title: Evaluation of the maturation of individual Dengue virions with flow virometry.
abstract: High-throughput techniques are needed to analyze individual virions to understand how viral heterogeneity translates into pathogenesis since in bulk analysis the individual characteristics of virions are lost. Individual Dengue virions (DENV) undergo a maturation that involves a proteolytic cleavage of prM precursor into virion-associated M protein. Here, using a new nanoparticle-based technology, "flow virometry", we compared the maturation of individual DENV produced by BHK-21 and LoVo cells. The latter lacks the furin-protease that mediates prM cleavage. We found that prM is present on about 50% of DENV particles produced in BHK-21 cells and about 85% of DENV virions produced in LoVo, indicating an increase in the fraction of not fully matured virions. Flow virometry allows us to quantify the number of fully mature particles in DENV preparations and proves to be a useful method for studying heterogeneity of the surface proteins of various viruses.
authors.affiliation: Section on Intercellular, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Sonia
authors.initials: S
authors.lastname: Zicari
authors.affiliation: Section on Intercellular, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Anush
authors.initials: A
authors.lastname: Arakelyan
authors.affiliation: Section on Intercellular, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Wendy
authors.initials: W
authors.lastname: Fitzgerald
authors.affiliation: Section on Membrane Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, United States.
authors.forename: Elena
authors.initials: E
authors.lastname: Zaitseva
authors.affiliation: Section on Membrane Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, United States. Electronic address: chernoml@mail.nih.gov.
authors.forename: Leonid V
authors.initials: LV
authors.lastname: Chernomordik
authors.affiliation: Section on Intercellular, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, United States. Electronic address: margolil@helix.nih.gov.
authors.forename: Leonid
authors.initials: L
authors.lastname: Margolis
authors.affiliation: Sidra Medical and Research Center, Doha, Qatar.
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Grivel
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003716:Dengue Virus / Q000502:physiology*; D006801:Humans; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014771:Virion / Q000378:metabolism*; D014773:Virology / Q000379:methods*
chemical_list: D014759:Viral Envelope Proteins; C074932:prM protein, Flavivirus
keywords: Dengue; Flow virometry; Maturation; prM