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Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins
Eckert, Nadine Wrensch, Florian Gärtner, Sabine Palanisamy, Navaneethan Goedecke, Ulrike Jäger, Nils Pöhlmann, Stefan Winkler, Michael
Eckert, Nadine
Wrensch, Florian
Gärtner, Sabine
Palanisamy, Navaneethan
Goedecke, Ulrike
Jäger, Nils
Pöhlmann, Stefan
Winkler, Michael
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2014-05-19
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Abstract
Reporter genes inserted into viral genomes enable the easy and rapid quantification of virus replication, which is instrumental to efficient in vitro screening of antiviral compounds or in vivo analysis of viral spread and pathogenesis. Based on a published design, we have generated several replication competent influenza A viruses carrying either fluorescent proteins or Gaussia luciferase. Reporter activity could be readily quantified in infected cultures, but the virus encoding Gaussia luciferase was more stable than viruses bearing fluorescent proteins and was therefore analyzed in detail. Quantification of Gaussia luciferase activity in the supernatants of infected culture allowed the convenient and highly sensitive detection of viral spread, and enzymatic activity correlated with the number of infectious particles released from infected cells. Furthermore, the Gaussia luciferase encoding virus allowed the sensitive quantification of the antiviral activity of the neuraminidase inhibitor (NAI) zanamivir and the host cell interferon-inducible transmembrane (IFITM) proteins 1-3, which are known to inhibit influenza virus entry. Finally, the virus was used to demonstrate that influenza A virus infection is sensitive to a modulator of endosomal cholesterol, in keeping with the concept that IFITMs inhibit viral entry by altering cholesterol levels in the endosomal membrane. In sum, we report the characterization of a novel influenza A reporter virus, which allows fast and sensitive detection of viral spread and its inhibition, and we show that influenza A virus entry is sensitive to alterations of endosomal cholesterol levels.
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Eckert, N., Wrensch, F., Gärtner, S., Palanisamy, N., Goedecke, U., Jäger, N., Pöhlmann, S., & Winkler, M. (2014). Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins. PLoS ONE, 9(5), article-number e97695. https://doi.org/10.1371/journal.pone.0097695
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Public Library of Science
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PLoS ONE
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en
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© 2014 Eckert et al.
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1932-6203
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This work was supported by the Leibniz Graduate School EIDIS (FW) and the Leibniz Foundation.