Proliferation assay
PROLIFERATION
PBMCs were stained with 2.5 μM cell trace violet (CTV, Thermo Fisher) according to user’s manual. T cells were stimulated with 5 μg/mL phytohemagglutinin (PHA) and B cells with a monoclonal CD40 antibody (5 μg/mL; clone: G28.5, BioXCell) and CpG class B (2.5 μM; ODN Invivogen). After 5 days of stimulation, PBMCs were stained using CD4-PE (clone: OKT4), CD8-APC (clone: HIT8a), CD19-PE (clone: HIB19, all Biolegend, San Diego, CA, USA), and fixable viability dye eFluor780 (Thermo Fisher) and proliferation was quantified by flow cytometry, using the MACSQuant 16 analyzer.