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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">A Y161F Hemagglutinin Substitution Increases Thermostability and Improves Yields of 2009 H1N1 Influenza A Virus in Cells</title><author><name sortKey="Wen, Feng" sort="Wen, Feng" uniqKey="Wen F" first="Feng" last="Wen">Feng Wen</name><affiliation><nlm:aff id="aff1">Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author><author><name sortKey="Li, Lei" sort="Li, Lei" uniqKey="Li L" first="Lei" last="Li">Lei Li</name><affiliation><nlm:aff id="aff2">Department of Chemistry, Georgia State University, Atlanta, Georgia, USA</nlm:aff></affiliation></author><author><name sortKey="Zhao, Nan" sort="Zhao, Nan" uniqKey="Zhao N" first="Nan" last="Zhao">Nan Zhao</name><affiliation><nlm:aff id="aff1">Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author><author><name sortKey="Chiang, Meng Jung" sort="Chiang, Meng Jung" uniqKey="Chiang M" first="Meng-Jung" last="Chiang">Meng-Jung Chiang</name><affiliation><nlm:aff id="aff3">Laboratory of Respiratory Viral Diseases, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, United States Food and Drug Administration, Silver Spring, Maryland, USA</nlm:aff></affiliation></author><author><name sortKey="Xie, Hang" sort="Xie, Hang" uniqKey="Xie H" first="Hang" last="Xie">Hang Xie</name><affiliation><nlm:aff id="aff3">Laboratory of Respiratory Viral Diseases, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, United States Food and Drug Administration, Silver Spring, Maryland, USA</nlm:aff></affiliation></author><author><name sortKey="Cooley, Jim" sort="Cooley, Jim" uniqKey="Cooley J" first="Jim" last="Cooley">Jim Cooley</name><affiliation><nlm:aff id="aff4">Department of Population and Pathobiology, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author><author><name sortKey="Webby, Richard" sort="Webby, Richard" uniqKey="Webby R" first="Richard" last="Webby">Richard Webby</name><affiliation><nlm:aff id="aff5">Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee, USA</nlm:aff></affiliation></author><author><name sortKey="Wang, Peng George" sort="Wang, Peng George" uniqKey="Wang P" first="Peng George" last="Wang">Peng George Wang</name><affiliation><nlm:aff id="aff2">Department of Chemistry, Georgia State University, Atlanta, Georgia, USA</nlm:aff></affiliation></author><author><name sortKey="Wan, Xiu Feng" sort="Wan, Xiu Feng" uniqKey="Wan X" first="Xiu-Feng" last="Wan">Xiu-Feng Wan</name><affiliation><nlm:aff id="aff1">Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">29118117</idno><idno type="pmc">5752953</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5752953</idno><idno type="RBID">PMC:5752953</idno><idno type="doi">10.1128/JVI.01621-17</idno><date when="2018">2018</date><idno type="wicri:Area/Pmc/Corpus">000702</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000702</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A Y161F Hemagglutinin Substitution Increases Thermostability and Improves Yields of 2009 H1N1 Influenza A Virus in Cells</title><author><name sortKey="Wen, Feng" sort="Wen, Feng" uniqKey="Wen F" first="Feng" last="Wen">Feng Wen</name><affiliation><nlm:aff id="aff1">Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author><author><name sortKey="Li, Lei" sort="Li, Lei" uniqKey="Li L" first="Lei" last="Li">Lei Li</name><affiliation><nlm:aff id="aff2">Department of Chemistry, Georgia State University, Atlanta, Georgia, USA</nlm:aff></affiliation></author><author><name sortKey="Zhao, Nan" sort="Zhao, Nan" uniqKey="Zhao N" first="Nan" last="Zhao">Nan Zhao</name><affiliation><nlm:aff id="aff1">Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author><author><name sortKey="Chiang, Meng Jung" sort="Chiang, Meng Jung" uniqKey="Chiang M" first="Meng-Jung" last="Chiang">Meng-Jung Chiang</name><affiliation><nlm:aff id="aff3">Laboratory of Respiratory Viral Diseases, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, United States Food and Drug Administration, Silver Spring, Maryland, USA</nlm:aff></affiliation></author><author><name sortKey="Xie, Hang" sort="Xie, Hang" uniqKey="Xie