IFNγ and proinflammatory cytokines cooperatively license astrocytes to support antigen-dependent CD4+ T cell restimulation.

We report that type 2 interferon with proinflammatory cytokines induces astrocytes to express multiple components required for attraction of and antigen presentation to CD4+ T cells via MHC-II molecules. Re-analysis of astrocyte-enriched single-cell sequencing from bacterial mimetic-injected mice revealed a transcriptionally distinct interferon-responsive astrocyte population enriched for antigen-presentation-associated genes 24 hours after peripheral immune challenge. In vivo, we identify MHC-II+GFAP+ astrocytes localized to the surface of the brain across multiple mouse models of neuroinflammation and in human neurological disease tissue. In a pure in vitro culture system, astrocytes stimulated with IFNγ and proinflammatory cytokines express MHC-II, costimulatory molecules and lymphocyte-recruiting chemokines, and support rapid antigen-dependent expansion of previously activated CD4+ T cells. Using human iPSC-derived CNS cultures, we additionally show induction of antigen-presentation-associated machinery in human astrocytes following inflammatory cytokine stimulation. Rather than inferring function solely from marker expression, we assessed astrocyte-supported T cell restimulation in vitro and separately characterized the emergence of antigen-presentation-associated astrocytes in vivo at neuroimmune interfaces during inflammation in mice and humans. Together, these findings support a conserved role for cytokine-stimulated astrocytes in expressing antigen-presentation-associated machinery and, in vitro, supporting antigen-dependent restimulation/expansion of CD4+ T cells under inflammatory conditions.