Development of a new oral rabies vaccine by reverse genetics

通过反向遗传学开发新型口服狂犬病疫苗

• 批准号: 13556054
• 负责人: MINAMOTO Nobuyuki
• 金额: $ 4.03万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13556054/
• 关键词: Rabies virus; Revers genetics; New vaccine; Genetic reinforcement; Genetic rearrangement; Immunogenicity; Gene-deficient virus; リバース・ジェネッテクス; 遺伝子配置換え; 免疫原

In this, study, to produce the new generational rabies vaccine enhanced highly immunity and safety for animal, we attempted rearrangement of the genes of the RC-HL strain of rabies virus used for the production of animal vaccine in Japan and rescued several recombinant RC-HL strains by reverse genetics. In addition, we indicated the fundamental informations for creation of recombinant viruses.The results run as follows :1)The three amino acids(242,255 and 268) in the G protein of rabies virus were identified to play an important role in the virulence for mice. (Microbiol.Immunol., 2001; J.Virol., 2001 ; J.Neurovirol., 2004)2)A BHK cell clone expressed stably T7 RNA polymerase was established to improve efficiency of recovery of rabies virus from cloned cDNA. (Microbiol.Immunol., 2003)3)Rabies virus deficient in the M gene was generated from cDNA of RC-HL strain lacking the entire M gene. The defective virus induced neutralizing antibody in mice more effectively than inactivated vaccine. (Presentation in the Japanese Society for Virology, 2003)4)Rabies viruses with rearranged genomes were generated to be enhance the protective immune response. (Presentation in the Japanese Society of Veterinary Science, 2004)

本研究对日本用于动物疫苗生产的狂犬病毒RC-HL株的基因进行了重排，并通过反向遗传学的方法挽救了数株重组RC-HL株，以期生产出对动物具有高免疫力和安全性的新一代狂犬疫苗。此外，我们指出了创建重组病毒的基本信息。结果表明：1)鉴定出狂犬病毒G蛋白中的3个氨基酸（242,255和268）在小鼠毒力中起重要作用。(Microbiol.Immunol。, 2001;J.Virol。， 2001；J.Neurovirol。2)建立了一个稳定表达T7 RNA聚合酶的BHK细胞克隆，提高了从克隆cDNA中回收狂犬病毒的效率。(Microbiol.Immunol。3)由缺失整个M基因的RC-HL株cDNA合成M基因缺失狂犬病毒。缺陷病毒比灭活疫苗更有效地诱导小鼠产生中和抗体。（在日本病毒学学会的报告，2003）4)产生重组基因组的狂犬病毒是为了增强保护性免疫反应。（2004年在日本兽医学会发表的报告）

项目成果

Susetya, H., et al.: "Genetic characterization of rabies field isolates from Thailand."Microbiology & Immunology. 47(9). 653-659 (2003)

Susetya, H. 等人：“泰国狂犬病现场分离株的遗传特征。”微生物学

Ito, N.: "A comparison of complete genome sequences of the attenuated RC-HL strain of rabies vi production of animal vaccine in Japan, and the parental Nishigahara strain"Microbiology &. Immunology. 45(1). 51-58 (2001)

Ito, N.：“日本动物疫苗生产中狂犬病 RC-HL 减毒株与亲本西原株的完整基因组序列的比较”微生物学

Ito, N.: "Rescue of rabies virus from cloned cDNA and identification of the pathogenicity glycoprotein gene is associated with the virulence for adult mice"Journal of Virology. 75(19). 9121-9128 (2001)

Ito, N.：“从克隆的 cDNA 中拯救狂犬病病毒并鉴定致病性糖蛋白基因与成年小鼠的毒力有关”病毒学杂志。

Ito, N.: "Rescue of rabies virus from cloned cDNA and identification of the pathogenicity- related gene : glycoprotein gene is associated with the virulence for adult mice."Journal of Virology. 75(19). 9121-9128 (2001)

Ito, N.：“从克隆的 cDNA 中拯救狂犬病病毒并鉴定致病性相关基因：糖蛋白基因与成年小鼠的毒力有关。”病毒学杂志。

Sugiyama, M.: "Isothermal amplification of rabies virus gene."The Journal Veterinary Medical Science. 65(10). 1063-1068 (2003)

Sugiyama, M.：“狂犬病病毒基因的等温扩增。”《兽医医学科学》杂志。