H" first="Hang" last="Xie">Hang Xie</name><affiliation><nlm:aff id="aff3">Laboratory of Respiratory Viral Diseases, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, United States Food and Drug Administration, Silver Spring, Maryland, USA</nlm:aff></affiliation></author><author><name sortKey="Cooley, Jim" sort="Cooley, Jim" uniqKey="Cooley J" first="Jim" last="Cooley">Jim Cooley</name><affiliation><nlm:aff id="aff4">Department of Population and Pathobiology, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author><author><name sortKey="Webby, Richard" sort="Webby, Richard" uniqKey="Webby R" first="Richard" last="Webby">Richard Webby</name><affiliation><nlm:aff id="aff5">Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee, USA</nlm:aff></affiliation></author><author><name sortKey="Wang, Peng George" sort="Wang, Peng George" uniqKey="Wang P" first="Peng George" last="Wang">Peng George Wang</name><affiliation><nlm:aff id="aff2">Department of Chemistry, Georgia State University, Atlanta, Georgia, USA</nlm:aff></affiliation></author><author><name sortKey="Wan, Xiu Feng" sort="Wan, Xiu Feng" uniqKey="Wan X" first="Xiu-Feng" last="Wan">Xiu-Feng Wan</name><affiliation><nlm:aff id="aff1">Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Virology</title><idno type="ISSN">0022-538X</idno><idno type="eISSN">1098-5514</idno><imprint><date when="2018">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>ABSTRACT</title><p>Vaccination is the primary strategy for influenza prevention and control. However, egg-based vaccines, the predominant production platform, have several disadvantages, including the emergence of viral antigenic variants that can be induced during egg passage. These limitations have prompted the development of cell-based vaccines, which themselves are not without issue. Most importantly, vaccine seed viruses often do not grow efficiently in mammalian cell lines. Here we aimed to identify novel high-yield signatures for influenza viruses in continuous Madin-Darby canine kidney (MDCK) and Vero cells. Using influenza A(H1N1)pdm09 virus as the testing platform and an integrating error-prone PCR-based mutagenesis strategy, we identified a Y161F mutation in hemagglutinin (HA) that not only enhanced the infectivity of the resultant virus by more than 300-fold but also increased its thermostability without changing its original antigenic properties. The vaccine produced from the Y161F mutant fully protected mice against lethal challenge with wild-type A(H1N1)pdm09. Compared with A(H1N1)pdm09, the Y161F mutant had significantly higher avidity for avian-like and human-like receptor analogs. Of note, the introduction of the Y161F mutation into HA of seasonal H3N2 influenza A virus (IAV) and canine H3N8 IAV also increased yields and thermostability in MDCK cells and chicken embryotic eggs. Thus, residue F161 plays an important role in determining viral growth and thermostability, which could be harnessed to optimize IAV vaccine seed viruses.</p><p><bold>IMPORTANCE</bold> Although a promising complement to current egg-based influenza vaccines, cell-based vaccines have one large challenge: high-yield vaccine seeds for production. In this study, we identified a molecular signature, Y161F, in hemagglutinin (HA) that resulted in increased virus growth in Madin-Darby canine kidney and Vero cells, two cell lines commonly used for influenza vaccine manufacturing. This Y161F mutation not only increased HA thermostability but also enhanced its binding affinity for α2,6- and α2,3-linked Neu5Ac. These results suggest that a vaccine strain bearing the Y161F mutation in HA could potentially increase vaccine yields in mammalian cell culture systems.