Phage display of peptide/major histocompatibility complex.
To date, there is no direct way to determine the antigenic specificity of T-cells. While B-cell epitopes can be selected from phage-displayed libraries of peptides, the corresponding molecular tool for identifying T-cell epitopes does not yet exist. The natural ligands of the T-cell antigen-receptor (TCR) are essentially antigenic peptides (P) associated with the products of the major histocompatibility complex (MHC). Here, we report phages displaying P-MHC complexes. Single-chain P-MHC class I molecules, produced in E. coli periplasm, stimulate T-cells in a peptide-specific fashion. The same P-MHC, fused at the tip of filamentous phage, directed their binding to a recombinant TCR restricted to the displayed MHC haplotype (H-2K(d)). Importantly, the binding of P-K(d)-fd to a K(d)-restricted TCR, and also to K(d)-restricted T-cell hybridomas, was modulated by the displayed peptide. Therefore, we suggest phage display of P-MHC as a direct molecular tool for probing T-cell specificity, and for selecting TCR ligands from genetic libraries encoding randomized or natural peptides.
Le Doussal
J
J
Laboratoire d'Immunologie Cellulaire et Tissulaire, UMR CNRS 7627, Hôpital Pitié-Salpetrière, 75013, Paris, France.
Piqueras
B
B
Dogan
I
I
Debré
P
P
Gorochov
G
G
D017951:Antigen Presentation; D036022:Capsid Proteins; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D006183:H-2 Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D009842:Oligopeptides / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D019151:Peptide Library; D019897:Periplasm; D011485:Protein Binding; D011487:Protein Conformation; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D012995:Solubility; D014760:Viral Fusion Proteins / Q000096:biosynthesis
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D000939:Epitopes; D006183:H-2 Antigens; C024755:H-2K(K) antigen; D009842:Oligopeptides; D010446:Peptide Fragments; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins