Immune System Of Multiple Sclerosis Patients Research Articles

The immunomodulatory effects of all-trans retinoic acid and docosahexaenoic acid combination treatment on the expression of IL-2, IL-4, T-bet, and GATA3 genes in PBMCs of multiple sclerosis patients

ABSTRACT Objectives Multiple sclerosis (MS) is a potentially disabling autoimmune disease of the central nervous system. Neither the pathogenesis nor the effectiveness of treatment of MS has been fully understood. This in vitro trial evaluated the beneficial immunomodulatory effects of single and combined treatments of all-trans retinoic acid (ATRA) and docosahexaenoic acid (DHA) on the peripheral blood mononuclear cells (PBMCs) of relapsing-remitting MS (RRMS) patients who were receiving interferon beta (IFN-β). Methods The PBMCs of 15 RRMS patients were isolated, cultured, and treated with single and combined treatments of ATRA and DHA. The expressions of IL-2, IL-4, T-bet, and GATA3 genes were evaluated using real-time PCR. Results The results showed that a single treatment of ATRA could significantly suppress the gene expression of the pro-inflammatory cytokine, IL-2 (P < 0.05), and related transcription factor, T-bet (P < 0.001). The gene expression level of the anti-inflammatory cytokine, IL-4, and its transcription factor, GATA3, were not significantly changed. The expression of IL-2 and T-bet genes was significantly decreased in combination treatments of ATRA and DHA (P < 0.001). Significant suppression of IL-2 and T-bet (P < 0.001) was observed in ATRA and DHA combination therapy with half doses of their single treatment, which suggested a synergistic effect of these components. Discussion Co-administration of vitamin A and DHA, an omega-3 fatty acid derivative, may exert a synergistic effect in modulating the immune system in MS patients; however, more studies are needed to evaluate the exact effects and mechanism of their actions on the immune cells.

Antigen B modulates anti-inflammatory cytokines in the EAE model of multiple sclerosis.

Multiple sclerosis (MS) is characterized by the destruction of the blood-brain barrier, loss of myelin sheath, and contribution of inflammatory interleukins such as TNF-alpha, interleukin-17, and interleukin-6. The current study investigated the effect of antigen B of hydatid cyst fluid on the reduction of anti-inflammatory cytokines and nerve conduction velocity in rats with experimental autoimmune encephalomyelitis (EAE)-induced MS. After isolation of antigen B from sterile cyst fluid, the rats were randomly divided into four groups: saline, EAE, EAE + teriflunomide (EAE + TF), and EAE + antigen B (EAE + AngB). The EAE model was induced using cow spinal cord homogenization, in combination with Freund's complete adjuvant. The serum concentration of cytokines including IL-1B and IL-17, IL-10, IL-6, and TNF-X was measured by the ELISA method, and real-time PCR was performed to study gene expression. Electrophysiological, behavioral, and neuropathological tests were also conducted. Nerve conduction velocity and IL-10 concentration were increased in the antigen B group. The results of this study showed that antigen B reduced the inflammatory component of the EAE MS animal model by modulating the immune system compared to teriflunomide, which eventually led to a reduction in symptoms at the behavioral and electrophysiological level. It seems that antigen B plays a critical role in regulating immunity and it can be used as a possible therapeutic agent to modulate the immune system in MS patients. It might be rational to consider hydatid cyst fluid antigen as a modifier in MS.

Dimethyl fumarate treatment in multiple sclerosis: Recent advances in clinical and immunological studies.

Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system (CNS) in which demyelination and neurodegeneration occurs. The immune system of MS patients is characterized by a dysregulation in the balance between pro- and anti-inflammatory immune cells, whereby both the innate and adaptive immune system are involved. Dimethyl fumarate (DMF) was licensed in 2013 as an oral first-line therapy for relapsing-remitting (RR)MS patients. It has a strong efficacy with neuroprotective and immunomodulatory effects and a favourable benefit-risk profile. However, the effects of DMF on the immune system of MS patients were not clear before entering the market. During the last years, numerous in vitro and ex vivo studies have clarified the working mechanism of DMF in MS. Here, we discuss the pharmacokinetics of DMF and its effect on molecular immune-related pathways, which is further linked to the clinical and immunological effects of DMF treatment. The efficacy and safety of DMF treatment for RRMS is discussed as reported from clinical trials. Further, the immunological effects of DMF treatment in RRMS patients are addressed in more detail, including the distribution and function of immune cells. Taken together, evidence from recent studies points to a multifactorial working mechanism of DMF treatment in MS which leads to a restored immune balance favouring a more tolerogenic or anti-inflammatory immune profile.

