| Project/Area Number |
61570232
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Virology
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| Research Institution | National Institute of Health |
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Principal Investigator |
MATSUNO Tetsuya Head Investigator,Department of Measles virus, NIH, 麻疹ウィルス部, 厚生技官・室長 (30109970)
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| Co-Investigator(Kenkyū-buntansha) |
UMINO Yukiko Investigator, Department of Measles virus, NIH, 麻疹ウィルス部・厚生技官, 主任研究官 (00223595)
KATOW Shigetaka Investigator, Department of Measles virus, NIH, 麻疹ウィルス部・厚生技官, 主任研究官 (20211162)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1986: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Rubella virus / Monoclonal antibodies / Fusion / Conformational change / Penetration / Uncoating / Lysosome / 中和(NT)リポソーム / 赤血球凝集(HA) / 中和 / 赤血球溶血 |
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| Research Abstract |
1, We have immunized mice with rubella virus and obtained fifteen monoclonal antibodies to E1 glycoprotein and two monoclonal antibodies to C protein. Four antibodies were found to react with a new antigenic site which differs from the three sites on E1, reported previously (Umino et al, 1985). Neither the anti-El antibodies, nor the anti-C antibodies showed biological activities.
2, Fusion of rubella virus-infected cells was induced by their brief treatment at pH below 6.0. Exposure of rubella virus of pH 5 caused an irreversible conformational change of the viral envelope glycoproteins, E1 and E2. The change was manifested in the marked reduction in both infectivity and hemagglutinating activity of the virus, the increased resistance of E1 and decreased resistance of E2 polypeptides to proteolytic digestion with trypsin, and the acquisition of liposome-binding activity of the vius. The above changes are presumed to mimic the events occurring in the acidic environment within endosomes following endocytosis of virus.
3, We examined the antigenic reactivities and the relationship with virions of glycoproteins that were released into the culture fluids of BNK21 cells infected with rubella virus. The culture fluids of radiolabeled, virus-infected cells were fractionated by ultracentrifugation into pellet and supernatant. The former was found to be viroins and the latter was concentrated using ammonium sulfate. This fraction was further subjected to rate zonal centrifugation through a 5 to 20% sucrose gradient. An assay of fractions by immunoprecipitation and SDS-PAGE revealed the presence of a 42K glycoprotein which has been identified as E1. This glycoprotein was smaller than E1 on the virion (MW 58K). These results suggested that the E1 glycoprotein is released into the culture fluid from infected cells by proteolysis.
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