Institute of Virology, Ulm University Medical Center, 89081 Ulm, Germany.
Int J Mol Sci. 2022 Oct 8;23(19):11931. doi: 10.3390/ijms231911931.
PML nuclear bodies (PML-NBs) are dynamic macromolecular complexes that mediate intrinsic immunity against viruses of different families, including human cytomegalovirus (HCMV). Upon HCMV infection, PML-NBs target viral genomes entering the nucleus and restrict viral immediate-early gene expression by epigenetic silencing. Studies from several groups performed in human fibroblast cells have shown that the major PML-NB components PML, Daxx, Sp100 and ATRX contribute to this repression in a cooperative manner. Their role for HCMV restriction in endothelial cells, however, has not yet been characterized although infected endothelium is thought to play a crucial role for HCMV dissemination and development of vascular disease in vivo. Here, we use conditionally immortalized umbilical vein endothelial cells (HEC-LTT) as a cell culture model to elucidate the impact of PML-NB proteins on lytic HCMV infection. Depletion of individual PML-NB proteins by lentiviral transduction showed a particularly strong antiviral effect of PML in HEC-LTT, compared to human fibroblasts. A closer characterization of this antiviral function revealed that PML may not only effectively inhibit HCMV immediate-early gene expression but also act at later steps of the viral replication cycle. At contrast, we surprisingly noted an antiviral behavior of Daxx in complementary approaches: Depletion of Daxx resulted in decreased viral gene expression, while overexpression of Daxx promoted HCMV infection. In summary, our data demonstrate a cell type-specific effect of PML-NB components on lytic HCMV infection and suggest an important role of PML in the inhibition of HCMV dissemination through infected endothelial cells.
PML 核小体(PML-NBs)是一种动态的大分子复合物,可介导针对不同家族病毒(包括人巨细胞病毒(HCMV))的固有免疫。在 HCMV 感染后,PML-NBs 靶向进入细胞核的病毒基因组,并通过表观遗传沉默来限制病毒即刻早期基因的表达。来自多个小组在人成纤维细胞中的研究表明,主要的 PML-NB 成分 PML、Daxx、Sp100 和 ATRX 以协作的方式共同促进这种抑制作用。然而,它们在血管内皮细胞中对 HCMV 的限制作用尚未得到表征,尽管受感染的内皮细胞被认为在体内 HCMV 的传播和血管疾病的发展中起着至关重要的作用。在这里,我们使用条件永生化脐静脉内皮细胞(HEC-LTT)作为细胞培养模型,阐明 PML-NB 蛋白对裂解型 HCMV 感染的影响。通过慢病毒转导耗尽单个 PML-NB 蛋白显示,与人类成纤维细胞相比,PML 在 HEC-LTT 中对 HCMV 的抗病毒作用特别强。对这种抗病毒功能的更深入分析表明,PML 不仅可以有效地抑制 HCMV 即刻早期基因的表达,而且还可以作用于病毒复制周期的后期步骤。相比之下,我们在互补方法中令人惊讶地注意到 Daxx 的抗病毒行为:Daxx 的耗竭导致病毒基因表达减少,而过表达 Daxx 则促进 HCMV 感染。总之,我们的数据表明 PML-NB 成分对裂解型 HCMV 感染具有细胞类型特异性的影响,并表明 PML 在通过受感染的内皮细胞抑制 HCMV 传播方面发挥重要作用。