transforming-growth-factor-beta and Multiple-Sclerosis

Multiple sclerosis (MS) is an acute demyelinating disease with an autoimmune nature, followed by gradual neurodegeneration and enervating scar formation. Dysregulated immune response is a crucial dilemma contributing to the pathogenesis of MS. The role of chemokines and cytokines, such as transforming growth factor-β (TGF-β), have been recently highlighted regarding their altered expressions in MS. TGF-β has three isoforms, TGF-β1, TGF-β2, and TGF-β3, that are structurally similar; however, they can show different functions.. All three isoforms are known to induce immune tolerance by modifying Foxp3. To develop novel neuroimmunological treatment strategies for MS, the optimal strategy could be the one that causes immune modulation, induces neurogenesis, stimulates remyelination, and prevents excessive scar formation. Therefore, regarding its immunological properties, TGF-β could be an appropriate candidate; however, contradictory results of previous studies have questioned its role and therapeutic potential in MS. In this review article, we provide an overview of the role of TGF-β in immunopathogenesis of MS, related clinical and animal studies, and the treatment potential of TGF-β in MS, emphasizing the role of different TGF-β isoforms.

Topics: Animals; Cicatrix; Humans; Multiple Sclerosis; Protein Isoforms; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transforming Growth Factor beta3

Controlling CD4

Topics: Blood-Brain Barrier; Cell Differentiation; Humans; Multiple Sclerosis; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

Recent advances in understanding the pathogenesis of multiple sclerosis (MS) have brought into the spotlight the major role played by reactive astrocytes in this condition. Response Gene to Complement (RGC)-32 is a gene induced by complement activation, growth factors, and cytokines, notably transforming growth factor β, that is involved in the modulation of processes such as angiogenesis, fibrosis, cell migration, and cell differentiation. Studies have uncovered the crucial role that RGC-32 plays in promoting the differentiation of Th17 cells, a subtype of CD4

Topics: Animals; Biomarkers; Cell Cycle Proteins; Complement System Proteins; Cytokines; Humans; Mice; Multiple Sclerosis; Muscle Proteins; Nerve Tissue Proteins; Neuroinflammatory Diseases; Nuclear Proteins; Transforming Growth Factor beta

Bone Morphogenetic Proteins (BMP) and Transforming Growth Factor-beta (TGF-β) are cytokines with similar receptors and messengers. They are important for immune cell function, with BMPs exerting mainly proinflammatory but also anti-inflammatory effects, and TGF-β suppressing inflammation. Patients with Multiple Sclerosis exhibit BMP overactivity and suppressed TGF-β signaling. This dysregulated signaling participates in the crosstalk between infiltrating immune cells and glia, where BMP inhibits remyelination. Reciprocal antagonism between the two pathways takes place via a variety of mechanisms. Although this antagonism has not been studied in the setting of Multiple Sclerosis, it could inform further research and treatment discovery.

Topics: Animals; Bone Morphogenetic Proteins; Humans; Multiple Sclerosis; Signal Transduction; Transforming Growth Factor beta

Transforming growth factor beta (TGF-β) is a pleiotropic cytokine that has been shown to influence the differentiation and function of T cells. The role that TGF-β plays in immune-mediated disease, such as multiple sclerosis (MS), has become a major area of investigation since CD4

Topics: Animals; Autoantigens; Autoimmunity; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Humans; Mice; Multiple Sclerosis; Signal Transduction; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Multiple sclerosis is the most frequent chronic inflammatory, demyelinating and neurodegenerative disease in young adults, but has no definitive pharmacological treatment. It is a heterogeneous disease from the immunological, neuropathological and clinical point of view, as well as in terms of its response to different therapies. Over the last two decades, pharmacology has focused on developing drugs that are capable of modifying the course of this disease, with the aim of reducing the frequency of the outbreaks and the speed at which the disability produced by the disease progresses. Nevertheless, today, there are no drugs that are capable of offering a curative effect that can fully stabilise the disease, and neuroprotective and neuroreparative strategies are still in their early stages. In this work we carry out a critical review of the different pathogenic paths involved in multiple sclerosis and we discuss the different pharmacological approaches that have been followed, based on the clinical trials that are currently being conducted. In the near future it is to be expected that, first, we will manage to stabilise the disease completely and, later, recover some of the functions altered by this disease. Research is being conducted at such a rate that we have to be optimistic and think that soon we will be able to improve the situation of those who suffer from the disease.

Topics: Adenosine Deaminase; Antigen-Presenting Cells; Antigens, CD; Cell Adhesion Molecules; Dihydroorotate Dehydrogenase; Humans; Lysophospholipids; Matrix Metalloproteinases; Multiple Sclerosis; Myelin Basic Protein; NF-kappa B; Oxidoreductases Acting on CH-CH Group Donors; Sphingosine; Transforming Growth Factor beta

Transforming growth factor beta (TGF-β) has a crucial role in the differentiation of ectodermal cells to neural or epidermal precursors. TGF-β and bone morphogenetic protein molecules (BMPs) are involved in many developmental processes, including cell proliferation and differentiation, apoptosis, mitotic arrest and intercellular interactions during morphogenesis. Additionally, the failure of central thymic tolerance mechanisms, leading to T cells with a skewed autoreactive response, is being described as a contributor in inflammatory processes in autoimmune diseases such as multiple sclerosis. Since TGF-β and BMP proteins are crucial for the development of the neural system and the thymus, as well as for the differentiation of T cells, it is essential to further investigate their role in the pathophysiology of this disorder by using references from embryonic experimental research. Available literature in the TGF/BMP signalling cascade, mostly during embryonic development of the nervous system is being reviewed. An attempt is made to further elucidate a potential role of TGF/BMP signalling in the pathophysiology of MS. During demyelination, BMP signaling, through various molecular mechanisms, directs the development of the adult neural stem cell in the astrocyte rather than the oligodendrocyte direction, therefore inhibiting the repair process. Further understanding of the above relationships could lead to the development of potentially efficient therapies for MS in the future.

Topics: Animals; Autoimmune Diseases; Bone Morphogenetic Proteins; Epidermis; Humans; Multiple Sclerosis; Nervous System; Nervous System Diseases; Smad Proteins; Transforming Growth Factor beta

Multiple sclerosis (MS) is a CD4+ T cell-mediated autoimmune disease affecting the central nervous system. It was largely accepted that Th1 cells driven by IL-12 were pathogenic T cells in human MS and experimental autoimmune encephalomyelitis, an animal model of MS. Recent data have established that IL-17-producing CD4+ T cells, driven by IL-23 and referred to as Th17 cells, play a pivotal role in the pathogenesis of EAE. A combination of TGF-beta and IL-6 induce Th17 cell lineage commitment via expression of transcription factor RORgammat. Th17 cells and induced Foxp3+ T regulatory cells are in reciprocal position in the T cell lineage commitment governed by TGF-beta and IL-6. The vitamin A metabolite retinoic acid is involved in this process via TGF-beta dependent induction of Foxp3. We have demonstrated that human Th17 cells could be identified as CCR2+ CCR5- memory CD4+ T cells. It is becoming clear that IL-23/Th17 axis also plays an important role in the pathogenesis of various human autoimmune diseases including MS. Additionally, accumulating evidences raise a possibility that CCR2 on Th17 cells may be a therapeutic target in MS.

Topics: Animals; Autoimmune Diseases; Cell Differentiation; Encephalomyelitis; Encephalomyelitis, Autoimmune, Experimental; Forkhead Transcription Factors; Humans; Interleukins; Multiple Sclerosis; Nuclear Receptor Subfamily 1, Group F, Member 3; Receptors, CCR2; Receptors, Retinoic Acid; Receptors, Thyroid Hormone; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer; Th1 Cells; Transforming Growth Factor beta

Mesenchymal stem cells possess the ability to differentiate into osteoblasts, chondroblasts, lipoblasts, myoblasts and so on, which can be used in the formation of hematopoietic microenvironment, tissue repairing and gene therapy. Growth factors such as TGF-beta, IGF-I, BMP and FGF can influence on the differentiation of MSC and they cooperate with each other. MSCs support hematopoiesis by secreting cytokines including G-CSF, SCF, LIF, M-CSF, IL-6, IL-11 and are related to some diseases. MSC would demonstrate important effect on gene engineering.

Topics: Animals; Bone Morphogenetic Proteins; Cytokines; Fibroblast Growth Factors; Genetic Engineering; Humans; Mesoderm; Multiple Sclerosis; Osteoporosis; Stem Cells; Transforming Growth Factor beta

The use of transgenic technology to over-express or prevent expression of genes encoding molecules related to inflammation has allowed direct examination of their role in experimental disease. This article reviews transgenic and knockout models of CNS demyelinating disease, focusing primarily on the autoimmune disease multiple sclerosis, as well as conditions in which an inflammatory response makes a secondary contribution to tissue injury or repair, such as neurodegeneration, ischemia and trauma.

Topics: Animals; Central Nervous System; Chemokines; Clinical Trials as Topic; Cytokines; Demyelinating Diseases; Humans; Interferon-gamma; Models, Genetic; Multiple Sclerosis; Th2 Cells; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Inflammatory demyelinating diseases comprise a heterogeneous group of disorders that affect the peripheral and central nervous system. Multiple sclerosis (MS) is the most common disease affecting the CNS white matter. Close similarities between MS and the animal model of the disease, experimental allergic encephalitis (EAE), have suggested that MS might be an autoimmune disease, which is triggered by an infectious agent. Our laboratory has directed its effort in identifying and designing therapies that interfere with key signaling pathways that mediate CNS inflammation in experimental allergic encephalitis. These have included naturally occurring cytokines such as TGFbeta and synthetic small molecules, lysofyline and tyrphostin, which inhibit the inflammatory response and prevent the development of EAE.

Topics: Adult; Animals; Anti-Inflammatory Agents, Non-Steroidal; CD4-Positive T-Lymphocytes; Cerebrospinal Fluid; Chlamydia Infections; Chlamydophila pneumoniae; Cytokines; Demyelinating Autoimmune Diseases, CNS; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Enzyme Inhibitors; Glatiramer Acetate; Humans; Immunotherapy; Inflammation; Interleukin-12; Mice; Multiple Sclerosis; Pentoxifylline; Peptides; Protein Kinase Inhibitors; Protein Kinases; Signal Transduction; Transforming Growth Factor beta

IFN beta-1b reduces the frequency of major multiple sclerosis attacks by 50 percent. Serial MRI scanning over the course of the clinical trial that led to approval of the agent revealed a significant lessening both in disease activity and in accumulating burden of disease in IFN beta-1b-treated patients compared to placebo-treated controls. The mechanism by which IFN beta-1b exerts its beneficial effect in multiple sclerosis is unknown. T suppressor cell function fails during MS attacks and is persistently subnormal in multiple sclerosis patients with progressive disease. IFN beta-1b partially restores suppressor function in multiple sclerosis patients. IFN beta-1b also inhibits release of lymphotoxin, tumor necrosis factor, and interferon gamma, at least in vitro. All three cytokines are toxic to oligodendrocytes. In contrast; production of transforming growth factor beta-1 (TGF beta 1) is increased by IFN beta-1b. TGF beta 1 is an immunosuppressive cytokine. All of the above listed actions of IFN beta-1b could contribute to its beneficial effect. Perhaps all do.

Topics: Demyelinating Diseases; Humans; Interferon beta-1a; Interferon beta-1b; Interferon-beta; Interferon-gamma; Magnetic Resonance Imaging; Multiple Sclerosis; Recombinant Proteins; Transforming Growth Factor beta

The Transforming Growth Factor-betas (TGF-beta) are a group of multifunctional proteins whose cellular sites of production and action are widely distributed throughout the body, including the central nervous system (CNS). Within the CNS, various isoforms of TGF-beta are produced by both glial and neural cells. When evaluated in either cell culture or in vivo models, the various isoforms of TGF-beta have been shown to have potent effects on the proliferation, function, or survival of both neurons and all three glial cell types, astrocytes, microglia and oligodendrocytes. TGF-beta has also been shown to play a role in several forms of acute CNS pathology including ischemia, excitotoxicity and several forms of neurodegenerative diseases including multiple sclerosis, Parkinson's disease, AIDS dementia and Alzheimer's disease.

Topics: AIDS Dementia Complex; Alzheimer Disease; Animals; Astrocytes; Central Nervous System; Encephalomyelitis, Autoimmune, Experimental; Gene Expression; Humans; Ischemia; Microglia; Multiple Sclerosis; Neurodegenerative Diseases; Oligodendroglia; Parkinson Disease; Transforming Growth Factor beta

Topics: alpha-Fetoproteins; Animals; Autoimmune Diseases; Autoimmunity; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Guinea Pigs; Humans; Immune Tolerance; Multiple Sclerosis; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Puerperal Disorders; Rabbits; Rats; Recurrence; Retrospective Studies; Species Specificity; Transforming Growth Factor beta

Management of multiple sclerosis (MS) is based on the usage of immunosuppressive and immune-modulating medications. Cytokines play an important role in the pathogenesis of MS.. To evaluate the effects of rapamycin on the concentrations of Th1/Th2/Th17 serum cytokines in patients with MS.. Six patients with relapsing remitting MS as a case group and 6 healthy individuals as a control group were enrolled. The patients have been receiving 2 mg rapamycin daily for 6 months. The individuals in control group received nothing during 6 months of the experiment. Enzyme linked immunosorbent assay (Simultaneous Multi-Analyte ELISA) technique was used for determination of serum concentrations of IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, IFN-γ, TNF-α, G-CSF and TGF-β before and after therapy with rapamycin.. The mean absorbance of 10 out of the 12 studied cytokines showed reduction after the therapy with rapamycin including IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, IFN-γ and TNF-α. The only statistically significant reduction was observed in the absorbance of IFN-γ (p=0.028). Two cytokines illustrated increase in the patients sera after the therapy, including G-CSF and TGF-β, but only increase in TGF-β was statistically significant (p=0.046). None of the studied cytokines in the control group varied significantly after 6 months.. Based on the findings of this study, rapamycin has some immunosuppressive effects, such as decreasing IFN -γ, which can improve the quality of life of the patients with multiple sclerosis. Also the increased level of TGF-β may also have benefits on the disease, which needs further clinical studies.

Topics: Adult; Enzyme-Linked Immunosorbent Assay; Female; Granulocyte Colony-Stimulating Factor; Humans; Interferon-gamma; Iran; Male; Multiple Sclerosis; Sirolimus; Th1 Cells; Th17 Cells; Th2 Cells; Transforming Growth Factor beta; Young Adult

Deficiency of vitamin D is an environmental risk factor for MS. Vitamin D has immunomodulatory effects, including promotion of T-cell differentiation into T-regulatory cells, which produces regulatory cytokines including TGF-β. Increasing serum vitamin D levels have been associated with decreased disease activity in MS patients, but there are only few studies concerning the immunological effects of vitamin D supplementation in MS. In this study we investigated the effect of weekly supplementation of vitamin D3 or placebo on serum levels of multiple cytokines in patients with relapsing remitting MS.. The study was conducted on the patient cohort of the Finnish Vitamin D study. All patients were using IFN-beta-1b and were randomized to add-on treatment with either cholecalciferol 20,000 IU/week or placebo. Concentrations of LAP (TGF-β), INF-γ, IL-17A, IL-2, IL-10, IL-9, IL-22, IL-6, IL-13, IL-4, IL-5, IL-1β and TNF-α were determined at screening and at 12 months using commercial fluorescent bead immunoassay kits.. LAP (TGF-β) levels increased significantly in the vitamin D treated group from a mean of 47 (SE 11) pg/ml to 55 (SE 14) pg/ml in 12 months (p-value=0.0249). Placebo treatment had no significant effect on LAP levels. The levels of the other cytokines did not change significantly in either group.. We showed increased serum latency activated peptide (LAP) of TGF-β levels in MS patients treated with vitamin D3. The immune regulatory effects of TGF-beta may play a role in the improved MRI outcomes that we observed earlier in the vitamin D treated group of patients.

Topics: Adult; Cytokines; Double-Blind Method; Female; Follow-Up Studies; Humans; Major Histocompatibility Complex; Male; Middle Aged; Multiple Sclerosis; Time Factors; Transforming Growth Factor beta; Vitamin D; Vitamins; Young Adult

Multiple sclerosis (MS) is an inflammatory disease in which the myelin sheaths around the axons of the central nervous system are damaged. The damage leads to demyelination and scarring as well as a broad spectrum of signs and symptoms. The epidemiological data suggest a possible influence of vitamin D as an immunomodulatory agent on multiple sclerosis susceptibility as well as on clinical course of the disease. We investigated the effects of short-term vitamin D3 therapy on Iranian patients with MS. In a prospective randomized controlled trial study, 62 MS patients received 300,000 IU/month vitamin D3 or placebo as intramuscular injection for 6 months. Our results showed no significant difference between the treatment and the control groups in the expanded disability status scale scores and number of gadolinium-enhancing lesions during the 6-month treatment period. After 6 months, the levels of cell proliferation in the vitamin D treatment group were significantly lower than the control group. Also, the levels of transforming growth factor-beta and interleukin-10 in the vitamin D treatment group were significantly higher than the control group. This result suggests that vitamin D therapy may help prevent the development of MS and could be a useful addition to the therapy.

Topics: Adolescent; Adult; Calcitriol; Cell Proliferation; Cholecalciferol; Double-Blind Method; Female; Humans; Injections, Intramuscular; Interferon-gamma; Interleukin-10; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Middle Aged; Multiple Sclerosis; Phytohemagglutinins; T-Lymphocytes; Transforming Growth Factor beta; Treatment Outcome; Young Adult

To assess whether TGF-beta isoforms are significantly increased after intravenous immunoglobulin (i.v.IG) infusion in the plasma of patients with multiple sclerosis (MS), 19 patients with clinically definite MS were enrolled in a double blind placebo controlled i.v.IG study. TGF-beta1, TGF-beta2, TGF-beta3 plasma concentrations were measured prior and directly after i.v.IG infusions by specific ELISA. Compared to the placebo group, we found a significant increase in the plasma levels of all three TGF-beta isoforms in patients treated with i.v.IG. The significantly increased TGF-beta plasma concentrations in treated patients suggest an additional, immediate mechanism of action that may accompany the molecular effects of i.v.IG therapy in MS. The variable amount of the potent anti-inflammatory TGF-beta isoforms within the i.v.IG preparations may exert a differentiated view regarding the manifold indications of i.v.IG therapy.

Topics: Double-Blind Method; Enzyme-Linked Immunosorbent Assay; Humans; Immunoglobulins, Intravenous; Multiple Sclerosis; Protein Isoforms; Transforming Growth Factor beta

Multiple sclerosis (MS) patients were randomized, in a double blind design, and placed into either a vitamin D supplemented group or a placebo control group. As expected, serum 25-hydroxyvitamin D levels increased significantly following 6 month vitamin D supplementation (17+/-6 ng/ml at baseline to 28+/-8 ng/ml at 6 months). Vitamin D supplementation also significantly increased serum transforming growth factor (TGF)-beta 1 levels from 230+/-21 pg/ml at baseline to 295+/-40 pg/ml 6 months later. Placebo treatment had no effect on serum TGF-beta 1 levels. Tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and interleukin (IL)-13 were not different following vitamin D supplementation. IL-2 mRNA levels decreased following vitamin D supplementation but the differences did not reach significance. Vitamin D supplementation of MS patients for 6 months was associated with increased vitamin D status and serum TGF-beta 1.

Topics: Cytokines; Female; Humans; Immune System; Immune Tolerance; Inflammation Mediators; Interferon-gamma; Interleukin-13; Interleukin-2; Male; Multiple Sclerosis; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1; Treatment Outcome; Tumor Necrosis Factor-alpha; Vitamin D; Vitamin D Deficiency

Transforming growth factor (TGF)-beta2 is a pleiotropic cytokine associated with remissions in multiple sclerosis (MS) and amelioration of allergic encephalomyelitis. We assessed the safety of TGF-beta2 in an open-label trial of 11 patients with secondary progressive (SP) MS. Five patients had a reversible decline in the glomerular filtration rate. There was no change in expanded disability status scale or MRI lesions during treatment. Systemic TGF-beta2 may be associated with reversible nephrotoxicity, and further investigation of its therapeutic potential in MS should be performed with caution.

Topics: Adult; Blood Urea Nitrogen; Cerebrospinal Fluid; Chronic Disease; Female; Glomerular Filtration Rate; Humans; Male; Middle Aged; Multiple Sclerosis; Renal Circulation; Transforming Growth Factor beta

The serum levels of TGF-beta1, measured by solid-phase ELISA, were determined to be significantly augmented in patients with both relapsing remitting (RR) and secondary chronic progressive (CP) MS compared with sex- and age-matched healthy controls. Moreover, in RR MS patients, the blood levels of the cytokine were further augmented either during relapses or, in a rapid but reversible fashion, by s.c. injection with 8 million International Units (MIU) IFN-beta1b. Because TGF-beta1 possesses multiple anti-inflammatory activities, we hypothesize that the increase in its circulating levels in RR and CP MS patients might represent an endogenous anti-inflammatory mechanism aimed at counteracting ongoing immunoinflammatory events, and that IFN-beta may further potentiate this natural defensive apparatus.

