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Measuring Lymphocyte Proliferation in Response to Specific Antigen and Mitogen Stimuli Using Flow Cytometry
Clinical Laboratory
|February 7, 2017
Summary
This study introduces a flow cytometry system for precise in-vitro immunofunction testing. Individual immune responses to stimuli are stable over time, suggesting a potential genetic basis for immune reactivity.
Area of Science:
- Immunology
- Cellular Biology
- Biotechnology
Background:
- Developing accurate methods to assess in-vitro immunofunction is crucial.
- Existing methods like 3H-thymidine assays have limitations in precision and phenotyping.
Purpose of the Study:
- To establish a simple, CFSE-based multivariate flow cytometry system for analyzing lymphocyte proliferation.
- To quantify stimulus-induced lymphocyte proliferation dynamics with high precision.
Main Methods:
- Utilizing multicolour flow cytometry for lymphocyte stimulation with 5 mitogens and 7 antigens.
- Employing CFSE (carboxyfluorescein succinimidyl ester) dye to track cell division.
- Quantifying proliferation by resolving daughter generations and determining precursor frequency.
Main Results:
- The developed system offers greater precision in measuring lymphocyte proliferation compared to radionucleotide methods.
- The system allows for additional phenotyping of proliferating cells.
- Apoptosis parameters enhance the validity of proliferation indices.
Conclusions:
- Individual in-vitro immune reactivity patterns are distinct and remarkably constant in healthy individuals.
- Immunofunctional test results remain stable for at least 6 months.
- Stable immune response patterns suggest a possible inherent, potentially genetic, characteristic of the individual.
