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 (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, safe, and scalable model to study host-virus interactions and the mechanisms of novel therapeutic drugs.
【저자키워드】 COVID-19, SARS-CoV-2, Extracellular vesicles, SARS-CoV-2 spike protein, Colchicine, anti-ACE2, 【초록키워드】 spike, antibody, drugs, clinical trials, Spike protein, cells, Microscopy, therapeutic, host-virus interaction, mechanism, SARS-CoV-2 spike, Analysis, Safe, endothelial, embryonic kidney, coding, internalization, subunits, biogenesis, EVs, Vesicle, Cell, S1 and S2, collected, virus, significantly, generate, reduced, characterized, expressing, bronchial epithelial cell, co-expressed, non-transfected, the SARS-CoV-2, 【제목키워드】 Generation, MIMIC,