Figure 6. First screening of CD4 clones for cytokine secretion upon stimulation with melanocytic proteins. Bacteria coated with complement
were grouped in two pools, added to autologous Epstein-Barr virus-transformed B cells (EBV-B cells), and incubated overnight.
CD4 T cells (5,000/well) were added in monoplicate to EBV-B cells (30,000/well). The amount of tumor necrosis factor (TNF)-α,
interferon (IFN)-γ, interleukin (IL)-5, and IL-17 in the supernatant of overnight co-cultures was estimated with a multiplex
bead-based assay. As the positive control, anti-MAGE-3 T cell clone R12C9 was used.