PDF(Pubmed)
Abstract：
Rat post-mitotic septal (SEP) neurons, engineered to conditionally proliferate at 33°C, differentiate when arrested at 37.5°C and can be maintained for weeks without cytotoxic effects. Nine independent cDNA libraries were made to follow arrest-induced neural differentiation and innate immune responses in normal (Nl) uninfected and CJ agent infected SEP cells. Proliferating Nl versus latently infected (CJ-) cells showed few RNA-seq differences. However arrest induced major changes. Normal cells displayed a plethora of anti-proliferative transcripts. Additionally, known neuron differentiation transcripts, e.g., Agtr2, Neuregulin-1, GDF6, SFRP4 and Prnp were upregulated. These Nl neurons also displayed many activated IFN innate immune genes, e.g., OAS1, RTP4, ISG20, GTB4, CD80 and cytokines, complement, and clusterin (CLU) that binds to misfolded proteins. In contrast, arrested highly infectious CJ+ cells (10 logs/gm) downregulated many replication controls. Furthermore, arrested CJ+ cells suppressed neuronal differentiation transcripts, including Prnp which is essential for CJ agent infection. CJ+ cells also enhanced IFN stimulated pathways, and analysis of the 342 CJ+ unique transcripts revealed additional innate immune and anti-viral-linked transcripts, e.g., Il17, ISG15, and RSAD2 (viperin). These data show: 1) innate immune transcripts are produced by normal neurons during differentiation; 2) CJ infection can enhance and expand anti-viral responses; 3) latent CJ infection epigenetically imprints many proliferative pathways to thwart complete arrest. CJ+ brain microglia, white blood cells and intestinal myeloid cells with shared transcripts may be stimulated to educe latent CJD infections that can be clinically silent for >30 years.
摘要：
大鼠有丝分裂后间隔（SEP）神经元，被设计为在33℃有条件地增殖，当在37.5℃下被捕时可以分化，并且可以维持数周而没有细胞毒性作用。制备9个独立的cDNA文库以跟踪正常(N1)未感染和CJ剂感染的SEP细胞中的阻滞诱导的神经分化和先天性免疫应答。增殖的Nl与潜伏感染的(CJ-)细胞显示很少的RNA-seq差异。然而，逮捕引发了重大变化。正常细胞显示过多的抗增殖转录物。此外,已知的神经元分化转录本，例如，Agtr2、神经调节蛋白-1、GDF6、SFRP4和Prnp上调。这些Nl神经元还展示了许多激活的IFN先天免疫基因，例如，OAS1，RTP4，ISG20，GTB4，CD80和细胞因子，补语,和与错误折叠的蛋白质结合的簇蛋白（CLU）。相比之下,被捕的高传染性CJ细胞（10log/gm）下调了许多复制控制。此外，被捕的CJ+细胞抑制了神经元分化转录本，包括对CJ病原体感染至关重要的Prnp。CJ+细胞也增强了IFN刺激途径，对342个CJ+独特转录本的分析揭示了额外的先天免疫和抗病毒相关转录本，例如，Il17、ISG15和RSAD2(viperin)。这些数据显示:1)在分化期间由正常神经元产生先天免疫转录物;2)CJ感染可以增强和扩大抗病毒应答;3)潜伏CJ感染表观遗传印记许多增殖途径以阻止完全停滞。CJ+脑小胶质细胞,具有共享转录本的白细胞和肠道骨髓细胞可能被刺激以产生潜伏的CJD感染，这种感染在临床上可以沉默超过30年。
