Detection of low-frequency human antigen-specific CD4+ T cells using MHC class II multimer bead sorting and immunoscope analysis

Fabrice Lemaître, Manuelle Viguier, Min Sun Cho, Jean Marie Fourneau, Bernard Maillère, Philippe Kourilsky, Peter M. van Endert, Laurent Ferradini

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

MHC class II tetramers are attractive tools to study antigen-specific CD4+ T cell responses in various clinical situations in humans. HLA-DRA1*0101/DRB1*0401 MHC class II heterodimers were produced as empty molecules using the Drosophila melanogaster expression system. Peptide binding experiments revealed that these molecules could be loaded efficiently with appropriate MHC class II tumor epitopes. Interestingly, MHC class II tetramer staining was influenced by modifications in membrane lipid rafts, and could in itself induce activation changes of stained CD4+ T cells at 37°C. In order to increase the threshold of detection of poorly represented peripheral antigen-specific CD4+ T cells, we combined cell sorting using MHC class II multimer beads together with TCR analysis using the immunoscope technology. This strategy greatly increased the sensitivity of detection of specific CD4+ T cells to frequencies as low as 4 × 10-6 among peripheral blood mononuclear cells. Such a combined approach may have promising applications in the immunomonitoring of patents under vaccination protocols to tightly follow induced antigen-specific CD4+ T cells expressing previously identified TCR.

Original languageEnglish
Pages (from-to)2841-2949
Number of pages109
JournalEuropean Journal of Immunology
Volume34
Issue number10
DOIs
StatePublished - Oct 2004

Keywords

  • HLA-DR4
  • Immunoscope analysis
  • Influenza hemagglutinin-A
  • MHC class II tetramer

Fingerprint

Dive into the research topics of 'Detection of low-frequency human antigen-specific CD4+ T cells using MHC class II multimer bead sorting and immunoscope analysis'. Together they form a unique fingerprint.

Cite this