Functional role of T-cell receptor nanoclusters in signal initiation and antigen discrimination

Pageon, Sophie V.; Tabarin, Thibault; Yamamoto, Yui; Ma, Yuanqing; Bridgeman, John S.; Cohnen, André; Benzing, Carola; Gao, Yijun; Crowther, Michael D.; Tungatt, Katie; Dolton, Garry; Sewell, Andrew K.; Price, David A.; Acuto, Oreste; Parton, Robert G.; Gooding, J. Justin; Rossy, Jérémie; Rossjohn, Jamie; Gaus, Katharina

doi:10.1073/pnas.1607436113

Cited by 207 publications

(258 citation statements)

References 71 publications

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“…Therefore, we hypothesize NK signal integration occurs by the relative sizedependent colocalization of receptors at the NK IS, and this is the explanation for the functional differences observed for elongated ligands. Considering the differences observed at a nanometer-scale, our results further support recent evidence that immune cell receptors show a nanometer-scale organization, with functional associated implications [19,22,50,[52][53][54][55]. Interestingly, there is mounting evidence that the molecular mechanisms behind immune inhibition are in part spatially restricted and common to various immune cells such as T, B and NK cells [56][57][58][59].…”

supporting

confidence: 86%

Nanoscale colocalization of NK cell activating and inhibitory receptors controls signal integration

Tomaz

Pereira

Guerra

et al. 2018

Preprint

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NK cell responses depend on the balance of signals from inhibitory and activating receptors. However, how the integration of antagonistic signals occurs upon NK cell-target cell interaction is not fully understood. Here, we provide evidence that NK cell inhibition via the inhibitory receptor Ly49A is dependent on its relative colocalization at nanometer-scale with the activating receptor NKG2D upon immune synapse (IS) formation. NKG2D and Ly49A signal integration and colocalization was studied using NKG2D-GFP and Ly49A-RFP-expressing primary NK cells, forming ISs with NIH3T3 target cells, with or without expression of single chain trimer (SCT) H2-D d and an extended form of SCT H2-D d -CD4 MHC-I molecules. Nanoscale colocalization was assessed by Förster resonance energy transfer (FRET) between NKG2D-GFP and Ly49A-RFP and measured for each synapse. In the presence of their respective cognate ligands, NKG2D and Ly49A colocalize at a nanometer scale leading to NK cell inhibition. However, increasing the size of the Ly49A ligand reduced the nanoscale colocalization with NKG2D consequently impairing Ly49A-mediated inhibition. Thus, our data shows NK cell signal integration is critically dependent on the dimensions of NK cell ligand-receptor pairs by affecting their relative nanometer-scale colocalization at the IS. Together, our results suggest the balance of NK cell signals, and NK cell responses, are determined by the relative nanoscale colocalization of activating and inhibitory receptors in the immune synapse. NK cell signal integration | FRET nanoscale localization | NKG2D | Ly49ACorrespondence:david.tomaz@kcl.ac.uk, r.henriques@ucl.ac.uk, k.gould@imperial.ac.uk Introduction NK cell activation is dependent on a different array of germline-encoded receptors capable of triggering effector responses against infection [1][2][3] and cellular transformation [4,5], and yet maintaining tolerance towards healthy cells and tissues [6,7]. NK cell functionality is widely characterized by a balance between activating and inhibitory receptors [8,9]. However, upon immune synapse formation, how NK cells integrate signals from functionally antagonistic receptors, is not yet fully understood [10]. One major hypothesis to explain immune signal integration is based on the kinetic segregation (K-S) model [11]. This model states that the balance of antagonistic signals is mediated by the equilibrium of phosphorylation (kinases) and dephosphorylation (phosphatases), which depends on the relative colocalization of receptors and their associated signaling molecules upon synapse formation. This hypothesis has been validated using T cells, in the context of TCR triggering [12,13] and, more recently, NK cells [14,15]. The K-S model suggests immune inhibition, upon immune synapse formation, is maintained by the dephosphorylation of tyrosines in stimulatory receptors (e.g ITAMs-associated receptors such as TCR-CD3 or NKG2D-DAP12), due to a nanoscale colocalization with receptors bearing phosphatase activity (e.g. CD45, CD148) or with t...

