Successful transmission of three mouse-adapted scrapie strains to murine neuroblastoma cell lines overexpressing wild-type mouse prion protein
Successful transmission of three mouse-adapted scrapie strains to murine neuroblastoma cell lines overexpressing wild-type mouse prion protein
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DOI：
10.1128/jvi.74.1.320-325.2000
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发表时间：
2000-01-01
影响因子：
5.4
通讯作者：
Lehmann, S
中科院分区：
文献类型：
--
作者：
Nishida, N;Harris, DA;Lehmann, S
传播性海绵状脑病（TSE）的代理人在培养的细胞中的繁殖已经实现了只有少数细胞系。为了建立有效和通用的传播模型，我们开发了过表达A型小鼠朊病毒蛋白MoPrPC-A的神经母细胞瘤细胞系，然后测试了细胞对几种不同的小鼠适应性瘙痒病株的易感性。转染的细胞克隆表达高达6倍的PrPC水平比未转染的细胞。即使在30次传代后，我们也能够在试剂接种的PrP过表达细胞中检测到朊病毒蛋白的异常蛋白酶R抗性形式PrPSc，而在3次传代后未转染的细胞中检测不到PrPSc。这些细胞中PrPSc的产生也比在羊瘙痒病感染的神经母细胞瘤细胞（ScN 2a）中观察到的更高且更稳定。转染细胞仅对PrPSc-A菌株钱德勒、139 A和22 L敏感，而对PrPSc-B菌株87 V和22 A不敏感。我们进一步证明了PrPSc从感染细胞到其他未感染细胞的成功传输。我们的研究结果证实了这一假设，即成功的传播代理离体依赖于宿主PrPC的表达水平和PrPSc的序列。这种新的离体传播模型将促进对宿主与病原体相互作用机制的研究，例如物种屏障和菌株多样性，并为开发可用于诊断和治疗传染性海绵状脑病的高度易感细胞系提供基础。
Propagation of the agents responsible for transmissible spongiform encephalopathies (TSEs) in cultured cells las been achieved for only a few cell lines. To establish efficient and versatile models for transmission, we developed neuroblastoma cell lines overexpressing type A mouse prion protein, MoPrPC-A, and then tested the susceptibility of the cells to several different mouse-adapted scrapie strains. The transfected cell clones expressed up to sixfold-higher levels of PrPC than the untransfected cells. Even after 30 passages, we were able to detect an abnormal proteinase R-resistant form of prion protein, PrPSc, in the agent-inoculated PrP-overexpressing cells, while no PrPSc was detectable in the untransfected cells after 3 passages. Production of PrPSc in these cells was also higher and more stable than that seen in scrapie-infected neuroblastoma cells (ScN2a). The transfected cells mere susceptible to PrPSc-A strains Chandler, 139A, and 22L but not to PrPSc-B strains 87V and 22A. We further demonstrate the successful transmission of PrPSc from infected cells to other uninfected cells. Our results corroborate the hypothesis that the successful transmission of agents ex vivo depends on both expression Levels of host PrPC and the sequence of PrPSc. This new ex vivo transmission model will facilitate research into the mechanism of host-agent interactions, such as the species barrier and strain diversity, and provides a basis for the development of highly susceptible cell lines that could be used in diagnostic and therapeutic approaches to the TSEs.