Generation of Spike-Extracellular Vesicles (S-EVs) as a Tool to Mimic SARS-CoV-2 Interaction with Host Cells

Generation of Spike-Extracellular Vesicles (S-EVs) as a Tool to Mimic SARS-CoV-2 Interaction with Host Cells

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Extracellular vesicles (EVs) and viruses share common features: size, structure, biogenesis and uptake.In order to generate EVs expressing the SARS-CoV-2 spike protein on their surface ethical nutrients mega magnesium powder citrus 450g (S-EVs), we collected EVs from SARS-CoV-2 spike expressing human embryonic kidney (HEK-293T) cells by stable transfection with a vector coding for the S1 and S2 subunits.S-EVs were characterized using nanoparticle tracking analysis, ExoView and super-resolution microscopy.We obtained a population of EVs of 50 to 200 nm in size.

Spike expressing EVs represented around 40% of the total EV population and co-expressed spike protein with tetraspanins on the surfaces of EVs.We subsequently used ACE2-positive endothelial and bronchial epithelial cells for assessing the internalization of labeled S-EVs using a cytofluorimetric analysis.Internalization of S-EVs was higher than that of control EVs from non-transfected cells.Moreover, S-EV uptake was significantly decreased by anti-ACE2 antibody pre-treatment.

Furthermore, colchicine, a drug currently used in clinical trials, significantly reduced S-EV entry into the cells.S-EVs represent a simple, il barone wine safe, and scalable model to study host-virus interactions and the mechanisms of novel therapeutic drugs.