Hobit identifies tissue-resident memory T cell precursors that are regulated by Eomes

Tissue-resident memory CD8⁺ T cells (T_RM) constitute a noncirculating memory T cell subset that provides early protection against reinfection. However, how T_RM arise from antigen-triggered T cells has remained unclear. Exploiting the T_RM-restricted expression of Hobit, we used T_RM reporter/deleter mice to study T_RM differentiation. We found that Hobit was up-regulated in a subset of LCMV-specific CD8⁺ T cells located within peripheral tissues during the effector phase of the immune response. These Hobit⁺ effector T cells were identified as T_RM precursors, given that their depletion substantially decreased T_RM development but not the formation of circulating memory T cells. Adoptive transfer experiments of Hobit⁺ effector T cells corroborated their biased contribution to the T_RM lineage. Transcriptional profiling of Hobit⁺ effector T cells underlined the early establishment of T_RM properties including down-regulation of tissue exit receptors and up-regulation of T_RM-associated molecules. We identified Eomes as a key factor instructing the early bifurcation of circulating and resident lineages. These findings establish that commitment of T_RM occurs early in antigen-driven T cell differentiation and reveal the molecular mechanisms underlying this differentiation pathway.