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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.
- Source :
-
International immunology [Int Immunol] 2006 Jul; Vol. 18 (7), pp. 1139-45. Date of Electronic Publication: 2006 Jun 02. - Publication Year :
- 2006
-
Abstract
- 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.
- Subjects :
- Alleles
Amino Acid Substitution
CD8 Antigens chemistry
CD8-Positive T-Lymphocytes chemistry
Histocompatibility Antigens Class I chemistry
Histocompatibility Antigens Class I genetics
Humans
Multiprotein Complexes chemistry
Multiprotein Complexes genetics
Multiprotein Complexes immunology
Peptides chemistry
Peptides genetics
Protein Binding genetics
Receptors, Antigen, T-Cell chemistry
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins immunology
CD8 Antigens immunology
CD8-Positive T-Lymphocytes immunology
Histocompatibility Antigens Class I immunology
Peptides immunology
Receptors, Antigen, T-Cell immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0953-8178
- Volume :
- 18
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- International immunology
- Publication Type :
- Academic Journal
- Accession number :
- 16751231
- Full Text :
- https://doi.org/10.1093/intimm/dxl048