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Design and use of conditional MHC class I ligands

Nature Medicine volume 12, pages 246–251 (2006)Cite this article

Abstract

Major histocompatibility complex (MHC) class I molecules associate with a variety of peptide ligands during biosynthesis and present these ligands on the cell surface for recognition by cytotoxic T cells. We have designed conditional MHC ligands that form stable complexes with MHC molecules but degrade on command, by exposure to a defined photostimulus. 'Empty MHC molecules' generated in this manner can be loaded with arrays of peptide ligands to determine MHC binding properties and to monitor antigen-specific T-cell responses in a high-throughput manner. We document the value of this approach by identifying cytotoxic T-cell epitopes within the H5N1 influenza A/Vietnam/1194/04 genome.

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Figure 1: Photocleavage strategy.
Figure 2: UV light–mediated peptide exchange.
Figure 3: T-cell staining with MHC exchange tetramers.
Figure 4: High-throughput screen of H5N1 T-cell epitopes.

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Acknowledgements

We would like to thank V. Cerundolo (Weatherall Institute of Molecular Medicine) for the A2Kb expression construct and F. Lemonnier (Institut Pasteur) for HLA-A2.1-transgenic mice. We would like to thank A. Pfauth and F. van Diepen for flow cytometry assistance, I. Blonk for help in generation of H5N1 DNA vaccines and H. Hilkmann for peptide synthesis. This work was funded by Netherlands Organization for Scientific Research (NWO) Pioneer (to T.S.) and Vidi (to H.O.) grants.

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Author notes

  1. Miriam Coccoris and Adriaan Bins: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Immunology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066 CX, The Netherlands

    Mireille Toebes, Miriam Coccoris, Adriaan Bins, Raquel Gomez, Willeke van de Kasteele, John B A G Haanen & Ton N M Schumacher

  2. Division of Cellular Biochemistry, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066 CX, The Netherlands

    Boris Rodenko & Huib Ovaa

  3. Department of Virology and WHO National Influenza Center, Erasmus Medical Center, P.O. Box 1738, Rotterdam, 3000 DR, The Netherlands

    Nella J Nieuwkoop & Guus F Rimmelzwaan

Authors
  1. Mireille Toebes
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Corresponding authors

Correspondence to Huib Ovaa or Ton N M Schumacher.

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Competing interests

The technology described in this manuscript is the subject of a patent application. Based on Netherlands Cancer Institute policy on management of intellectual property, Mireille Toebes, Huib Ovaa and Ton N. M. Schumacher would be entitled to a portion of received royalty income in case of future licensing.

Supplementary information

Supplementary Fig. 1

Conditional ligands for 2.1 and H-2Db. (PDF 739 kb)

Supplementary Fig. 2

Conditional peptide-MHC complexes are sensitive to UV light. (PDF 621 kb)

Supplementary Fig. 3

Sensitivity and specificity of MHC exchange tetramers. (PDF 925 kb)

Supplementary Fig. 4

Detection of antigen-specific T cells with MHC-immunoglobulin dimers and MHC tetramers. (PDF 3140 kb)

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Toebes, M., Coccoris, M., Bins, A. et al. Design and use of conditional MHC class I ligands. Nat Med 12, 246–251 (2006). https://doi.org/10.1038/nm1360

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