Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

TCRs with high affinity for foreign pMHC show self-reactivity

Abstract

T cells with high-affinity T cell receptors (TCRs) for a foreign peptide–major histocompatibility complex (pMHC) appear to be negatively selected, even though they have never seen the foreign antigen. To examine how this process operates, we used in vitro yeast display to isolate high-affinity TCRs from the T cell clone 2C. The TCRs showed fast on-rates, which were consistent with reduced CDR (complementarity determining region) flexibility, and cross-reactivity with other cognate pMHCs. T cell hybridomas transfected with a high-affinity TCR were stimulated by endogenous self-pMHC, which suggested that T cells bearing the TCR would be negatively selected. The immune system appears to maintain a repertoire of flexible, low-affinity TCRs at the expense of more effective high-affinity TCRs.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Binding of pMHC complexes to yeast-displayed CDR3α mutants.
Figure 2: Sequences of CDR3α regions from high-affinity TCRs.
Figure 3: Binding of soluble TCRs to pMHC complexes on cells.
Figure 4: Functional activity of mutant m33α and WT 2C TCRs transfected into the T cell hybridoma 58−/−.
Figure 5: Binding assays of mutant TCRs and SIYR-Kb.
Figure 6: Energy-minimized models of high-affinity TCR-pMHC complexes.

Similar content being viewed by others

Accession codes

Accessions

Protein Data Bank

References

  1. Davis, M.M. & Bjorkman, P.J. T-cell antigen receptor genes and T-cell recognition. Nature 334, 395–402 (1988).

    CAS  PubMed  Google Scholar 

  2. Goldrath, A.W. & Bevan, M.J. Selecting and maintaining a diverse T-cell repertoire. Nature 402, 255–262 (1999).

    CAS  PubMed  Google Scholar 

  3. Sherman, L.A. et al. Self-tolerance and the composition of T cell repertoire. Immunol. Res. 21, 305–313 (2000).

    CAS  PubMed  Google Scholar 

  4. Zerrahn, J., Held, W. & Raulet, D.H. The MHC reactivity of the T cell repertoire prior to positive and negative selection. Cell 88, 627–636 (1997).

    CAS  PubMed  Google Scholar 

  5. Alam, S.M. et al. T cell receptor affinity and thymocyte positive selection. Nature 381, 616–620 (1996).

    CAS  Google Scholar 

  6. Evavold, B.D., Sloan-Lancaster, J., Wilson, K.J., Rothbard, J.B. & Allen, P.M. Specific T cell recognition of minimally homologous peptides: Evidence for multiple endogenous ligands. Immunity 2, 655–663 (1995).

    CAS  PubMed  Google Scholar 

  7. Mason, D. A very high level of crossreactivity is an essential feature of the T-cell receptor. Immunol. Today 19, 395–404 (1998).

    CAS  PubMed  Google Scholar 

  8. Garcia, K.C. et al. Structural basis of plasticity in T cell receptor recognition of a self peptide-MHC antigen. Science 279, 1166–1172 (1998).

    CAS  PubMed  Google Scholar 

  9. Reiser, J.B. et al. A T cell receptor CDR3β loop undergoes conformational changes of unprecedented magnitude upon binding to a peptide/MHC class I complex. Immunity 16, 345–354 (2002).

    CAS  PubMed  Google Scholar 

  10. Hare, B.J. et al. Structure, specificity and CDR mobility of a class II restricted single-chain T-cell receptor. Nat. Struct. Biol. 6, 574–581 (1999).

    CAS  PubMed  Google Scholar 

  11. Garcia, K.C., Degano, M., Speir, J.A. & Wilson, I.A. Emerging principles for T cell receptor recognition of antigen in cellular immunity. Rev. Immunogenet. 1, 75–90 (1999).

    CAS  PubMed  Google Scholar 

  12. Willcox, B.E. et al. TCR binding to peptide-MHC stabilizes a flexible recognition interface. Immunity 10, 357–365 (1999).

    CAS  PubMed  Google Scholar 

  13. Boniface, J.J., Reich, Z., Lyons, D.S. & Davis, M.M. Thermodynamics of T cell receptor binding to peptide-MHC: Evidence for a general mechanism of molecular scanning. Proc. Natl. Acad. Sci. USA 96, 11446–11451 (1999).

    CAS  PubMed  Google Scholar 

  14. van der Merwe, P.A. The TCR triggering puzzle. Immunity 14, 665–668 (2001).