Topics: Adjuvants, Immunologic; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interferon beta-1a; Interferon beta-1b; Interferon-beta; Male; Multiple Sclerosis; Recombinant Proteins; Recurrence; Transforming Growth Factor beta

Oral administration of antigen is a long-recognized method of inducing systemic immune tolerance. In animals with experimental autoimmune disease, a major mechanism of oral tolerance involves the induction of regulatory T cells that mediate active suppression by secreting the cytokine TGF-beta 1. Multiple sclerosis (MS) is a presumed T cell-mediated Th1 type autoimmune disease. In this paper we investigated, in patients with MS, whether oral myelin treatment (myelin containing both MBP and PLP) induced antigen-specific MBP- or PLP-reactive T cells that were either Th2-like (secreted IL-4 or TGF-beta 1), or alternatively whether Th1 type sensitization occurred as measured by IFN-gamma secretion. Specifically, 4,860 short-term T cell lines were generated to either MBP, PLP or TT from 34 relapsing-remitting patients with MS; 17 were orally treated with bovine myelin daily for a minimum of two years as compared to 17 non-treated patients. We found a marked increase in the relative frequencies of both MBP- and PLP-specific TGF-beta 1 secreting T cell lines in the myelin-treated MS patients as compared to non-treated MS patients (MBP, p < 0.001; PLP, p < 0.003). In contrast, no changes in the frequency of MBP- or PLP-specific IFN-gamma or TT-specific TGF-beta 1 secreting T cells were observed. These results suggest that the oral administration of antigens generates antigen-specific TGF-beta 1 secreting T cells of presumed mucosal origin that may represent a distinct cytokine-secreting lineage of T cells (Th3). Since, in animal models, antigen-specific TGF-beta 1 secreting cells localize to the target organ and then suppress inflammation in the local microenvironment, oral tolerization with self-antigens may provide a therapeutic approach for the treatment of cell-mediated autoimmune disease which does not depend upon knowledge of the antigen specificity of the original T cell clone triggering the autoimmune cascade.

Topics: Administration, Oral; Adult; Antigens; Female; Humans; Immune Tolerance; Interferon-gamma; Interleukin-4; Lymphocyte Activation; Multiple Sclerosis; Myelin Basic Protein; Myelin Proteolipid Protein; T-Lymphocytes; Transforming Growth Factor beta

Oral administration of antigen is a long recognized method of inducing systemic immune tolerance. In animals with experimental autoimmune disease, a major mechanism of oral tolerance triggered by oral administration of antigen involves the induction of regulatory T cells that mediate active suppression by secreting the cytokine TGF-beta 1. Multiple sclerosis (MS) is a presumed T cell-mediated Th1 type autoimmune disease. Here, we investigated whether in MS patients oral myelin treatment, containing both myelin basic protein (MBP) and proteolipid protein (PLP), induced antigen specific MBP or PLP reactive T cells that either secreted IL4, TGF-beta1, or alternatively did Th1 type sensitization occur as measured by IFN-gamma secretion. Specifically, 4,860 short-term T cell lines were generated to either MBP, PLP, or tetanus toxoid (TT) from 34 relapsing-remitting MS patients: 17 orally treated with bovine myelin daily for a minimum of 2 yr as compared to 17 nontreated patients. We found a marked increase in the relative frequencies of both MBP and PLP specific TGF-beta1-secreting T cell lines in the myelin treated MS patients as compared to non-treated MS patients (MBP P < 0.001, PLP P < 0.003). In contrast, no change in the frequency of MBP or PLP specific IFN-gamma or TT specific TGF-beta1 secreting T cells were observed. These results suggest that the oral administration of antigens generates antigen specific TGF-beta1 secreting Th3 cells of presumed mucosal origin that represent a distinct lineage of T cells. Since antigen-specific TGF-beta1 secreting cells localize to the target organ and then suppress inflammation in the local microenvironment, oral tolerization with self antigens may provide a therapeutic approach for the treatment of cell-mediated autoimmune disease which does not depend upon knowledge of the antigen specificity of the original T cell clone triggering the autoimmune cascade.

Topics: Administration, Oral; Autoantigens; Clinical Trials as Topic; Follow-Up Studies; Humans; Immune Tolerance; Interferon-gamma; Interleukin-4; Multiple Sclerosis; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Recurrence; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

T cells play an important role in the development and progression of multiple sclerosis (MS), an autoimmune disease of the central nervous system. In the present study, the immunomodulatory impacts of two Lactobacillus strains, L paracasei DSM 13434 and L plantarum DSM 15312, on the frequency and cytokine production of CD4+ T cells in MS patients were explored. Thirty MS patients were enrolled in this study. The CD4+ T cells were isolated, cultured, and exposed to the media containing cell-free supernatants of L plantarum (group1), L paracasei (group 2), the mixture group of cell-free supernatants of both probiotics (group 3), and vehicle (control) group (group 4). The frequencies of T helper (Th) 1, Th17, Th2, and T regulatory type 1 (Tr1) cells and mean fluorescent intensity (MFI) of the associated cytokines were assessed using flow cytometry. The levels of interleukin 17 (IL-17), transforming growth factor β (TGF-β), and interferon-gamma (IFN-γ) cytokines in supernatants of all groups were measured by enzyme-linked immunosorbent assay. The percentage of Th1 cells and the MFI of IFN-γ in Th1 cells (CD4+ IFN-γ+) in all three probiotic treatment groups were significantly decreased compared to the control group. However, no significant changes were observed in the proportion and MFI of Th2, Th17, and Tr1 cells. A significant decrease was observed in IL-17 secretion in the supernatant of cultured CD4+ T cells in all three treatment groups in comparison with control. The levels of TGF-β and IFN-γ were not significantly different among any of the study groups.  Collectively, cell-free supernatants of the lactobacilli showed an in vitro anti-inflammatory effect. However, further studies are needed to prove the real effects of probiotics on MS.

Topics: CD4-Positive T-Lymphocytes; Cytokines; Humans; Interferon-gamma; Interleukin-17; Lacticaseibacillus paracasei; Lactobacillus; Lactobacillus plantarum; Multiple Sclerosis; Probiotics; Th1 Cells; Transforming Growth Factor beta

Vitamin D is an environmental factor related to multiple sclerosis that plays a significant role in immune regulation. TGF-β is a superfamily of cytokines with an important dual effect on the immune system. TGF-β inhibits the Th1 response while facilitating the preservation of regulatory T cells (FOXP3+) in an immunoregulatory capacity. However, when IL-6 is present, it stimulates the Th17 response. Our aim was to analyze the regulatory effect of vitamin D on the in vivo TGF-β signaling pathway in patients with relapsing-remitting multiple sclerosis (RRMS). A total of 21 patients with vitamin D levels < 30 ng/mL were recruited and supplemented with oral vitamin D. All patients were receiving disease-modifying therapy, with the majority being on natalizumab. Expression of

Topics: Humans; Multiple Sclerosis; Multiple Sclerosis, Relapsing-Remitting; Natalizumab; Transforming Growth Factor beta; Vitamin D; Vitamins

Multiple sclerosis (MS) is a chronic inflammatory and autoimmune disease affecting various inflammatory and nutritional parameters. Therefore, this study aimed to investigate the relationship between the Body Mass Index (BMI) of MS patients and the serum levels of leptin, orexin-A, and Transforming Growth Factor β (TGF-β).. This cross-sectional study included 25 patients suffering from MS and 40 healthy individuals as the case and control groups, respectively. The serum levels of leptin, orexin-A, and TGF-β were assessed in the participants using the Enzyme-Linked Immunosorbent Assay methods. Moreover, data were analyzed using the descriptive statistical indices, t-test, chi-square test, and linear regression test.. According to our results, the participants' mean age was 38.04 ± 7.53 and 40.23 ± 5.88 in the case and control groups, respectively. Also, the groups were not significantly different in gender, age, alcohol consumption, and smoking (p > 0.05). It was found that the mean serum levels of orexin-A and TGF-β were significantly lower in the MS patients compared to the control group, while the mean serum leptin levels were significantly higher (42.8 vs. 18.9 ng/ml, p < 0.001). Moreover, there was no significant relationship between the BMI of the MS patients and their serum levels of orexin-A, TGF-β, and leptin (p > 0.05).. In conclusion, we found significantly lower levels of orexin-A and TGF-β and a significantly higher level of leptin in the MS patients compared to the control group. In addition, there was no significant relationship between the BMI and the serum levels of orexin-A, TGF-β, and leptin in MS patients.

Topics: Adult; Body Mass Index; Cross-Sectional Studies; Female; Humans; Leptin; Lipids; Male; Middle Aged; Multiple Sclerosis; Orexins; Transforming Growth Factor beta

Topics: Brain; DNA Methylation; Humans; Inflammation; Microglia; Multiple Sclerosis; Neurodegenerative Diseases; Transforming Growth Factor beta; White Matter

Topics: Animals; Corpus Callosum; Cuprizone; Demyelinating Diseases; Disease Models, Animal; Interleukin-10; Mesenchymal Stem Cells; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Myelin Sheath; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

In a substantial share of patients suffering from multiple sclerosis (MS), neurological functions slowly deteriorate despite a lack of radiological activity. Such a silent progression, observed in either relapsing-remitting or progressive forms of MS, is driven by mechanisms that appear to be independent from plaque activity. In this context, we previously reported that, in the spinal cord of MS patients, periplaques cover large surfaces of partial demyelination characterized notably by a transforming growth factor beta (TGF-beta) molecular signature and a decreased expression of the oligodendrocyte gene

Topics: Brain; Humans; Multiple Sclerosis; Myelin Sheath; Neoplasm Recurrence, Local; Plaque, Amyloid; RNA, Messenger; Transforming Growth Factor beta

Th17 cells are known to exert pathogenic and non-pathogenic functions. Although the cytokine transforming growth factor β1 (TGF-β1) is instrumental for Th17 cell differentiation, it is dispensable for generation of pathogenic Th17 cells. Here, we examined the T cell-intrinsic role of Activin-A, a TGF-β superfamily member closely related to TGF-β1, in pathogenic Th17 cell differentiation. Activin-A expression was increased in individuals with relapsing-remitting multiple sclerosis and in mice with experimental autoimmune encephalomyelitis. Stimulation with interleukin-6 and Activin-A induced a molecular program that mirrored that of pathogenic Th17 cells and was inhibited by blocking Activin-A signaling. Genetic disruption of Activin-A and its receptor ALK4 in T cells impaired pathogenic Th17 cell differentiation in vitro and in vivo. Mechanistically, extracellular-signal-regulated kinase (ERK) phosphorylation, which was essential for pathogenic Th17 cell differentiation, was suppressed by TGF-β1-ALK5 but not Activin-A-ALK4 signaling. Thus, Activin-A drives pathogenic Th17 cell differentiation, implicating the Activin-A-ALK4-ERK axis as a therapeutic target for Th17 cell-related diseases.

Topics: Activin Receptors, Type I; Activins; Animals; Cell Differentiation; Cells, Cultured; Encephalomyelitis, Autoimmune, Experimental; Humans; Mice; Mice, Knockout; Molecular Targeted Therapy; Multiple Sclerosis; Neurogenic Inflammation; Signal Transduction; Th17 Cells; Transforming Growth Factor beta

Early innate education of hematopoietic progenitors within the bone marrow (BM) stably primes them for either trained immunity or instead immunoregulatory functions. We herein demonstrate that in vivo or in vitro activation within the BM via Toll-like receptor-9 generates a population of plasmacytoid dendritic cell (pDC) precursors (CpG-pre-pDCs) that, unlike pDC precursors isolated from PBS-incubated BM (PBS-pre-pDCs), are endowed with the capacity to halt progression of ongoing experimental autoimmune encephalomyelitis. CpG activation enhances the selective migration of pDC precursors to the inflamed spinal cord, induces their immediate production of TGF-β, and after migration, of enhanced levels of IL-27. CpG-pre-pDC derived TGF-β and IL-27 ensure protection at early and late phases of the disease, respectively. Spinal cords of CpG-pre-pDC-protected recipient mice display enhanced percentages of host-derived pDCs expressing TGF-β as well as an accumulation of IL-10 producing B cells and of CD11c

Topics: Animals; Bone Marrow Cells; Dendritic Cells; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Interleukin-27; Mice; Mice, Inbred C57BL; Mice, Knockout; Multiple Sclerosis; Spinal Cord; Toll-Like Receptor 9; Transforming Growth Factor beta

The Th17 immune response plays a key role in autoimmune diseases such as multiple sclerosis (MS) and inflammatory bowel disease (IBD). Expression of Th17-related genes in inflamed tissues has been reported in autoimmune diseases. However, values are frequently obtained using invasive methods. We aimed to identify biomarkers of MS in an accessible sample, such as blood, by quantifying the relative expression of 91 Th17-related genes in CD4+ T lymphocytes from patients with MS during a relapse or during a remitting phase. We also compared our findings with those of healthy controls. After confirmation in a validation cohort, expression of

Topics: Biomarkers; CD4-Positive T-Lymphocytes; Crohn Disease; Humans; Multiple Sclerosis; Signal Transduction; Smad2 Protein; Smad3 Protein; Smad4 Protein; Smad7 Protein; Sphingosine-1-Phosphate Receptors; Th17 Cells; Transforming Growth Factor beta

Diet has been suggested to be a potential environmental risk factor for the increasing incidence of autoimmune diseases, yet the underlying mechanisms remain elusive. Here, we show that high glucose intake exacerbated autoimmunity in mouse models of colitis and experimental autoimmune encephalomyelitis (EAE). We elucidated that high amounts of glucose specifically promoted T helper-17 (Th17) cell differentiation by activating transforming growth factor-β (TGF-β) from its latent form through upregulation of reactive oxygen species (ROS) in T cells. We further determined that mitochondrial ROS (mtROS) are key for high glucose-induced TGF-β activation and Th17 cell generation. We have thus revealed a previously unrecognized mechanism underlying the adverse effects of high glucose intake in the pathogenesis of autoimmunity and inflammation.

Topics: Animals; Autoimmunity; Cell Differentiation; Cells, Cultured; Diet; Disease Models, Animal; Eating; Encephalomyelitis, Autoimmune, Experimental; Glucose; Humans; Inflammation; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mitochondria; Multiple Sclerosis; Reactive Oxygen Species; Th17 Cells; Transforming Growth Factor beta

Environmental triggers acting at the intestinal barrier are thought to contribute to the initiation of autoimmune disorders. The transforming growth factor beta inhibitor Smad7 determines the phenotype of CD4

Topics: Animals; Autoimmunity; CD4-Positive T-Lymphocytes; Cell Differentiation; Central Nervous System; Disease Models, Animal; Encephalomyelitis; Encephalomyelitis, Autoimmune, Experimental; Gastrointestinal Microbiome; Gene Expression Regulation; Humans; Immune Tolerance; Inflammation; Intestines; Mice; Mice, Transgenic; Multiple Sclerosis; Signal Transduction; Smad7 Protein; Spinal Cord; Transforming Growth Factor beta

Myeloid-derived suppressor cells (MDSCs) are a diverse group of cells that are recognized for their remarkable suppressive effects on pro-inflammatory T cells. The pleiotropic nature of these cells, however, has been demonstrated by their differential effects on immune responses in different settings. Our and others' work has demonstrated suppressive effects of these cells. We previously demonstrated that these cells were mobilized to the lungs during experimental autoimmune encephalomyelitis (EAE), which is a murine model of multiple sclerosis, and potently inhibited CD8

Topics: Animals; Autoimmunity; Cell Communication; Cell Lineage; Cell Movement; Central Nervous System; Coculture Techniques; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Gene Expression; Humans; Interleukin-17; Lung; Mice, Inbred C57BL; Multiple Sclerosis; Myelin-Oligodendrocyte Glycoprotein; Myeloid-Derived Suppressor Cells; Peptide Fragments; Pertussis Toxin; Signal Transduction; Th17 Cells; Transforming Growth Factor beta

Current strategies for treating autoimmunity involve the administration of broad-acting immunosuppressive agents that impair healthy immunity. Intravenous (i.v.) administration of poly(lactide- co-glycolide) nanoparticles (NPs) containing disease-relevant antigens (Ag-NPs) have demonstrated antigen (Ag)-specific immune tolerance in models of autoimmunity. However, subcutaneous (s.c.) delivery of Ag-NPs has not been effective. This investigation tested the hypothesis that codelivery of the immunomodulatory cytokine, transforming growth factor beta 1 (TGF-β), on Ag-NPs would modulate the immune response to Ag-NPs and improve the efficiency of tolerance induction. TGF-β was coupled to the surface of Ag-NPs such that the loadings of Ag and TGF-β were independently tunable. The particles demonstrated bioactive delivery of Ag and TGF-β in vitro by reducing the inflammatory phenotype of bone marrow-derived dendritic cells and inducing regulatory T cells in a coculture system. Using an in vivo mouse model for multiple sclerosis, experimental autoimmune encephalomyelitis, TGF-β codelivery on Ag-NPs resulted in improved efficacy at lower doses by i.v. administration and significantly reduced disease severity by s.c. administration. This study demonstrates that the codelivery of immunomodulatory cytokines on Ag-NPs may enhance the efficacy of Ag-specific tolerance therapies by programming Ag presenting cells for more efficient tolerance induction.

Topics: Animals; Antigens; Cells, Cultured; Encephalomyelitis, Autoimmune, Experimental; Female; Immune Tolerance; Immunologic Factors; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Nanoconjugates; Polyglactin 910; Transforming Growth Factor beta

Topics: Animals; Autoimmunity; Cell Differentiation; Encephalomyelitis, Autoimmune, Experimental; Female; Gene Expression; Humans; Mice; Mice, Inbred C57BL; MicroRNAs; Multiple Sclerosis; Signal Transduction; Smad2 Protein; Smad3 Protein; Th17 Cells; Transforming Growth Factor beta

Extracellular matrix (ECM) deposition in active demyelinating multiple sclerosis (MS) lesions may impede axonal regeneration and can modify immune reactions. Response gene to complement (RGC)-32 plays an important role in the mediation of TGF-β downstream effects, but its role in gliosis has not been investigated. To gain more insight into the role played by RGC-32 in gliosis, we investigated its involvement in TGF-β-induced ECM expression and the upregulation of the reactive astrocyte markers α-smooth muscle actin (α-SMA) and nestin. In cultured neonatal rat astrocytes, collagens I, IV, and V, fibronectin, α-SMA, and nestin were significantly induced by TGF-β stimulation, and RGC-32 silencing resulted in a significant reduction in their expression. Using astrocytes isolated from RGC-32 knock-out (KO) mice, we found that the expression of TGF-β-induced collagens I, IV, and V, fibronectin, and α-SMA was significantly reduced in RGC-32 KO mice when compared with wild-type (WT) mice. SIS3 inhibition of Smad3 phosphorylation was also associated with a significant reduction in RGC-32 nuclear translocation and TGF-β-induced collagen I expression. In addition, during experimental autoimmune encephalomyelitis (EAE), RGC-32 KO mouse astrocytes displayed an elongated, bipolar phenotype, resembling immature astrocytes and glial progenitors whereas those from WT mice had a reactive, hypertrophied phenotype. Taken together, our data demonstrate that RGC-32 plays an important role in mediating TGF-β-induced reactive astrogliosis in EAE. Therefore, RGC-32 may represent a new target for therapeutic intervention in MS.

Topics: Actins; Animals; Astrocytes; Cells, Cultured; Collagen; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Extracellular Matrix; Female; Fibril-Associated Collagens; Gliosis; Humans; Mice; Mice, Knockout; Multiple Sclerosis; Nestin; Nuclear Proteins; Rats; RNA, Small Interfering; Transforming Growth Factor beta

Multiple sclerosis is a neurodegenerative disease characterized by the present of leukocytes in the brain tissue and subsequently the formation of sclerotic plaques. Leukocytes penetration into the blood-brain barrier is related to several factors, such as, the conversion of leukocyte gene expression or plasma characteristics. In this frame, we explore alteration of matrix metalloproteinase-2 (MMP-2), transforming growth factor beta (TGF-β) family, and Claudin-11 (as a main myelin structural protein) in leukocytes and blood plasma of multiple sclerosis patients compared to the normal group. Blood samples were collected from thirteen men affected by MS and fifteen healthy men. Leukocyte gene expression was measured using real-time PCR and plasma parameters were examined by ELISA. The results of this study showed that the gene expression of Claudin-11 was significantly higher in MS group compared with normal. Interestingly, the MMP-2 pattern was similar to Claudin-11 and correlated positively with it. It was observed that, although the expressions of TGF-β1 and TGF-β2 are down-regulated in the leukocytes of subjects with MS, they showed higher levels of these cytokines in blood plasma. The plasma level of TGF-β3 in MS patients was higher than normal and correlated with Claudin-11 concentration. In conclusion, the aberrant pattern of Claudin-11, TGF-βs family, and MMP-2 expression in leukocytes of the MS patients was observed in this study. Moreover, the plasma levels of TGF-βs family increased in the MS group. The findings of this study provide clues for further investigations to assay MS pathogenesis.

