Abstract
Reporter viruses are useful probes for studying multiple stages of the viral life cycle. Here we describe an expanded toolbox of fluorescent and bioluminescent influenza A reporter viruses. The enhanced utility of these tools enabled kinetic studies of viral attachment, infection, and co-infection. Multi-modal bioluminescence and positron emission tomography–computed tomography (PET/CT) imaging of infected animals revealed that antiviral treatment reduced viral load, dissemination, and inflammation. These new technologies and applications will dramatically accelerate in vitro and in vivo influenza virus studies.
Highlights
Influenza replication is a highly dynamic process
Existing influenza reporter viruses encoding luciferases or fluorescent proteins have proven useful in rapidly measuring viral infection in vitro and most recently in animals [1,2,3,4,5,6,7,8,9,10,11,12,13,14]
PA-based reporter viruses were rescued as described and incorporation of full-length reporter genes was verified by reverse transcription polymerase chain reaction (RT-PCR) of viral stocks [1,17,20]
Summary
Influenza replication is a highly dynamic process. Yet, the majority of our knowledge about the replication cycle is extrapolated from static snapshots during an infection. This limitation can be overcome by using reporter viruses that facilitate real-time longitudinal measures of virus replication. Existing influenza reporter viruses encoding luciferases or fluorescent proteins have proven useful in rapidly measuring viral infection in vitro and most recently in animals [1,2,3,4,5,6,7,8,9,10,11,12,13,14].
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