Quantitative characterisation of extracellular vesicles designed to decoy or compete with SARS-CoV-2 reveals differential mode of action across variants of concern and highlights the diversity of Omicron

Publication date: Jul 03, 2025

Blue: DAPI; Green: J2 anti-dsRNA; Red (left panel): anti- SARS-CoV2-nucleocapsid protein; Red (right panel): anti-SARS-CoV-2 Spike protein; Purple: anti-acetylated tubulin (Cilia). b Particles per mg protein ratios for the different engineered and native HEK293T EVs, as assessed by normalizing particle concentrations determined by NTA to protein concentrations determined by Bradford. To simultaneously monitor uptake of the S-GFP sp EV subpopulation over all CD63 positive EVs, we used EVs from cells double transfected with either CD63-mCherry and S-GFP sp (Fig. f Western Blot of cell lysates derived from native and S-GFP sp transfected HEK293T cells and on recombinant SARS-CoV2 Spike protein with an anti-S-Protein antibody (GeneTex). Assay performed on CHO ACE2 +/T2 + cells or on CHO ACE2 -/T2- cells using PV engineered to display different Spike protein mutants (SARS-COV-2: D614G, BA. 1 Omicron; SARS-COV-1). c Single vesicle imaging performed on CMs collected from HEK293T cells transfected with different SGFP sp plasmids. Particles detected by the antibody are pseudo coloured in red, fluorescent protein (FP)tagged EVs (GFP, tGFP, GFP sp or mNeon) are shown in green and co-localisation in yellow.

Semantics

Original Article