Reliable generation and use of MHC class II:gamma2aFc multimers for the identification of antigen-specific CD4(+) T cells

J Immunol Methods. 2002 Dec 20;271(1-2):137-51. doi: 10.1016/s0022-1759(02)00343-5.


MHC tetramers have proven to be powerful reagents for the analysis of MHC class I-restricted T cells. However, generating similarly reliable reagents for MHC class II-restricted T cells has been elusive. Here we evaluated the utility of MHC class II:gamma2aFc multimers, which contain the MHC class II extracellular domains, with or without recombinantly attached peptides, dimerized via a fos-jun leucine zipper and attached to the hinge of murine IgG2a. We have successfully generated 24 multimers in either myeloma or Drosophila melanogaster S2 cells, with an average yield of 7 mg/L. 'Empty' MHC class II:gamma2aFc multimers were effectively used in peptide binding assays. Similar versions that contained recombinantly attached peptides stimulated T cells in an antigen-specific, MHC-restricted manner, and identified antigen-specific nai;ve and effector T cells by flow cytometry. Furthermore, we have successfully used these reagents to stain T cells generated following viral infection. Thus, MHC class II:gamma2aFc multimers are robust and reliable reagents for the analysis of MHC class II-restricted T cells.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • CD4-Positive T-Lymphocytes / immunology*
  • Drosophila melanogaster
  • Epitopes, T-Lymphocyte / immunology*
  • Female
  • Flow Cytometry
  • Histocompatibility Antigens Class II / immunology*
  • Immunoglobulin Fragments / immunology*
  • Influenza A virus / immunology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Molecular Sequence Data
  • Plasmids / immunology
  • Recombinant Proteins / immunology
  • Sendai virus / immunology
  • Specific Pathogen-Free Organisms
  • Tumor Cells, Cultured


  • Epitopes, T-Lymphocyte
  • Histocompatibility Antigens Class II
  • Immunoglobulin Fragments
  • Recombinant Proteins