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Nanoparticle-based flow virometry for the analysis of individual virions
Anush Arakelyan, … , Leonid Margolis, Jean-Charles Grivel
Anush Arakelyan, … , Leonid Margolis, Jean-Charles Grivel
Published August 8, 2013
Citation Information: J Clin Invest. 2013;123(9):3716-3727. https://doi.org/10.1172/JCI67042.
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Technical Advance AIDS/HIV

Nanoparticle-based flow virometry for the analysis of individual virions

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Abstract

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.

Authors

Anush Arakelyan, Wendy Fitzgerald, Leonid Margolis, Jean-Charles Grivel

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Figure 5

MNP binding and accessibility of cellular antigens on virions.

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MNP binding and accessibility of cellular antigens on virions.
HIV-1SF16...
HIV-1SF162 was produced by PBMCs, captured on MNPs via anti-gp120 antibody VRC01, visualized with labeled 2G12 antibodies, and stained for two cellular antigens with anti–HLA-DR and anti–LFA-1 antibodies. Alternatively, the same preparation of HIV-1SF162 virions was first incubated with anti–LFA-1 and anti–HLA-DR antibodies and then captured on VRC01-MNPs and visualized with 2G12 antibodies. (A) HLA-DR, (B) LFA-1. Left panels: isotype controls. Center panels: staining of MNP-captured virions for HLA-DR and LFA-1. Right panels: staining of virions for HLA-DR and LFA-1 followed by capture with MNPs. Note that the reverse order of staining for cellular antigens and capture with VRC01-MNPs did not significantly affect the result (see the text for the statistical analysis). Results represent one of seven experiments. On each plot, the fraction of events in their respective gates is expressed as a percentage of the total events in the plot.

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