4.5 Article

PBMC-derived extracellular vesicles in a smoking-related inflammatory disease model

Journal

EUROPEAN JOURNAL OF IMMUNOLOGY
Volume 53, Issue 6, Pages -

Publisher

WILEY
DOI: 10.1002/eji.202250143

Keywords

Extracellular vesicles; Exovesicles; Imaging flow cytometry; Spectral flow cytometry

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This study aimed to optimize methodologies to study immune cell-derived extracellular vesicles (EVs). Peripheral blood mononuclear cell-derived small EVs were isolated and characterized using conventional transmission electron microscopy and nanoparticle tracking analysis. They were further analyzed using imaging flow cytometry and flow cytometric multiplex bead assay. These techniques were applied to smoking-related inflammatory disease research following the Minimal Information for Studies of Extracellular Vesicle 2018 guidelines.
Extracellular vesicles (EVs) function as mediators of intercellular communication and as such influence the recipient cell function. EVs derived from immune cells can carry out many of the same functions as their parental cells, as they carry costimulatory molecules, antigens, and antigen-MHC complexes. As a result, there is a strong interest in understanding the composition and origin of immune cell-derived EVs in order to understand their role in the pathogenesis of diseases. This study aimed to optimize methodologies to study immune cell-derived EVs. Peripheral blood mononuclear cell-derived small EVs were isolated and observed using conventional transmission electron microscopy and sized by nanoparticle tracking analysis. They were then enumerated and profiled using imaging flow cytometry and were further characterized using a flow cytometric multiplex bead assay. These techniques were then applied to our current research, namely smoking-related inflammatory disease. We present here a comprehensive approach to analyze PBMC-derived small EVs in smoking-related inflammatory disease following the Minimal Information for Studies of Extracellular Vesicle 2018 guidelines.

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