Butler D A, Scott M R, Bockman J M, Borchelt D R, Taraboulos A, Hsiao K K, Kingsbury D T, Prusiner S B
Department of Neurology, University of California, San Francisco 94143-0518.
J Virol. 1988 May;62(5):1558-64. doi: 10.1128/JVI.62.5.1558-1564.1988.
Scrapie and Creutzfeldt-Jakob disease are transmissible, degenerative neurological diseases caused by prions. Considerable evidence argues that prions contain protease-resistant sialoglycoproteins, designated PrPSc, encoded by a cellular gene. The prion protein (PrP) gene also encodes a normal cellular protein designated PrPC. We established clonal cell lines which support the replication of mouse scrapie or Creutzfeldt-Jakob disease prions. Mouse neuroblastoma N2a cells were exposed to mouse scrapie prions and subsequently cloned. After limited proteinase K digestion, three PrP-immunoreactive proteins with apparent molecular masses ranging between 20 and 30 kilodaltons were detected in extracts of scrapie-infected N2a cells by Western (immuno-) blotting. The authenticity of these PrPSc molecules was established by using monospecific antiserum raised against a synthetic peptide corresponding to a portion of the prion protein. Those clones synthesizing PrPSc molecules possessed scrapie prion infectivity as measured by bioassay; clones without PrPSc failed to demonstrate infectivity. Detection of PrPSc molecules in scrapie-infected N2a cells supports the contention that PrPSc is a component of the infectious scrapie particle and opens new approaches to the study of prion diseases.
羊瘙痒症和克雅氏病是由朊病毒引起的可传播的退行性神经疾病。大量证据表明，朊病毒含有由细胞基因编码的抗蛋白酶唾液糖蛋白，称为PrPSc。朊病毒蛋白（PrP）基因还编码一种正常的细胞蛋白，称为PrPC。我们建立了支持小鼠羊瘙痒症或克雅氏病朊病毒复制的克隆细胞系。将小鼠神经母细胞瘤N2a细胞暴露于小鼠羊瘙痒症朊病毒，随后进行克隆。经过有限的蛋白酶K消化后，通过蛋白质免疫印迹法在羊瘙痒症感染的N2a细胞提取物中检测到三种表观分子量在20至30千道尔顿之间的PrP免疫反应性蛋白。通过使用针对与朊病毒蛋白一部分相对应的合成肽产生的单特异性抗血清，确定了这些PrPSc分子的真实性。通过生物测定法测量，那些合成PrPSc分子的克隆具有羊瘙痒症朊病毒感染性；没有PrPSc的克隆没有表现出感染性。在羊瘙痒症感染的N2a细胞中检测到PrPSc分子支持了PrPSc是传染性羊瘙痒症颗粒的一个组成部分的观点，并为朊病毒疾病的研究开辟了新途径。
