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PB2 and HA mutations increase the virulence of highly pathogenic H5N5 clade 2.3.4.4 avian influenza virus in mice

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Abstract

H5 clade 2.3.4.4 influenza A viruses pose a potential threat to public health and are a cause of public concern. Here, we generated mouse-adapted viruses of a waterfowl-origin H5N5 virus (H5 clade 2.3.4.4) to identify adaptive changes that confer increased virulence in mammals. After two passages, we obtained a mouse-adapted H5N5 virus that contained single amino acid substitutions in the PB2 (E627K) and hemagglutinin (HA) (F430L) proteins. We then analyzed the impact of these individual amino acid substitutions on viral pathogenicity to mammals. The 50% mouse lethal dose (MLD50) of the H5N5 virus containing the PB2-E627K substitution or the HA-F430L substitution was reduced 1000-fold or 3.16-fold, respectively. Furthermore, we found that PB2-E627K enhanced viral replication kinetics in vitro and in vivo. These results suggest that the PB2-E627K and HA-F430L substitutions are important for adaptation of H5N5 AIVs to mammals. These findings emphasize the importance of continued surveillance of poultry for H5N5 AIVs with these amino acid substitutions.

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Author notes

  1. Zhijun Yu and Kaihui Cheng contributed equally to the results of this study.

Authors and Affiliations

  1. Institute of Poultry Science, Shandong Academy of Agricultural Sciences, No. 1 Jiaoxiao road, Jinan, 250023, Shandong, China

    Zhijun Yu

  2. Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Military Veterinary Research Institute, Academy of Military Medical Sciences, Changchun, 130122, China

    Weiyang Sun, Xinghai Zhang, Xianzhu Xia & Yuwei Gao

  3. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, China

    Xianzhu Xia & Yuwei Gao

  4. Dairy Cattle Research Center, Shandong Academy of Agricultural Sciences, Jinan, 250132, China

    Kaihui Cheng

  5. The Military Veterinary Institute, Academy of Military Medical Science of PLA, 666 Liuyingxi st, Changchun, 130122, People’s Republic of China

    Xianzhu Xia & Yuwei Gao

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  1. Zhijun Yu
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Correspondence to Zhijun Yu, Xianzhu Xia or Yuwei Gao.

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Funding

This study was funded by the National Key Technology R & D Program (2013BAD12B04), National Key Research and Development Plan (2016YFD0500203), National Key Research and Development Plan (2017YFD0500100), and the High-Level Talents and Innovative Team Recruitment Program of the Shandong Academy of Agricultural Sciences.

Conflict of interest

All authors have read the manuscript and declare that they have no conflicts of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This study does not contain any studies with human participants. The animal studies were conducted within a biosecurity level 3 laboratory approved by the Military Veterinary Research Institute of the Academy of Military Medical Sciences. The protocols for animal studies were approved by the Review Board of Military Veterinary Research Institute of the Academy of Military Medical Sciences.

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Yu, Z., Cheng, K., Sun, W. et al. PB2 and HA mutations increase the virulence of highly pathogenic H5N5 clade 2.3.4.4 avian influenza virus in mice. Arch Virol 163, 401–410 (2018). https://doi.org/10.1007/s00705-017-3631-7

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