Zhang Qiu-Yan, Li Xiao-Dan, Liu Si-Qing, Deng Cheng-Lin, Zhang Bo, Ye Han-Qing
Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Arch Virol. 2017 Nov;162(11):3417-3423. doi: 10.1007/s00705-017-3508-9. Epub 2017 Aug 4.
Japanese encephalitis virus (JEV), an important pathogen in Eastern and Southern Asia and the Pacific, has spread to Australia and other territories in recent years. Although the vaccine for JEV has been used in some countries, development of efficient antiviral drugs is still an urgent requirement. Replicon systems have been widely used in the research of viral replication and antiviral screening for West Nile virus (WNV), yellow fever virus (YFV) and dengue virus (DENV). Here, a novel JEV replicon harboring the Rluc and Pac gene (JEV-Pac-Rluc-Rep) was constructed. Furthermore, we established a BHK-21 cell line harboring JEV-Pac-Rluc-Rep (BHK-21 cell line) through continuous puromycin selection. Characterization of cell line stability showed that the replicon RNA could persistently replicate in this cell line for at least up to 10 rounds of passage. Using a known flavivirus inhibitor, the JEV replicon cell line was validated for antiviral screening. The JEV replicon cell line will be a valuable tool for both compound screening and viral replication studies.
日本脑炎病毒（JEV）是东亚、南亚及太平洋地区的一种重要病原体，近年来已传播至澳大利亚及其他地区。尽管JEV疫苗已在一些国家使用，但开发高效抗病毒药物仍是一项迫切需求。复制子系统已广泛应用于西尼罗河病毒（WNV）、黄热病病毒（YFV）和登革病毒（DENV）的病毒复制研究及抗病毒筛选。在此，构建了一种携带Rluc和Pac基因的新型JEV复制子（JEV-Pac-Rluc-Rep）。此外，通过持续的嘌呤霉素筛选，我们建立了一种携带JEV-Pac-Rluc-Rep的BHK-21细胞系（BHK-21细胞系）。细胞系稳定性表征显示，复制子RNA可在该细胞系中持续复制至少10代。使用一种已知的黄病毒抑制剂，对JEV复制子细胞系进行了抗病毒筛选验证。JEV复制子细胞系将成为化合物筛选和病毒复制研究的宝贵工具。
