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Immortalization of prion protein deficient cell lines and establishment of prion-susceptible susceptible cell lines by introduction of prion protein genes.

朊病毒蛋白缺陷细胞系的永生化和通过引入朊病毒蛋白基因建立朊病毒敏感的易感细胞系。

基本信息

批准号：
20380166
负责人：
ONODERA Takashi
金额：
$ 12.15万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2010
项目状态：
已结题

项目摘要

After prion infection, an abnormal isoform of prion protein (PrP (Sc)) converts the cellular isoform of prion protein (PrP (C)) into PrP(Sc). PrP (C)-to-PrP (Sc) conversion leads to PrP (Sc) accumulation and PrP (C) deficiency, contributing etiologically to induction of prion diseases. Presently, most of the diagnostic methods for prion diseases are dependent on PrP (Sc) detection. Highly sensitive/accurate specific detection of PrP (Sc) in many different samples is a prerequisite for attempts to develop reliable detection methods. Towards this goal, several methods have recently been developed to facilitate sensitive and precise detection of PrP (Sc), namely, protein misfolding cyclic amplification, conformation-dependent immunoassay, dissociation-enhanced lanthanide fluorescent immunoassay, capillary gel electrophoresis, fluorescence correlation spectroscopy, flow microbead immunoassay, etc. Additionally, functionally relevant prion-susceptible cell culture models that recognize the complexity of the mechanisms of prion infection have also been pursued, not only in relation to diagnosis, but also in relation to prion biology. Prion protein (PrP) gene-deficient neuronal cell lines that can clearly elucidate PrP (C) functions would contribute to understanding of the prion infection mechanism. In this review, we describe the trend in recent development of diagnostic methods and cell culture models for prion diseases and their potential applications in prion biology.

在PrP(SC)感染后，PrP(SC)的异常亚型将细胞内的PrP(C)转变为PrP(SC)。PrP(C)到PrP(SC)的转换导致PrP(SC)的蓄积和PrP(C)的缺乏，在病因学上导致了PrP(SC)疾病的发生。目前，对PrP(Sc)的检测是诊断PrP(Sc)病的主要手段。对许多不同样品中PrP(SC)的高灵敏/准确的特异性检测是尝试开发可靠的检测方法的先决条件。为此，最近发展了几种方法来促进对PrP(SC)的灵敏和精确的检测，即蛋白质错误折叠循环扩增、构象依赖免疫分析、解离增强的稀土元素荧光免疫分析、毛细管凝胶电泳、荧光相关光谱、流动微珠免疫分析等。此外，还寻求了功能相关的、认识到PrP(SC)感染机制复杂性的Prion敏感细胞培养模型，不仅与诊断有关，而且与Prion生物学有关。PrP基因缺失的神经细胞株能够清楚地阐明PrP(C)的功能，有助于了解PrP感染的机制。在这篇综述中，我们描述了最近对病毒病的诊断方法和细胞培养模型的发展趋势以及它们在病毒生物学中的潜在应用。