Summary

Organism		Homo sapiens
Cell		T cell
Cell line		T cell

Characteristics

Experimental design

Previously well characterized primary cultures of lymphocytes purified from healthy blood donors and activated with anti-CD3 and anti-CD28 antibodies for 72 h (Prigent et al., 2014), were used and then exposed to 2 µM B[a]P for the last 48 h.

Additional information

Peripheral blood mononuclear cells were isolated from blood donor buffy coats (written consent for the use of blood samples for the research protocol obtained according to the regulation for blood transfusion of the French blood organization Etablissement Français du Sang, Rennes (France)) by Ficoll (Thermofischer Scientific, Braunschweig, Germany) gradient centrifugation. After separation of monocytes by a 1-h adhesion step, T lymphocytes were purified from nonadherent cells by negative selection using Dynabeads® Untouched™ Human T Cells Kit (Thermofischer Scientific). T lymphocytes were cultured in RPMI medium (Eurobio, Les Ulis, France) supplemented with 20 IU/mL penicillin, 20 μg/mL streptomycin, and 10% decomplemented fetal calf serum (Thermofischer Scientific), and stimulated with Dynabeads® T-Expander beads coated with anti-CD3 and anti-CD28 antibodies (Thermofischer Scientific) before a 48-h treatment with 2 µM B[a]P (Sigma-Aldrich, St. Louis, MO, USA) as previously reported (Liamin et al., 2017). B[a]P was used as stock solutions in dimethylsulfoxide (DMSO). The final concentration of DMSO in culture medium was always < 0.2% v/v and control (CTR) cultures received vehicle containing the same dose of DMSO (CTR) as treated cultures.