Development of gene diagnosis and recombinant vaccine for animal morbilliviurs diseases

动物麻疹病毒病基因诊断及重组疫苗的研制

• 批准号: 62480085
• 负责人: YOSHIKAWA Yasuhiro
• 金额: $ 3.65万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62480085/
• 关键词: rinderpest virus; glycoprotein genes; recombinant DNA vaccine; canine distemper; epileptic seizure; viral encephalitis; Japanese monkey; squirrel monkey; 組換えワクチニアウイルスワクチン; バキュロウイルスベクタ-; H遺伝子; NP遺伝子; 合成ペプチド; モノクロ-ナル抗体; エピト-プ解析; H蛋白遺伝子; F蛋白遺

The cDNA library of rinderpest virus (RV) mRNA has been prepared and nucleotide sequence of both glycoprotein genes (H, F protein) has been completely determined.1) Deduced H protein from its nucleotide sequence has a unique structure in which C-terminus projects out of envelope. This predicted structure was confirmed by epitope analysis using ten different types of anti-syntheitc peptide antibodies.2) Deduced F protein has a site of proteolytic cleavage and glycosylation sites on the cleft F2 fragment. This was confirmed by isotope labeling of RV-infected cells.3) Recombinant DNA vaccine consisted of vaccinia virus having RV-H gene showed very strong efficacy in a rabbit model system and complete protection of immunized animals with the recombinant vaccine has been confirmed by clinical, virological and pathological investigations.Neurovirulence of canine distemper virus (CDV) was studied.4) Neural cell-adapted CDV showed relatively high neurovirulence and changed its genome and glycoproteins as a result of adaptation.5) Natural infection and viral encephalitis by CDV in a group of Japanese monkeys were studied. This is the first report in the world that primate has a natural susceptibility to CDV- encephalitis.6) Squirrel monkey which belongs to new world monkeys, was confirmed to have high susceptibility to CDV-encephalitis and to show epilectic seizure during acute encephalitis. It will become an excellent model for viral epilepsy.

建立了牛瘟病毒（RV） mRNA cDNA文库，完整测定了两个糖蛋白基因（H、F蛋白）的核苷酸序列。1)从其核苷酸序列推断出的H蛋白具有独特的结构，其c端在包膜外突出。这一预测结构通过十种不同类型的抗合成肽抗体的表位分析得到证实。2)推导出的F蛋白在裂解的F2片段上有蛋白水解裂解位点和糖基化位点。这是通过rv感染细胞的同位素标记证实的。3)含有RV-H基因的牛痘病毒重组DNA疫苗在家兔模型系统中表现出很强的免疫效果，经临床、病毒学和病理学研究证实，重组疫苗对免疫动物具有完全的保护作用。研究了犬瘟热病毒（CDV）的神经毒力。4)适应神经细胞的CDV表现出较高的神经毒力，并改变了其基因组和糖蛋白。5)对日本猕猴自然感染CDV和病毒性脑炎进行了研究。这是世界上首次报道灵长类动物对CDV-脑炎具有天然易感性。6)松鼠猴属新世界猴，对cvd -脑炎具有高易感性，急性脑炎时表现为癫痫发作。它将成为病毒性癫痫的极好模型。

项目成果

Tsukiyama,K.: "Molecular cloning and sequence analysis of the rinderpest virus mRNA encoding the hemagglutinin protein." Virology. 160. 48-54 (1987)

Tsukiyama,K.：“编码血凝素蛋白的牛瘟病毒 mRNA 的分子克隆和序列分析。”

森川裕子: Microbiol,Immunol. 32. 1263-1268 (1988)

森川裕子：微生物学，免疫学 32. 1263-1268 (1988)

今岡浩一: Arch.Virol. 102. 111-118 (1988)

今冈浩一：Arch.Virol。102. 111-118 (1988)

Tsukiyama,K.: "Fusion glycoprotein(F)of rinderpest virus:Entire nucleotide sequence of the F mRNA, and several feature of the F protein" Vorology. 164. 523-530 (1988)

Tsukiyama,K.：“牛瘟病毒的融合糖蛋白（F）：F mRNA 的完整核苷酸序列，以及 F 蛋白的几个特征”Vorology。

Imaoka, K., Yoshikawa, Y., Kanai, Y., Yamanouchi, K.: "Characterization of antinuclear antibodies induced in rabbits by rinderpest virus infection." Arch. Virol. 102: 111-118 (1988).

Imaoka, K.、Yoshikawa, Y.、Kanai, Y.、Yamanouchi, K.：“牛瘟病毒感染在兔子中诱导的抗核抗体的表征。”