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Attenuation of an influenza A virus due to alteration of its hemagglutinin-neuraminidase functional balance in mice

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Abstract

Influenza A viruses possess two surface glycoproteins, hemagglutinin (HA), which binds to sialic-acid-containing receptors, and neuraminidase (NA), which removes sialic acid from host cells. It is well established that the HA-NA functional balance regulates the efficiency of virus replication. Here, we selected a plaque variant of the WSN (H1N1) strain that grew better than the wild-type virus in NA-expressing MDCK cell culture. A reverse genetics study revealed that the single mutation HA E190K, which occurs infrequently in naturally isolated H1N1 viruses, was responsible for the phenotype of this variant. Receptor assays indicated that this mutation did not affect the receptor specificity of HA but enhanced its receptor-binding affinity, resulting in altered HA-NA functional balance relative to that of the wild-type virus. We also found that this variant replicated in nasal turbinates at an equivalent level but in lungs at a lower level compared with wild-type virus, demonstrating its attenuation in mice. Together, our data demonstrated the importance of the HA-NA functional balance for influenza virus replication in an in vivo biological setting.

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Acknowledgments

We thank Dr. Emi Takashita and Susan Watson for providing anti-NA antibody and editing the manuscript. This work was supported by grants-in-aid for Specially Promoted Research and for Scientific Research (B), from the Ministry of Education, Culture, Sports, Science, and Technology, by grants-in-aid from the Ministry of Health, Labor and Welfare, by ERATO, and by National Institute of Allergy and Infectious Diseases Public Health Service research grants.

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The authors declare that they have no conflicts of interest

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Authors and Affiliations

  1. Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan

    Fumihiro Gen, Kentaro Kato, Hiroomi Akashi & Taisuke Horimoto

  2. Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, The University of Tokyo, Tokyo, 108-8639, Japan

    Shinya Yamada & Yoshihiro Kawaoka

  3. ERATO Infection-Induced Host Responses Project, Saitama, 332-0012, Japan

    Yoshihiro Kawaoka

  4. Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Yoshihiro Kawaoka

  5. International Research Center for Infectious Diseases, Institute of Medical Science, The University of Tokyo, Tokyo, 108-8639, Japan

    Yoshihiro Kawaoka

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  1. Fumihiro Gen
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  2. Shinya Yamada
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  3. Kentaro Kato
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  4. Hiroomi Akashi
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  5. Yoshihiro Kawaoka
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  6. Taisuke Horimoto
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Correspondence to Taisuke Horimoto.

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Gen, F., Yamada, S., Kato, K. et al. Attenuation of an influenza A virus due to alteration of its hemagglutinin-neuraminidase functional balance in mice. Arch Virol 158, 1003–1011 (2013). https://doi.org/10.1007/s00705-012-1577-3

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  • DOI: https://doi.org/10.1007/s00705-012-1577-3

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