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Title: Genetic characterization of an adapted pandemic 2009 H1N1 influenza virus that reveals improved replication rates in human lung epithelial cells

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 (HA{sub 1} D130E, HA{sub 2} 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. - Highlights: • We observed a spontaneous mutation of a 2009-pandemic H1N1 influenza virus in vitro. • The adaptation led to a 100-fold rise in replication rate in human A549 cells. • Adaptation was caused by two mutations in the HA gene segment. • Adaptation correlates with increasedmore » fusion pH and decreased receptor affinity.« less

Authors:
 [1];  [1];  [2]; ;  [1];  [2];  [3];  [1];  [4];  [5]; ;  [4];  [3];  [3];  [2];
  1. Department of Molecular Biology, Max Planck Institute for Infection Biology, Berlin (Germany)
  2. Department of Biology, Molecular Biophysics, IRI Life Sciences, Humboldt-Universität zu Berlin (Germany)
  3. Department for Biomolecular Systems, Max Planck Institute for Colloids and Interfaces, Potsdam (Germany)
  4. Institute for Biophysics, Johannes Kepler University, Linz (Austria)
  5. Center for Advanced Bioanalysis GmbH (CBL), Linz (Austria)
Publication Date:
OSTI Identifier:
22581686
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 492; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AFFINITY; CELL CULTURES; DOMINANT MUTATIONS; GENETICS; HEMAGGLUTININS; IN VITRO; IN VIVO; INFLUENZA; INFLUENZA VIRUSES; LUNGS; MEDICAL SURVEILLANCE; MICE; PH VALUE; RECEPTORS; SPONTANEOUS MUTATIONS; SWINE

Citation Formats

Wörmann, Xenia, Lesch, Markus, Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee, Welke, Robert-William, Okonechnikov, Konstantin, Abdurishid, Mirshat, Sieben, Christian, Geissner, Andreas, Institute of Chemistry and Biochemistry, Free University, Berlin, Brinkmann, Volker, Kastner, Markus, Karner, Andreas, Zhu, Rong, Hinterdorfer, Peter, Anish, Chakkumkal, Seeberger, Peter H., Institute of Chemistry and Biochemistry, Free University, Berlin, Herrmann, Andreas, and others, and. Genetic characterization of an adapted pandemic 2009 H1N1 influenza virus that reveals improved replication rates in human lung epithelial cells. United States: N. p., 2016. Web. doi:10.1016/J.VIROL.2016.02.002.
Wörmann, Xenia, Lesch, Markus, Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee, Welke, Robert-William, Okonechnikov, Konstantin, Abdurishid, Mirshat, Sieben, Christian, Geissner, Andreas, Institute of Chemistry and Biochemistry, Free University, Berlin, Brinkmann, Volker, Kastner, Markus, Karner, Andreas, Zhu, Rong, Hinterdorfer, Peter, Anish, Chakkumkal, Seeberger, Peter H., Institute of Chemistry and Biochemistry, Free University, Berlin, Herrmann, Andreas, & others, and. Genetic characterization of an adapted pandemic 2009 H1N1 influenza virus that reveals improved replication rates in human lung epithelial cells. United States. https://doi.org/10.1016/J.VIROL.2016.02.002
Wörmann, Xenia, Lesch, Markus, Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee, Welke, Robert-William, Okonechnikov, Konstantin, Abdurishid, Mirshat, Sieben, Christian, Geissner, Andreas, Institute of Chemistry and Biochemistry, Free University, Berlin, Brinkmann, Volker, Kastner, Markus, Karner, Andreas, Zhu, Rong, Hinterdorfer, Peter, Anish, Chakkumkal, Seeberger, Peter H., Institute of Chemistry and Biochemistry, Free University, Berlin, Herrmann, Andreas, and others, and. Sun . "Genetic characterization of an adapted pandemic 2009 H1N1 influenza virus that reveals improved replication rates in human lung epithelial cells". United States. https://doi.org/10.1016/J.VIROL.2016.02.002.
@article{osti_22581686,
title = {Genetic characterization of an adapted pandemic 2009 H1N1 influenza virus that reveals improved replication rates in human lung epithelial cells},
author = {Wörmann, Xenia and Lesch, Markus and Steinbeis Innovation gGmbH, Center for Systems Biomedicine, Falkensee and Welke, Robert-William and Okonechnikov, Konstantin and Abdurishid, Mirshat and Sieben, Christian and Geissner, Andreas and Institute of Chemistry and Biochemistry, Free University, Berlin and Brinkmann, Volker and Kastner, Markus and Karner, Andreas and Zhu, Rong and Hinterdorfer, Peter and Anish, Chakkumkal and Seeberger, Peter H. and Institute of Chemistry and Biochemistry, Free University, Berlin and Herrmann, Andreas and others, and},
abstractNote = {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 (HA{sub 1} D130E, HA{sub 2} 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. - Highlights: • We observed a spontaneous mutation of a 2009-pandemic H1N1 influenza virus in vitro. • The adaptation led to a 100-fold rise in replication rate in human A549 cells. • Adaptation was caused by two mutations in the HA gene segment. • Adaptation correlates with increased fusion pH and decreased receptor affinity.},
doi = {10.1016/J.VIROL.2016.02.002},
url = {https://www.osti.gov/biblio/22581686}, journal = {Virology},
issn = {0042-6822},
number = ,
volume = 492,
place = {United States},
year = {2016},
month = {5}
}