Accelerated high fidelity prion amplification within and across prion species barriers
Accelerated high fidelity prion amplification within and across prion species barriers
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DOI：
10.1371/journal.ppat.1000139
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发表时间：
2008-08-01
期刊：
影响因子：
6.7
通讯作者：
Telling, Glenn C.
中科院分区：
文献类型：
--
作者：
Green, Kristi M.;Castilla, Joaquin;Telling, Glenn C.
实验上的障碍阻碍了我们研究朊病毒在物种内部，尤其是物种之间传播的能力。在这里，我们使用转基因小鼠脑提取物中表达的宫颈朊病毒蛋白Tg（CerPrP）作为底物，通过蛋白质错误折叠循环扩增（PMCA）在体外产生慢性消耗性疾病（CWD）朊病毒。在Tg（CerPrP）小鼠中对这种传染性的表征表明，连续PMCA导致CWD朊病毒的高保真扩增，其性质明显不变。使用类似的方法扩增小鼠RML朊病毒并表征由此产生的新型宫颈朊病毒，研究人员发现，在Tg（CerPrP）小鼠中，连续PMCA消除了需要数百天适应和随后稳定的传播屏障。虽然这两种方法产生的宫颈朊病毒具有与CWD不同的特征，但所得到的单个朊病毒分离物的细微差异表明，小鼠RML朊病毒的适应性在种间传播后产生了多个毒株。我们的研究表明，转基因小鼠和PMCA结合的方法不仅加快了朊病毒在种内和种间的传播，而且提供了一种简便的方法来产生和表征新的朊病毒菌株。
Experimental obstacles have impeded our ability to study prion transmission within and, more particularly, between species. Here, we used cervid prion protein expressed in brain extracts of transgenic mice, referred to as Tg(CerPrP), as a substrate for in vitro generation of chronic wasting disease (CWD) prions by protein misfolding cyclic amplification (PMCA). Characterization of this infectivity in Tg(CerPrP) mice demonstrated that serial PMCA resulted in the high fidelity amplification of CWD prions with apparently unaltered properties. Using similar methods to amplify mouse RML prions and characterize the resulting novel cervid prions, we show that serial PMCA abrogated a transmission barrier that required several hundred days of adaptation and subsequent stabilization in Tg(CerPrP) mice. While both approaches produced cervid prions with characteristics distinct from CWD, the subtly different properties of the resulting individual prion isolates indicated that adaptation of mouse RML prions generated multiple strains following inter-species transmission. Our studies demonstrate that combined transgenic mouse and PMCA approaches not only expedite intra-and inter-species prion transmission, but also provide a facile means of generating and characterizing novel prion strains.