Multiplex mapping of CD4 T cell epitopes using class II tetramers

Clin Immunol. 2006 Jul;120(1):21-32. doi: 10.1016/j.clim.2006.03.008. Epub 2006 May 4.


With the advent of class II tetramer technology, a tetramer-guided epitope mapping (TGEM) technique was developed for the identification of CD4+ T cell epitopes. This allowed the direct identification of epitopes recognized by the responding T cells, which were restricted to the single MHC allele of interest. However, as each individual carries multiple class II alleles, it would be advantageous to design an approach to identify CD4+ epitopes presented by different class II alleles at the same time. In the present study, a multiplex TGEM approach was developed to identify antigenic epitopes presented by multiple HLA class II alleles simultaneously. In this new approach, CD4+ T cells were stained with multiple sets of MHC class II tetramers-each labeled with a unique fluorescent label. Using this multiplex approach, novel epitopes from influenza antigens hemagglutinin and matrix protein presented by multiple class II alleles were identified in a single experimental setting.

Publication types

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

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • CD4-Positive T-Lymphocytes / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping / methods*
  • Epitopes, T-Lymphocyte / immunology*
  • Flow Cytometry
  • Hemagglutinins / immunology
  • Histocompatibility Antigens Class II / immunology*
  • Humans
  • Influenza A Virus, H1N1 Subtype / immunology
  • Molecular Sequence Data
  • Peptide Fragments / immunology
  • Sensitivity and Specificity
  • Viral Matrix Proteins / immunology


  • Epitopes, T-Lymphocyte
  • Hemagglutinins
  • Histocompatibility Antigens Class II
  • Peptide Fragments
  • Viral Matrix Proteins