Topics: Adult; Blood-Brain Barrier; Claudins; Female; Gene Expression; Humans; Leukocytes; Male; Matrix Metalloproteinase 2; Middle Aged; Multiple Sclerosis; Transforming Growth Factor beta; Young Adult

Multifocal inflammatory lesions featuring destruction of lipid-rich myelin are pathologic hallmarks of multiple sclerosis. Lesion activity is assessed by the extent and composition of myelin uptake by myeloid cells present in such lesions. In the inflamed CNS, myeloid cells are comprised of brain-resident microglia, an endogenous cell population, and monocyte-derived macrophages, which infiltrate from the systemic compartment. Using microglia isolated from the adult human brain, we demonstrate that myelin phagocytosis is dependent on the polarization state of the cells. Myelin ingestion is significantly enhanced in cells exposed to TGF-β compared with resting basal conditions and markedly reduced in classically activated polarized cells. Transcriptional analysis indicated that TGF-β-treated microglia closely resembled M0 cells. The tyrosine kinase phagocytic receptor MerTK was one of the most upregulated among a select number of differentially expressed genes in TGF-β-treated microglia. In contrast, MerTK and its known ligands, growth arrest-specific 6 and Protein S, were downregulated in classically activated cells. MerTK expression and myelin phagocytosis were higher in CNS-derived microglia than observed in monocyte-derived macrophages, both basally and under all tested polarization conditions. Specific MerTK inhibitors reduced myelin phagocytosis and the resultant anti-inflammatory biased cytokine responses for both cell types. Defining and modulating the mechanisms that regulate myelin phagocytosis has the potential to impact lesion and disease evolution in multiple sclerosis. Relevant effects would include enhancing myelin clearance, increasing anti-inflammatory molecule production by myeloid cells, and thereby permitting subsequent tissue repair.

Topics: Adult; Brain; c-Mer Tyrosine Kinase; Cell Polarity; Cells, Cultured; Down-Regulation; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Macrophages; Microglia; Multiple Sclerosis; Myelin Sheath; Myeloid Cells; Phagocytosis; Protein S; Proto-Oncogene Proteins; Receptor Protein-Tyrosine Kinases; Transforming Growth Factor beta; Up-Regulation

Fingolimod, an oral therapeutic agent approved for patients with relapsing-remitting Multiple Sclerosis (MS), has been shown to prevent lymphocyte egress from secondary lymphoid tissues; however the specific drug effect on B cells in fingolimod-treated patients remains to be fully elucidated. We present here a comprehensive analysis on the proportions of B cell subsets in the periphery, and the levels of activation, functional surface markers and cytokine profile of B cells in MS patients, following initiation of fingolimod therapy, using flow cytometry and cytokine bead array. Fingolimod therapy increased the ratio of naïve to memory cells, elevated the percentage of plasma cells and highly increased the proportion of transitional B cells as well as additional regulatory subsets, including: IL10(+), CD25(+) and CD5(+) B cells. The percentage of activated CD69(+) cells was highly elevated in the remaining circulating B cells, which produced increased levels of IL10, TGFβ, IL6, IL4, LTα, TNFα and IFNγ cytokines, with an overall increased ratio of TGFβ to pro-inflammatory cytokines. Furthermore, fingolimod therapy reduced ICAM-1(+) cells, suggesting a possible reduction in antigen-presenting capacity. Phosphorylated-fingolimod was shown in vitro to reduce S1PR1 RNA and protein, to slightly increase viability and to activate anti-apoptotic Bcl2 in transformed B cells of patients with MS. In conclusion, fingolimod therapy modulates significantly the composition of circulating B cells, promoting regulatory subsets and an anti-inflammatory cytokine repertoire.

Topics: Adult; Antigen Presentation; B-Lymphocyte Subsets; Cell Differentiation; Cell Survival; Cytokines; Female; Fingolimod Hydrochloride; Gene Expression; Humans; Immunologic Memory; Immunophenotyping; Immunosuppressive Agents; Inflammation Mediators; Interleukin-10; Leukocyte Count; Lymphocyte Activation; Lymphocyte Count; Male; Middle Aged; Multiple Sclerosis; Plasma Cells; Receptors, Lysosphingolipid; Sphingosine-1-Phosphate Receptors; Transforming Growth Factor beta; Young Adult

Transforming growth factor beta (TGFβ) signalling is critical for regulatory T cell development and function, and regulatory T cell dysregulation is a common observation in autoimmune diseases, including multiple sclerosis. In a comprehensive miRNA profiling study of patients with multiple sclerosis naïve CD4 T cells, 19 differentially expressed miRNAs predicted to target the TGFβ signalling pathway were identified, leading to the hypothesis that miRNAs may be responsible for the regulatory T cell defect observed in patients with multiple sclerosis. Patients with multiple sclerosis had reduced levels of TGFβ signalling components in their naïve CD4 T cells. The differentially expressed miRNAs negatively regulated the TGFβ pathway, resulting in a reduced capacity of naïve CD4 T cells to differentiate into regulatory T cells. Interestingly, the limited number of regulatory T cells, that did develop when these TGFβ-targeting miRNAs were overexpressed, were capable of suppressing effector T cells. As it has previously been demonstrated that compromising TGFβ signalling results in a reduced regulatory T cell repertoire insufficient to control autoimmunity, and patients with multiple sclerosis have a reduced regulatory T cell repertoire, these data indicate that the elevated expression of multiple TGFβ-targeting miRNAs in naïve CD4 T cells of patients with multiple sclerosis impairs TGFβ signalling, and dampens regulatory T cell development, thereby enhancing susceptibility to developing multiple sclerosis.

Topics: Animals; CD4-Positive T-Lymphocytes; Cell Differentiation; Gene Expression; Humans; Mice; MicroRNAs; Multiple Sclerosis; Signal Transduction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

B lymphocytes contribute to the pathogenesis of Multiple Sclerosis (MS) by secreting antibodies and producing cytokines. This latter function was analyzed in myelin olygodendrocyte protein (MOG)-stimulated CD19+ B lymphocytes of 71 MS patients with different disease phenotypes and 40 age-and sex-matched healthy controls (HC). Results showed that: 1) CD19+/TNFα+, CD19+/IL-12+ and CD19+/IFNγ+ lymphocytes are significantly increased in primary progressive (PP) compared to secondary progressive (SP), relapsing-remitting (RR), benign (BE) MS and HC; 2) CD19+/IL-6+ lymphocytes are significantly increased in PP, SP and RR compared to BEMS and HC; and 3) CD19+/IL-13+, CD19+/IL-10+, and CD19+/IL-10+/TGFβ+ (Bregs) B lymphocytes are reduced overall in MS patients compared to HC. B cells expressing BTLA, a receptor whose binding to HVEM inhibits TcR-initiated cytokine production, as well as CD19+/BTLA+/IL-10+ cells were also significantly overall reduced in MS patients compared to HC. Analyses performed in RRMS showed that fingolimod-induced disease remission is associated with a significant increase in Bregs, CD19+/BTLA+, and CD19+/BTLA+/IL-10+ B lymphocytes. B lymphocytes participate to the pathogenesis of MS via the secretion of functionally-diverse cytokines that might play a role in determining disease phenotypes. The impairment of Bregs and CD19+/BTLA+ cells, in particular, could play an important pathogenic role in MS.

Topics: Adult; Antigens, CD19; B-Lymphocytes, Regulatory; Cells, Cultured; Female; Fingolimod Hydrochloride; Humans; Immunosuppressive Agents; Interferon-gamma; Interleukin-10; Interleukin-12; Male; Middle Aged; Multiple Sclerosis; Receptors, Immunologic; Severity of Illness Index; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Young Adult

Upregulation and/or maintenance of regulatory T cells (Tregs) during autoimmune insults may have therapeutic efficacy in autoimmune diseases. Earlier we have reported that sodium benzoate (NaB), a metabolite of cinnamon and a Food and Drug Administration-approved drug against urea cycle disorders, upregulates Tregs and protects mice from experimental allergic encephalomyelitis, an animal model of multiple sclerosis. However, mechanisms by which NaB increases Tregs are poorly understood. Because TGF-β is an important inducer of Tregs, we examined the effect of NaB on the status of TGF-β. In this study, we demonstrated that NaB induced the expression of TGF-β mRNA and protein in normal as well as proteolipid protein-primed splenocytes. The presence of a consensus STAT6 binding site in the promoter of the TGF-β gene, activation of STAT6 in splenocytes by NaB, recruitment of STAT6 to the TGF-β promoter by NaB, and abrogation of NaB-induced expression of TGF-β in splenocytes by small interfering RNA knockdown of STAT6 suggest that NaB induces the expression of TGF-β via activation of STAT6. Furthermore, we demonstrated that blocking of TGF-β by neutralizing Abs abrogated NaB-mediated protection of Tregs and experimental allergic encephalomyelitis. These studies identify a new function of NaB in upregulating TGF-β via activation of STAT6, which may be beneficial in MS patients.

Topics: Animals; Antibodies, Blocking; Cells, Cultured; Cinnamomum zeylanicum; Encephalomyelitis, Autoimmune, Experimental; Female; Food Preservatives; Forkhead Transcription Factors; Humans; Mice; Mice, Inbred C57BL; Mice, Transgenic; Multiple Sclerosis; Myelin Proteolipid Protein; Peptide Fragments; Promoter Regions, Genetic; Sodium Benzoate; STAT6 Transcription Factor; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Up-Regulation

miRNAs are key regulators in multiple sclerosis. To gain a better understanding of the molecular mechanisms of multiple sclerosis, differentially expressed microRNAs (DE-miRNAs) and genes (DEGs) were analyzed.. The miRNA expression profile GSE43590 including 11 samples of peripheral blood T-cells from relapsing-remitting MS patients and 9 normal samples as well as gene expression profile GSE52139 including 8 periplaque samples and 8 normal samples were downloaded from Gene Expression Omnibus. Then, DE-miRNAs and DEGs were identified using limma. Moreover, the target genes of DE-miRNAs were screened. Additionally, the integrated regulatory network of DEGs, DE-miRNAs and targets was constructed using Cytoscape. What's more, the functional modules were also screened using MINE in Cytoscape. Lastly, the functional annotation of genes in modules was conducted using DAVID.. A total of 2394 DEGs were screened in 8 periplaque samples. Additionally, 296 DE-miRNAs were identified in the 11 samples of peripheral blood T-cells from relapsing-remitting MS patients. Besides, 6 functional modules (A-F) were screened. Among them, has-miR-197 could target HNF4A. What's more, HNF4A could interact with CYP3A4. Additionally, has-miR-125b could target ID1 and ID3. Besides, ID1 could interact with THBS1. Furthermore, functional enrichment showed that CYP3A4 was significantly related to vitamin metabolic process. For the pathway enrichment, ID1 and ID3 were significantly enriched in TGF-beta signaling pathway.. Some important DE-miRNAs (such as has-miR-197and has-miR-125b) might be crucial for MS by regulating the expressions of their target genes.

Topics: Adult; Female; Gene Regulatory Networks; Genetic Markers; Humans; Male; Microarray Analysis; MicroRNAs; Middle Aged; Multiple Sclerosis; RNA, Untranslated; Transcriptome; Transforming Growth Factor beta

Experimental autoimmune encephalomyelitis (EAE) is a CD4(+)-mediated autoimmune pathology of the central nervous system (CNS) that is used as a model for the study of the human neuroinflammatory disease, multiple sclerosis. During the development of EAE, auto-reactive Th1 and Th17 CD4(+) T cells infiltrate the CNS promoting inflammatory cells recruitment, focal inflammation and tissue destruction. In this sense, statins, agents used to lower lipid levels, have recently shown to exert interesting immunomodulatory function. In fact, statins promote a bias towards a Th2 response, which ameliorates the clinical outcome of EAE. Additionally, simvastatin can inhibit Th17 differentiation. However, many other effects exerted on the immune system by statins have yet to be clarified, in particular during neuroinflammation. Thus, the aim of this study was to investigate the effects of simvastatin on the development of experimental autoimmune encephalomyelitis.. Mice were immunized with MOG(35-55) and EAE severity was assessed daily and scored using a clinical scale. Cytokine secretion by mononuclear cells infiltrating the CNS was evaluated by flow cytometry.. Simvastatin (5 mg/kg/day) improved clinical outcome, induced an increase in TGF-β mRNA expression and inhibited IL-6, IL-12p40, IL-12p70, RANTES and MIP-1β secretion (p < 0.05). This was accompanied by a significant decrease in CNS inflammatory mononuclear cell infiltration, with reduced frequencies of both Th1 and Th17 cells. Simvastatin inhibited the proliferation of T lymphocytes co-cultured with primary microglial cells.. Simvastatin treatment promotes EAE clinical amelioration by inhibiting T cell proliferation and CNS infiltration by pathogenic Th1 and Th17 cells.

Topics: Animals; Cell Differentiation; Cell Proliferation; Central Nervous System; Chemokine CCL5; Encephalomyelitis, Autoimmune, Experimental; Female; Inflammation; Interleukin-12 Subunit p40; Interleukin-6; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Simvastatin; Th1 Cells; Th17 Cells; Transforming Growth Factor beta

Accumulative evidence demonstrates that multiple sclerosis (MS) is caused by activation of myelin Ag-reactive CD4+ T cells. Therefore, the CD4+ T cells specific for myelin Ag may be the important therapeutical target of MS. The novel coinhibitory receptor B and T lymphocyte attenuator (BTLA) may have a regulatory role in maintaining peripheral tolerance, however, its role in MS is still unknown. In this study, a novel nanoparticle containing MOG peptide with BTLA was designed and transduced into dendritic cells (DCs), and MOG peptide-induced EAE mice were administrated with the genetically modified DCs in vivo. The results demonstrated that modified DCs significantly enhanced the proportion of Foxp3+ CD4+ regulatory T cells, increased IL-10 and TGF-β cytokine secretion, while decreased IL-2 and IFN-γ cytokine secretion. Furthermore, modified DCs supressed the CD4+ T cell response to MOG, cell infiltration into spinal cord, and the severity of EAE. In contrast, immune response to irrelevant exogenous Ag was not impaired by treatment with modified DCs. These findings suggested that DCs transduced with nanoparticle could induce specific CD4+ T-cells tolerance, which provided a promising therapeutic means to negatively manipulate immune response for autoimmune diseases without inhibition of the immune response to irrelevant Ag.

Topics: Animals; CD4-Positive T-Lymphocytes; Dendritic Cells; Encephalomyelitis, Autoimmune, Experimental; Female; Forkhead Transcription Factors; Gene Transfer Techniques; Immune Tolerance; Interferon-gamma; Interleukin-10; Interleukin-2; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Myelin Sheath; Myelin-Oligodendrocyte Glycoprotein; Nanoparticles; Receptors, Immunologic; Transforming Growth Factor beta

Multiple sclerosis (MS) is a presumed autoimmune disease directed against central nervous system (CNS) myelin, in which diet and obesity are implicated as risk factors. Immune responses can be influenced by molecules produced by fat cells, called adipokines. Adiponectin is an adipokine with anti-inflammatory effects. We tested the hypothesis that adiponectin has a protective role in the EAE model for MS, that can be induced by immunization with myelin antigens or transfer of myelin-specific T lymphocytes. Adiponectin deficient (ADPKO) mice developed worse EAE with greater CNS inflammation, demyelination, and axon injury. Lymphocytes from myelin-immunized ADPKO mice proliferated more, produced higher amounts of IFN-γ, IL-17, TNF-α, IL-6, and transferred more severe EAE than wild type (WT) lymphocytes. At EAE peak, the spleen and CNS of ADPKO had fewer regulatory T (Treg) cells than WT mice and during EAE recovery, Foxp3, IL-10 and TGF-β expression levels in the CNS were reduced in ADPKO compared with WT mice. Treatment with globular adiponectin in vivo ameliorated EAE, and was associated with an increase in Treg cells. These data indicate that adiponectin is an important regulator of T-cell functions during EAE, suggesting a new avenue of investigation for MS treatment.

Topics: Adiponectin; Adoptive Transfer; Animals; Autoimmunity; Cell Proliferation; Cells, Cultured; Central Nervous System; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Forkhead Transcription Factors; Interferon-gamma; Interleukin-10; Interleukin-17; Interleukin-6; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Knockout; Multiple Sclerosis; Myelin Sheath; Risk Factors; Spleen; T-Lymphocytes, Regulatory; Th1 Cells; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Mesenchymal stem cells (MSC) are currently strong candidates for stem cell therapy. Cytokines have a profound effect on the resultant immune responses. This study aims to evaluate variations in the cytokine profile of multiple sclerosis patients treated with autologous MSC.. Twenty five patients received one dose of intrathecal MSCs (mean number: 29.5 × 10⁶). To measure the gene expression of FOXP3, IFN-γ, TGF-β, IL-4, IL-10, IL-6, and their serum proteins, samples were collected at five intervals: day 0 prior to injection and months 1, 3, 6, and 12 after MSC therapy. Gene expression was evaluated via real-time PCR and protein values were measured by ELISA.. There were no statistically significant variations in gene expression and serum level of cytokines after a 1-year follow-up of MSC-treated MS patients. The only correlation found was an increase in IL-6 gene expression in patients with progressive disease.. Intrathecal injection of MSCs does not affect cytokine variation in peripheral blood. Because the condition of most of our patients either improved or stabilized after stem cell therapy (SCT), we speculate that the immunomodulatory or neuroregenerative effects of MSC are exerted locally in the central nervous system.

Topics: Adult; Cytokines; Female; Follow-Up Studies; Humans; Interferon-gamma; Interleukin-10; Interleukin-4; Interleukin-6; Male; Mesenchymal Stem Cell Transplantation; Middle Aged; Multiple Sclerosis; Real-Time Polymerase Chain Reaction; Transforming Growth Factor beta; Young Adult

Maintenance of FOXP3 protein expression is crucial for differentiation and maturation of regulatory T (Treg) cells, which play important roles in immune homeostasis and immune tolerance. We demonstrate here that PDCD5 interacts with FOXP3, increases acetylation of FOXP3 in synergy with Tip60 and enhances the repressive function of FOXP3. In PDCD5 transgenic (PDCD5tg) mice, overexpression of PDCD5 enhanced the level of FOXP3 protein and percentage of CD4(+)CD25(+)FOXP3(+) cells. Naïve CD4(+) T cells from PDCD5tg mice were more sensitive to TGF-β-induced Treg polarization and expansion. These induced Tregs retained normal suppressive function in vitro. Severity of experimentally-induced autoimmune encephalomyelitis (EAE) in PDCD5tg mice was significantly reduced relative to that of wild-type mice. The beneficial effect of PDCD5 likely resulted from increases of Treg cell frequency, accompanied by a reduction of the predominant pathogenic Th17/Th1 response. Activation-induced cell death enhanced by PDCD5 was also linked to this process. This is the first report revealing that PDCD5 activity in T cells suppresses autoimmunity by modulating Tregs. This study suggests that PDCD5 serves as a guardian of immunological functions and that the PDCD5-FOXP3-Treg axis may be a therapeutic target for autoimmunity.

Topics: Acetylation; Animals; Apoptosis; Apoptosis Regulatory Proteins; Autoimmunity; CD4 Antigens; Cell Differentiation; Cell Line; Cell Proliferation; Encephalomyelitis, Autoimmune, Experimental; Female; Forkhead Transcription Factors; HEK293 Cells; Histone Acetyltransferases; Humans; Interleukin-2 Receptor alpha Subunit; Lymphocyte Activation; Lymphocyte Count; Lysine Acetyltransferase 5; Mice; Mice, Inbred C57BL; Mice, Transgenic; Multiple Sclerosis; Neoplasm Proteins; RNA Interference; RNA, Small Interfering; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Trans-Activators; Transforming Growth Factor beta

The regulatory effect of Liuwei Dihuang Pills (LDP) was studied on cytokines in mice with experimental autoimmune encephalomyelitis (EAE), a model for human multiple sclerosis (MS), induced by immunization with MOG(35-55) and complete Freund's adjuvant (CFA) supplemented with pertussis toxin (PTX). LDP was administrated orally for 40 days, and prednisone acetate (PA) was used as a control. The pathological changes in the spinal cords of mice were observed by light microscope with hematoxylin-eosin (HE) staining and transmission electron microscope (TEM). The protein and mRNA expression of tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β) in the spinal cords were assessed by immunohistochemistry and RT-PCR assay, and the cyclic adenosine monophosphate (cAMP) in mice plasma was measured by radioimmunoassay (RIA) on days 12, 25 and 40 post-immunization (PI). The results showed that inflammatory cells, demyelination and axonal loss were reduced, and that the protein and mRNA expression of TNF-α and the ratio of TNF-α/TGF-β were obviously decreased, to different extents. However, the levels of cAMP were enhanced in LDP-treated groups. These findings suggested that LDP regulates the cytokine balance in favor of T helper 1 (Th1)/regulatory T (Treg) cells, which depend on enhancement of cAMP levels. LDP has a potential role in the treatment of MS and other demyelinating diseases of the central nervous system.

