Ligand-induced segregation from large cell-surface phosphatases is a critical step in γδ TCR triggering

Gamma/delta (γδ) T cells are unconventional lymphocytes that recognize diverse ligands via somatically recombined T cell antigen receptors (γδ TCRs). The molecular mechanism by which ligand recognition initiates γδ TCR signaling, a process known as TCR triggering, remains elusive. Unlike αβ TCRs, γδ TCRs are not mechanosensitive and do not require co-receptors or typical binding-induced conformational changes for triggering. Here, we show that γδ TCR triggering by nonclassical MHC class Ib antigens, a major class of ligands recognized by γδ T cells, requires steric segregation of the large cell-surface phosphatases CD45 and CD148 from engaged TCRs at synaptic close-contact zones. Increasing access of these inhibitory phosphatases to sites of TCR engagement, by elongating MHC class Ib ligands or truncating CD45/148 ectodomains, abrogates TCR triggering and T cell activation. Our results identify a critical step in γδ TCR triggering and provide insight into the core triggering mechanism of endogenous and synthetic tyrosine-phosphorylated immunoreceptors. [Display omitted] •γδ TCR triggering relies on Src kinase-mediated phosphorylation•Size-based segregation of engaged γδ TCRs from large phosphatases is necessary for triggering•γδ TCR triggering conforms to the kinetic-segregation model Li et al. use super-resolution optical imaging and electron microscopy along with size-based manipulations of CD45 distribution at the immunological synapse to show that γδ TCR triggering requires segregation of large cell-surface phosphatases from engaged TCRs at synaptic contacts.