Genetic characterization of an adapted pandemic 2009 H1N1 influenza virus that reveals improved replication rates in human lung epithelial cells.
Identifieur interne : 000884 ( Main/Exploration ); précédent : 000883; suivant : 000885Genetic characterization of an adapted pandemic 2009 H1N1 influenza virus that reveals improved replication rates in human lung epithelial cells.
Auteurs : Xenia Wörmann [Allemagne] ; Markus Lesch [Allemagne] ; Robert-William Welke [Allemagne] ; Konstantin Okonechnikov [Allemagne] ; Mirshat Abdurishid [Allemagne] ; Christian Sieben [Allemagne] ; Andreas Geissner [Allemagne] ; Volker Brinkmann [Allemagne] ; Markus Kastner [Autriche] ; Andreas Karner [Autriche] ; Rong Zhu [Autriche] ; Peter Hinterdorfer [Autriche] ; Chakkumkal Anish [Allemagne] ; Peter H. Seeberger [Allemagne] ; Andreas Herrmann [Allemagne] ; Thomas F. Meyer [Allemagne] ; Alexander Karlas [Allemagne]Source :
- Virology [ 1096-0341 ] ; 2016.
Descripteurs français
- KwdFr :
- ARN viral (génétique), ARN viral (métabolisme), Adaptation physiologique (génétique), Animaux, Cellules rénales canines Madin-Darby, Cellules épithéliales (virologie), Chiens, Expression des gènes, Glycoprotéine hémagglutinine du virus influenza (), Glycoprotéine hémagglutinine du virus influenza (génétique), Glycoprotéine hémagglutinine du virus influenza (métabolisme), Génétique inverse, Humains, Interactions hôte-pathogène, Modèles moléculaires, Passage en série, Poulets, Récepteurs viraux (génétique), Récepteurs viraux (métabolisme), Réplication virale (génétique), Souris, Souris de lignée BALB C, Sous-type H1N1 du virus de la grippe A (croissance et développement), Sous-type H1N1 du virus de la grippe A (génétique), Structure secondaire des protéines, Structure tertiaire des protéines, Séquençage nucléotidique à haut débit.
- MESH :
- croissance et développement : Sous-type H1N1 du virus de la grippe A.
- génétique : ARN viral, Adaptation physiologique, Glycoprotéine hémagglutinine du virus influenza, Récepteurs viraux, Réplication virale, Sous-type H1N1 du virus de la grippe A.
- métabolisme : ARN viral, Glycoprotéine hémagglutinine du virus influenza, Récepteurs viraux.
- virologie : Cellules épithéliales.
- Animaux, Cellules rénales canines Madin-Darby, Chiens, Expression des gènes, Glycoprotéine hémagglutinine du virus influenza, Génétique inverse, Humains, Interactions hôte-pathogène, Modèles moléculaires, Passage en série, Poulets, Souris, Souris de lignée BALB C, Structure secondaire des protéines, Structure tertiaire des protéines, Séquençage nucléotidique à haut débit.
English descriptors
- KwdEn :
- Adaptation, Physiological (genetics), Animals, Chickens, Dogs, Epithelial Cells (virology), Gene Expression, Hemagglutinin Glycoproteins, Influenza Virus (chemistry), Hemagglutinin Glycoproteins, Influenza Virus (genetics), Hemagglutinin Glycoproteins, Influenza Virus (metabolism), High-Throughput Nucleotide Sequencing, Host-Pathogen Interactions, Humans, Influenza A Virus, H1N1 Subtype (genetics), Influenza A Virus, H1N1 Subtype (growth & development), Madin Darby Canine Kidney Cells, Mice, Mice, Inbred BALB C, Models, Molecular, Protein Structure, Secondary, Protein Structure, Tertiary, RNA, Viral (genetics), RNA, Viral (metabolism), Receptors, Virus (genetics), Receptors, Virus (metabolism), Reverse Genetics, Serial Passage, Virus Replication (genetics).
- MESH :
- chemical , chemistry : Hemagglutinin Glycoproteins, Influenza Virus.
- genetics : Adaptation, Physiological, Hemagglutinin Glycoproteins, Influenza Virus, Influenza A Virus, H1N1 Subtype, RNA, Viral, Receptors, Virus, Virus Replication.
- growth & development : Influenza A Virus, H1N1 Subtype.
- chemical , metabolism : Hemagglutinin Glycoproteins, Influenza Virus, RNA, Viral, Receptors, Virus.
- virology : Epithelial Cells.
- Animals, Chickens, Dogs, Gene Expression, High-Throughput Nucleotide Sequencing, Host-Pathogen Interactions, Humans, Madin Darby Canine Kidney Cells, Mice, Mice, Inbred BALB C, Models, Molecular, Protein Structure, Secondary, Protein Structure, Tertiary, Reverse Genetics, Serial Passage.
