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Reversible MHC multimer staining for functional isolation of T-cell populations and effective adoptive transfer

Nature Medicine volume 8, pages 631–637 (2002)Cite this article

Abstract

Recently developed major histocompatibility complex (MHC) multimer technologies allow visualization and isolation of antigen-specific T cells. However, functional analysis and in vivo transfer of MHC multimer-stained cells is hampered by the persistence of T-cell receptor (TCR)–MHC interactions and subsequently induced signaling events. As MHC monomers do not stably bind to TCRs, we postulated that targeted disassembly of multimers into MHC monomers would result in dissociation of surface-bound TCR ligands. We generated a new type of MHC multimers, which can be monomerized in the presence of a competitor, resulting in rapid loss of the staining reagent. Following dissociation, the T cells are phenotypically and functionally indistinguishable from untreated cells. This 'reversible' T-cell staining procedure, which maintains the specificity and sensitivity of MHC multimer staining while preserving the functional status of T lymphocytes, may be of broad benefit for ex vivo investigation of T-cell functions and clinical applications.

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Figure 1: Interference of MHC multimer staining with T-cell function in vitro.
Figure 2: Interference of MHC multimer staining with T-cell function in vivo.
Figure 3: Development of a reversible MHC multimer staining technology.
Figure 4: Improvement of MHC-streptagII-StrepTactin multimer reagents.
Figure 5: Reversible MHC multimer staining and T-cell function.

References

  1. Miyahira, Y. et al. Quantification of antigen specific CD8+ T cells using an ELISPOT assay. J. Immunol. Meth. 181, 45–54 (1995).

    Article  CAS  Google Scholar 

  2. Murali-Krishna, M. et al. Counting antigen-specific CD8 T cells: A reevaluation of bystander activation during viral infection. Immunity 8, 177–188 (1998).

    Article  CAS  Google Scholar 

  3. Manz, R., Assenmacher, M., Pfluger, E., Miltenyi, S. & Radbruch, A. Analysis and sorting of live cells to secreted molecules, relocated to a cell-surface affinity matrix. Proc. Natl. Acad. Sci. USA 92, 1921–1925 (1995).

    Article  CAS  Google Scholar 

  4. Altman, J.D. et al. Phenotypic analysis of antigen specific T lymphocytes. Science 274, 94–96 (1996).

    Article  CAS  Google Scholar 

  5. McMichael, A. & O'Callaghan, C. A new look at T cells. J. Exp. Med. 187, 1367–1371 (1998).

    Article  CAS  Google Scholar 

  6. Busch, D.H., Pilip, I.M., Vijh, S. & Pamer, E.G. Coordinate regulation of complex T cell populations responding to bacterial infection. Immunity 8, 353–362 (1998).

    Article  CAS  Google Scholar 

  7. Matsui, K., Boniface, J.J., Steffner, P., Ray, P.A. & Davis, M.M. Kinetics of T-cell receptor binding to peptide/I-Ek complexes: Correlation of the dissociation rate with T-cell responsiveness. Proc. Natl. Acad. Sci. USA 91, 12862–12866 (1994).

    Article  CAS  Google Scholar 

  8. Busch, D.H. & Pamer, E.G. T cell affinity maturation by selective expansion during infection. J. Exp. Med. 189, 701–709 (1998).

    Article  Google Scholar 

  9. Busch, D.H., Pilip, I. & Pamer, E.G. Evolution of a complex T cell receptor repertoire during primary and recall bacterial infection. J. Exp. Med. 188, 61–70 (1998).

    Article  CAS  Google Scholar 

  10. Novak, E.J., Liu, A.W., Nepom, G.T. & Kwok, W.W. MHC class II tetramers identify peptide-specific human CD4(+) T cells proliferating in response to influenza A antigen. J. Clin. Invest. 104, R63–7 (1999).

    Article  CAS  Google Scholar 

  11. Xu, X.N. et al. A novel approach to antigen-specific deletion of CTL with minimal cellular activation using alpha3 domain mutants of MHC class I/peptide complex. Immunity 14, 591–602 (2001).

    Article  CAS  Google Scholar 

  12. Whelan, J.A. et al. Specificity of CTL interactions with peptide-MHC class I tetrameric complexes is temperature dependent. J. Immunol. 163, 4342–4348 (1999).

    CAS  PubMed  Google Scholar 

  13. Daniels, M.A. & Jameson, S.C. Critical role for CD8 in T cell receptor binding and activation by peptide/major histocompatibility complex multimers. J. Exp. Med. 191, 335–346 (2000).

