Immune Activation by Tumor-Derived DRibbles

Lymphocytes derived from mice spleens were obtained as previously described 10 (link). Briefly, different concentrations (5, 10, 20, 40 μg/ml) of CT-26 cell-derived DRibbles were incubated with lymphocytes for 72 h. To investigate DCs loaded with tumor antigens, 20 μg/ml of CCSC-derived DRibbles (SD) and lysates (SL) and CT-26 cell-derived DRibbles (TD) and lysates (TL) were incubated with the splenic lymphocytes or CD8⁺ T cells (2 × 10⁶ cells/well) for 72 h. Cell viability was assessed using a CCK-8 kit (Beyotime Institute of Biotechnology) according to the manufacturer's instructions. Cells treated with Concanavalin A (5 μg/ml) served as the positive control. Absorbance was measured at 450 nm using a Multiskan Go microplate reader (Thermo Fisher Scientific).