    CAS  PubMed  Google Scholar 

  15. Savage, P.A., Boniface, J.J. & Davis, M.M. A kinetic basis for T cell receptor repertoire selection during an immune response. Immunity 10, 485–492 (1999).

    CAS  PubMed  Google Scholar 

  16. Savage, P.A. & Davis, M.M. A kinetic window constricts the T cell receptor repertoire in the thymus. Immunity 14, 243–252 (2001).

    CAS  PubMed  Google Scholar 

  17. Holler, P.D., Lim, A.R., Cho, B.K., Rund, L.A. & Kranz, D.M. CD8 T cell transfectants that express a high affinity T cell receptor exhibit enhanced peptide-dependent activation. J. Exp. Med. 194, 1043–1052 (2001).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Holler, P.D. et al. In vitro evolution of a T cell receptor with high affinity for peptide/MHC. Proc. Natl. Acad. Sci. USA 97, 5387–5392 (2000).

    CAS  PubMed  Google Scholar 

  19. Boder, E.T. & Wittrup, K.D. Yeast surface display for screening combinatorial polypeptide libraries. Nat. Biotech. 15, 553–557 (1997).

    CAS  Google Scholar 

  20. Kieke, M.C., Cho, B.K., Boder, E.T., Kranz, D.M. & Wittrup, K.D. Isolation of anti-T cell receptor scFv mutants by yeast surface display. Protein Engineer. 10, 1303–1310 (1997).

    CAS  Google Scholar 

  21. Shusta, E.V., Holler, P.D., Kieke, M.C., Kranz, D.M. & Wittrup, K.D. Directed evolution of a stable scaffold for T-cell receptor engineering. Nat. Biotechnol. 18, 754–759 (2000).

    CAS  PubMed  Google Scholar 

  22. Kieke, M.C. et al. Selection of functional T cell receptor mutants from a yeast surface-display library. Proc. Natl. Acad. Sci. USA 96, 5651–5656 (1999).

    CAS  PubMed  Google Scholar 

  23. Garcia, K.C. et al. αβ T cell receptor interactions with syngeneic and allogeneic ligands: affinity measurements and crystallization. Proc. Natl. Acad. Sci. USA 94, 13838–13843 (1997).

    CAS  PubMed  Google Scholar 

  24. Speir, J.A. et al. Structural basis of the 2C TCR allorecognition of H-2Ld peptide complexes. Immunity 8, 553–562 (1998).

    CAS  PubMed  Google Scholar 

  25. Schlueter, C.J., Manning, T.C., Schodin, B.A. & Kranz, D.M. A residue in the center of peptide QL9 affects binding to both Ld and the T cell receptor. J. Immunol. 157, 4478–4485 (1996).

    CAS  PubMed  Google Scholar 

  26. Schodin, B.A., Tsomides, T.J. & Kranz, D.M. Correlation between the number of T cell receptors required for T cell activation and TCR-ligand affinity. Immunity 5, 137–146 (1996).

    CAS  PubMed  Google Scholar 

  27. Manning, T.C., Schodin, B.A. & Kranz, D.M. A strategy for the synthesis and screening of thiol-modified peptide variants recognized by T cells. J. Immunol. Meth. 192, 125–132 (1996).

    CAS  Google Scholar 

  28. Davis, M.M. et al. Ligand recognition by αβ T cell receptors. Annu. Rev. Immunol. 16, 523–544 (1998).

    CAS  PubMed  Google Scholar 

  29. Gascoigne, N.R.J., Zai, T. & Alam, S.M. T-cell receptor binding kinetics in T-cell development and activation. Exp. Rev. Mol. Med. (12 February 2001), http://www-ermm.cbcu.cam.ac.uk/01002502h.htm.

  30. Sha, W.C. et al. Positive selection of transgenic receptor-bearing thymocytes by Kb antigen is altered by Kb mutations that involve peptide binding. Proc. Natl. Acad. Sci. USA 87, 6186–6191 (1990).

    CAS  PubMed  Google Scholar 

  31. Manning, T.C., Parke, E.A., Teyton, L. & Kranz, D.M. Effects of complementarity determining region mutations on the affinity of an αβ T cell receptor: measuring the energy associated with CD4/CD8 repertoire skewing. J. Exp. Med. 189, 461–470 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Lee, P.U., Churchill, H.R., Daniels, M., Jameson, S.C. & Kranz, D.M. Role of 2C T cell receptor residues in the binding of self- and allo-major histocompatibility complexes. J. Exp. Med. 191, 1355–1364 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Eisen, H.N. Specificity and degeneracy in antigen recognition: yin and yang in the immune system. Annu. Rev. Immunol. 19, 1–21 (2001).