Topics: Animals; Cytokines; Drugs, Chinese Herbal; Encephalomyelitis, Autoimmune, Experimental; Humans; Male; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Spinal Cord; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Human amniotic epithelial cells (hAEC) have stem cell-like features and immunomodulatory properties. Here we show that hAEC significantly suppressed splenocyte proliferation in vitro and potently attenuated a mouse model of multiple sclerosis (MS). Central nervous system (CNS) CD3(+) T cell and F4/80(+) monocyte/macrophage infiltration and demyelination were significantly reduced with hAEC treatment. Besides the known secretion of prostaglandin E2 (PGE2), we report the novel finding that hAEC utilize transforming growth factor-β (TGF-β) for immunosuppression. Neutralization of TGF-β or PGE2 in splenocyte proliferation assays significantly reduced hAEC-induced suppression. Splenocytes from hAEC-treated mice showed a Th2 cytokine shift with significantly elevated IL-5 production. While transferred CFSE-labeled hAEC could be detected in the lung, none were identified in the CNS or in lymphoid organs. This is the first report documenting the therapeutic effect of hAEC in a MS-like model and suggest that hAEC may have potential for use as therapy for MS.

Topics: Amnion; Animals; Cell Proliferation; Cells, Cultured; Cytokines; Dinoprostone; Disease Models, Animal; Epithelial Cells; Female; Humans; Immunosuppression Therapy; Interleukin-5; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Placenta; Pregnancy; Spinal Cord; Spleen; Th2 Cells; Transforming Growth Factor beta

Laquinimod is an orally administered drug under development for the treatment of Multiple Sclerosis (MS), lacking a fully elucidated mode of action. We assessed the immunomodulatory effects of laquinimod in vitro on human B cells from healthy or MS patients, cultured alone or with CD4(+) T cells. Laquinimod modulated B cell markers, mainly by increasing the regulatory ones CD25, IL10 and CD86, and decreased IL4, while increasing IL10 and TGFβ in both B and T cells, in a B cell-mediated manner. These findings shed additional light on the mechanisms underlying the effects of laquinimod in MS and potentially other immune-mediated diseases.

Topics: Adult; B-Lymphocytes; B7-2 Antigen; Cells, Cultured; Coculture Techniques; Female; Humans; Immunologic Factors; Interleukin-10; Interleukin-2 Receptor alpha Subunit; Interleukin-4; Male; Middle Aged; Multiple Sclerosis; Quinolones; T-Lymphocytes; Transforming Growth Factor beta; Young Adult

Transforming growth factor (TGF-β1) is a pleiotropic cytokine, secreted by immune and nonhematopoietic cells. TGF-β is involved in many different critical processes, such as embryonal development, cellular maturation and differentiation, wound healing, and immune regulation. It maintains immune homeostasis by acting as a potent immune suppressor through inhibition of proliferation, differentiation, activation, and effector function of immune cells. Paradoxically, depending on the context, it displays proinflammatory properties by being a potent chemoattractant for neutrophils and promoting inflammation. In addition, it does not only induce differentiation into the anti-inflammatory Treg cells, but also into the proinflammatory Th17 and Th9 cells and inhibits Th22 differentiation. TGF-β has been demonstrated to be involved in multiple pathologies. In infections, it protects against collateral damages caused by the immune system, but it also promotes immune evasion and chronic infections. In autoimmune diseases, a TGF-β dysfunction leads to the loss of tolerance to self-antigens. In cancer, TGF-β is a potent inhibitor of cell proliferation and acts as a tumor suppressor at the beginning of tumorogenesis. However, once the cells become resistant to TGF-β, it mainly supports tumor growth and metastasis by promoting immune evasion and angiogenesis. In asthma, it is assumed to promote allergen tolerance, but plays a detrimental role in irreversible remodeling of the airways. Despite the high numbers of TGF-β-targeted pathways, it is a promising drug target for treatment of autoimmunity, cancer, fibrosis, if cell specificity can be achieved.This review summarizes the progresses that have been accomplished on the understanding of TGF-β's signaling in the immune homeostasis and its role in pathogenesis.

Topics: Arthritis, Rheumatoid; Base Sequence; CD8 Antigens; Cells, Cultured; Cytokines; Diabetes Mellitus, Type 1; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Immune Tolerance; Interleukin-2; Interleukin-4; Malaria; Molecular Sequence Data; Multiple Sclerosis; Promoter Regions, Genetic; Signal Transduction; Transcription Factors; Transforming Growth Factor beta; Transforming Growth Factor beta1; Wound Healing

Th17 cells are a highly pro-inflammatory T-helper cell subset characterised by the expression of IL17. They have been implicated in a variety of allergic and autoimmune conditions. T-regulatory (Treg) cells, a subset of CD4 cells which express Foxp3, CD25, IL10 and TGFβ, can suppress the activity of Th17 cells. In this study, we show that the circulating levels of Th17 and Treg cells in peripheral blood are correlated; furthermore, the expression of the pro-inflammatory cytokine IL17 and the anti-inflammatory cytokine IL10 by CD4 cells are also correlated. However, we found no clear correlation between cerebrospinal fluid (CSF) IL10 and IL17 cytokine levels in MS, approaching a negative correlation at the time of relapse, and an overall negative correlation between CSF IL17 and TGFβ levels, suggesting a lack of central regulation of pro:anti-inflammatory balance in this demyelinating condition.

Topics: Cytokines; Enzyme-Linked Immunosorbent Assay; Humans; Inflammation; Interleukin-10; Interleukin-17; Multiple Sclerosis; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

Effector Th1 cells perpetuate inflammatory damage in a number of autoimmune diseases, including MS and its animal model EAE. Recently, a self-regulatory mechanism was described in which effector Th1 cells produce the immunomodulatory cytokine IL-10 to dampen the inflammatory response in both normal and autoimmune inflammation. While the presence of TGF-β has been suggested to enhance and stabilize an IFN-γ(+) IL-10(+) phenotype, the molecular mechanism is poorly understood. Additionally, in the context of adoptive transfer EAE, it is unclear whether IL-10 acts on the transferred Th1 cells or on endogenous host cells. In the present study, using myelin-specific TCR-Tg mice, we show that repetitive Ag stimulation of effector Th1 cells in the presence of TGF-β increases the population of IFN-γ(+) IL-10(+) cells, which correlates with a decrease in EAE severity. Additionally, TGF-β signaling causes binding of Smad4 to the IL-10 promoter, providing molecular evidence for TGF-β-mediated IL-10 production from Th1 effector cells. Finally, this study demonstrates that IL-10 not only reduces encephalitogenic markers such as IFN-γ and T-bet on Th1 effector cells expressing the IL-10R but also prevents recruitment of both transferred and host-derived inflammatory T cells. These data establish a regulatory mechanism by which highly activated Th1 effector cells modulate their pathogenicity through the induction of IL-10.

Topics: Adoptive Transfer; Animals; Cells, Cultured; Encephalomyelitis, Autoimmune, Experimental; Flow Cytometry; Forkhead Transcription Factors; Interferon-gamma; Interleukin-10; Interleukin-17; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Promoter Regions, Genetic; Receptors, Antigen, T-Cell; RNA Interference; RNA, Small Interfering; Signal Transduction; Smad4 Protein; T-Box Domain Proteins; Th1 Cells; Transforming Growth Factor beta

Dysfunction of adenosinergic systems has been implicated in the development of multiple sclerosis in humans and experimental autoimmune encephalomyelitis (EAE) in animals. Caffeine, a non-selective antagonist of adenosine receptors, has been shown to provide protection against myelin oligodendroglia glycoprotein (MOG)-induced EAE in mice. In this study, we showed that chronic caffeine similarly imparts neuroprotection against EAE induced in rats by guinea pig spinal cord homogenates (GPSCH). GPSCH-induced EAE is characterized by extensive tissue inflammation with a typical chronic disease course. We showed that caffeine decreases the incidence of EAE and attenuates EAE pathology at behavioral, histological (inflammatory cell infiltration and demyelination) and neurochemical (expression of inflammatory cytokines) levels. The attenuation of GPSCH-induced pathology by chronic caffeine treatment was observed at doses of 10 and 30 mg/kg and during both peak and recovery phases of EAE. Furthermore, it was showed that chronic treatment with caffeine up-regulated A1 receptor and TGF-beta mRNAs and suppressed interferon-gamma mRNA in EAE rats. Together with previous reports, our data demonstrates that chronic treatment with caffeine exerts a neuroprotective effect against EAE, possibly through an A(1) receptor-mediated shift from Th1 to Th2 cell function, and provides a neurobiological basis for epidemiological investigation into the possible relationship between caffeine consumption and development of multiple sclerosis in humans.

Topics: Animals; Caffeine; Central Nervous System; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Down-Regulation; Drug Administration Schedule; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Interferon-gamma; Multiple Sclerosis; Rats; Rats, Wistar; Receptor, Adenosine A1; Receptor, Adenosine A2A; RNA, Messenger; Spinal Cord; Subcellular Fractions; Th1 Cells; Th2 Cells; Transforming Growth Factor beta

Multiple sclerosis (MS) is a chronic inflammatory response against constituents of the central nervous system. It is known that regulatory T cells (Tregs) play a key role in the autoimmune balance and their improper function may facilitate the expansion of autoaggressive T cell clones. Recently, microRNAs (miRNAs) have been involved in autoimmune disorders and their loss-of-function in immune cells was shown to facilitate systemic autoimmune disorders. Here, we analyzed the miRNA expression profile in Tregs from MS-RR.. We assessed miRNA genome-wide expression profile by microarray analysis on CD4(+)CD25(+high) T cells from 12 MS relapsing-remitting patients in stable condition and 14 healthy controls. Since CD4(+)CD25(+high) T cells comprise both T regulatory cells (CD4(+)CD25(+high)CD127(dim/-)) and T effector cells (CD4(+)CD25(+high)CD127(+)), we performed a quantitative RT-PCR on CD4(+)CD25(+high)CD127(dim/-) and CD4(+)CD25(+high)CD127(+) cells isolated from the same blood sample.. We found 23 human miRNAs differentially expressed between CD4(+)CD25(high)bona fide Treg cells from MS patients vs. healthy donors, but, conversely, among the deregulated miRNAs, members of the miR-106b-25 were found down-regulated in MS patients when compared to healthy donors in CD4(+)CD25(high)CD127(dim/-) T regulatory cells. More interesting, the ratio between Treg/Teff showed an enrichment of these microRNA in T regulatory cells derived from patients if compared to healthy controls.. miR-106b and miR-25 were previously shown to modulate the TGF-β signaling pathway through their action on CDKN1A/p21 and BCL2L11/Bim. TGF-β is involved in T regulatory cells differentiation and maturation. Therefore, the deregulation of this miRNA cluster may alter Treg cells activity in course of MS, by altering TGF-β biological functions.

Topics: Adult; Analysis of Variance; Antigens, CD; Coculture Techniques; Female; Flow Cytometry; Gene Expression Profiling; Gene Expression Regulation; Genome-Wide Association Study; Humans; Lymphocyte Activation; Male; MicroRNAs; Middle Aged; Multiple Sclerosis; Oligonucleotide Array Sequence Analysis; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Experimental autoimmune encephalomylitis (EAE), an animal mode of multiple sclerosis (MS), was previously considered that be mediated by Th1 cells. However, a number of recent studies provided strong evidence that T helper cells that produce IL-17 play a dominant role in the pathogenesis of EAE. Curcumin (1,7-Bis 94-hydroxy-3-methoxyphenyl)-1,6 heptadiene-3, 5-di-one) is a naturally occurring polyphenolic phytochemical isolated from the rhizome of the medicinal plant Curcuma longa. It has been strongly implicated as an anti-inflammatory agent, but the precise mechanisms of its action are largely unknown. In the present study, we have investigated the efficacy and mechanism of curcumin against EAE. The treatment of Lewis rats with curcumin significantly reduced the clinical severity of EAE, and had a dramatic reduction in the number of inflammatory cells infiltration in the spinal cord. The proliferation of the MBP-reaction lymphocyte also was reduced in a curcumin dose-dependent manner. Furthermore, the mRNA expression of the cytokine profiles was assessed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR), revealing the dramatic decrease of IL-17, TGF-beta, IL-6, IL-21, STAT3, and RORgammat expression in curcumin-treated groups and STAT3-phosphorylation also was inhibited. These findings indicated that curcumin amelioration EAE was, to a large extent, due to inhibit differentiation and development of Th17 cells depends on down-regulating expression of IL-6, IL-21, RORgammat signaling and inhibition STAT3-phosphorylation, suggests it is useful in the treatment of MS and other Th17 cell-mediated inflammatory diseases.

Topics: Animals; Cell Proliferation; Curcuma; Curcumin; Disease Progression; Dose-Response Relationship, Drug; Down-Regulation; Encephalomyelitis, Autoimmune, Experimental; Female; Freund's Adjuvant; Guinea Pigs; Humans; Immunization; Interleukin-17; Interleukin-6; Interleukins; Jurkat Cells; Multiple Sclerosis; Myelin Basic Protein; Nuclear Receptor Subfamily 1, Group F, Member 3; Peptides; Phosphorylation; Phytotherapy; Rats; Rats, Inbred Lew; Rhizome; Spinal Cord; STAT3 Transcription Factor; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

Inhibition of remyelination is part of the complex problem of persistent dysfunction after spinal cord injury (SCI), and residual myelin debris may be a factor that inhibits remyelination. Phagocytosis by microglial cells and by macrophages that migrate from blood vessels plays a major role in the clearance of myelin debris. The object of this study was to investigate the mechanisms underlying the failure of significant remyelination after SCI.. The authors investigated macrophage recruitment and related factors in rats by comparing a contusion model (representing contusive SCI with residual myelin debris and failure of remyelination) with a model consisting of chemical demyelination by lysophosphatidylcholine (representing multiple sclerosis with early clearance of myelin debris and remyelination). The origin of infiltrating macrophages was investigated using mice transplanted with bone marrow cells from green fluorescent protein-transfected mice. The changes in levels of residual myelin debris and the infiltration of activated macrophages in demyelinated lesions were investigated by immunostaining at 2, 4, and 7 days postinjury. To investigate various factors that might be involved, the authors also investigated gene expression of macrophage chemotactic factors and adhesion factors.. Activated macrophages coexpressing green fluorescent protein constituted the major cell population in the lesions, indicating that the macrophages in both models were mainly derived from the bone marrow, and that very few were derived from the intrinsic microglia. Immunostaining showed that in the contusion model, myelin debris persisted for a long period, and the infiltration of macrophages was significantly delayed. Among the chemotactic factors, the levels of monocyte chemoattractant protein-1 and granulocyte-macrophage colony-stimulating factor were lower in the contusion model at 2 and 4 days postinjury.. The results suggest that the delayed infiltration of activated macrophages is related to persistence of myelin debris after contusive SCI, resulting in the inhibition of remyelination.

Topics: Animals; Bone Marrow Transplantation; Chemokine CCL2; Contusions; Cytokines; Demyelinating Diseases; Disease Models, Animal; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Intercellular Adhesion Molecule-1; Macrophage Activation; Macrophage Inflammatory Proteins; Macrophages; Mice; Mice, Inbred C57BL; Microglia; Multiple Sclerosis; Myelin Sheath; Phagocytosis; Platelet Endothelial Cell Adhesion Molecule-1; Rats; Rats, Sprague-Dawley; Spinal Cord Injuries; Transforming Growth Factor beta; Wound Healing

Experimental autoimmune encephalomyelitis (EAE) is a primary animal model of multiple sclerosis (MS). MS predominantly presents with evidence of lesions in the subcortical periventricular white matter regions of the brain. Research into the pathogenesis of the demyelinating lesions in the brain has been hampered by the fact that conventional models of EAE present with progressive ascending paralysis which recapitulates mainly the spinal cord lesions of multiple sclerosis. There is little evidence of brain involvement. Systemic administration of pertussis toxin (PTx) has been shown to induce the proinflammatory cascade of TGF-beta, IL-6, and Th17 in the central nervous system, which recently has been identified as essential in the development of EAE. To determine whether intracerebroventricular (icv) administration of PTx would result in subcortical periventricular demyelinating lesions in the brain, we examined the effect in a MOG induced EAE model. We found that icv PTx induced subcortical periventricular brain lesions that resemble the pathologic demyelinating lesions of MS. Moreover, icv PTx induced Th17 infiltration and increased expression of cytokines IL-6 and TGF-beta. We thus generated a highly reproducible model with remarkable histological similarities to the predominant demyelinating brain lesions seen in MS.

Topics: Animals; Cerebral Ventricles; Encephalomyelitis, Autoimmune, Experimental; Female; Interleukin-6; Leukocytes; Meningitis; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Pertussis Toxin; Transforming Growth Factor beta

To analyze the different responses of NK cell subsets isolated from the patients with proceeding multiple sclerosis to interferon-beta.. NK cells were isolated from the patients' peripheral blood mononuclear cells, separated into two subsets: CD94/NKG2A-bright and CD94/NKG2A-dim and then sorted by flow cytometry. The two subsets were cultured with IFN-beta to evaluate CD94/NKG2A expression pattern and the corresponding cell proliferation and secreted IL-10 and TGF-beta.. The positive NK cells of CD94/NKG2A made up 25.5% within the whole NK population. Among them, CD94/NKG2A-bright and CD94/NKG2A-dim amounted to 23.6% and 76.4%, respectively. When INF-beta was added to two groups, the proliferation rate of CD94/NKG2A-bright group was much lower than that of CD94/NKG2A-dim group and its expression pattern didn't alter greatly. In contrast, the expression of CD94/NKG2A in CD94/NKG2A-dim group was markedly increased. IL-10 and TGF-beta secretion was increased dominantly compared with the two untreated groups. There were great differences between the secreted IL-10 and TGF-beta in the stimulated groups.. IFN-beta can inhibit NK cells by inducing the inhibitory expression of CD94/NKG2A and stimulate IL-10 and TGF-beta secretion. CD94/NKG2A-bright group and CD94/NKG2A-dim group make different reaction to IFN-beta.

Topics: Cell Proliferation; Cells, Cultured; Flow Cytometry; Humans; Interferon-beta; Interleukin-10; Killer Cells, Natural; Multiple Sclerosis; NK Cell Lectin-Like Receptor Subfamily C; NK Cell Lectin-Like Receptor Subfamily D; Transforming Growth Factor beta

Systemic injection of small amounts of transforming growth factor-beta (TGF-beta), a cytokine produced by lymphoid and other cells, has a profound effect in protecting mice from the inflammatory demyelinating lesions of experimental allergic encephalomyelitis (EAE; an animal model for multiple sclerosis). However, TGF-beta has side-effects, which might be avoided if the cells producing TGF-beta can be delivered to the affected site in the nervous system to insure its local release in small amounts. Myelin basic protein (MBP)-specific, cloned CD4+ T cells were engineered by retroviral transduction to produce latent TGF-beta. Studies about the spontaneous form of EAE in T cell receptor (TCR)-transgenic recombination-activating gene (RAG)-1(-/-) mice showed that essentially all of the MBP-specific, TCR-transgenic RAG-1(-/-) (BALB/cxB10.PL)F1 mice develop spontaneous EAE by the age of 11 weeks. By 12 weeks, 25-50% of the mice have died from disease. A single injection of TGF-beta1-transduced T helper cell type 1 (Th1) cells significantly protected the mice from EAE, and untransduced Th1 cells did not protect. MBP-specific BALB/c Th2 clones, transduced with TGF-beta1-internal ribosome entry site-green fluorescent protein (GFP) significantly reduced EAE induction by untransduced Th1 cells in RAG-1(-/-) B10.PL mice. Furthermore, the GFP+ TGF-beta1-producing Th2 cells were detectable in the spinal cords of the injected mice.

