title: 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.
authors.affiliation: Laboratorio de Inmunoquímica I, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Miguel Hidalgo, Mexico.
authors.forename: Claudia E
authors.initials: CE
authors.lastname: Avalos-Martínez
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Rodríguez-Alba
authors.forename: Laura
authors.initials: L
authors.lastname: Berrón-Ruiz
authors.forename: Hector
authors.initials: H
authors.lastname: Romero-Ramírez
authors.forename: Leopoldo
authors.initials: L
authors.lastname: Santos-Argumedo
authors.forename: Luis Antonio
authors.initials: LA
authors.lastname: Jiménez-Zamudio
authors.forename: Maria Lilia
authors.initials: ML
authors.lastname: Domínguez-López
authors.forename: Armando
authors.initials: A
authors.lastname: Vega-López
authors.forename: Ethel
authors.initials: E
authors.lastname: García-Latorre
mesh_terms: D000328:Adult; D001681:Biological Assay; D001973:Bromodeoxyuridine / Q000494:pharmacology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D013936:Thymidine / Q000494:pharmacology; D014316:Tritium
chemical_list: C508017:IL2RA protein, human; D053645:Interleukin-2 Receptor alpha Subunit; D014316:Tritium; D001973:Bromodeoxyuridine; D013936:Thymidine