Human Blood Monocyte Subsets
Top Cited Papers
Open Access
- 1 August 2017
- journal article
- Published by Ovid Technologies (Wolters Kluwer Health) in Arteriosclerosis, Thrombosis, and Vascular Biology
- Vol. 37 (8), 1548-1558
- https://doi.org/10.1161/atvbaha.117.309145
Abstract
Objective—: Human monocyte subsets are defined as classical (CD14 ++ CD16 − ), intermediate (CD14 ++ CD16 + ), and nonclassical (CD14 + CD16 + ). Alterations in monocyte subset frequencies are associated with clinical outcomes, including cardiovascular disease, in which circulating intermediate monocytes independently predict cardiovascular events. However, delineating mechanisms of monocyte function is hampered by inconsistent results among studies. Approach and Results—: We use cytometry by time-of-flight mass cytometry to profile human monocytes using a panel of 36 cell surface markers. Using the dimensionality reduction approach visual interactive stochastic neighbor embedding (viSNE), we define monocytes by incorporating all cell surface markers simultaneously. Using viSNE, we find that although classical monocytes are defined with high purity using CD14 and CD16, intermediate and nonclassical monocytes defined using CD14 and CD16 alone are frequently contaminated, with average intermediate and nonclassical monocyte purity of ≈86.0% and 87.2%, respectively. To improve the monocyte purity, we devised a new gating scheme that takes advantage of the shared coexpression of cell surface markers on each subset. In addition to CD14 and CD16, CCR2, CD36, HLA-DR, and CD11c are the most informative markers that discriminate among the 3 monocyte populations. Using these additional markers as filters, our revised gating scheme increases the purity of both intermediate and nonclassical monocyte subsets to 98.8% and 99.1%, respectively. We demonstrate the use of this new gating scheme using conventional flow cytometry of peripheral blood mononuclear cells from subjects with cardiovascular disease. Conclusions—: Using cytometry by time-of-flight mass cytometry, we have identified a small panel of surface markers that can significantly improve monocyte subset identification and purity in flow cytometry. Such a revised gating scheme will be useful for clinical studies of monocyte function in human cardiovascular disease.Keywords
This publication has 24 references indexed in Scilit:
- Comparison of two different strategies for human monocyte subsets gating within the large‐scale prospective CARE FOR HOMe StudyCytometry Part A, 2015
- High-dimensional analysis of the murine myeloid cell systemNature Immunology, 2014
- Toward a Refined Definition of Monocyte SubsetsFrontiers in Immunology, 2013
- Monocyte heterogeneity in human cardiovascular diseaseImmunobiology, 2012
- The three human monocyte subsets: implications for health and diseaseImmunologic Research, 2012
- Gene expression profiling reveals the defining features of the classical, intermediate, and nonclassical human monocyte subsetsBlood, 2011
- Differential regulation of toll-like receptor-2, toll-like receptor-4, CD16 and human leucocyte antigen-DR on peripheral blood monocytes during mild and severe dengue feverImmunology, 2010
- Increased subpopulations of CD16+ and CD56+ blood monocytes in patients with active Crohnʼs diseaseInflammatory Bowel Diseases, 2007
- Expression of CD14, CD16 and CD45RA on monocytes from periodontitis patientsJournal of Periodontal Research, 2003
- A Global Geometric Framework for Nonlinear Dimensionality ReductionScience, 2000