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Isolation and Characterization of Neutrophil-derived Microparticles for Functional Studies
Published on: March 2, 2018
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Human Neutrophils Generate Extracellular Vesicles That Modulate Their Functional Responses
María José Hurtado Gutiérrez1, Frédérick L Allard1, Hugo Tshivuadi Mosha1
1Department of Immunology and Cell Biology, Medicine Faculty, Université de Sherbrooke, CRCHUS, Sherbrooke, QC J1H5N4, Canada.
Cells
|January 8, 2023
Summary
Neutrophil-derived ectosomes (NDEs) modulate immune cell function. These extracellular vesicles (EVs) influence autologous neutrophil responses, impacting survival, chemokine release, and neutrophil extracellular trap (NET) formation.
Area of Science:
- Immunology
- Cell Biology
- Extracellular Vesicles
Background:
- Neutrophils are key immune cells influencing both innate and adaptive immunity.
- Neutrophils release extracellular vesicles (EVs), specifically ectosomes, which can affect other immune cells.
- The autocrine and paracrine functions of neutrophil-derived ectosomes (NDEs) on neutrophil responses require further elucidation.
Purpose of the Study:
- To investigate the role of neutrophil-derived ectosomes (NDEs) in modulating autologous neutrophil responses.
- To characterize the production and release of EVs by neutrophils according to MISEV 2018 guidelines.
- To determine how NDEs influence neutrophil survival, chemokine production, and neutrophil extracellular trap (NET) formation.
Main Methods:
- Characterization of neutrophil-derived EVs (ectosomes) following MISEV 2018 guidelines.
- Stimulation of neutrophils with various physiological agents to induce EV release.
- Co-incubation of NDEs with autologous neutrophils to assess effects on cellular responses.
- Analysis of neutrophil survival, cytokine/chemokine production (e.g., CXCL8), and NET formation.
Main Results:
- Neutrophils primarily release NDEs, which are rapidly released upon certain physiological stimuli.
- NDE constituents are incorporated into autologous neutrophils, modulating their function.
- NDEs reversed the pro-survival effects of GM-CSF, G-CSF, IFNγ, and dexamethasone.
- NDEs induced CXCL8 and NET formation in unstimulated neutrophils and modulated TNFα-induced chemokine release.
- NDEs attenuated inducible NET generation.
Conclusions:
- Neutrophil-derived ectosomes (NDEs) play a significant role in modulating autologous neutrophil responses through autocrine and paracrine mechanisms.
- NDE generation is an important mechanism by which neutrophils shape their microenvironment and cellular interactions.
- These findings highlight NDEs as critical mediators in neutrophil-mediated immunity.
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