Fig. 4

Reduction in inflammation and modulation of Treg and Th17 cells by the transplanted cells. a, b Histological analysis of paraffin sections stained with H&E and inflammation scores of the brain and spinal cord, respectively. Cellular infiltrates reduced significantly in the CNS in EAE animals transplanted with MSCs and NSCs. Symbols, **, &, §, and ¥ indicate significant difference at p ≤ 0.01 between all experimental conditions: healthy control, EAE, EAE + MSCs, and EAE + NSCs. All scale bars represent 50 μm (magnification: 40 ×). c Immunohistochemical staining of paraffin sections of the brain and spinal cord with cell-specific antibodies, CD45, CD68, and CD3E, representing leukocytes, macrophages, and T cells, respectively. The inserts (on the right) were magnified and overexposed to enhance contrast. All scale bars represent 50 μm (magnification: 40 ×). d, e Quantification of fluorescent intensity of brain and spinal cord sections shown in c, respectively (**p ≤ 0.01). f–h Transcription of inflammation markers, Cd3e, Il-17a, Il-2, Il-1β, Il-6, Tnfα, and Ifnγ, in the brain, spinal cord, and spleen, using qRT-PCR. Fold gene expression was normalized to Gapdh and β-Actin and error bars represent the SEM of triplicate measures (**p ≤ 0.01). i Graphical representation of Treg (CD4 + CD25 + FOXP3 +) and Th17 (CD4 + IL-17A +) cells in the blood, spleen, brain, and spinal cord as determined by flow cytometry (**p ≤ 0.01). Treg cell numbers went down, but Th17 went up in the blood and spleen, and both of these cells were significantly increased in the CNS of EAE animals. Transplanted cells significantly increased Treg but significantly reduced Th17 cells in the blood and spleen, and both of these cells were significantly reduced in the CNS of EAE animals. j Splenocytes isolated from control and EAE mice were cultured and treated with MOG_35–55. After 72 h following MOG activation, the cultures were treated with the MSCs and NSCs, and splenocyte proliferation was determined using BrdU assay