Nakatsu Yuichiro, Takeda Makoto, Kidokoro Minoru, Kohara Michinori, Yanagi Yusuke
Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan.
J Virol Methods. 2006 Oct;137(1):152-5. doi: 10.1016/j.jviromet.2006.05.029. Epub 2006 Jul 18.
A rescue system for measles virus from cloned cDNA was established using CHO/hSLAM cells (Chinese hamster ovary cells expressing a measles virus receptor, signaling lymphocyte activation molecule), LO-T7-1 virus (the Lister vaccine strain of vaccinia virus expressing the T7 RNA polymerase under the control of the early/late p7.5 promoter), and caspase inhibitor. LO-T7-1 drove efficiently the T7 promoter in CHO/hSLAM cells. Rescue efficiency with LO-T7-1 was not as high as that with the vTF7-3 strain based on a neurovirulent vaccinia virus, but was increased by using a caspase inhibitor to block apoptosis of CHO/hSLAM cells induced by LO-T7-1. These modifications resulted in a measles virus rescue efficiency that was even higher than that of previous systems. This safer and more efficient system will facilitate further the genetic manipulation of measles virus in basic research and virus vector development.
利用CHO/hSLAM细胞(表达麻疹病毒受体——信号淋巴细胞激活分子的中国仓鼠卵巢细胞)、LO-T7-1病毒(在早期/晚期p7.5启动子控制下表达T7 RNA聚合酶的痘苗病毒李斯特疫苗株)和半胱天冬酶抑制剂,建立了一种从克隆cDNA中拯救麻疹病毒的系统。LO-T7-1在CHO/hSLAM细胞中高效驱动T7启动子。基于神经毒性痘苗病毒的LO-T7-1的拯救效率不如vTF7-3株,但通过使用半胱天冬酶抑制剂阻断由LO-T7-1诱导的CHO/hSLAM细胞凋亡,其效率得以提高。这些改进使麻疹病毒的拯救效率甚至高于先前的系统。这个更安全、更高效的系统将进一步促进麻疹病毒在基础研究和病毒载体开发中的基因操作。