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. Author manuscript; available in PMC: 2020 Apr 1.
Published in final edited form as: Cancer Immunol Res. 2019 Aug 12;7(10):1727–1739. doi: 10.1158/2326-6066.CIR-19-0103

Figure 3: The ‘WED’ minimal epitope presented by HLA-DRB1*0401 encompasses the Rb-binding motif.

Figure 3:

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.