Competition-based cellular peptide binding assays for 13 prevalent HLA class I alleles using fluorescein-labeled synthetic peptides.
We report the development, validation, and application of competition-based peptide binding assays for 13 prevalent human leukocyte antigen (HLA) class I alleles. The assays are based on peptide binding to HLA molecules on living cells carrying the particular allele. Competition for binding between the test peptide of interest and a fluorescein-labeled HLA class I binding peptide is used as read out. The use of cell membrane-bound HLA class I molecules circumvents the need for laborious biochemical purification of these molecules in soluble form. Previously, we have applied this principle for HLA-A2 and HLA-A3. We now describe the assays for HLA-A1, HLA-A11, HLA-A24, HLA-A68, HLA-B7, HLA-B8, HLA-B14, HLA-B35, HLA-B60, HLA-B61, and HLA-B62. Together with HLA-A2 and HLA-A3, these alleles cover more than 95% of the Caucasian population. Several allele-specific parameters were determined for each assay. Using these assays, we identified novel HLA class I high-affinity binding peptides from HIVpol, p53, PRAME, and minor histocompatibility antigen HA-1. Thus these convenient and accurate peptide-binding assays will be useful for the identification of putative cytotoxic T lymphocyte epitopes presented on a diverse array of HLA class I molecules.
Kessler
Jan H
JH
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, The Netherlands.
Mommaas
Bregje
B
Mutis
Tuna
T
Huijbers
Ivo
I
Vissers
Debby
D
Benckhuijsen
Willemien E
WE
Schreuder
Geziena M Th
GM
Offringa
Rienk
R
Goulmy
Els
E
Melief
Cornelis J M
CJ
van der Burg
Sjoerd H
SH
Drijfhout
Jan W
JW
D000483:Alleles; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000276:immunology / Q000378:metabolism; D001667:Binding, Competitive; D002461:Cell Line, Transformed; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D015684:Gene Products, pol / Q000276:immunology / Q000378:metabolism; D005805:Genes, MHC Class I; D015488:HIV Antigens / Q000276:immunology / Q000378:metabolism; D015234:HLA-A Antigens / Q000276:immunology / Q000378:metabolism; D015235:HLA-B Antigens / Q000276:immunology / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D020128:Inhibitory Concentration 50; D015778:Minor Histocompatibility Antigens / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D016159:Tumor Suppressor Protein p53 / Q000276:immunology / Q000378:metabolism
D000951:Antigens, Neoplasm; D018984:Epitopes, T-Lymphocyte; D015684:Gene Products, pol; C110615:HA-1 antigen; D015488:HIV Antigens; D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015395:Histocompatibility Antigens Class I; D015778:Minor Histocompatibility Antigens; D009842:Oligopeptides; C404944:PRAME protein, human; D010446:Peptide Fragments; D016159:Tumor Suppressor Protein p53