Newly Established Monoclonal Antibodies for Immunological Detection of H5N1 Influenza Virus

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  • Ohnishi Kazuo
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Takahashi Yoshimasa
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Kono Naoko
    Influenza Virus Research Center, National Institute of Infectious Diseases, Japan
  • Nakajima Noriko
    Department of Pathology, National Institute of Infectious Diseases, Japan
  • Mizukoshi Fuminori
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Misawa Shuhei
    Tsuruga Institute of Biotechnology, Toyobo, Co., Ltd., Japan
  • Yamamoto Takuya
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Mitsuki Yu-ya
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Fu Shu-ichi
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Hirayama Nakami
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Ohshima Masamichi
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Ato Manabu
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Kageyama Tsutomu
    Influenza Virus Research Center, National Institute of Infectious Diseases, Japan
  • Odagiri Takato
    Influenza Virus Research Center, National Institute of Infectious Diseases, Japan
  • Tashiro Masato
    Influenza Virus Research Center, National Institute of Infectious Diseases, Japan
  • Kobayashi Kazuo
    Department of Immunology, National Institute of Infectious Diseases, Japan
  • Itamura Shigeyuki
    Influenza Virus Research Center, National Institute of Infectious Diseases, Japan
  • Tsunetsugu-Yokota Yasuko
    Department of Immunology, National Institute of Infectious Diseases, Japan

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Abstract

<p>The H5N1 subtype of the highly pathogenic (HP) avian influenza virus has been recognized for its ability to cause serious pandemics among humans. In the present study, new monoclonal antibodies (mAbs) against viral proteins were established for the immunological detection of H5N1 influenza virus for research and diagnostic purposes. B-cell hybridomas were generated from mice that had been hyperimmunized with purified A/Vietnam/1194/2004 (NIBRG-14) virion that had been inactivated by UV-irradiation or formaldehyde. After screening over 4,000 hybridomas, eight H5N1-specific clones were selected. Six were specific for hemagglutinin (HA) and had in vitro neutralization activity. Of these, four were able to broadly detect all tested clades of the H5N1 strains. Five HA-specific mAbs detected denatured HA epitope(s) in Western blot analysis, and two detected HP influenza virus by immunofluorescence and immunohistochemistry. A highly sensitive antigen-capture sandwich ELISA system was established by combining mAbs with different specificities. In conclusion, these mAbs may be useful for rapid and specific diagnosis of H5N1 influenza. Therapeutically, they may have a role in antibody-based treatment of the disease.</p>

Journal

  • Japanese Journal of Infectious Diseases

    Japanese Journal of Infectious Diseases 65 (1), 19-27, 2012-01-31

    National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee

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