Phorbol 12 myristate 13 acetate pma ionomycin calcium salt

Histopaque-1077, Dulbecco’s phosphate buffered saline (DPBS), Roswell Park Memorial Institute medium (RPMI 1640), Fetal calf serum (FCS), Dimethylsulfoxide (DMSO), Penicillin/Streptomycin, 2-mercaptoethanol and Trypan blue 0.4% were all purchased from ThermoFisher (France). Phorbol 12-myristate 13-acetate (PMA) ionomycin calcium salt, Phytohemagglutinin-L (PHA) and Concanavalin A (ConA) were purchased from Sigma-Aldrich (France). Toxoplasma serological test (TOXOPLASMA ICT IgG-IgM) was purchased from LD Bio Diagnostic (France). ELISPOT plates were purchased from Mabtech (Sweden). MicroBCA kit was purchased from Pierce (France). Vero cells were obtained from ATCC (CCL-81), as well as T.gondii tachyzoites (RH strain, ATCC 50174), and stored in liquid nitrogen until use.

Anti-CD25-PE, anti-CD4-FITC, and anti-CD4-APC antibodies were obtained from BD Biosciences—Pharmingen (San Jose, CA, USA); anti-IL-17A-FITC, anti-IFN-γ-PerCP, and anti-Foxp3-APC (FJK-16s) staining kits were from eBiosciences (San Diego, CA, USA); anti-CD80-FITC, anti-CD86-APC and anti-HLA-DR-PE antibodies were obtained from BD Biosciences—Pharmingen. Phorbol 12 myristate 13-acetate (PMA), Ionomycin calcium salt and Brefeldin A (BFA) were from Sigma-Aldrich (Milano, Italy). Purified lipopolysaccharide (LPS) was purchased from Invivogen (San Giuliano Milanese, Milano, Italy). RPMI 1640, antibiotics (penicillin and streptomycin), L-glutamine, heat-inactivated fetal bovine serum were purchased from Celbio (Pero, Italy) and used for cultures with dendritic cells.