Zhang Jun, Chen Xue-Mei, Zhang Chun-Dong, He Qian, Dong Zhan-Qi, Cao Ming-Ya, Dong Xiao-Long, Pan Cai-Xia, Lu Cheng, Pan Min-Hui
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China; Institute of Life Sciences, Chongqing Medical University, Chongqing, China.
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
PLoS One. 2014 Sep 15;9(9):e105986. doi: 10.1371/journal.pone.0105986. eCollection 2014.
We previously established and characterized two insect cell lines (BmN-SWU1 and BmN-SWU2) from Bombyx mori ovaries. Here, we examined their differential susceptibilities to Bombyx mori nucleopolyhedrovirus (BmNPV) despite having originated from the same tissue source. BmN-SWU1 cells were susceptible and supported high titers of BmNPV replication, while BmN-SWU2 cells were resistant to BmNPV infection. Subcellular localization analysis demonstrated that very few BmNPV particles could be imported into BmN-SWU2 cells. However, initiation of BmNPV DNA replication but not amplification was detected in BmN-SWU2 cells after transfection with vA4prm-VP39-EGFP bacmid DNA. BmNPV transcription assays showed that late and very late but not early viral genes apparently were blocked in BmNSWU2 cells by unknown mechanisms. Further syncytium formation assays demonstrated that the BmNPV envelope fusion protein GP64 could not mediate BmN-SWU2 host cell-cell membrane fusion. Taken together, these results indicate that these two cell lines represent optimal tools for investigating host-virus interactions and insect antiviral mechanisms.
我们之前从家蚕卵巢中建立并鉴定了两种昆虫细胞系（BmN-SWU1和BmN-SWU2）。在此，我们研究了它们尽管来源于相同的组织来源，但对家蚕核型多角体病毒（BmNPV）的易感性差异。BmN-SWU1细胞易感并支持高滴度的BmNPV复制，而BmN-SWU2细胞对BmNPV感染具有抗性。亚细胞定位分析表明，很少有BmNPV颗粒能够进入BmN-SWU2细胞。然而，在用vA4prm-VP39-EGFP杆粒DNA转染后，在BmN-SWU2细胞中检测到了BmNPV DNA复制的起始，但未检测到扩增。BmNPV转录分析表明，晚期和极晚期而非早期病毒基因在BmNSWU2细胞中显然被未知机制阻断。进一步的合胞体形成试验表明，BmNPV包膜融合蛋白GP64不能介导BmN-SWU2宿主细胞-细胞膜融合。综上所述，这些结果表明这两种细胞系是研究宿主-病毒相互作用和昆虫抗病毒机制的理想工具。
