State Key Laboratory of Marine Environmental Science, State-Province Joint Engineering Laboratory of Marine Bioproducts and Technology, College of Ocean and Earth Sciences, Xiamen University, Xiamen, 361102, Fujian, China.
State Key Laboratory of Marine Environmental Science, State-Province Joint Engineering Laboratory of Marine Bioproducts and Technology, College of Ocean and Earth Sciences, Xiamen University, Xiamen, 361102, Fujian, China.
Dev Comp Immunol. 2024 Jul;156:105160. doi: 10.1016/j.dci.2024.105160. Epub 2024 Mar 12.
The lacking of stable and susceptible cell lines has hampered research on pathogenic mechanism of crustacean white spot syndrome virus (WSSV). To look for the suitable cell line which can sustain WSSV infection, we performed the studies on WSSV infection in the Spodoptera frugiperda (Sf9) insect cells. In consistent with our previous study in vitro in crayfish hematopoietic tissue cells, the WSSV envelope was detached from nucleocapsid around 2 hpi in Sf9 cells, which was accompanied with the cytoplasmic transport of nucleocapsid toward the cell nucleus within 3 hpi. Furthermore, the expression profile of both gene and protein of WSSV was determined in Sf9 cells after viral infection, in which a viral immediate early gene IE1 and an envelope protein VP28 exhibited gradually increased presence from 3 to 24 hpi. Similarly, the significant increase of WSSV genome replication was found at 3-48 hpi in Sf9 cells after infection with WSSV, indicating that Sf9 cells supported WSSV genome replication. Unfortunately, no assembled progeny virion was observed at 24 and 48 hpi in Sf9 cell nuclei as determined by transmission electron microscope, suggesting that WSSV progeny could not be assembled in Sf9 cell line as the viral structural proteins could not be transported into cell nuclei. Collectively, these findings provide a cell model for comparative analysis of WSSV infection mechanism with crustacean cells.
缺乏稳定和敏感的细胞系一直阻碍着对甲壳动物白斑综合征病毒 (WSSV) 致病机制的研究。为了寻找能持续感染 WSSV 的合适细胞系,我们在草地贪夜蛾 Sf9 昆虫细胞中进行了 WSSV 感染的研究。与我们之前在螯虾造血组织细胞中的体外研究一致,WSSV 包膜在 Sf9 细胞中大约在感染后 2 小时从核衣壳上脱落,伴随着核衣壳在 3 小时内向细胞核内的细胞质转运。此外,在 Sf9 细胞中感染病毒后,确定了 WSSV 的基因和蛋白表达谱,其中病毒的早期基因 IE1 和包膜蛋白 VP28 从 3 小时到 24 小时逐渐增加。同样,在 Sf9 细胞中感染 WSSV 后,在 3-48 小时发现 WSSV 基因组复制显著增加,表明 Sf9 细胞支持 WSSV 基因组复制。然而,遗憾的是,在 Sf9 细胞核中,通过透射电子显微镜观察,在 24 和 48 小时时未观察到组装的子代病毒粒子,表明 WSSV 子代不能在 Sf9 细胞系中组装,因为病毒结构蛋白不能运输到细胞核中。总之,这些发现为比较分析 WSSV 感染机制与甲壳动物细胞提供了一个细胞模型。