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Related Experiment Video

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Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment
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Nonenveloped Avian Reoviruses Released with Small Extracellular Vesicles Are Highly Infectious.

Zuopei Wang1, Menghan He1, Han He2

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Summary

Avian orthoreoviruses (ARVs) can be released within extracellular vesicles (EVs), potentially enhancing their infectivity and transmission. These EVs may contribute to ARV virulence in poultry.

Keywords:
Cryo-EMavian reovirusextracellular vesiclesinfectivitynon-enveloped virus

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Area of Science:

  • Virology
  • Cell Biology
  • Immunology

Background:

  • Avian orthoreoviruses (ARVs) cause significant diseases in poultry.
  • The egress and transmission mechanisms of ARVs, particularly their potential use of extracellular vesicles (EVs), are not well understood.
  • Vesicle-encapsulated viruses can evade immune detection and enhance systemic spread.

Purpose of the Study:

  • To investigate whether ARVs utilize cellular vesicle trafficking for egress and cell-to-cell transmission.
  • To characterize the extracellular vesicles produced by ARV-infected cells.
  • To determine the role of these EVs in ARV infectivity and virulence.

Main Methods:

  • Transmission electron microscopy (TEM) and cryo-electron tomography (Cryo-EM) to visualize vesicles and viral particles.
  • OptiPrep density gradient ultracentrifugation to isolate extracellular vesicles.
  • Western blotting to detect viral proteins within EVs.
  • Infectivity assays to compare the infectivity of virions alone versus virions mixed with EVs.

Main Results:

  • ARV-infected quail cells produced small (~100 nm) extracellular vesicles (EVs).
  • Cryo-EM confirmed that these EVs did not contain intact ARV virions but did contain viral proteins, including the fusogenic p10 FAST protein.
  • EVs containing ARV components significantly increased the infectivity of co-mixed virus particles (up to 50-fold).

Conclusions:

  • ARVs can be released from infected cells within extracellular vesicles.
  • These ARV-associated EVs, potentially carrying fusogenic proteins, may play a role in enhancing viral virulence and transmission.
  • Extracellular vesicles represent a potential mechanism for ARV immune evasion and systemic spread.