Identification of an HLA-A11-restricted epitope from the tandem repeat domain of the epithelial tumor antigen mucin.
Epithelial cell mucin encoded by the MUC-1 gene is overexpressed and aberrantly glycosylated on pancreatic, breast, and ovarian cancers as well as on multiple myelomas. It is recognized by patients' Ab and by T cells derived from tumor-draining lymph nodes. The T cell recognition is not MHC restricted and is specific for an epitope previously localized to the immunodominant tandem repeat region of the native mucin molecule. In search of possible MHC-restricted epitopes in the same immunodominant region, we synthesized a panel of overlapping, nine-amino acid long peptides spanning the MUC-1 tandem repeat and first examined their binding to specific human MHC class I molecules using two independent flow cytometry-based assay systems. This approach identified one peptide, p9-17 (STAPPAHGV), that bound to HLA-A1, -A2.1, -A3, and -A11. Measurements of the affinity of binding to each of these alleles, using a quantitative molecular binding assay, indicated that only the relative binding affinity to HLA-A11 was close to immunogenic values. We tested the immunogenicity of p9-17 in vitro. We detected a secondary T cell response specific for p9-17 in lymph nodes from an HLA-A11 breast cancer patient. Moreover, CTL specific for p9-17 peptide could be generated from PBL in several healthy HLA-A11 donors by primary in vitro stimulation.
Doménech
N
N
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, PA 15261, USA.
Henderson
R A
RA
Finn
O J
OJ
D000595:Amino Acid Sequence; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D004848:Epithelium / Q000276:immunology / Q000473:pathology; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D015234:HLA-A Antigens / Q000737:chemistry / Q000276:immunology*; D059870:HLA-A11 Antigen; D006801:Humans; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry / Q000276:immunology*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D012091:Repetitive Sequences, Nucleic Acid; D017421:Sequence Analysis; D014407:Tumor Cells, Cultured
D000939:Epitopes; D015234:HLA-A Antigens; D059870:HLA-A11 Antigen; D009077:Mucins; D010455:Peptides