An intra-cytoplasmic route for SARS-CoV-2 transmission unveiled by Helium-ion microscopy

Sci Rep. 2022 Mar 8;12(1):3794. doi: 10.1038/s41598-022-07867-0.

Abstract

SARS-CoV-2 virions enter the host cells by docking their spike glycoproteins to the membrane-bound Angiotensin Converting Enzyme 2. After intracellular assembly, the newly formed virions are released from the infected cells to propagate the infection, using the extra-cytoplasmic ACE2 docking mechanism. However, the molecular events underpinning SARS-CoV-2 transmission between host cells are not fully understood. Here, we report the findings of a scanning Helium-ion microscopy study performed on Vero E6 cells infected with mNeonGreen-expressing SARS-CoV-2. Our data reveal, with unprecedented resolution, the presence of: (1) long tunneling nanotubes that connect two or more host cells over submillimeter distances; (2) large scale multiple cell fusion events (syncytia); and (3) abundant extracellular vesicles of various sizes. Taken together, these ultrastructural features describe a novel intra-cytoplasmic connection among SARS-CoV-2 infected cells that may act as an alternative route of viral transmission, disengaged from the well-known extra-cytoplasmic ACE2 docking mechanism. Such route may explain the elusiveness of SARS-CoV-2 to survive from the immune surveillance of the infected host.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Angiotensin-Converting Enzyme 2 / metabolism
  • Animals
  • COVID-19 / transmission
  • COVID-19 / virology
  • Chlorocebus aethiops
  • Cytoplasm / chemistry
  • Cytoplasm / ultrastructure
  • Cytoplasm / virology
  • Extracellular Vesicles / chemistry
  • Extracellular Vesicles / ultrastructure
  • Giant Cells / chemistry
  • Giant Cells / physiology
  • Helium / chemistry
  • Humans
  • Ions / chemistry
  • Microscopy / methods*
  • SARS-CoV-2 / isolation & purification
  • SARS-CoV-2 / physiology*
  • Spike Glycoprotein, Coronavirus / chemistry
  • Spike Glycoprotein, Coronavirus / metabolism
  • Vero Cells
  • Virus Internalization*

Substances

  • Ions
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2
  • Helium
  • Angiotensin-Converting Enzyme 2