Dimethyl fumarate induces a persistent change in the composition of the innate and adaptive immune system in multiple sclerosis patients

The effects of dimethyl fumarate (DMF) on the immune system in multiple sclerosis (MS) are not completely elucidated. In this study, an extensive immunophenotypic analysis of innate and adaptive immune cells of DMF-treated MS patients was performed. Peripheral blood immune cell phenotypes were determined using flow cytometry in a follow-up study of 12 MS patients before, after 3 and 12 months of DMF treatment and a cross-sectional study of 25 untreated and 64 DMF-treated MS patients. Direct effects of DMF on B cells were analyzed in vitro. After 12 months of DMF treatment, percentages of monocytes, natural killer cells, naive T and B cells and transitional B cells increased. Percentages of (effector) memory T cells, (non) class-switched memory B cells and double negative B cells decreased together with CD4+ T cells expressing interferon-γ (IFN-γ), granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-17 (IL-17). DMF treatment was fully effective as of 6 months and directly induced apoptosis and decreased expression of costimulatory CD40, antigen presentation molecule MHCII and B cell activating factor receptor (BAFFR) on B cells. DMF induced a persistent change of the immune system of MS patients, directly induced apoptosis and reduced expression of functional markers on B cells.

Differential Effects of Natalizumab and Fingolimod on the Innate Immune System of MS Patients (P04.125)

Objective: To investigate effects of natalizumab (NA) and fingolimod (FTY) on the innate immune system (IIS) in Multiple Sclerosis (MS) patients. Background Dendritic cells (DC) and monocytes (MO) are professional antigen-presenting-cells (APC) orchestrating the IIS and can have profound effects on T- and B-cell effector-function. SlanDC as major human blood-DC population are potent APC and cytokine-producing effector cells taking part in pro-inflammatory processes in MS. As FTY and NA are primarily drugs interfering with lymphocyte redistribution between lymphoid organs vs. blood (FTY) resp. lymphocyte migration throughout the blood-brain-barrier (NA), little is known about effects on the IIS. Design/Methods: SlanDC and MO of healthy controls, untreated MS-patients and MS-patients treated with NA or FTY were analyzed regarding frequency and activation-status. In addition, they were isolated by immunomagnetic-cell-sorting. After stimulation with TLR-ligands, they were analyzed with and without FTY720, FTY720-phosphate or S1P regarding their cytokine-release, T-cell-programming and –priming. Results: Natalizumab-treatment did not change frequency of slanDC, MO and DC1/2 in blood, but lead to reduced monocyte frequency in CSF. Natalizumab could be demonstrated on the surface of all IIS cells. During NA-treatment, no differences in cytokine-release like TNF-a, IL10, IL6 or IL12 and activation-phenotype could be described for slanDC and MO compared to healthy- and untreated-controls. In contrast, fingolimod-treatment lead to an increase of slanDC and MO, whereas DC1/DC2 frequencies were stable. In FTY-treated-patients, pro-inflammatory cytokine-secretion of TNF-a and IL12 was inhibited in slanDC, but not modulated in MO. By in vitro experiments using sorted slanDC and MO, S1P increased and FTY decreased pro-inflammatory cytokine-profiles. FTY impaired the induction of IFN-gamma-producing Th1-cells after LPS-stimulation. Conclusions: Natalizumab only lead to a frequency reduction of cells of the IIS in CSF. FTY did not only modulate T- and B-lymphocyte trafficking, but also effect APC frequency and function. Our results suggest an impact of FTY on T-cell-programming and cytokine-secretion of DCs. Supported by: Novartis. Disclosure: Dr. Thomas has received research support from Teva Neuroscience. Dr. Sehr has nothing to disclose. Dr. Hainke has nothing to disclose. Dr. Schultheiss has nothing to disclose. Dr. Ziemssen has received personal compensation for activities with Biogen Idec, Bayer Healthcare, Novartis, Sanofi-Aventis Pharmaceuticals, Inc., Teva Neuroscience, and Synthon. Dr. Ziemssen has received research support for from Bayer Healthcare, Biogen Idec, Novartis, Teva Neuroscience, and Sanofi-Aventis Pharmaceuticals, Inc.

