Development of a new oral rabies vaccine by reverse genetics

本研究以日本狂犬病疫苗株RC-HL为研究对象，对日本狂犬病疫苗株RC-HL进行基因重组，并通过反向遗传学方法获得重组RC-HL株，以生产出对动物具有较高免疫力和安全性的新一代狂犬病疫苗。结果表明：（1）狂犬病毒G蛋白的242、255和268位氨基酸对小鼠的毒力起重要作用;（微生物免疫学，2001; J.Virol.，2001 ; J. Neurovirol.，2）建立了稳定表达T7 RNA聚合酶的BHK细胞克隆，以提高从克隆cDNA中回收狂犬病毒的效率。（微生物免疫学，2003)3)M基因缺陷型狂犬病病毒是从缺失完整M基因的RC-HL株的cDNA产生的。缺陷病毒比灭活疫苗更有效地诱导小鼠产生中和抗体。（2003年在日本病毒学学会发表）4）基因组重排的狂犬病病毒被产生以增强保护性免疫应答。（2004年在日本兽医学会发表）

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)