However, there was a delay in EAE clinical score until day 12 in mice gavaged with MMF. pathway responses, FHF3 thereby exerting neuroprotective effect by Nrf-2 mediated protection in MS tissues . In EAE, DMF ameliorated the clinical course in myelin oligodendrocyte glycoprotein (MOG)-induced EAE in C57BL/6 mice. In addition, DMF suppressed Th1 and Th17 cell differentiation, as well as expression of pro-inflammatory cytokines IFN-, TNF-, and IL-17 [25,26]. It also promoted Th2 cells that produce IL-4, IL-5, and IL-10 . Additionally, vitamin D3 impaired dendritic cells (DCs) maturation which leads to reducing antigen presentation for encephalitogenic CD4+ T cells , and subsequently protecting the mice from developing EAE . Recently, we explained that MMF augments main human CD56+ NK cell lysis SBC-115076 of K562 and SBC-115076 RAJI tumor cells . However, the effect of MMF in MS patients and the mouse model EAE has not been clearly defined. SBC-115076 The present work compares the effects of vitamin D3 and MMF in mice with EAE. 2. Results The protocol for the study design is usually shown in Physique 1. EAE was induced in SJL mice, and as a control normal SBC-115076 mice were used. The first group of mice was left untreated, while the second was treated with vitamin D3, and the third group was fed with MMF, as shown in Physique 1. Open in a separate windows Physique 1 An overview of the study design. Mice were inoculated at day 0 with antigen and pertussis toxin (PTX). They were either left untreated or were treated with vitamin D3 at the indicated time points (blue arrows), or were fed with monomethyl fumarate (MMF) every day throughout the study. After 7 days, bone marrow was harvested to generate immature dendritic cells (iDCs) and mature dendritic cells (mDCs). At day 15 the spleens were isolated to generate natural killer (NK) cells which were used in the cytotoxicity assay. 2.1. Vitamin D3 or MMF Reduces the EAE Clinical Score First we sought to demonstrate if injecting the mice with vitamin D3 or feeding them with MMF might reduce the incidence of EAE. During the 50 days of measuring the EAE clinical score, it was observed that injecting vitamin D3 significantly reduced the EAE clinical score in these mice (< 0.01, Physique 2). However, the best reduction in the EAE clinical score was observed in mice fed with MMF (< 0.0001 as compared to EAE mice that were left untreated, Figure 2A). Comparable outcome was observed when the data were evaluated by area under curve analysis (Physique 2B). Open in a separate window Open in a separate window Physique 2 Comparison of the experimental autoimmune encephalomyelitis (EAE) clinical score among untreated mice (reddish collection), mice treated with vitamin D3 (green collection) or those fed MMF (pink collection) for 50 days. The significant values were calculated using one of the ways ANOVA followed by Sidaks multiple comparison test during the entire period of the experiments (A). Results were also evaluated by area under curve analysis (B). Data were collected from 10 mice in each group at any time point. 2.2. Vitamin D3 Induces NK Cell Lysis of Dendritic Cells In this series of experiments, we sought to confirm the effect of injecting vitamin D3 around the development of EAE in SJL mice and correlated this effect with the ability of the drug to activate NK cells. As shown in Physique 3A, the.