Mcf 7

Human breast carcinoma cell lines MCF-7 and HCC1143, pancreas adenocarcinoma cell line PA-TU-8902, prostate >carcinoma cell line DU145 and lung carcinoma cell line NCI-H460 were obtained from DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany).
In general RPMI 1640 medium, supplemented with 2 mM L-Glutamine and 1% Penicillin-Streptomycin and FCS (either 10% or 1% according to the assay) was used for monocyte/macrophage, NCI-H460, HCC1143 and DU145 cell culture. For MCF-7 cells we used Eagle´s MEM, supplemented with 10% FCS, 2 mM L-Glutamine, 1% Penicillin/Streptomycin and 1 mM Sodium Pyruvate and for PA-TU-8902 cells Dulbecco´s MEM High Glucose supplemented with 1 mM Sodium Pyruvate, 2 mM L-Glutamine, 1% Penicillin/Streptomycin and either 10% or 1% FCS was used (all reagents from Sigma, Buchs, Switzerland).
Cell lines were kept under standard culture conditions (37°C, 5% CO₂ and 95% humidity), tumour cell conditioned media were obtained by 24 h incubation of 90% confluent tumour cells in 10 ml of fresh culture medium in 25 cm² cell flasks. Cell free supernatants were carefully harvested, filtered through 0.5 μm filters and stored in aliquots at −80°C.

All cell culture reagents were purchased from Biowest and Serana. Tumor cell lines ZR-75-1, MCF-7, HSC-4, CaoV-3 and T-47D as well as the human NK cell line KHYG-1 and murine T cell line CTLL-2 were obtained from DSMZ or ATCC and cultured in the respective recommended medium under standard conditions. KHYG-1 and CTLL-2 were cultured in presence of 10 ng/mL IL-2 (PeproTech, Rocky Hill, NJ, USA).
For in vivo studies, the murine tumor cell lines B16.F10 and CT26.wt (ATCC) were stably transfected by electroporation (Amaxa) with a plasmid coding for human MUC1 (40 tandem repeats) and confirmed for TA-MUC1 expression in vitro and in vivo in mice by flow cytometry and immunohistochemistry using GT-00A. Cells were cultured in standard medium +100 nM methotrexate (Hexal, Holzkirchen, Germany) as selection pressure. For the metastasis model, hMUC1-B16.F10 cells were further transduced to stably express firefly luciferase (110 RLU/cell) enabling detection of metastasis. Origin, TA-MUC1 and IL-15R expression status of cell lines are indicated in Table S1.
Peripheral blood mononuclear cells (PBMCs) were prepared from commercially available buffy coats (DRK Berlin) or leukapheresates (Charité Berlin) of healthy donors by Biocoll separation (Biochrom, Cambridge, UK) and density gradient centrifugation. PBMCs from leukapheresate products were stored frozen in liquid nitrogen.

The MYCN-amplified neuroblastoma cell line UKF-NB-3 was established from a stage 4 neuroblastoma patient^{25 (link)}. The vincristine-resistant UKF-NB-3 sub-line UKF-NB-3^rVCR^{10 (link)} (adapted to growth in the presence of vincristine 10 ng/mL) was established by previously described methods^{25 (link)} and derived from the Resistant Cancer Cell Line (RCCL) collection (https://research.kent.ac.uk/industrial-biotechnology-centre/the-resistant-cancer-cell-line-rccl-collection/)³⁷ . The oesophageal cancer cell line COLO680N was obtained from ATCC (Manassas, VA, USA) and the ovarian cancer (COLO-704 and EFO-21) and breast cancer (BT-474 and MCF-7) cell lines from DSMZ (Braunschweig, Germany). All cell lines were propagated in Iscove’s modified Dulbecco’s medium (IMDM) supplemented with 10% FCS, 100 IU/ml penicillin and 100 mg/ml streptomycin at 37 °C. Cells were routinely tested for mycoplasma contamination and authenticated by short tandem repeat profiling.