• Media type: E-Article
  • Title: Generation of Spike-Extracellular Vesicles (S-EVs) as a Tool to Mimic SARS-CoV-2 Interaction with Host Cells
  • Contributor: Verta, Roberta; Grange, Cristina; Skovronova, Renata; Tanzi, Adele; Peruzzi, Licia; Deregibus, Maria Chiara; Camussi, Giovanni; Bussolati, Benedetta
  • imprint: MDPI AG, 2022
  • Published in: Cells
  • Language: English
  • DOI: 10.3390/cells11010146
  • ISSN: 2073-4409
  • Keywords: General Medicine
  • Origination:
  • Footnote:
  • Description: <jats:p>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.</jats:p>
  • Access State: Open Access