Mixed lymphocyte reaction (MLR)

The collected DCs served as stimulator cells, and T lymphocytes served as responder cells in the MLR. T lymphocytes were obtained by passing the isolated human peripheral blood mononuclear cells through a nylon wool column, and then they were co-cultured with the collected DCs in a 5% CO₂ humidified environment at 37°C for 96 h. The ratio of stimulators to responders was 1:10. A cell counting kit-8 assay was conducted to detect T lymphocyte proliferation. The ability of DCs to interact with proliferating T lymphocytes was assessed by measuring the absorbance at 450 nm, which was then normalized to that of the Con A-positive control, producing a ratio.