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supporting

confidence: 86%

Nanoscale colocalization of NK cell activating and inhibitory receptors controls signal integration

Tomaz

Pereira

Guerra

et al. 2018

Preprint

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“…Imaging of these surrogate substrates has revealed the segregation of different immune receptor pairs into concentric regions called central supramolecular activation clusters (cSMACs), peripheral SMACs ( pSMACs) and distal SMACs (dSMACs). This demonstrates that the actin-based spatial segregation of the different receptor signals is key to eliciting the immunological response of the T-cell (Dustin and Groves, 2012;Groves, 2007;Pageon et al, 2016;Tanaka and Sackmann, 2005;Yu and Groves, 2010).…”

Section: Ecm Sensingmentioning

confidence: 88%

How cells respond to environmental cues – insights from bio-functionalized substrates

Ruprecht

Monzo

Ravasio

et al. 2016

Journal of Cell Science

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Biomimetic materials have long been the (he)art of bioengineering. They usually aim at mimicking in vivo conditions to allow in vitro culture, differentiation and expansion of cells. The past decade has witnessed a considerable amount of progress in soft lithography, bioinspired micro-fabrication and biochemistry, allowing the design of sophisticated and physiologically relevant micro-and nanoenvironments. These systems now provide an exquisite toolbox with which we can control a large set of physicochemical environmental parameters that determine cell behavior. Biofunctionalized surfaces have evolved from simple protein-coated solid surfaces or cellular extracts into nano-textured 3D surfaces with controlled rheological and topographical properties. The mechanobiological molecular processes by which cells interact and sense their environment can now be unambiguously understood down to the single-molecule level. This Commentary highlights recent successful examples where bio-functionalized substrates have contributed in raising and answering new questions in the area of extracellular matrix sensing by cells, cell-cell adhesion and cell migration. The use, the availability, the impact and the challenges of such approaches in the field of biology are discussed.

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“…This hypothesis is in accordance with the work of others showing that aggregation of CD3z molecules promotes T cell activation. 42,43 If this hypothesis is true, the stronger activation stimulus could lead to the observed functional differences in T cells expressing Hu19-CD828Z versus Hu19-28Z, such as the differences in cytokine production and AICD. As we learn more about CAR functionality and the importance of hinge domains on CAR function, it is possible that optimal hinges from either CD28 or CD8a could be selected for different indications or target antigens.…”

Section: E) T Cells From Six Different Donors Expressing Eithermentioning

confidence: 99%

Function of Novel Anti-CD19 Chimeric Antigen Receptors with Human Variable Regions Is Affected by Hinge and Transmembrane Domains

et al. 2017

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Anti-CD19 chimeric antigen receptor (CAR) T cells have caused remissions of B cell malignancies, but problems including cytokine-mediated toxicity and short persistence of CAR T cells in vivo might limit the effectiveness of anti-CD19 CAR T cells. Anti-CD19 CARs that have been tested clinically had single-chain variable fragments (scFvs) derived from murine antibodies. We have designed and constructed novel anti-CD19 CARs containing a scFv with fully human variable regions. T cells expressing these CARs specifically recognized CD19 target cells and carried out functions including degranulation, cytokine release, and proliferation. We compared CARs with CD28 costimulatory moieties along with hinge and transmembrane domains from either the human CD28 molecule or the human CD8α molecule. Compared with T cells expressing CARs with CD28 hinge and transmembrane domains, T cells expressing CARs with CD8α hinge and transmembrane domains produced lower levels of cytokines and exhibited lower levels of activation-induced cell death (AICD). Importantly, CARs with hinge and transmembrane regions from either CD8α or CD28 had similar abilities to eliminate established tumors in mice. In anti-CD19 CARs with CD28 costimulatory moieties, lower levels of inflammatory cytokine production and AICD are potential clinical advantages of CD8α hinge and transmembrane domains over CD28 hinge and transmembrane domains.

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Functional role of T-cell receptor nanoclusters in signal initiation and antigen discrimination

Cited by 207 publications

References 71 publications

Nanoscale colocalization of NK cell activating and inhibitory receptors controls signal integration

Nanoscale colocalization of NK cell activating and inhibitory receptors controls signal integration

How cells respond to environmental cues – insights from bio-functionalized substrates

Function of Novel Anti-CD19 Chimeric Antigen Receptors with Human Variable Regions Is Affected by Hinge and Transmembrane Domains

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