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

Janett Göhring, Florian Kellner, Lukas Schrangl, René Platzer, Enrico Klotzsch, Hannes Stockinger, Johannes B. Huppa () and Gerhard J. Schütz ()
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Janett Göhring: Medical University of Vienna
Florian Kellner: Medical University of Vienna
Lukas Schrangl: TU Wien
René Platzer: Medical University of Vienna
Enrico Klotzsch: TU Wien
Hannes Stockinger: Medical University of Vienna
Johannes B. Huppa: Medical University of Vienna
Gerhard J. Schütz: TU Wien

Nature Communications, 2021, vol. 12, issue 1, 1-14

Abstract: Abstract 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.

Date: 2021
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DOI: 10.1038/s41467-021-22775-z

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