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Super-resolution characterization of TCR-dependent signaling clusters.

Sherman Eilon, Barr Valarie, Samelson Lawrence E

📰 Immunological reviews 📅 2013 📊 70 citations

Abstract

SummaryMulti‐molecular signaling complexes drive the earliest events of immune cell activation via immunoreceptors with unexplained specificity and speed. Fluorescence microscopy has shown that these complexes form microclusters at the plasma membrane of activated T cells upon engagement of their antigen receptors (TCRs). Although crucial for cell function, much remains to be learned about the molecular content, fine structure, formation mechanisms, and function of these microclusters. Recent advancements in super‐resolution microscopy have enabled the study of signaling microclusters at the single molecule level with resolution down to approximately 20 nm. These techniques have now helped to characterize the size distributions of signaling clusters at the plasma membrane of intact cells and to shed light on the formation mechanisms that govern their assembly. Surprisingly, dynamic and functional nanostructures have been identified within the signaling clusters. We expect that these novel methodologies, combined with older techniques, will shed new light on the nature of signaling clusters and their critical role in T‐cell activation.

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📊 Figures

Fig. 1

Signaling complexes drive early T-cell activation

T-cell activation is initiated by the T-cell antigen receptor recognition of its peptide-MHC ligand on the surface of antigen-presenting cells (APCs). Following antigen recognition and costimulatory s...

Fig. 2

Signaling complexes reside in clusters smaller than the diffraction limit of light

Confocal images of Jurkat E6.1 cells spread on (A) TCR stimulating (u03b1CD3-coated) and (B) TCR non-stimulating (u03b1CD45-coated) coverslips. The cells were stained with primary proteins against pho...

Fig. 3

A `unified view' of molecular clustering at the PM

We propose a model in which some membrane molecules such as the TCR and LAT cluster in a broad distribution of sizes. The distribution is strongly skewed towards monomers and very small clusters, e.g....

Fig. 4

PALM imaging reveals nanostructures of signaling complexes

(A) Two-color PALM image of a Jurkat E6.1 cell on a TCR stimulating coverslip. The cell expressed LAT-Dronpa and TCRu03b6-PAmCherry. Bar - 2 u03bcm. (B) Bivariate pair correlation function [PCF or g 1...

Fig. 5

T-cell activation is driven by the dynamic nanoscale organization of signaling clusters

(A) TCR and LAT reside in small nanoclusters within lamellae. As a lamella touches the stimulatory surface, small signaling clusters composed of nanoclusters form at the point of contact. Phosphorylat...

Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.

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