Children's Hospital, University of Iowa, Iowa City, Iowa, USA.
J Virol. 2013 Sep;87(17):9643-8. doi: 10.1128/JVI.01506-13. Epub 2013 Jun 26.
Highly pure (>95%) terminally differentiated neurons derived from pluripotent stem cells appear healthy at 2 weeks after infection with varicella-zoster virus (VZV), and the cell culture medium contains no infectious virus. Analysis of the healthy-appearing neurons revealed VZV DNA, transcripts, and proteins corresponding to the VZV immediate early, early, and late kinetic phases of replication. Herein, we further characterized virus in these neuronal cells, focusing on (i) transcription and expression of late VZV glycoprotein C (gC) open reading frame 14 (ORF14) and (ii) ultrastructural features of virus particles in neurons. The analysis showed that gC was not expressed in most infected neurons and gC expression was markedly reduced in a minority of VZV-infected neurons. In contrast, expression of the early-late VZV gE glycoprotein (ORF68) was abundant. Transcript analysis also showed decreased gC transcription compared with gE. Examination of viral structure by high-resolution transmission electron microscopy revealed fewer viral particles than typically observed in cells productively infected with VZV. Furthermore, viral particles were more aberrant, in that most capsids in the nuclei lacked a dense core and most enveloped particles in the cytoplasm were light particles (envelopes without capsids). Together, these results suggest a considerable deficiency in late-phase replication and viral assembly during VZV infection of neurons in culture.
从多能干细胞分化而来的高纯度 (>95%) 终末分化神经元在感染水痘带状疱疹病毒 (VZV) 2 周后看起来健康,且细胞培养液中不含感染性病毒。对外观健康的神经元进行分析发现,VZV DNA、转录本和蛋白质与 VZV 立即早期、早期和晚期复制动力学阶段相对应。在此,我们进一步对这些神经元细胞中的病毒进行了特征分析,重点关注 (i) 晚期 VZV 糖蛋白 C (gC) 开放阅读框 14 (ORF14) 的转录和表达,以及 (ii) 神经元中病毒粒子的超微结构特征。分析表明,大多数感染神经元中未表达 gC,少数 VZV 感染神经元中 gC 表达明显减少。相比之下,早期-晚期 VZV gE 糖蛋白 (ORF68) 的表达丰富。转录分析还显示 gC 转录较 gE 减少。通过高分辨率透射电子显微镜检查病毒结构发现,与 VZV 有效感染细胞中通常观察到的相比,病毒颗粒较少。此外,病毒颗粒更异常,即大多数核内衣壳缺乏致密核心,大多数细胞质中包膜颗粒为轻颗粒(无衣壳的包膜)。这些结果共同表明,在 VZV 感染培养神经元时,晚期复制和病毒装配存在相当大的缺陷。