title: Molecular characterization of mammalian-adapted Korean-type avian H9N2 virus and evaluation of its virulence in mice.
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.
authors.affiliation: College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, 361-763, Republic of Korea.
authors.forename: Kuk Jin
authors.initials: KJ
authors.lastname: Park
authors.forename: Min-Suk
authors.initials: MS
authors.lastname: Song
authors.forename: Eun-Ha
authors.initials: EH
authors.lastname: Kim
authors.forename: Hyeok-Il
authors.initials: HI
authors.lastname: Kwon
authors.forename: Yun Hee
authors.initials: YH
authors.lastname: Baek
authors.forename: Eun-Hye
authors.initials: EH
authors.lastname: Choi
authors.forename: Su-Jin
authors.initials: SJ
authors.lastname: Park
authors.forename: Se Mi
authors.initials: SM
authors.lastname: Kim
authors.forename: Young-Il
authors.initials: YI
authors.lastname: Kim
authors.forename: Won-Suk
authors.initials: WS
authors.lastname: Choi
authors.forename: Dae-Won
authors.initials: DW
authors.lastname: Yoo
authors.forename: Chul-Joong
authors.initials: CJ
authors.lastname: Kim
authors.forename: Young Ki
authors.initials: YK
authors.lastname: Choi
mesh_terms: D000818:Animals; D015415:Biomarkers; D019143:Evolution, Molecular; D005814:Genes, Viral; D053127:Influenza A Virus, H9N2 Subtype / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D008168:Lung / Q000821:virology; D051379:Mice; D009154:Mutation; D009976:Orthomyxoviridae Infections / Q000821:virology*; D056910:Republic of Korea; D014774:Virulence; D014779:Virus Replication / Q000235:genetics
chemical_list: D015415:Biomarkers