Two Creutzfeldt-Jakob disease agents reproduce prion protein-independent identities in cell cultures
Two Creutzfeldt-Jakob disease agents reproduce prion protein-independent identities in cell cultures
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DOI：
10.1073/pnas.0400158101
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发表时间：
2004-06-08
影响因子：
11.1
通讯作者：
Manuelidis, L
中科院分区：
文献类型：
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作者：
Arjona, A;Simarro, L;Manuelidis, L
人类克雅氏病（CJD）和类似的神经退行性疾病，如羊瘙痒症是由各种相关的感染因子引起的。它们与异常的宿主朊病毒蛋白（PrP）相关，通过有限的蛋白水解产生抗性PrP条带（PrP-res）进行评估。虽然PrP-res被假定为感染因子，但纯化的PrP-res本身不具有感染性。为了建立独立的CJD剂的特点，从那些PrP-res，两个不同的小鼠传代CJD株繁殖的神经元细胞系，其PrP-res模式显着不同，从那些发现在感染的大脑。在小鼠脑中，快型CJD株FU释放大量PrP-res沉积，而慢型SY株释放很少。这两种菌株诱发PrP-res在培养的小鼠细胞，虽然SY诱导PrP-res只有短暂的。FU和SY感染的GT 1细胞中的PrP-res模式是相同的，并且与脑和N2 a细胞中的PrP-res模式显著不同。尽管如此，所有FU感染的细胞系在小鼠中再现了其原始的快速疾病，即使在广泛的传代培养后，而SY感染的细胞仅产生缓慢的疾病。这些数据表明PrP-res既不编码也不改变试剂特异性特征。PrP-res也是一个很差的感染性预测因子，因为失去PrP-res的SY细胞的感染性比PrP-res阳性培养物高出约10倍。此外，FU滴度增加了650倍，而PrP-res保持不变。经FU感染的传代细胞具有与脑相当的滴度，并且>30%的细胞显示可捕获试剂的丰富的细胞质PrP-res聚集体。CJD在单型细胞中的持续大量复制将进一步区分病原体特异性分子与宿主对感染的病理反应。
Human Creutzfeldt-jakob disease (CJD) and similar neurodegenerative diseases such as sheep scrapie are caused by a variety of related infectious agents. They are associated with abnormal host prion protein (PrP), which is assessed by limited proteolysis to yield resistant PrP bands (PrP-res). Although PrP-res has been posited as the infectious agent, purified PrP-res itself is not infectious. To establish the independence of CJD agent characteristics from those of PrP-res, two different mouse-passaged CJD strains were propagated in neuronal cell lines whose PrP-res patterns differ markedly from each other and from those found in infected brain. In mouse brain, the fast CJD strain, FU, elicits many PrP-res deposits, whereas the slow SY strain elicits few. Both strains evoked PrP-res in cultured murine cells, although SY induced PrP-res only transiently. PrP-res patterns in FU- and SY-infected GT1 cells were identical, and were significantly different from those in brain and in N2a cells. Nevertheless, all FU-infected cell lines reproduced their original fast disease in mice, even after extensive subculture, whereas SY-infected cells produced only slow disease. These data indicate PrP-res neither encodes nor alters agent-specific characteristics. PrP-res was also a poor predictor of infectivity because SY cells that had lost PrP-res were approximate to10-fold more infectious than PrP-res-positive cultures. Furthermore, FU titers increased 650-fold, whereas PrP-res remained constant. Passaged FU-infected cells had titers comparable to brain, and >30% of cells displayed abundant cytoplasmic PrP-res aggregates that may trap agent. The continuous substantial replication of CJD in monotypic cells, will further the discrimination of agent-specific molecules from pathological host responses to infection.