期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 517, 期 4, 页码 709-714出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2019.07.120
关键词
Monocyte isolation; Monocyte subsets; Extracellular vesicles; Flow cytometry
资金
- Christian Doppler Society (Christian Doppler Laboratory for Innovative Therapy Approaches in Sepsis)
- regional government of Lower Austria
- EFRE
Human monocytes include CD14(++)CD16(-) (classical), CD14(++)CD16(+)(intermediate), and CD14(+)CD16(++)(non-classical) subsets with divergent roles in immune regulation and inflammation. Since the functional characterization of monocyte subsets is most commonly performed using isolated monocytes, we investigated the influence of different monocyte isolation protocols on the relative abundance of monocyte subsets. Using flow cytometric subset characterization directly in whole blood as a reference, we found that monocyte isolation by enrichment of peripheral blood mononuclear cells and subsequent depletion of non-monocytes by magnetic labeling did not alter the distribution of monocyte subsets. Particularly, we failed to detect a loss of CD16(+) subsets upon monocyte isolation, although one of the negative depletion protocols used contained an anti-CD16 antibody to label granulocytes. Overnight storage of isolated monocytes induced a significant repartition of monocyte subsets towards CD14(++)CD16(+) intermediate monocytes, which was barely seen in stored whole blood. We identified intermediate monocytes as main binding partners of platelet-derived extracellular vesicles (EVs) and propose that residual platelets contained in isolated monocyte preparations release EVs that induce the expression of the IgG receptor Fc gamma RIII (CD16) on monocytes. (C) 2019 The Authors. Published by Elsevier Inc.
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