Concomitant Expression of IL-6 and TGF-β Cytokines and their Receptors in Peripheral Blood of Patients with Multiple Sclerosis: The Effects of INFβ Drugs

BACKGROUND: Concomitant signals from IL-6 and TGF-β have a central role in the Th17 cells development and differentiation, and these cells are the main promoters of demyelinating inflammation in the central nervous system (CNS) resulting in multiple sclerosis (MS). OBJECTIVES: To evaluate the simultaneous IL-6 and TGF-β gene and their receptor protein expression in Relapsing-Remitting (RR)-MS patients. MATERIALS AND METHODS: IL-6 and TGF-β mRNA and their receptor expression on the surface of CD4+T cells were evaluated using real-time PCR (RT-PCR) and flow cytometry, respectively. RESULTS: The IL-6 mRNA expression in patients with RRMS was significantly higher than in the controls (p= 0.019). When patients who did not receive any other treatment were compared with the controls, the significant difference was substantial (p=0.006). The TGF-β mRNA expression in patients was lower than in the controls (p = 0.03). However, in patients receiving IFNβ, it increased compared with the other patients (p= 0.036). There was no difference in cytokine receptor expression between the patients and the control groups. CONCLUSION: Our data conclude an increase and decrease in mRNA expression levels of IL-6 and TGF-β in patients with RRMS, respectively. Moreover, there were no significant differences in receptor expression of either cytokines. Based on our data a balance of TGF and IL-6 appears to have a positive impact on the disease control

Treatment with Rutin - A Therapeutic Strategy for Neutrophil-Mediated Inflammatory and Autoimmune Diseases

Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-in- flammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor (TNF)-α productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin (25 μM) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the TNF-α, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin (1 - 100 μM), after which MTT was appended and incubated at 37ºC for 4 hour. Results: Rutin at concentrations up to 100 μM did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and TNF-α productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and TNF-α productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/ autoimmune diseases

Therapeutic plasma exchange may adjust IL-6 and TGF-β signals in relapsed MS patients peripheral blood

Objective: Effects of therapeutic plasma exchange (TPE) on immune cells and their cytokine production in MS, are unknown. Since interleukine-6 and tumor growth factor-β have critical roles in MS immunopathogenesis, the impacts of TPE on the expression of these cytokines and their receptors on the surface of CD4+ T lymphocytes, were investigated. Methods: Blood cells were obtained from 30 Relapsing-Remitting (RR) MS patients, before and after a complete TPE course. Cytokines mRNA and their receptor expression on the CD4+ T cells surface were assessed using real-time PCR and flowcytometry, respectively. Results: TPE reduced symptom severity (P =.01) and the relief was higher in males than in females (P =.039). TPE also increased TGF-β mRNA and decreased IL-6 receptor expressing cells frequency (P =.009 and P =.028, respectively). Moreover, the frequency of CD4+IL6R+ T cells was positively correlated with disease severity (P =.001). Conclusion: TPE impacts simultaneously on the TGF-β mRNA and IL-6 receptor expression, and this may be a mechanism of improvement in MS relapse symptoms induced by the TPE

Detection of Interleukin-19 mRNA in C57BL/6 Mice Astroglial Cells and Brain Cortex

Introduction: Astrocytes are the most abundant glial cell type. In addition to their neurological roles, astrocytes also have immune functions. They have been involved in antigen presentation in the central nervous system (CNS). Activated astrocytes express adhesion molecules, chemokines and release several inflammatory mediators, pro-inflammatory cytokines, neurotrophic and neuroprotective factors, thus these cells have a dual role within the CNS: neuroinflammation and repair processes. IL-19, IL-20, IL-22, IL-24, IL-26, IL-28A, IL-28B, and IL-29 are members of the IL-10 family of cytokines. These cytokines have different biological functions in spite of partial amino acid sequences homology. Signal transduction of the IL-10 family of cytokines is through R1-type and R2-type receptors. Methods: No information has been available about the expression and regulation of IL-19 in mice astrocytes and brain. To investigate the expression of IL-19, we examined its expression in C57BL/6 mice astroglial cells in response to lipopolysaccharide (LPS), using reversetranscription polymerase chain reaction (RT-PCR) method.  Results: We provide for the first time, evidence that astrocytes can express IL-19 mRNA following LPS stimulation. Furthermore, we have found the expression of IL-19 mRNA in the cortex of adult C57BL/6 mice following intraperitoneal (i.p.) administration of LPS.  Discussion: This finding will contribute to current knowledge on the function and behavior of cells and mediators during inflammatory conditions in the brai

Therapeutic efficacy of Urtica dioica and evening primrose in patients with rheumatoid arthritis: A randomized double-blind, placebo-controlled clinical trial

Background: Rheumatoid arthritis (RA) is a systemic inflammatory autoimmune disease. The pharmacological therapy of RA is often symptomatic to mitigate pain and immobility with analgesics and drugs with defined side effects and risks. Complementary medicines might decrease the signs of RA and reduce the need for them. In the present study, the anti-rheumatic, anti-inflammatory, and anti-oxidant effects of Urtica dioica and Evening Primrose Oil (EPO), in patients with RA was investigated. Methods: This randomized, double‐blind, controlled trial involved 90 RA patients, and randomly assigned them into EPO, Urtica dioica, and placebo groups. The potential effect of these herbal medicines on Disease Activity Score (DAS) 28, Visual Analogue Scale (VAS), Total Anti-oxidant Capacity (TAC), IL-17, Rheumatoid Factor (RF), anti-cyclic citrullinated peptide antibodies (Anti-CCP), C Reactive Protein (CRP), and Erythrocyte Sedimentation Rate (ESR) before and after clinical trial were evaluated. Results: After a three-month follow-up, the mean values of DAS28, IL-17, TAC, RF, and CRP in EPO and Urtica dioica groups were significantly different from the placebo group. However, the VAS, Anti-CCP, and ESR at baseline and at the end of the study were not significantly different between the three groups. After the intervention, the within-group DAS28 in the EPO and Urtica dioica groups, and placebo groups reduced significantly compared to the baseline. Conclusion: Medicinal plants EPO and Urtica dioica appeared to decrease inflammatory factors, and can improve the symptoms of RA. Thus, EPO and Urtica dioica have great potential as complementary therapy in RA patients

Induced Pluripotent Stem Cell Meets Severe Combined Immunodeficiency

Severe combined immunodeficiency (SCID) is classified as a primary immunodeficiency, which is characterized by impaired T-lymphocytes differentiation. IL2RG, IL7Ralpha, JAK3, ADA, RAG1/RAG2, and DCLE1C (Artemis) are the most defective genes in SCID. The most recent SCID therapies are based on gene therapy (GT) of hematopoietic stem cells (HSC), which are faced with many challenges. The new studies in the field of stem cells have made great progress in overcoming the challenges ahead. In 2006, Yamanaka et al. achieved "reprogramming" technology by introducing four transcription factors known as Yamanaka factors, which generate induced pluripotent stem cells (iPSC) from somatic cells. It is possible to apply iPSC-derived HSC for transplantation in patients with abnormality or loss of function in specific cells or damaged tissue, such as T-cells and NK-cells in the context of SCID. The iPSC-based HSC transplantation in SCID and other hereditary disorders needs gene correction before transplantation. Furthermore, iPSC technology has been introduced as a promising tool in cellular-molecular disease modeling and drug discovery. In this article, we review iPSC-based GT and modeling for SCID disease and novel approaches of iPSC application in SCID