Proteomic analysis of cerebrospinal fluid from multiple sclerosis patients

Multiple sclerosis is an autoimmune inflammatory demyelinating disease of the central nervous system. Disease mechanisms in multiple sclerosis at the molecular level remain poorly understood and no reliable proteinaceous disease markers are available yet. The goal of the present study is the construction of a protein database of two-dimensional gel electrophoresis (2-DE) separated cerebrospinal fluid (CSF) proteins from multiple sclerosis patients. By means of liquid chromatography tandem mass spectrometry 65 different proteins were identified from 300 spots. Eighteen of these proteins have not been reported previously on 2-DE gels of CSF. Here we report on the identification of these proteins and discuss their potential relation to multiple sclerosis.     

Effects of Ferrofluid and Phytoalexin Spirobrassinin on Thioflavin-T-Based Fluorescence in Cerebrospinal Fluid of the Elderly and Multiple Sclerosis Patients

It is well known that misfolded peptides/proteins can play a role in processes of normal ageing and in the pathogenesis of many diseases including Alzheimer’s disease. Previously, we evaluated samples of cerebrospinal fluid from patients with Alzheimer’s disease and multiple sclerosis by means of thioflavin-T-based fluorescence. We observed attenuated effects of magnetite nanoparticles operated via anti-aggregation actions on peptides/proteins from patients with Alzheimer’s disease but not from those with multiple sclerosis when compared to age-related controls. In this study, we have evaluated the in vitro effects of anti-aggregation operating ferrofluid and phytoalexin spirobrassinin in the cerebrospinal fluid of patients with multiple sclerosis and Alzheimer’s disease. We have found significant differences in native fluorescence (λ excitation = 440 nm, λ emission = 485 nm) of samples among particular groups (young controls < multiple sclerosis, Alzheimer’s disease < old controls). Differences among groups were observed also in thioflavin-T-based fluorescence (young controls = multiple sclerosis < Alzheimer’s disease < old controls) and the most marked change from native to thioflavin-T-based fluorescence was found in young controls (28–40 years old people). Both ferrofluid and spirobrassinin evoked drops in thioflavin-T-based fluorescence; however, ferrofluid was more efficient in old controls (54–75 years old people) and spirobrassinin in multiple sclerosis patients, both compared to young controls. The results are discussed especially in relation to aggregated peptides/proteins and liposoluble fluorescent products of lipid peroxidation. Based on the significant effect of spirobrassinin in vitro, we suggest that spirobrassinin may be of therapeutic value in multiple sclerosis.     

Magnetic resonance imaging of the brain in multiple sclerosis

The paper deals with the diagnosis of demyelinating diseases of the central nervous system, including multiple sclerosis, by means of magnetic resonance imaging (MRI). The low-magnetic induction tomographs "Obraz" ("Image") and "Ellips" ("Ellipse") made in Russia were used to perform MRI of the brain in 255 patients diagnosed as having multiple sclerosis. Whether low-magnetic induction MRI can detectfocal changes in the brain in the presence of multiple sclerosis was assessed; the tomographic signs of the disease were clarified and classified. There was a relationship between the detection of focal changes in the brain and the duration of the disease. Particular emphasis is laid on the cases of various abnormalities at MRI, which clinically mask multiple sclerosis. The cases are summarized and a list of conditions to be particularly emphasized while making a differential diagnosis is drawn up.     

Pretreatment of Astrocytes with Interferon-α/β Prevents Neuronal Mitochondrial Respiratory Chain Damage

Abstract: Excessive nitric oxide/peroxynitrite generation has been implicated in the pathogenesis of multiple sclerosis, and the demonstration of increased astrocytic nitric oxide synthase activity in the postmortem brain of multiple sclerosis patients supports this hypothesis. Interferon-β is used for the treatment of multiple sclerosis, but currently little is known regarding its mode of action. Exposure of astrocytes in culture to interferon-γ plus lipopolysaccharide results in stimulation of nitric oxide release. Using a coculture system, we have been able to use astrocytes as a source of nitric oxide/peroxynitrite in an attempt to "model" the effects of raised cytokine levels observed in multiple sclerosis and to monitor the effect on neurones. Our results indicate that stimulation of astrocytic nitric oxide synthase activity causes significant damage to the mitochondrial activities of complexes II/III and IV of neighbouring neurones. This damage was prevented by a nitric oxide synthase inhibitor, suggesting that the damage was nitric oxide-mediated. Furthermore, interferon-α/β also prevented this damage. In view of these results, we suggest that a possible mechanism of action of interferon-β in the treatment of multiple sclerosis is that it prevents astrocytic nitric oxide production, thereby limiting damage to neighbouring cells, such as neurones.     

