化方案产生淀粉样蛋白。CJD的淀粉样蛋白纤维蛋白由特异性蛋白（朊蛋白）组成，与其他淀粉样蛋白不同。免疫组织化学结果显示，抗朊病毒蛋白与CJD脑库鲁斑块反应阳性，但与其他淀粉样蛋白沉积反应阴性。此外，我们报道了库鲁菌菌斑的嗜血友病通过克雅氏病药物的失活程序进行修饰。第二步，制备了人朊病毒、小鼠朊病毒的抗血清和朊病毒蛋白合成肽。这些抗血清和新开发的甲酸预处理揭示了人类和小鼠CJD脑切片中非常小的库鲁斑块。在人类CJD中，几乎所有临床病程长（超过1年）的患者大脑中都有库鲁菌斑块。在小鼠CJD中，库鲁斑块由小鼠朊病毒蛋白组成，而不是由人类朊病毒蛋白组成。因此，库鲁斑块可能是CJD的病理标志之一。CJD的诊断很容易通过使用抗朊病毒蛋白的Western blotting技术得到证实。在cjd感染小鼠脑组织（湿重）5 mg和脾脏组织（湿重）25 ~ 50 mg中检测到朊病毒蛋白。为了提高朊病毒蛋白检测的灵敏度，我们将利用我们的抗血清建立放射免疫分析法。

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.