Grant-in-Aid for Developmental Scientific Research.
|Research Institution||Tokyo metropolitan Institute for Neurosciences|
YASUI Kotaro (財)東京都神経科学総合研究所, 微生物学研究部門, 副参事研究員 (90073080)
小島 朝人 国立予防衛生研究所, 病理部, 室長 (30100077)
松浦 善治 国立予防衛生研究所, 獣疫部, 厚生技官 (50157252)
宮本 道子 (財)東京都神経科学総合研究所, 微生物学研究部門, 主事研究員 (40190821)
荻本 真美 (財)東京都神経科学総合研究所, 微生物学研究部門, 主事研究員 (80158609)
木村 純子 (財)東京都神経科学総合研究所, 微生物学研究部門, 主任研究員 (20142151)
竹上 勉 金沢医科大学, 総合医学研究所, 助教授 (10113490)
|Project Fiscal Year
1989 – 1991
Completed(Fiscal Year 1991)
|Budget Amount *help
¥19,000,000 (Direct Cost : ¥19,000,000)
Fiscal Year 1991 : ¥2,700,000 (Direct Cost : ¥2,700,000)
Fiscal Year 1990 : ¥3,900,000 (Direct Cost : ¥3,900,000)
Fiscal Year 1989 : ¥12,400,000 (Direct Cost : ¥12,400,000)
|Keywords||Japanese encephalitis virus / E Protein / new vaccine / recombinant virus / Flaviviruses / JEV / HBsAg with JEV E / 日本脳炎ウイルス / フラビウイルス / E蛋白 / 新ワクチン / 組換えウイルス / HBsAg粒子 / 粒子 / 日本脳炎ウィルス / 組換えウィルス / フラビウィルス / HBsAg / 新型ワクチン / ワクチン / 抗原構造 / 組換えバキュロウィルス / 組換えワクチニアウィルス|
Recombinant baculo- and vaccinia viruses containing the coding sequences of the viral envelope proteins, i. e., preill, M, and E protein of Japanese encephalitis virus (JEV) were constructed. These recombinants which had a signal sequence at the upstream of each coding sequence of the proteins produced biologically and antigenica-lly active JEV proteins which were glycosylated and processed as similar to the authentic proteins. Our results also indicate that efficient expression of the E protein by recombinant virus requires the signal sequence which is encoded upstream of E protein and when each epitope of E protein is successfully expressed, this protein is detected on surfaces and culture supernatant of recombinant virus infected cells. The E proteins released from recombinant virus infected cells were small particles and oligomers or heterooligomers with M protein.
Animals infected with these recombinants or received the product of them showed similar immune response against these p
roteins including remarkable production of neutralizing antibodies as JEV infection and challenge testing was successfully done by using mice. The E protein oligomer and hetero oligomer particles composed with M protein were more good immunogens and elicited biologically active antibodies such as neutraling and hemaggulutination inhibition than monomer E.
Locations of JEV specific and WN subgroup virus specific neutralizing epitopes and strain specific epitopes on the E protein were detenaned by analyzing monoclonal escaped mutants and strains of JEV. HBsAgs fused with a part of JEV E protein containing only specific epitopes were successfully produced in particulate forms.
As a conclusion, we could develop the methods which could produce particulate E proteins and also HBsAg particles with only specific epitopes to induce neutralizing antibody effectively. From these results we could show the basic informations to develop new vaccines for JEV and other flaviviruses by recombinant virus technologies. Less