Nanoscale Ligand Spacing Influences Receptor Triggering in T Cells and NK Cells

Delcassian, Derfogail; Depoil, David; Rudnicka, Dominika; Liu, Mengling; Davis, Daniel M.; Dustin, Michael L.; Dunlop, Iain E.

doi:10.1021/nl403252x

Cited by 116 publications

(124 citation statements)

References 68 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].…”

Section: Discussionsupporting

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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Section: Discussionsupporting

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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“…The mean interparticle spacing is 48 ± 8 nm (Fig. 1C), which is physiologically optimal for antigen-mediated T-cell adhesion and functional responses (14,15). Each particle presents an average of 4 ± 1 DNA hairpins (SI Appendix, Fig.…”

Section: Resultsmentioning

confidence: 99%

DNA-based nanoparticle tension sensors reveal that T-cell receptors transmit defined pN forces to their antigens for enhanced fidelity

Liu

Blanchfield

Pui‐Yan

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

287

347

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T cells are triggered when the T-cell receptor (TCR) encounters its antigenic ligand, the peptide-major histocompatibility complex (pMHC), on the surface of antigen presenting cells (APCs). Because T cells are highly migratory and antigen recognition occurs at an intermembrane junction where the T cell physically contacts the APC, there are long-standing questions of whether T cells transmit defined forces to their TCR complex and whether chemomechanical coupling influences immune function. Here we develop DNA-based gold nanoparticle tension sensors to provide, to our knowledge, the first pN tension maps of individual TCR-pMHC complexes during T-cell activation. We show that naïve T cells harness cytoskeletal coupling to transmit 12–19 pN of force to their TCRs within seconds of ligand binding and preceding initial calcium signaling. CD8 coreceptor binding and lymphocyte-specific kinase signaling are required for antigen-mediated cell spreading and force generation. Lymphocyte function-associated antigen 1 (LFA-1) mediated adhesion modulates TCR-pMHC tension by intensifying its magnitude to values >19 pN and spatially reorganizes the location of TCR forces to the kinapse, the zone located at the trailing edge of migrating T cells, thus demonstrating chemomechanical crosstalk between TCR and LFA-1 receptor signaling. Finally, T cells display a dampened and poorly specific response to antigen agonists when TCR forces are chemically abolished or physically “filtered” to a level below ∼12 pN using mechanically labile DNA tethers. Therefore, we conclude that T cells tune TCR mechanics with pN resolution to create a checkpoint of agonist quality necessary for specific immune response.

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“…Recent studies have further highlighted the importance of nanoscale ligand geometry in immune system responses [ 109 , 110 ]. For example, using the block copolymer micelle lithography (BCML) approach, Delcassian et al [ 109 ] generated CD3 and CD16 antibody patterns to monitor the effect of ligand spacing on the activation levels of T cells and Natural Killer (NK) cells. The activations were assessed in terms of membrane-localized phosphotyrosine for T cells and the size of contact area for the NK cells, and the study showed decreased cell response with increasing ligand spacing in both cell types, with the threshold activation at the ligand spacing being 69 nm and 104 nm for T cells and NK cells, respectively.…”

Section: Biomolecular Cuesmentioning

confidence: 99%

Guided Cellular Responses by Surface Cues for Nanomedicine Applications

Ogaki

Andersen

Foss

2016

Advances in Delivery Science and Technology

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Using surface cues to guide and ultimately control cellular responses is of paramount importance in numerous biomedical applications. Since cells react on feature sizes both on the micro and nanometer scale, these length scales are crucial parameters when designing new materials for applications in medicine e.g. for tissue engineering and drug delivery. Thus, variation of the features at the nanometer length scale is an integral part of nanomedicine research and development. In this chapter the interaction between biological systems and artifi cial materials will be addressed in general with focus on simplifi ed model systems where only one or a few parameters are varied at two-dimensional (2D) surfaces. At fi rst biomolecular adsorption/immobilization on surfaces will be addressed followed by a discussion of approaches to synthesize functionalized surfaces and the infl uence of such surfaces on cellular response. Some of the key parameters, which will be discussed in more detail, are topography, chemistry, and elastic modulus of the substrate. Even though there is a vast amount of published data it will become clear that there is still not a detailed understanding of the infl uence of these parameters on biosystems at the cellular level. Obviously, the degree of complexity increases when several of the surface cues are combined. The challenges in understanding the cellular responses in detail in real systems with the simultaneous variation of multiple parameters leads to the introduction of the fi eld of high throughput screening of biomaterials.

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Nanoscale Ligand Spacing Influences Receptor Triggering in T Cells and NK Cells

Cited by 116 publications

References 68 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

DNA-based nanoparticle tension sensors reveal that T-cell receptors transmit defined pN forces to their antigens for enhanced fidelity

Guided Cellular Responses by Surface Cues for Nanomedicine Applications

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