Abstract
The 2009 influenza pandemic originated from a swine-origin H1N1 virus, which, although less pathogenic than anticipated, may acquire additional virulence-associated mutations in the future. To estimate the potential risk, we sequentially passaged the isolate A/Hamburg/04/2009 in A549 human lung epithelial cells. After passage 6, we observed a 100-fold increased replication rate. High-throughput sequencing of viral gene segments identified five dominant mutations, whose contribution to the enhanced growth was analyzed by reverse genetics. The increased replication rate was pinpointed to two mutations within the hemagglutinin (HA) gene segment (HA1 D130E, HA2 I91L), near the receptor binding site and the stem domain. The adapted virus also replicated more efficiently in mice in vivo. Enhanced replication rate correlated with increased fusion pH of the HA protein and a decrease in receptor affinity. Our data might be relevant for surveillance of pre-pandemic strains and development of high titer cell culture strains for vaccine production.
DOI: 10.1016/j.virol.2016.02.002
PubMed: 26914510
Affiliations:
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Le document en format XML
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<affiliation wicri:level="1"><nlm:affiliation>Institute for Biophysics, Johannes Kepler University, Linz, Austria.</nlm:affiliation>
<country xml:lang="fr">Autriche</country>
<wicri:regionArea>Institute for Biophysics, Johannes Kepler University, Linz</wicri:regionArea>
<wicri:noRegion>Linz</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Anish, Chakkumkal" sort="Anish, Chakkumkal" uniqKey="Anish C" first="Chakkumkal" last="Anish">Chakkumkal Anish</name>
<affiliation wicri:level="3"><nlm:affiliation>Department for Biomolecular Systems, Max Planck Institute for Colloids and Interfaces, Potsdam, Germany.</nlm:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Department for Biomolecular Systems, Max Planck Institute for Colloids and Interfaces, Potsdam</wicri:regionArea>
<placeName><region type="land" nuts="2">Brandebourg</region>
<settlement type="city">Potsdam</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="Seeberger, Peter H" sort="Seeberger, Peter H" uniqKey="Seeberger P" first="Peter H" last="Seeberger">Peter H. Seeberger</name>
<affiliation wicri:level="3"><nlm:affiliation>Department for Biomolecular Systems, Max Planck Institute for Colloids and Interfaces, Potsdam, Germany; Institute of Chemistry and Biochemistry, Free University, Berlin, Germany.</nlm:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Department for Biomolecular Systems, Max Planck Institute for Colloids and Interfaces, Potsdam, Germany; Institute of Chemistry and Biochemistry, Free University, Berlin</wicri:regionArea>
<placeName><region type="land" nuts="3">Berlin</region>
<settlement type="city">Berlin</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="Herrmann, Andreas" sort="Herrmann, Andreas" uniqKey="Herrmann A" first="Andreas" last="Herrmann">Andreas Herrmann</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Biology, Molecular Biophysics, IRI Life Sciences, Humboldt-Universität zu Berlin, Germany.</nlm:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Department of Biology, Molecular Biophysics, IRI Life Sciences, Humboldt-Universität zu Berlin</wicri:regionArea>
<wicri:noRegion>Humboldt-Universität zu Berlin</wicri:noRegion>
<wicri:noRegion>Humboldt-Universität zu Berlin</wicri:noRegion>
<wicri:noRegion>Humboldt-Universität zu Berlin</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Meyer, Thomas F" sort="Meyer, Thomas F" uniqKey="Meyer T" first="Thomas F" last="Meyer">Thomas F. Meyer</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Molecular Biology, Max Planck Institute for Infection Biology, Berlin, Germany; Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee, Germany. Electronic address: meyer@mpiib-berlin.mpg.de.</nlm:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Department of Molecular Biology, Max Planck Institute for Infection Biology, Berlin, Germany; Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee</wicri:regionArea>
<wicri:noRegion>Falkensee</wicri:noRegion>
<wicri:noRegion>Falkensee</wicri:noRegion>
<wicri:noRegion>Falkensee</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Karlas, Alexander" sort="Karlas, Alexander" uniqKey="Karlas A" first="Alexander" last="Karlas">Alexander Karlas</name>
