Hypoxia acts as an environmental cue for the human tissue-resident memory T cell differentiation program

Tissue-resident memory T cells (T_RM) provide frontline defense against infectious diseases and contribute to antitumor immunity; however, aside from the necessity of TGF-β, knowledge regarding T_RM-inductive cues remains incomplete, particularly for human cells. Oxygen tension is an environmental cue that distinguishes peripheral tissues from the circulation, and here, we demonstrate that differentiation of human CD8+ T cells in the presence of hypoxia and TGF-β1 led to the development of a T_RM phenotype, characterized by a greater than 5-fold increase in CD69+CD103+ cells expressing human T_RM hallmarks and enrichment for endogenous human T_RM gene signatures, including increased adhesion molecule expression and decreased expression of genes involved in recirculation. Hypoxia and TGF-β1 synergized to produce a significantly larger population of T_RM phenotype cells than either condition alone, and comparison of these cells from the individual and combination conditions revealed distinct phenotypic and transcriptional profiles, indicating a programming response to milieu rather than a mere expansion. Our findings identify a likely previously unreported cue for the T_RM differentiation program and can enable facile generation of human T_RM phenotype cells in vitro for basic studies and translational applications such as adoptive cellular therapy.