Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
J Virol. 2013 Dec;87(24):13321-9. doi: 10.1128/JVI.02381-13. Epub 2013 Oct 2.
The continual public health threat posed by the emergence of novel influenza viruses necessitates the ability to rapidly monitor infection and spread in experimental systems. To analyze real-time infection dynamics, we have created a replication-competent influenza reporter virus suitable for in vivo imaging. The reporter virus encodes the small and bright NanoLuc luciferase whose activity serves as an extremely sensitive readout of viral infection. This virus stably maintains the reporter construct and replicates in culture and in mice with near-native properties. Bioluminescent imaging of the reporter virus permits serial observations of viral load and dissemination in infected animals, even following clearance of a sublethal challenge. We further show that the reporter virus recapitulates known restrictions due to host range and antiviral treatment, suggesting that this technology can be applied to studying emerging influenza viruses and the impact of antiviral interventions on infections in vivo. These results describe a generalizable method to quickly determine the replication and pathogenicity potential of diverse influenza strains in animals.
新型流感病毒的不断出现对公众健康构成持续威胁，这就需要能够快速监测实验系统中的感染和传播情况。为了分析实时感染动态，我们构建了一种具有复制能力的流感报告病毒，该病毒适用于体内成像。报告病毒编码的小型明亮的 NanoLuc 荧光素酶的活性可作为病毒感染的极其敏感的读数。该病毒稳定地维持报告基因构建体，并在培养物和具有近乎天然特性的小鼠中复制。报告病毒的生物发光成像可允许对受感染动物中的病毒载量和传播进行连续观察，甚至在亚致死性挑战后清除后也能进行观察。我们进一步表明，报告病毒再现了由于宿主范围和抗病毒治疗而导致的已知限制，这表明该技术可用于研究新兴流感病毒以及抗病毒干预措施对体内感染的影响。这些结果描述了一种可快速确定不同流感株在动物中的复制和致病性潜力的通用方法。
