ACE2-containing defensosomes serve as decoys to inhibit SARS-CoV-2 infection

Extracellular vesicles of endosomal origin, exosomes, mediate intercellular communication by transporting substrates with a variety of functions related to tissue homeostasis and disease. Their diagnostic and therapeutic potential has been recognized for diseases such as cancer in which signaling defects are prominent. However, it is unclear to what extent exosomes and their cargo inform the progression of infectious diseases.

New York University researchers recently defined a subset of exosomes termed defensosomes that are mobilized during bacterial infection in a manner dependent on autophagy proteins. Through incorporating protein receptors on their surface, defensosomes mediated host defense by binding and inhibiting pore-forming toxins secreted by bacterial pathogens. Given this capacity to serve as decoys that interfere with surface protein interactions, the researchers investigated the role of defensosomes during infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of COVID-19. Consistent with a protective function, exosomes containing high levels of the viral receptor ACE2 in bronchioalveolar lavage fluid from critically ill COVID-19 patients was associated with reduced ICU and hospitalization times. They found ACE2+ exosomes were induced by SARS-CoV-2 infection and activation of viral sensors in cell culture, which required the autophagy protein ATG16L1, defining these as defensosomes. The researchers further demonstrate that ACE2+ defensosomes directly bind and block viral entry. These findings suggest that defensosomes may contribute to the antiviral response against SARS-CoV-2 and expand our knowledge on the regulation and effects of extracellular vesicles during infection.

ACE2+ exosomes protect against SARS-CoV-2 infection

(A) Workflow for exosome neutralization assay used to assess protective effects of mixing SARS-CoV-2 and exosomes. Created with BioRender.com. (B) Representative histogram of cell-surface ACE2 on A549 and A549ACE2 cells. (C) Representative immunofluorescence images of Vero E6 cells infected with SARS-CoV-2, pellet from 5000xg spin, ~20K exosomes from ACE2- cells, ~20K exosomes from ACE2+ cells, neutralizing antibody, or with media alone stained for viral N protein. (D) Percentage of infected Vero E6 cells based on positive staining for N protein after infection with SARS-CoV-2 mixed with exosomes isolated from A549ACE2 (teal) or untransduced A549 cells (black). (E) Representative immunofluorescence images of HAECs infected with mock (media only control), SARS-CoV-2 alone, ~11K (1) or ~22K (2) ACE2+ exosomes with virus, ~150K (3) ACE2- exosomes with virus, and neutralizing antibody with virus (nAb). Images were taken 72h.p.i. (F) Representative flow cytometry plots of exosomes isolated from Vero E6 cells stimulated with CpG. FSC: forward scatter, SSC: side scatter. (G) Quantification of exosomes from Vero E6 cells and Vero E6 cells transduced with a non-targeting shRNA for ATG16L1 stimulated with PBS (n=4) or 1uM CpG-A (n=4) for 16 hr. (H) Quantification of infected cells after infection of Vero E6 cells with SARS-CoV-2 only, SARS-CoV-2 mixed with supernatants of cells pretreated overnight with CpG-A (induced), or SARS-CoV-2 added to CpG-A pretreated cells following removal of supernatant (wash).