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

European Journal of Immunology
Fabrice LemaîtreLaurent Ferradini

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 degrees 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 x 10(-6) among peripheral blood mononuclear cells. Such a combined approach may have promising applications in the immunomonitoring of patients under vaccination protocols to tightly follow induced antigen-specific CD4(+) T cells expressing previously identified TCR.

Citations

Apr 18, 2009·Nature Protocols·James J MoonMarc K Jenkins
Mar 3, 2010·PLoS Pathogens·Benoît VingertUNKNOWN ANRS EP36 HIV Controllers Study Group
Nov 23, 2006·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Christian VeldmanMichael Hertl
Jul 10, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Virginia CecconiGiulia Casorati
Jun 22, 2013·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Petra Bacher, Alexander Scheffold
Apr 21, 2012·The Journal of Immunology : Official Journal of the American Association of Immunologists·Marc K Jenkins, James J Moon

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