Prion strain discrimination in cell culture: The cell panel assay
Prion strain discrimination in cell culture: The cell panel assay
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DOI：
10.1073/pnas.0710054104
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发表时间：
2007-12-26
影响因子：
11.1
通讯作者：
Weissmann, Charles
中科院分区：
文献类型：
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作者：
Mahal, Sukhvir P.;Baker, Christopher A.;Weissmann, Charles
朊病毒被认为主要或完全由错误折叠的 PrP（一种组成型表达的宿主蛋白）组成。与相同 PrP 序列相关的朊病毒可能以不同菌株的形式出现；菌株表型被认为是由 PrP 的构象编码的。一些细胞系可以持续被朊病毒感染，有趣的是，它们对某些菌株表现出偏好。我们报告克隆的小鼠神经母细胞瘤细胞群 N2a-PK1 对 RML 和 22L 朊病毒的敏感性具有高度异质性。值得注意的是，兄弟亚克隆可能对两种菌株表现出非常不同的相对敏感性，表明反应可以独立变化。我们已经将四种细胞系 N2a-PK1、N2a-R33、LD9 和 CAD5 组装成一个组，这些细胞系对朊病毒株 RML、22L、301C 和 Me7 显示出截然不同的反应，可以使用标准瘙痒细胞测定 (SSCA) 在 2 周内进行体外区分。
Prions are thought to consist mainly or entirely of misfolded PrP, a constitutively expressed host protein. Prions associated with the same PrP sequence may occur in the form of different strains; the strain phenotype is believed to be encoded by the conformation of the PrP. Some cell lines can be persistently infected by prions and, interestingly, show preference for certain strains. We report that a cloned murine neuroblastoma cell population, N2a-PK1, is highly heterogeneous in regard to its susceptibility to RML and 22L prions. Remarkably, sibling subclones may show very different relative susceptibilities to the two strains, indicating that the responses can vary independently. We have assembled four cell lines, N2a-PK1, N2a-R33, LD9 and CAD5, which show widely different responses to prion strains RML, 22L, 301C, and Me7, into a panel that allows their discrimination in vitro within 2 weeks, using the standard scrapie cell assay (SSCA).