Bi-specific MHC heterodimers for characterization of cross-reactive T cells

J Biol Chem. 2010 Oct 22;285(43):33144-33153. doi: 10.1074/jbc.M110.141051. Epub 2010 Aug 20.

Abstract

T cell cross-reactivity describes the phenomenon whereby a single T cell can recognize two or more different peptide antigens presented in complex with MHC proteins. Cross-reactive T cells have previously been characterized at the population level by cytokine secretion and MHC tetramer staining assays, but single-cell analysis is difficult or impossible using these methods. In this study, we describe development of a novel peptide-MHC heterodimer specific for cross-reactive T cells. MHC-peptide monomers were independently conjugated to hydrazide or aldehyde-containing cross-linkers using thiol-maleimide coupling at cysteine residues introduced into recombinant MHC heavy chain proteins. Hydrazone formation provided bi-specific MHC heterodimers carrying two different peptides. Using this approach we prepared heterodimers of the murine class I MHC protein H-2K(b) carrying peptides from lymphocytic choriomeningitis virus and vaccinia virus, and used these to identify cross-reactive CD8+ T cells recognizing both lymphocytic choriomeningitis virus and vaccinia virus antigens. A similar strategy could be used to develop reagents to analyze cross-reactive T cell responses in humans.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Viral / immunology*
  • CD8-Positive T-Lymphocytes / immunology*
  • Cell Line
  • Cricetinae
  • Cross Reactions / immunology
  • H-2 Antigens / immunology*
  • Humans
  • Lymphocytic choriomeningitis virus / immunology*
  • Mice
  • Peptides / immunology*
  • Protein Structure, Quaternary
  • Vaccinia virus / immunology*
  • Viral Proteins / immunology*

Substances

  • Antigens, Viral
  • H-2 Antigens
  • H-2Kb protein, mouse
  • Peptides
  • Viral Proteins