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The kinetics of two-dimensional TCR and pMHC interactions determine T-cell responsiveness

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Abstract

The T-cell receptor (TCR) interacts with peptide-major histocompatibility complexes (pMHC) to discriminate pathogens from self-antigens and trigger adaptive immune responses. Direct physical contact is required between the T cell and the antigen-presenting cell for cross-junctional binding where the TCR and pMHC are anchored on two-dimensional (2D) membranes of the apposing cells1. Despite their 2D nature, TCR–pMHC binding kinetics have only been analysed three-dimensionally (3D) with a varying degree of correlation with the T-cell responsiveness2,3,4. Here we use two mechanical assays5,6 to show high 2D affinities between a TCR and its antigenic pMHC driven by rapid on-rates. Compared to their 3D counterparts, 2D affinities and on-rates of the TCR for a panel of pMHC ligands possess far broader dynamic ranges that match that of their corresponding T-cell responses. The best 3D predictor of response is the off-rate, with agonist pMHC dissociating the slowest2,3,4. In contrast, 2D off-rates are up to 8,300-fold faster, with the agonist pMHC dissociating the fastest. Our 2D data suggest rapid antigen sampling by T cells and serial engagement of a few agonist pMHCs by TCRs in a large self pMHC background. Thus, the cellular environment amplifies the intrinsic TCR–pMHC binding to generate broad affinities and rapid kinetics that determine T-cell responsiveness.

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Figure 1: Micropipette and BFP.
Figure 2: 2D kinetics measurements.
Figure 3: Comparison of 2D and 3D kinetics.
Figure 4: Correlation between 2D kinetics and T-cell proliferation.

Change history

  • 12 April 2010

    In the online–only Methods, Pa was changed to Passoc in three places on 12 April 2010. Please see the erratum at the end of the PDF for details.

References

  1. 1

    Dustin, M. L., Bromley, S. K., Davis, M. M. & Zhu, C. Identification of self through two-dimensional chemistry and synapses. Annu. Rev. Cell Dev. Biol. 17, 133–157 (2001)

    CAS  Article  Google Scholar 

  2. 2

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

    ADS  CAS  Article  Google Scholar 

  3. 3

    Gascoigne, N. R., Zal, T. & Alam, S. M. T-cell receptor binding kinetics in T-cell development and activation. Expert Rev. Mol. Med. 2001, 1–17 (2001)

    CAS  Article  Google Scholar 

  4. 4

    Stone, J. D., Chervin, A. S. & Kranz, D. M. T-cell receptor binding affinities and kinetics: impact on T-cell activity and specificity. Immunology 126, 165–176 (2009)

    CAS  Article  Google Scholar 

  5. 5

    Chesla, S. E., Selvaraj, P. & Zhu, C. Measuring two-dimensional receptor-ligand binding kinetics by micropipette. Biophys. J. 75, 1553–1572 (1998)

    CAS  Article  Google Scholar 

  6. 6

    Chen, W., Evans, E. A., McEver, R. P. & Zhu, C. Monitoring receptor-ligand interactions between surfaces by thermal fluctuations. Biophys. J. 94, 694–701 (2008)

    CAS  Article  Google Scholar 

  7. 7

    Huang, J., Edwards, L. J., Evavold, B. D. & Zhu, C. Kinetics of MHC-CD8 interaction at the T cell membrane. J. Immunol. 179, 7653–7662 (2007)

    CAS  Article  Google Scholar 

  8. 8

    Chen, W., Zarnitsyna, V. I., Sarangapani, K. K., Huang, J. & Zhu, C. Measuring receptor-ligand binding kinetics on cell surfaces: from adhesion frequency to thermal fluctuation methods. Cell. Mol. Bioeng. 1, 276–288 (2008)

    CAS  Article  Google Scholar 

  9. 9

    Zarnitsyna, V. I. et al. Memory in receptor-ligand-mediated cell adhesion. Proc. Natl Acad. Sci. USA 104, 18037–18042 (2007)

    ADS  CAS  Article  Google Scholar 

  10. 10

    Howarth, M. et al. A monovalent streptavidin with a single femtomolar biotin binding site. Nature Methods 3, 267–273 (2006)

    CAS  Article  Google Scholar 

  11. 11

    Alam, S. M. et al. Qualitative and quantitative differences in T cell receptor binding of agonist and antagonist ligands. Immunity 10, 227–237 (1999)

