A: Schematic of peptides synthesized for testing minimal
epitope. B: A ‘WED’-specific CD4+ T-cell
clone (wb688 clone 1,3) generated from a healthy donor, following ex
vivo peptide-stimulated PBMC expansion, was incubated with an
HLA-DRB1*0401 LCLs and stimulated with DMSO vehicle control, ‘WED’
20-mer (positive control), or internal 11-mer peptides overlapping by 10 amino
acids within the ‘WED’ 20-mer. The absence of responses to some
11-mer peptides provides an addition, internal negative control. Peptides were
tested at 1 μg/mL and IFNγ responses measured by ICS assay. The
percent of viable CD4+IFNγ+ T cells (negative for
CD8/CD14/CD19) in the lymphocyte forward/side scatter region are denoted. These
data were representative of DRB1*0401-restricted WEDLT209–228
-specific CD4+ T-cell clones generated from three individuals as
detailed using a secreted IFNγ readout in Supplemental Figure S5A.