The EAE was induced in female C57BL/6 mice by the "Salari Institute of Cognitive and Behavioral Disorders" as previously described (31) . Following a ketamine/xylazine (Merck, Germany) anesthesia, all animals were immunized according to the previous protocol described by Aghaie T, et al. (17) . After 48 h of the immunization, all animals have received intraperitoneal (IP) pertussis toxin (Sigma-Aldrich, P7208).
Clinical signs and bodyweight of mice were assessed daily by a blind observer. Symptoms were scored according to Table 1. 25 days after EAE induction, the mice were anesthetized and spleens were removed and single-cell suspension of splenocytes were obtained. In brief, the spleen was perfused and the cells were passed through the cell strainers of 70 and 40µm. The cells were centrifuged at 300g for 10min, and the supernatant was discarded. To lysis the red blood cells, the splenocytes were incubated with 10mL ACK 1X (ammonium-chloride-potassium) lysing buffer for 5min at 4°C. The mixture was centrifuged at 300g for 5min and the supernatant was removed. After washing once with RPMI-1640 medium, the reminding cell suspensions were the spleen mononuclear cells. Then, all animals were sacri ced according to the ethics protocol and the spinal cord was removed for further histopathological studies.
Clinical signs and bodyweight of mice were assessed daily by a blind observer. Symptoms were scored according to Table 1. 25 days after EAE induction, the mice were anesthetized and spleens were removed and single-cell suspension of splenocytes were obtained. In brief, the spleen was perfused and the cells were passed through the cell strainers of 70 and 40µm. The cells were centrifuged at 300g for 10min, and the supernatant was discarded. To lysis the red blood cells, the splenocytes were incubated with 10mL ACK 1X (ammonium-chloride-potassium) lysing buffer for 5min at 4°C. The mixture was centrifuged at 300g for 5min and the supernatant was removed. After washing once with RPMI-1640 medium, the reminding cell suspensions were the spleen mononuclear cells. Then, all animals were sacri ced according to the ethics protocol and the spinal cord was removed for further histopathological studies.