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Surface-bound chemokines capture and prime T cells for synapse formation

A Corrigendum to this article was published on 01 November 2006

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Abstract

T cell activation in vivo occurs in a lymphoid milieu that presents chemotactic and T cell receptor signals concurrently. Here we demonstrate that T cell zone chemokines such as CCL21 are bound to the surface of lymph node dendritic cells. Contact with antigen-presenting cells bearing chemokines costimulated T cells by a previously unknown two-step contact mechanism. T cells initially formed an antigen-independent 'tethered' adhesion on chemokine-bearing antigen-presenting cells. The formation of those tethers superseded T cell receptor signaling and immunological synapse formation. However, chemokine-tethered T cells were hyper-responsive to subsequent contacts with antigen-presenting cells. Thus, T cells are costimulated 'in trans' and sequentially after initial engagement with their chemokine-rich environment.

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Figure 1: CCL21 is stably bound to the surface of lymph node–derived DCs.
Figure 2: Surface-bound chemokines induce antigen-independent tethered adhesion of T cells to APCs.
Figure 3: CCL21 mediates a two-step dynamic of T cell activation with a tethered intermediate that leads to augmentation of antigen-specific coupling.
Figure 4: Surface-bound CCL21 presented in trans is sufficient to augment antigen-specific coupling of a T cell to a separate APC.
Figure 5: Chemokine-induced tethers are dependent on LFA-1 and ICAM but do not permit TCR-mediated signaling.
Figure 6: Naive T cells are captured by APCs in the presence of CCL21, resulting in improved antigen-specific coupling.

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Change history

  • 29 September 2006

    Along the horizontal axis for Figure 5a, anti-LFA-1 at the far right should read anti-β1 (α-β1

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Acknowledgements

We thank J. Cyster, E. Brown and T. Okada for insight and help with reagents; C. Bennett for technical support; S.W. Jiang for flow cytometry; and J. Cyster, S. Rosen and R. Locksley for critical reading of the manuscript.

Author information

Authors and Affiliations

Authors

Contributions

R.S.F. performed all experiments and data analysis and participated in experimental design and manuscript writing. J.J. assisted with experimental design, some experiments and manuscript writing, provided intellectual guidance and performed blinded quantifications. M.F.K. established the initial scientific questions, provided continuing intellectual guidance and participated in experimental design and manuscript writing.

Corresponding author

Correspondence to Matthew F Krummel.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

CCL21-induced costimulation of naive T cell proliferation is blocked by Pertussis toxin. (PDF 1541 kb)

Supplementary Fig. 2

Model: Tethering to chemokine-bearing cells allows T cells to pause and survey their environment, and primes T cells for antigen specific coupling and proliferation. (PDF 2932 kb)

Supplementary Video 1

Prototypical antigen dependent coupling. Representative image of T cell immunological synapse on an antigen-bearing APC as quantified in Fig. 3b. Time-lapse series of DIC (left) and Fura-2AM calcium (right) images of labeled T cell blasts interacting with A20 B cell APCs in the presence of 1μg/ml OVA peptide. The T cell contacts the APC with its leading edge, fluxes calcium and shows the canonical IS morphology – rounded with a flat interface. Calcium flux is on a pseudocolor scale indicating the ratio of Fura-2AM emissions at 340nm/380nm. A timestamp (bottom) indicates the elapsed time in minutes:seconds. (MOV 1702 kb)

Supplementary Video 2

CCL21 induced T cell tethering. Movie version of Fig. 2a. Representative image of T cell tether on a chemokine-bearing APC as quantified in Fig. 2b. A timestamp (bottom) indicates the elapsed time. DIC (left) and corresponding FURA-2AM ratio (right) are shown. (MOV 1176 kb)

Supplementary Video 3

CCL21 induced two-step T cell activation with a tethered intermediate on an antigen-bearing APC. Movie version of Figure 3a. Representative image of T cell tethering followed by immunological synapse formation on an antigen and chemokine-bearing APC as quantified in Fig. 3b. A timestamp (bottom) indicates the elapsed time. DIC (left) and corresponding FURA-2AM ratio (right) are shown. (MOV 2386 kb)

Supplementary Video 4

Two-step T cell activation induced by endogenously produced chemokine on antigen bearing DCs. Timelapse series of DIC (left) and Fura-2AM calcium (right) images of labeled T cell blasts interacting with bone-marrow derived DCs in the presence of 1μg/ml OVA peptide. The T cell contacts the APC with its leading edge, crawls along the APC, and remains attached or tethered to the APC via its uropod. The tethered T cell maintains its polarized amoeboid morphology with an active leading edge and constricted uropod and does not flux calcium until a second contact with a DC induces calcium flux and immunological synapse formation. The cells two of interest are indicated by white and yellow the arrows. Calcium flux is on a pseudocolor scale indicating the ratio of Fura-2AM emissions at 340nm/380nm. A timestamp (top) indicates the elapsed time in minutes:seconds. (MOV 1876 kb)

Supplementary Video 5

CCL21 induced tethering in naïve T cells. Representative image of T cell tether on a chemokine-bearing APC as quantified in Fig. 6a. Timelapse series of DIC (left) and Fura-2AM calcium (right) images of labeled naïve DO11.10 T cell interacting with bone-marrow derived DCs in the presence of 1μg/ml CCL21. The T cell contact the APC and remain bound to it. While bound, the T cells fluctuate between a rounded morphology and a polarized morphology characteristic of a tethered T cell blast. Calcium flux is on a pseudocolor scale indicating the ratio of Fura-2AM emissions at 340nm/380nm. A timestamp (top) indicates the elapsed time in minutes:seconds. (MOV 2024 kb)

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Friedman, R., Jacobelli, J. & Krummel, M. Surface-bound chemokines capture and prime T cells for synapse formation. Nat Immunol 7, 1101–1108 (2006). https://doi.org/10.1038/ni1384

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