TCR characterization and cytokine profile of MELOE-1 specific CD4 T cell clones.

aCD4 T cell clones were stimulated for 5 hours in the presence of brefeldin A (10 µg/mL) either with the cognate peptide (10 µM) in an autopresentation assay. After 5 hours of stimulation, cells were stained with APC-conjugated anti-CD4 mAb, fixed with 4% paraformaldehyde, labeled with PE-conjugated anti-cytokine mAb and analyzed by flow cytometry. A “high” cytokine production was defined with a threshold of 20% of producing CD4 T cells, in response to the peptide.