Production of influenza virus-like particles in recombinant insect cells

2018 Fiscal Year Final Research Report

• Project/Area Number: 15H04195
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Kobe University
• Principal Investigator: Yamaji Hideki 神戸大学, 工学研究科, 教授 (40283874)
• Research Collaborator: KATSUDA Tomohisa
• Project Period (FY): 2015-04-01 – 2019-03-31
• Keywords: 昆虫細胞 / 組換えタンパク質生産 / ウイルス様粒子 / インフルエンザウイルス / ワクチン

Outline of Final Research Achievements

Production of influenza virus-like particles in recombinant insect cells was investigated. The cDNA fragments encoding hemagglutinin (HA) and matrix protein 1 (M1) of an influenza virus A were separately cloned into the plasmid vector pIHAbla and pIHAneo. The pIHAbla and pIHAneo contained the Bombyx mori actin promoter downstream of the B. mori nucleopolyhedrovirus (BmNPV) IE-1 transactivator and the BmNPV HR3 enhancer for high-level expression, together with either a blasticidin or neomycin resistance gene for use as a selectable marker, respectively. After cotransfection with the prepared plasmids, High Five cells were incubated with blasticidin and G418, and cells resistant to the antibiotics were effectively obtained. Sucrose density-gradient sedimentation analysis of the culture supernatant showed that secreted HA and M1 molecules were produced in a particulate form.

Free Research Field

生物化学工学

Academic Significance and Societal Importance of the Research Achievements

複数の構造タンパク質とエンベロープから構成されるインフルエンザウイルス様粒子の組換え昆虫細胞による生産については，これまで報告されていない．本研究で構築した基盤技術は，亜型の異なるインフルエンザウイルスのウイルス様粒子ワクチンを開発・生産する際にも利用することができる．気候変動，グローバル化，高齢化などにより感染症の深刻化が懸念される状況のもと，本研究で得られた成果はインフルエンザなどのウイルス感染症から人類を守る一助になると期待される．