Challenges to replicate human norovirus with tissue cells

• Project/Area Number: 23656325
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Civil and environmental engineering
• Research Institution: Tohoku University
• Principal Investigator: OMURA Tatsuo 東北大学, 大学院・工学研究科, 教授 (30111248)
• Co-Investigator(Kenkyū-buntansha): SANO Daisuke 北海道大学, 大学院・工学研究院, 准教授 (80550368) ; MASAGO Yoshifumi 東北大学, 大学院・工学研究科, 助教 (50507895)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: ヒトノロウイルス / 培養 / 組織細胞 / ノロウイルス / ウイルス様中空粒子 / 緑色蛍光タンパク質 / ウイルス / 感染症

Research Abstract

The main purpose of this project was to acquire precious knowledge for establishing the infection of human norovirus to tissue cells. We successfully constructed a norovirus-like particle with green fluorescence protein (GFP-NoVLP). GFP-NoVLP was applied for confirming the intrusion and decapsidation of human norovirus in tissue cells, but it was found that the amount of fluorescence was not enough to observe the movement of GFP-NoVLP in tissue cells. Therefore, we paid attentions to cellular responses of tissue cells against the virus infection, and tried to identify genetic markers available for the detection of virus infection. Poliovirus type 1 (PV1) Sabin strain was used as a test virus, and a transcriptome analysis of INT407 cells infected with PV1 was conducted. As a result, KCNJ4 gene, responsible for potassium homeostasis, was found as the most promising genetic marker for the detection of virus infection.

Research Products

• [Journal Article] A new approach for evaluating the infectivity of noncultivatable enteric viruses without cell culture (2013)
  • Author(s): Tojo, K., Sano, D., Miura, T., Nakagomi, T., Nakagomi, O. and Okabe, S.
  • Journal Title: Water Science and Technology Volume: 67 Issue: 10 Pages: 2236-2240
  • DOI: 10.2166/wst.2013.114
  • NAID: 120005358932
  • Peer Reviewed
• [Journal Article] A new approach for evaluating the infectivity of nonculturable enteric viruses without cell culture (2013)
  • Author(s): Kazuki Tojo
  • Journal Title: Water Science and Technology Volume: In press
  • Peer Reviewed
• [Presentation] Microbiologically safe water management in membrane-based wastewater reuse (2012)
  • Author(s): Daisuke Sano and Satoshi Okabe
  • Organizer: 1st Symposium on Innovation and Water Monitoring System with Rapid, Highly Precise and Exhaustive Pathogen Detection Technologies
  • Place of Presentation: Sendai, Japan
  • Year and Date: 2012-10-20
• [Presentation] Transcriptome analysis of intestinal epithelial cells infected with poliovirus type 1 (2012)
  • Author(s): Megumi Tazawa, Takayuki Miura, Daisuke Sano and Satoshi Okabe
  • Organizer: Europic 2012
  • Place of Presentation: Saint Paphael, France
• [Presentation] 緑色蛍光タンパク質付加ノロウイルス様中空粒子 (2012)
  • Author(s): 山口諒、今井崇博、西牧宏尚、真砂佳史、大村達夫
  • Organizer: 第46回水環境学会年会
  • Place of Presentation: 東京
• [Presentation] Transcriptome analysis of intestinal epithelial cells infected with poliovirus type 1 (2012)
  • Author(s): Megumi Tazawa
  • Organizer: Europic 2012
  • Place of Presentation: Saint Raphael, France.
• [Presentation] 緑色蛍光タンパク質付加ノロウイルス様中空粒子 (2012)
  • Author(s): 山口諒，他
  • Organizer: 第44回日本水環境学会年会
  • Place of Presentation: 東京
• [Presentation] Microbiologically safe water management in membrane-based wastewater reuse
  • Author(s): Daisuke Sano
  • Organizer: 1st symposium on innovation of water monitoring system with rapid, highly precise and exhaustive pathogen detection technologies
  • Place of Presentation: Sendai, Japan