Human PBMC proliferation assays

Human peripheral blood mononuclear cells (PBMCs) were obtained from whole blood of one healthy adult volunteer. PBMCs were seeded in 96-well plates at 1 × 10⁵ cells per well in a final volume of 100μL per well, using RPMI + Glutamine media with 5% FBS. Serial dilutions of monoclonal antibodies were then added to the wells containing cells, followed immediately by the addition of SEK or control media. Cells were incubated in 10% CO₂ for 96 hours at 37°C. All treatments were performed in triplicate. Cell proliferation was measured using the Via-Light HS cell proliferation kit (Lonza) according to the manufacturer's instructions.