Identification of peptide epitopes of MAGE-1, -2, -3 that demonstrate HLA-A3-specific binding.
The MAGE gene family of tumour antigens are expressed in a wide variety of human cancers. We have identified 43 nonamer peptide sequences, from MAGE-1, -2 and -3 proteins that contain binding motifs for HLA-A3 MHC class I molecules. The T2 cell line, transfected with the cDNA for the HLA-A3 gene, was used in a MHC class I stabilisation assay performed at 37 degrees C and 26 degrees C. At 37 degrees C, 2 peptides were identified that stabilised HLA-A3 with high affinity (fluorescence ratio, FR > 1.5), 4 peptides with low affinity (FR 1.11-1.49) and 31 peptides that did not stabilise this HLA haplotype (FR < 1.1). At 26 degrees C, 12 peptides were identified that stabilised HLA-A3 with high affinity, 8 peptides with low affinity and 17 peptides that did not stabilise this HLA haplotype. Two peptides stabilised HLA-A3 at both temperatures. Small changes in one to three amino acids at positions distinct from the anchor residues altered peptide affinity. Data were compared to a similar study in which a peptide competition assay was used to investigate MAGE-1 peptide binding to several HLA haplotypes. This study demonstrates that anchor residues do not accurately predict peptide binding to specific HLA haplotypes, changes in one to three amino acids at positions distinct from anchor residues influence peptide binding and alternative methods of determining peptide binding yield different results. We are currently investigating the ability of these peptides to induce antitumour cytotoxic T lymphocyte activity as they may be of potential therapeutic value.
McIntyre
C A
CA
Institute for Cancer Studies, University of Sheffield Medical School, UK.
Rees
R C
RC
Platts
K E
KE
Cooke
C J
CJ
Smith
M O
MO
Mulcahy
K A
KA
Murray
A K
AK
D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D000939:Epitopes / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D015791:HLA-A3 Antigen / Q000276:immunology / Q000378:metabolism*; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D006801:Humans; D058950:Melanoma-Specific Antigens; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D010455:Peptides / Q000032:analysis / Q000276:immunology / Q000378:metabolism
D000951:Antigens, Neoplasm; D000939:Epitopes; D015791:HLA-A3 Antigen; D015395:Histocompatibility Antigens Class I; C072011:MAGEA1 protein, human; C072013:MAGEA3 protein, human; C072012:MAGEB2 protein, human; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D010455:Peptides