Related Experiment Video
Updated: Aug 11, 2026
Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols
Published on: October 18, 2016
Identification of a novel dendritic cell-like subset of CD64(+) / CD16(+) blood monocytes
E Grage-Griebenow1, R Zawatzky, H Kahlert
1Research Center Borstel, Department of Immunology and Cell Biology, Borstel, Germany.
Insights
Researchers identified four human monocyte subsets based on CD64 and CD16 expression. A novel subset combines characteristics of dendritic cells (DCs) and monocytes (Mo), exhibiting high IL-12 production and accessory function.
Area of Science:
- Immunology
- Cell Biology
Background:
- Human monocytes (Mo) comprise distinct subsets with varying functions.
- Fcgamma-receptor I (CD64) and Fcgamma-receptor III (CD16) are key monocyte surface markers.
- Differentiating monocyte subsets is crucial for understanding immune responses.
Purpose of the Study:
- To characterize human monocyte subsets using CD64 and CD16 expression.
- To identify novel monocyte subsets with unique immune functions.
- To elucidate the role of these subsets in antigen presentation and T cell activation.
Main Methods:
- Double labeling of human peripheral blood monocytes with anti-CD64 and anti-CD16 monoclonal antibodies (mAbs).
- Flow cytometry analysis to identify and quantify distinct monocyte subsets.
- Assessment of antigen-presenting cell (APC) function, T cell-accessory capacity, and cytokine production (e.g., IFN-alpha, IL-12).
Main Results:
- Four distinct monocyte subsets were identified: CD64+/16-, CD64-/16+, CD64-/16-, and a novel CD64+/16+ subset.
- The CD64-/16+ subset exhibited high APC function and a macrophage-like phenotype.
- The novel CD64+/16+ subset displayed characteristics of both dendritic cells (DCs) and monocytes, with high IL-12 production and accessory function for lymphocyte activation.
Conclusions:
- Human monocytes can be classified into at least four subsets based on CD64 and CD16 expression.
- A previously undescribed monocyte subset (CD64+/16+) possesses potent immune-modulatory capabilities.
- These findings refine our understanding of monocyte heterogeneity and their roles in adaptive immunity.
Abstract:
Human monocytes (Mo) consist of a major subset of Fcgamma-receptor I (CD64)-positive typical low accessory phagocytes, and a minor CD64(-) DC-like subset with high T cell-accessory and IFN-alpha-releasing activity. Both populations also differentially express CD16 (Fcgamma-receptor III). Double labeling with anti-CD64 and anti-CD16 mAb, as performed here, identified four different subsets. The CD64(-) subset consists of CD64(-) / 16(+) cells with high antigen-presenting cell (APC) function and macrophage-like phenotype, and a CD64(-) / 16(-) subset of less active APC but which exhibits a higher mixed lymphocyte reaction (MLR) stimulating and IFN-alpha-producing capacity, possibly resembling plasmacytoid dendritic cell type II (DC2) blood precursors. As well as the majority of CD64(+) cells that appeared CD64(+) / 16(-) and represent typical low-accessory, CD14(high) Mo, we could identify and describe a novel minor subset of CD64(+) / 16(+) cells which is unique in combining typical DC and Mo characteristics in the same cell. These are high IL-12 production, high accessory capacity for antigen- or allogen-activated lymphocytes, and high expression of HLA-DR, CD86, and CD11c.
