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Updated: Aug 11, 2026
Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro
Published on: June 2, 2010
Whole-blood lymphocyte cultures
E Bloemena1, M T Roos, J L Van Heijst
1Central Laboratory, The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
Insights
This study presents a simple, reproducible method to measure lymphocyte proliferation using whole blood, eliminating the need for cell separation. This technique offers reliable results for studying immune responses, even in immunocompromised patients.
Area of Science:
- Immunology
- Cellular Biology
Background:
- Assessing lymphocyte proliferation is crucial for understanding immune function.
- Traditional methods often require complex cell separation, increasing variability and labor.
Purpose of the Study:
- To develop a simple and reproducible whole blood assay for measuring lymphocyte proliferative responses.
- To establish reference values for this novel assay.
- To demonstrate its utility in longitudinal studies of immunocompromised individuals.
Main Methods:
- Utilized small blood aliquots without prior cell separation.
- Stimulated whole blood lymphocytes with horse anti-human lymphocyte serum (ALS), phytohaemagglutinin (PHA), or anti-CD3 monoclonal antibody (mAb).
- Measured proliferative responses expressed as counts per minute (cpm) per volume or per CD3+ lymphocyte.
Main Results:
- The method demonstrated high reproducibility and low variation.
- Reference values for proliferation were established using percentile distributions.
- The assay proved effective for longitudinal monitoring in immunocompromised subjects.
Conclusions:
- This whole blood lymphocyte proliferation assay is a robust and efficient tool for immunological studies.
- It simplifies sample processing and enhances reliability.
- The assay is suitable for clinical applications, including monitoring immunocompromised patients.
Abstract:
A simple and reproducible method is described for the measurement of proliferative responses of peripheral blood lymphocytes in whole blood upon stimulation with horse anti-human lymphocyte serum (ALS), phytohaemagglutinin (PHA) or a monoclonal antibody (mAb) directed against CD3. Only small aliquots of blood are needed and separation procedures are not necessary. Since the method proved to be very reproducible and showed little variation, reference values of proliferation, expressed in cpm per 15 microliters blood and cpm per 10(3) CD3+ lymphocytes, were determined based upon the 5th and the 2.5th percentile of the distribution of the values in our reference population. In addition, the use of whole blood lymphocyte cultures in longitudinal studies of immunocompromised individuals is demonstrated.
