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The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation
Published on: October 13, 2010
Quantitative analysis of lymphocyte differentiation and proliferation in vitro using carboxyfluorescein diacetate
J Hasbold1, A V Gett, J S Rush
1Immune Regulation Group, Medical Foundation, University of Sydney, The Centenary Institute of Cancer Medicine and Cell Biology, Sydney, New South Wales, Australia.
Insights
Mature T and B lymphocytes
Area of Science:
- Immunology and cellular biology, focusing on lymphocyte signaling and behavior.
Background:
- Lymphocyte activation involves receptor-delivered signals that regulate cell fate, crucial for immune regulation.
- Quantitative analysis of in vitro lymphocyte stimulation systems offers insights into lymphocyte behavior.
Purpose of the Study:
- To quantitatively analyze lymphocyte behavior during stimulation.
- To establish a link between lymphocyte differentiation and division number using novel tracking methods.
Main Methods:
- Utilized carboxyfluorescein diacetate succinimidyl ester (CFSE) to track lymphocyte division.
- Incorporated division number as a variable in quantitative analyses of lymphocyte stimulation.
Main Results:
- Established a fundamental link between lymphocyte differentiation and division number.
- Observed independent patterns of change in isotype switching, cytokine expression, and receptor alterations with division number.
Conclusions:
- Lymphocyte differentiation and division number are fundamentally linked.
- Probabilistic modeling is proposed for understanding immune regulation due to stochastic cellular changes and external signal influence.
Abstract:
Mature T and B lymphocytes respond to receptor-delivered signals received during and following activation. These signals regulate the rates of cell death, growth, differentiation and migration that ultimately establish the behaviour patterns collectively referred to as immune regulation. We have been pursuing the philosophy that in vitro systems of lymphocyte stimulation, when analysed quantitatively, help reveal the logical attributes of lymphocyte behaviour. The development of carboxyfluorescein diacetate succinimidyl ester (CFSE) to track division has enabled the variable of division number to be incorporated into these quantitative analyses. Our studies with CFSE have established a fundamental link between differentiation and division number. Isotype switching, expression of T cell cytokines, surface receptor alterations and changes to intracellular signalling components all display independent patterns of change with division number. The stochastic aspects of these changes and the ability of external signals to independently regulate them argue for a probabilistic modelling framework for describing and understanding immune regulation.
