Department of Nephrology, University of Heidelberg, Heidelberg, Germany.
Department of Medicine, Rush University, Medical Center, Chicago, IL, USA.
Virus Res. 2019 Jan 15;260:102-113. doi: 10.1016/j.virusres.2018.11.015. Epub 2018 Nov 30.
Orthohantaviruses Hantaan (HTNV) and Puumala (PUUV) virus cause hemorrhagic fever with renal syndrome (HFRS), that is characterized by acute renal failure with often massive proteinuria and by morphological changes of the tubular and glomerular apparatus. Orthohantaviral N protein is found in renal cells and plays a key role in replication. However, the replication in human renal cells is not well characterized. Therefore, we examined the orthohantaviral infection in different human renal cells. Differences in localization of N protein, release of particles, and modulation of the actin cytoskeleton between both virus species are observed in human renal cells. A substantial portion of HTNV N protein demonstrates a filamentous pattern in addition to the typical punctate pattern. Release of HTNV depends on an intact actin and microtubule cytoskeleton. In contrast, PUUV N protein is generally localized in a punctate pattern and release of PUUV does not require an intact actin cytoskeleton. Infection of podocytes results in cytoskeletal rearrangements that are more pronounced for HTNV. Analyzing Vero E6 cells revealed differences compared to human renal cells. The pattern of N proteins is strictly punctate, release does not depend on an intact actin cytoskeleton and cytoskeletal rearrangements are not present. No virus-specific variations between HTNV and PUUV are observed in Vero E6 cells. Using human renal cells as cell culture model for orthohantavirus infection demonstrates virus-specific differences and orthohantavirus-induced cytoskeletal rearrangements that are not observed in Vero E6 cells. Therefore, the choice of an appropriate cell culture system is a prerequisite to study orthohantavirus pathogenicity.
汉坦病毒（HTNV）和普马拉病毒（PUUV）引起肾综合征出血热（HFRS），其特征为急性肾衰竭常伴有大量蛋白尿和肾小管及肾小球装置的形态学改变。正粘病毒 N 蛋白存在于肾细胞中，在复制中起关键作用。然而，人肾细胞中的复制情况尚未得到很好的描述。因此，我们检查了不同人肾细胞中的正粘病毒感染。在人肾细胞中观察到 N 蛋白的定位、颗粒释放和肌动蛋白细胞骨架的调节在两种病毒之间存在差异。HTNV N 蛋白的很大一部分除了典型的点状模式外，还呈现出丝状模式。HTNV 的释放依赖于完整的肌动蛋白和微管细胞骨架。相比之下，PUUV N 蛋白通常定位于点状模式，PUUV 的释放不需要完整的肌动蛋白细胞骨架。足细胞感染导致细胞骨架重排，HTNV 更为明显。分析 Vero E6 细胞与人类肾细胞相比存在差异。N 蛋白的模式严格是点状的，释放不依赖于完整的肌动蛋白细胞骨架，也不存在细胞骨架重排。在 Vero E6 细胞中，未观察到 HTNV 和 PUUV 之间的病毒特异性差异。使用人肾细胞作为正粘病毒感染的细胞培养模型，证明了在 Vero E6 细胞中未观察到的病毒特异性差异和正粘病毒诱导的细胞骨架重排。因此，选择合适的细胞培养系统是研究正粘病毒致病性的前提。
