Immunophenotypic characterization of human monocyte subsets: possible implications for cardiovascular disease pathophysiology

Background: Monocytes are critical to the initiation and development of atherosclerosis. To date, 3 distinct human monocyte subsets have been identified based primarily on their expression of the surface markers CD14 and CD16. With the emerging knowledge of myeloid-derived suppressor cells and other myeloid subsets, we hypothesized that monocytes are likely more heterogeneous in composition. Therefore, we set out to use the high dimensionality of mass cytometry to accurately identify and define monocyte subsets in blood of healthy humans and their changes in cardiovascular patients. Methods: Heparinized blood from 12 healthy donors and 15 patients with defined cardiovascular disease (CVD) based on angiography and gensini score was obtained and analyzed by CyTOF mass cytometry. We employed the Phenograph algorithm to cluster and identify all healthy monocyte subsets based on their phenotypes using a 40-marker mass cytometry panel. Results: Phenograph identified a total of 15 monocyte clusters in healthy human blood. By performing hierarchical clustering, we were able to group these clusters into 6 larger meta-clusters and found that most of these meta-clusters fall within the CD14 classical monocyte population, illustrating significant heterogeneity among this monocyte population. Cell numbers of one of these monocyte meta-clusters were significantly increased in blood from patients with CVD. We also identified two subsets of nonclassical monocytes in healthy donors. One of these subsets showed higher expression of the integrin CD61 and tetraspanin CD9, pointing to a possible role for this subset in patrolling and platelet activation. Conclusion: Monocytes are highly diverse with the conventional classical subset showing the most diversity. The numbers and frequencies of some of these monocyte subsets are changed in CVD. Studies to identify their functions in CVD should provide new information for the role of monocytes in CVD.

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Isolation and Characterization of Human Monocyte Subsets

Macrophages and Natural Killer Cells - Advances in Experimental Medicine and Biology ◽

10.1007/978-1-4684-4394-3_20 ◽

1982 ◽

pp. 209-219 ◽

Cited By ~ 3

Author(s):

Roy S. Weiner

Keyword(s):

Human Monocyte ◽

Monocyte Subsets ◽

Isolation And Characterization

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Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis

Journal of Visualized Experiments ◽

10.3791/57941 ◽

2018 ◽

Cited By ~ 9

Author(s):

Rekha Marimuthu ◽

Habib Francis ◽

Suat Dervish ◽

Stephen C.H. Li ◽

Heather Medbury ◽

...

Keyword(s):

Flow Cytometry ◽

Blood Flow ◽

Whole Blood ◽

Human Monocyte ◽

Flow Cytometry Analysis ◽

Monocyte Subsets

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Role of CCR8 and Other Chemokine Pathways in the Migration of Monocyte-derived Dendritic Cells to Lymph Nodes

Journal of Experimental Medicine ◽

10.1084/jem.20032152 ◽

2004 ◽

Vol 200 (10) ◽

pp. 1231-1241 ◽

Cited By ~ 220

Author(s):

Chunfeng Qu ◽

Emmerson W. Edwards ◽

Frank Tacke ◽

Véronique Angeli ◽

Jaime Llodrá ◽

...

Keyword(s):

