Abstract
To streamline standard virological assays, we developed bioluminescent replication-competent influenza reporter viruses that mimic their parental counterparts. These reporter viruses provide a rapid and quantitative readout of viral infection and replication. Moreover, they permit real-time in vivo measures of viral load, tissue distribution, and transmission in the same cohort of animals over the entire course of infection—measurements that were not previously possible. Here we provide detailed protocols using bioluminescent reporter viruses for in vivo imaging in mice and ferrets. We also describe cell culture-based techniques using reporter viruses for quantification of viral titers and performing microneutralization assays. The ease, speed, and adaptability of these approaches have the potential to accelerate multiple areas of influenza virus research.
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Acknowledgments
Funding: This work was supported by the National Institutes of Health-National Institute of Allergies and Infectious Diseases contract numbers HHSN266200700005C and HHSN272201400006C and the American Lebanese Syrian Associated Charities (ALSAC) to S.S.-C. and by the National Institutes of Health-National Institute of General Medical Sciences (R00GM088484), the American Lung Association Basic Research Grant (RG-310016), a Shaw Scientist Award, and a Wisconsin Partnership Education and Research Committee New Investigator Program grant to A.M. V.T. was supported by an NIH National Research Service Award (T32 GM07215). A.M. is a Burroughs Wellcome Fund Investigator in the Pathogenesis of Infectious Disease.
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Karlsson, E.A. et al. (2018). Measuring Influenza Virus Infection Using Bioluminescent Reporter Viruses for In Vivo Imaging and In Vitro Replication Assays. In: Yamauchi, Y. (eds) Influenza Virus. Methods in Molecular Biology, vol 1836. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8678-1_21
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DOI: https://doi.org/10.1007/978-1-4939-8678-1_21
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