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol</journal-id><journal-id journal-id-type="iso-abbrev">J. Virol</journal-id><journal-id journal-id-type="hwp">jvi</journal-id><journal-id journal-id-type="pmc">jvi</journal-id><journal-id journal-id-type="publisher-id">JVI</journal-id><journal-title-group><journal-title>Journal of Virology</journal-title></journal-title-group><issn pub-type="ppub">0022-538X</issn><issn pub-type="epub">1098-5514</issn><publisher><publisher-name>American Society for Microbiology</publisher-name><publisher-loc>1752 N St., N.W., Washington, DC</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">29118117</article-id><article-id pub-id-type="pmc">5752953</article-id><article-id pub-id-type="publisher-id">01621-17</article-id><article-id pub-id-type="doi">10.1128/JVI.01621-17</article-id><article-categories><subj-group subj-group-type="heading"><subject>Vaccines and Antiviral Agents</subject></subj-group></article-categories><title-group><article-title>A Y161F Hemagglutinin Substitution Increases Thermostability and Improves Yields of 2009 H1N1 Influenza A Virus in Cells</article-title><alt-title alt-title-type="running-head">Y161F of HA Increases IAV Yields and Thermostability</alt-title><alt-title alt-title-type="short-authors">Wen et al.</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Wen</surname><given-names>Feng</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Lei</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Zhao</surname><given-names>Nan</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Chiang</surname><given-names>Meng-Jung</given-names></name><xref ref-type="aff" rid="aff3"><sup>c</sup></xref></contrib><contrib contrib-type="author"><name><surname>Xie</surname><given-names>Hang</given-names></name><xref ref-type="aff" rid="aff3"><sup>c</sup></xref></contrib><contrib contrib-type="author"><name><surname>Cooley</surname><given-names>Jim</given-names></name><xref ref-type="aff" rid="aff4"><sup>d</sup></xref></contrib><contrib contrib-type="author"><name><surname>Webby</surname><given-names>Richard</given-names></name><xref ref-type="aff" rid="aff5"><sup>e</sup></xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Peng George</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Wan</surname><given-names>Xiu-Feng</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><aff id="aff1"><label>a</label>Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</aff><aff id="aff2"><label>b</label>Department of Chemistry, Georgia State University, Atlanta, Georgia, USA</aff><aff id="aff3"><label>c</label>Laboratory of Respiratory Viral Diseases, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, United States Food and Drug Administration, Silver Spring, Maryland, USA</aff><aff id="aff4"><label>d</label>Department of Population and Pathobiology, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA</aff><aff id="aff5"><label>e</label>Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee, USA</aff></contrib-group><contrib-group><contrib contrib-type="editor"><name><surname>Sandri-Goldin</surname><given-names>Rozanne M.</given-names></name><role>Editor</role><aff>University of California, Irvine</aff></contrib></contrib-group><author-notes><corresp id="cor1">Address correspondence to Xiu-Feng Wan, <email>wan@cvm.msstate.edu</email>.</corresp><fn fn-type="other"><p><bold>Citation</bold> Wen F, Li L, Zhao N, Chiang M-J, Xie H, Cooley J, Webby R, Wang PG, Wan X-F. 2018. A Y161F hemagglutinin substitution increases thermostability and improves yields of 2009 H1N1 influenza A virus in cells. J Virol 92:e01621-17. <ext-link ext-link-type="uri" xlink:href="https://doi.org/10.1128/JVI.01621-17">https://doi.org/10.1128/JVI.01621-17</ext-link>.</p></fn></author-notes><pub-date pub-type="epreprint"><day>8</day><month>11</month><year>2017</year></pub-date><pub-date pub-type="epub"><day>2</day><month>1</month><year>2018</year></pub-date><pub-date pub-type="collection"><day>15</day><month>1</month><year>2018</year></pub-date><volume>92</volume><issue>2</issue><elocation-id>e01621-17</elocation-id><history><date date-type="received"><day>12</day><month>9</month><year>2017</year></date><date date-type="accepted"><day>27</day><month>10</month><year>2017</year></date></history><permissions><copyright-statement>Copyright © 2018 American Society for Microbiology.</copyright-statement><copyright-year>2018</copyright-year><copyright-holder>American Society for Microbiology</copyright-holder><license license-type="asm" xlink:href="https://doi.org/10.1128/ASMCopyrightv2"><license-p><ext-link ext-link-type="uri" xlink:href="https://doi.org/10.1128/ASMCopyrightv2">All Rights Reserved</ext-link>.