Growth Determinants for H5N1 Influenza Vaccine Seed Viruses in MDCK Cells

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Published inJournal of Virology Vol. 82; no. 21; pp. 10502 - 10509
Main Authors Murakami, Shin, Horimoto, Taisuke, Mai, Le Quynh, Nidom, Chairul A., Chen, Hualan, Muramoto, Yukiko, Yamada, Shinya, Iwasa, Ayaka, Iwatsuki-Horimoto, Kiyoko, Shimojima, Masayuki, Iwata, Akira, Kawaoka, Yoshihiro
Format Journal Article
LanguageEnglish
Published Washington, DC American Society for Microbiology 01.11.2008
American Society for Microbiology (ASM)
Subjects
Animals
Biological and medical sciences
Cell Culture Techniques
Cell Line
Dogs
Fundamental and applied biological sciences. Psychology
Influenza A Virus, H5N1 Subtype - genetics
Influenza A Virus, H5N1 Subtype - growth & development
Influenza Vaccines
Microbiology
Miscellaneous
Reassortant Viruses - genetics
Reassortant Viruses - growth & development
Vaccines and Antiviral Agents
Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies
Viral Plaque Assay
Viral Proteins - genetics
Viral Proteins - physiology
Virology
Virus Replication
Infection
Influenzavirus A(H5N1)
Seeds
Viral disease
Influenza
Vaccine
In vitro
Virology
Online AccessGet full text

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Abstract Article Usage Stats Services JVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue JVI About JVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0022-538X Online ISSN: 1098-5514 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JVI .asm.org, visit: JVI       
AbstractList H5N1 influenza A viruses are exacting a growing human toll, with more than 240 fatal cases to date. In the event of an influenza pandemic caused by these viruses, embryonated chicken eggs, which are the approved substrate for human inactivated-vaccine production, will likely be in short supply because chickens will be killed by these viruses or culled to limit the worldwide spread of the infection. The Madin-Darby canine kidney (MDCK) cell line is a promising alternative candidate substrate because it supports efficient growth of influenza viruses compared to other cell lines. Here, we addressed the molecular determinants for growth of an H5N1 vaccine seed virus in MDCK cells, revealing the critical responsibility of the Tyr residue at position 360 of PB2, the considerable requirement for functional balance between hemagglutinin (HA) and neuraminidase (NA), and the partial responsibility of the Glu residue at position 55 of NS1. Based on these findings, we produced a PR8/H5N1 reassortant, optimized for this cell line, that derives all of its genes for its internal proteins from the PR8(UW) strain except for the NS gene, which derives from the PR8(Cambridge) strain; its N1 NA gene, which has a long stalk and derives from an early H5N1 strain; and its HA gene, which has an avirulent-type cleavage site sequence and is derived from a circulating H5N1 virus. Our findings demonstrate the importance and feasibility of a cell culture-based approach to producing seed viruses for inactivated H5N1 vaccines that grow robustly and in a timely, cost-efficient manner as an alternative to egg-based vaccine production.
Article Usage Stats Services JVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue JVI About JVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0022-538X Online ISSN: 1098-5514 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JVI .asm.org, visit: JVI       
H5N1 influenza A viruses are exacting a growing human toll, with more than 240 fatal cases to date. In the event of an influenza pandemic caused by these viruses, embryonated chicken eggs, which are the approved substrate for human inactivated-vaccine production, will likely be in short supply because chickens will be killed by these viruses or culled to limit the worldwide spread of the infection. The Madin-Darby canine kidney (MDCK) cell line is a promising alternative candidate substrate because it supports efficient growth of influenza viruses compared to other cell lines. Here, we addressed the molecular determinants for growth of an H5N1 vaccine seed virus in MDCK cells, revealing the critical responsibility of the Tyr residue at position 360 of PB2, the considerable requirement for functional balance between hemagglutinin (HA) and neuraminidase (NA), and the partial responsibility of the Glu residue at position 55 of NS1. Based on these findings, we produced a PR8/H5N1 reassortant, optimized for this cell line, that derives all of its genes for its internal proteins from the PR8(UW) strain except for the NS gene, which derives from the PR8(Cambridge) strain; its N1 NA gene, which has a long stalk and derives from an early H5N1 strain; and its HA gene, which has an avirulent-type cleavage site sequence and is derived from a circulating H5N1 virus. Our findings demonstrate the importance and feasibility of a cell culture-based approach to producing seed viruses for inactivated H5N1 vaccines that grow robustly and in a timely, cost-efficient manner as an alternative to egg-based vaccine production.H5N1 