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. 2009 Nov:Chapter 18:18.16.1-18.16.20.
doi: 10.1002/0471142735.im1816s87.

Generation of peptide MHC class I monomers and multimers through ligand exchange

Mireille Toebes  1 , Boris Rodenko  1 , Huib Ovaa  1 , Ton N M Schumacher  1
Affiliations

Generation of peptide MHC class I monomers and multimers through ligand exchange

Mireille Toebes et al. Curr Protoc Immunol. 2009 Nov.

Abstract

The recognition of defined antigen-MHC complexes by antigen-specific T cells forms the molecular basis of T cell immunity. It has been shown that fluorescently labeled recombinant MHC tetramers can be utilized to detect antigen-specific T cells by flow cytometry. Since this first description, MHC tetramers and other types of MHC multimers have become a core tool to monitor the development of disease- and therapy-induced antigen-specific T cell responses both in humans and in animal model systems. This unit describes a set of protocols that transform classical MHC multimer technology into a high-throughput platform, allowing one to produce large collections of MHC class I molecules charged with different peptides. This technology is based on the development of conditional MHC ligands that can be triggered to self-destruct while in the MHC-bound state.

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References

Literature Cited

References
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    1. Bakker, A.H. and Schumacher, T.N. 2005. MHC multimer technology: Current status and future prospects. Curr. Opin. Immunol. 17:428-433.
    1. Bakker, A.H., Hoppes, R., Linnemann, C., Toebes, M., Rodenko, B., Berkers, C.R., Hadrup, S.R., van Esch, W.J., Heemskerk, M.H., Ovaa, H., and Schumacher, T.N. 2008. Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7. Proc. Natl. Acad. Sci. U.S.A. 105:3825-3830.
    1. Celie, P.H., Toebes, M., Rodenko, B., Ovaa, H., Perrakis, A., and Schumacher, T.N. 2009. UV-induced ligand exchange in MHC class I protein crystals. J. Am. Chem. Soc. 131:12298-12304.
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Key Reference
    1. Toebes et al., 2006. See above.

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