Thunuguntla Prasanth, El-Mayet Fouad S, Jones Clinton
Oklahoma State University, Center for Veterinary Health Sciences, Department of Veterinary Pathobiology, Stillwater, OK 74078, United States.
Oklahoma State University, Center for Veterinary Health Sciences, Department of Veterinary Pathobiology, Stillwater, OK 74078, United States; Benha University, Faculty of Veterinary Medicine, Department of Virology, Moshtohor 13736, Kaliobyia, Egypt.
Virus Res. 2017 Mar 15;232:1-5. doi: 10.1016/j.virusres.2017.01.011. Epub 2017 Jan 16.
Bovine herpesvirus 1 (BoHV-1) is a significant bovine pathogen that establishes a life-long latent infection in sensory neurons. Previous attempts to develop immortalized bovine neuronal cells were unsuccessful. Consequently, our understanding of the BoHV-1 latency-reactivation cycle has relied on studying complex virus-host interactions in calves. In this study, we tested whether BoHV-1 can infect human (SH-SY5Y) or mouse (Neuro-2A) neuroblastoma cells. We provide new evidence that BoHV-1 efficiently infects SH-SY5Y cells and yields virus titers approximately 100 fold less than bovine kidney cells. Conversely, virus titers from productively infected Neuro-2A cells were approximately 10,000 fold less than bovine kidney cells. Using a β-Gal expressing virus (gC-Blue), we demonstrate that infection of Neuro-2A cells (actively dividing or differentiated) does not result in efficient virus spread, unlike bovine kidney or SH-SY5Y cells. Additional studies demonstrated that lytic cycle viral gene expression (bICP4 and gE) was readily detected in SH-SY5Y cells: conversely bICP4 was not readily detected in productively infected Neuro-2A cells. Finally, infection of SH-SY5Y and bovine kidney cells, but not Neuro-2A cells, led to rapid activation of the Akt protein kinase. These studies suggest that the Neuro-2A cell line may be a novel cell culture model to identify factors that regulate BoHV-1 productive infection in neuronal cells.
牛疱疹病毒1型（BoHV-1）是一种重要的牛病原体，可在感觉神经元中建立终身潜伏感染。此前尝试建立永生化牛神经元细胞的努力均未成功。因此，我们对BoHV-1潜伏-激活周期的理解一直依赖于研究犊牛体内复杂的病毒-宿主相互作用。在本研究中，我们测试了BoHV-1是否能感染人神经母细胞瘤细胞（SH-SY5Y）或小鼠神经母细胞瘤细胞（Neuro-2A）。我们提供了新的证据，表明BoHV-1能有效感染SH-SY5Y细胞，产生的病毒滴度比牛肾细胞低约100倍。相反，经有效感染的Neuro-2A细胞产生的病毒滴度比牛肾细胞低约10000倍。使用表达β-半乳糖苷酶的病毒（gC-Blue），我们证明，与牛肾细胞或SH-SY5Y细胞不同，感染Neuro-2A细胞（活跃分裂或分化状态）不会导致病毒有效传播。进一步的研究表明，在SH-SY5Y细胞中可轻易检测到裂解周期病毒基因表达（bICP4和gE）；相反，在经有效感染的Neuro-2A细胞中则不易检测到bICP4。最后，感染SH-SY5Y细胞和牛肾细胞会导致Akt蛋白激酶快速激活，但感染Neuro-2A细胞则不会。这些研究表明，Neuro-2A细胞系可能是一种新型细胞培养模型，可用于鉴定调节BoHV-1在神经元细胞中进行有效感染的因子。
