T lymphocytes from healthy donors were isolated by Ficoll method (GE Healthcare, Little Chalfont, UK) followed by positive selection using magnetic beads coated with anti-CD8 antibody (Miltenyi Biotec, Bergisch Gladbach, Germany). Cytotoxic CD8+ T lymphocytes were stimulated with 500 ng/mL ionomycin (Cayman, Ann Arbor, MI, USA; #1932) and 20 ng/mL phorbol 12-myristate-13-acetate (PMA, Adipogen, San Diego, CA, USA; #AG-CN2-0010) for 24 h. Secretions of mouse and human IFN-γ (interferon gamma) were measured by the respective ELISA kits according to the protocols from the manufacturer (eBioscience, San Diego, CA, USA; #88-7314-88 and #88-7316-88).
B16-OVA tumor cells were exposed to cisplatin or oxaliplatin (TargetMol, Boston, MA, USA) for 24 h and then co-cultured with immune cells (B3Z and bone marrow-derived dendritic cells) as previously described [28] (link). Platins were not removed from the culture media in order to expose immune cells to the drugs. Interleukin-2 (IL-2) levels in the culture supernatants were measured by ELISA (eBioscience, San Diego, CA, USA; #88-7024-88).
Mouse CXCL10 (C-X-C Motif Chemokine Ligand 10) protein levels in culture medium were quantified by ELISA according to the protocol provided by the manufacturer (R&D systems, #DY466-05).
B16-OVA tumor cells were exposed to cisplatin or oxaliplatin (TargetMol, Boston, MA, USA) for 24 h and then co-cultured with immune cells (B3Z and bone marrow-derived dendritic cells) as previously described [28] (link). Platins were not removed from the culture media in order to expose immune cells to the drugs. Interleukin-2 (IL-2) levels in the culture supernatants were measured by ELISA (eBioscience, San Diego, CA, USA; #88-7024-88).
Mouse CXCL10 (C-X-C Motif Chemokine Ligand 10) protein levels in culture medium were quantified by ELISA according to the protocol provided by the manufacturer (R&D systems, #DY466-05).