Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
Sci Rep. 2018 Jul 12;8(1):10491. doi: 10.1038/s41598-018-28890-0.
ZIKV has emerged as a significant human pathogene for the severe neurological complications, including Guillain-Barré(GBS) syndrome in adults and a variety of fetal abnormalities such as microcephaly. A stable and efficient infectious clone of Brazilian ZIKV isolate is required to study pathogenesis of epidemic ZIKV and virus evolution impact on it. Here we successfully constructed infectious cDNA clone on an early Brazilian isolate by eliminating the activity of predicted bacterial promoter in 1-3000 nt of ZIKV genome, leading to a stable infectious cDNA clone (pZL1). pZL1 derived virus could infect different cell lines and cause lethal effect to AG6 mice. We further investigated the role of a recent emerged substitution in NS5 (M2634V). We found that a reverse mutation (V2634M) caused negligible effect on the ZIKV viral genome replication and infectious progeny production in multiple cell culture systems. Additionally, this mutation did not alter the pathogenesis feature and virulence of ZIKV in AG6 mice. In summary, our results present another robust infectious ZIKV clone from Brazilian isolate and provide evidences to support that M2634V single mutation did not alter virus life cycle in cell culture and pathogenesis in AG6 mouse model.
寨卡病毒已成为导致严重神经并发症的重要人类病原体，包括成人吉兰-巴雷（GBS）综合征和各种胎儿异常，如小头畸形。需要稳定且高效的巴西寨卡病毒分离株感染性克隆来研究流行寨卡病毒的发病机制以及病毒对其进化的影响。在这里，我们通过消除寨卡病毒基因组 1-3000 位核苷酸中预测的细菌启动子的活性，成功构建了早期巴西分离株的感染性 cDNA 克隆，导致稳定的感染性 cDNA 克隆（pZL1）。pZL1 衍生的病毒可以感染不同的细胞系，并导致 AG6 小鼠致死。我们进一步研究了最近出现的 NS5（M2634V）取代的作用。我们发现，在多种细胞培养系统中，反向突变（V2634M）对寨卡病毒基因组复制和感染性后代产生几乎没有影响。此外，该突变并未改变寨卡病毒在 AG6 小鼠中的发病机制特征和毒力。总之，我们的结果提供了另一个来自巴西分离株的强大的感染性寨卡病毒克隆，并提供了证据支持 M2634V 单点突变并未改变病毒在细胞培养中的生命周期和 AG6 小鼠模型中的发病机制特征。
