The development of T helper (T_H)17 and regulatory T (T_reg) cells is reciprocally regulated by cytokines. Transforming growth factor (TGF)-β alone induces FoxP3⁺ T_reg cells, but together with IL-6 or IL-21 induces T_H17 cells. Here we demonstrate that IL-9 is a key molecule that affects differentiation of T_H17 cells and T_reg function. IL-9 predominantly produced by T_H17 cells, synergizes with TGF-β1 to differentiate naïve CD4⁺ T cells into T_H17 cells, while IL-9 secretion by T_H17 cells is regulated by IL-23. Interestingly, IL-9 enhances the suppressive functions of FoxP3⁺ CD4⁺ T_reg cells in vitro, and absence of IL-9 signaling weakens the suppressive activity of nT_regs in vivo, leading to an increase in effector cells and worsening of experimental autoimmune encephalomyelitis. The mechanism of IL-9 effects on T_H17 and T_regs is through activation of STAT3 and STAT5 signaling. Our findings highlight a role of IL-9 as a regulator of pathogenic versus protective mechanisms of immune responses.