Li Wen-hui, Zhang Yun, Wang Shu-hui, Liu Li, Yang Fan
Department of Etiology, Institute of Basic Medical Science, CAMS and PUMC, Beijing 100005, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2003 Dec;25(6):650-4.
To construct and characterize recombinant adenoviruses containing glycoprotein (GP) gene from rabies virus CVS-N2C strain.
To obtain the recombinant adenovirus by pAdEasy system, identify recombinant virus with cDNA sequencing, Northern blot, Dot blot, Western blot and challenge-protection experiment of mice.
Recombinant adenovirus showed typical adenovirus morphological characteristics; the viral genome was stable; GP specific mRNA and presence of glycoprotein were determined in rAdGPcvs and rAdGPcvs' infected cells. The glycoprotein produced by recombinant adenovirus had a molecular mass of 66,000, which was similar to that of natural glycoprotein. In the group of rAdGPcvs immunized mice, 87.5%-100% of mice survived from a 35.8LD50/38.0LD50 lethal rabies intracerebral challenge. Finally 73.3%-83.3% of the mice that had received eAdGPcvs survived, and all the Ad5 immunized mice succumbed to rabies.
Recombinant adenovirus rAdGPcvs and rAdGPcvs' hold great potential to be developed as recombinant rabies vaccines, and at the same time, it is actually the first study that on high neuropathogenicity rabies CVS-N2C glycoprotein based adenoviral recombinants.
构建并鉴定含狂犬病病毒CVS-N2C株糖蛋白（GP）基因的重组腺病毒。
采用pAdEasy系统获得重组腺病毒，通过cDNA测序、Northern印迹、斑点印迹、Western印迹及小鼠攻毒保护实验对重组病毒进行鉴定。
重组腺病毒呈现典型的腺病毒形态特征；病毒基因组稳定；在rAdGPcvs和rAdGPcvs'感染的细胞中检测到GP特异性mRNA及糖蛋白的存在。重组腺病毒产生的糖蛋白分子量为66000，与天然糖蛋白相似。在rAdGPcvs免疫的小鼠组中，87.5%-100%的小鼠在接受35.8LD50/38.0LD50致死量的狂犬病脑内攻毒后存活。最后，接受eAdGPcvs的小鼠中有73.3%-83.3%存活，而所有接种Ad5的小鼠均死于狂犬病。
重组腺病毒rAdGPcvs和rAdGPcvs'具有开发成为重组狂犬病疫苗的巨大潜力，同时，这实际上是首次对基于高神经致病性狂犬病CVS-N2C糖蛋白的腺病毒重组体进行的研究。
