Cell Lines, Pleural Effusions, and Media

Three NSCLC cell lines (A549, PC9, H1650), the leukemia cell line K562 and 293 T cell line were purchased from the American Type Culture Collection (ATCC, Manassas, VA). Thirteen samples of pleural effusions were obtained from patients diagnosed with lung adenocarcinoma in the First Affiliated Hospital of Zhejiang University. Peripheral blood mononuclear cells (PBMCs) derived from human donors were collected by Ficoll–Hypaque density-gradient centrifugation provided by the Zhejiang Blood Center. All cell lines were grown in RPMI 1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum(FBS) and100 μg/ml penicillin. Medium with recombinant human interleukin-2 (IL-2) of 300 U/ml was used for the expansion of T cells.