<affiliation wicri:level="1"><nlm:affiliation>Department of Molecular Biology, Max Planck Institute for Infection Biology, Berlin, Germany; Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee, Germany. Electronic address: karlas@mpiib-berlin.mpg.de.</nlm:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Department of Molecular Biology, Max Planck Institute for Infection Biology, Berlin, Germany; Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee</wicri:regionArea>
<wicri:noRegion>Falkensee</wicri:noRegion>
<wicri:noRegion>Falkensee</wicri:noRegion>
<wicri:noRegion>Falkensee</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j">Virology</title>
<idno type="eISSN">1096-0341</idno>
<imprint><date when="2016" type="published">2016</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adaptation, Physiological (genetics)</term>
<term>Animals</term>
<term>Chickens</term>
<term>Dogs</term>
<term>Epithelial Cells (virology)</term>
<term>Gene Expression</term>
<term>Hemagglutinin Glycoproteins, Influenza Virus (chemistry)</term>
<term>Hemagglutinin Glycoproteins, Influenza Virus (genetics)</term>
<term>Hemagglutinin Glycoproteins, Influenza Virus (metabolism)</term>
<term>High-Throughput Nucleotide Sequencing</term>
<term>Host-Pathogen Interactions</term>
<term>Humans</term>
<term>Influenza A Virus, H1N1 Subtype (genetics)</term>
<term>Influenza A Virus, H1N1 Subtype (growth & development)</term>
<term>Madin Darby Canine Kidney Cells</term>
<term>Mice</term>
<term>Mice, Inbred BALB C</term>
<term>Models, Molecular</term>
<term>Protein Structure, Secondary</term>
<term>Protein Structure, Tertiary</term>
<term>RNA, Viral (genetics)</term>
<term>RNA, Viral (metabolism)</term>
<term>Receptors, Virus (genetics)</term>
<term>Receptors, Virus (metabolism)</term>
<term>Reverse Genetics</term>
<term>Serial Passage</term>
<term>Virus Replication (genetics)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (génétique)</term>
<term>ARN viral (métabolisme)</term>
<term>Adaptation physiologique (génétique)</term>
<term>Animaux</term>
<term>Cellules rénales canines Madin-Darby</term>
<term>Cellules épithéliales (virologie)</term>
<term>Chiens</term>
<term>Expression des gènes</term>
<term>Glycoprotéine hémagglutinine du virus influenza ()</term>
<term>Glycoprotéine hémagglutinine du virus influenza (génétique)</term>
<term>Glycoprotéine hémagglutinine du virus influenza (métabolisme)</term>
<term>Génétique inverse</term>
<term>Humains</term>
<term>Interactions hôte-pathogène</term>
<term>Modèles moléculaires</term>
<term>Passage en série</term>
<term>Poulets</term>
<term>Récepteurs viraux (génétique)</term>
<term>Récepteurs viraux (métabolisme)</term>
<term>Réplication virale (génétique)</term>
<term>Souris</term>
<term>Souris de lignée BALB C</term>
<term>Sous-type H1N1 du virus de la grippe A (croissance et développement)</term>
<term>Sous-type H1N1 du virus de la grippe A (génétique)</term>
<term>Structure secondaire des protéines</term>
<term>Structure tertiaire des protéines</term>
<term>Séquençage nucléotidique à haut débit</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Hemagglutinin Glycoproteins, Influenza Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Sous-type H1N1 du virus de la grippe A</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Adaptation, Physiological</term>
<term>Hemagglutinin Glycoproteins, Influenza Virus</term>
<term>Influenza A Virus, H1N1 Subtype</term>
<term>RNA, Viral</term>
<term>Receptors, Virus</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Influenza A Virus, H1N1 Subtype</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term>
<term>Adaptation physiologique</term>
<term>Glycoprotéine hémagglutinine du virus influenza</term>
<term>Récepteurs viraux</term>
<term>Réplication virale</term>
<term>Sous-type H1N1 du virus de la grippe A</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Hemagglutinin Glycoproteins, Influenza Virus</term>
<term>RNA, Viral</term>
<term>Receptors, Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN viral</term>
<term>Glycoprotéine hémagglutinine du virus influenza</term>
<term>Récepteurs viraux</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Cellules épithéliales</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Epithelial Cells</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Chickens</term>
<term>Dogs</term>
<term>Gene Expression</term>
<term>High-Throughput Nucleotide Sequencing</term>
<term>Host-Pathogen Interactions</term>
<term>Humans</term>
<term>Madin Darby Canine Kidney Cells</term>
<term>Mice</term>
<term>Mice, Inbred BALB C</term>
<term>Models, Molecular</term>
<term>Protein Structure, Secondary</term>
