IFN-beta and sIL-1Ra play crucial roles in the regulation of innate immunity and inflammation. IFN-beta, which is widely used to improve the course of relapsing, remitting multiple sclerosis, induces the production of sIL-1Ra in human monocytes through mechanisms that remain largely unknown. In this study, we identified PI3Kdelta and MEK2 as key elements that control sIL-1Ra production in isolated human monocytes activated by IFN-beta. Blockade of MEK2, but not of MEK1, by inhibitors and siRNA prevented IFN-beta-induced PI3Kdelta recruitment to the membrane, Akt phosphorylation, and sIL-1Ra production, suggesting that MEK2 acted upstream of PI3Kdelta. Furthermore, ERK1/2, the only identified substrates of MEK1/2 to date, are dispensable for sIL-1Ra production in response to IFN-beta stimulation. Upon IFN-beta activation, MEK2 and PI3Kdelta are translocated to monocyte membranes. These data suggest that MEK1 and MEK2 display different, nonredundant functions in IFN-beta signaling. That neither MEK1 nor ERK1/2 play a part in this mechanism is also an unexpected finding that gives rise to a better understanding of the MAPK signaling network. Together, these findings demonstrate that IFN-beta triggers an atypical MEK2/PI3Kdelta signaling cascade to regulate sIL-1Ra expression in monocytes. The premise that MEK1 and MEK2 play a part in the induction of the proinflammatory cytokine, IL-1beta in human monocytes provides a rationale for an alternative, IFN-beta-mediated pathway to induce/enhance sIL-1Ra production and thus, to dampen inflammation.