| Project/Area Number |
13556046
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Applied veterinary science
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| Research Institution | Hokkaido University |
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Principal Investigator |
MORIMATSU Kumiko (吉松 組子) Hokkaido Univ., Grad. School of Med., Inst., 大学院・医学研究科, 助手 (90220722)
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| Co-Investigator(Kenkyū-buntansha) |
ONO Eriko Merial Japan Limited, Res., つくば明野ラボラトリーズ, 研究員
MORIMATSU Masami Iwate Univ., fac. of Agr., Asso. Prof., 農学部, 助教授 (70241370)
KARIWA Hiroaki Hokkaido Univ., Grad. School of Vet. med., Asso. Prof., 大学院・獣医学研究科, 助教授 (70224714)
ARIKAWA Jiro Hokkaido Univ., Grad. School of Med., Prof., 大学院・医学研究科, 教授 (10142704)
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| Project Period (FY) |
2001 – 2003
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| Keywords | hantavirus / envelope protein / antigenicity / vaccine / diagnosis / purification |
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| Research Abstract |
1. It was clarified that recombinant Hantavirus envelope glycoproteins G1 and G2 (GPs) had the cell-fusion activity by using CAG promotor-expression system. Fusion-responsible epitope on GPs was found to locate in the neutralization (FRNT)-relating epitope. Therefore, the recombinant GPs was considered to possess neutralization-and fusion-relating epitopes. In addition, hantavirus nucleocapsid (N) protein was also expressed in mammalian cells by using same vector. As the result the N protein may suppress that expression and transportation of the envelope protein. It was also shown that the virus like particle was not formed by GPs and N protein alone.
2. By using recombinant GPs, pseudotype vesicular stomatitis virus (VSV) enveloped with hantavirus GPs altered to VSV G protein was produced (VSVΔG*HTN). Similar pseudotype VSV was produced with recombinant GPs of Seoul virus (VSVΔG*SEO). These pseudotypes were applied to rapid neutralization assay as safety alternatives of authentic viruses. These pseudotype virus particles were also applied to vaccination as alternatives to authentic virion. Mice were immunized with soluble recombinant GPs or pseudotype virion. In mice immunized with soluble recombinant GPs, FRNT antibody was not detected. Contrary, in mice immunized with VSVΔG*HTN virion, FRNT antibody was detected. Challenge administration of hantavirus was carried out by s.c. inoculation of 4 FFU of hantavirus. These mice did not show the elevation of anti-N antibody and hantavirus specific CD8 T cell response, indicating that the FRNT antibody could protect mice from hantavirus infection. On the other hand, all control mice immunized with VSVΔG*G or PBS could not escape from hantavirus infection. These results indicated that packaged recombinant GPs on VSV particle were possible to induce protective FRNT antibody. This study shows that novel approach for the development of vaccination of viruses that have difficulties in preparation of authentic virus particles.
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