Recombinant human interleukin il 2

PBMCs were cultured (2×10⁶ cells/well) with zoledronate (5 μM) (Novartis Pharma, Basel, Switzerland) and CMV or GPC3 peptide (10 μM) in AIM-V medium (Gibco) supplemented with 10% human AB serum (Sigma) and recombinant human interleukin (IL)-2 (1,000 IU/ml) (Novartis Pharma) for 14 days. The stimulation procedure was performed at 37°C and 5% CO₂. Scale-up of cells was performed in accordance with their growth.

NK92-cells were used as CAR-effector cells. Daudi and Karpas422 cells (CD19⁺ non-Hodgkin lymphoma cell lines), human primary B cells and Jurkat cells (CD19⁻) served as target cells. All cell lines were authenticated by STR analysis.
NK92-cells were cultured in RPMI1640 (GIBCO) supplemented with 10% FBS (GIBCO), 4 mM L-glutamine (GIBCO), 100 U/ml/100 µg/ml penicillin/streptomycin (Sigma-Aldrich), 10 mM HEPES (Sigma-Aldrich) and 100 U/ml recombinant human Interleukin (IL)-2 (Novartis). Other cell lines were cultured in RPMI1640, containing 2 mM L-glutamine and 50 U/ml/50 µg/ml penicillin/streptomycin supplemented with 20% FBS for Karpas422, 10% FBS for Jurkat or 10% FBS, 1 mM sodium pyruvate (GIBCO) and 10 mM HEPES for Daudi.