TY - JOUR
T1 - Detection of low-frequency human antigen-specific CD4+ T cells using MHC class II multimer bead sorting and immunoscope analysis
AU - Lemaître, Fabrice
AU - Viguier, Manuelle
AU - Cho, Min Sun
AU - Fourneau, Jean Marie
AU - Maillère, Bernard
AU - Kourilsky, Philippe
AU - van Endert, Peter M.
AU - Ferradini, Laurent
PY - 2004/10
Y1 - 2004/10
N2 - 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.
AB - 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.
KW - HLA-DR4
KW - Immunoscope analysis
KW - Influenza hemagglutinin-A
KW - MHC class II tetramer
UR - http://www.scopus.com/inward/record.url?scp=7244248655&partnerID=8YFLogxK
U2 - 10.1002/eji.200425281
DO - 10.1002/eji.200425281
M3 - Article
C2 - 15368300
AN - SCOPUS:7244248655
SN - 0014-2980
VL - 34
SP - 2841
EP - 2949
JO - European Journal of Immunology
JF - European Journal of Immunology
IS - 10
ER -