Whole blood (20 ml) was diluted with 10 ml of phosphate-buffered saline (PBS) + 500 μl of 100 mM EDTA and layered on 20 ml of Percoll. Interface peripheral blood mononuclear cells (PBMCs) (20 min, 300g, room temperature) were washed in PBS and resuspended in 25 ml of T cell medium (X-VIVO 15, Lonza BioWhittaker, Maryland, USA) supplemented with penicillin (100 IU/ml)/streptomycin (100 μg/ml) (Sigma-Aldrich, Missouri, USA) and cultured overnight before use.

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