Measurement of Suppressor Activity of T CD4+CD25+ T Reg Cells Using Bromodeoxyuridine Incorporation Assay

Abstract

The suppressor effect of T regulatory lymphocytes in co-cultures with T effector cells obtained by magnetic columns from healthy donors and activated by CD3/CD28 was measured by a proliferation assay using BrdU incorporation and an ELISA test. Tritiated thymidine incorporation was used as a reference since it is the gold standard for proliferation assays. Both methods were used simultaneously in the same samples in order to compare them. Correlation between them was statistically significant (p < 0.001). The purification using magnetic columns was very efficient since CD4+CD25+ cells were also FOXP3+ therefore; they were identified as suppressor T cells. The use of BrdU incorporation in suppression assays is an excellent method that avoids the use of radioactive contaminating materials.

• Regulatory T cells
• Effector T cells
• Thymidine incorporation
• BrdU incorporation
• Suppression by T reg cell

ACKNOWLEDGEMENTS

C.E. Ávalos-Martínez is a DSc. student who received a doctoral scholarship from CONACyT, México. L. Jiménez-Zamudio, ML Domínguez-López, A.Vega-López and E. García-Latorre are fellows of COFAA and EDI-IPN. The work was done in part with grants from SIP-IPN and CONACyT, México.

Declaration of Interests

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.