Fig. 1.

Experimental workflow of the in vivo (plasma) and in vitro (PBMC) assays. Plasma and PBMC from subjects were obtained by peripheral blood centrifugation on Ficoll–Hypaque density gradients. The plasmas were submitted to cytokine analysis through Luminex. The PBMC cultures were stimulated for 3 days or 5 days with anti-CD3/anti-CD28 (α-CD3/α-CD28) beads or MBP/hrIL-2, respectively. In some wells, the 5-HT was added. The content of cytokines in the supernatants from those PBMC cultures was determined by Luminex. The T cells phenotypes were determined by cytometry after activating PBMC cultures with PMA/Ionomycin for 3 h