Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Viruses. 2020 Jan 9;12(1):80. doi: 10.3390/v12010080.
Among the causative agents of duck viral hepatitis, duck hepatitis A virus genotype 1 (DHAV-1) is the most common virus reported in most outbreaks worldwide. How to propagate DHAV-1 in cell cultures efficiently remains a problem to be explored. Here, we aimed to test the effect of serum type on DHAV-1 replication in duck embryo fibroblast (DEF) cells. Comparative studies involved virus culture and passage, observation of cytopathic effect (CPE), virus quantification, and plaque formation assay. From the results of these investigations, we conclude that use of chicken serum (CS) in maintenance medium allows DHAV-1 to establish productive, cytocidal infection in DEF cells, whereas FCS exerts inhibitory effects on DHAV-1 replication, CPE development, and plaque formation. By using a neutralization test, we found that the direct action of FCS on virions is likely to play a key role in inhibiting DHAV-1 replication in DEF cells. Mechanism analyses revealed that FCS inhibits DHAV-1 replication at virus adsorption and reduces extracellular virus yields. The present work may shed light on a new perspective for antiviral agent development, and have provided a virus-host cell system for further studies on molecular mechanism involved DHAV-1 replication and pathogenesis.
在鸭病毒性肝炎的病原体中,鸭甲型肝炎病毒 1 型(DHAV-1)是世界范围内大多数暴发中最常见的病毒。如何在细胞培养中有效地繁殖 DHAV-1 仍然是一个有待探索的问题。在这里,我们旨在测试血清型对鸭胚成纤维细胞(DEF)中 DHAV-1 复制的影响。比较研究涉及病毒培养和传代、观察细胞病变效应(CPE)、病毒定量和噬斑形成测定。从这些研究的结果中,我们得出结论,在维持培养基中使用鸡血清(CS)可使 DHAV-1 在 DEF 细胞中建立有生产力、细胞杀伤性感染,而 FCS 对 DHAV-1 复制、CPE 发展和噬斑形成具有抑制作用。通过中和试验,我们发现 FCS 对病毒粒子的直接作用可能在抑制 DEF 细胞中 DHAV-1 的复制中起关键作用。机制分析表明,FCS 在病毒吸附时抑制 DHAV-1 复制并减少细胞外病毒产量。本工作可能为抗病毒药物的开发提供新的视角,并为进一步研究 DHAV-1 复制和发病机制提供了病毒-宿主细胞系统。