Epstein-Barr virus stimulates torque teno virus replication: a possible relationship to multiple sclerosis

Viral infections have been implicated in the pathogenesis of multiple sclerosis. Epstein-Barr virus (EBV) has frequently been investigated as a possible candidate and torque teno virus (TTV) has also been discussed in this context. Nevertheless, mechanistic aspects remain unresolved. We report viral replication, as measured by genome amplification, as well as quantitative PCR of two TTV-HD14 isolates isolated from multiple sclerosis brain in a series of EBV-positive and -negative lymphoblastoid and Burkitt's lymphoma cell lines. Our results demonstrate the replication of both transfected TTV genomes up to day 21 post transfection in all the evaluated cell lines. Quantitative amplification indicates statistically significant enhanced TTV replication in the EBV-positive cell lines, including the EBV-converted BJAB line, in comparison to the EBV-negative Burkitt's lymphoma cell line BJAB. This suggests a helper effect of EBV infections in the replication of TTV. The present study provides information on a possible interaction of EBV and TTV in the etiology and progression of multiple sclerosis.     

Estrogen receptor transrepresses brain inflammation

Estrogen receptors (ERs) have long been implicated in the etiology of multiple sclerosis, but no clear molecular mechanisms have linked ERs to the disease's pathology. Now Saijo et?al. (2011) provide evidence that ERβ activates a transrepression pathway that suppresses inflammation and inhibits progression of pathology in a mouse model of multiple sclerosis.     

Persistent or Slow Viral Infections and Related Diseases

The discovery of persistent transmissible agents by veterinarians has led to striking advances in the infectious cause of neuropathies of human beings. There is evidence for persisting infection in congenital rubella and the herpes group of viruses including cytomegalovirus infections. Hepatitis types A and B are candidates for inclusion in the category of persisting viral infections.The rubeola or measles virus is established as a persistent virus which causes elevated antibodies in the serum and cerebrospinal fluid of many patients with severe demyelinating disease such as subacute sclerosing panencephalitis and multiple sclerosis. Elevated antibodies against vaccinia virus have been found in the cerebrospinal fluid of some patients with multiple sclerosis and neuromyelitis optica, a rare form of multiple sclerosis.     

Vitamin D and autoimmunity: is vitamin D status an environmental factor affecting autoimmune disease prevalence?

The environment in which the encounter of antigen with the immune system occurs determines whether tolerance, infectious immunity, or autoimmunity results. Geographical areas with low supplies of vitamin D (for example Scandinavia) correlate with regions with high incidences of multiple sclerosis, arthritis, and diabetes. The active form of vitamin D has been shown to suppress the development of autoimmunity in experimental animal models. Furthermore, vitamin D deficiency increases the severity of at least experimental autoimmune encephalomyelitis (mouse multiple sclerosis). Targets for vitamin D in the immune system have been identified, and the mechanisms of vitamin D-mediated immunoregulation are beginning to be understood. This review discusses the possibility that vitamin D status is an environmental factor, which by shaping the immune system affects the prevalence rate for autoimmune diseases such as multiple sclerosis, arthritis, and juvenile diabetes.     

New approaches to the therapy of demyelinating disease.

The pace of research on the pathogenesis and treatment of multiple sclerosis, the principal human demyelinating disease of the central nervous system, has intensified in the past 3 years, due in part, to the application of advances in molecular and cellular immunology. Many lessons that have been learned in an animal model of central nervous system demyelinating disease, experimental allergic encephalomyelitis, also apply to multiple sclerosis and certain successful approaches for the treatment of this disease are now being attempted in humans.     

A Passive Exoskeleton Can Push Your Life Up: Application on Multiple Sclerosis Patients

In the present study, we report the benefits of a passive and fully articulated exoskeleton on multiple sclerosis patients by means of behavioral and electrophysiological measures, paying particular attention to the prefrontal cortex activity. Multiple sclerosis is a neurological condition characterized by lesions of the myelin sheaths that encapsulate the neurons of the brain, spine and optic nerve, and it causes transient or progressive symptoms and impairments in gait and posture. Up to 50% of multiple sclerosis patients require walking aids and 10% are wheelchair-bound 15 years following the initial diagnosis. We tested the ability of a new orthosis, the “Human Body Posturizer”, designed to improve the structural and functional symmetry of the body through proprioception, in multiple sclerosis patients. We observed that a single Human Body Posturizer application improved mobility, ambulation and response accuracy, in all of the tested patients. Most importantly, we associated these clinical observations and behavioral effects to changes in brain activity, particularly in the prefrontal cortex.     

