Nanoparticle-based flow virometry for the analysis of individual virions
Nanoparticle-based flow virometry for the analysis of individual virions
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DOI：
10.1172/jci67042
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发表时间：
2013-09-01
影响因子：
15.9
通讯作者：
Grivel, Jean-Charles
中科院分区：
文献类型：
--
作者：
Arakelyan, Anush;Fitzgerald, Wendy;Grivel, Jean-Charles
虽然流式细胞术已用于分析单个细胞的抗原组成，但病毒颗粒的抗原组成仍主要以散装形式表征。在这里，我们描述了一种技术，“流动病毒测定法”，可用于单个病毒体的抗原检测。该技术基于将磁性纳米颗粒结合到病毒体，用单克隆抗体染色病毒体，用磁性柱分离形成的复合物，并用流式细胞仪对其进行表征。我们使用这种技术来研究两种抗原（HLA-DR和LFA-1）的分布，HIV-1从感染的细胞中获得单个HIV-1病毒粒子。流式病毒测定法显示，来自单一制剂的病毒体的抗原组成是异质的。这种异质性无法通过病毒的批量分析检测到。此外，在由不同细胞产生的相同HIV-1的两种制备物中，抗原在病毒体中的分布是不同的。相比之下，在同一细胞中复制的两种不同HIV-1基因型的HIV-1在抗原性上变得有些相似。这种纳米技术允许研究体液中的病毒粒子而不需要病毒繁殖，原则上不限于HIV的分析，而是可以应用于任何病毒的个体表面抗原组成的分析。
While flow cytometry has been used to analyze the antigenic composition of individual cells, the antigenic makeup of viral particles is still characterized predominantly in bulk. Here, we describe a technology, "flow virometry," that can be used for antigen detection on individual virions. The technology is based on binding magnetic nanoparticles to virions, staining the virions with monoclonal antibodies, separating the formed complexes with magnetic columns, and characterizing them with flow cytometers. We used this technology to study the distribution of two antigens (HLA-DR and LFA-1) that HIV-1 acquires from infected cells among individual HIV-1 virions. Flow virometry revealed that the antigenic makeup of virions from a single preparation is heterogeneous. This heterogeneity could not be detected with bulk analysis of viruses. Moreover, in two preparations of the same HIV-1 produced by different cells, the distribution of antigens among virions was different. In contrast, HIV-1 of two different HIV-1 genotypes replicating in the same cells became somewhat antigenically similar. This nanotechnology allows the study of virions in bodily fluids without virus propagation and in principle is not restricted to the analysis of HIV, but can be applied to the analysis of the individual surface antigenic makeup of any virus.