Phage-displayed libraries of peptide/major histocompatibility complexes

Eur J Immunol. 2004 Feb;34(2):598-607. doi: 10.1002/eji.200324721.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacteriophages / immunology*
  • Bacteriophages / metabolism
  • Capsid Proteins
  • DNA-Binding Proteins / immunology
  • DNA-Binding Proteins / metabolism
  • Epitopes
  • H-2 Antigens / immunology*
  • H-2 Antigens / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Peptide Fragments / immunology*
  • Peptide Fragments / metabolism
  • Peptide Library*
  • Receptors, Antigen, T-Cell / immunology
  • Receptors, Antigen, T-Cell / metabolism
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism
  • Viral Fusion Proteins / immunology
  • Viral Fusion Proteins / metabolism
  • beta 2-Microglobulin / immunology
  • beta 2-Microglobulin / metabolism

Substances

  • Capsid Proteins
  • DNA-Binding Proteins
  • Epitopes
  • H-2 Antigens
  • Peptide Fragments
  • Peptide Library
  • Receptors, Antigen, T-Cell
  • Recombinant Fusion Proteins
  • Viral Fusion Proteins
  • beta 2-Microglobulin