Molecular characterization of mammalian-adapted Korean-type avian H9N2 virus and evaluation of its virulence in mice

Kuk Jin Park; Min-Suk Song; Eun-Ha Kim; Hyeok-il Kwon; Yun Hee Baek; Eun-hye Choi; Su-Jin Park; Se Mi Kim; Young-il Kim; Won-Suk Choi; Dae-Won Yoo; Chul-Joong Kim; Young Ki Choi

doi:10.1007/s12275-015-5329-4

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Research Support, Non-U.S. Gov't
Molecular characterization of mammalian-adapted Korean-type avian H9N2 virus and evaluation of its virulence in mice: Kuk Jin Park ¹, Min-Suk Song ¹, Eun-Ha Kim ¹, Hyeok-il Kwon ¹, Yun Hee Baek ¹, Eun-hye Choi ¹, Su-Jin Park ¹, Se Mi Kim ¹, Young-il Kim ¹, Won-Suk Choi ¹, Dae-Won Yoo ², Chul-Joong Kim ², Young Ki Choi ¹; Journal of Microbiology 2015;53(8):570-577.
DOI: https://doi.org/10.1007/s12275-015-5329-4
Published online: July 31, 2015

¹College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju 361-763, Republic of Korea, ²College of Veterinary Medicine, Chungnam National University, Daejeon 305-764, Republic of Korea¹College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju 361-763, Republic of Korea, ²College of Veterinary Medicine, Chungnam National University, Daejeon 305-764, Republic of Korea

Corresponding author: Young Ki Choi , Tel: +82-43-261-3384,

Received: 29 June 2015 • Accepted: 8 July 2015

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Abstract

Avian influenza A virus (AIV) is commonly isolated from domestic poultry and wild migratory birds, and the H9N2 subtype is the most prevalent and the major cause of severe disease in poultry in Korea. In addition to the veterinary concerns regarding the H9N2 subtype, it is also considered to be the next potential human pandemic strain due to its rapid evolution and interspecies transmission. In this study, we utilize serial lung-to-lung passage of a low pathogenic avian influenza virus (LPAI) H9N2 (A/Ck/Korea/163/04, WT163) (Y439-lineage) in mice to increase pathogenicity and investigate the potential virulence marker. Mouse-adapted H9N2 virus obtained high virulence (100% mortality) in mice after 98 serial passages. Sequence results show that the mouse adaptation (ma163) possesses several mutations within seven gene segments (PB2, PA, HA, NP, NA, M, and NS) relative to the wild-type strain. The HA gene showed the most mutations (at least 11) with one resulting in the loss of an N-glycosylation site (at amino acid 166). Moreover, reverse genetic studies established that an E627K substitution in PB2 and the loss of the N-glycosylation site in the HA protein (aa166) are critical virulence markers in the mouse-adapted H9N2 virus. Thus, these results add to the increasing body of mutational analysis data defining the function of the viral polymerase and HA genes and their roles in mammalian host adaptation. To our knowledge, this is first report of the generation of a mammalian-adapted Korea H9N2 virus (Y493-lineages). Therefore, this study offers valuable insights into the molecular evolution of the LPAI Korean H9N2 in a new host and adds to the current knowledge of the molecular markers associated with increased virulence.

Keywords: influenza A virus;H9N2;mammalian adaptation;reverse genetic method;virulence marker

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Molecular characterization of mammalian-adapted Korean-type avian H9N2 virus and evaluation of its virulence in mice

J. Microbiol. 2015;53(8):570-577. Published online July 31, 2015

DOI: https://doi.org/10.1007/s12275-015-5329-4

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