Isolation and culture of peripheral T cells

Peripheral blood mononuclear cells (PBMCs) from old cancer patients (more than 60 years old) and young donors (25–30 years old) at Beijing Cancer Hospital (Beijing, China) were obtained by Ficoll-Paque (GE Healthcare, UK) gradient centrifugation, and then peripheral T cells (PTCs) were isolated using a Dynabeads⑥ UntouchedTM Human T Cells Kit (Thermo Fisher Scienti?c, USA) for the following experiments. After ?nishing the test of viability, PBMCs were cultured in T-cell culture medium, which consisted of X-VIVO 15 serum-free medium (Lonza, USA) appended with GlutaMAX (Thermo Fisher, USA), IL2 (50 U mL- 1; Peprotech, USA), IL-7 (10 ng mL- 1; Peprotech, USA), IL-15 (10 ng mL- 1, Peprotech, USA), OKT3 antibody (50 ng mL- 1; ACRO, USA)