The appearance of pars planitis in multiple sclerosis

The aim of the study was to investigate the clinical association of multiple sclerosis and pars planitis (or intermediate uveitis), as well as to determine the incidence of pars planitis in multiple sclerosis patients. During the period of one year authors examined 42 patients with multiple sclerosis divided into two groups. First group consisted of 23 patients with history of optic neuritis and the second group consisted of 19 patients who have never had optic neuritis. The mean age of patients in the first group was 31.7 +/- 5.1 years and in the second group 29.1 +/- 8.1 years. Pars planitis was found in 12 patients with multiple sclerosis. Age, sex and degree of neurological disability had no influence on the appearance of pars planitis. Although optic neuritis is considered to be the most common ocular manifestation of multiple sclerosis, the significant number of patients with multiple sclerosis has pars planitis.     

Helminth therapy and multiple sclerosis

Multiple sclerosis is a common and frequently disabling neurological disease of young adults. It is characterised by recurrent areas of focal inflammation (plaques) in the CNS which give rise to episodic neurological signs and symptoms. According to the hygiene (microbial deprivation) hypothesis, evolutionarily abnormal high levels of sanitation in the environment of the developed world may contribute to disordered immunoregulation in this and other putative autoimmune disorders. Helminths have been shown to augment immunoregulation. On this basis, the possibility of treating multiple sclerosis with live helminths or helminth products has been explored in animal models, natural human infections and phase 1 clinical trials. To date helminth therapy appears safe and preliminary clinical, magnetic resonance imaging and immunological outcomes have generally been favourable. Nevertheless, serious adverse effects are always possible, particularly with live parasitic administration. Follow up studies with safety monitoring, regulatory oversight and objective outcome measures will be required to definitively assess safety and efficacy for this novel class of potential immunological therapies in multiple sclerosis.     

Prevalence of multiple sclerosis in five rural Suffolk practices.

OBJECTIVE--To determine a point prevalence of multiple sclerosis in part of Suffolk. DESIGN--Multiple source search for patients with multiple sclerosis in five general practices. Patients were reviewed and categorised by using general practice notes. SETTING--Five rural general practices in Suffolk, 12 May 1988. SUBJECTS--31,379 patients registered with five practices. MAIN OUTCOME MEASURES--Multiple sclerosis diagnosed by a specialist. RESULTS--The search produced a provisional list of 62 eligible patients with multiple sclerosis. Review of case notes showed that 48 had probable disease, 10 early disease, and four possible disease. The probable cases gave a crude prevalence of 153/100,000 population (95% confidence interval 109/100,000 to 196/100,000). CONCLUSIONS--Although the results should be interpreted cautiously because of the small sample size, they suggest that the prevalence of multiple sclerosis in Suffolk is higher than has been estimated from hospital data.     

Characterization of antigen-specific T cells in multiple sclerosis twins with elevated proliferative responses to measles virus

The proliferative response to measles virus in normal individuals is low compared with the response to mumps virus. This is probably due to a low precursor frequency of OKT4+, IL 2-secreting helper cells. The presence of a measles high-responder state has previously been identified in some twin individuals with multiple sclerosis. Further characterization of the measles response in these high-responder individuals has demonstrated that the enhanced measles responses are due to a greater response by OKT4+ cells, which secrete higher levels of IL 2; this contrasting with the low levels of IL 2 secretion and OKT4+ cell proliferation seen in the unaffected twins. No evidence for suppression by either accessory or T cells, which would account for the quantitative differences between the high responders with multiple sclerosis and their unaffected low-responder twin siblings, was detected. The results indicate that a clonally expanded population of measles-specific responder cells is responsible for the high-responder state in these twins with multiple sclerosis. The mechanism producing this state may have relevance to possible immunoregulatory abnormalities producing autoimmunity in multiple sclerosis.     

The etiology of multiple sclerosis: genetic evidence for the involvement of the human endogenous retrovirus HERV-Fc1

We have investigated the role of human endogenous retroviruses in multiple sclerosis by analyzing the DNA of patients and controls in 4 cohorts for associations between multiple sclerosis and polymorphisms near viral restriction genes or near endogenous retroviral loci with one or more intact or almost-intact genes. We found that SNPs in the gene TRIM5 were inversely correlated with disease. Conversely, SNPs around one retroviral locus, HERV-Fc1, showed a highly significant association with disease. The latter association was limited to a narrow region that contains no other known genes. We conclude that HERV-Fc1 and TRIM5 play a role in the etiology of multiple sclerosis. If these results are confirmed, they point to new modes of treatment for multiple sclerosis.     