Topics: Adoptive Transfer; Animals; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Homeodomain Proteins; Mice; Mice, Inbred BALB C; Mice, Knockout; Multiple Sclerosis; Myelin Basic Protein; Receptors, Antigen, T-Cell; Th1 Cells; Th2 Cells; Transduction, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1

Experimental autoimmune encephalomyelitis (EAE) is an instructive model for the human demyelinating disease multiple sclerosis. Lewis (LEW) rats immunized with myelin-basic protein (MBP) develop EAE characterized by a single episode of paralysis, from which they recover spontaneously and become refractory to a second induction of disease. LF 15-0195 is a novel molecule that has potent immunosuppressive effects in several immune-mediated pathological manifestations, including EAE. In the present study, we show that a 30-day course of LF 15-0195 treatment not only prevents MBP-immunized LEW rats from developing EAE but also preserves their refractory phase to reinduction of disease. This effect is Ag driven since it requires priming by the autoantigen during the drug administration. In contrast to other immunosuppressive drugs, short-term treatment with this drug induces a persistent tolerance with no rebound of EAE up to 4 mo after treatment withdrawal. This beneficial effect of LF 15-0195 on EAE does not result from the deletion of MBP-specific Vbeta8.2 encephalitogenic T cells. In contrast, this drug favors the differentiation of MBP-specific CD4 T cells into Foxp3-expressing regulatory T cells that, upon adoptive transfer in syngeneic recipients, prevent the development of actively induced EAE. Finally, we demonstrate that the tolerance induced by LF 15-0195 treatment is not dependent on the presence of TGF-beta. Together, these data demonstrate that short-term treatment with LF 15-0195 prevents MBP-immunized LEW rats from EAE by favoring the development of Foxp-3-expressing regulatory CD4 T cells.

Topics: Adoptive Transfer; Animals; Base Sequence; CD4-Positive T-Lymphocytes; DNA; Encephalomyelitis, Autoimmune, Experimental; Forkhead Transcription Factors; Gene Expression; Guanidines; Humans; Immunosuppressive Agents; Male; Multiple Sclerosis; Myelin Basic Protein; Neutralization Tests; Rats; Rats, Inbred Lew; Receptors, Antigen, T-Cell, alpha-beta; Receptors, Chemokine; Transforming Growth Factor beta

Topics: Animals; Cell Communication; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Humans; Mice; Multiple Sclerosis; Neurons; T-Lymphocytes; Transforming Growth Factor beta

In the central nervous system, basement membrane (BM) constituents are predominantly associated with the vasculature. However, under inflammatory conditions, the expression of BM components may alter. Here, we investigated the distribution of several BM components, including laminin, collagen type IV and heparan sulfate proteoglycans in various multiple sclerosis (MS) lesions. We observed irregular and discontinuous BMs in active lesions. Throughout active MS lesions, we found dense networks of BM proteins, which were surprisingly not associated with the cerebrovasculature. These striking parenchymal networks were not observed in chronic inactive MS lesions and brains of non-neurological controls. In addition, we studied the distribution of transforming growth factor-beta1 (TGF-beta1), since it is known as a major modulator of ECM production. Leukocytes, in particular CD68-positive macrophages, expressed high levels of TGF-beta1 and were located in close proximity to parenchymal BM deposits in the MS lesions. We postulate that these BM networks may play a role in the further recruitment of inflammatory cells and form a barrier for axonal regeneration.

Topics: Adult; Aged; Basement Membrane; Brain; Collagen Type IV; Female; Heparan Sulfate Proteoglycans; Humans; Immunohistochemistry; Laminin; Leukocytes; Male; Middle Aged; Multiple Sclerosis; Nerve Degeneration; Reference Values; Saccharomyces cerevisiae Proteins; Tissue Distribution; Transforming Growth Factor beta; Trehalase

Subjects from Sardinia, Italy, are relatively homogeneous compared to Swedes. Although ethnically distant, both populations have similarly high multiple sclerosis (MS) incidence rates. Pro- and anti-inflammatory cytokines and their receptors, signalling molecules and other immune response-associated factors might influence MS pathogenesis, though definite proof is missing. The study of populations with similar MS incidence but different genetic and environmental background could make possible the definition of factors that relate to such background differences. We selected untreated female MS patients from Sassari, Sardinia, and Stockholm, Sweden, and corresponding sex- and age-matched healthy controls (HC), to study blood mononuclear cells (MNC) for mRNA expression of 20 immune response-related genes considered relevant in MS, employing real-time PCR. Higher expression of IL-12p40 mRNA was confined to MS from both Sassari and Stockholm, compared to corresponding HC. MS patients from Sassari, but not Stockholm, expressed higher TNF-alpha compared to corresponding HC. MS patients from Stockholm, but not Sassari, expressed higher IL-6. Indoleamine 2,3 dioxygenase (IDO), a molecule necessary in tolerance induction, was lower in MS from Stockholm compared to corresponding HC. This was not observed in Sassari. No differences were detected for other members of the IL-12 family, other Th1 and Th2 cytokines, and the signalling molecules Stat 4 and 6. The results corroborate a pro-inflammatory state in MS as reflected by high expression of IL-12, TNF-alpha and IL-6, although the extent of expression of TNF-alpha, IL-6 and IDO differs between strictly matched MS patients from different high-incidence areas. This might result from genetic and/or environmental differences. They may account for some of the discrepancies regarding immune response-related molecules previously reported in MS. In conclusion, a pro-inflammatory state exists in MS patients from Sassari as well as Stockholm. The changes of pro-inflammatory and other immune response-related variables differ however between the two MS populations. This may be attributed to the genetic and/or environmental background.

Topics: Adult; Cytokines; DNA-Binding Proteins; Female; Genetic Heterogeneity; Humans; Indoleamine-Pyrrole 2,3,-Dioxygenase; Interleukin-10; Interleukin-12; Interleukin-18; Interleukin-18 Receptor alpha Subunit; Interleukin-4; Interleukin-6; Italy; Middle Aged; Multiple Sclerosis; Receptors, Cytokine; Receptors, Interleukin; Receptors, Interleukin-10; Receptors, Interleukin-12; Receptors, Interleukin-18; Receptors, Interleukin-6; Receptors, Tumor Necrosis Factor; STAT4 Transcription Factor; STAT6 Transcription Factor; Sweden; Trans-Activators; Transforming Growth Factor beta; Tryptophan Oxygenase; Tumor Necrosis Factor-alpha

Copolymer-I (COP-I) has unique immune regulatory properties and is a treatment option for multiple sclerosis (MS). This study revealed that COP-I induced the conversion of peripheral CD4+CD25- to CD4+CD25+ regulatory T cells through the activation of transcription factor Foxp3. COP-I treatment led to a significant increase in Foxp3 expression in CD4+ T cells in MS patients whose Foxp3 expression was reduced at baseline. CD4+CD25+ T cell lines generated by COP-I expressed high levels of Foxp3 that correlated with an increased regulatory potential. Furthermore, we demonstrated that the induction of Foxp3 in CD4+ T cells by COP-I was mediated through its ability to produce IFN-gamma and, to a lesser degree, TGF-beta1, as shown by antibody blocking and direct cytokine induction of Foxp3 expression in T cells. It was evident that in vitro treatment and administration with COP-I significantly raised the level of Foxp3 expression in CD4+ T cells and promoted conversion of CD4+CD25+ regulatory T cells in wild-type B6 mice but not in IFN-gamma knockout mice. This study provides evidence for the role and mechanism of action of COP-I in the induction of CD4+CD25+ regulatory T cells in general and its relevance to the treatment of MS.

Topics: Animals; CD4-Positive T-Lymphocytes; DNA Primers; DNA-Binding Proteins; Forkhead Transcription Factors; Gene Expression Regulation; Glatiramer Acetate; Humans; Immunotherapy; Interferon-gamma; Leukocytes, Mononuclear; Mice; Mice, Inbred C57BL; Mice, Knockout; Multiple Sclerosis; Peptides; Receptors, Interleukin-2; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta; Transforming Growth Factor beta1

Reliable, and easy to measure, immunological markers able to denote disease characteristics in multiple sclerosis (MS) patients are still lacking. We applied a multivariate statistical analysis on results obtained by measuring-by real-time RT-PCR-mRNA levels of 25 immunological relevant molecules in PBMCs from 198 MS patients. The combined measurement of mRNA levels of IL-1beta, TNF-alpha, TGF-beta, CCL20 and CCR3 was able to distinguish MS patients from healthy individuals. CXCR5, CCL5, and CCR3 combined mRNA levels identify primary progressive MS patients while TNF-alpha, IL-10, CXCL10 and CCR3 differentiate relapsing MS patients. Our results indicate that multi-parametric analysis of mRNA levels of immunological relevant molecules in PBMCs may represent a successful strategy for the identification of putative peripheral markers of disease state and disease activity in MS patients.

Topics: Adult; Biomarkers; Chemokine CCL20; Chemokine CCL5; Chemokines, CC; Disease Progression; Female; Humans; Interleukin-1; Linear Models; Macrophage Inflammatory Proteins; Magnetic Resonance Imaging; Male; Middle Aged; Models, Immunological; Multiple Sclerosis; Multiple Sclerosis, Chronic Progressive; Multivariate Analysis; Receptors, CCR3; Receptors, Chemokine; Receptors, CXCR5; Receptors, Cytokine; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Dysregulation in the expression of pro- and anti-inflammatory cytokines is one of the milestones in multiple sclerosis (MS) development and progression. The aim of this study was to investigate the possible influence of TNF-alpha (-308), TGF-beta (codons 10 and 25), IL-10 (-1082, -819, -592), IL-6 (-174) and IFN-gamma (+874) polymorphisms on susceptibility to multiple sclerosis (MS). Genotyping was performed by PCR-SSP method in 55 MS patients with relapsing-remitting form of the disease and 86 healthy subjects from Bulgarian population. We observed a statistically significant increase in the CC genotype of IL-10 -819 and -592 SNPs coupled with a decreased frequency of the TGF-beta +915 CG genotype in our MS patients (Pc<0.05). No significant differences were observed between MS patients and controls with respect to the distribution of the other cytokine gene polymorphisms investigated. Although the size of the study group is small, these results indicate that polymorphic variations of two of the major anti-inflammatory cytokines, IL-10 and TGF-beta, may play a role in MS susceptibility.

Topics: Adult; Animals; Bulgaria; Case-Control Studies; Chi-Square Distribution; Cytokines; Genetic Predisposition to Disease; Humans; Interleukin-10; Male; Multiple Sclerosis; Polymorphism, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta

Our objective was to investigate whether polymorphisms and haplotypes in the TGFB1 gene are associated with susceptibility or disease characteristics of multiple sclerosis (MS). In 247 MS patients and 194 controls, single nucleotide polymorphisms (SNPs) at position +869 (Leu10Pro) and position +915 (Arg25Pro) in the signaling sequence of the TGFB1 gene were determined, and the distribution of alleles, genotypes, and haplotypes was related to clinical data. In addition, magnetic resonance imaging (MRI) data were studied in a subgroup of patients (n = 96). The allele distribution of the two polymorphisms studied was in Hardy-Weinberg equilibrium in patients and in controls. No association was found with any of the three haplotypes found in the Dutch population, denoted as haplotype 1 (TGFB1+869T-TGFB1+915G), haplotype 2 (TGFB1+869C-TGFB1+915G), and haplotype 3 (TGFB1+869C-TGFB1+915C). However, the TGFB1+869 genotype CC was significantly more frequent in patients (p = 0.031, chi2 test). The highest frequency of the TGFB1+869 genotype CC was observed in male patients (25.2% vs. 10.0% in controls, p = 0.004, chi2 test), and carriership of TGFB1+869 allele C was correspondingly increased in male patients (74.8% vs. 56.7%, p = 0.008, chi2 test, OR 2.27, 95% CI 1.23-4.17). Although there was no association with clinical markers of disease progression, patients homozygous for TGFB1+869 allele C showed a significantly higher annual increase in two MRI parameters: ventricular fraction (central atrophy) and T1-hypointense lesion load (matrix destruction). The TGFB1 T+869C (Leu10Pro) gene polymorphism is associated with MS susceptibility, especially in males, and with a more destructive course of the disease as illustrated by MRI.

Topics: Adult; Base Sequence; Demography; Female; Genetic Predisposition to Disease; Humans; Magnetic Resonance Imaging; Male; Middle Aged; Molecular Sequence Data; Multiple Sclerosis; Polymorphism, Single Nucleotide; Sex Factors; Transforming Growth Factor beta; Transforming Growth Factor beta1

Monocyte infiltration of the brain is central to the pathogenesis of HIV-1 encephalitis. The cytokines promoting recruitment of monocytes into the central nervous system during HIV-1 infection are not established. In this study, we evaluated human cerebrospinal fluid from patients with HIV-1 infection for transforming growth factor beta1 (TGFbeta1) and monocyte chemotactic protein-1 (MCP-1) using a quantitative sandwich enzyme-linked immunoassays. Cytokine levels were compared to those from patients with multiple sclerosis and normal controls. In cerebrospinal fluid of patients with HIV-1 infection and CD4<500 cells/mm3, both TGFbeta1 and MCP-1 were significantly elevated compared to those with CD4>500 cells/mm3, multiple sclerosis, and controls.

Topics: CD4 Lymphocyte Count; Chemokine CCL2; HIV Infections; Humans; Immunocompromised Host; Multiple Sclerosis; Reference Values; Transforming Growth Factor beta

Nitric oxide (NO) has been implicated in the etiopathology of multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), and inhibition of NO synthesis has been proposed to be a possible mechanism of action of drugs to treat MS. In the present study, we investigated the inhibitory effect on NO synthesis of various steroids, cytokines and drugs used or proposed for the treatment of MS. As a model system, we used primary rat microglial cells which produce NO synthase and subsequently release NO upon stimulation with lipopolysaccharide (LPS). Among the substances tested, the glucocorticoids prednisone, hydrocortisone, dexamethasone and progesterone as well as transforming growth factor-beta (TGF-beta) dose-dependently inhibited LPS-induced nitric oxide synthase (iNOS) and NO synthesis. In contrast, COP-1, the phosphodiesterase inhibitors rolipram and pentoxifylline, the cytokines interleukin-10 (IL-10) and interferon-beta (IFN-beta) as well as the steroids beta-estradiol, testosterone, and dehydroepiandrosterone (DHEA) showed no inhibitory effect. Cholesterol slightly, but not significantly, increased LPS-induced nitric oxide synthesis. We conclude from the present study that with respect to treatment of MS, inhibition of NO synthesis may be an important mechanism of action of glucocorticoids and transforming growth factor-beta, but not of other drugs used or proposed to treat MS.

Topics: Algorithms; Animals; Anti-Inflammatory Agents; Blotting, Western; Cells, Cultured; Enzyme Induction; Interferon-gamma; Interleukin-1; Lipopolysaccharides; Microglia; Multiple Sclerosis; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Prednisolone; Rats; Transforming Growth Factor beta

Humoral auto-immunity to the myelin oligodendrocyte glycoprotein (MOG) is likely involved in the pathogenesis of multiple sclerosis (MS). In 44 MS patients and 30 controls, Ig-producing B cells were identified by their isotype and as MOG-specific spot-forming cells (SFC). Peripheral anti-MOG antibodies were assayed in ELISA as well as anti-butyrophilin antibodies to investigate for molecular mimicry. MS patients had significantly higher levels of IgA- and MOG-SFC than controls, as well as significantly higher antibody responses to MOG and butyrophilin. These data provide added support for the implication of anti-MOG humoral immunity in the pathophysiology of MS, and suggest a balance of systemic (anti-self) and mucosal (environment-modulated) immune reactions in an attempt at regulating the pathogenic specific immune response.

Topics: Adult; Aged; Aged, 80 and over; Antibody-Producing Cells; B-Lymphocytes; Butyrophilins; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Humans; Immunoglobulin Isotypes; Male; Membrane Glycoproteins; Middle Aged; Multiple Sclerosis; Myelin Proteins; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Peptide Fragments; Transforming Growth Factor beta

We investigated circulating anti-inflammatory and pro-inflammatory cytokines, and their ex vivo PBMC production in the absence or presence of the neuroantigens myelin basic protein (MBP) and myelin oligodendrocyte glycoprotein (MOG) and T cell mitogen (PHA) in MS patients in relapse and remission, patients with other neurological disorders (OND) and normal healthy controls. MS patients in relapse exhibited significantly increased PBMC production of TNF-alpha spontaneously compared with MS remission and healthy controls and with MBP compared with MS remission. Patients in relapse had significantly increased spontaneous, PHA- and MBP-induced PBMC IL-1beta production compared with remission MS, and was increased compared (PHA only) with OND and healthy controls. In relapse there was also significantly increased PBMC IFN-gamma production (PHA only) compared with remission and a significantly lower production of biologically active TGF-beta1 (PHA only) compared with remission MS and OND. In contrast, MS patients in remission produced significantly less spontaneous and MBP-induced TNF-alpha, spontaneous, PHA- and MBP-induced IL-1beta and PHA-induced IFN-gamma together with increased production of biologically active TGF-beta1. MOG non-specifically increased PBMC TNF-alpha and IL-1beta production in all groups. Pro-inflammatory cytokines in corresponding plasma samples were undetectable whilst the concentration of biologically active TGF-beta1 was the reverse of ex vivo PBMC findings. The increase in biologically active TGF-beta1 production ex vivo in OND patients, despite active disease, compared with the low level in the MS relapse may indicate a regulatory defect in MS. We conclude that the balance between biologically active TGF-beta1 and the pro-inflammatory TNF-alpha, IL-1beta and IFN-gamma is dysregulated during MS relapse-remission and that normal counter-regulatory mechanisms during the relapse phase are defective.

Topics: Adult; Cytokines; Female; Humans; Interferon-gamma; Leukocytes, Mononuclear; Male; Middle Aged; Multiple Sclerosis; Myelin Proteins; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Transforming Growth Factor beta

The ability of a remedy to modulate the pathological process in the target organ is crucial for its therapeutic activity. Glatiramer acetate (GA, Copaxone, Copolymer 1), a drug approved for the treatment of multiple sclerosis, induces regulatory T helper 2/3 cells that penetrate the CNS. Here we investigated whether these GA-specific T cells can function as suppressor cells with therapeutic potential in the target organ by in situ expression of T helper 2/3 cytokines and neurotrophic factors. GA-specific cells and their in situ expression were detected on the level of whole-brain tissue by using a two-stage double-labeling system: (i) labeling of the GA-specific T cells, followed by their adoptive transfer, and (ii) detection of the secreted factors in the brain by immunohistological methods. GA-specific T cells in the CNS demonstrated intense expression of the brain-derived neurotrophic factor and of two antiinflammatory cytokines, IL-10 and transforming growth factor beta. No expression of the inflammatory cytokine IFN-gamma was observed. This pattern of expression was manifested in brains of normal and experimental autoimmune encephalomyelitis-induced mice to which GA-specific cells were adoptively transferred, but not in control mice. Furthermore, infiltration of GA-induced cells to the brain resulted in bystander expression of IL-10 and transforming growth factor beta by resident astrocytes and microglia. The ability of infiltrating GA-specific cells to express antiinflammatory cytokines and neurotrophic factor in the organ in which the pathological processes occur correlates directly with the therapeutic activity of GA in experimental autoimmune encephalomyelitis/multiple sclerosis.

Topics: Adoptive Transfer; Animals; Brain; Brain-Derived Neurotrophic Factor; Cytokines; Encephalomyelitis, Autoimmune, Experimental; Female; Glatiramer Acetate; Humans; Immunohistochemistry; Immunosuppressive Agents; Interferon-gamma; Interleukin-10; Mice; Mice, Inbred BALB C; Multiple Sclerosis; Peptides; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer; Th2 Cells; Transforming Growth Factor beta

During mammalian central nervous system (CNS) development, contact-mediated activation of Notch1 receptors on oligodendrocyte precursors by the ligand Jagged1 induces Hes5, which inhibits maturation of these cells. Here we tested whether the Notch pathway is re-expressed in the adult CNS in multiple sclerosis (MS), an inflammatory demyelinating disease in which remyelination is typically limited. We found that transforming growth factor-beta 1 (TGF-beta 1), a cytokine upregulated in MS, specifically re-induced Jagged1 in primary cultures of human astrocytes. Within and around active MS plaques lacking remyelination, Jagged1 was expressed at high levels by hypertrophic astrocytes, whereas Notch1 and Hes5 localized to cells with an immature oligodendrocyte phenotype, and TGF-beta 1 was associated with perivascular extracellular matrix in the same areas. In contrast, there was negligible Jagged1 expression in remyelinated lesions. Experiments in vitro showed that Jagged1 signaling inhibited process outgrowth from primary human oligodendrocytes. These data are the first to implicate the Notch pathway in the limited remyelination in MS. Thus, Notch may represent a potential target for therapeutic intervention in this disease.

