Plasmid DNA launches live-attenuated Japanese encephalitis virus and elicits virus-neutralizing antibodies in BALB/c mice.
Plasmid DNA launches live-attenuated Japanese encephalitis virus and elicits virus-neutralizing antibodies in BALB/c mice.
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DOI：
10.1016/j.virol.2017.09.005
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发表时间：
2017-12
期刊：
影响因子：
3.7
通讯作者：
Pushko P
中科院分区：
文献类型：
--
作者：
Nickols B;Tretyakova I;Tibbens A;Klyushnenkova E;Pushko P
我们描述了编码乙型脑炎病毒(JEV)全长基因组DNA的新型质粒DNA，并在体内外推出了JEV减毒活疫苗。根据乙型脑炎病毒SA14-14-2减毒活病毒序列合成的cDNA置于巨细胞病毒主要即刻早期启动子的转录控制下。通过在结构基因和非结构基因中插入三个扰乱JEV基因的合成内含子，优化了该质粒在大肠杆菌中的稳定性和产量。用所得到的质粒转染Vero细胞，可以复制病毒RNA中去除了内含子序列的乙脑疫苗病毒。此外，用0.5~5μg的重组质粒单次免疫BALB/c小鼠，可成功地产生乙型脑炎病毒中和血清抗体。研究结果表明，利用DNA疫苗研制乙脑减毒活疫苗是可行的，并为进一步研制预防乙脑感染的DNA减毒活疫苗奠定了基础。
We describe novel plasmid DNA that encodes the full-length Japanese encephalitis virus (JEV) genomic cDNA and launches live-attenuated JEV vaccine in vitro and in vivo. The synthetic cDNA based on the sequence of JEV SA14-14-2 live-attenuated virus was placed under transcriptional control of the cytomegalovirus major immediate-early promoter. The stability and yields of the plasmid in E. coli were optimized by inserting three synthetic introns that disrupted JEV cDNA in the structural and nonstructural genes. Transfection of Vero cells with the resulting plasmid resulted in the replication of JEV vaccine virus with intron sequences removed from viral RNA. Furthermore, a single-dose vaccination of BALB/c mice with 0.5 – 5 μg of plasmid resulted in successful seroconversion and elicitation of JEV virus-neutralizing serum antibodies. The results demonstrate the possibility of using DNA vaccination to launch live-attenuated JEV vaccine and support further development of DNA-launched live-attenuated vaccine for prevention of JEV infections.
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影响因子：
3.7
作者：
Saito Y;Moi ML;Takeshita N;Lim CK;Shiba H;Hosono K;Saijo M;Kurane I;Takasaki T
通讯作者：
Takasaki T
影响因子：
6.2
作者：
Pushko P;Lukashevich IS;Weaver SC;Tretyakova I
通讯作者：
Tretyakova I
影响因子：
5.4
作者：
Pushko, P;Geisbert, J;Smith, J
通讯作者：
Smith, J
影响因子：
5.5
作者：
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通讯作者：
Bernard, R
影响因子：
2.6
作者：
Bharati, K;Appaiahgari, MB;Vrati, S
通讯作者：
Vrati, S