Peptide binding to the most frequent HLA-A class I alleles measured by quantitative molecular binding assays.
Quantitative assays to measure the binding of defined synthetic antigenic peptides and purified MHC class I molecules are described for several common human HLA-A alleles (A1, A2.1, A3, A11 and A24). Under appropriate conditions, the binding of radiolabeled peptides to purified MHC class I molecules is very effective, highly specific, and appears to be dependent on the specific sequence motif of the peptide as defined by critical anchor residue positions. Establishment and optimization of the assay reveals that a relatively high fraction of the MHC class I molecules isolated from EBV transformed B cell line sources is capable of binding exogenously added peptide. Scatchard analysis for all alleles yields 5-10% occupancy values. There is a stringent peptide size requirement that is reflected by the direct influence of peptide length on the binding affinity. The peptide-MHC class I interactions demonstrate remarkable similarity to peptide-MHC class II interactions, both in overall affinity and kinetic behavior. The immunological relevance of the peptide-MHC class I binding assay is also demonstrated by measuring the affinity of a panel of previously described HLA restricted peptides for their HLA restriction element. In 91% (10/11) of the cases, the peptides bound with affinities of 50 nM or less, and in the remaining 9% (1/11) of the cases, in the 50 to 500 nM range. Thus, these data provide the first quantitative estimate of what level of HLA-A binding affinity is associated with a diverse panel of immunodominant CTL epitopes in man.
Sette
A
A
Cytel, San Diego, CA 92121.
Sidney
J
J
del Guercio
M F
MF
Southwood
S
S
Ruppert
J
J
Dahlberg
C
C
Grey
H M
HM
Kubo
R T
RT
D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics / Q000276:immunology; D002461:Cell Line, Transformed; D015234:HLA-A Antigens / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*
D015234:HLA-A Antigens; D010455:Peptides