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Rapid emergence of a PB2-E627K substitution confers a virulent phenotype to an H9N2 avian influenza virus during adaption in mice

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Abstract

The worldwide circulation of H9N2 avian influenza virus in poultry, the greater than 2.3 % positive rate for anti-H9 antibodies in poultry-exposed workers, and several reports of human infection indicate that H9N2 virus is a potential threat to human health. Here, we found three mutations that conferred high virulence to H9N2 virus in mice after four passages. The PB2-E627K substitution rapidly appeared at the second passage and played a decisive role in virulence. Polymerase complexes possessing PB2-E627K displayed 16.1-fold higher viral polymerase activity when compared to the wild-type virus, which may account for enhanced virulence of this virus. The other two substitutions (HA-N313D and HA-N496S) enhanced binding to both α2,3-linked and α2,6-linked sialic acid receptors; however, the HA-N313D and N496S substitutions alone decreased the virulence of mouse-adapted virus. Furthermore, this mouse-adapted virus was still not transmissible among guinea pigs by direct contact (0/3 pairs). Our findings show that adaption in mice enhanced the viral polymerase activity and receptor-binding ability, which resulted in a virulent phenotype in mice but not a transmissible phenotype in guinea pigs, indicating that host factors play an important role in adaptive evolution of influenza in new hosts.

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Acknowledgments

We thank Peter Wilker for editing the manuscript. This work was supported by the Key Projects in the National Science and Technology Pillar Program during the Twelfth Five-Year Plan Period (2012ZX10004-502); National High Technology Research and Development Program (No. 2012AA022006) and the National Science and Technology Major Project (No. 2012ZX1000301008).

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None of the authors report a conflict of interest, and all authors approved the manuscript for publication.

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    1. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150000, Heilongjiang, China

      Xiaoyu Sang, Xijun He & Xiaolong Gao

    2. The Military Veterinary Institute, Academy of Military Medical Science of PLA, Changchun, 130122, People’s Republic of China

      Xiaoyu Sang, Airong Wang, Jie Ding, Xiaolong Gao, Yuanguo Li, Kun Zhang, Zhiguang Ren, Lin Li, Zhijun Yu, Tiecheng Wang, Na Feng, Xuexing Zheng, Hualei Wang, Yongkun Zhao, Songtao Yang, Yuwei Gao & Xianzhu Xia

    3. College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai’an, 271000, Shandong, China

      Airong Wang & Tongjie Chai

    4. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, 130122, Jilin, China

      Yuwei Gao & Xianzhu Xia

    5. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225000, Jiangsu, China

      Yuwei Gao & Xianzhu Xia

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    Sang, X., Wang, A., Chai, T. et al. Rapid emergence of a PB2-E627K substitution confers a virulent phenotype to an H9N2 avian influenza virus during adaption in mice. Arch Virol 160, 1267–1277 (2015). https://doi.org/10.1007/s00705-015-2383-5

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