</license-p></license></permissions><self-uri content-type="pdf" xlink:href="zjv002183246001.pdf"></self-uri><abstract><title>ABSTRACT</title><p>Vaccination is the primary strategy for influenza prevention and control. However, egg-based vaccines, the predominant production platform, have several disadvantages, including the emergence of viral antigenic variants that can be induced during egg passage. These limitations have prompted the development of cell-based vaccines, which themselves are not without issue. Most importantly, vaccine seed viruses often do not grow efficiently in mammalian cell lines. Here we aimed to identify novel high-yield signatures for influenza viruses in continuous Madin-Darby canine kidney (MDCK) and Vero cells. Using influenza A(H1N1)pdm09 virus as the testing platform and an integrating error-prone PCR-based mutagenesis strategy, we identified a Y161F mutation in hemagglutinin (HA) that not only enhanced the infectivity of the resultant virus by more than 300-fold but also increased its thermostability without changing its original antigenic properties. The vaccine produced from the Y161F mutant fully protected mice against lethal challenge with wild-type A(H1N1)pdm09. Compared with A(H1N1)pdm09, the Y161F mutant had significantly higher avidity for avian-like and human-like receptor analogs. Of note, the introduction of the Y161F mutation into HA of seasonal H3N2 influenza A virus (IAV) and canine H3N8 IAV also increased yields and thermostability in MDCK cells and chicken embryotic eggs. Thus, residue F161 plays an important role in determining viral growth and thermostability, which could be harnessed to optimize IAV vaccine seed viruses.</p><p><bold>IMPORTANCE</bold> Although a promising complement to current egg-based influenza vaccines, cell-based vaccines have one large challenge: high-yield vaccine seeds for production. In this study, we identified a molecular signature, Y161F, in hemagglutinin (HA) that resulted in increased virus growth in Madin-Darby canine kidney and Vero cells, two cell lines commonly used for influenza vaccine manufacturing. This Y161F mutation not only increased HA thermostability but also enhanced its binding affinity for α2,6- and α2,3-linked Neu5Ac. These results suggest that a vaccine strain bearing the Y161F mutation in HA could potentially increase vaccine yields in mammalian cell culture systems.</p></abstract><kwd-group><title>KEYWORDS</title><kwd>2009 H1N1 virus</kwd><kwd>H3N2</kwd><kwd>H3N8</kwd><kwd>MDCK</kwd><kwd>Vero cells</kwd><kwd>Y161F</kwd><kwd>high yields</kwd><kwd>influenza A virus</kwd><kwd>vaccines</kwd></kwd-group><funding-group><award-group id="award1"><funding-source id="gs1"><institution-wrap><institution>HHS | National Institutes of Health (NIH)</institution><institution-id>https://doi.org/10.13039/100000002</institution-id></institution-wrap></funding-source><award-id rid="gs1">P20GM103646</award-id><principal-award-recipient><name><surname>Wan</surname><given-names>Xiu-Feng</given-names></name></principal-award-recipient></award-group><award-group id="award2"><funding-source id="gs2"><institution-wrap><institution>HHS | NIH | National Institute of Allergy and Infectious Diseases (NIAID)</institution><institution-id>https://doi.org/10.13039/100000060</institution-id></institution-wrap></funding-source><award-id rid="gs2">R01AI116744</award-id><principal-award-recipient><name><surname>Wan</surname><given-names>Xiu-Feng</given-names></name></principal-award-recipient></award-group></funding-group><counts><count count="1" count-type="supplementary-material"></count><fig-count count="6"></fig-count><table-count count="3"></table-count><equation-count count="0"></equation-count><ref-count count="53"></ref-count><page-count count="17"></page-count><word-count count="10328"></word-count></counts><custom-meta-group><custom-meta><meta-name>cover-date</meta-name><meta-value>January 2018</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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