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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References
Gao R, Cao B, Hu Y, Feng Z, Wang D, Hu W, Chen J, Jie Z, Qiu H, Xu K, Xu X, Lu H, Zhu W, Gao Z, Xiang N, Shen Y, He Z, Gu Y, Zhang Z, Yang Y, Zhao X, Zhou L, Li X, Zou S, Zhang Y, Li X, Yang L, Guo J, Dong J, Li Q, Dong L, Zhu Y, Bai T, Wang S, Hao P, Yang W, Zhang Y, Han J, Yu H, Li D, Gao GF, Wu G, Wang Y, Yuan Z, Shu Y (2013) Human infection with a novel avian-origin influenza A (H7N9) virus. N Engl J Med 368(20):1888–1897. https://doi.org/10.1056/NEJMoa1304459
Li Q, Zhou L, Zhou M, Chen Z, Li F, Wu H, Xiang N, Chen E, Tang F, Wang D, Meng L, Hong Z, Tu W, Cao Y, Li L, Ding F, Liu B, Wang M, Xie R, Gao R, Li X, Bai T, Zou S, He J, Hu J, Xu Y, Chai C, Wang S, Gao Y, Jin L, Zhang Y, Luo H, Yu H, Gao L, Pang X, Liu G, Shu Y, Yang W, Uyeki TM, Wang Y, Wu F, Feng Z (2013) Preliminary report: epidemiology of the avian influenza A (H7N9) outbreak in China. N Engl J Med 370:520–532. https://doi.org/10.1056/NEJMoa1304617
WHO (2013) Overview of the emergence and characteristics of the avian influenza A(H7N9) virus. http://www.who.int/influenza/human_animal_interface/influenza_h7n9/WHO_H7N9_review_31May13pdf
Barnard DL (2009) Animal models for the study of influenza pathogenesis and therapy. Antivir Res 82(2):A110–A122. https://doi.org/10.1016/j.antiviral.2008.12.014
Belser JA, Katz JM, Tumpey TM (2011) The ferret as a model organism to study influenza a virus infection. Dis Model Mech 4(5):575–579. https://doi.org/10.1242/dmm.007823
O’Donnell CD, Subbarao K (2011) The contribution of animal models to the understanding of the host range and virulence of influenza a viruses. Microbes Infect 13(5):502–515. https://doi.org/10.1016/j.micinf.2011.01.014
Tran V, Moser LA, Poole DS, Mehle A (2013) Highly sensitive real-time in vivo imaging of an influenza reporter virus reveals dynamics of replication and spread. J Virol 87(24):13321–13329. https://doi.org/10.1128/JVI.02381-13
Tran V, Poole DS, Jeffery JJ, Sheahan TP, Creech D, Yevtodiyenko A, Peat AJ, Francis KP, You S, Mehle A (2015) Multi-modal imaging with a toolbox of influenza A Reporter viruses. Viruses 7(10):5319–5327. https://doi.org/10.3390/v7102873
Karlsson EA, Hertz T, Johnson C, Mehle A, Krammer F, Schultz-Cherry S (2016) Obesity outweighs protection conferred by Adjuvanted influenza vaccination. MBio 7(4):e01144. https://doi.org/10.1128/mBio.01144-16
Karlsson EA, Meliopoulos VA, Savage C, Livingston B, Mehle A, Schultz-Cherry S (2015) Visualizing real-time influenza virus infection, transmission and protection in ferrets. Nat Commun 6:6378. https://doi.org/10.1038/ncomms7378
Fulton BO, Palese P, Heaton NS (2015) Replication-competent influenza B reporter viruses as tools for screening antivirals and antibodies. J Virol 89(23):12226–12231. https://doi.org/10.1128/jvi.02164-15
Weisshaar M, Cox R, Morehouse Z, Kumar Kyasa S, Yan D, Oberacker P, Mao S, Golden JE, Lowen AC, Natchus MG, Plemper RK (2016) Identification and characterization of influenza virus entry inhibitors through dual Myxovirus high-throughput screening. J Virol 90(16):7368–7387. https://doi.org/10.1128/jvi.00898-16
Yan D, Weisshaar M, Lamb K, Chung HK, Lin MZ, Plemper RK (2015) Replication-competent influenza virus and respiratory syncytial virus luciferase reporter strains engineered for co-infections identify antiviral compounds in combination screens. Biochemistry 54(36):5589–5604. https://doi.org/10.1021/acs.biochem.5b00623
Hall MP, Unch J, Binkowski BF, Valley MP, Butler BL, Wood MG, Otto P, Zimmerman K, Vidugiris G, Machleidt T, Robers MB, Benink HA, Eggers CT, Slater MR, Meisenheimer PL, Klaubert DH, Fan F, Encell LP, Wood KV (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol 7(11):1848–1857. https://doi.org/10.1021/cb3002478
Matsuoka Y, Lamirande EW, Subbarao K (2009) The mouse model for influenza. Current protocols in microbiology Chapter 15:Unit 15G.13. https://doi.org/10.1002/9780471729259.mc15g03s13
Matsuoka Y, Lamirande EW, Subbarao K (2009) The ferret model for influenza. Current protocols in microbiology Chapter 15:Unit 15G.12. https://doi.org/10.1002/9780471729259.mc15g02s13
Balish AL, Katz JM, Klimov AI (2013) Influenza: propagation, quantification, and storage. Current protocols in microbiology Chapter 15:Unit 15G.11. https://doi.org/10.1002/9780471729259.mc15g01s29
Reed L, Muench H (1938) A simple method of estimating fifty percent endpoints. Am J Epidemiol 27:493–497
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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