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Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs
Published on: December 27, 2016
A new method for rapid detection of T lymphocyte decision to proliferate after encountering activating surfaces
Elodie Cretel1, Dominique Touchard, Pierre Bongrand
1INSERM UMR 600, Lab Adhésion et Inflammation, Parc Scientifique de Luminy, Case 937, 13288 Marseille Cedex 09, France.
Insights
Measuring T lymphocyte activation is crucial. We found that the contact area between T lymphocytes and surfaces strongly correlates with T cell proliferation, offering a faster assessment method.
Area of Science:
- Immunology
- Cell Biology
- Biophysics
Background:
- T lymphocyte activation is vital for adaptive immunity.
- Detecting rare foreign peptides on antigen-presenting cells is a significant challenge for T cells.
- Current methods for monitoring T cell activation, like proliferation or cytokine synthesis assays, are time-consuming.
Purpose of the Study:
- To identify a rapid and accurate method for assessing T lymphocyte activation.
- To correlate early cellular events with established measures of T cell response.
- To explore potential clinical applications for monitoring lymphocyte reactivity.
Main Methods:
- Quantifying the contact area formed between T lymphocytes and activating surfaces.
- Comparing contact area measurements with standard carboxyfluorescein succinimidyl ester (CFSE) proliferation assays.
- Evaluating the correlation between contact area and Erk activation.
Main Results:
- The contact area between T lymphocytes and activating surfaces showed a strong correlation with T cell proliferative response.
- This correlation was superior to that observed with Erk activation, previously considered a digital reporter.
- The described method provides a simple and accurate way to assess lymphocyte-to-surface contact.
Conclusions:
- Lymphocyte-to-surface contact area is a sensitive and reliable indicator of T cell activation.
- This method offers a faster alternative to traditional assays for monitoring T cell responses.
- The findings suggest potential clinical utility for assessing lymphocyte reactivity and studying early activation events.
Abstract:
A critical step of the adaptive response is the detection of foreign peptides on antigen presenting cells by T lymphocytes. It is a major challenge for a T lymphocyte to detect the presence of a few tens of cognate ligands or less on the membrane of a cell exposing millions of protein molecules. Detection is followed by the cell decision to undergo full or partial activation or even to start an inhibitory program. While the measurement of cell proliferation or cytokine synthesis is accepted as a reliable means of monitoring T lymphocyte activation, this requires hours or days to complete, which is a significant drawback to relate decision to particular signaling events or to assess lymphocyte reactivity in patients. Here we show that the contact area formed between T lymphocytes and potentially activating surfaces is exquisitely correlated to the proliferative response measured with the standard CFSE technique. Correlation is even better than the Erk activation that was reported as a digital reporter of cell activation. The simple and accurate method of assessing lymphocyte-to-surface contact extension that we describe might be very useful both to monitor lymphocyte reactivity for clinical purposes and to identify early steps of lymphocyte activation.
