title: Phage-displayed libraries of peptide/major histocompatibility complexes.
abstract: Characterizing peptide epitopes targeted by major histocompatibility complex (MHC)-restricted T cells of unknown specificity would have broad implications. In this article we introduce and validate an original phage-displayed library of noncovalent complexes of peptide and MHC (P/MHC). We show that soluble MHC molecules associate with peptides presented by a phage, thereby resulting in the formation of multivalent P/MHC phages. Complex formation is stabilized by the interaction of the soluble partner (MHC) with two components, peptide and beta2-microglobulin, both of which are covalently linked to the phage. As proof of concept, we have used this strategy to express peptide libraries in the context of H-2K(b). Using monoclonal antibody 25D (specific for ovalbumin/H-2K(b)) as a template to screen the library, we were able to select a variant epitope functionally and structurally related to the wild-type peptide. Interaction studies between monoclonal antibody 25D and cells suggest that the variant peptide has been selected on the basis of a decreased dissociation rate between the peptide/H-2K(b) complex and its ligand. A weak agonist of the N15 TCR (vesicular stomatitis virus/H-2K(b)-specific) was also isolated from another P/MHC library. This strategy opens up new perspectives for antigen discovery and the manipulation of T cell responses.
authors.affiliation: INSERM U543, Immunologie A, Hôpital Pitié-Salpêtrière, Paris, France.
authors.forename: Ismail
authors.initials: I
authors.lastname: Dogan
authors.forename: Karim
authors.initials: K
authors.lastname: Dorgham
authors.forename: Hsiu-Ching
authors.initials: HC
authors.lastname: Chang
authors.forename: Christophe
authors.initials: C
authors.lastname: Parizot
authors.forename: Fabrice
authors.initials: F
authors.lastname: Lemaître
authors.forename: Laurent
authors.initials: L
authors.lastname: Ferradini
authors.forename: Ellis L
authors.initials: EL
authors.lastname: Reinherz
authors.forename: Patrice
authors.initials: P
authors.lastname: Debré
authors.forename: Guy
authors.initials: G
authors.lastname: Gorochov
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000276:immunology* / Q000378:metabolism; D036022:Capsid Proteins; D004268:DNA-Binding Proteins / Q000276:immunology / Q000378:metabolism; D000939:Epitopes; D006183:H-2 Antigens / Q000276:immunology* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology* / Q000378:metabolism; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000276:immunology / Q000378:metabolism; D001613:beta 2-Microglobulin / Q000276:immunology / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D000939:Epitopes; D006183:H-2 Antigens; D010446:Peptide Fragments; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins; D001613:beta 2-Microglobulin