How drag sharpens a T cell’s view on antigen

Schütz, Gerhard J.; Huppa, Johannes B.

doi:10.1073/pnas.1911389116

Cited by 13 publications

(7 citation statements)

References 19 publications

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“…We also have evidence that CTLs stimulated with anti-CD3 vs. cognate peptide-MHC complexes behave similarly in our functional readouts (see materials and methods in [13]. Physical parameters such as mechanical forces, however, do influence agonist TCR-pMHC interactions occurring during T cell priming processes [25]. While this is something we cannot address in our study, it would provide more information about the function of the cytolytic IS.…”

Section: Discussionmentioning

confidence: 89%

Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus

Estl

Blatt

et al. 2020

IJMS

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Cytotoxic T lymphocytes (CTL) are key players of the adaptive immune system that target tumors and infected cells. A central step to that is the formation of a cell–cell contact zone between the CTL and its target called an immune synapse (IS). Here, we investigate the influence of the initial T cell receptor (TCR) trigger of a cytolytic IS on the distinct steps leading to cytotoxic granule (CG) exocytosis. We stimulated primary CTLs from mouse using lipid bilayers with varying anti-CD3 but constant ICAM concentrations. We fluorescently labeled molecular markers of distinct IS zones such as actin, CD3, granzyme B, and Synaptobrevin2 in CTLs and imaged cytolytic IS formation by total internal reflection fluorescence microscopy (TIRFM). We found that an intermediate anti-CD3 concentration of 10 µg/mL induces the fastest adhesion of CTLs to the bilayers and results in maximal CG fusion efficiency. The latency of actin ring formation, dwell time, and maximum surface area at the IS exhibit different dependencies on the stimulatory anti-CD3 concentrations. The number and surface area of CD3 clusters at the IS seem to show a different dependency to the TCR trigger when compared to their dwell time. Finally, the mode of full CG exocytosis appears to be independent of the TCR trigger.

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Section: Discussionmentioning

confidence: 89%

Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus

Estl

Blatt

et al. 2020

IJMS

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“…Such force ramps not only affect the mean but particularly the distribution of bond lifetimes, an effect which could be exploited by T-cells to improve the specificity and sensitivity of antigen discrimination (4,48).…”

Section: Discussionmentioning

confidence: 99%

Temporal Analysis of T-Cell Receptor-Imposed Forces via Quantitative Single Molecule FRET Measurements

Göhring

Kellner

Schrangl

et al. 2020

Preprint

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Mechanical forces acting on ligand-engaged T-cell receptors (TCRs) have been implicated in T-cell antigen recognition. To measure these molecular forces in real time, we devised a FRET-based force sensor that is equipped with a TCR-reactive single chain antibody fragment and tethered to planar supported lipid bilayers (SLBs) which serve as surrogate antigen presenting cell. When confronting T-cells with sensorfunctionalized gel-phase SLBs, we measured 4.6 pN force-peaks for activated T-cells and 5.5 pN forcepeaks for non-activated scanning T-cells, which vary in the range of seconds with force loads of 1.5 pN per second. When using more physiological fluid-phase SLBs, TCR-imposed forces were found to have a ~3fold reduction for scanning T-cells, yet no longer detectable for activated T-cells. Hence, if they are essential for ligand discrimination and TCR-proximal signaling during initial antigen scanning, TCRexerted pulling forces are already effective below 2pN. Once activated, T-cells appear to create a synaptic environment that is devoid of noticeable TCR-exerted mechanical forces.Göhring J. et al., page 2 of 54

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“…4c ). Such force ramps should not only affect the mean but in particular the distribution of bond lifetimes, a property which could be exploited by T-cells to improve the specificity and sensitivity of antigen discrimination 4 , 51 . Previously, we have reasoned by analytical deduction that loading rates of only 1 pN/s would already suffice to enable peptide discrimination with high specificity and sensitivity 4 .…”

Section: Discussionmentioning

confidence: 99%

Temporal analysis of T-cell receptor-imposed forces via quantitative single molecule FRET measurements

et al. 2021

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Mechanical forces acting on ligand-engaged T-cell receptors (TCRs) have previously been implicated in T-cell antigen recognition, yet their magnitude, spread, and temporal behavior are still poorly defined. We here report a FRET-based sensor equipped either with a TCR-reactive single chain antibody fragment or peptide-loaded MHC, the physiological TCR-ligand. The sensor was tethered to planar glass-supported lipid bilayers (SLBs) and informed most directly on the magnitude and kinetics of TCR-imposed forces at the single molecule level. When confronting T-cells with gel-phase SLBs we observed both prior and upon T-cell activation a single, well-resolvable force-peak of approximately 5 pN and force loading rates on the TCR of 1.5 pN per second. When facing fluid-phase SLBs instead, T-cells still exerted tensile forces yet of threefold reduced magnitude and only prior to but not upon activation.

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How drag sharpens a T cell’s view on antigen

Cited by 13 publications

References 19 publications

Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus

Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus

Temporal Analysis of T-Cell Receptor-Imposed Forces via Quantitative Single Molecule FRET Measurements

Temporal analysis of T-cell receptor-imposed forces via quantitative single molecule FRET measurements

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