Experimental autoimmune encephalomyelitis impairs the immunomodulatory effects of mesenchymal stem cells on splenocyte responses.

backgroundThe effect of disease conditions on the differentiation capability of mesenchymal stem cells (MSCs) has been extensively studied. However, the mechanism by which disease-related alterations in MSCs affect the immune response has not been reported. METHODS AND

resultsMSCs were isolated from adipose tissue of healthy and EAE-induced mice and cultured with or without preconditioning usinglipopolysaccharide (LPS, 10 ng/mL). The immunomodulatory effects of MSC-conditioned media (CM) on the recall responses of splenocytes were evaluated. Cytokine levels, including IL-10, TGF-β, IL-17a, and nitric oxide (NO), were measured in the culture supernatants using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of transcription factors Foxp3, Gata3, Rorc, IFN-γ, and Tbx21 in the splenocytes were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). MSCs from healthy mice produced higher levels of IL-10 than those from EAE mice, with no significant difference in TGF-β production. Splenocytes treated with MSC-CM from healthy mice had higher Foxp3 and Gata3 expression, along with increased IL-10 and NO production, than those treated with EAE MSC-CM. In contrast, splenocytes treated with EAE MSC-CM showed higher Rorc expression and IL-17a levels. LPS preconditioning of EAE MSCs partially restored their immunomodulatory capacity, increased IL-10 levels, and decreased IL-17a levels. However, IFN-γ and Tbx21 expression levels remained similar in splenocytes treated with MSC-CM from both healthy and EAE mice.

conclusionMSCs from EAE mice showed a reduced capacity to upregulate Foxp3 mRNA and modulate immune responses in restimulated splenocytes compared to healthy MSCs. LPS preconditioning partially restored immunomodulatory properties. CLINICAL TRIAL NUMBER: Not applicable.