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Updated: May 30, 2026
Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis
Published on: October 17, 2018
SuperSAGE evidence for CD14++CD16+ monocytes as a third monocyte subset
Adam M Zawada1, Kyrill S Rogacev, Björn Rotter
1Department of Internal Medicine IV, Saarland University Hospital, Homburg/Saar, Germany.
Insights
This study characterizes CD14++CD16+ monocytes, revealing unique genetic identifiers and diverse immune functions including antigen processing, inflammation, and angiogenesis. These findings highlight a distinct role for this monocyte subset in human immunity.
Area of Science:
- Immunology
- Cell Biology
- Genomics
Background:
- Monocytes are key immune cells with diverse subsets, including classical (CD14++CD16-), intermediate (CD14++CD16+), and nonclassical (CD14+CD16++) populations.
- Human monocyte heterogeneity is not fully understood, particularly the CD14++CD16+ subset, despite its known significance in HIV-1 infection and atherosclerosis.
Purpose of the Study:
- To develop a method for purifying human monocyte subsets.
- To analyze the transcriptomes of purified monocyte subsets using SuperSAGE and high-throughput sequencing.
- To identify unique genetic identifiers and functions of CD14++CD16+ monocytes.
Main Methods:
- Purification of three human monocyte subsets (CD14++CD16-, CD14++CD16+, CD14+CD16++) from blood.
- Transcriptome analysis using SuperSAGE (Suppression Subtractive Hybridization and RACE) and high-throughput sequencing.
- Gene Ontology (GO) enrichment analysis to determine cellular functions.
Main Results:
- Successfully purified distinct monocyte subsets.
- Identified unique transcriptomic identifiers for CD14++CD16+ monocytes.
- GO analysis linked CD14++CD16+ monocytes to antigen processing/presentation, inflammation, monocyte activation, and angiogenesis.
Conclusions:
- Provided genetic evidence for a distinct role of CD14++CD16+ monocytes in human immunity.
- The findings offer insights into the poorly characterized CD14++CD16+ monocyte subset.
- This research may stimulate further investigation into monocyte heterogeneity and potential therapeutic targets.
Abstract:
Monocytes are a heterogeneous cell population with subset-specific functions and phenotypes. The differential expression of CD14 and CD16 distinguishes classical CD14(++)CD16(-), intermediate CD14(++)CD16(+), and nonclassical CD14(+)CD16(++) monocytes. Current knowledge on human monocyte heterogeneity is still incomplete: while it is increasingly acknowledged that CD14(++)CD16(+) monocytes are of outstanding significance in 2 global health issues, namely HIV-1 infection and atherosclerosis, CD14(++)CD16(+) monocytes remain the most poorly characterized subset so far. We therefore developed a method to purify the 3 monocyte subsets from human blood and analyzed their transcriptomes using SuperSAGE in combination with high-throughput sequencing. Analysis of 5 487 603 tags revealed unique identifiers of CD14(++)CD16(+) monocytes, delineating these cells from the 2 other monocyte subsets. Gene Ontology (GO) enrichment analysis suggests diverse immunologic functions, linking CD14(++)CD16(+) monocytes to Ag processing and presentation (eg, CD74, HLA-DR, IFI30, CTSB), to inflammation and monocyte activation (eg, TGFB1, AIF1, PTPN6), and to angiogenesis (eg, TIE2, CD105). In conclusion, we provide genetic evidence for a distinct role of CD14(++)CD16(+) monocytes in human immunity. After CD14(++)CD16(+) monocytes have earlier been discussed as a potential therapeutic target in inflammatory diseases, we are hopeful that our data will spur further research in the field of monocyte heterogeneity.
