The attenuation of vaccinia Tian Tan strain by the removal of the viral M1L-K2L genes.
The attenuation of vaccinia Tian Tan strain by the removal of the viral M1L-K2L genes.
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DOI：
10.1016/j.jviromet.2007.03.012
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发表时间：
2007-09
影响因子：
3.1
通讯作者：
Chen Z
中科院分区：
文献类型：
--
作者：
Zhu W;Fang Q;Zhuang K;Wang H;Yu W;Zhou J;Liu L;Tien P;Zhang L;Chen Z
为了获得安全的天坛痘苗病毒疫苗载体，需要开发一种减毒的方法。通过用GFP基因替换病毒M1L-K2 L基因来构建修饰的VTT（MVTT 2-GFP）。与亲本VTT相比，MVTT 2-GFP在兔RK 13和人HeLa细胞系中失去了其复制能力。电镜观察表明，在这两种细胞系中，病毒复制的生命周期在不同阶段被阻断。通过测量鼻内病毒接种后的小鼠体重减轻，MVTT 2-GFP的毒性比VTT低100倍，通过测定小鼠颅内LD 50值，MVTT 2-GFP的毒性比VTT低340倍。外源GFP基因在Vero细胞中传代10轮后，遗传稳定。重要的是，MVTT 2-GFP在小鼠中引起对GFP基因的体液和细胞介导的免疫应答。用105 PFU病毒的两次肌内接种，通过ELISA测定，抗GFP抗体倒数终点滴度达到700以上。针对GFP的CD 8 + T细胞特异性表位，分泌IFN-γ的T细胞的数量达到每百万个脾细胞超过350 SFU。总的来说，去除M1L-K2 L基因是产生减毒痘苗天坛疫苗载体的有用方法。
To generate a safe vaccinia Tian Tan (VTT)-based vaccine vector, it is necessary to develop a method to attenuate the virus. A modified VTT (MVTT2-GFP) was constructed by replacing the viral M1L-K2L genes with a GFP gene. In comparison to the parental VTT, MVTT2-GFP lost its replication capacity in rabbit RK13 and human HeLa cell lines. The life cycle of viral replication was blocked at different stages in these two cell lines as determined by electron microscope examination. MVTT2-GFP was less virulent than VTT for 100-fold by measuring mouse body weight loss after intranasal viral inoculation and for 340-fold by determining the intracranial LD50 value in mice. The foreign GFP gene was stable genetically after 10 rounds of passage in Vero cells. Importantly, MVTT2-GFP elicited both humoral and cell-mediated immune responses to the GFP gene in mice. With two intramuscular inoculations of 105 PFU virus, the anti-GFP antibody reciprocal endpoint titer reached over 700 as determined by an ELISA. The number of IFN-γ secreting T cells reached over 350 SFU per million splenocytes against a CD8+ T cell-specific epitope of GFP. Collectively, the removal of the M1L-K2L genes is a useful method to generate an attenuated vaccinia Tian Tan vaccine vector.
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DOI：
10.1073/pnas.93.21.11341
发表时间：
1996-10-15
影响因子：
11.1
作者：
Moss, B
通讯作者：
Moss, B
影响因子：
3.5
作者：
Antoine, G;Scheiflinger, F;Dorner, F
通讯作者：
Dorner, F
影响因子：
3.7
作者：
Carroll, MW;Moss, B
通讯作者：
Moss, B
影响因子：
2.7
作者：
Chakrabarti, S;Sisler, JR;Moss, B
通讯作者：
Moss, B
影响因子：
4.4
作者：
Jing, LC;Chong, TM;Koelle, DM
通讯作者：
Koelle, DM