Interleukin (IL)-17 is a key inflammatory cytokine in autoimmune disease and may play an important role in the development of experimental autoimmune encephalomyelitis (EAE). In this study, we observed decreased IL-17 and increased IL-27 in EAE rats treated with bone marrow stromal cells (BMSCs). Neutralization of IL-27 resulted in recovery of the BMSCs effect. Adoptive transfer induction of EAE was poor by BMSC-stimulated MNCs, but could be induced by MNCs stimulated by BMSCs under blockade of IL-27 signaling. These results demonstrate that BMSCs may suppress the development of EAE, possibly via secretion of IL-27, which can inhibit IL-17 production or Th17 cell generation.