Single-Cell Virology: On-Chip Investigation of Viral Infection Dynamics.
Single-Cell Virology: On-Chip Investigation of Viral Infection Dynamics.
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DOI：
10.1016/j.celrep.2017.10.051
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发表时间：
2017-11-07
期刊：
影响因子：
8.8
通讯作者：
Cameron CE
中科院分区：
文献类型：
--
作者：
Guo F;Li S;Caglar MU;Mao Z;Liu W;Woodman A;Arnold JJ;Wilke CO;Huang TJ;Cameron CE
我们已经开发了一个基于微流控的高通量平台，用于对单个细胞中的病毒感染进行动力学分析。我们分析了数以千计的脊髓灰质炎病毒感染个体，同时分析了不同的实验参数，包括感染的多样性、细胞周期、病毒基因型和药物的存在。我们做出了几个被基于群体的实验掩盖的意想不到的观察：(1)病毒和细胞因素对病毒感染动力学做出独特而独立的贡献；(2)细胞因素导致复制开始时间的广泛变化；(3)感染通常开始得更晚，复制发生得比人口测量预测的更快。我们发现，突变负荷损害了病毒种群与宿主的相互作用，延迟了复制开始时间，并解释了突变子病毒的减弱表型。我们证明了一种抗病毒药物可以选择性地消灭最适合病毒群体的成员。单细胞病毒学有助于发现和表征毒力决定因素，并有助于阐明群体方法所不能理解的药物作用机制。郭某等人。使用安装在荧光显微镜上的微流体设备来监测单个细胞中病毒感染的动力学。感染结局的细胞间差异存在于生命周期的所有阶段。细胞基因表达决定了月食阶段；病毒遗传决定了复制速度和产量。
We have developed a high-throughput, microfluidics-based platform to perform kinetic analysis of viral infections in individual cells. We have analyzed thousands of individual poliovirus infections while varying experimental parameters, including multiplicity of infection, cell cycle, viral genotype and presence of a drug. We make several unexpected observations masked by population-based experiments: (1) viral and cellular factors contribute uniquely and independently to viral infection kinetics; (2) cellular factors cause wide variation in replication-start times; (3) infections frequently begin later and replication occurs faster than predicted by population measurements. We show that mutational load impairs interaction of the viral population with the host, delaying replication start times and explaining the attenuated phenotype of a mutator virus. We show that an antiviral drug can selectively extinguish the most-fit members of the viral population. Single-cell virology facilitates discovery and characterization of virulence determinants and elucidation of mechanisms of drug action eluded by population methods. Guo et al. use a microfluidics device installed on a fluorescence microscope to monitor the kinetics of viral infection in single cells. Between-cell variation in outcomes of infection exist at all phases of the lifecycle. Cellular gene expression governs the eclipse phase; viral genetics govern replication rate and yield.
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