Gaudin Raphaël, Barteneva Natasha S
1] Program in Cellular and Molecular Medicine, Boston Children's Hospital, 200 Longwood Avenue, Boston, Massachusetts 02115, USA [2] Department of Cell Biology, Harvard Medical School, 200 Longwood Avenue, Boston, Massachusetts 02115, USA.
1] Program in Cellular and Molecular Medicine, Boston Children's Hospital, 200 Longwood Avenue, Boston, Massachusetts 02115, USA [2] Department of Pediatrics, Harvard Medical School, 200 Longwood Avenue, Boston, Massachusetts 02115, USA.
Nat Commun. 2015 Feb 2;6:6022. doi: 10.1038/ncomms7022.
The nature and concentration of lipids and proteins at the surface of viruses are essential parameters for determining particle infectiveness. Historically, averaged bulk analysis of viral particles has been the primary method to quantitatively investigate these parameters, though this neglects heterogeneity within populations. Here we analyse the properties of Junin virus particles using a sensitive flow virometry assay and further sort virions while conserving their infectiveness. This method allows us to characterize the relationship between infectivity, virus size and RNA content and to compare particles secreted by Vero cells with those from physiologically relevant human primary macrophages. Our study highlights significant differences in particle infectivity according to its nature, the type of producer cells and the lipid membrane composition at the budding site. Together, our results present the flow virometry assay as a powerful and versatile tool to define virus particle profiles.
病毒表面脂质和蛋白质的性质及浓度是决定颗粒感染性的关键参数。从历史上看，对病毒颗粒进行平均整体分析一直是定量研究这些参数的主要方法，不过这种方法忽略了群体内部的异质性。在此，我们使用灵敏的流式病毒测定法分析胡宁病毒颗粒的特性，并在保持病毒粒子感染性的同时对其进行进一步分选。该方法使我们能够表征感染性、病毒大小与RNA含量之间的关系，并比较由Vero细胞分泌的颗粒与来自生理相关的人类原代巨噬细胞的颗粒。我们的研究突出了根据病毒颗粒的性质、产生细胞的类型以及出芽位点的脂膜组成，颗粒感染性存在显著差异。总之，我们的结果表明流式病毒测定法是定义病毒颗粒概况的一种强大且通用的工具。
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