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Published on: February 8, 2016
Immune cell phenotyping in low blood volumes for assessment of cardiovascular disease risk, development, and
Yvonne Baumer1, Cristhian A Gutierrez-Huerta1, Ankit Saxena2
1Social Determinants of Obesity and Cardiovascular Risk Laboratory, National Heart Lung and Blood Institute, National Institutes of Health, Building 10-CRC, Room 5-5332, Bethesda, MD, 20892, USA.
Insights
This study developed a flow cytometry panel to analyze immune cells in cardiovascular disease (CVD) risk. The panel revealed distinct immune cell profiles between African-Americans and Caucasians, highlighting potential new CVD risk factors.
Area of Science:
- Immunology
- Cardiovascular Science
- Flow Cytometry
Background:
- Cardiovascular disease (CVD) is a leading global cause of mortality.
- Immune cell involvement in atherosclerosis necessitates advanced characterization methods, especially for at-risk populations.
Purpose of the Study:
- To develop and validate a flow cytometry panel for detailed immune cell sub-population analysis.
- To compare immune cell profiles between African-Americans (AA) and Caucasians.
- To investigate the association between immune cell characteristics and CVD risk factors.
Main Methods:
- A multi-antibody flow cytometry panel was designed to identify granulocyte, lymphocyte, and monocyte subsets.
- The panel was tested on healthy individuals (N=29) stratified by race/ethnicity (AA and Caucasian).
- Immune cell parameters were analyzed in a clinical cohort of AA women with CVD risk factors (N=20).
Main Results:
- The flow cytometry panel successfully differentiated immune cell sub-populations, revealing differences between AA and Caucasians (e.g., granulocyte and lymphocyte proportions).
- Novel differences were observed, including increased platelet adhesion on non-classical monocytes in AA.
- Flow cytometry parameters, such as immune cell platelet aggregates, correlated with clinical CVD risk markers like the Framingham Risk Score (FRS).
Conclusions:
- The developed flow cytometry panel effectively identifies immune cell differences between racial groups, potentially contributing to CVD risk disparities.
- This method can phenotype immune cell sub-populations and platelet aggregates linked to CVD risk.
- The panel shows promise as a tool for understanding immune cell roles in CVD development and progression.
Background:
Cardiovascular disease (CVD) is the leading cause of death in the world. Given the role of immune cells in atherosclerosis development and progression, effective methods for characterizing immune cell populations are needed, particularly among populations disproportionately at risk for CVD.
Results:
By using a variety of antibodies combined in one staining protocol, we were able to identify granulocyte, lymphocyte, and monocyte sub-populations by CD-antigen expression from 500 µl of whole blood, enabling a more extensive comparison than what is possible with a complete blood count and differential (CBC). The flow cytometry panel was established and tested in a total of 29 healthy men and women. As a proof of principle, these 29 samples were split by their race/ethnicity: African-Americans (AA) (N = 14) and Caucasians (N = 15). We found in accordance with the literature that AA had fewer granulocytes and more lymphocytes when compared to Caucasians, though the proportion of total monocytes was similar in both groups. Several new differences between AA and Caucasians were noted that had not been previously described. For example, AA had a greater proportion of platelet adhesion on non-classical monocytes when compared to Caucasians, a cell-to-cell interaction described as crucially important in CVD. We also examined our flow panel in a clinical population of AA women with known CVD risk factors (N = 20). Several of the flow cytometry parameters that cannot be measured with the CBC displayed correlations with clinical CVD risk markers. For instance, Framingham Risk Score (FRS) calculated for each participant correlated with immune cell platelet aggregates (PA) (e.g. T cell PA β = 0.59, p = 0.03 or non-classical monocyte PA β = 0.54, p = 0.02) after adjustment for body mass index (BMI).
Conclusion:
A flow cytometry panel identified differences in granulocytes, monocytes, and lymphocytes between AA and Caucasians which may contribute to increased CVD risk in AA. Moreover, this flow panel identifies immune cell sub-populations and platelet aggregates associated with CVD risk. This flow cytometry panel may serve as an effective method for phenotyping immune cell populations involved in the development and progression of CVD.
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