1994 Fiscal Year Final Research Report Summary
Development of poliovirus-rotavirus chimeric vaccine by using poliovirus artificial interfering particle RNA
| Project/Area Number |
04557025
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Virology
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| Research Institution | SAPPORO MEDICAL UNIVERSITY |
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Principal Investigator |
TANIGUCHI Koki Sapporo Medical University School of Medicine, Associate Professor, 医学部, 助教授 (40094213)
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| Co-Investigator(Kenkyū-buntansha) |
AIZAWA Chikara The Kitasato Institute, Research Center for Biologicals, Director, 生物製剤研究所, 所長 (80072362)
KUGE Shusuke The University of Tokyo, The Institute of Medical Science, Instructor, 医科学研究所, 教務職員 (50186376)
YAMAGISHI Kimiko The Tokyo Metropolitan Institute of Medical Science, Researcher, 微生物部, 研究員 (20200602)
KOBAYACHI Nobumichi Sapporo Medical University School of Medicine, Assistant Professor, 医学部, 講師 (80186759)
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| Project Period (FY) |
1992 – 1994
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| Keywords | Rotavirus / Poliovirus / Vaccine / VP4 gene / DI particle / Cross-reactive neutralization epitope |
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| Research Abstract |
We constructed polio-rota VP4 chimeric cDNA by inserting the DNA fragments encoding VP8' or VP5' into pSM1 (T7) 3 or pSM1 (T7) 2 vector having poliovirus artificial DI genome. In the HeLa cells transfected with polio-rota chimeric RNA obtained from pSM1 (T7) 3-VP5', pSM1 (T7) 3-VP8', or pSM1 (T7) 2-VP8'+VP5', replication of rotavirus-specific RNA could be detected. However, the expression of rotaviral protein was observed only in the cells transfected with pSM1 (T7) 3-VP8' or pSM1 (T7) 2-VP8'+VP5'. The antigenicity of the expressed VP5' might be weak in this system because of the conformational change in the from of the VP5' fused with polioviral protein. Rotaviral protein reactive with antiserum to VP4 could be detected in the cells at the fourth generation after transfection with pSM1(T7)3-VP8' and infection with infectious poiliovirus. By preparing of polio-rota chimeric virus with high titer and by using the transgenic mice with poliovirus receptor which have the potency of oral inoculation, the immunogenicity of the poliorota chimeric virus would be investigated.
We determined the nucleotide and deduced amino acid sequences of the VP4 genes of the rotaviruses from various mammalian species including humans. Comparative analysis of the VP4 sequences showed that there are at least 17 kinds of VP4 genotypes and that species specificity and interspecies relatedness in VP4 geneotypes are present. It was also shown that some human rotaviruses have VP4 geneotypes related to those of animal rotaviruses. These findings should be taken into considerataion for rotavirus vaccine development.
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