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New insights into the T cell synapse from single molecule techniques

Nature Reviews Immunology volume 11, pages 672–684 (2011)Cite this article

Subjects

Key Points

  • Single molecule methods are leading to advances in our understanding of immune cell activation, but different interpretations of the cutting-edge data can also trigger controversies.

  • Probing single T cell receptor (TCR)–peptide–MHC bonds at early time points has led to the discovery that the two-dimensional on-rate is a crucial parameter that amplifies apparently small chemical differences between different ligand–receptor interactions.

  • Measuring single TCR–peptide–MHC interactions using fluorescence resonance energy transfer reveals that cytoskeletal force increases the off-rate by approximately tenfold, further emphasizing the importance of efficient peptide–MHC capture.

  • Electron microscopy and super-resolution fluorescence microscopy reveal that the TCR and the adaptor protein linker for activation of T cells (LAT) are organized into protein islands in a protein-poor lipid sea and that collisions between TCR+ and LAT+ islands initiate signalling.

  • By contrast, time-lapse and super-resolution fluorescence microscopy suggest that TCR clusters in the plasma membrane interact in trans with LAT in sub-synaptic vesicles to initiate signalling.

  • Mutations in a putative TCR dimer interface impair central supramolecular activation cluster (cSMAC) formation, a TSG101-dependent process that might transfer nucleic acid messages from T cells to antigen-presenting cells.

Abstract

T cell activation depends on extracellular ligation of the T cell receptor (TCR) by peptide–MHC complexes in a synapse between the T cell and an antigen-presenting cell. The process then requires the assembly of signalling complexes between the TCR and the adaptor protein linker for activation of T cells (LAT), and subsequent filamentous actin (F-actin)-dependent TCR cluster formation. Recent progress in each of these areas, made possible by the emergence of new techniques, has forced us to rethink our assumptions and consider some radical new models. These describe the receptor interaction parameters that control T cell responses and the mechanism by which LAT is recruited to the TCR signalling machinery. This is an exciting time in T cell biology, and further innovation in imaging and genomics is likely to lead to a greater understanding of how T cells are activated.

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Figure 1: Structure of the immunological synapse.
Figure 2: TCR dimerization.
Figure 3: Two views of TCR+ and LAT+ clusters.
Figure 4: Model for how F-actin can promote a fast TCR on-rate.

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Acknowledgements

We thank R. Varma, K. Choudhuri, S. Kumari and N. Destainville for helpful discussions. This work was supported by US National Institutes of Health grants R01 AI043549, P01 AI045757 and PN2 EY016586.

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  1. Department of Pathology, Helene and Martin Kimmel Center for Biology and Medicine of the Skirball Institute of Biomolecular Medicine, New York University School of Medicine, 540 First Avenue, New York, 10012, New York, USA

    Michael L. Dustin & David Depoil

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Correspondence to Michael L. Dustin.

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Glossary

Supramolecular activation clusters

(SMACs). During T cell activation, TCRs accumulate into a central cluster termed the central SMAC (cSMAC) at the interface between the T cell and the antigen-presenting cell. The cSMAC is surrounded by an LFA1 ring termed the peripheral SMAC (pSMAC) in a bulls-eye manner, and this characteristic receptor organization (the cSMAC surrounded by the pSMAC) constitutes the mature immunological synapse. Large proteins are excluded from the mature immunological synapse and form the distal SMAC (dSMAC).

Immunoreceptor tyrosine-based activation motifs

(ITAMs). The ITAM is an amino acid sequence — Yxx(I/L)x6–12Yxx(I/L) — found in a large number of receptors and adaptor proteins. After phosphorylation, ITAMs function as docking sites for proteins that contain tandem SH2 domains, such as ZAP70.

ESCRT proteins

(Endosomal sorting complex required for transport proteins). The ESCRT proteins coordinate the degradation of ubiquitylated substrates through multivesicular bodies. There are four ESCRT complexes (0, I, II and III) with unique roles in signal termination and receptor degradation.

Polarity network

Evolutionarily conserved cytoplasmic proteins that have been defined using genetic screens for gene products that are required for cell polarity (for example, apical versus basal and front versus back).

Sub-synaptic vesicles

Endosomal membranes and other vesicular structures less than 200 nm below the plasma membrane in the immunological synapse.

Lipid rafts

Lipid domains that are enriched in cholesterol, sphingolipids and phospholipids with saturated acyl chains. The lipid phase of these regions is referred to as 'liquid ordered', in contrast to the liquid-disordered domains that characterize the bulk of biological membranes. Lipid rafts are small (∼70 nm across) and dynamic.

Cytoskeletal corrals

Membrane regions surrounded by barriers that are formed by proteins associated with the membrane cytoskeleton. These barriers limit the diffusion of proteins within the plasma membrane.

Cortical filament

A fibre composed of actin and other cytoskeletal proteins that is present in close proximity to the cell membrane. These filaments influence membrane geometry and the behaviour of proteins in the membrane.

Actin rocket

Actin polymerization induced by a particle such as a virus, bacterium or vesicle can propel the object in the cytoplasm with a tail of polymerized actin that is reminiscent of a rocket.

Glycocalyx

An extracellular polymer composed of glycoproteins that covers the outside of eukaryotic cells.

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Dustin, M., Depoil, D. New insights into the T cell synapse from single molecule techniques. Nat Rev Immunol 11, 672–684 (2011). https://doi.org/10.1038/nri3066

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