Reliable generation and use of MHC class II:γ2aFc multimers for the identification of antigen-specific CD4+ T cells

Paula Y. Arnold, Kate M. Vignali, Timothy B. Miller, Nicole L. La Gruta, Linda S. Cauley, Laura Haynes, P. Scott Adams, Susan L. Swain, David L. Woodland, Dario A.A. Vignali

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23 Citations (Scopus)


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:γ2aFc 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:γ2aFc 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 naïve 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:γ2aFc multimers are robust and reliable reagents for the analysis of MHC class II-restricted T cells.

Original languageEnglish
Pages (from-to)137-151
Number of pages15
JournalJournal of Immunological Methods
Issue number1-2
Publication statusPublished - 20 Dec 2002
Externally publishedYes


  • Flow cytometry
  • Immunoglubulin fusion proteins
  • MHC class II
  • Peptide binding assay
  • T cells
  • Tetramers

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