Journal article
Comparison of different methods for isolating CD8+ T lymphocyte-derived extracellular vesicles and supramolecular attack particles
- Abstract:
- CD8+ T lymphocytes play vital roles in killing infected or deranged host cells, recruiting innate immune cells, and regulating other aspects of immune responses. Like any other cell, CD8+ T cells also produce extracellular particles. These include extracellular vesicles (EVs) and non-vesicular extracellular particles (NVEPs). T cell-derived EVs are proposed to mediate cell-to-cell signalling, especially in the context of inflammatory responses, autoimmunity, and infectious diseases. CD8+ T cells also produce supramolecular attack particles (SMAPs), which are in the same size range as EVs and mediate a component of T cell mediated killing. The isolation technique selected will have a profound effect on yield, purity, biochemical properties and function of T cell-derived particles; making it important to directly compare different approaches. In this study, we compared commonly used techniques (membrane spin filtration, ultracentrifugation, or size exclusion liquid chromatography) to isolate particles from activated human CD8+ T cells and validated our results by single-particle methods, including nanoparticle tracking analysis, flow cytometry, electron microscopy and super-resolution microscopy of the purified sample as well as bulk proteomics and lipidomics analyses to evaluate the quality and nature of enriched T cell-derived particles. Our results show that there is a trade-off between the yield and the quality of T cell-derived particles. Furthermore, the protein and lipid composition of the particles is dramatically impacted by the isolation technique applied. We conclude that from the techniques evaluated, size exclusion liquid chromatography offers the highest quality of T cell derived EVs and SMAPs with acceptable yields for compositional and functional studies.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 3.9MB, Terms of use)
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- Publisher copy:
- 10.1002/jex2.74
Authors
+ University of Oxford
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- Funder identifier:
- https://ror.org/052gg0110
- Funding agency for:
- Jainarayanan, AK
- Grant:
- BB/M011224/1
- Programme:
- Interdisciplinary Bioscience DTP
+ European Commission
More from this funder
- Funder identifier:
- https://ror.org/00k4n6c32
- Grant:
- ERC-2021-SyG_95132
- ERC-2014-AdG_670930
- Programme:
- Horizon 2020
+ Wellcome Trust
More from this funder
- Funder identifier:
- https://ror.org/029chgv08
- Grant:
- 100262Z/12/Z
- 224343/Z/21/Z
+ Kennedy Trust for Rheumatology Research
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- Grant:
- KENN202117
- Programme:
- Cell Dynamics Program
- Publisher:
- Wiley
- Journal:
- Journal of Extracellular Biology More from this journal
- Volume:
- 2
- Issue:
- 3
- Article number:
- e74
- Publication date:
- 2023-03-13
- Acceptance date:
- 2023-01-30
- DOI:
- EISSN:
-
2768-2811
- Language:
-
English
- Keywords:
- Pubs id:
-
1327501
- Local pid:
-
pubs:1327501
- Deposit date:
-
2023-02-06
- ARK identifier:
Terms of use
- Copyright holder:
- Jainarayanan et al.
- Copyright date:
- 2023
- Rights statement:
- © 2023 The Authors. Journal of Extracellular Biology published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Notes:
- A correction to this article was published on 2 May 2023 at https://doi.org/10.1002/jex2.83
- Licence:
- CC Attribution (CC BY)
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