Background: Multiple Sclerosis (MS) is an autoimmune disease of the central nervous system. Autoreactive T cells including cells with a Th17 phenotype are critical players in MS pathogenesis. In this study, we investigated the effects of VitA/D on miRNAs expression involved in Th17 development neuroinflammation using (EAE). Methods: EAE was induced in C57BL/6 mice and received IP injections of vitamins A, D or their combination starting one day before the immunization and continued every other day for 30 days. Animals were scored for 30 days. Percentages of Th17 cells were measured in splenocytes following in vitro re-stimulation with MOG using intracellular staining and flow cytometry. Expression of miR-98-5p and Let-7a-5p, two miRNAs that are known to target Ror-t and Ror-t was measured in MOG-stimulated splenocytes as well as in spinal cord tissues using real-time RT-PCR. Results: Treated mice showed decreased frequency of Th17 cells in their spleens following in vitro re-stimulation with antigen, also lower expression of IL17 and Ror-t in their in CNS and splenocytes. Vitamin A and vitamin D-treated splenocytes showed significant upregulation of miR-98-5p in 24 hour and 48 hours time-points and Let-7a-5p expression was induced at 48-hour post-treatment in MOG-treated cells, which showed a strong negative correlation with splenocyte Ror-t levels. Conclusion: Our data suggest that treatment with vitamins A and D can decreased differentiation of Th17 phenotype. This is likely due to upregulation of Ror-t-targeting miRNAs, miR-98-5p and Let-7a-5p following treatment. These findings point to a potential protective role for miRNAs in the context of autoimmune neuroinflammation.