Hosokawa Tomoko, Ono Fumiko, Tsuchiya Kotaro, Sato Ichiro, Takeyama Natsumi, Ueda Susumu, Zanusso Gianluigi, Takahashi Hidehiro, Sata Tetsutaro, Sakudo Akikazu, Suguira Katsuaki, Baj Andreina, Toniolo Antonio, Yoshikawa Yasuhiro, Onodera Takashi
Nippon Institute for Biological Science, University of Tokyo, Tokyo, Japan.
Microbiol Immunol. 2008 Jan;52(1):25-9. doi: 10.1111/j.1348-0421.2008.00007.x.
By immunizing Prnp-knockout mice with synthetic polypeptides, a panel of mAbs directed to bovine PrP(C) was obtained. The mAb panel was characterized by the ELISA method, where synthetic polypeptides were used for epitope mapping. Different reactivity patterns were identified. The ability of these mAbs to detect abnormal PrP(Sc) in CJD cases was studied by immunohistochemistry. All mAbs were tested for PrP(Sc) in murine, bovine, monkey and human brain tissues. Three mAbs recognized the fragmented PrP epitope in our ELISA. Antibody 1D12 was strongly reactive to ovine and squirrel monkey tissues infected with a scrapie agent, although non-reactive to scrapie-infected mouse tissues. Antibody 2D8 was clearly reactive to type-2 but not type-1 CJD human tissues. Of particular interest was the reactivity of mAb 6C4 with the inner structure of Kuru plaques (peripheral pattern) in a type-2 CJD case and mAb T2, 1D12, 2B11, 2D8, 4B5 and 6G3-2 with the central area (central pattern). The fact that different anti-PrP mAbs possess distinct staining properties suggests that the PrP(c) to PrP(Sc) conversion might involve a multiple-step process.
通过用合成多肽免疫朊蛋白基因敲除小鼠，获得了一组针对牛朊蛋白（PrP）（C型）的单克隆抗体（mAb）。该单克隆抗体组通过酶联免疫吸附测定（ELISA）方法进行表征，其中使用合成多肽进行表位作图。鉴定出了不同的反应模式。通过免疫组织化学研究了这些单克隆抗体检测克雅氏病（CJD）病例中异常朊蛋白（PrP）（Sc型）的能力。所有单克隆抗体均在小鼠、牛、猴和人脑组织中检测PrP（Sc）。在我们的ELISA中，三种单克隆抗体识别了片段化的朊蛋白表位。抗体1D12对感染羊瘙痒病病原体的绵羊和松鼠猴组织有强烈反应，但对感染羊瘙痒病的小鼠组织无反应。抗体2D8对2型克雅氏病人脑组织有明显反应，但对1型无反应。特别有趣的是，在一例2型克雅氏病病例中，单克隆抗体6C4与库鲁病斑块的内部结构（周边模式）有反应，单克隆抗体T2、1D12、2B11、2D8、4B5和6G3-2与中心区域（中心模式）有反应。不同的抗朊蛋白单克隆抗体具有不同的染色特性这一事实表明，朊蛋白（C型）向朊蛋白（Sc型）的转化可能涉及一个多步骤过程。