Topics: Animals; Astrocytes; Calcium-Binding Proteins; Cells, Cultured; Humans; Intercellular Signaling Peptides and Proteins; Jagged-1 Protein; Membrane Proteins; Multiple Sclerosis; Myelin Sheath; Oligodendroglia; Oligonucleotide Array Sequence Analysis; Proteins; Receptor, Notch1; Receptors, Cell Surface; Serrate-Jagged Proteins; Signal Transduction; Transcription Factors; Transforming Growth Factor beta; Transforming Growth Factor beta1

Genome screenings in multiple sclerosis (MS) have identified multiple susceptibility regions supporting a polygenic model for this disease. Evidence for linkage was consistently observed at ch.19q13 suggesting the presence of an MS gene(s) in this region. Several interesting candidate genes are encoded within this region, including transforming growth factor-beta 1 (TGFB1) and interleukin-11 (IL11). Both are multifunctional cytokines with significant and well-characterized immunomodulatory properties. We performed a comprehensive evaluation of common polymorphisms within the TGFB1 and IL11 loci and three closely flanking microsatellite markers (D19S421, CEA, D19S908) in 161 stringently ascertained and clinically characterized MS multiplex families using tests of both linkage (lod score, sib-pair analysis) and association (pedigree disequilibrium test or PDT). Patients and families were stratified by HLA-DR2 status to search for two-locus interactions. Suggestive evidence for linkage and association to CEA (lod score = 1.25, theta = 0.20, p = 0.015, respectively), located 0.4 cM from TGFB1, was observed in DR2 positive families only. Distinct clinical phenotypes were also examined and an association between a TGFB1 haplotype and a mild disease course was present (p = 0.008), raising the possibility that TGFB1 or a nearby locus may influence disease expression.

Topics: Adult; Amino Acid Sequence; Base Sequence; Chromosome Mapping; Disability Evaluation; Female; Genetic Linkage; Genetic Predisposition to Disease; Genotype; Haplotypes; Humans; Interleukin-11; Male; Microsatellite Repeats; Middle Aged; Multiple Sclerosis; Polymorphism, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1

Little is known about the involvement of cytokines in the pathogenesis of primary progressive (PP) multiple sclerosis (MS). We evaluated in this cross-sectional study whether IL-18, IL-12p35, IL-12p40, TNF-alpha, IFN-gamma, IL-10, IL-4, TGF-beta, IL-12Rbeta1, and IL-12Rbeta2 mRNA expression in unstimulated white blood cells showed significant differences between relapsing-remitting (RR), secondary progressive (SP) and PP MS patients, and healthy controls. All clinical subtypes showed unique mRNA expression patterns as compared to the controls. Both RR and SP patients displayed increased levels of IL-12p40, IL-18, and TGF-beta mRNA compared to controls, whereas PP patients showed only increased IL-18 mRNA levels. Both in PP and SP patients, IFN-gamma and IL-10 mRNA were decreased compared to RR patients and controls. PP patients were unique in that they showed decreased IL-12Rbeta1 mRNA. In conclusion, our data show that the assessment of cytokine (receptor) mRNA profiles is useful to discriminate between the different clinical subtypes and suggest that different cytokines are involved in the pathogenesis of PP MS as compared to RR and SP MS.

Topics: Adult; Cross-Sectional Studies; Cytokines; Disease Progression; Female; Humans; Interleukin-10; Interleukin-12; Interleukin-18; Leukocytes; Male; Middle Aged; Multiple Sclerosis; Receptors, Interleukin; Receptors, Interleukin-12; RNA, Messenger; Transforming Growth Factor beta

Transforming growth factor beta1 (TGFbeta1) is a Th2 cytokine encoded on chromosome 19q13, a region possibly linked to multiple sclerosis (MS). TGFbeta1 exerts favorable effects on experimental allergic encephalomyelitis. We performed a comprehensive search for genetic variants in this gene in 122 population-based sporadic cases of MS. We detected six variants, including three missense variants. We tested for association of the variants with susceptibility and course of MS and for linkage and transmission disequilibrium in a family series consisting of 395 samples in 59 pedigrees. Genetic variation in TGFB1 does not appear to contribute in a major way to susceptibility to MS.

Topics: Adult; Chromosomes, Human, Pair 19; Exons; Female; Gene Frequency; Genetic Predisposition to Disease; Genetic Testing; Genetic Variation; Genotype; Haplotypes; Humans; Linkage Disequilibrium; Male; Middle Aged; Multiple Sclerosis; Mutation, Missense; Polymorphism, Genetic; Promoter Regions, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1

TGF-beta 2 is a potent immunoregulatory mediator that influences B cell, T cell, and macrophage function. To test whether this cytokine alters pathology in a model of virus-induced demyelinating disease, we treated SJL/J mice with TGF-beta 2 either before or after infection with Theiler's murine encephalomyelitis virus. Treatment continued three times weekly through day 35 postinfection. TGF-beta 2 administration resulted in significantly smaller lesions and fewer virus Ag-positive cells in the spinal cords of infected SJL/J mice. Mice treated with TGF-beta 2 had similar levels of virus-specific IgG as infected, control-treated mice. TGF-beta 2 administration significantly increased the level of non-virus-specific activated CTLs, but had no effect on virus-specific CTLs. TUNEL revealed a decrease in the number of apoptotic nuclei in the spinal cord white matter of mice treated in vivo with TGF-beta 2. Immunostaining with an Ab to F4/80 revealed that TGF-beta 2-treated mice had significantly fewer F4/80-positive cells in the white matter of the spinal cord as compared with infected control-treated mice. These data suggest that TGF-beta 2 may control virus-induced demyelination via an immunomodulatory mechanism that reduces macrophage infiltration.

Topics: Animals; Antigens, Viral; Apoptosis; Cardiovirus Infections; Cell Count; Cytotoxicity Tests, Immunologic; Disease Models, Animal; Disease Susceptibility; Female; Injections, Intraperitoneal; Macrophages; Mice; Mice, Inbred Strains; Multiple Sclerosis; Myelin Sheath; Spinal Cord; T-Lymphocytes, Cytotoxic; Theilovirus; Transforming Growth Factor beta; Viral Plaque Assay; Virus Replication

In cerebral malaria (CM), pathologic cytokine expression patterns are thought to contribute to disruption of the blood-brain barrier, inflammation, and astrocytic scar formation. Expression of transforming growth factor (TGF)-beta1, -beta2, and -beta3 was analyzed in the brains of 7 patients who died with CM and in 8 control patients. In the brains of patients with CM, there were significantly (P=.0003) more TGF-beta1-immunoreactive astrocytes adjacent to brain vessels with deposition of malarial pigment, significantly (P=.0081) more TGF-beta2-expressing macrophages/microglial cells in glioses of ring hemorrhages and Dürck's granulomas, and significantly (P=.0022) more TGF-beta3-expressing smooth-muscle cells and endothelial cells of brain vessels with sequestration. It is concluded that focal accumulation of TGF-beta1, -beta2, and -beta3 provides evidence for their involvement in the reorganization process of the brain parenchyma, immunologic dysfunction, and endothelial cell activation in patients with CM.

Topics: Alzheimer Disease; Brain; Humans; Malaria, Cerebral; Meningitis, Bacterial; Multiple Sclerosis; Protein Isoforms; Transforming Growth Factor beta

The immunological effects of high-dose methylprednisolone in attacks of multiple sclerosis and acute optic neuritis have only been examined in a few randomized, controlled trials. We studied immunological changes in 50 patients with optic neuritis or multiple sclerosis who underwent lumbar puncture before and 1 week after completing a 15-day course of oral high-dose methylprednisolone treatment. Treatment resulted in a decrease in the concentration of myelin basic protein, a decrease in the serum concentration of immunoglobulin G (IgG) and intrathecal IgG synthesis, an increase in the cerebrospinal fluid concentration of transforming growth factor-beta1, and changes in the expression of CD25, CD26, and human leukocyte antigen-DR (HLA-DR) on CD4 T-cells. No effect was seen on the cerebrospinal fluid leucocyte count or the cerebrospinal fluid activity of matrix metalloproteinase-9 (MMP-9). The lack of a persistent effect on cerebrospinal fluid leucocyte recruitment and MMP-9 activity, despite changes in IgG synthesis, T-cell activation, and cytokine production, suggests that modulation of the function of inflammatory cells may contribute to the clinical efficacy of oral high-dose methylprednisolone treatment in optic neuritis and multiple sclerosis.

Topics: Acute Disease; Administration, Oral; Adult; Anti-Inflammatory Agents; Antigens; CD4-Positive T-Lymphocytes; Dose-Response Relationship, Drug; Female; Flow Cytometry; Humans; Immune System; Immunoglobulin G; Leukocyte Count; Male; Matrix Metalloproteinase 9; Methylprednisolone; Multiple Sclerosis; Myelin Basic Protein; Optic Neuritis; Randomized Controlled Trials as Topic; Spinal Cord; Spinal Puncture; Transforming Growth Factor beta

Topics: Animals; Bone Diseases; Extracellular Matrix Proteins; Gene Expression Regulation; Genes, Tumor Suppressor; Humans; Inflammation; MAP Kinase Signaling System; Multiple Sclerosis; Signal Transduction; Transforming Growth Factor beta

It is known that the pleiotropic cytokine transforming growth factor beta (TGF-beta) has a regulatory role in the process of tissue repair and remodelling following injury. As reports on these molecules in multiple sclerosis (MS) lesion with different lesional activity are rare, we studied the cellular localization of TGF-beta1, -beta2, and -beta3 isoforms, and TGF-beta receptor type I (TGF-betaR-I) and TGF-betaR-II expression by immunohistochemistry on postmortem brain tissue from MS and normal control cases. To validate the TGF-beta staining results we demonstrated that cultured human adult astrocytes that produce biological active TGF-beta2, and to a lesser extent TGF-beta1, were immunoreactive for all 3 TGF-beta isoforms. Moreover, at mRNA level TGF-beta1 was detected in MS and normal control brain tissue. In normal control brain tissue, TGF-beta isoforms were expressed in ramified microglia and TGF-beta2, and -beta3 on neuronal cells in the gray matter TGF-betaR-I and TGF-betaR-II expression was found on endothelial cells, astrocytes, microglia, and neurons. In active demyelinating MS lesions a strong to intense immunoreactivity was detected for all 3 TGF-beta isoforms in perivascular and parenchymal (foamy) macrophages and in hypertrophic astrocytes. Strong immunoreactivity for TGF-betaR-I and TGF-betaR-II was found on macrophages in both parenchymal and perivascular areas and on hypertrophic astrocytes and endothelial cells in active demyelinating MS lesions. In chronic active and inactive MS lesions, all 3 TGF-beta isoforms and their receptors were strongly expressed in hypertrophic astrocytes. Our findings strongly suggest that the expression of the various TGF-beta isoforms and their receptor types found in MS lesions with different cellular activity participate in reactive processes leading to the formation of chronic MS lesions.

Topics: Activin Receptors, Type I; Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Astrocytes; Brain Chemistry; Cells, Cultured; Cerebellum; Corpus Callosum; DNA Primers; Female; Gene Expression; Humans; Isomerism; Macrophages; Male; Microglia; Middle Aged; Multiple Sclerosis; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta

Topics: Animals; Apoptosis; Arthritis, Experimental; Arthritis, Rheumatoid; Autoimmune Diseases; Celiac Disease; Diabetes Mellitus, Type 1; Genetic Predisposition to Disease; Humans; Interleukin-10; Mice; Mice, Inbred MRL lpr; Mice, Knockout; Multiple Sclerosis; Risk Factors; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

We amplified the mRNA for cytokines in peripheral blood mononuclear cells (PBMC) and cerebrospinal fluid (CSF) cells from 18 multiple sclerosis (MS) patients and 21 other neurological patients, using the reverse transcription polymerase chain reaction (RT-PCR). Radioactive hybridization of the amplified DNA allowed quantitation of mRNA levels. Expression of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma and interleukin (IL)-10 mRNA was elevated in CSF cells from MS patients. IFN-gamma and IL-10 mRNA levels were higher in MS patients than in other inflammatory neurological diseases. mRNA coding for transforming growth factor (TGF)-beta was detectable in the majority of cases, with higher expression in CSF cells of MS and other inflammatory neurological diseases than in noninflammatory controls, and higher expression in PBMC of MS patients than in all other cases. In many MS patients both proinflammatory and immunoregulatory cytokine messages were detected in the CSF compartment without correlation with the clinical activity of the disease. In contrast, mRNA for the costimulatory molecule B7.1 was only detected in the CSF cells of some MS patients, who showed clinical signs of acute relapse at the time of the spinal tap.

Topics: Cytokines; Humans; Interferon-gamma; Interleukins; Leukocytes, Mononuclear; Multiple Sclerosis; Reverse Transcriptase Polymerase Chain Reaction; Stimulation, Chemical; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Eight relapsing-remitting multiple sclerosis (MS) patients were tested for the level of transforming growth factor beta1 (TGFbeta1) mRNA in peripheral blood mononuclear cells every 15 days for 6 months. Disease activity was evaluated every 4 weeks by magnetic resonance imaging (MRI) and neurological examination. An inverse correlation was found between the level of TGFbeta1 mRNA and MRI disease activity. The level of TGFbeta1 mRNA predicted the presence of disease activity in the scans performed 2-4 weeks later with high sensitivity (88%) and specificity (87.5%) suggesting that TGFbeta1 mRNA quantification could be an indicator of disease activity in MS.

Topics: Adult; Brain; Female; Gadolinium DTPA; Humans; Leukocytes, Mononuclear; Magnetic Resonance Imaging; Male; Multiple Sclerosis; Recurrence; RNA, Messenger; Statistics, Nonparametric; Transcription, Genetic; Transforming Growth Factor beta

To examine the influence of TGF-beta genes on MS susceptibility.. TGF-beta, of which three homologous isoforms exist (1, 2 and 3), is a strongly immunosuppressive cytokine-inhibiting expression of pro-inflammatory cytokines and blocking cytokine induction of adhesion molecules. TGF-beta delays onset of EAE and TGF-beta 1 gene knockout mice develop fatal multifocal inflammatory disease. High TGF-beta levels exist during MS remission whilst E-selectin, whose expression is inhibited by TGF-beta, is found at higher levels in primary progressive disease (PPMS) and it is postulated that the unremitting course of PPMS may be due to low levels of TGF-beta.. Gene association studies using separate polymorphic microsatellite markers for TGF-beta 1 and TGF-beta 2 were performed, incorporating 151 relapsing-remitting or secondary progressive MS (RR/SPMS) patients, 104 PPMS patients and 159 normal controls (Nor). Forward primers were 5' end-labelled with 6-Fam, PCR products were analysed on an Applied Biosystems 373A fluorescent fragment analyser and Genescan 672 software was used for allele sizing.. No significant differences existed in allele frequencies between either MS group and controls regarding the TGF-beta 1 marker: RR/SPMS vs Nor (P = 0.48, df = 8); PPMS vs Nor (P = 0.34, df = 8). Similarly there were no associations demonstrated with the TGF-beta 2 marker: RR/SPMS vs Nor (P = 0.24, df = 2); PPMS vs Nor (P = 0.53, df = 2).. These data indicate that TGF-beta 1 and beta 2 genes are not loci influencing MS susceptibility, either RR/SPMS or PPMS, in this population.

Topics: Alleles; DNA, Satellite; Female; Genetic Linkage; Genetic Markers; Heterozygote; Humans; Male; Multiple Sclerosis; Northern Ireland; Polymorphism, Genetic; Transforming Growth Factor beta

Cytokines have a crucial role in initiation and perturbation of EAE that represents an animal model of multiple sclerosis (MS). Administration of transforming growth factor-beta1 (TGF-beta1) to EAE mice improves clinical EAE and prevents relapses by unknown mechanisms. Administering low doses of TGF-beta1 nasally, we confirmed that TGF-beta1 inhibited development and relapse of protracted-relapsing EAE (PR-EAE) in DA rats. Infiltration of CD4+ T-cells and macrophages within the central nervous system was clearly reduced, while proliferation and IFN-gamma secretion of mononuclear cells (MNC) was augmented in TGF-beta1-treated EAE rats compared to PBS-treated control EAE rats. TGF-beta1 administered nasally also increased nitric oxide production and CD4+ T cell apoptosis. TGF-beta1 treated rats showed augmented proliferation of dendritic cells (DC) compared to MNC. These data imply that low doses of TGF-beta1 given by the nasal route prevent PR-EAE and upregulate DC functions that may be involved for disease prevention.

Topics: Administration, Intranasal; Animals; Apoptosis; CD4 Lymphocyte Count; Cell Division; Cells, Cultured; Data Interpretation, Statistical; Dendritic Cells; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunohistochemistry; Immunotherapy; Male; Mice; Multiple Sclerosis; Myelin Basic Protein; Nitric Oxide; Rats; Spleen; Transforming Growth Factor beta; Up-Regulation

Canine distemper virus (CDV) infection in dogs is commonly associated with demyelinating leukoencephalitis (DL). Although the mechanism of primary demyelination in distemper remains undetermined recent studies showed a direct virus-induced cytolysis in early non-inflammatory and immune-mediated mechanisms in inflammatory lesions. To further investigate the pathogenesis of this morbillivirus-induced demyelination the expression of a variety of cytokine mRNA species (interleukin (IL)-1beta, IL-2, IL-6, IL-10, IL-12, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, and interferon (IFN)-gamma in cerebrospinal fluid cells of 12 dogs with CDV encephalitis was investigated employing reverse transcription-polymerase chain reaction (RT-PCR) and these findings were correlated to the type of CNS lesions. Neuropathology revealed the whole spectrum of distemper DL lesions from acute to chronic alterations, however, most plaques lacked active demyelination. Three control animals were devoid of any cytokine expression, whereas in distemper animals IL-10 transcripts were found in nine dogs with acute and chronic lesions. IL-6, TNF, and TGF mRNA was found in six, four, and three animals, respectively. IL-12 and IFN-gamma, suggestive of a TH1-like dominated immune response, were detected only in one animal with chronic lesions. Summarized, TNF and IL-6, associated with disease exacerbation, and IL-10 and TGF, indicative of remission, were often observed simultaneously in distemper DL and could not be assigned to a specific disease stage. However IL-10 mRNA remained the most frequently detected cytokine indicating a stage of inactivity in most animals investigated.

Topics: Animals; Brain; Demyelinating Diseases; Disease Models, Animal; Distemper; Distemper Virus, Canine; DNA Primers; Dogs; Encephalitis, Viral; Gene Expression; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-2; Interleukin-6; Multiple Sclerosis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The mechanisms by which IFNbeta-1b acts in the treatment of multiple sclerosis (MS) patients are not completely known. We investigated the influence of IFNbeta-1b on the mRNA-expression of the immunosuppressive cytokine TGFbeta-1 and the proinflammatory mediator TNFalpha in an in vitro model by the method of non-radioactive in situ hybridization. Peripheral blood lymphocytes (PBL) were isolated from eight patients with relapsing remitting form of MS during remission and from six healthy controls. They were stimulated with IFNbeta-1b in different concentrations for 24 h. In both groups a statistically significant dose-dependent increase of TGFbeta-1-mRNA and decrease of TNFalpha-mRNA was demonstrable in the cultured stimulated blood lymphocytes compared to unstimulated cells. Stimulations with lipopolysaccharide (LPS) led to an increase of both cytokine-mRNAs in the lymphocytes. These data suggest specific and dose-dependent effects of IFNbeta-1b and hint at immunomodulatory properties of this drug to regulate the cytokine dysbalance in MS. This might be one mechanism by which IFNbeta-1b mediates its beneficial effects on the course of the disease.

Topics: Adjuvants, Immunologic; Adult; Dose-Response Relationship, Drug; Female; Gene Expression; Humans; In Vitro Techniques; Interferon-beta; Lipopolysaccharides; Lymphocytes; Male; Middle Aged; Multiple Sclerosis; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system and one of the earliest changes in inflammatory focus involves the activation of vascular endothelial cells. We determined the plasma level of plasminogen activator inhibitor-1 (PAI-1), a key regulator of fibrinolysis and cell migration, in patients with MS. The level of plasma PAI-1 was significantly higher in active MS cases when compared to stable MS and controls. Plasma concentrations of tissue plasminogen activator, transforming growth factor beta-1, and lipoprotein-a remained normal in spite of disease activity. These results suggested that PAI-1 plasma levels are associated with MS disease activity and is a good marker for MS relapse.

Topics: Adult; Biomarkers; Disease Progression; Female; Humans; Immunoglobulin G; Lipoprotein(a); Male; Middle Aged; Multiple Sclerosis; Plasminogen Activator Inhibitor 1; Recurrence; Reference Values; Tissue Plasminogen Activator; Transforming Growth Factor beta

The mechanisms by which corticosteroids act in the treatment of an acute relapse in multiple sclerosis (MS) are not completely known. We investigated the mRNA and protein expression of transforming-growth-factor-beta1 (TGFbeta1), a cytokine with anti-inflammatory and immunosuppressive potentials, in peripheral blood mononuclear cells (PBMC) and serum of 10 patients with an acute relapse of MS before, during and after the treatment with 500 mg prednisolone daily over 5 days. The expression of TGFbeta1-mRNA increased at day 3-5 and declined at day 8-10. Serum levels of TGFbeta1 demonstrated a comparable course. The present data suggest that corticosteroids induce the expression of TGFbeta1 in vivo. This is might be an other mechanism by which corticosteroids mediate immunosuppression.

