Pagano Nathan, Aguilar Perez Gerard, Garcia-Milian Rolando, Manuelidis Laura
Section of Neuropathology Surgery, Yale University Medical School, New Haven, Connecticut, United States of America.
Bioinformatics Support Hub, Yale Medical Library, Yale School of Medicine, New Haven, Connecticut, United States of America.
PLoS One. 2025 May 28;20(5):e0323825. doi: 10.1371/journal.pone.0323825. eCollection 2025.
Rat post-mitotic septal neurons, engineered to reversibly proliferate and arrest under physiological conditions, can be maintained for weeks without cytotoxic effects. Nine representative independent cDNA libraries were made to evaluate global arrest-induced neural differentiation and innate immune responses, e.g., upregulated interferon (β-IFN) RNA, that were previously identified in normal uninfected and Creutzfeldt-Jakob Disease agent (CJ) infected septal neurons. This reversible cell model encompassed a non-productive latent (CJ-) and a highly infectious (CJ + , 10 logs/gm) state. Arrest of normal uninfected neurons upregulated a plethora of anti-proliferative transcripts and known neuronal differentiation transcripts (e.g., Neuregulin-1, GDF6 and Prnp). As expected, many activated IFN innate immune genes were simultaneously upregulated (e.g., OAS1, ISG20, CD80, cytokines, chemokines and complement) along with clusterin (CLU) that binds misfolded proteins. Arrest of latently infected CJ- cells induced even more profound global transcript differences. CJ+ cells markedly downregulated the anti-proliferative controls seen in arrested normal cells. CJ+ infection also suppressed neuronal differentiation transcripts, including Prnp which is essential for CJ infection. In contrast, IFN and cytokine/chemokine pathways were strongly upregulated. 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 enhances and expands anti-viral responses; 3) non-productive latent infection can epigenetically imprint many proliferative pathways to thwart complete arrest. This rare cell model of latent infection is fundamental for interrogating triggers of late onset disease that are also relevant for Alzheimer's Disease. Peripheral human blood and intestinal myeloid cells that are latently infected may also be conditionally stimulated in vitro to produce CJ+ linked diagnostic transcripts.
经过基因改造的大鼠有丝分裂后隔区神经元，能够在生理条件下可逆地增殖和停滞，可维持数周而无细胞毒性作用。构建了9个具有代表性的独立cDNA文库，以评估整体停滞诱导的神经分化和先天免疫反应，例如先前在未感染的正常和感染克雅氏病病原体（CJ）的隔区神经元中鉴定出的上调干扰素（β-干扰素）RNA。这种可逆细胞模型包括非生产性潜伏（CJ-）和高传染性（CJ+，10对数/克）状态。正常未感染神经元的停滞上调了大量抗增殖转录本和已知的神经分化转录本（例如神经调节蛋白-1、生长分化因子6和朊蛋白）。正如预期的那样，许多激活的干扰素先天免疫基因以及与错误折叠蛋白结合的簇集蛋白（CLU）同时上调（例如2'-5'-寡腺苷酸合成酶1、干扰素刺激基因20、CD80、细胞因子、趋化因子和补体）。潜伏感染的CJ-细胞的停滞诱导了更深刻的整体转录差异。CJ+细胞显著下调了停滞正常细胞中可见的抗增殖控制。CJ+感染还抑制了神经分化转录本，包括对CJ感染至关重要的朊蛋白。相比之下，干扰素和细胞因子/趋化因子途径强烈上调。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，例如白细胞介素17、干扰素刺激基因15和维甲酸诱导基因I（蝰蛇毒素）。这些数据表明：1）正常神经元在分化过程中产生先天免疫转录本；2）CJ感染增强并扩大抗病毒反应；3）非生产性潜伏感染可在表观遗传上印记许多增殖途径以阻碍完全停滞。这种罕见的潜伏感染细胞模型对于探究与阿尔茨海默病相关的迟发性疾病触发因素至关重要。潜伏感染的外周人血和肠道髓样细胞也可能在体外受到条件刺激，以产生与CJ+相关的诊断转录本。