Identification of Immunophenotypic Signatures in the Innate Immune System of MS Patients by Multiparametric Flow Cytometry (P02.098)

Objective: Identification of cell surface markers in peripheral blood to distinguish patients from controls. Background MS patients by multiparametric flow cytometry Background: MS is a chronic inflammatory and demyelinating disease of the CNS with unknown etiology until now. CD4+ T helper (Th) 1 cells, proinflammatory Th17 cells, CD8+ T cells, B cells, natural killer (NK) cells and denritic cells (DC) are accepted to play in important role in pathogenesis of disease. Lymphocytes of the peripheral blood from multiple sclerosis (MS) patients are characterized by proinflammatory function but robust cell surface markers to distinguish patients from controls are not available until now. Design/Methods: In this pilot study we analysed phenotypes and their frequencies of peripheral blood cell subsets by multicolour flow cytometry in a limited set of MS patients. The combination of 50 different monoclonal antibodies allowed the detection of nearly 1000 immunophenotypic parameters per sample. Relative and, absolute cell numbers including relative fluorescence intensities of all fluorochromes were compared between 10 patients with early relapsing-remitting MS (RRMS) and 10 healthy age- and sex-matched controls. Results: Immunophenotypic signatures have been identified that allowed a clear classification of MS-patients if compared to healthy controls. These signatures were composed of parameters, which predominantly describe changes in the cells of the innate immune system, such as NK cells, monocytes and neutrophils. For example, in MS IFNg receptor on NK cells, CD62L on granulocytes and CCR2 on neutrophiles was upregulated in MS compared to controls. Conclusions: Our innovative approach identified for the first time MS-specific immunophenotypic signatures of peripheral blood leukocytes by multiparametric flow cytometry. Expression receptors involved in function and activation of innate immune cells showed a significant increase in MS patients compared to controls. Thus, large–scale immunophenotyping is an encouraging tool to identify new disease relevant leukocyte subtypes in a hypothesis-based and hypothesis-generating manner. Supported by: Parts of the study were supported by Merck Serono. Disclosure: Dr. Schulte-Wrede has nothing to disclose. Dr. Grun has nothing to disclose. Dr. Scholz has nothing to disclose. Dr. Harms has received personal compensation for activities with Biomarin, Biogen, Bayer Health Care, Talecris, Merck-Serono as scientific advisory board member and/or speaker. Dr. Grutzkau has nothing to disclose. Dr. Rosche has received personal compensation for activities with Biogen Idec, Bayer-Schering, Teva, and Merck-Serono as a speaker.

Cytokine Patterns After Therapy With Avonex ® , Rebif ® , Betaferon ® and CinnoVex™ in Relapsing–Remitting Multiple Sclerosis in Iranian Patients

Several lines of evidence exist which suggest that changes in the expression of circulating cytokines are linked to the development or reoccurrence of multiple sclerosis (MS). This study aimed to evaluate the serum levels of relevant cytokines after therapy with IFN-β formulations in MS patients. In this study, blood samples were collected from 70 MS patients undergoing four different types of IFN-β formulation treatment and 100 healthy controls. After 24 months of treatment, the serum levels of IL-17A, IL-12, IFN-γ and IL-10 in patients and healthy controls were analyzed by ELISA. Our results demonstrated that serum levels of IL-17A were significantly higher in patients treated with CinnoVex™ and Avonex® when compared with healthy controls. Serum levels of IL-10 were significantly decreased after therapy with CinnoVex, whereas serum levels of IFN-β were elevated. No difference in serum levels of IL-12 were detected between patients and controls. Results of our study suggest that CinnoVex and Avonex modulate the immune system less than Rebif® and Betaferon® in MS patients, and an elevated dose of CinnoVex and Avonex may be required for better regulation of the immune system in MS patients.

