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International Immunology Advance Access originally published online on June 2, 2006
International Immunology 2006 18(7):1139-1145; doi:10.1093/intimm/dxl048
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© The Japanese Society for Immunology. 2006. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Impact of CD8–MHC class I interaction in detection and sorting efficiencies of antigen-specific T cells using MHC class I/peptide multimers: contribution of pMHC valency

Berangere Neveu1, Klara Echasserieau1, Timothy Hill3, Kristine Kuus-Reichel3, Elisabeth Houssaint1,2, Marc Bonneville1 and Xavier Saulquin1,2

1 Département de Recherche en Cancérologie, INSERM U601, Nantes, France
2 Faculté des Sciences, Université de Nantes, Nantes, France
3 Beckman Coulter, Inc., 7330 Carroll Road, San Diego, CA 92121, USA

Correspondence to: X. Saulquin; E-mail: saulquin{at}nantes.inserm.fr

Recombinant soluble multimeric forms of MHC class I molecules loaded with antigenic peptides (pMHC) have turned out to be particularly useful to detect and isolate specific T cells. These applications both rely on the oligomerization of pMHC monomers in order to compensate for their inherent low affinity for the TCR. In this study, we have evaluated the precise contribution of CD8–pMHC interaction on the specificity and sorting efficiency of pMHC multimers according to their degree of oligomerization. To this end several wild-type versus mutated pMHC complexes (A*0201, B*0701, B*0801, B*3501) carrying point mutations known to reduce (245V mutants) or to abrogate (227,8KA mutants) CD8–pMHC interaction and showing various degrees of valency have been used. We show that irrespective of the HLA allele tested, 245V mutation strongly improves the binding specificity and immunomagnetic sorting efficiency of pMHC multimers. Our results also indicate that the contribution of CD8 to the binding of pMHC multimers to specific CD8+ T cells is inversely correlated to the degree of pMHC oligomerization. Consequently, efficient staining or specific sorting of high-affinity T cells (i.e. CD8 independent) can only be achieved using 227,8KA pMHC complexes with low-order oligomerization.

Keywords: magnetic sorting, pMHC multimers, T cells, tetramers

Transmitting editor: E. Vivier


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