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Development of a dual-protective live attenuated vaccine against H5N1 and H9N2 avian influenza viruses by modifying the NS1 gene

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Abstract

An increasing number of outbreaks of avian influenza H5N1 and H9N2 viruses in poultry have caused serious economic losses and raised concerns for human health due to the risk of zoonotic transmission. However, licensed H5N1 and H9N2 vaccines for animals and humans have not been developed. Thus, to develop a dual H5N1 and H9N2 live-attenuated influenza vaccine (LAIV), the HA and NA genes from a virulent mouse-adapted avian H5N2 (A/WB/Korea/ma81/06) virus and a recently isolated chicken H9N2 (A/CK/Korea/116/06) virus, respectively, were introduced into the A/Puerto Rico/8/34 backbone expressing truncated NS1 proteins (NS1-73, NS1-86, NS1-101, NS1-122) but still possessing a full-length NS gene. Two H5N2/NS1-LAIV viruses (H5N2/NS1-86 and H5N2/NS1-101) were highly attenuated compared with the full-length and remaining H5N2/NS-LAIV viruses in a mouse model. Furthermore, viruses containing NS1 modifications were found to induce more IFN-β activation than viruses with full-length NS1 proteins and were correspondingly attenuated in mice. Intranasal vaccination with a single dose (104.0 PFU/ml) of these viruses completely protected mice from a lethal challenge with the homologous A/WB/Korea/ma81/06 (H5N2), heterologous highly pathogenic A/EM/Korea/W149/06 (H5N1), and heterosubtypic highly virulent mouse-adapted H9N2 viruses. This study clearly demonstrates that the modified H5N2/NS1-LAIV viruses attenuated through the introduction of mutations in the NS1 coding region display characteristics that are desirable for live attenuated vaccines and hold potential as vaccine candidates for mammalian hosts.

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Acknowledgments

This work was supported by a National Agenda Project grant from the Korea Research Council of Fundamental Science & Technology and the KRIBB Initiative Program (KGM3111013), and by Korea Healthcare Technology R&D Project funded by the Ministry of Health (Grant No: A103001).

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    Authors and Affiliations

    1. Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University, 12 Gaeshin-Dong Heungduk-Ku, Cheongju, 361-763, Republic of Korea

      Eun-hye Choi, Min-Suk Song, Su-Jin Park, Philippe Noriel Q. Pascua, Yun Hee Baek, Hyeok-il Kwon, Eun-Ha Kim, Semi Kim & Young Ki Choi

    2. Departments of Infectious Diseases and Avian Diseases, College of Veterinary Medicine and Korea Zoonosis Research Institute, Chonbuk National University, Chonbuk, Republic of Korea

      Hyung-Kwan Jang

    3. Korean Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea

      Haryoung Poo

    4. College of Veterinary Medicine, Chungnam National University, 220 Gung-Dong, Yuseoung-Gu, Daejeon, 305-764, Republic of Korea

      Chul-Joong Kim

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    1. Eun-hye Choi
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    Correspondence to Young Ki Choi.

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    E. Choi, M.-S. Song and S.-J. Park contributed equally to this work.

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    Choi, Eh., Song, MS., Park, SJ. et al. Development of a dual-protective live attenuated vaccine against H5N1 and H9N2 avian influenza viruses by modifying the NS1 gene. Arch Virol 160, 1729–1740 (2015). https://doi.org/10.1007/s00705-015-2442-y

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