College of Life Science, Central China Normal University, Wuhan, 430079, People's Republic of China.
Cytotechnology. 2008 Jul;57(3):233-43. doi: 10.1007/s10616-008-9158-8. Epub 2008 Aug 20.
It is well known that the characteristics of cell lines possibly alter when cell lines are at high-passage number because of the environmental selection. We do not know whether non-permissive or low-permissive cell lines could become permissive or more permissive to virus infection after over-high passage. In the present studies, the alteration of the permissiveness of Spodoptera litura cell line Sl-zsu-1 to three baculovirus infection was investigated after over-high passage, and the possible mechanisms are also investigated. Vigorous apoptosis in Sl-zsu-1 cells was induced by both the recombinant Autographa californica multiple nucleopolyhedrovirus AcMNPV-GFP-actin and the celery looper Anagrapha falcifera multiple nucleopolyhedrovirus AfMNPV, suggesting the replication of the two viruses was blocked by apoptosis. However, the cells infected by S. litura multicapsid nucleopolyhedrovirus SpltMNPV did not undergo apoptosis, but the SpltMNPV titre of the supernatant was not detectable, suggesting this cell line was low-permissive for this virus infection and other factor(s) involved in blockage of the virus replication except apoptosis. However, when Sl-zsu-1 cells had been subcultured continuously for more than 4 years (high-passage cell), which was named as Sl-HP cell line afterwards, no significant apoptosis was induced by the three baculovirus in Sl-HP cells, and many replicated virions or nucleocapsids were observed in the cells. But the permissiveness of Sl-HP cells to the three viruses was very different according to the titre of viruses in the cell cultures. Interestingly, the DNA extracted from SpltMNPV could induce vigorous apoptosis of Sl-HP cells. Altogether, Sl-zsu-1 cell line becomes more permissive to baculovirus infection after over-high passage and multiple paths can block the baculovirus infectivity.
众所周知,由于环境选择,细胞系在高传代数时其特征可能会发生改变。我们不知道经过超高传代后,非允许或低允许细胞系是否能够对病毒感染变得允许或更允许。在本研究中,研究了超高传代后斜纹夜蛾细胞系 Sl-zsu-1 对三种杆状病毒感染的允许性改变,并探讨了可能的机制。重组杆状病毒 AcMNPV-GFP-actin 和芹菜夜蛾杆状病毒 AfMNPV 均可诱导 Sl-zsu-1 细胞发生强烈的细胞凋亡,表明这两种病毒的复制被细胞凋亡所阻断。然而,感染 SpltMNPV 的细胞不发生细胞凋亡,但上清液中的 SpltMNPV 滴度不可检测,表明该细胞系对该病毒感染低允许,除了细胞凋亡之外,还有其他因素参与病毒复制的阻断。然而,当 Sl-zsu-1 细胞连续传代培养超过 4 年(高传代细胞)后,命名为 Sl-HP 细胞系,在 Sl-HP 细胞中,三种杆状病毒均不能诱导明显的细胞凋亡,并且可以观察到大量复制的病毒粒子或核衣壳。但是,根据细胞培养物中病毒的滴度,Sl-HP 细胞对三种病毒的允许性差异很大。有趣的是,从 SpltMNPV 提取的 DNA 可以诱导 Sl-HP 细胞发生强烈的细胞凋亡。总之,超高传代后斜纹夜蛾细胞系对杆状病毒感染的允许性增加,并且有多种途径可以阻断杆状病毒的感染力。