<term>Protein Structure, Tertiary</term>
<term>Reverse Genetics</term>
<term>Serial Passage</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Cellules rénales canines Madin-Darby</term>
<term>Chiens</term>
<term>Expression des gènes</term>
<term>Glycoprotéine hémagglutinine du virus influenza</term>
<term>Génétique inverse</term>
<term>Humains</term>
<term>Interactions hôte-pathogène</term>
<term>Modèles moléculaires</term>
<term>Passage en série</term>
<term>Poulets</term>
<term>Souris</term>
<term>Souris de lignée BALB C</term>
<term>Structure secondaire des protéines</term>
<term>Structure tertiaire des protéines</term>
<term>Séquençage nucléotidique à haut débit</term>
</keywords>
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<front><div type="abstract" xml:lang="en">The 2009 influenza pandemic originated from a swine-origin H1N1 virus, which, although less pathogenic than anticipated, may acquire additional virulence-associated mutations in the future. To estimate the potential risk, we sequentially passaged the isolate A/Hamburg/04/2009 in A549 human lung epithelial cells. After passage 6, we observed a 100-fold increased replication rate. High-throughput sequencing of viral gene segments identified five dominant mutations, whose contribution to the enhanced growth was analyzed by reverse genetics. The increased replication rate was pinpointed to two mutations within the hemagglutinin (HA) gene segment (HA1 D130E, HA2 I91L), near the receptor binding site and the stem domain. The adapted virus also replicated more efficiently in mice in vivo. Enhanced replication rate correlated with increased fusion pH of the HA protein and a decrease in receptor affinity. Our data might be relevant for surveillance of pre-pandemic strains and development of high titer cell culture strains for vaccine production. </div>
</front>
</TEI>
<affiliations><list><country><li>Allemagne</li>
<li>Autriche</li>
</country>
<region><li>Berlin</li>
<li>Brandebourg</li>
</region>
<settlement><li>Berlin</li>
<li>Potsdam</li>
</settlement>
</list>
<tree><country name="Allemagne"><region name="Berlin"><name sortKey="Wormann, Xenia" sort="Wormann, Xenia" uniqKey="Wormann X" first="Xenia" last="Wörmann">Xenia Wörmann</name>
</region>
<name sortKey="Abdurishid, Mirshat" sort="Abdurishid, Mirshat" uniqKey="Abdurishid M" first="Mirshat" last="Abdurishid">Mirshat Abdurishid</name>
<name sortKey="Anish, Chakkumkal" sort="Anish, Chakkumkal" uniqKey="Anish C" first="Chakkumkal" last="Anish">Chakkumkal Anish</name>
<name sortKey="Brinkmann, Volker" sort="Brinkmann, Volker" uniqKey="Brinkmann V" first="Volker" last="Brinkmann">Volker Brinkmann</name>
<name sortKey="Geissner, Andreas" sort="Geissner, Andreas" uniqKey="Geissner A" first="Andreas" last="Geissner">Andreas Geissner</name>
<name sortKey="Herrmann, Andreas" sort="Herrmann, Andreas" uniqKey="Herrmann A" first="Andreas" last="Herrmann">Andreas Herrmann</name>
<name sortKey="Karlas, Alexander" sort="Karlas, Alexander" uniqKey="Karlas A" first="Alexander" last="Karlas">Alexander Karlas</name>
<name sortKey="Lesch, Markus" sort="Lesch, Markus" uniqKey="Lesch M" first="Markus" last="Lesch">Markus Lesch</name>
<name sortKey="Meyer, Thomas F" sort="Meyer, Thomas F" uniqKey="Meyer T" first="Thomas F" last="Meyer">Thomas F. Meyer</name>
<name sortKey="Okonechnikov, Konstantin" sort="Okonechnikov, Konstantin" uniqKey="Okonechnikov K" first="Konstantin" last="Okonechnikov">Konstantin Okonechnikov</name>
<name sortKey="Seeberger, Peter H" sort="Seeberger, Peter H" uniqKey="Seeberger P" first="Peter H" last="Seeberger">Peter H. Seeberger</name>
<name sortKey="Sieben, Christian" sort="Sieben, Christian" uniqKey="Sieben C" first="Christian" last="Sieben">Christian Sieben</name>
<name sortKey="Welke, Robert William" sort="Welke, Robert William" uniqKey="Welke R" first="Robert-William" last="Welke">Robert-William Welke</name>
</country>
<country name="Autriche"><noRegion><name sortKey="Kastner, Markus" sort="Kastner, Markus" uniqKey="Kastner M" first="Markus" last="Kastner">Markus Kastner</name>
</noRegion>
<name sortKey="Hinterdorfer, Peter" sort="Hinterdorfer, Peter" uniqKey="Hinterdorfer P" first="Peter" last="Hinterdorfer">Peter Hinterdorfer</name>
<name sortKey="Karner, Andreas" sort="Karner, Andreas" uniqKey="Karner A" first="Andreas" last="Karner">Andreas Karner</name>
<name sortKey="Zhu, Rong" sort="Zhu, Rong" uniqKey="Zhu R" first="Rong" last="Zhu">Rong Zhu</name>
</country>
</tree>
</affiliations>
</record>
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