    Article  CAS  Google Scholar 

  14. Maile, R. et al. Antigen-specific modulation of an immune response by in vivo administration of soluble MHC class I tetramers. J. Immunol. 167, 3708–3714 (2001).

    Article  CAS  Google Scholar 

  15. O'Herrin, S.M. et al. Antigen-specific blockade of T cells in vivo using dimeric MHC peptide. J. Immunol. 167, 2555–2560 (2001).

    Article  CAS  Google Scholar 

  16. O'Reilly, R.J. et al. Biology and adoptive cell therapy of Epstein–Barr virus-associated lymphoproliferative disorders in recipients of marrow allografts. Immunol. Rev. 157, 195–216 (1997).

    Article  CAS  Google Scholar 

  17. Walter, E.A. et al. Reconsitution of cellular immunity against cytomegalovirus in recipients of allogeneic bone marrow by transfer of T-cell clones from the donor. N. Engl. J. Med. 333, 1038–1044 (1995).

    Article  CAS  Google Scholar 

  18. Rooney, C.M. et al. Infusion of cytotoxic T cells for prevention and treatment of Epstein–Barr Virus-induced lymphoma in allogeneic transplant recipients. Blood 92, 1549–1555 (1998).

    CAS  PubMed  Google Scholar 

  19. Busch, D., Kerksiek, K. & Pamer, E. Processing of Listeria monocytogenes antigens and the in vivo T-cell response to bacterial infection. Immunol. Rev. 172, 163–169 (1999).

    Article  CAS  Google Scholar 

  20. Busch, D.H. & Pamer, E.G. MHC class I/peptide stability: Implications for immunodominance, in vitro proliferation and diversity of responding CTL. J. Immunol. 160, 4441–4448 (1998).

    CAS  PubMed  Google Scholar 

  21. Harty, J.T. & Bevan, M.J. CD8 T cells specific for a single nonamer epitope of Listeria monocytogenes are protective in vivo. J. Exp. Med. 175, 1531–1540 (1992).

    Article  CAS  Google Scholar 

  22. Voss, S. & Skerra, A. Mutagenesis of a flexible loop in streptavidin leads to higher affinity for the Strep-tag II peptide and improved performance in recombinant protein purification. Protein Eng. 10, 975–982 (1997).

    Article  CAS  Google Scholar 

  23. Fahmy, T.M., Bieler, J.G., Edidin, M. & Schneck, J.P. Increased TCR avidity after T cell activation: A mechanism for sensing low-density antigen. Immunity 14, 135–143 (2001).

    CAS  PubMed  Google Scholar 

  24. Luxembourg, A.T. et al. Biomagnetic isolation of antigen-specific CD8+ T cells usable in immunotherapy. Nature Biotech. 16, 281–285 (1998).

    Article  CAS  Google Scholar 

  25. Dunbar, P.R. et al. Cutting edge: Rapid cloning of tumor-specific CTL suitable for adoptive immunotherapy of melanoma. J. Immunol. 162, 6959–6962 (1999).

    CAS  PubMed  Google Scholar 

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Acknowledgements

We thank T. Schmidt and G. Holzapfel from IBA for advice during establishment of the MHC-streptagII-StrepTactin technology and for providing StrepTactin polymer reagents; F. Anderl for supportive work during his laboratory rotation, P. Ahmad-Nejad for help with immunohistological studies; and K. Kerksiek, I. Förster and K. Pfeffer for discussions of the manuscript. H.B. is supported by a grant from the German Research Council. D.H.B. is supported by a Gerhard Hess fellowship from the Deutsche Forschungsgemeinschaft. Patent pending.

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  1. Michael Knabel and Tobias J. Franz: M.K & T.J.F. contributed equally to this work

Authors and Affiliations

  1. Institute of Medical Microbiology, Immunology, and Hygiene, Technical University Munich, Munich, Germany

    Michael Knabel, Tobias J. Franz, Matthias Schiemann, Anna Wulf, Brigitte Villmow, Hermann Wagner & Dirk H. Busch

  2. Department of Hematology/Oncology, Technical University of Munich, Munich, Germany

    Burkhard Schmidt & Helga Bernhard

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  1. Michael Knabel
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Correspondence to Dirk H. Busch.

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Knabel, M., Franz, T., Schiemann, M. et al. Reversible MHC multimer staining for functional isolation of T-cell populations and effective adoptive transfer. Nat Med 8, 631–637 (2002). https://doi.org/10.1038/nm0602-631

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