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In enteric viral infections, such as those with rotavirus and norovirus, individual viral particles shed in stool are considered the optimal units of fecal-oral transmission. We reveal that rotaviruses and noroviruses are also shed in stool as viral clusters enclosed within vesicles that deliver a high inoculum to the receiving host. Cultured cells non-lytically release rotaviruses and noroviruses inside extracellular vesicles. In addition, stools of infected hosts contain norovirus and rotavirus within vesicles of exosomal or plasma membrane origin. These vesicles remain intact during fecal-oral transmission and thereby transport multiple viral particles collectively to the next host, enhancing both the MOI and disease severity. Vesicle-cloaked viruses are non-negligible populations in stool and have a disproportionately larger contribution to infectivity than free viruses. Our findings indicate that vesicle-cloaked viruses are highly virulent units of fecal-oral transmission and highlight a need for antivirals targeting vesicles and virus clustering.
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Fig 6. (A and B) Vesicles previously isolated from EDIM rotavirus-infected mouse stool samples were stained with CellBrite Fix 488 fluorescent membrane dye and fed to mouse pups. Intestinal contents 30 min post feeding were observed under the confocal microscope live (A) or fixed and immunostained with anti-rotavirus VP6 antibody (B). Representative images from two independent experiments are presented. Scale bars, 2.5 μm (A) and 1 μm (B). (Picture credits: doi:10.1016/j.chom.2018.07.006)
In this study, the CellBrite™ Steady, Membrane Staining Kits was used to demonstrate that multiple viruses were present inside membrane enclosed vesicles of plasma membrane or exosomal origin, depending on the viral strain. Labeling with CellBrite™ Fix 488 was also used to demonstrate uptake of the viral vesicles through the endocytic pathway, and to track their passage through the GI tract . Learn more about the CellBrite™ Steady Membrane Staining Kits.
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Farmer BC, Kluemper J, Johnson LA. Apolipoprotein E4 Alters Astrocyte Fatty Acid Metabolism and Lipid Droplet Formation. Cells. 2019 Feb 20;8(2). pii: E182. doi: 10.3390/cells8020182.
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