Parallel detection of antigen-specific T cell responses by combinatorial encoding of MHC multimers (original) (raw)

References

  1. Hadrup, S.R. et al. Parallel detection of antigen-specific T cell responses by multidimensional encoding of MHC multimers. Nat. Methods 6, 520–526 (2009).
    Article CAS Google Scholar
  2. Rodenko, B. et al. Generation of peptide-MHC class I complexes through UV-mediated ligand exchange. Nat. Protoc. 1, 1120–1132 (2006).
    Article CAS Google Scholar
  3. Toebes, M. et al. Design and use of conditional MHC class I ligands. Nat. Med. 12, 246–251 (2006).
    Article CAS Google Scholar
  4. Bakker, A.H. et al. 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. USA 105, 3825–3830 (2008).
    Article CAS Google Scholar
  5. Grotenbreg, G.M. et al. Discovery of CD8+ T cell epitopes in Chlamydia trachomatis infection through use of caged class I MHC tetramers. Proc. Natl. Acad. Sci. USA 105, 3831–3836 (2008).
    Article CAS Google Scholar
  6. Frickel, E.M. et al. Parasite stage-specific recognition of endogenous _Toxoplasma gondii_-derived CD8+ T cell epitopes. J. Infect. Dis. 198, 1625–1633 (2008).
    Article CAS Google Scholar
  7. Unger, W.W. et al. Discovery of low-affinity preproinsulin epitopes and detection of autoreactive CD8 T-cells using combinatorial MHC multimers. J. Autoimmun. 37, 151–159 (2011).
    Article CAS Google Scholar
  8. Broen, K. et al. Concurrent detection of circulating minor histocompatibility antigen-specific CD8 T cells in SCT recipients by combinatorial encoding MHC multimers. PLoS ONE 6, e21266 (2011).
    Article CAS Google Scholar
  9. Hombrink, P. et al. High-throughput identification of potential minor histocompatibility antigens by MHC tetramer-based screening: feasibility and limitations. PLoS ONE 6, e22523 (2011).
    Article CAS Google Scholar
  10. Velthuis, J.H. et al. Simultaneous detection of circulating autoreactive CD8+ T-cells specific for different islet cell-associated epitopes using combinatorial MHC multimers. Diabetes 59, 1721–1730 (2010).
    Article CAS Google Scholar
  11. Sick Andersen, R. et al. Dissection of T cell antigen specificity in human melanoma. Cancer Res. published online, doi:10.1158/0008-5472.CAN-11-2614 (6 February 2012).
  12. Kvistborg, P. et al. TIL therapy broadens the tumor-reactive CD8+ T cell compartment in melanoma patients. OncoImmunology 1, 1–10 (2012).
    Article Google Scholar
  13. Chen, D.S. et al. Marked differences in human melanoma antigen-specific T cell responsiveness after vaccination using a functional microarray. PLoS Med. 2, e265 (2005).
    Article Google Scholar
  14. Deviren, G., Gupta, K., Paulaitis, M.E. & Schneck, J.P. Detection of antigen-specific T cells on p/MHC microarrays. J. Mol. Recognit. 20, 32–38 (2007).
    Article CAS Google Scholar
  15. Kwong, G.A. et al. Modular nucleic acid assembled p/MHC microarrays for multiplexed sorting of antigen-specific T cells. J. Am. Chem. Soc. 131, 9695–9703 (2009).
    Article CAS Google Scholar
  16. Bendall, S.C. et al. Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum. Science 332, 687–696 (2011).
    Article CAS Google Scholar
  17. Newell, E.W., Klein, L.O., Yu, W. & Davis, M.M. Simultaneous detection of many T cell specificities using combinatorial tetramer staining. Nat. Methods 6, 497–499 (2009).
    Article CAS Google Scholar
  18. Lissina, A. et al. Protein kinase inhibitors substantially improve the physical detection of T-cells with peptide-MHC tetramers. J. Immunol. Methods 340, 11–24 (2009).
    Article CAS Google Scholar
  19. Bakker, A.H. & Schumacher, T.N. MHC multimer technology: current status and future prospects. Curr. Opin. Immunol. 17, 428–433 (2005).
    Article CAS Google Scholar
  20. Perfetto, S.P. et al. Quality assurance for polychromatic flow cytometry. Nat. Protoc. 1, 1522–1530 (2006).
    Article CAS Google Scholar

Download references

Acknowledgements

We thank T. Seremet (Center for Cancer Immune Therapy, University Hospital Herlev) and M. Toebes (Netherlands Cancer Institute) for excellent technical assistance; and S. Walter (Immatics) and C. Gouttefangeas (Tübingen University) for supporting data on MHC multimer storage. This work was supported by the Danish Cancer Society (grant DP06031), The Danish Council for Strategic Research (grant 09-065152), the Center for Translational Molecular Medicine (grant 04I-301) and Integration of Biosynthesis and Organic Synthesis (grant 053.63.015).