    CAS  Article  Google Scholar 

  12. 12

    Rosette, C. et al. The impact of duration versus extent of TCR occupancy on T cell activation: a revision of the kinetic proofreading model. Immunity 15, 59–70 (2001)

    CAS  Article  Google Scholar 

  13. 13

    Zhang, F. et al. Two-dimensional kinetics regulation of αLβ2-ICAM-1 interaction by conformational changes of the αL-inserted domain. J. Biol. Chem. 280, 42207–42218 (2005)

    CAS  Article  Google Scholar 

  14. 14

    Huang, J. et al. Quantifying the effects of molecular orientation and length on two-dimensional receptor-ligand binding kinetics. J. Biol. Chem. 279, 44915–44923 (2004)

    CAS  Article  Google Scholar 

  15. 15

    Mehta, P., Cummings, R. D. & McEver, R. P. Affinity and kinetic analysis of P-selectin binding to P-selectin glycoprotein ligand-1. J. Biol. Chem. 273, 32506–32513 (1998)

    CAS  Article  Google Scholar 

  16. 16

    Wu, L. C., Tuot, D. S., Lyons, D. S., Garcia, K. C. & Davis, M. M. Two-step binding mechanism for T-cell receptor recognition of peptide MHC. Nature 418, 552–556 (2002)

    ADS  CAS  Article  Google Scholar 

  17. 17

    Nicholson, M. W., Barclay, A. N., Singer, M. S., Rosen, S. D. & van der Merwe, P. A. Affinity and kinetic analysis of L-selectin (CD62L) binding to glycosylation-dependent cell-adhesion molecule-1. J. Biol. Chem. 273, 763–770 (1998)

    CAS  Article  Google Scholar 

  18. 18

    Zehn, D., Lee, S. Y. & Bevan, M. J. Complete but curtailed T-cell response to very low-affinity antigen. Nature 458, 211–214 (2009)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Schamel, W. W. et al. Coexistence of multivalent and monovalent TCRs explains high sensitivity and wide range of response. J. Exp. Med. 202, 493–503 (2005)

    CAS  Article  Google Scholar 

  20. 20

    Lillemeier, B. F. et al. TCR and Lat are expressed on separate protein islands on T cell membranes and concatenate during activation. Nature Immunol. 11, 90–96 (2009)

    Article  Google Scholar 

  21. 21

    Campi, G., Varma, R. & Dustin, M. L. Actin and agonist MHC-peptide complex-dependent T cell receptor microclusters as scaffolds for signaling. J. Exp. Med. 202, 1031–1036 (2005)

    CAS  Article  Google Scholar 

  22. 22

    Yokosuka, T. et al. Newly generated T cell receptor microclusters initiate and sustain T cell activation by recruitment of Zap70 and SLP-76. Nature Immunol. 6, 1253–1262 (2005)

    CAS  Article  Google Scholar 

  23. 23

    Zidovetzki, R. & Levitan, I. Use of cyclodextrins to manipulate plasma membrane cholesterol content: evidence, misconceptions and control strategies. Biochim. Biophys. Acta 1768, 1311–1324 (2007)

    CAS  Article  Google Scholar 

  24. 24

    Aleksic, M. O. et al. Dependence of T cell antigen recognition on T cell receptor-peptide MHC confinement time. Immunity 32, 163–174 (2010)

    CAS  Article  Google Scholar 

  25. 25

    Davis, S. J. & van der Merwe, P. A. The kinetic-segregation model: TCR triggering and beyond. Nature Immunol. 7, 803–809 (2006)

    CAS  Article  Google Scholar 

  26. 26

    McKeithan, T. W. Kinetic proofreading in T-cell receptor signal transduction. Proc. Natl Acad. Sci. USA 92, 5042–5046 (1995)

    ADS  CAS  Article  Google Scholar 

  27. 27

    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)

    ADS  CAS  Article  Google Scholar 

  28. 28

    Daniels, M. A. et al. Thymic selection threshold defined by compartmentalization of Ras/MAPK signalling. Nature 444, 724–729 (2006)

    ADS  CAS  Article  Google Scholar 

  29. 29

    Davis, M. M. et al. T cells as a self-referential, sensory organ. Annu. Rev. Immunol. 25, 681–695 (2007)

    CAS  Article  Google Scholar 

  30. 30

    Henrickson, S. E. et al. T cell sensing of antigen dose governs interactive behavior with dendritic cells and sets a threshold for T cell activation. Nature Immunol. 9, 282–291 (2008)

    CAS  Article  Google Scholar 

  31. 31

    Sabatino, J. J., Shires, J., Altman, J. D., Ford, M. L. & Evavold, B. D. Loss of IFN-γ enables the expansion of autoreactive CD4+ T cells to induce experimental autoimmune encephalomyelitis by a nonencephalitogenic myelin variant antigen. J. Immunol. 180, 4451–4457 (2008)

    CAS  Article  Google Scholar 

  32. 32

    Evans, E., Leung, A., Heinrich, V. & Zhu, C. Mechanical switching and coupling between two dissociation pathways in a P-selectin adhesion bond. Proc. Natl Acad. Sci. USA 101, 11281–11286 (2004)

    ADS  CAS  Article  Google Scholar 

  33. 33

    Hochmuth, R. M. in Handbook of Bioengineering (eds Skalak, R. & Chien, S.) (McGraw-Hill, 1987)

    Google Scholar 

  34. 34

    Lou, J. et al. Flow-enhanced adhesion regulated by a selectin interdomain hinge. J. Cell Biol. 174, 1107–1117 (2006)

    CAS  Article  Google Scholar 

  35. 35

    Zhu, C., Long, M. & Bongrand, P. Measuring receptor/ligand interaction at the single bond level: experimental and interpretive issues. Ann. Biomed. Eng. 30, 305–314 (2002)

    Article  Google Scholar 

  36. 36

    Williams, T. E., Selvaraj, P. & Zhu, C. Concurrent binding to multiple ligands: kinetic rates of CD16b for membrane-bound IgG1 and IgG2. Biophys. J. 79, 1858–1866 (2000)

    CAS  Article  Google Scholar 

  37. 37

    Williams, T. E., Nagarajan, S., Selvaraj, P. & Zhu, C. Quantifying the impact of membrane microtopology on effective two-dimensional affinity. J. Biol. Chem. 276, 13283–13288 (2001)

    CAS  Article  Google Scholar 

  38. 38

    Wu, L. et al. Impact of carrier stiffness and microtopology on two-dimensional kinetics of P-selectin and P-selectin glycoprotein ligand-1 (PSGL-1) interactions. J. Biol. Chem. 282, 9846–9854 (2007)

    CAS  Article  Google Scholar 

  39. 39

    Evans, E., Heinrich, V., Leung, A. & Kinoshita, K. Nano- to microscale dynamics of P-selectin detachment from leukocyte interfaces. I. Membrane separation from the cytoskeleton. Biophys. J. 88, 2288–2298 (2005)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank the NIH Tetramer Core Facility at Emory University for providing MHC monomers, J. Altman for providing the H-2Kb mutant construct, G. Bao for providing the divalent streptavidin, as well as M. Davis and H.-T. He for commenting on the manuscript. This work was supported by NIH grants AI38282 and AI060799 (to C.Z.) and NS062358, and National Multiple Sclerosis Society Grant RG4047-A-3 (to B.D.E.).

Author Contributions N.J. and C.Z. initiated the research with F5 T cells; J.H. and C.Z. designed the kinetic study; J.H. and B.L. performed the micropipette experiments; V.I.Z. performed the BFP experiments and Monte Carlo simulations; L.J.E. and B.D.E. provided the T cells and conducted the functional study; J.H., V.I.Z., B.L. and C.Z. analysed the data; B.D.E. and C.Z. wrote the paper with all authors contributing.

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Correspondence to Brian D. Evavold or Cheng Zhu.

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Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-12 with legends and Supplementary Movie legends. (PDF 1058 kb)

Supplementary Movie 1

In this movie we see micropipette adhesion frequency assay (see Supplementary Information file for full legend). (MOV 3871 kb)

Supplementary Movie 2

In this movie we see BFP adhesion frequency assay (see Supplementary Information file for full legend). (MOV 4566 kb)

Supplementary Movie 3

In this movie we see BFP thermal fluctuation assay (see Supplementary Information file for full legend). (MOV 267 kb)

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Huang, J., Zarnitsyna, V., Liu, B. et al. The kinetics of two-dimensional TCR and pMHC interactions determine T-cell responsiveness. Nature 464, 932–936 (2010). https://doi.org/10.1038/nature08944

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