MicroRNAs miR-17 and miR-20a Inhibit T Cell Activation Genes and Are Under-Expressed in MS Whole Blood

It is well established that Multiple Sclerosis (MS) is an immune mediated disease. Little is known about what drives the differential control of the immune system in MS patients compared to unaffected individuals. MicroRNAs (miRNAs) are small non-coding nucleic acids that are involved in the control of gene expression. Their potential role in T cell activation and neurodegenerative disease has recently been recognised and they are therefore excellent candidates for further studies in MS. We investigated the transcriptome of currently known miRNAs using miRNA microarray analysis in peripheral blood samples of 59 treatment naïve MS patients and 37 controls. Of these 59, 18 had a primary progressive, 17 a secondary progressive and 24 a relapsing remitting disease course. In all MS subtypes miR-17 and miR-20a were significantly under-expressed in MS, confirmed by RT-PCR. We demonstrate that these miRNAs modulate T cell activation genes in a knock-in and knock-down T cell model. The same T cell activation genes are also up-regulated in MS whole blood mRNA, suggesting these miRNAs or their analogues may provide useful targets for new therapeutic approaches.

Lymphocyte Subpopulations, Oxidative Burst and Apoptosis in Peripheral Blood Cells of Patients with Multiple Sclerosis–Effect of Interferon-β

At present, the most efficient therapeutical treatment of multiple sclerosis (MS) is achieved by IFN-β. However, its in vivo effects remain incompletely understood. If applied parenterally, the hydrophobic IFN-β acts primarily on blood cells with probable selectivity for functionally different lymphocyte subpopulations, monocytes and granulocytes. We have investigated the expression of the activation marker interleukin-2 receptor-α (CD25) on CD3+ T cells, CD19+ B cells, foetal-type γδ+CD3+ T cells and foetal-type CD5+CD19+ B cells of the peripheral blood. In addition, the oxidative burst activity and apoptosis have been determined in mononuclear and polymorphonuclear blood cells, respectively. The study accompanied a phase III trial with IFN-β1b (BETAFERON®, Schering). Two groups of MS patients with relapsing-remitting course of the disease have been investigated at 8 time points (days 0, 5, 15, 31, 60, 90, 180 and 270 after starting therapy): (1) verum group (n=8) with application of 8 Mill. units IFN-β1b every other day, and (2) placebo group (n=4) with application of placebo for 3 months and therapy as in (1) from day 90 onward. The main results were: (1) Activated T cells decreased until day 180 in the verum group and return thereafter to pre-treatment values, whereas in the placebo group the values remained relatively stable over the whole observation period. (2) Activated B cells increased between days 90 and 270 in both groups, i.e. after verum application in both groups. (3) Foetal-type B cells were more activated than total B and T cells with increase over time in both groups. (4) Foetal-type T cells exerted relatively stable intra-individual levels with generally low CD25 expression, but punctual CD25 peaks in both groups. (5) The spontaneous oxidative burst was higher in lymphocytes, more variable in monocytes and faster increasing in granulocytes in the verum group than in the placebo group. (6) Apoptosis of mononuclear cells and granulocytes showed similar variations in the verum and placebo groups with the exception of a selective increase over time of the proportion of granulocytes undergoing induced apoptosis in the verum group. It is concluded that IFN-β has the following main effects on the immune system of MS patients: (1) the T cell immunity is systemically and reversibly suppressed, (2) the foetal-type lymphocytes, which are responsible for the first line of defence of infections, are stimulated in the long range, (3) the oxidative burst activity is increased in lymphocytes and granulocytes and instable in monocytes, and (4) the inducibility of apoptosis in granulocytes is increased. Re-examination of the altered blood cell parameters after long-term IFN-β therapy is warranted.