Abstract
Extracellular vesicles (EVs) are secreted membrane vesicles, derived from endosomes or from the plasma membrane, which have been isolated from most cell types and biological fluids. Although EVs are highly heterogeneous and their classification is complex, two major categories can be distinguished: microvesicles (MVs), which derive from the shedding of the plasma membrane, and exosomes, which correspond to intraluminal vesicles released to the extracellular milieu upon fusion of multivesicular bodies (MVBs) with the plasma membrane. Cells infected with viruses may secrete MVs containing viral proteins, RNAs and, in some instances, infectious virions. A recent study carried out by our laboratory has shown that MVs released by cells infected with HSV-1 contained virions and were endocytosed by naïve cells leading to a productive infection. This suggests that HSV-1 may use MVs for spreading, expanding its tropism and evading the host immune response. Here we describe in detail the methods used to isolate and analyse the MVs released from HSV-1-infected cells.
Key words
- HSV-1
- Extracellular vesicles
- Microvesicles
- Differential centrifugation protocols
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Acknowledgments
We thank María Teresa Rejas, Technical Director of the Electron Microscopy Facility (CBMSO), for her support with the electron-microscopy protocols.
Funding: Fundación Severo Ochoa-Aeromédica Canaria provided financial support. The funders had no role in study design, decision to publish, or preparation of the manuscript.
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Bello-Morales, R., López-Guerrero, J.A. (2020). Isolation/Analysis of Extracellular Microvesicles from HSV-1-Infected Cells. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus . Methods in Molecular Biology, vol 2060. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9814-2_17
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