-Recombinant human TNF-α (R&D Systems, Cat: 210-TA-020), rhIL-2 (Roche, Cat: 11011456001), CFSE (Invitrogen, Cat: C34554), anti-CD3/CD28 beads (Life Technologies, Cat: 111.31D), Anti-Human CD4 PerCP-Cyanine5.5 (eBioscience, Cat: 45-0048), Anti-Human CD25 APC (eBioscience, Cat: 17-0259), Anti-Human CD127 PE (eBioscience, Cat: 12-1278), Human CD4+ T Cell Isolation Kit (Miltenyi Biotec, Cat: 130-096-533), Lymphoprep (Axis-shield, Cat: 1114546)
-Culture medium: RPMI-1640 (Gibco) supplemented with 10% heat-inactivated FBS (Gibco), 2 mM L-glutamine (Gibco), 100 U/ml penicillin and 100 μg/ml streptomycin (Gibco).
-MACS Buffer: 1×PBS supplemented with 0.5% BSA and 2 mM EDTA.
-FACS Buffer: 1×PBS supplemented with 2% FBS.