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Costimulation and endogenous MHC ligands contribute to T cell recognition

Nature Immunology volume 3, pages 42–47 (2002)Cite this article

An Erratum to this article was published on 01 March 2002

Abstract

To initiate an immune response, key receptor-ligand pairs must cluster in “immune synapses” at the T cell–antigen-presenting cell (APC) interface. We visualized the accumulation of a major histocompatibility complex (MHC) class II molecule, I-Ek, at a T cell–B cell interface and found it was dependent on both antigen recognition and costimulation. This suggests that costimulation-driven active transport of T cell surface molecules helps to drive immunological synapse formation. Although only agonist peptide–MHC class II (agonist pMHC class II) complexes can initiate T cell activation, endogenous pMHC class II complexes also appeared to accumulate. To test this directly, we labeled a “null” pMHC class II complex and found that, although it lacked major TCR contact residues, it could be driven into the synapse in a TCR-dependant manner. Thus, low-affinity ligands can contribute to synapse formation and T cell signaling.

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Figure 1: I-Ek accumulates at the T cell–APC interface.
Figure 2: I-Ek accumulation is controlled by costimulation and mediated by TCR but not by CD4 binding.
Figure 3: Null peptide–loaded I-Ek complexes accumulate at the interface.

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Acknowledgements

We thank M. F. Krummel, R. M. Kantor and N. J. Burroughs for helpful discussions. OG-gEk was a gift of E. Hailman. Supported by grants from the Howard Hughes Medical Institute and the National Institutes of Health (to M. M. D.) as well as the Cancer Research Fund of the Damon Runyon Walter Winchell Foundation (L. C. W.).

Author information

Author notes

  1. Michael D. Sjaastad

    Present address: Hyseq Corporation, Sunnyvale, CA, 94085, USA

  2. Michael L. Dustin

    Present address: Department of Pathology, New York University School of Medicine, New York, NY, 10016, USA

Authors and Affiliations

  1. The Howard Hughes Medical Institute and The Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, 94305, CA, USA

    Christoph Wülfing, Cenk Sumen, Lawren C. Wu & Mark M. Davis

  2. Center for Immunology and Department of Cell Biology, UT Southwestern Medical Center, Dallas, 75390, TX, USA

    Christoph Wülfing

  3. Universal Imaging Corporation, Downingtown, 19335, PA, USA

    Michael D. Sjaastad

  4. Department of Pathology and Center for Immunology, Washington University School of Medicine, St. Louis, 63110, MO, USA

    Michael L. Dustin

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Corresponding author

Correspondence to Mark M. Davis.

Additional information

Note: Supplementary information can be found on the Nature Immunology website (http://immunology.nature.com/supp_info/).

Supplementary information

Web Movie 1.

I-Eκ accumulates at the T cell-APC interface: time lapse. The interaction of a 5C.C7 T cell with an A20.I-Eκ cell, peptide loaded with 10 µM agonist peptide, is shown. A bright field series of images has been duplicated and is overlaid with false-color encoded fluorescence information. The top panel is overlaid with a false-color representation of the intracellular calcium concentration of the T cell ranging from blue (low) to red (high calcium concentration). In the bottom panel the I-Eκ-GFP fluorescence of the B cell lymphoma is overlaid in a rainbow color scale from blue (low) to red (high GFP fluorescence). For presentation purposes the blue color has been thresholded out. The I-Eκ-GFP fluorescence has been collected in 21 z-planes, of which only the middle one is shown. The movie shows that I-Eκ-GFP accumulates at the T cell-B cell interface; the accumulation in this particular cell couple is concentrated. The movie compresses 14 min of experiment into 42 s of movie. (MOV 717 kb)

Web Movie 2.

I-Eκ accumulates at the T cell-APC interface: 3D images at selected timepoints. This is derived from the same experiment as Movie 1. A 3D reconstruction of the I-Eκ-GFP fluorescence of the A20.I-Eκ cell interacting with the 5C.C7 T cell from Movie 1 is shown at four timepoints. The position of the T cell-APC interface is visible as a flattening of the bottom left area of the A20 cells. The timepoints from top left to bottom right are: the time of the formation of the T cell-B cell interface and 3 min, 6 min and 9 min afterwards. The I-Eκ-GFP fluorescence color scale is the same as in Movie 1 but without thresholding. The 3D reconstructions are sequentially rotated in 15-degree increments around a horizontal and vertical axis. In this particular experiment the accumulation is concentrated. (MOV 1984 kb)

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Wülfing, C., Sumen, C., Sjaastad, M. et al. Costimulation and endogenous MHC ligands contribute to T cell recognition. Nat Immunol 3, 42–47 (2002). https://doi.org/10.1038/ni741

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