Dendritic Cells ◽

Lymph Nodes ◽

Chemokine Receptors ◽

Human Monocyte ◽

Monocyte Subsets ◽

Migration Pathways

Studying the influence of chemokine receptors (CCRs) on monocyte fate may reveal information about which subpopulations of monocytes convert to dendritic cells (DCs) and the migration pathways that they use. First, we examined whether prominent CCRs on different monocyte subsets, CCR2 or CX3CR1, mediated migration events upstream of the accumulation of monocyte-derived DCs in lymph nodes (LNs). Monocytes were labeled and traced by uptake of latex microspheres in skin. Unexpectedly, neither CCR2 nor CX3CR1 were required. However, absence of CCR2 led to an increased labeling of the minor Gr-1int monocyte population, and the number of latex+ DCs that emigrated to LNs was correspondingly increased. Characterization of Gr-1int monocytes revealed that they selectively expressed CCR7 and CCR8 mRNA in blood. CCR7 and CCR8 pathways were used by monocyte-derived DCs during mobilization from skin to LNs. The role of CCR8 in emigration from tissues also applied to human monocyte-derived cells in a model of transendothelial trafficking. Collectively, the data suggest that Gr-1int monocytes may be most disposed to become a lymphatic-migrating DCs. When these monocyte-derived DCs exit skin to emigrate to LNs, they use not only CCR7 but also CCR8, which was not previously recognized to participate in migration to LNs.

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Molecular Characterization of Monocyte Subsets Reveals Specific and Distinctive Molecular Signatures Associated With Cardiovascular Disease in Rheumatoid Arthritis

Frontiers in Immunology ◽

10.3389/fimmu.2019.01111 ◽

2019 ◽

Vol 10 ◽

Cited By ~ 4

Author(s):

Patricia Ruiz-Limon ◽

Rafaela Ortega-Castro ◽

Nuria Barbarroja ◽

Carlos Perez-Sanchez ◽

Christophe Jamin ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Cardiovascular Disease ◽

Molecular Characterization ◽

Molecular Signatures ◽

Monocyte Subsets

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IL-33 stimulates the release of procoagulant microvesicles from human monocytes and differentially increases tissue factor in human monocyte subsets

Thrombosis and Haemostasis ◽

10.1160/th16-10-0784 ◽

2017 ◽

Vol 117 (07) ◽

pp. 1379-1390 ◽

Cited By ~ 16

Author(s):

Stefan Stojkovic ◽

Åsa Thulin ◽

Lena Hell ◽

Barbara Thaler ◽

Sabine Rauscher ◽

...

Keyword(s):

Cardiovascular Disease ◽

Tissue Factor ◽

Inflammatory Diseases ◽

Human Monocyte ◽

Receptor Expression ◽

Human Monocytes ◽

Monocyte Subsets ◽

Prothrombotic State ◽

St2 Receptor ◽

Classical Monocytes

SummaryMonocytes and monocyte-derived microvesicles (MVs) are the main source of circulating tissue factor (TF). Increased monocyte TF expression and increased circulating levels of procoagulant MVs contribute to the formation of a prothrombotic state in patients with cardiovascular disease. Interleukin (IL)-33 is a pro-inflammatory cytokine involved in atherosclerosis and other inflammatory diseases, but its role in regulating thrombosis is still unclear. The aim of the present study was to investigate the effects of IL-33 on the procoagulant properties of human monocytes and monocyte-derived MVs. IL-33 induced a time- and concentration-dependent increase of monocyte TF mRNA and protein levels via binding to the ST2-receptor and activation of the NFκB-pathway. The IL-33 treated monocytes also released CD14+TF+ MVs and IL-33 was found to increase the TF activity of both the isolated monocytes and monocyte-derived MVs. The monocytes were classified into subsets according to their CD14 and CD16 expression. Intermediate monocytes (IM) showed the highest ST2 receptor expression, followed by non-classical monocytes (NCM), and classical monocytes (CM). IL-33 induced a significant increase of TF only in the IM (p<0.01), with a tendency in NCM (p=0.06), but no increase was observed in CM. Finally, plasma levels of IL–33 were positively correlated with CD14+TF+ MVs in patients undergoing carotid endarterectomy (r=0.480; p=0.032; n=20). We hereby provide novel evidence that the proinflammatory cytokine IL-33 induces differential TF expression and activity in monocyte subsets, as well as the release of procoagulant MVs. In this manner, IL-33 may contribute to the formation of a prothrombotic state characteristic for cardiovascular disease.Supplementary Material to this article is available online at www.thrombosis-online.com.

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