Establishment of novel HCV cell-culture system using HCV strains other than JFH-1

2013 Fiscal Year Final Research Report

• Project/Area Number: 23790513
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Virology
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: MURAYAMA Asako 国立感染症研究所, ウイルス第二部, 協力研究員 (40415534)
• Project Period (FY): 2011 – 2012
• Keywords: ウイルス / 細胞培養 / ゲノム複製

Research Abstract

We established a novel HCV genotype 2a cell-culture system using J6CF strain harboring five mutations in NS4A, NS5B and 3'UTR. We also established an HCV genotype 2b cell-culture system using a chimeric genome of MA strain harboring minimal regions of JFH-1. These cell-culture systems will be useful for characterizing genotype 2a and 2b viruses and developing antiviral strategies. Our data provides information that will prove essential for the establishment of replication-competent variants of HCV strains that are currently replication incompetent in cultured cells.

Research Products

• [Journal Article] Single strain isolation method for cell culture-adapted hepatitis C virus by end-point dilution and infection (2014)
  • Author(s): Sugiyama N, Murayama A, Suzuki R, Watanabe N, Shiina M, Liang TJ, Wakita T, Kato T
  • Journal Title: PLoS One Volume: 9(5) Pages: e98168
• [Journal Article] NS5B-RNAポリメラーゼの構造と機能 (2013)
  • Author(s): 村山麻子、加藤孝宣
  • Journal Title: 肝胆膵 Volume: 67(6) Pages: 918-923
• [Journal Article] Japanese reference panel of blood specimens for evaluation of hepatitis C virus RNA and core antigen quantitative assays (2012)
  • Author(s): Murayama A, Sugiyama N, Watashi K, Masaki T, Suzuki R, Aizaki H, Mizuochi T, Wakita T, Kato T
  • Journal Title: J Clin Microbiol Volume: 50(6) Pages: 1943-1949
• [Journal Article] Novel cell culture-adapted genotype 2a hepatitis C virus infectious clone (2012)
  • Author(s): Date T, Kato T, Kato J, Takahashi H, Morikawa K, Akazawa D, Murayama A, Tanaka-Kaneko K, Sata T, Tanaka Y, Mizokami M, Wakita T
  • Journal Title: J Virol Volume: 86(19) Pages: 10805-10820
• [Journal Article] A Subclone of HuH-7 with Enhanced Intracellular Hepatitis C Virus Production and Evasion of Virus Related-Cell Cycle Arrest (2012)
  • Author(s): Murayama A, Sugiyama N, Yoshimura S, Ishihara-Sugano M, Masaki T, Kim S, Wakita T, Mishiro S, Kato T
  • Journal Title: PLoS One Volume: 7(12) Pages: e52697
• [Journal Article] Production of Infectious Chimeric Hepatitis C Virus Genotype 2b Harboring Minimal Regions of JFH-1 (2012)
  • Author(s): Murayama A, Kato T, Akazawa D, Sugiyama N, Date T, Masaki T, Nakamoto S, Tanaka Y, Mizokami M, Yokosuka O, Nomoto A, Wakita T
  • Journal Title: J Virol Volume: 86(4) Pages: 2143-2152
• [Presentation] An adaptive mutation in NS4A region enhances Hepatitis C Virus replication and virus production in cell culture (2013)
  • Author(s): 村山麻子、杉山奈央、脇田隆字、加藤孝宣
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 20131203-06
• [Presentation] HCV NS5Aキメラウイルスを用いた第二世代NS5A阻害剤の抗ウイルス活性の評価 (2013)
  • Author(s): 村山麻子、杉山奈央、脇田隆字、加藤孝宣
  • Organizer: 第61回日本ウイルス学会学術集会
  • Place of Presentation: 神戸
  • Year and Date: 20131110-12
• [Presentation] Evaluation of second-generation NS5A inhibitors against hepatitis C virus by using NS5A replaced JFH-1 viruses and full-length infectious clones other than JFH-1 (2013)
  • Author(s): Murayama A, Sugiyama N, Wakita T, Kato T
  • Organizer: 64th Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: Washington D.C., USA
  • Year and Date: 20131102-05
• [Presentation] Assessment of antiviral activities of NS5A inhibitors on recombinant HCV replaced with NS5A from genotypes 1 and 2 strains (2013)
