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.

朊病毒感染后，异常的朊病毒蛋白异构体 (PrP (Sc)) 将细胞内的朊病毒蛋白异构体 (PrP (C)) 转化为 PrP(Sc)。 PrP (C) 向 PrP (Sc) 的转化导致 PrP (Sc) 积累和 PrP (C) 缺乏，从病因学角度讲，有助于诱发朊病毒疾病。目前，大多数朊病毒病的诊断方法都依赖于PrP（Sc）检测。在许多不同的样品中对 PrP (Sc) 进行高灵敏度/准确的特异性检测是尝试开发可靠检测方法的先决条件。为了实现这一目标，最近开发了几种方法来促进 PrP (Sc) 的灵敏和精确检测，即蛋白质错误折叠循环扩增、构象依赖性免疫分析、解离增强镧系元素荧光免疫分析、毛细管凝胶电泳、荧光相关光谱、流式细胞术此外，还研究了功能相关的朊病毒敏感细胞培养模型，该模型认识到朊病毒感染机制的复杂性，而不是不仅与诊断有关，而且与朊病毒生物学有关。朊病毒蛋白（PrP）基因缺陷的神经元细胞系能够清楚地阐明PrP（C）功能，将有助于理解朊病毒感染机制。在这篇综述中，我们描述了朊病毒疾病诊断方法和细胞培养模型的最新发展趋势及其在朊病毒生物学中的潜在应用。