Combinatorial tetramer staining and mass cytometry analysis facilitate T-cell epitope mapping and characterization

Nat Biotechnol. 2013 Jul;31(7):623-9. doi: 10.1038/nbt.2593. Epub 2013 Jun 9.

Abstract

It is currently not possible to predict which epitopes will be recognized by T cells in different individuals. This is a barrier to the thorough analysis and understanding of T-cell responses after vaccination or infection. Here, by combining mass cytometry with combinatorial peptide-MHC tetramer staining, we have developed a method allowing the rapid and simultaneous identification and characterization of T cells specific for many epitopes. We use this to screen up to 109 different peptide-MHC tetramers in a single human blood sample, while still retaining at least 23 labels to analyze other markers of T-cell phenotype and function. Among 77 candidate rotavirus epitopes, we identified six T-cell epitopes restricted to human leukocyte antigen (HLA)-A*0201 in the blood of healthy individuals. T cells specific for epitopes in the rotavirus VP3 protein displayed a distinct phenotype and were present at high frequencies in intestinal epithelium. This approach should be useful for the comprehensive analysis of T-cell responses to infectious diseases or vaccines.

Publication types

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

MeSH terms

  • Antigens, Viral / immunology
  • Epitope Mapping*
  • Epitopes, T-Lymphocyte / immunology*
  • Flow Cytometry
  • HLA-A Antigens / immunology*
  • Histocompatibility Antigens Class I / immunology
  • Histocompatibility Antigens Class II / immunology
  • Humans
  • Mass Spectrometry
  • Peptides / chemistry
  • Peptides / immunology*
  • Rotavirus / immunology
  • Rotavirus / metabolism
  • T-Lymphocytes

Substances

  • Antigens, Viral
  • Epitopes, T-Lymphocyte
  • HLA-A Antigens
  • Histocompatibility Antigens Class I
  • Histocompatibility Antigens Class II
  • Peptides