IFNγ ELISpot assays
STERILE
IFNγ ELISpot assays were performed with fresh PBMCs and a human IFNγ ELISpot Kit (Mabtech) following the manufacturer’s instructions. Tests were performed in duplicate. The precoated ELISpot plates were washed with sterile PBS and blocked with RPMI 1640 medium containing 10% fetal bovine serum and 1× penicillin–streptomycin solution (Gibco) for at least 30 min at room temperature. Fresh PBMCs were added at 2 × 10