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Updated: Dec 26, 2025
Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis
Published on: October 17, 2018
Expression of CD163 and HLA-DR molecules on the monocytes in chronic lymphocytic leukemia patients
1Chair and Department of Clinical Immunology Medical University of Lublin, Lublin, Poland. w.kowalska.lub@gmail.com.
Insights
Monocyte subsets in chronic lymphocytic leukemia (CLL) patients show distinct characteristics. CD16-positive monocytes are elevated, with classical monocytes expressing CD163 and non-classical monocytes showing low HLA-DR, potentially impacting immune responses.
Area of Science:
- Immunology
- Hematology
- Oncology
Background:
- Human peripheral blood monocytes play a role in the leukemia microenvironment.
- Three monocyte subpopulations exist: classical (CD14++CD16-), intermediate (CD14++CD16+), and non-classical (CD14+CD16++).
- These subpopulations differ in HLA-DR and CD163 expression.
Purpose of the Study:
- To evaluate HLA-DR and CD163 expression on monocyte subpopulations in patients with chronic lymphocytic leukemia (CLL).
- To compare monocyte subpopulations between CLL patients and healthy individuals.
Main Methods:
- Flow cytometry was used to analyze peripheral blood monocyte subsets.
- Monocytes from 40 CLL patients and 10 healthy controls were analyzed.
- Specific monoclonal antibodies against CD14, CD16, CD163, and HLA-DR were employed.
Main Results:
- CD16-positive monocytes were significantly higher in CLL patients compared to healthy donors.
- Classical monocytes (CD14++CD16-) showed the highest percentage of CD163 expression.
- Non-classical monocytes (CD14+CD16++) were the predominant cells with low HLA-DR expression.
- No significant correlation was found between monocyte subpopulation percentages and CLL stage (Rai Staging).
Conclusions:
- CD163 expression on classical monocytes suggests potential anti-inflammatory properties.
- Low HLA-DR expression on non-classical monocytes may indicate impaired immune stimulation capacity in CLL.
- Monocyte subset analysis provides insights into the immune microenvironment in CLL.
Introduction:
Human peripheral blood monocytes are the part of the leukemia microenvironment. We examined three monocyte subgroups: classical (CD14++CD16-), intermediate (CD14++CD16+) and non-classical (CD14+CD16++) monocytes. As these subpopulations can be also characterized by different levels of HLA-DR and CD163, we evaluated their expression on monocyte subpopulations of patients with chronic lymphocytic leukemia (CLL) and healthy individuals.
Material And Methods:
The monocyte subsets in peripheral blood of CLL patients (n = 40) and healthy controls (n = 10) were evaluated by flow cytometry. The monoclonal antibodies: anti-CD14 FITC, anti-CD16 PE-Cy5, anti-CD163 PE, anti-HLA-DR PE were used.
Results:
The percentage of CD16-positive monocytes was significantly higher in CLL patients than in healthy donors. The highest percentage of CD163+ monocytes is in the 'classical' (CD14++CD16-) population. In turn, the non-classical monocytes constituted the majority of cells lacking HLA-DR expression. In CLL patients, there was no statistically significant relationship between the percentage of each monocyte subpopulation and the stage according to Rai Staging of CLL.
Conclusions:
The presence of CD163 on classical monocytes suggests that these cells have anti-inflammatory properties. Besides, the low expression of HLA-DR on non-classical monocytes may result in impaired ability to stimulate the immune system.
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