Lynch W P, Snyder E Y, Qualtiere L, Portis J L, Sharpe A H
Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.
J Virol. 1996 Dec;70(12):8896-907. doi: 10.1128/JVI.70.12.8896-8907.1996.
CasBrE is a neurovirulent murine retrovirus which induces a spongiform myeloencephalopathy in susceptible mice. Genetic mapping studies have indicated that sequences responsible for neurovirulence reside within the env gene. To address the question of direct envelope protein neuroxicity in the central nervous system (CNS), we have generated chimeric mice expressing the CasBrE envelope protein in cells of neuroectodermal origin. Specifically, the multipotent neural progenitor cell line C17.2 was engineered to express the CasBrE env gene as either gp70/p15E (CasE) or gp70 alone (CasES). CasE expression in these cells resulted in complete (>10(5)) interference of superinfection with Friend murine leukemia virus clone FB29, whereas CasES expression resulted in a 1.8-log-unit decrease in FB29 titer. Introduction of these envelope-expressing C17.2 cells into the brains of highly susceptible IRW mice resulted in significant engraftment as integral cytoarchitecturally correct components of the CNS. Despite high-level envelope protein expression from the engrafted cells, no evidence of spongiform neurodegeneration was observed. To examine whether early virus replication events were necessary for pathogenesis, C17.2 cells expressing whole virus were transplanted into mice in which virus replication in the host was specifically restricted by Fv-1 to preintegration events. Again, significant C17.2 cell engraftment and infectious virus expression failed to precipitate spongiform lesions. In contrast, transplantation of virus-expressing C17.2 progenitor cells in the absence of the Fv-1 restriction resulted in extensive spongiform neurodegeneration by 2 weeks postengraftment. Cytological examination indicated that infection had spread beyond the engrafted cells, and in particular to host microglia. Spongiform neuropathology in these animals was directly correlated with CasBrE env expression in microglia rather than expression from neural progenitor cells. These results suggest that the envelope protein of CasBrE is not itself neurotoxic but that virus infectious events beyond binding and fusion in microglia are necessary for the induction of CNS disease.
CasBrE是一种具有神经毒性的鼠逆转录病毒,可在易感小鼠中诱发海绵状脊髓脑病。基因定位研究表明,负责神经毒性的序列位于env基因内。为了解决中枢神经系统(CNS)中包膜蛋白直接神经毒性的问题,我们构建了在神经外胚层来源的细胞中表达CasBrE包膜蛋白的嵌合小鼠。具体而言,多能神经祖细胞系C17.2经过基因工程改造,可表达CasBrE env基因,形式为gp70/p15E(CasE)或仅为gp70(CasES)。这些细胞中CasE的表达导致对Friend鼠白血病病毒克隆FB29的超感染完全(>10⁵)受到干扰,而CasES的表达导致FB29滴度降低1.8个对数单位。将这些表达包膜的C17.2细胞引入高度易感的IRW小鼠大脑中,结果显示它们作为CNS中细胞结构正确的完整组成部分实现了显著的植入。尽管植入细胞高水平表达包膜蛋白,但未观察到海绵状神经变性的迹象。为了研究早期病毒复制事件是否是发病机制所必需的,将表达完整病毒的C17.2细胞移植到宿主中病毒复制因Fv-1而特异性限制在整合前事件的小鼠体内。同样,显著的C17.2细胞植入和感染性病毒表达未能引发海绵状病变。相反,在没有Fv-1限制的情况下移植表达病毒的C17.2祖细胞,在植入后2周导致广泛的海绵状神经变性。细胞学检查表明感染已扩散到植入细胞之外,特别是扩散到宿主小胶质细胞。这些动物中的海绵状神经病理学与小胶质细胞中CasBrE env的表达直接相关,而不是与神经祖细胞的表达相关。这些结果表明,CasBrE的包膜蛋白本身没有神经毒性,但小胶质细胞中除结合和融合之外的病毒感染事件对于诱发CNS疾病是必需的。