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Whole-blood lymphocyte cultures
E Bloemena1, M T Roos, J L Van Heijst
1Central Laboratory, The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
Journal of Immunological Methods
|September 1, 1989
Summary
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.