title: Synthetic peptide libraries in the determination of T cell epitopes and peptide binding specificity of class I molecules.
abstract: Major histocompatibility complex (MHC) class I molecules combine with short peptides of defined length and sequence. Here we describe an approach that may be used in the analysis of peptide preference of different allelic MHC class I molecules, and in the determination of T cell epitopes. We produced synthetic "peptide libraries" of limited complexity by standard peptide chemistry. Using these peptide mixtures we show that H-2 Kb molecules can accommodate both 8- and 9-residue peptides, whereas Db molecules are unable to combine with peptides shorter than 9 amino acids present in these libraries. When these peptide mixtures are used to provide "fingerprints" of Db molecules and mutants thereof, both loss and gain of the ability to combine with certain peptides is observed. For the Kbm1 mutant a strong influence of amino acid substitutions in class I molecules on the peptides selected is observed. In these synthetic peptide mixtures, the presence of a specific T cell epitope, known to be represented once, can be detected. This approach may be extended to the identification of new T cell epitopes from larger peptide libraries.
authors.affiliation: Department of Cellular Biochemistry, The Netherlands Cancer Institute, Amsterdam.
authors.forename: T N
authors.initials: TN
authors.lastname: Schumacher
authors.forename: G M
authors.initials: GM
authors.lastname: Van Bleek
authors.forename: M T
authors.initials: MT
authors.lastname: Heemels
authors.forename: K
authors.initials: K
authors.lastname: Deres
authors.forename: K W
authors.initials: KW
authors.lastname: Li
authors.forename: M
authors.initials: M
authors.lastname: Imarai
authors.forename: L N
authors.initials: LN
authors.lastname: Vernie
authors.forename: S G
authors.initials: SG
authors.lastname: Nathenson
authors.forename: H L
authors.initials: HL
authors.lastname: Ploegh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D002478:Cells, Cultured; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000276:immunology*; D006183:H-2 Antigens / Q000276:immunology* / Q000302:isolation & purification; D007165:Immunosuppression Therapy; D051379:Mice; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D011110:Polymorphism, Genetic; D013601:T-Lymphocytes / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014758:Viral Core Proteins / Q000276:immunology
chemical_list: D000939:Epitopes; D006183:H-2 Antigens; C024755:H-2K(K) antigen; C032876:H-2Kb protein, mouse; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D014758:Viral Core Proteins