T Cell Activation is Determined by the Number of Presented Antigens

Deeg, Janosch; Axmann, Markus; Matic, Jovana; Liapis, Anastasia; Depoil, David; Afrose, Jehan; Curado, Silvia; Dustin, Michael L.; Spatz, Joachim P.

doi:10.1021/nl403266t

Cited by 121 publications

(137 citation statements)

References 54 publications

(106 reference statements)

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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.

278

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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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.

278

347

View full text Add to dashboard Cite

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“…Human integrin αvβ3 was incubated for 24 h in the absence of competition, while fluorescence measurements were conducted in the presence of c(-RGDfK-) 100 μM. All experiments were performed at 37 °C, using a microscopy system previously described [73]. Details concerning incubation and washing conditions, as well as measurement conditions and settings, are presented in Supporting Information.…”

Section: In Vitro αVβ3 Integrin Binding Assaysmentioning

confidence: 99%

Focal adhesion stabilization by enhanced integrin-cRGD binding affinity

et al. 2017

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Abstract:In this study we investigate the impact of ligand presentation by various molecular spacers on integrin-based focal adhesion formation. Gold nanoparticles (AuNPs) arranged in hexagonal patterns were biofunctionalized with the same ligand head group, cyclic Arg-Gly-Asp [c(-RGDfX-)], but with different molecular spacers, each of which couples the head group to the gold. Aminohexanoic acid, polyethylene glycol (PEG) and polyproline spacers were used to vary the distance between the binding motif and the substrate, and thus the presentation of integrin binding on anchoring points. Adherent cells plated on nanopatterned surfaces with polyproline spacers for peptide immobilization could tolerate larger ligand spacing (162 nm) for focal adhesion formation, in comparison to cells on surfaces with PEG (110 nm) or aminohexanoic acid (62 nm) spacers. Due to the rigidity of the polyproline spacer, enhanced access to the ligand-binding site upon integrin-cRGD complex formation increases the probability of rebinding and decreases unbinding, as measured by fluorescence recovery after photobleaching (FRAP) analysis, compared to the analogues with aminohexanoic acid or PEGcontaining spacers. These findings indicate that focal adhesion formation may not only be stabilized upon tight integrin clustering, but also by tuning the efficiency of the exposure of the cRGD-based ligand to the integrin extracellular domains. Our studies clearly highlight the importance of ligand spatial presentation for regulating adhesion-dependent cell behavior, and provide a sound approach for studying cell signaling processes on nanometer-scale, engineered bioactive surfaces under chemical stimuli of varying intensities.

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“…Nanofabricated surfaces have previously been used to study the effect of ligand spacing on TCR triggering 9, 10, 11 . Those studies used extended arrays (meaning much larger than the cell) of gold nanoparticle (NP)-bound TCR ligands surrounded by a non-adhesive background 9, 10 .…”

mentioning

confidence: 99%

“…Those studies used extended arrays (meaning much larger than the cell) of gold nanoparticle (NP)-bound TCR ligands surrounded by a non-adhesive background 9, 10 . A graded increase in TCR triggering with decreasing inter-ligand spacing between 200 nm and 40 nm was observed, but there was no evidence of a spacing threshold 10, 11 .…”

mentioning

confidence: 99%

Full control of ligand positioning reveals spatial thresholds for T cell receptor triggering

et al. 2018

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Elucidating the rules for receptor triggering in cell-cell and cell-matrix contacts requires precise control of ligand positioning in three dimensions. Here, we use the T cell receptor (TCR) as a model and subject T cells to different geometric arrangements of ligands, using a nanofabricated single-molecule array platform. This is comprised of monovalent TCR ligands anchored to lithographically patterned nanoparticle clusters surrounded by mobile adhesion molecules on a supported lipid bilayer (SLB). The TCR ligand could be co-planar with the SLB (2D), excluding the CD45 transmembrane tyrosine phosphatase, or elevated by 10 nm on solid nanopedestals (3D), allowing closer access of CD45 to engaged TCR. The two configurations resulted in different T cell responses, depending on the lateral spacing between the ligands. These results identify the important contributions of lateral and axial components of ligand positioning and create a more complete foundation for receptor engineering for immunotherapy.

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T Cell Activation is Determined by the Number of Presented Antigens

Cited by 121 publications

References 54 publications

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

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

Focal adhesion stabilization by enhanced integrin-cRGD binding affinity

Full control of ligand positioning reveals spatial thresholds for T cell receptor triggering

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