Peripheral blood mononuclear cells isolation
Human peripheral blood (20 ml) samples were collected from patients and healthy donors in vacutainer tubes (BD Vacutainer tubes with EDTA K2; BD Vacutainer RST tube) through venepuncture; complete blood and serum were obtained. Peripheral blood mononuclear cells (PBMC) were isolated from peripheral venous blood by density gradient centrifugation with Ficoll-Hypaque (Sigma Aldrich). Cellular viability was evaluated by staining with trypan blue. PBMC were used for staining and monocyte isolation.