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Efficient detection and immunomagnetic sorting of specific T cells using multimers of MHC class I and peptide with reduced CD8 binding

Nature Medicine volume 6, pages 707–710 (2000)Cite this article

An Erratum to this article was published on 01 January 2001

Tetramers of major histocompatibility complex (MHC) and peptide represent a large technological advance in immunology by allowing direct analysis of antigen-specific T cells without in vitro manipulation1,2,3,4. However, these tetramers show a concentration-dependent background binding to nonspecific T cells by interacting with the co-receptor CD8. We therefore produced tetramers of human leukocyte antigen (HLA)-A0201 and peptides, with a mutation (of alanine to valine at position 245, called V245 here) known to alter the interaction with CD8 (refs. 5 and 6). These mutated tetramers showed a greatly diminished nonspecific binding but retained specific binding, thus allowing accurate discrimination and very efficient immunomagnetic sorting of rare, specific T cells (less than 1%), without requiring the use of monoclonal antibody against CD8. Therefore, alterations of CD8 binding by mutation of the MHC greatly improved the specificity of MHC–peptide multimers, thus providing efficient tools to sort specific human T cells for immunotherapy.

Staining of T-cell clones with tetramers

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Figure 1: Comparative labeling of T-cell clones with native and mutated HLA-A0201–peptide tetramers.
Figure 2: Labeling of two specific T-cell clones with distinct CD8 dependencies with the mutated A2–GLC tetramer.
Figure 3: Detection of specific T cells within a polyclonal population by single or double staining with tetramers.
Figure 4: Characterization of cells sorted with native or mutated tetramers.

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Acknowledgements

We thank A. McMichael (MRC Human Immunology Unit, Oxford) for plasmids pHN1/β2m and pHN1/HLA-A2. M.B. is supported by a fellowship from the Ligue Nationale Contre le Cancer.

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Authors and Affiliations

  1. 9 quai Moncousu, Nantes, INSERM U463, France

    Marie Bodinier, Marie-Alix Peyrat, François Davodeau, Marc Bonneville & François Lang

  2. Immunotech SA, Beckman Coulter Company, Marseilles, France

    Claire Tournay & François Romagne

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Correspondence to François Lang.

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Bodinier, M., Peyrat, MA., Tournay, C. et al. Efficient detection and immunomagnetic sorting of specific T cells using multimers of MHC class I and peptide with reduced CD8 binding. Nat Med 6, 707–710 (2000). https://doi.org/10.1038/76292

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