Force-induced site-specific enzymatic cleavage probes reveal that serial mechanical engagement boosts T cell activation

Abstract

The surface of T cells is studded with T cell receptors (TCRs) that are used to scan target cells to identify peptide-major histo-compatibility complexes (pMHCs) signatures of viral infection or cancerous mutation. It is now established that the TCR-pMHC complex is highly transient and experiences mechanical forces that augment the fidelity of T cell activation. An important question in this area pertains to the role of force duration in immune activation. Herein, we report the development of force probes that autonomously terminate tension within a time window following mechanical triggering. Force-induced site-specific enzymatic cleavage (FUSE) probes tune tension duration by controlling the rate of a force-triggered endonuclease hydrolysis reaction. This new capability provides a method to study how accumulated force duration contributes to T cell activation. We screened DNA sequences and identified FUSE probes that disrupt mechanical interactions with F >7.1 piconewtons (pN) between TCRs and pMHCs. Force lifetimes ({tau}F) are tunable from tens of min down to 1.9 min. T cells challenged with FUSE probes presenting cognate antigens with {tau}F of 1.9 min demonstrated dampened markers of early activation, thus demonstrating that repeated mechanical sampling boosts TCR activation. Repeated mechanical sampling F >7.1 pN was found to be particularly critical at lower pMHC antigen densities, wherein the T cell activation declined by 23% with {tau}F of 1.9 min. FUSE probes with F >17.0 pN response showed weaker influence on T cell triggering further showing that TCR-pMHC with F >17.0 pN are less frequent compared to F >7.1 pN. Taken together, FUSE probes allow a new strategy to investigate the role of force dynamics in mechanotransduction broadly and specifically suggest a model of serial mechanical engagement in antigen recognition.