Detection of the interactions of tumour derived extracellular vesicles with immune cells is dependent on EV‐labelling methods

Detection of the interactions of tumour derived extracellular vesicles with immune cells is dependent on EV‐labelling methods

Cell‐cell communication within the complex tumour microenvironment is critical to cancer progression. Tumor‐derived extracellular vesicles (TD‐EVs) are key players in this process. They can interact with immune cells and modulate their activity, either suppressing or activating the immune system. De...

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Published in:Journal of extracellular vesicles Vol. 12; no. 12; pp. e12384 - n/a
Main Authors: Loconte, Luisa, Arguedas, Davinia, El, Rojbin, Zhou, Alix, Chipont, Anna, Guyonnet, Lea, Guerin, Coralie, Piovesana, Ester, Vázquez‐Ibar, José Luis, Joliot, Alain, Théry, Clotilde, Martín‐Jaular, Lorena
Format: Journal Article
Language:English
Published: United States John Wiley & Sons, Inc 01-12-2023
Taylor & Francis
John Wiley and Sons Inc
Wiley
Subjects:
Breast carcinoma
Carboxyfluorescein diacetate
CD14 antigen
Cell interactions
Cell Line
Dendritic cells
Dyes
Ectopic expression
Extracellular vesicles
Extracellular Vesicles - metabolism
Flow cytometry
Humans
immune cells
Immune system
Immunomodulation
Labeling
labelling
Leukocytes, Mononuclear
Life Sciences
Lipids
Lipophilic
Monocytes
Nanoparticles
Peripheral blood mononuclear cells
Plasmids
Proteins
Software
Succinimides - metabolism
Technical Note
Tumor microenvironment
Tumors
uptake
immune cells
extracellular vesicles
labelling
uptake
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Summary:Cell‐cell communication within the complex tumour microenvironment is critical to cancer progression. Tumor‐derived extracellular vesicles (TD‐EVs) are key players in this process. They can interact with immune cells and modulate their activity, either suppressing or activating the immune system. Deciphering the interactions between TD‐EVs and immune cells is essential to understand immune modulation by cancer cells. Fluorescent labelling of TD‐EVs is a method of choice to study such interaction. This work aims to determine the impact of EV labelling methods on the detection by imaging flow cytometry and multicolour spectral flow cytometry of EV interaction and capture by the different immune cell types within human Peripheral Blood Mononuclear Cells (PBMCs). EVs released by the triple‐negative breast carcinoma cell line MDA‐MB‐231 were labelled either with the lipophilic dye MemGlow‐488 (MG‐488), Carboxyfluorescein diacetate, succinimidyl ester (CFDA‐SE) or through ectopic expression of a MyrPalm‐superFolderGFP reporter (mp‐sfGFP), which incorporates into EVs during their biogenesis. Our results show that these labelling strategies, although analysed with the same techniques, led to diverging results. While MG‐488‐labelled EVs incorporate in all cell types, CFSE‐labelled EVs are restricted to a minor subset of cells and mp‐sfGFP‐labelled EVs are mainly detected in CD14+ monocytes which are the main uptakers of EVs and other particles, regardless of the labelling method. Furthermore, our results show that the method used for EV labelling influences the detection of the different types of EV interactions with the recipient cells. Specifically, MG‐488, CFSE and mp‐sfGFP result in observation suggesting, respectively, transient EV‐PM interaction that results in dye transfer, EV content delivery, and capture of intact EVs. Consequently, the type of EV labelling method has to be considered as they can provide complementary information on various types of EV‐cell interaction and EV fate.
Bibliography:Ester Piovesana, Laboratory for Aging Disorders, Laboratories for Translational Research, EOC Bellinzona (Bios+), Switzerland.
Davinia Arguedas and Rojbin El contributed equally to this study.
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ISSN:2001-3078
2001-3078
DOI:10.1002/jev2.12384