Human Th2 but not Th9 cells release IL-31 in a STAT6/NF-κB-dependent way

IL-31, a member of the IL-6 protein family, is one of the latest additions to the list of T cell-derived cytokines. Th2 cells are regarded as a main source of IL-31, which is produced in response to stimulation by IL-4. Because the development of Th9 cells also requires IL-4 as a polarizing cytokine, the current study investigates IL-31 production in human Th9 cells compared with Th2 cells. We found that, although Th9 cells were able to release IL-31 during the first weeks of in vitro polarization, no IL-31 was detected in Th9 cultures after a final restimulation in the absence of polarizing cytokines. We further show that TGF-β, which is required to obtain Th9 cells in vitro, potently inhibits the release of IL-31 from Th2 cells, whereas IL-33, a cytokine associated with Th2-mediated inflammation, synergizes with IL-4 in inducing IL-31 secretion. To analyze the molecular mechanisms underlying the induction of IL-31, EMSAs, reporter gene assays, and small interfering RNA-based silencing experiments were carried out. We show that STAT6 and NF-κB are central players in mediating IL-31 expression induced by IL-4/IL-33. In addition, we identified a novel NF-κB-binding element within the Il31 promoter that mediates the enhancing effects of IL-33 on IL-4/STAT6-induced IL-31 expression in human Th2 cells. Taken together, this study shows that IL-4 is essential for the production of IL-31, whereas TGF-β significantly suppresses IL-31 expression at the mRNA and protein levels. As a consequence, in vitro polarized Th2 cells, but not Th9 cells, are able to release IL-31.