  • Author(s): Murayama A, Sugiyama N, Masaki T, Wakita T, Kato T
  • Organizer: 20th International Symposium on Hepatitis C and Related Viruses
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 20131006-10
• [Presentation] Identification of cell-culture adapted hepatitis C virus JFH-1 variants by single virus isolation with end-point dilution method (2013)
  • Author(s): Sugiyama N, Murayama A, Suzuki R, Shiina M, Wakita T, Kato T
  • Organizer: 20th International Symposium on Hepatitis C and Related Viruses
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 20131006-10
• [Presentation] NS5A領域置換HCVキメラ株を用いたNS5A阻害剤の株特異的抗ウイルス活性の評価 (2012)
  • Author(s): 村山麻子、杉山奈央、岡本有加、政木隆博、脇田隆字、加藤孝宣
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 20121211-14
• [Presentation] C型肝炎ウイルス遺伝子型2b株とJFH-1株のキメラウイルスを用いた抗ウイルス薬評価系の樹立 (2012)
  • Author(s): 村山麻子、加藤孝宣、杉山奈央、脇田隆字
  • Organizer: 第60回日本ウイルス学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 20121113-15
• [Presentation] Efficient hepatitis C virus production associated with enhanced virus assembly and evasion of cell cycle arrest (2012)
  • Author(s): Murayama A, Sugiyama N, Yoshimura S, Ishihara-Sugano M, Wakita T, Mishiro S, Kato T
  • Organizer: 63rd Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: Boston, USA
  • Year and Date: 20121109-13
• [Presentation] Infectious Virus Production with Hepatitis C Virus Genotype 2b Genome Harboring Minimal Regions of JFH-1 (2012)
  • Author(s): Murayama A, Kato T, Sugiyama N, Wakita T
  • Organizer: 19th International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Venice, Italy
  • Year and Date: 20121005-09
• [Presentation] C型肝炎ウイルス産生効率の良いHuH-7細胞サブクローンの同定と解析 (2012)
  • Author(s): 村山麻子、三代俊治、脇田隆字、加藤孝宣
  • Organizer: 第19回肝細胞研究会
  • Place of Presentation: 札幌
  • Year and Date: 20120629-30
• [Presentation] HuH-7 subclone that supports high HCV production due to high virus assembly (2011)
  • Author(s): Murayama A, Sugiyama N, Yoshimura S, Ishihara- Sugano M, Wakita T, Kato T
  • Organizer: International Union of Microbiological Societies 2011 Congress. XV International Congress of Virology
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 20110911-16
• [Presentation] Strain Specific Susceptibility to The Hepatitis C Virus NS5A Inhibitor (2011)
  • Author(s): Okamoto Y, Masaki T, Murayama A, Nomoto A, Wakita T, Kato T
  • Organizer: International Union of Microbiological Societies 2011 Congress. XV International Congress of Virology
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 20110911-16
• [Presentation] Efficient HCV production system using HuH-7 subclone with high virus assembly efficiency (2011)
  • Author(s): Murayama A, Sugiyama N, Yoshimura S, Ishihara- Sugano M, Wakita T, Kato T
  • Organizer: 18th International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, USA
  • Year and Date: 20110908-12
• [Presentation] Development of recombinant hepatitis C virus with NS5A from strains of genotypes 1 and 2 : virus production and susceptibility to NS5A inhibitor (2011)
  • Author(s): Okamoto Y, Masaki T, Murayama A, Wakita T, Kato T
  • Organizer: 18th International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, USA
  • Year and Date: 20110908-12
• [Patent(Industrial Property Rights)] C型肝炎ウイルスJ6CF株ゲノム由来の変異体レプリコン(MUTANT REPLICON DERIVED FROM GENOME OF HEPATITIS C VIRUS J6CF STRAIN) (2012)
  • Inventor(s): 脇田隆字、村山麻子、加藤孝宣
  • Industrial Property Rights Holder: 脇田隆字、村山麻子、加藤孝宣
  • Industrial Property Rights Type: PCT/P2012/064070
  • Industrial Property Number: WO2012/165542
  • Filing Date: 2012-05-31
  • Overseas