Tsetsarkin Konstantin A, Kenney Heather, Chen Rubing, Liu Guangping, Manukyan Hasmik, Whitehead Stephen S, Laassri Majid, Chumakov Konstantin, Pletnev Alexander G
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Department of Pathology, Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, Texas, USA.
mBio. 2016 Aug 23;7(4):e01114-16. doi: 10.1128/mBio.01114-16.
An arthropod-borne virus, Zika virus (ZIKV), has recently emerged as a major human pathogen. Associated with complications during perinatal development and Guillain-Barré syndrome in adults, ZIKV raises new challenges for understanding the molecular determinants of flavivirus pathogenesis. This underscores the necessity for the development of a reverse genetic system based on an epidemic ZIKV strain. Here, we describe the generation and characterization in cell cultures of an infectious cDNA clone of ZIKV isolated from the 2015 epidemic in Brazil. The cDNA-derived ZIKV replicated efficiently in a variety of cell lines, including those of both neuronal and placental origin. We observed that the growth of cDNA-derived virus was attenuated compared to the growth of the parental isolate in most cell lines, which correlates with substantial differences in sequence heterogeneity between these viruses that were determined by deep-sequencing analysis. Our findings support the role of genetic diversity in maintaining the replicative fitness of viral populations under changing conditions. Moreover, these results indicate that caution should be exercised when interpreting the results of reverse-genetics experiments in attempts to accurately predict the biology of natural viruses. Finally, a Vero cell-adapted cDNA clone of ZIKV was generated that can be used as a convenient platform for studies aimed at the development of ZIKV vaccines and therapeutics.
The availability of genetic tools and laboratory models determines the progress in understanding mechanisms of virus emergence and pathogenesis. Recent large-scale outbreaks of Zika virus (ZIKV) that were linked to complications during perinatal development and Guillain-Barré syndrome in adults emphasize the urgency for the development of a reverse-genetics system based on an epidemic ZIKV strain. Here, we report a stable infectious cDNA clone for ZIKV isolated during the 2015 epidemic in Brazil, as well as a Vero cell-adapted version of it, which will be used for virus-host interaction studies and vaccine development.
一种节肢动物传播的病毒——寨卡病毒（ZIKV），最近已成为一种主要的人类病原体。寨卡病毒与围产期发育并发症及成人吉兰 - 巴雷综合征相关，这为理解黄病毒发病机制的分子决定因素带来了新挑战。这凸显了基于流行的寨卡病毒株开发反向遗传系统的必要性。在此，我们描述了从2015年巴西疫情中分离出的寨卡病毒感染性cDNA克隆在细胞培养中的构建及特性。该cDNA衍生的寨卡病毒能在多种细胞系中高效复制，包括神经源性和胎盘源性细胞系。我们观察到，在大多数细胞系中，与亲本病毒分离株相比，cDNA衍生病毒的生长有所减弱，这与通过深度测序分析确定的这些病毒在序列异质性上的显著差异相关。我们的研究结果支持了遗传多样性在不断变化的条件下维持病毒群体复制适应性中的作用。此外，这些结果表明，在解释反向遗传学实验结果以准确预测天然病毒生物学特性时应谨慎。最后，构建了一种适应Vero细胞的寨卡病毒cDNA克隆，可作为开发寨卡病毒疫苗和治疗方法研究的便利平台。
遗传工具和实验室模型的可用性决定了在理解病毒出现和发病机制方面的进展。近期与围产期发育并发症及成人吉兰 - 巴雷综合征相关的寨卡病毒（ZIKV）大规模疫情凸显了基于流行的寨卡病毒株开发反向遗传系统的紧迫性。在此，我们报告了一株2015年巴西疫情期间分离的寨卡病毒稳定感染性cDNA克隆及其适应Vero细胞的版本，这将用于病毒 - 宿主相互作用研究和疫苗开发。
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