DEVELOPMENT OF NEW MUMPS VIRUS VACCINE BY USING GENE MANIPULATION

利用基因操作开发新型腮腺炎病毒疫苗

• 批准号: 06557021
• 负责人: ITO Yasuhiko
• 金额: $ 4.61万
• 依托单位: MIE UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557021/
• 关键词: MUMPS VIRUS; PARAINFLUENZA VIRUS; REVERSE GENETICS; VACCINE; リバースジェネティクス

For development of new Mumps virus vaccine, we have tried to establishe the reverse genetice of parainfluenza viruses, especially HPIV-2. We examined two systems, that is, helper virus system and Vaccinia virus-T7 system. After a disperate effort we managed to succeed to some extent. Furthermore, we investigated basic virology on paramyxovirus and clarified the following results ; Interaction of the NP and V proteins of HPIV-2 was investigated. When the NP protein was co-expressed with the V protein, a part of NP proteins was translocated into the nuclei. These findings suggested that the NP proteins interact with the V proteins. We examined interaction of the NP protein and the P,V protein or deletion mutants of V protein by immunofluorescent and co-immunoprecipitation + Western blotting analyzes. Co-expressing with the V protein or the N-terminal fragment (a.a.1 to 46) , the NP protein was detected diffusely in the nuclei of the transfected cells, and was also detected in cytoplasmic inclusions, and the NP protein was coprecipitated with the P,V,and V (1-164) proteins by specific antibody. These findings indicate that the V proteins have an ability to bind the NP proteins. At present we are preparing full-length cDND clones for parainfluenza type 2 virus and HPIV-2-MuV chimera. We think that development of new mumps virus vaccine will be prepared in the future by gene engineering technique established by this study

为了研制新型腮腺炎病毒疫苗，我们尝试建立副流感病毒，特别是HPIV-2的反向遗传学。我们研究了两个系统，即辅助病毒系统和牛痘病毒-T7系统。经过分散的努力，我们总算取得了一定的成功。此外，我们研究了副粘病毒的基本病毒学，并澄清了以下结果：研究了HPIV-2的NP和V蛋白的相互作用。当NP蛋白与V蛋白共表达时，部分NP蛋白被转运到细胞核中。这些发现表明NP蛋白与V蛋白相互作用。我们通过免疫荧光和免疫共沉淀+ Western印迹分析来检测NP蛋白与P、V蛋白或V蛋白缺失突变体的相互作用。与V蛋白或N-末端片段（a.a.1 - 46）共表达的NP蛋白在转染细胞的细胞核中弥漫地检测到，并且还在胞质内含物中检测到，并且NP蛋白通过特异性抗体与P、V和V（1-164）蛋白共沉淀。这些发现表明V蛋白具有结合NP蛋白的能力。目前，我们正在制备2型副流感病毒和HPIV-2-MuV嵌合体的全长cDNA克隆。我们认为，本研究建立的基因工程技术将为今后研制新型腮腺炎病毒疫苗奠定基础

项目成果

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Hide Kan：“融合调节蛋白（FRP）-1/4F2相关分子的鉴定”细胞结构和功能20. 473-483（1996）。

M.Tsurudome, M.Kawano, T.Yuasa, N.Tabata, M.Nishio, H.Komada, and Y.Ito: "Characterization of Bangor Virus Proteins by Using Monoclonal Antibodies" Avian Disease. in press.

M.Tsurudome、M.Kawano、T.Yuasa、N.Tabata、M.Nishio、H.Komada 和 Y.Ito：“使用单克隆抗体表征班戈病毒蛋白”禽病。

S.Suga, M.Tsurudome, S.Ohgimoto, N.Tabata, N.Watanabe, M.Nishio, M.Kawano, H.Komada, , and Y.Ito: "Intracellular localization of antigens recognized by anti-vimentin monoclonal antibodies (mAb) : cross-reactivities of anti-vimentin mAbs with other cellula

S.Suga、M.Tsurudome、S.Ohgimoto、N.Tabata、N.Watanabe、M.Nishio、M.Kawano、H.Komada、和 Y.Ito：“抗波形蛋白单克隆抗体识别的抗原的细胞内定位

湯浅徹 他: "A Cell Fusion-inhibiting Monoclonal Antibody Binds to the Presumed Stalk Domain of the Human Parainfluenza Type 2 Virus Hemagglutinin-Neuraminidase Protein." Virology. 206. 1117-1125 (1995)

Toru Yuasa 等人：“细胞融合抑制单克隆抗体与人副流感 2 型病毒血凝素神经氨酸酶蛋白的假定茎结构域结合。病毒学”206。1117-1125。

Machiko Nishio: "Fusion properties of cells constitutively expressing human parainfluenza type 4A HN and F virus glycoproteins。" Journal of General Virology. 75. 3517-3523 (1994)

Machiko Nishio：“组成型表达人副流感 4A 型 HN 和 F 病毒糖蛋白的细胞的融合特性。普通病毒学杂志 75。3517-3523 (1994)