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Updated: Aug 13, 2026
Enumeration of Major Peripheral Blood Leukocyte Populations for Multicenter Clinical Trials Using a Whole Blood Phenotyping Assay
Published on: September 16, 2012
[Lymphocyte subsets in blood donors]
L Larrea1, R García Boyero, A Sempere
1Servicio de Hematología Hospital Universitario La Fe, Valencia.
Insights
This study found no significant differences in lymphocyte subsets (T, B, NK cells) or absolute counts based on age, gender, or sample origin in healthy blood donors. Reference ranges were established for laboratory use.
Area of Science:
- Immunology
- Hematology
Context:
- Establishing normal reference ranges for lymphocyte subsets is crucial for accurate clinical interpretation.
- Flow cytometry is a standard method for enumerating lymphocyte populations.
- Variations in lymphocyte counts can be influenced by demographic factors.
Purpose:
- To determine normal values for lymphocyte subsets (T, B, NK cells) in healthy individuals.
- To investigate potential demographic influences (age, gender, origin) on lymphocyte subset distribution.
- To compare absolute lymphocyte counts obtained via flow cytometry and automated cell counters.
Summary:
- A prospective study of 100 blood donors analyzed lymphocyte subsets using direct immunofluorescence and flow cytometry.
- No significant differences were observed in lymphocyte subset counts concerning age, gender, or sample origin.
- Absolute lymphocyte counts from flow cytometry and automated cell counters showed no significant discrepancies.
Impact:
- Provides established reference ranges for lymphocyte subsets in a healthy population.
- Demonstrates the reliability of flow cytometry for lymphocyte enumeration compared to automated counters.
- Highlights the lack of significant demographic influence on key lymphocyte populations.
Objectives:
To establish the normal values among the different lymphocyte subsets in peripheral blood, measured by surface antigen expression, in healthy population. To observe if there are potential differences in respect of age, gender or sample origin. To compare the absolute lymphocyte number obtained with flow cytometry and with a cellular counter.
Material And Methods:
Longitudinal and prospective study performed with 100 samples of blood donors. Direct immunofluorescence with triple color staining was made on whole blood, red blood cells were then lysed and samples were analysed with a flow cytometer. The lymphocyte subsets studied were the T lymphocytes and their subsets (CD4 and CD8 lymphocytes), the B lymphocytes and Natural Killer cells population. The absolute lymphocyte count was performed with an automatic cellular counter. Donor data such as age, gender and origin were recorded and were statistically analysed.
Results:
Normal ranges from the studied parameters are similar to other series. Comparison with gender, origin or age groups gave no significant difference, although there seems to be a tendency to decrease with the ageing of the population. Total absolute lymphocyte number did not differ between the results from the cellular counter or from the flow cytometer.
Conclusions:
We found no differences in the absolute lymphocyte number nor in the lymphocyte subsets studied (T lymphocytes, CD4 lymphocytes, CD8 lymphocytes, B lymphocytes and NK cells) with respect to gender, age or sample source. We have established reference ranges for our laboratory. We have not found significant differences in the absolute lymphocyte number measured with cell counter or with flow cytometer.
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