Department of Entomology Institute of Plant Protection, Agricultural Research Organization, Rishon Lezion 7528809, Israel.
Viruses. 2020 Jul 9;12(7):739. doi: 10.3390/v12070739.
Many attempts to develop a reliable cell cultured-based system to study honey bee virus infections have encountered substantial difficulties. We investigated the ability of a cell line from a heterologous insect to sustain infection by a honey bee virus. For this purpose, we infected the Lepidopteran hemocytic cell line (P1) with Deformed wing virus (DWV). The genomic copies of DWV increased upon infection, as monitored by quantitative RT-PCR. Moreover, a tagged-primer-based RT-PCR analysis showed the presence of DWV negative-sense RNA in the cells, indicating virus replication. However, the DWV from infected cells was mildly infectious to P1 cells. Similar results were obtained when the virus was injected into pupae. Thus, though the virus yields from the infected cells appeared to be very low, we show for the first time that DWV can replicate in a heterologous cell line. Given the availability of many other insect cell lines, our study paves the way for future exploration in this direction. In the absence of adequate cell lines, exploring the ability of alternative cell lines to enable honey bee virus infections could provide the means to study and understand the viral infectious cycle at the cellular level and facilitate obtaining purified isolates of these viruses.
许多尝试开发可靠的基于细胞培养的系统来研究蜜蜂病毒感染的尝试都遇到了很大的困难。我们研究了一种来自异源昆虫的细胞系维持蜜蜂病毒感染的能力。为此,我们用变形翅膀病毒(DWV)感染鳞翅目血细胞系(P1)。通过定量 RT-PCR 监测,感染后 DWV 的基因组拷贝数增加。此外,基于标记引物的 RT-PCR 分析表明细胞中存在 DWV 负链 RNA,表明病毒复制。然而,来自受感染细胞的 DWV 对 P1 细胞的感染性较弱。当将病毒注射到蛹中时,得到了类似的结果。因此,尽管从感染细胞中获得的病毒产量似乎非常低,但我们首次表明 DWV 可以在异源细胞系中复制。鉴于有许多其他昆虫细胞系可用,我们的研究为未来在这一方向的探索铺平了道路。在缺乏足够的细胞系的情况下,探索替代细胞系是否能够使蜜蜂病毒感染,可以提供在细胞水平上研究和理解病毒感染周期的手段,并有助于获得这些病毒的纯化分离物。