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难治性疾病，可引起人类的传染性痴呆。克雅氏病的病原体尚不清楚，但朊病毒假说已在羊瘙痒病中得到证实。朊病毒的纯化方案从CJD脑和非传染性淀粉样变性中产生淀粉样蛋白。CJD的淀粉样纤维蛋白是由特异性蛋白质（朊病毒蛋白）组成的，它不同于其他淀粉样蛋白。免疫组化显示，抗朊病毒蛋白与库鲁斑呈阳性反应，但与其他淀粉样蛋白沉积物不呈阳性反应。此外，我们还报道了库鲁病斑块的嗜Congophilia通过对CJD试剂的灭活程序进行修改。作为第二步，我们制备了抗人朊病毒、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非常小的库鲁斑在人类和小鼠CJD脑切片。在人类CJD中，几乎所有临床病程长（超过1年）的患者的脑内都有库鲁斑。在小鼠CJD中，库鲁斑由小鼠朊病毒蛋白组成，但不是由人朊病毒蛋白组成。因此，库鲁斑可能是CJD的病理标志之一。CJD的诊断很容易通过使用抗朊蛋白的Western印迹技术来证实。朊病毒蛋白可以在来自CJD感染小鼠的5 mg（湿重）脑组织和25至50 mg脾组织中检测到。为了提高朊蛋白检测的灵敏度，我们准备用我们的抗血清建立朊蛋白的放射免疫分析法。

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.