Topics: Adult; Anti-Inflammatory Agents; Female; Humans; Leukocytes, Mononuclear; Male; Multiple Sclerosis; Prednisolone; RNA, Messenger; Transforming Growth Factor beta

Several genetic factors are likely to play a role in the etiology of multiple sclerosis (MS). We used a candidate gene strategy in a study of polymorphic markers within or close to genes encoding cytokines (interferon-gamma (IFN-gamma), interleukin-2 (IL-2), IL-4, IL-4 receptor (IL-4R), IL-10, transforming growth factor-beta1 and -beta2) and myelin proteins (2',3'-cyclic-nucleotide 3'-phosphohydrolase (CNP:ase), myelin associated glycoprotein, oligodendrocyte myelin glycoprotein, proteolipid protein) in 34 Swedish multiplex MS families and in 147 sporadic MS patients and 95 healthy controls. No evidence for linkage was observed in two-point linkage analysis. However, a slightly positive LOD score of 0.88 (theta = 0.01) for IFN-gamma was found. Affected pedigree member (APM) analysis indicated a possible linkage with TGF-beta2 (p = 0.008) and IL-4R (p = 0.043). None of the cytokine markers were associated with MS in case-control analysis. Our results suggest a possible importance of the TGF-beta2, IL-4R and IFN-gamma genes in MS.

Topics: 2',3'-Cyclic-Nucleotide Phosphodiesterases; Case-Control Studies; Cytokines; Family Health; Genetic Linkage; GPI-Linked Proteins; Humans; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-4; Multiple Sclerosis; Myelin Proteins; Myelin Proteolipid Protein; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Polymorphism, Genetic; Receptors, Interleukin-4; Transforming Growth Factor beta

Multiple sclerosis is postulated to be a Th1-type cell-mediated autoimmune disease. We investigated cytokine profiles in patients with progressive multiple sclerosis by using intracytoplasmic staining. We found increased IL-12 production by monocytes and increased IFN-gamma production by T cells in untreated patients as compared with controls. In patients treated with methotrexate, methylprednisolone, or cyclophosphamide/methylprednisolone (CY/MP), only CY/MP treatment normalized the elevated IL-12 production. Furthermore, CY/MP-treated patients had decreased IFN-gamma and increased IL-4, IL-5, and TGF-beta expression. Patients followed prospectively before and after starting CY/MP treatment showed a gradual decrease in IL-12 and IFN-gamma production and an increase in IL-4 and IL-5. In vitro, addition of 4-hydroperoxycyclophosphamide, a metabolite of cyclophosphamide decreased IL-12 production in mononuclear cell cultures. When patients were classified as having active or stable disease, IL-12 production correlated with disease activity. In summary, our results demonstrate a Th1-type cytokine bias in peripheral blood mononuclear cells of untreated progressive MS patients that is reversed by CY/MP treatment and is associated with Th2 and TGF-beta (Th3) type responses. These findings provide a basis for immune monitoring of patients with MS and suggest that treatments that downregulate IL-12 may prove to be beneficial in progressive MS.

Topics: Cyclophosphamide; Female; Humans; Interferon-gamma; Interleukin-2; Leukocytes, Mononuclear; Male; Middle Aged; Monocytes; Multiple Sclerosis; Prospective Studies; T-Lymphocytes; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

Levels of tumor necrosis factor (TNF)-alpha, granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-10 and transforming growth factor (TGF)-beta in cerebrospinal fluid (CSF) and serum of 29 patients with multiple sclerosis (MS) of the relapsing-remitting type and of 20 controls with other non inflammatory neurological diseases were studied. Sixteen patients were in the active phase of disease and 13 in remission. In CSF, higher IL-10 and TGF-beta concentrations were found in patients with a stable phase of MS, while in the active phase there were elevated levels of TNF-alpha and GM-CSF. These results suggest that a different cytokine pattern could be probably involved in the pathogenesis of relapsing-remitting MS.

Topics: Adolescent; Adult; Cytokines; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-10; Male; Middle Aged; Multiple Sclerosis; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Steroid hormones have long been known to modulate immune function, and recent studies indicate that one of the means by which they do so involves effects on the secretion of immunoregulatory cytokines. Our laboratory has found recently that estradiol (E2) selectively modifies cytokine secretion in proteolipid protein (PLP)-specific, CD4+ T cell clones isolated from patients with the demyelinating disease, multiple sclerosis, and from normal control subjects. The data suggest that E2 may play a role in regulating the balance between pro- and antiinflammatory conditions, especially at concentrations typical of pregnancy. To determine whether other pregnancy-associated steroid hormones are capable of similar activity, we expanded our testing to include estrone (E1), estriol (E3), progesterone, and dexamethasone. The results indicate that E1 and E3 enhance secretion of Ag- or anti-CD3-stimulated IL-10 and IFN-gamma in dose-dependent fashion, almost identical to that of E2. The effect on IL-10 was more potent than occurred with IFN-gamma. In addition, E1 and E3, like E2, had a biphasic effect on TNF-alphabeta secretion, with low concentrations stimulatory, and high doses inhibitory. None of the estrogens influenced IL-4 or TGF-beta secretion. Progesterone enhanced secretion of IL-4, without affecting any other tested cytokine. Finally, dexamethasone induced TGF-beta secretion, but inhibited IFN-gamma and TNF-alphabeta. This differential effect of steroid hormones on the secretion of cytokines by CD4+ human T cell clones is consistent with the possibility that, collectively, they promote antiinflammatory conditions at high concentrations typical of pregnancy.

Topics: CD4-Positive T-Lymphocytes; Cell Separation; Clone Cells; Cytokines; Dexamethasone; Estriol; Estrone; Female; Hormones; Humans; Male; Multiple Sclerosis; Myelin Proteolipid Protein; Peptide Fragments; Progesterone; Transforming Growth Factor beta

The mechanisms by which interferon-beta-1b (IFNbeta-1b) acts in the treatment of patients with multiple sclerosis (MS) are not completely known. A total of 10 MS patients were treated with 8 million units of IFNbeta-1b every other day. Compared to baseline and control group the expression of TGFbeta-1-mRNA by PBMC was persistently increased at week 6, month 3 and month 6 (p < or = 0.04), that of the TGFbeta-1 receptor type II from day 5 up to month 6 (p < 0.01). The mRNA and protein expression of tumor necrosis factor-alpha (TNFalpha)-receptor (55 kDa) was only temporarily elevated at the beginning of the therapy. Serum levels of sVCAM were increased during the whole time of treatment (p < 0.01). The CD8CD38 lymphocyte subpopulation was continuously elevated from day 5 up to month 6 (p < 0.01). No persistently significant changes were demonstrable concerning the percentage of total CD4, CD8, CD19 or in CD4 subpopulations (CD4CD29, CD4CD45RA). The present data suggest that IFNbeta-1b induces the expression of TGFbeta-1- and TGFbeta-R-II-mRNA by PBMC and increases levels of sVCAM-1 and of circulating activated CD8 cells (CD8CD38) in serum. These might be other mechanisms by which IFNbeta-1b mediates its positive effects in the treatment of MS patients.

Topics: Adult; Antigens, CD19; B-Lymphocytes; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; DNA, Complementary; Female; Flow Cytometry; Gene Expression Regulation; Humans; Interferon-beta; Leukocyte Common Antigens; Lymphocyte Subsets; Male; Multiple Sclerosis; Neuroimmunomodulation; Receptors, Transforming Growth Factor beta; Receptors, Tumor Necrosis Factor; RNA, Messenger; Transforming Growth Factor beta; Vascular Cell Adhesion Molecule-1

Costimulatory molecules B7-1 (CD80) and B7-2 (CD86) are differently involved in T cell stimulation. In chronic experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS), B7-1 was preferentially involved in pathophysiology of relapses. We used reverse transcription polymerase chain reaction (RT-PCR) to amplify the mRNA coding for these molecules in cerebrospinal fluid (CSF) cells and peripheral blood mononuclear cells (PBMC) from 18 MS patients and 21 other neurological patients. In CSF cells of MS cases, B7-1 mRNA was only detected in some patients who showed clinical signs of acute relapse at the time of the spinal tap, while B7-2 mRNA was widely detectable without difference between active or stable MS and controls. mRNA coding for transforming growth factor-beta (TGF-beta) was detectable in the majority of cases, with higher expression in CSF cells of MS and other inflammatory neurological diseases (OIND) than in noninflammatory controls, and higher expression in PBMC of MS patients than in all other cases. Finally, mRNA coding for interleukin (IL)-12p40 was only detected in a very few number of MS and inflammatory cases. These findings were related to previous detection of other cytokines in the same cases, showing relationships in CSF cells between high expression of B7-1, IL-12p40 and TNF-alpha.

Topics: Actins; Antigens, CD; B7-1 Antigen; B7-2 Antigen; Cerebrospinal Fluid; Gene Expression; Humans; Interleukin-12; Leukocytes, Mononuclear; Membrane Glycoproteins; Multiple Sclerosis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta

The level of bioactive transforming growth factor-beta (TGF-beta) was measured in serum from patients with chronic fatigue syndrome (CFS), healthy control subjects, and patients with major depression, systemic lupus erythematosis (SLE), and multiple sclerosis (MS) of both the relapsing/remitting (R/R) and the chronic progressive (CP) types. Patients with CFS had significantly higher levels of bioactive TGF-beta levels compared to the healthy control major depression, SLE, R/R MS, and CP MS groups (P < 0.01). Additionally, no significant differences were found between the healthy control subjects and any of the disease comparison groups. The current finding that TGF-beta is significantly elevated among patients with CFS supports the findings of two previous studies examining smaller numbers of CFS patients. In conclusion, TGF-beta levels were significantly higher in CFS patients compared to patients with various diseases known to be associated with immunologic abnormalities and/or pathologic fatigue. These findings raise interesting questions about the possible role of TGF-beta in the pathogenesis of CFS.

Topics: Depression; Factor Analysis, Statistical; Fatigue Syndrome, Chronic; Humans; Lupus Erythematosus, Systemic; Multiple Sclerosis; Transforming Growth Factor beta

Topics: Administration, Oral; Animals; Cell Line; Cytokines; Humans; Multiple Sclerosis; Myelin Basic Protein; T-Lymphocytes; Transforming Growth Factor beta

Cerebral expression of the injury response cytokine transforming growth factor-beta 1 (TGF-beta 1) has been found to be increased in several neurological diseases but it remains unclear whether its function is primarily beneficial or detrimental. Here we show that transgenic (tg) mice that overexpress bioactive (TGF-beta 1 in the central nervous system (CNS) and show no overt phenotype in the unmanipulated state, are more susceptible to the immune-mediated CNS disease experimental autoimmune encephalomyelitis (EAE). TGF-beta 1 tg mice with EAE showed an earlier onset of clinical symptoms, more severe disease and increased mononuclear cell infiltration in their spinal cords compared with non-tg littermate controls with EAE. Whereas previous observations indicated that increased peripheral levels of TGF-beta 1 can suppress EAE, our findings demonstrate that local expression of TGF-beta 1 within the CNS parenchyma can enhance immune cell infiltration and intensify the CNS impairment resulting from peripherally triggered autoimmune responses.

Topics: Animals; Antigens; Astrocytes; Brain; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Glial Fibrillary Acidic Protein; Macrophages; Male; Mice; Mice, Inbred BALB C; Mice, Transgenic; Multiple Sclerosis; Mutagenesis; Spinal Cord; Transforming Growth Factor beta

The polypeptide transforming growth factor-beta 1 (TGF-beta 1) is a product of activated monocytes, among other inflammatory cells, and it affects immune responsiveness, cellular growth and differentiation. TGF-beta 1 has potent T-cell inhibiting activities. It may play an important role in limiting autoimmune inflammation. We were interested about levels of biologically active and total TGF-beta 1 in serum and CSF in patients suffering from multiple sclerosis.. We measured biologically active and total TGF-beta 1 in serum and CSF using ELISA-technique in 64 MS patients with 57 during acute exacerbation of MS and 7 in remission (primary-relapsing: n = 59; primary-progressive: n = 5), 20 healthy subjects, and 21 patients with other non-inflammatory neurological diseases (OND).. Biologically active TGF-beta 1 in serum was reduced in MS patients compared to controls, on the other hand total TGF-beta 1 was elevated in CSF compared to patients with OND. Biologically active TGF-beta 1 in CSF correlated positively with the duration of the acute relapse in patients with primary-relapsing MS. The more relapses the patients had the higher was biologically active TGF-beta 1 in CSF. Total TGF-beta 1 in CSF correlated with macrophages in CSF and albumin quotient.. We found that an elevated level of biologically active TGF-beta 1 in CSF might be useful as an indicator of disease limitation while active TGF-beta 1 in serum is reduced in multiple sclerosis. Measuring TGF-beta 1 in body fluids by ELISA techniques produces valid results and might be used for further studies focusing on the role of this cytokine in MS.

Topics: Adult; Age of Onset; Analysis of Variance; Biomarkers; Case-Control Studies; Chi-Square Distribution; Disease Progression; Enzyme-Linked Immunosorbent Assay; Female; Humans; Male; Multiple Sclerosis; Severity of Illness Index; Time Factors; Transforming Growth Factor beta

Cerebrospinal fluid (CSF) and serum levels of transforming growth factor (TGF)-beta and soluble intercellular adhesion molecule-1 (sICAM-1) were evaluated in ten patients with definite multiple sclerosis (MS) of the relapsing-remitting type. CSF TGF-beta levels of MS patients in remission were significantly (p < 0.01) higher than of MS patients in active phase, and there was a significant inverse correlation (p < 0.05) between TGF-beta and slCAM-1 levels in the CSF of patients in both remitting and relapsing type. This is consistent with a possible down-regulation of TGF-beta on ICAM-1 expression and suggests a possible synthesis in the central nervous system of TGF-beta.

Topics: Adult; Enzyme-Linked Immunosorbent Assay; Female; Humans; Intercellular Adhesion Molecule-1; Male; Middle Aged; Multiple Sclerosis; Recurrence; Transforming Growth Factor beta

Transforming growth factor (TGF)beta plays a role in injury repair in sites surrounding brain injury. The present study tested the hypothesis that TGFbeta1 and TGFbeta2 levels in the postmortem CSF of patients with neurodegenerative disorders would be elevated compared to those in normal subjects. Free TGFbeta1 and total TGFbeta2 were measured by ELISA in postmortem ventricular cerebrospinal fluid (vCSF) of patients with Parkinson's disease (n = 30), Alzheimer's disease (n = 30), multiple sclerosis (n = 15), and schizophrenia (n = 12) and of normal controls (n = 16). In addition, albumin, IgG, and total protein in vCSF were measured. Both TGFbeta1 and TGFbeta2 were significantly different between groups (P < 0.002 and P < 0.001, respectively). Parkinson's disease vCSF showed significant increases in both TGFbeta1 (P = 0.015) and TGFbeta2 (P = 0.012) compared to normal controls. There was a trend for TGFbeta2 to be elevated in Alzheimer's disease and multiple sclerosis vCSFs, which failed to achieve significance. There were no differences between controls and schizophrenics in TGFbeta1 or TGFbeta2. Alzheimer's disease vCSF showed a significant decrease in protein compared to all other groups, which was not related to blood-brain barrier permeability, age, or autolysis differences. Evidence is presented suggesting that some TGFbeta1 may leak into the vCSF from plasma. Autopsy vCSF levels of TGFbeta isoforms were found to be distinctly different from those reported for human serum, especially for TGFbeta2, which is undetectable in plasma. These results indicate that further in vivo studies of TGFbeta2 in the CSF of Parkinson's disease patients are warranted to determine the relationship between clinical status, medication, and TGFbeta2 concentrations.

Topics: Age Factors; Aged; Aged, 80 and over; Albumins; Alzheimer Disease; Blood-Brain Barrier; Cerebral Ventricles; Cerebrospinal Fluid Proteins; Cross Reactions; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin G; Male; Middle Aged; Multiple Sclerosis; Parkinson Disease; Schizophrenia; Sex Factors; Transforming Growth Factor beta

We have recently isolated a panel of T-cell clones from chronic progressive multiple sclerosis (MS) patients that are capable of functioning as antigen-presenting cells and of expressing the costimulatory molecules B7-1 and B7-2. In this report we show that these T-cell clones are resistant to inhibitory regulation, including the induction of anergy and sensitivity to tumor growth factor-beta (TGF-beta)-induced growth inhibition. The resistance to anergy induction was associated with expression of B7 costimulatory molecules. These data suggest that lack of responsiveness to peripheral inhibitory signals may account for the entry of autoimmune diseases into a chronic progressive phase.

Topics: Adult; Antibodies, Monoclonal; Antigen-Presenting Cells; B7-1 Antigen; Cell Division; Chronic Disease; Clonal Anergy; Female; Histocompatibility Testing; Humans; Interferon-gamma; Interleukin-2; Interleukin-4; Male; Middle Aged; Multiple Sclerosis; Myelin Proteolipid Protein; Receptors, Antigen, T-Cell, alpha-beta; RNA, Messenger; T-Lymphocytes; Transforming Growth Factor beta

We studied glial transforming growth factor (TGF)-beta isotype expression in 14 cases of multiple sclerosis. Acute active lesions exhibited selective TGF-beta 2 immunoreactivity of lesion encircling ramified microglia. In contrast, astrocytes within chronic active white matter lesions expressed all three isotypes. Chronic active lesions which extended into cortex exhibited selective cortical astrocyte TGF-beta 2 expression. This isotype was also selectively expressed by astrocytes in apparently normal white matter. A similar pattern of glial TGF-beta expression was seen in the pathological control, progressive multifocal leukoencephalopathy. The results suggest that TGF-beta cytokines are locally expressed in demyelination and that the beta 2 isotype may be uniquely regulated.

Topics: Humans; Immunoenzyme Techniques; Multiple Sclerosis; Transforming Growth Factor beta

Multiple sclerosis (MS) is associated with high levels of circulating T lymphocytes that respond to the myelin antigens myelin basic protein (MBP) and proteolipid protein (PLP) by producing various cytokines including interferon-gamma (IFN-gamma) that makes MS worse and transforming growth factor-beta (TGF-beta), an endogenously produced immunosuppressant that might act beneficially. To further define the role of TGF-beta in MS, we examined the effects of recombinant TGF-beta 1 (rTGF-beta 1) on autoantigen-mediated regulation of cytokines in MS and myasthenia gravis (MG). Blood mononuclear cells (MNC) were cultivated with or without rTGF-beta 1, and with or without autoantigen or the recall antigen PPD. MNC expressing cytokine mRNA were detected after in situ hybridization with radiolabeled cDNA oligonucleotide probes. Femtogram concentrations of rTGF-beta 1 suppressed MBP-, PLP- and PPD-induced upregulation of IFN-gamma, IL-4, IL-6, tumor necrosis factor-alpha (TNF-alpha), TNF-beta and perforin in MS, and acetylcholine receptor (AChR)-induced augmentation of these pro-inflammatory cytokines in MG, but had no effects on autoantigen- or PPD-induced expression of IL-10 or TGF-beta itself. rTGF-beta 1 also suppressed numbers of myelin antigen-reactive IFN-gamma- and IL-4-secreting cells in MS and AChR-reactive IFN-gamma and IL-4 secreting cells in MG. The selective suppressive effects of TGF-beta 1 on autoantigen-induced upregulation of pro-inflammatory cytokines makes TGF-beta 1 attractive as a treatment alternative in MS and MG.

Topics: Adult; Aged; Autoantigens; Cells, Cultured; Cytokines; Female; Gene Expression; Humans; In Situ Hybridization; Inflammation; Interferon-gamma; Interleukin-10; Interleukin-4; Leukocytes, Mononuclear; Male; Middle Aged; Multiple Sclerosis; Myasthenia Gravis; Myelin Basic Protein; Oligonucleotide Probes; Recombinant Proteins; RNA, Messenger; Transforming Growth Factor beta

The bone morphogenetic proteins (BMPs) constitute a novel subfamily of the transforming growth factor type beta (TGF-beta) supergene family. Here we demonstrate, using polymerase chain reaction (PCR) BMP-4 and BMP-5 messages in RNA isolated from multiple sclerosis (MS) plaque tissue. This is the first demonstration of BMP expression in an inflammatory lesion in general, and in MS in particular. However, BMP-4 and BMP-5 messages could be detected in RNA isolated from a Morbus Creutzfeldt-Jakob (CJD) lesion. Even in normal brain, RNA expression of BMP-4, but not that of BMP-5, was detected. Therefore, BMP-5 gene expression seems to be associated with MS and CJD lesions, whereas the BMP-4 gene appears to be constitutively expressed in the human brain. As TGF-beta s and BMPs are regulators of regenerative processes and contribute to regulation of chemoattraction and local immunoreactivity, BMP-4 and BMP-5 might be involved in aspects of MS lesion formation unknown so far. PCR analysis of human cell lines demonstrate BMP-4 and BMP-5 expression in leukocytic cells, suggesting that infiltrating leukocytes contribute at least in part to BMP-4 and BMP-5 mRNAs of the MS plaque.