influenza A viruses are exacting a growing human toll, with more than 240 fatal cases to date. In the event of an influenza pandemic caused by these viruses, embryonated chicken eggs, which are the approved substrate for human inactivated-vaccine production, will likely be in short supply because chickens will be killed by these viruses or culled to limit the worldwide spread of the infection. The Madin-Darby canine kidney (MDCK) cell line is a promising alternative candidate substrate because it supports efficient growth of influenza viruses compared to other cell lines. Here, we addressed the molecular determinants for growth of an H5N1 vaccine seed virus in MDCK cells, revealing the critical responsibility of the Tyr residue at position 360 of PB2, the considerable requirement for functional balance between hemagglutinin (HA) and neuraminidase (NA), and the partial responsibility of the Glu residue at position 55 of NS1. Based on these findings, we produced a PR8/H5N1 reassortant, optimized for this cell line, that derives all of its genes for its internal proteins from the PR8(UW) strain except for the NS gene, which derives from the PR8(Cambridge) strain; its N1 NA gene, which has a long stalk and derives from an early H5N1 strain; and its HA gene, which has an avirulent-type cleavage site sequence and is derived from a circulating H5N1 virus. Our findings demonstrate the importance and feasibility of a cell culture-based approach to producing seed viruses for inactivated H5N1 vaccines that grow robustly and in a timely, cost-efficient manner as an alternative to egg-based vaccine production.
Author Akira Iwata
Kiyoko Iwatsuki-Horimoto
Masayuki Shimojima
Ayaka Iwasa
Yoshihiro Kawaoka
Shinya Yamada
Shin Murakami
Le Quynh Mai
Chairul A. Nidom
Hualan Chen
Taisuke Horimoto
Yukiko Muramoto
AuthorAffiliation Division of Virology, Department of Microbiology and Immunology, 1 International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo, Japan, 2 Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Saitama, Japan, 3 National Institute of Hygiene and Epidemiology, Hanoi, Vietnam, 4 Avian Influenza Laboratory, Tropical Disease Centre, Airlangga University, Surabaya, Indonesia, 5 Animal Influenza Laboratory of the Ministry of Agriculture and National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, People's Republic of China, 6 Nippon Institute for Biological Science, Tokyo, Japan, 7 Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, Wisconsin 8
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  surname: Yamada
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  givenname: Ayaka
  surname: Iwasa
  fullname: Iwasa, Ayaka
  organization: Division of Virology, Department of Microbiology and Immunology, Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Saitama, Japan
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  givenname: Kiyoko
  surname: Iwatsuki-Horimoto
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  surname: Shimojima
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  surname: Iwata
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  surname: Kawaoka
  fullname: Kawaoka, Yoshihiro
  organization: Division of Virology, Department of Microbiology and Immunology, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo, Japan, Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Saitama, Japan, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, Wisconsin
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Issue 21
Keywords Infection
Influenzavirus A(H5N1)
Seeds
Viral disease
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Vaccine
In vitro
Virology
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Corresponding author. Mailing address: Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Phone: 81-3-5449-5281. Fax: 81-3-5449-5408. E-mail for T. Horimoto: horimoto@ims.u-tokyo.ac.jp. E-mail for Y. Kawaoka: kawaoka@ims.u-tokyo.ac.jp
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H5N1 influenza A viruses are exacting a growing human toll, with more than 240 fatal cases to date. In the event of an influenza pandemic caused by these...
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StartPage 10502
SubjectTerms Animals
Biological and medical sciences
Cell Culture Techniques
Cell Line
Dogs
Fundamental and applied biological sciences. Psychology
Influenza A Virus, H5N1 Subtype - genetics
Influenza A Virus, H5N1 Subtype - growth & development
Influenza Vaccines
Microbiology
Miscellaneous
Reassortant Viruses - genetics
Reassortant Viruses - growth & development
Vaccines and Antiviral Agents
Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies
Viral Plaque Assay
Viral Proteins - genetics
Viral Proteins - physiology
Virology
Virus Replication
Title Growth Determinants for H5N1 Influenza Vaccine Seed Viruses in MDCK Cells
URI http://jvi.asm.org/content/82/21/10502.abstract
https://www.ncbi.nlm.nih.gov/pubmed/18768983
https://www.proquest.com/docview/69680291
https://pubmed.ncbi.nlm.nih.gov/PMC2573193
Volume 82
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