TY - JOUR
T1 - Sorting of small infectious virus particles by flow virometry reveals distinct infectivity profiles
AU - Gaudin, Raphaël
AU - Barteneva, Natasha S.
N1 - Funding Information:
We are thankful to Larry Duckett, David Matsuyama and Paul Melanson, BD Biosciences Inc. (San Jose, CA) for their help with the cytometer’s upgrade and functionality. We are grateful to Dr Michael W. Olszowy (Life Technologies) for providing the 40-and 100-nm fluorescent beads. We thank Lloyd Bahlmann for giving us access to Izon technology. We also thank Tom Kirchhausen for letting us use his laboratory and microscopes and Ivan Vorobjev for his helpful advice. We thank Frances Evesson for her comments and critical reading of the manuscript. We greatly acknowledge the Harvard Medical School Electron Microscopy facility. N.S.B. was supported by the Harvard Pilot Grant and PCMM/Boston Children’s Hospital. R.G. was supported by the National Institutes of Health Grants GM-075252 and U54 AI057159 (New England Regional Center of Excellence in Biodefense and Emerging Infectious Diseases [NERCE BEID]).
PY - 2015/2
Y1 - 2015/2
N2 - 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.
AB - 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.
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U2 - 10.1038/ncomms7022
DO - 10.1038/ncomms7022
M3 - Article
C2 - 25641385
AN - SCOPUS:84923098091
VL - 6
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
M1 - 6022
ER -