Author information

Author notes

  1. Sine Reker Hadrup
    Present address: Institute for International Health, Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark. Present addresses: Division of Immunology and Pathogenesis, Department of Molecular and Cellular Biology, University of California, Berkeley, California, USA (A.H.B.); Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA (C.J.S.).,

Authors and Affiliations

  1. Department of Hematology, Center for Cancer Immune Therapy, University Hospital Herlev, Herlev, Denmark
    Rikke Sick Andersen, Thomas Mørch Frøsig, Natasja W Pedersen, Rikke Lyngaa, Per thor Straten & Sine Reker Hadrup
  2. Division of Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands
    Pia Kvistborg, Arnold H Bakker, Chengyi Jenny Shu & Ton N Schumacher
  3. Department of Pharmacology and Pharmacotherapy, Pharmaceutical Faculty, University of Copenhagen, Copenhagen, Denmark
    Thomas Mørch Frøsig

Authors

  1. Rikke Sick Andersen
    You can also search for this author inPubMed Google Scholar
  2. Pia Kvistborg
    You can also search for this author inPubMed Google Scholar
  3. Thomas Mørch Frøsig
    You can also search for this author inPubMed Google Scholar
  4. Natasja W Pedersen
    You can also search for this author inPubMed Google Scholar
  5. Rikke Lyngaa
    You can also search for this author inPubMed Google Scholar
  6. Arnold H Bakker
    You can also search for this author inPubMed Google Scholar
  7. Chengyi Jenny Shu
    You can also search for this author inPubMed Google Scholar
  8. Per thor Straten
    You can also search for this author inPubMed Google Scholar
  9. Ton N Schumacher
    You can also search for this author inPubMed Google Scholar
  10. Sine Reker Hadrup
    You can also search for this author inPubMed Google Scholar

Contributions

R.S.A. designed and performed experiments, analyzed data and wrote the paper; P.K. designed and performed experiments, analyzed data and co-wrote the paper; T.M.F. performed experiments and analyzed data; N.W.P. performed experiments and analyzed data; R.L. performed experiments and analyzed data; A.H.B. designed and performed experiments, and analyzed data; C.J.S. designed and performed experiments, and analyzed data; P.t.S. co-wrote the paper; T.N.S. conceived the approach, helped design experiments and co-wrote the paper; S.R.H. designed and performed experiments, analyzed data and wrote the paper.

Corresponding author

Correspondence toSine Reker Hadrup.

Ethics declarations

Competing interests

The technology described in this article is the subject of a patent application. On the basis of the Netherlands Cancer Institute policy on management of intellectual property, S.R.H., A.H.B, C.J.S. and T.N.S would be entitled to a portion of received royalty income.

Supplementary information

Supplementary Figure 1

Experimental setup in 384-well microplates. When multiple MHC multimer panels are generated simultaneously, we advise the use of the outlined setup. Each row on a 384 well plate (plus 3 additional wells on a second plate) holds one full panel of 27 MHC multimers with different color codes. The exchange reaction (step 4) is conducted in 384-well microplates, numbers indicate the different peptides. After the exchange reaction is completed pMHC monomers are transferred to two new 384-well microplates (step 7) for coupling to the different fluorochrome-streptavidin conjugates (step 8). Importantly, a given pMHC complex should always maintain the same position in the plate. Finally, the MHC multimers are mixed in a new plate to obtain the final color codes (step 12), such that each well contains one pMHC complex, conjugated to two different fluorochromes. Before T-cell staining each row is mixed to generate one complete panel of 27 MHC multimers for combinatorial encoding. (PDF 23 kb)

Supplementary Figure 2

Flow cytometry analysis of a T-cell population containing MART-1-reactive T cells that was either stained with MHC (HLA-A0201) multimers made with MART-1 modified peptide (left), or with MART-1 wild type peptide (right). The MART-1 wildtype peptide has a very low affinity for HLA-A0201, whereas the MART-1 modified peptide binds with high affinity. Dot plots are gated on CD8+ T cells and the MART-1-MHC multimer stained T cells are indicated in red. The panel used for this analysis had been stored at 4°C in the dark for six days prior to T-cell staining. Note that staining intensity is comparable for the two peptides. (PDF 11 kb)

Supplementary Table 1

Fluorochrome specifications (PDF 6 kb)

Supplementary Table 2

Typical PMT voltages for an experiment (PDF 5 kb)

Supplementary Table 3

Spectral overlap values from a typical compensation made with the PMT voltages shown in Supplementary Table 2 (PDF 10 kb)

Rights and permissions

About this article

Cite this article

Andersen, R., Kvistborg, P., Frøsig, T. et al. Parallel detection of antigen-specific T cell responses by combinatorial encoding of MHC multimers.Nat Protoc 7, 891–902 (2012). https://doi.org/10.1038/nprot.2012.037

Download citation