    CAS  PubMed  Google Scholar 

  34. Degano, M. et al. A functional hot spot for antigen recognition in a superagonist TCR/MHC complex. Immunity 12, 251–261 (2000).

    CAS  PubMed  Google Scholar 

  35. Ostrov, D.A., Shi, W., Schwartz, J.C., Almo, S.C. & Nathenson, S.G. Structure of murine CTLA-4 and its role in modulating T cell responsiveness. Science 290, 816–819 (2000).

    CAS  PubMed  Google Scholar 

  36. Manivel, V., Sahoo, N.C., Salunke, D.M. & Rao, K.V. Maturation of an antibody response is governed by modulations in flexibility of the antigen-combining site. Immunity 13, 611–620 (2000).

    CAS  PubMed  Google Scholar 

  37. Eisen, H.N., Sykulev, Y. & Tsomides, T.J. Antigen-specific T-cell receptors and their reactions with complexes formed by peptides with major histocompatibility complex (MHC) proteins. Adv. Protein Chem. 49, 1–56 (1996).

    CAS  PubMed  Google Scholar 

  38. Valitutti, S., Muller, S., Cella, M., Padovan, E. & Lanzavecchia, A. Serial triggering of many T-cell receptors by a few peptide-MHC complexes. Nature 375, 148–151 (1995).

    CAS  PubMed  Google Scholar 

  39. Kalergis, A.M. et al. Efficient T cell activation requires an optimal dwell-time of interaction between the TCR and the pMHC complex. Nat. Immunol. 2, 229–234 (2001).

    CAS  PubMed  Google Scholar 

  40. Andersen, P.S., Geisler, C., Buus, S., Mariuzza, R.A. & Karjalainen, K. Role of TCR-ligand affinity in T cell activation by bacterial superantigens. J. Biol. Chem. 276, 33452–33457 (2001).

    CAS  PubMed  Google Scholar 

  41. Bouneaud, C., Kourilsky, P. & Bousso, P. Impact of negative selection on the T cell repertoire reactive to a self-peptide: a large fraction of T cell clones escapes clonal deletion. Immunity 13, 829–840 (2000).

    CAS  PubMed  Google Scholar 

  42. Correia-Neves, M., Waltzinger, C., Mathis, D. & Benoist, C. The shaping of the T cell repertoire. Immunity 14, 21–32 (2001).

    CAS  PubMed  Google Scholar 

  43. O'Herrin, S.M. et al. Analysis of the expression of peptide-major histocompatibility complexes using high affinity soluble divalent T cell receptors. J. Exp. Med. 186, 1333–1345 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Garboczi, D.N. et al. Assembly, specific binding, and crystallization of a human TCR-αβ with an antigenic Tax peptide from human T lymphotropic virus type 1 and the class I MHC molecule HLA-A2. J. Immunol. 157, 5403–5410 (1996).

    CAS  PubMed  Google Scholar 

  45. Geitz, R.D., Schiestl, R.H., Willems, A. & Woods, R.A. Studies on the mechanism of high efficiency transformations of intact yeast cells. Yeast 11, 355–360 (1995).

    Google Scholar 

  46. Alexander, J., Payne, J.A., Murray, R., Frelinger, J.A. & Cresswell, P. Differential transport requirements of HLA and H-2 class I glycoproteins. Immunogenetics 29, 380–399 (1989).

    CAS  PubMed  Google Scholar 

  47. Kranz, D.M., Sherman, D.H., Sitkovsky, M.V., Pasternack, M.S. & Eisen, H.N. Immunoprecipitation of cell surface structure of cloned cytotoxic T lymphocytes by clone-specific antisera. Proc. Natl. Acad. Sci. USA 81, 573–577 (1984).

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank S. Jameson and M. Daniels for the Kb plasmid and advice on tetramer preparation; the NIH Tetramer facility for several of the initial preparations of pMHC tetramers; I. Wilson and K.D. Wittrup for helpful discussions; and G. Durack and the Biotech Flow Facility for expert technical advice and assistance. Supported by NIH Grant RO1 GM55767.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to David M. Kranz.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Holler, P., Chlewicki, L. & Kranz, D. TCRs with high affinity for foreign pMHC show self-reactivity. Nat Immunol 4, 55–62 (2003). https://doi.org/10.1038/ni863

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni863

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing