HLA-DRB10410-Restricted Recognition of XAGE-1b sub(37-48) Peptide by CD4 T Cells

XAGE-1b belongs to cancer/testis (CT antigens, and has been shown to be expressed frequently in lung cancers and to elicit an antibody response in patients with XAGE-1b-expressing tumors. In this study, we investigated an XAGE-1b peptide recognized by CD4 T cells. CD4 T cells were purified from PBMC of a healthy donor and stimulated with pooled 25-mer peptides overlapped with 15 amino acids spanning the entire XAGE-1b protein. The generation of XAGE-1b-specific CD4 T cells was shown by IFNgamma secretion assay. A CD4 T cell clone OHD1 was obtained by limiting dilution. OHD1 recognized two overlapping peptides, XAGE1-b sub(33-49) and XAGE-1b sub(37-52), by ELISPOT assay. A peptide XAGE-1b sub(38-46) which was included in both XAGE-1b sub(33-49) and XAGE-1b sub(37-52) was predicted to be a DRB1*0410-restricted 9-mer peptide by a computer-based program. We identified the 12-mer peptide XAGE-1b sub(37- 48) as a new XAGE-1b epitope restricted to HLA-DRB1*0410.