Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies.
Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies.
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DOI：
10.1016/j.virol.2016.04.020
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发表时间：
2016-07
期刊：
影响因子：
3.7
通讯作者：
Bhatia SN
中科院分区：
文献类型：
--
作者：
Ramanan V;Trehan K;Ong ML;Luna JM;Hoffmann HH;Espiritu C;Sheahan TP;Chandrasekar H;Schwartz RE;Christine KS;Rice CM;van Oudenaarden A;Bhatia SN
丙型肝炎病毒(丙型肝炎病毒)是一种阳性的单链RNA病毒，具有巨大的全球健康重要性，直接作用的抗病毒疗法取代了基于免疫刺激干扰素的疗法。丙型肝炎病毒正链和负链病毒RNA(VRNAs)在抗病毒干扰下的动力学还没有在单细胞水平上进行研究，这使得我们对抗病毒动力学和宿主-病毒相互作用的理解存在差距。在这里，我们展示了多种感染模型中丙型肝炎病毒基因组的定量成像，以及正链和负链vRNAs和宿主抗病毒RNAs的多路复用。我们捕捉了不同作用机制的抗病毒药物清除丙型肝炎病毒感染的不同动力学，发现NS5A抑制剂达拉塔韦可以诱导负链病毒RNA迅速下降。我们还发现，干扰素治疗后宿主抗病毒基因的诱导与单细胞中的病毒载量呈正相关。这项研究将smFISH添加到可用于分析RNA病毒感染治疗的工具箱中。
Hepatitis C virus (HCV) is a positive single-stranded RNA virus of enormous global health importance, with direct-acting antiviral therapies replacing an immunostimulatory interferon-based regimen. The dynamics of HCV positive and negative-strand viral RNAs (vRNAs) under antiviral perturbations have not been studied at the single-cell level, leaving a gap in our understanding of antiviral kinetics and host-virus interactions. Here, we demonstrate quantitative imaging of HCV genomes in multiple infection models, and multiplexing of positive and negative strand vRNAs and host antiviral RNAs. We capture the varying kinetics with which antiviral drugs with different mechanisms of action clear HCV infection, finding the NS5A inhibitor daclatasvir to induce a rapid decline in negative-strand viral RNAs. We also find that the induction of host antiviral genes upon interferon treatment is positively correlated with viral load in single cells. This study adds smFISH to the toolbox available for analyzing the treatment of RNA virus infections.