Central Facility for Electron Microscopy, Ulm University, 89081, Ulm, Germany.
Bundeswehr Research Institute for Protective Technologies and CBRN Protection, 29633, Munster, Germany.
Histochem Cell Biol. 2021 Dec;156(6):527-538. doi: 10.1007/s00418-021-02028-2. Epub 2021 Sep 12.
Structural changes of two patient-derived glioblastoma cell lines after Zika virus infection were investigated using scanning transmission electron tomography on high-pressure-frozen, freeze-substituted samples. In Zika-virus-infected cells, Golgi structures were barely visible under an electron microscope, and viral factories appeared. The cytosol outside of the viral factories resembled the cytosol of uninfected cells. The viral factories contained largely deranged endoplasmic reticulum (ER), filled with many so-called replication organelles consisting of a luminal vesicle surrounded by the ER membrane. Viral capsids were observed in the vicinity of the replication organelles (cell line #12537 GB) or in ER cisternae at large distance from the replication organelles (cell line #15747 GB). Near the replication organelles, we observed many about 100-nm-long filaments that may represent viral ribonucleoprotein complexes (RNPs), which consist of the RNA genome and N protein oligomers. In addition, we compared Zika-virus-infected cells with cells infected with a phlebovirus (sandfly fever Turkey virus). Zika virions are formed in the ER, whereas phlebovirus virions are assembled in the Golgi apparatus. Our findings will help to understand the replication cycle in the virus factories and the building of the replication organelles in glioblastoma cells.
使用高压冷冻、冷冻替代的方法对 Zika 病毒感染的两株患者来源的神经胶质瘤细胞系进行扫描透射电子显微镜观察，研究其结构变化。在 Zika 病毒感染的细胞中，高尔基氏体在电子显微镜下几乎不可见，出现了病毒工厂。病毒工厂外的细胞质类似于未感染细胞的细胞质。病毒工厂内含有大量排列紊乱的内质网（ER），充满了许多所谓的复制细胞器，由一个被 ER 膜包围的腔室样囊泡组成。在复制细胞器附近（细胞系#12537GB）或远离复制细胞器的 ER 腔中（细胞系#15747GB）观察到病毒衣壳。在复制细胞器附近，我们观察到许多大约 100nm 长的纤维，它们可能代表病毒核糖核蛋白复合物（RNP），由 RNA 基因组和 N 蛋白寡聚体组成。此外，我们还将 Zika 病毒感染的细胞与感染沙蝇热病毒（phlebovirus）的细胞进行了比较。Zika 病毒在 ER 中形成，而 phlebovirus 在高尔基氏体中组装。我们的研究结果将有助于理解病毒工厂中的复制周期和神经胶质瘤细胞中复制细胞器的构建。
