PBMCs culture
(1x106/ml) cells of PBMCs were cultured, using complete RPMI 1640 supplemented with 2% human serum FPS, 2 m M L-glutamine, 50 units/ml penicillin, and 50 ng/ml streptomycin in the presence of 5µg/ml concanavalin A (Con A) and 50ng/ml interleukin-2 (IL-2). Then cultured cells were incubated at CO2incubator at 37°C in atmospheric pressure of 5% CO2 for 72 hrs. Brefeldin A was added to the cultured cells 3 hours before harvesting [14].