Topics: Creutzfeldt-Jakob Syndrome; Gene Expression; Humans; Middle Aged; Multiple Sclerosis; Polymerase Chain Reaction; Proteins; RNA; Transforming Growth Factor beta

We determined the cytokine messenger RNA (mRNA) expression pattern of blood mononuclear cells in 29 patients with relapsing-remitting multiple sclerosis every 4 weeks over a period of 12 months. During this period 27 relapses occurred in 14 patients (48%). Progression of disease activity as assessed by the occurrence of new lesions on nonenhancing T2-weighted magnetic resonance images of the head was detected in 12 (48%) of 25 patients. Using a semiquantitative polymerase chain reaction we demonstrated significant increases in tumor necrosis factor-alpha mRNA expression in peripheral blood mononuclear cells prior to a relapse. In 24 (85%) of 27 relapses increased tumor necrosis factor-alpha mRNA expression preceded clinical symptoms by 4 weeks. A similar pattern was observed for lymphotoxin mRNA expression. At the same time, transforming growth factor-beta and interleukin-10 mRNA levels declined. Fluctuations in the mRNA expression of tumor necrosis factor-alpha were also observed in 6 patients with stable disease who had active magnetic resonance scans on follow-up. No correlation of disease activity was observed with interleukin-1 beta, -4, or -6, inferferon gamma or endothelin-1 mRNA expression. From these data it can be concluded that variations in cytokine mRNA expression in blood mononuclear cells are correlated with disease activity in relapsing-remitting multiple sclerosis. It may be a valuable parameter to monitor the immunological status of patients in future clinical trials.

Topics: Adult; Base Sequence; Female; Gene Expression Regulation; Humans; Interleukin-10; Leukocytes, Mononuclear; Male; Middle Aged; Molecular Sequence Data; Multiple Sclerosis; Polymerase Chain Reaction; Prospective Studies; Recurrence; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Multiple sclerosis (MS) is associated with upregulation of both proinflammatory (interferon-gamma, IFN-gamma) and immunosuppressive (transforming growth factor-beta, TGF-beta) cytokines. To examine a possible relation between the MS-related HLA haplotype Dw2 and cytokine profiles, we used in situ hybridization with labeled cDNA oligonucleotide probes to detect transcripts of the T helper type 1 (Th 1) cell related IFN-gamma, the Th2 cell related interleukin-4 (IL-4) and of TGF-beta in blood and cerebrospinal fluid (CSF) mononuclear cells from 62 patients with MS. Compared to patients with other neurological diseases and healthy controls, MS patients had elevated numbers of IFN-gamma, IL-4 and TGF-beta mRNA expressing cells in blood and further augmented in CSF. Although several HLA-Dw2-positive individuals showed very high numbers of cells expressing these cytokines, no significant difference was found in comparison with Dw2-negative patients. However, expression of IL-4 and TGF beta mRNA was significantly increased in patients with shorter duration and minor disability and, for IL-4, in patients still in the relapsing-remitting phase compared to patients with secondary chronic progressive MS. Surprisingly, these changes which favour a beneficial, disease-downregulating effect of IL-4 and TGF-beta in MS, were found to be confined to HLA-Dw2-positive patients. Our findings suggest that the HLA phenotype does not influence the overall level of immune reactivity in MS, but may distinguish subgroups characterized by particular cytokine expression patterns.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Cytokines; Disease Progression; Female; Gene Expression; HLA-D Antigens; Humans; In Situ Hybridization; Interferon-gamma; Interleukin-4; Leukocytes, Mononuclear; Male; Middle Aged; Multiple Sclerosis; Myelin Sheath; Nervous System Diseases; Phenotype; RNA, Messenger; Transforming Growth Factor beta

Studies on patients with monosymptomatic optic neuritis (ON) should give opportunities to identify features typical for early multiple sclerosis (MS). There are increased T and B cell responses to the myelin components myelin basic protein (MBP) and proteolipid protein (PLP) in both ON and MS, but there is little information on the types of cytokines produced by such cells. We describe the use of in situ hybridization with complementary DNA oligonucleotide probes to detect and enumerate mononuclear cells expressing mRNA for the cytokines interferon-gamma (IFN-gamma) which augments cell-mediated immunity; interleukin-4 (IL-4) which promotes the B cell response; and transforming growth factor beta (TGF-beta) that in many cases downregulates immune responses. Expression of these cytokines was studied in mononuclear cells from peripheral blood and cerebrospinal fluid (CSF) from patients with ON and MS after in vitro exposure to MBP and PLP, and in absence of antigen. There were elevated levels of cells that in response to MBP and PLP expressed IFN-gamma, IL-4 and TGF-beta mRNA in blood and further enriched in CSF in both ON and MS, compared to patients with other neurological diseases. The results suggest that IFN-gamma, IL-4 as well as TGF-beta are involved in both ON and MS, and that the cytokine profile in early MS as reflected by ON is not different from that in clinically definite MS.

Topics: Adult; Female; Humans; Interferon-gamma; Interleukin-4; Male; Middle Aged; Multiple Sclerosis; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Optic Neuritis; RNA, Messenger; Transforming Growth Factor beta

Multiple sclerosis (MS) is characterized by patchy accumulations of inflammatory cells combined with demyelination. There are mononuclear cells in blood and cerebrospinal fluid of patients with MS that produce interferon-gamma and interleukin-4 in response to myelin basic protein (MBP) and proteolipid protein (PLP). Here we describe autoantigen-induced production of transforming growth factor-beta (TGF-beta). This multifunctional cytokine has inhibitory effects on the growth, differentiation, and effector functions of activated T cells. Blood and cerebrospinal fluid cells were exposed in short-term cultures to MBP and PLP and, after hybridization with complementary DNA oligonucleotide probes, they were evaluated for TGF-beta mRNA expression. Patients with MS had higher numbers of MBP- and PLP-responsive TGF-beta mRNA expressing cells in blood compared with control patients with other neurological diseases or myasthenia gravis and a five- and threefold further increment in their cerebrospinal fluid. In blood of patients with myasthenia gravis, where the acetylcholine receptor (AChR) is a target for autoaggressive immunity, there were increased levels of AChR-responsive TGF-beta mRNA expressing cells. Thymectomized myasthenia gravis patients showed higher levels of TGF-beta mRNA expressing cells compared with patients not thymectomized. Numbers of cells responding to AChR in MS and MBP in myasthenia gravis did not differ from numbers found in absence of antigen. Patients with other neurological diseases showed infrequent and low responses to MBP, PLP, and AChR. Diseases with presumed autoimmune pathogenesis are associated with organ-specific autoantigen-induced TGF-beta production, which is increased after thymectomy.

Topics: Adult; Autoantigens; Cerebrospinal Fluid; Female; Gene Expression; Humans; Male; Middle Aged; Multiple Sclerosis; Myasthenia Gravis; Myelin Proteins; Myelin Proteolipid Protein; Receptors, Cholinergic; RNA, Messenger; Transforming Growth Factor beta

Multiple sclerosis (MS) is characterized by perivascular inflammation and high levels of circulating T and B lymphocytes that respond to the myelin antigens myelin basic protein (MBP) and proteolipid protein (PLP), thereby suggesting a role for immunoregulatory cytokines.. Blood mononuclear cells (MNC) were prepared from patients with MS, optic neuritis (ON), myasthenia gravis (MG), other inflammatory (OIND) and non-inflammatory neurological diseases (OND), and from patients with HIV infection and healthy controls. MNC expressing cytokine mRNA were detected by in situ hybridization with radiolabelled cDNA oligonucleotide probes. Numbers of cytokine mRNA expressing cells were presented per standard numbers of MNC.. MS patients had elevated numbers of MNC in blood expressing T helper type 1 (Th1) cell related interferon-gamma (IFN-gamma), Th2 cell associated interleukin-4 (IL-4) and the endogenously produced immunosuppressant transforming growth factor-beta (TGF-beta). IFN-gamma and TGF-beta correlated with MS disability: EDSS score < 3 was associated with high numbers of TGF-beta mRNA positive cells while IFN-gamma mRNA positive cells tended to be low. The reverse was seen in patients with EDSS > or = 3. Cultures of MNC in presence and absence of antigen revealed that MBP and PLP induced strong responses in MS reflected by high levels of IFN-gamma, IL-4 and TGF-beta mRNA expressing cells. Recombinant (r) TGF-beta 1 dose-dependently suppressed MBP-induced upregulation of the proinflammatory cytokines IFN-gamma, IL-4, IL-6, tumor necrosis factor-alpha, (TNF-alpha), TNF-beta and perforin, but not of the immunosuppressive and probably advantageous IL-10. Cytokine mRNA expressing cells were enriched in the MS patients' cerebrospinal fluid, as were the cytokine mRNA positive cells detected after culture in presence of MBP and PLP, reflecting an autonomy of the immune response in this compartment. ON, in many instances representing early MS, did not differ from clinically definite MS regarding profiles of IFN-gamma, IL-4 and TGF-beta. Also patients with MG had elevated numbers of IFN-gamma, IL-4 and TGF-beta mRNA expressing blood MNC. They were further augmented upon culture of the MG patients' MNC in presence of acetylcholine receptor (AChR). An upregulation of AChR-induced TGF-beta was observed in thymectomized patients. rTGF-beta suppressed AChR-induced upregulation of proinflammatory cytokines but not IL-10. Elevated numbers of IFN-gamma, IL-4 and TGF-beta mRNA expressing blood MNC were also found in patients with OIND (aseptic meningo-encephalitis, chronic inflammatory demyelinating polyneuropathy, polymyositis, Eaton-Lambert syndrome) and in HIV-infected patients. In HIV infection, numbers of IL-4 mRNA positive cells correlated inversely with CD4+ cell counts, reflecting the involvement of IL-4 in later stages of the disease. Patients with non-inflammatory neurological diseases and healthy subjects had either no or low numbers of IFN-gamma, IL-4 and TGF-beta mRNA expressing cells when blood MNC were examined without previous culture, and after culture in presence and absence of MBP, PLP and AChR as antigens. An exception was a healthy pregnant lady who showed high le. High numbers of in vivo activated and of organ-specific antigen-responsive Th1 and Th2 like cells expressing IFN-gamma and IL-4 mRNA are characteristic for MS and MG. Upregulation of TGF-beta in MS patients with little disability and in MG after thymectomy implicates that TGF-beta has desirable effects in human diseases with autoimmune background.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; DNA, Complementary; Female; Gene Expression Regulation; HIV Infections; Humans; Interferon-gamma; Interleukin-4; Lambert-Eaton Myasthenic Syndrome; Male; Middle Aged; Monocytes; Multiple Sclerosis; Myasthenia Gravis; Oligonucleotide Probes; Optic Neuritis; Polymyositis; Polyradiculoneuropathy; RNA, Messenger; Transforming Growth Factor beta

Using a human culture system, we have previously shown that interferon-gamma-and tumour necrosis factor-alpha-stimulated astrocytes are capable of presenting antigens to T lymphocytes, but do not support antigen-dependent T cell proliferation. To gain further insight into the mechanisms involved in the local regulation of intracerebral T cell responses, we have investigated the effects of astrocytes on T cell proliferation induced by peripheral blood-derived mononuclear cells (PBMC). We found that astrocytes derived from human embryonic brain were able to suppress PBMC-dependent proliferation of antigen-specific, CD4+ T cell lines. Interferon-gamma production by PBMC-stimulated T cells was also suppressed by astrocytes, and this inhibition was seen as early as 6 h after initiation of co-culture. The inhibitory effect was observed in the presence of both HLA matched and mismatched astrocytes and was mediated by astrocyte-derived soluble factor(s) rather than by direct cellular contact. Inhibition of T cell proliferation was incompletely reverted by indomethacin, suggesting that prostaglandins were partially involved in the suppressive effect. The cytotoxic mediator nitric oxide was not involved in astrocyte-mediated inhibition. These observations led us to further investigate the contribution of other mediators known to down-regulate inflammatory processes. Our astrocyte cultures did not synthesize interleukin (IL)-4 or IL-10, whereas they secreted both the latent and active forms of transforming growth factor-beta 2. Transforming growth factor-beta was, however, found not to participate in astrocyte-induced inhibition in vitro. The inhibitory properties of human astrocytes may contribute to confinement of inflammatory lesions in multiple sclerosis and other inflammatory diseases of the central nervous system.

Topics: Astrocytes; Brain; Cell Division; Embryo, Mammalian; Humans; Interferon-gamma; Interleukin-4; Multiple Sclerosis; T-Lymphocytes; Transforming Growth Factor beta

The inflammatory nature of multiple sclerosis (MS) implicates the participation of immunoregulatory cytokines, including the T-helper type 1 (Th1) cell-associated interferon-gamma (IFN-gamma), the Th2 cell-related interleukin-4 (IL-4), and the immune response-downregulating cytokine transforming growth factor-beta (TGF-beta), but proof for their involvement in MS has been lacking. By adopting in situ hybridization with complementary DNA oligonucleotide probes for human IFN-gamma IL-4, and TGF-beta, the expression of mRNA for these cytokines was detected in mononuclear cells (MNC) from blood and cerebrospinal fluids. Strongly elevated levels of MNC expressing all three cytokines were found in peripheral blood and at even higher frequencies in cerebrospinal fluid from untreated patients with MS and optic neuritis, i.e., a common first manifestation of MS, compared with patients with other neurological diseases and healthy subjects. In MS and optic neuritis, IL-4 mRNA expressing cells predominated, followed by TGF-beta- and IFN-gamma-positive cells. Control patients with myasthenia gravis had similarly elevated levels of IFN-gamma and TGF-beta and TGF-beta mRNA expressing blood MNC but lower numbers of IL-4-positive cells. No or slight disability of MS was associated with high levels of TGF-beta mRNA expressing cells, while MS patients with moderate or severe disability had high levels of IFN-gamma-positive cells. IFN-gamma and TGF-beta may have opposing effects in MS, and treatments inhibiting IFN-gamma and/or promoting TGF-beta might ameliorate MS.

Topics: Adult; Female; Humans; In Situ Hybridization; Interferon-gamma; Interleukin-4; Male; Middle Aged; Multiple Sclerosis; RNA, Messenger; Transforming Growth Factor beta

Many chronic diseases follow a course with multiple relapses into periods with severe symptoms alternating with periods of remission; experimental allergic encephalomyelitis, the animal model for multiple sclerosis, is an example of such a disease. A finite Markov chain is proposed as a model for analyzing sequences of ordinal data from a relapsing-remitting disease. The proposed model is one in which the state space is expanded to include information about the relapsing-remitting status as well as the ordinal severity score, and a reparameterization is suggested that reduces the number of parameters needed to be estimated. The Markov model allows for a wide range of relapsing-remitting behavior, provides an understanding of the stochastic nature of the disease process, and allows for efficient estimation of important characteristics of the disease course (such as mean first passage times, occupation times, and steady-state probabilities). These methods are applied to data from a study of the effect of a treatment (transforming growth factor-beta 1) on experimental allergic encephalomyelitis.

Topics: Animals; Biometry; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Humans; Markov Chains; Mice; Models, Statistical; Multiple Sclerosis; Transforming Growth Factor beta

Activated T lymphocytes play an important role in the pathogenesis of multiple sclerosis (MS). These T cells secrete both pro- and anti-inflammatory cytokines. We have studied the production of these two kinds of cytokines by PBL of patients with MS and compared it with normal controls and other autoimmune diseases (OAD). PBL of 29 patients with MS, 14 patients with OAD, and 14 healthy normal controls were cultured for 5 wk. PBL of MS patients produced more pro-inflammatory cytokines, IL-2, IFN-gamma and TNF/lymphotoxin, and less anti-inflammatory cytokine, TGF-beta, during wk 2 to 4 in culture than PBL of normal controls. PBL of MS patients also produced more IL-2 and TNF/lymphotoxin than PBL of OAD patients. Decreased TGF-beta production by lymphocytes of patients with MS correlated directly with disease activity. MS patients with active disease produced less TGF-beta than MS patients with stable disease. The cells producing TGF-beta were primarily CD8+ T cells and CD45RA+T cells. These findings emphasize the complexity of immune response in MS patients and suggest that the increased production of pro-inflammatory cytokines by lymphocytes of patients with MS, combined with the decreased production of TGF-beta (anti-inflammatory cytokine), may play an important role in the mechanisms and manifestations of MS.

Topics: Adult; Autoimmune Diseases; Cell Line; Female; Humans; Interferon-gamma; Interleukin-2; Interleukin-4; Lymphocyte Activation; Male; Middle Aged; Multiple Sclerosis; T-Lymphocyte Subsets; T-Lymphocytes; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Samples representative of different stages of disease from a longitudinal study of multiple sclerosis patients were tested in the anchorage-independent growth assay for TGF-beta and an increased activity was detected in the supernatants from 2-day blood cell cultures from patients with active disease compared to patients without active disease and healthy donors. Within the group of patients with active disease, the TGF-beta like activity was significantly increased in the subgroup of patients tested during the period of regression of the symptoms where it appeared in 86.9% of the samples. These results suggest that TGF-beta or a TGF-beta like factor may play a role in regeneration processes in multiple sclerosis.

Topics: Adrenal Cortex Hormones; Adrenocorticotropic Hormone; Adult; Azathioprine; Female; Follow-Up Studies; Humans; Male; Middle Aged; Multiple Sclerosis; Neurologic Examination; Transforming Growth Factor beta

Patients with chronic progressive multiple sclerosis (MS) have alterations of T cell regulation that can be measured by in vitro assays and include decreases of the autologous mixed lymphocyte reaction (AMLR). Whether a defect in cytokine secretion was involved in the altered AMLR was investigated in 29 MS patients and 13 age- and sex-matched controls. The response of CD4+ T cell populations to irradiated non-T cells was decreased in MS as compared to control subjects. As previously reported, decreases in the AMLR were similarly observed with whole T cells of MS subjects as compared to controls. The addition of recombinant interleukin (IL)-1 to cultures of either whole T cells or CD4+ T cells with irradiated non-T cells in the AMLR corrected the immune defect in subjects with MS but had no effect on the AMLR in control subjects. In contrast, addition of rIL-2 or rIL-4 to the AMLR did not correct the decreased AMLR in MS patients as compared to controls. The lymphokines IFN-gamma and TGF-beta 2 both decreased the AMLR in MS patients and controls while TNF had no effect. Further, the magnitude of the AMLR response corresponded to IL-1 secretion induced by LPS in the non-T cell population. These studies indicate that defects in IL-1 may be related to immune defects of suppression in MS patients. Selective correction of immunoregulatory defects using lymphokines or their inducers in subjects with autoimmune diseases such as MS may be possible.

Topics: CD4-Positive T-Lymphocytes; Cytokines; Female; Humans; Interferon-gamma; Interleukin-1; Interleukin-2; Interleukin-4; Lymphocyte Culture Test, Mixed; Lymphocyte Depletion; Male; Multiple Sclerosis; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Topics: Brain; Gene Expression Regulation; Humans; Multiple Sclerosis; Promoter Regions, Genetic; Transforming Growth Factor beta

Blood macrophages and brain macrophages (microglia) have been implicated in demyelination and destruction of the oligodendrocyte in multiple sclerosis (MS), a disease affecting primarily white matter of the central nervous system (CNS). In this study, we demonstrate that at high effector to target cell ratios, normal rat microglia exhibit a natural cytotoxicity against normal rat oligodendrocytes in vitro. The killing is not mediated by the release of soluble factors. The cytotoxic activity is upregulated by pretreatment of microglia with interferon gamma (IFN gamma) or phorbol myristate acetate (PMA). Both the natural and induced cytotoxicities are inhibitable by transforming growth factor beta (TGF beta). The increase in numbers and apposition of primed or activated microglia to oligodendrocytes in MS lesions may give rise to natural or induced killing from which oligodendrocytes may be protected by TGF beta.

Topics: Animals; Brain; Cells, Cultured; Cytotoxicity, Immunologic; Depression, Chemical; Free Radicals; Interferon-gamma; Interleukin-1; Macrophage Activation; Macrophages; Multiple Sclerosis; Oligodendroglia; Rats; Tetradecanoylphorbol Acetate; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha