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

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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.

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  • 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.


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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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Corresponding authors

Correspondence to Brian D. Evavold or Cheng Zhu.

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The authors declare no competing financial interests.

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).

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