Jpn. J. Infect. Dis., 65 (1), 19-27, 2012
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Kazuo Ohnishi1, Yoshimasa Takahashi1, Naoko Kono2, Noriko Nakajima3, Fuminori Mizukoshi1, Shuhei Misawa4, Takuya Yamamoto1, Yu-ya Mitsuki1, Shu-ichi Fu1, Nakami Hirayama1, Masamichi Ohshima1, Manabu Ato1, Tsutomu Kageyama2, Takato Odagiri2, Masato Tashiro2, Kazuo Kobayashi1, Shigeyuki Itamura2, and Yasuko Tsunetsugu-Yokota1*
1Department of Immunology, 2Influenza Virus Research Center, and 3Department of Pathology, National Institute of Infectious Diseases, Tokyo 162-8640; and 4Tsuruga Institute of Biotechnology, Toyobo, Co., Ltd., Fukui 914-8550, Japan
(Received October 4, 2011. Accepted October 28, 2011)
*Corresponding author: Mailing address: Department of Immunology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan. Tel: +81-3-5285-1111, Fax: +81-5285-1150, E-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.
SUMMARY: 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.