Specific antibodies reacting to JC polyomavirus capsid protein mimotopes in sera from multiple sclerosis and other neurological diseases-affected patients

Published data support the hypothesis that viruses could be trigger agents of multiple sclerosis onset. This link is based on evidence of early exposure to viral agents in patients affected by this neurologic disease. JC (JC polyomavirus [JCPyV]), BK (BKPyV), and simian virus 40 (SV40) neurotropic polyomavirus footprints have been detected in brain tissue specimens and samples from patients affected by different neurological diseases. In this investigation, serum samples from patients affected by multiple sclerosis and other inflammatory and noninflammatory neurologic diseases, as well as healthy subjects representing the control, were investigated for immunoglobulin G (IgG) antibodies against JCPyV. To this end, an immunologic approach was employed, which consists of employing indirect enzyme-linked immunosorbent assay testing with synthetic peptides mimicking viral capsid protein 1 antigens. A significantly lower prevalence of IgG antibodies against JCPyV VP1 epitopes, with a low titer, was detected in serum samples from patients with multiple sclerosis (MS) and other neurologic diseases than in healthy subjects. Our study indicates that the prevalence of JCPyV antibodies from patients with multiple sclerosis is 50% lower than in healthy subjects, suggesting specific immune impairments. These results indicate that patients affected by neurological diseases, including MS, respond poorly to JCPyV VP1 antigens, suggesting specific immunologic dysfunctions.     

Differential Micro RNA Expression in PBMC from Multiple Sclerosis Patients

Differences in gene expression patterns have been documented not only in Multiple Sclerosis patients versus healthy controls but also in the relapse of the disease. Recently a new gene expression modulator has been identified: the microRNA or miRNA. The aim of this work is to analyze the possible role of miRNAs in multiple sclerosis, focusing on the relapse stage. We have analyzed the expression patterns of 364 miRNAs in PBMC obtained from multiple sclerosis patients in relapse status, in remission status and healthy controls. The expression patterns of the miRNAs with significantly different expression were validated in an independent set of samples. In order to determine the effect of the miRNAs, the expression of some predicted target genes of these were studied by qPCR. Gene interaction networks were constructed in order to obtain a co-expression and multivariate view of the experimental data. The data analysis and later validation reveal that two miRNAs (hsa-miR-18b and hsa-miR-599) may be relevant at the time of relapse and that another miRNA (hsa-miR-96) may be involved in remission. The genes targeted by hsa-miR-96 are involved in immunological pathways as Interleukin signaling and in other pathways as wnt signaling. This work highlights the importance of miRNA expression in the molecular mechanisms implicated in the disease. Moreover, the proposed involvement of these small molecules in multiple sclerosis opens up a new therapeutic approach to explore and highlight some candidate biomarker targets in MS.     

NMDA receptors, glial cells, and clinical medicine

Recent reports have overturned a series of dogmas that have been well entrenched in the neuroscience literature concerning NMDA-type glutamate receptors (NMDARs). The new data show that NMDARs exist on the myelin sheath formed by oligodendrocytes, that an uncompetitive NMDAR antagonist has successfully passed human clinical trials, and that NMDARs trigger multiple deleterious cascades to inflict cellular damage on both neurons and glia during cerebral ischemia (stroke). These recent findings bode well for clinical intervention with NMDAR antagonists in more neurological disorders than previously thought, including multiple sclerosis, cerebral palsy (periventricular leukomalacia), and spinal cord injury.     

A chromosome study in multiple sclerosis

Summary Chromosome studies of 30 patients with multiple sclerosis and 30 controls have been made. The results show that there is no significant increase in the frequency of structural aberrations in the multiple sclerosis patients. It is suggested that previously reported differences may have been due to technical factors, the nature of which is discussed.

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Zusammenfassung Bei 30 Patienten mit multipler Sklerose und 30 Kontrollen wurden die Chromosomen untersucht. Es fand sich bei den Patienten kein signifikanter Anstieg in der Häufigkeit struktureller Aberrationen. Vielleicht wurden früher mitgeteilte Unterschiede durch technische Faktoren, die im einzelnen diskutiert werden, vorgetäuscht.

     

Measles antigen in multiple sclerosis: Identification in the jejunum by immunofluorescence

Measles virus antigen has been detected by the fluorescent antibody method in jejunal mucosa obtained from 24 patients with clinical multiple sclerosis. Deposits of antigen, various components of the complement system and on occasion immunoglobulins were identified in the epithelial basement membrane. Antigen and complement were usually found in the lamina propria, and sometimes could be seen within epithelial cells and in capillary walls. These findings support the concept that multiple sclerosis is caused by persistent measles infection, and indicate that the virus is harbored in the wall of the small intestine.