Viral fusion efficacy of specific H3N2 influenza virus reassortant combinations at single-particle level
Identifieur interne : 000392 ( Main/Exploration ); précédent : 000391; suivant : 000393Viral fusion efficacy of specific H3N2 influenza virus reassortant combinations at single-particle level
Auteurs : Hung-Lun Hsu [États-Unis] ; Jean K. Millet [États-Unis] ; Deirdre A. Costello [États-Unis] ; Gary R. Whittaker [États-Unis] ; Susan Daniel [États-Unis]Source :
- Scientific Reports [ 2045-2322 ] ; 2016.
Abstract
Virus pseudotyping is a useful and safe technique for studying entry of emerging strains of influenza virus. However, few studies have compared different reassortant combinations in pseudoparticle systems, or compared entry kinetics of native viruses and their pseudotyped analogs. Here, vesicular stomatitis virus (VSV)-based pseudovirions displaying distinct influenza virus envelope proteins were tested for fusion activity. We produced VSV pseudotypes containing the prototypical X-31 (H3) HA, either alone or with strain-matched or mismatched N2 NAs. We performed single-particle fusion assays using total internal reflection fluorescence microscopy to compare hemifusion kinetics among these pairings. Results illustrate that matching pseudoparticles behaved very similarly to native virus. Pseudoparticles harboring mismatched HA-NA pairings fuse at significantly slower rates than native virus, and NA-lacking pseudoparticles exhibiting the slowest fusion rates. Relative viral membrane HA density of matching pseudoparticles was higher than in mismatching or NA-lacking pseudoparticles. An equivalent trend of HA expression level on cell membranes of HA/NA co-transfected cells was observed and intracellular trafficking of HA was affected by NA co-expression. Overall, we show that specific influenza HA-NA combinations can profoundly affect the critical role played by HA during entry, which may factor into viral fitness and the emergence of new pandemic influenza viruses.
Url:
DOI: 10.1038/srep35537
PubMed: 27752100
PubMed Central: 5067655
Affiliations:
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Le document en format XML
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<front><div type="abstract" xml:lang="en"><p>Virus pseudotyping is a useful and safe technique for studying entry of emerging strains of influenza virus. However, few studies have compared different reassortant combinations in pseudoparticle systems, or compared entry kinetics of native viruses and their pseudotyped analogs. Here, vesicular stomatitis virus (VSV)-based pseudovirions displaying distinct influenza virus envelope proteins were tested for fusion activity. We produced VSV pseudotypes containing the prototypical X-31 (H3) HA, either alone or with strain-matched or mismatched N2 NAs. We performed single-particle fusion assays using total internal reflection fluorescence microscopy to compare hemifusion kinetics among these pairings. Results illustrate that matching pseudoparticles behaved very similarly to native virus. Pseudoparticles harboring mismatched HA-NA pairings fuse at significantly slower rates than native virus, and NA-lacking pseudoparticles exhibiting the slowest fusion rates. Relative viral membrane HA density of matching pseudoparticles was higher than in mismatching or NA-lacking pseudoparticles. An equivalent trend of HA expression level on cell membranes of HA/NA co-transfected cells was observed and intracellular trafficking of HA was affected by NA co-expression. Overall, we show that specific influenza HA-NA combinations can profoundly affect the critical role played by HA during entry, which may factor into viral fitness and the emergence of new pandemic influenza viruses.</p>
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<author><name sortKey="Daniel, S" uniqKey="Daniel S">S. Daniel</name>
</author>
<author><name sortKey="Cremer, P S" uniqKey="Cremer P">P. S. Cremer</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Costello, D A" uniqKey="Costello D">D. A. Costello</name>
</author>
<author><name sortKey="Millet, J K" uniqKey="Millet J">J. K. Millet</name>
</author>
<author><name sortKey="Hsia, C Y" uniqKey="Hsia C">C.-Y. Hsia</name>
</author>
<author><name sortKey="Whittaker, G R" uniqKey="Whittaker G">G. R. Whittaker</name>
</author>
<author><name sortKey="Daniel, S" uniqKey="Daniel S">S. Daniel</name>
</author>
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<affiliations><list><country><li>États-Unis</li>
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<tree><country name="États-Unis"><noRegion><name sortKey="Hsu, Hung Lun" sort="Hsu, Hung Lun" uniqKey="Hsu H" first="Hung-Lun" last="Hsu">Hung-Lun Hsu</name>
</noRegion>
<name sortKey="Costello, Deirdre A" sort="Costello, Deirdre A" uniqKey="Costello D" first="Deirdre A." last="Costello">Deirdre A. Costello</name>
<name sortKey="Daniel, Susan" sort="Daniel, Susan" uniqKey="Daniel S" first="Susan" last="Daniel">Susan Daniel</name>
<name sortKey="Millet, Jean K" sort="Millet, Jean K" uniqKey="Millet J" first="Jean K." last="Millet">Jean K. Millet</name>
<name sortKey="Whittaker, Gary R" sort="Whittaker, Gary R" uniqKey="Whittaker G" first="Gary R." last="Whittaker">Gary R. Whittaker</name>
</country>
</tree>
</affiliations>
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