Journal of Neuroimmunology - Articles in Press

Exploring the role of the epigenome in multiple sclerosis: A window onto cell-specific transcriptional potential - Corrected Proof

Anna Kaliszewska, Philip L. De Jager — 2012-02-02

Abstract: The field of epigenomics involves the study of chromatin, the three dimensional complex of DNA, protein and non-coding RNAs that determines the accessibility of DNA by the transcriptional machinery. The epigenome varies from cell to cell and reflects the effect of external stimuli on cell fate and cell state. Thanks to emerging platforms and analysis methods, the systematic characterization of chromatin conformation throughout the genome has begun and has yielded several reference epigenome maps for a growing number of cell types. Such maps are enabling insights into the correlation architecture of different epigenomic marks: a number of discrete chromatin states are found across different cell types. The combination of these reference maps and robust platforms for genome-wide data generation has introduced a new era in which studies of human disease are becoming feasible. Little is known about the role of the epigenome in MS, but it is likely that, as in other inflammatory disease, susceptibility factors and events along the course of the disease will alter the chromatin state of different cell types in patients with MS. Here, we review different strategies for the characterization of the epigenome and how these strategies could be used to implement new studies to explore how alterations of chromatin architecture establish a dysregulated transcriptional state in the context of MS.

Next-generation sequencing approaches for genetic mapping of complex diseases - Corrected Proof

Ferran Casals, Youssef Idaghdour, Julie Hussin, Philip Awadalla — 2012-01-30

Abstract: The advent of next generation sequencing technologies has opened new possibilities in the analysis of human disease. In this review we present the main next-generation sequencing technologies, with their major contributions and possible applications to the study of the genetic etiology of complex diseases.

Repeated morphine treatment-mediated hyperalgesia, allodynia and spinal glial activation are blocked by co-administration of a selective cannabinoid receptor type-2 agonist - Corrected Proof

2012-01-30

Abstract: Spinal glial activation has been implicated in sustained morphine-mediated paradoxical pain sensitization. Since activation of glial CB2 cannabinoid receptors attenuates spinal glial activation in neuropathies, we hypothesized that CB2 agonists may also attenuate sustained morphine-mediated spinal glial activation and pain sensitization. Our data indicate that co-administration of a CB2-selective agonist (AM 1241) attenuates morphine (intraperitoneal; twice daily; 6days)-mediated thermal hyperalgesia and tactile allodynia in rats. A CB2 (AM 630) but not a CB1 (AM 251) antagonist mitigated this effect. AM 1241 co-treatment also attenuated spinal astrocyte and microglial marker and pro-inflammatory mediator (IL-1β, TNFα) immunoreactivities in morphine-treated rats, suggesting that CB2 agonists may be useful to prevent the neuroinflammatory consequences of sustained morphine treatment.

Data integration and systems biology approaches for biomarker discovery: Challenges and opportunities for multiple sclerosis - Corrected Proof

Pablo Villoslada, Sergio Baranzini — 2012-01-27

Abstract: New “omic” technologies and their application to systems biology approaches offer new opportunities for biomarker discovery in complex disorders, including multiple sclerosis (MS). Recent studies using massive genotyping, DNA arrays, antibody arrays, proteomics, glycomics, and metabolomics from different tissues (blood, cerebrospinal fluid, brain) have identified many molecules associated with MS, defining both susceptibility and functional targets (e.g., biomarkers). Such discoveries involve many different levels in the complex organizational hierarchy of humans (DNA, RNA, protein, etc.), and integrating these datasets into a coherent model with regard to MS pathogenesis would be a significant step forward. Given the dynamic and heterogeneous nature of MS, validating biomarkers is mandatory. To develop accurate markers of disease prognosis or therapeutic response that are clinically useful, combining molecular, clinical, and imaging data is necessary. Such an integrative approach would pave the way towards better patient care and more effective clinical trials that test new therapies, thus bringing the paradigm of personalized medicine in MS one step closer.

Key role of CXCL13/CXCR5 axis for cerebrospinal fluid B cell recruitment in pediatric OMS - Corrected Proof

2012-01-24

Abstract: To study aberrant B cell trafficking into the CSF in opsoclonus–myoclonus syndrome (OMS), chemoattractants CXCL13 and CXCL12, and B cell frequency and CXCR5 expression, were evaluated. CSF CXCL13 concentration and the CSF/serum ratio were higher in untreated OMS than controls, related directly to OMS severity and inversely to OMS duration, and correlated with CSF B cell frequency and oligoclonal bands. CXCL12 showed the opposite pattern. Selective accumulation of CXCR5+ memory B cells in CSF was found. In ACTH-treated OMS, CXCL13, but not CXCL12, was lower. These data implicate the chemokine/chemoreceptor pair CXCL13/CXR5 in B cell recruitment to the CNS in OMS. CXCL13 and CXCL12 may serve as reciprocal biomarkers of disease activity, but CXCL13 also had utility as a treatment biomarker.

Elevation of serum TNF-α levels in mild and moderate Alzheimer patients with daytime sleepiness - Corrected Proof

2012-01-24

Abstract: Objectives: Sleep disturbance has been noted to accompany Alzheimer disease and is more pronounced as dementia severity increases. The aim of this study was to examine whether sleep disturbance in a cohort of patients with mild/moderate AD was associated with serum levels of IL-1β and TNF-α.Methods: Forty three drug-free AD patients and twenty two healthy controls were evaluated. All subjects underwent two consecutive full-night polysomnography. Their daytime sleepiness was assessed by Epworth Sleepiness Scale (ESS). Serum levels of IL-1β and TNF-α were measured by enzyme linked immunoassays.Results: AD patients showed lower sleep efficiency, more awakenings and less slow wave sleep (SWS). IL-1β was detectable only in two AD patients. Serum TNF-α concentrations did not differ significantly between AD patients and controls. When AD patients were classified as AD patients with daytime sleepiness (n=20, ESS>10) or AD patients without daytime sleepiness (n=23, ESS<10) according to their ESS scores, serum levels of TNF-α was significantly higher in AD patients with daytime sleepiness than that in those without daytime sleepiness or controls (32.7±17.9 vs 5.2±2.4, p<0.05; 40.9±22.3 vs 5.7±2.2, p<0.05). Serum level of TNF-α was significantly correlated with ESS score.Discussion: These data indicate that daytime sleepiness in mild and moderate AD patients is associated with elevation of serum TNF-α concentrations.

The CD4+ T-cell subset lacking expression of the CD28 costimulatory molecule is expanded and shows a higher activation state in multiple sclerosis - Corrected Proof

2012-01-20

Abstract: Multiple sclerosis (MS) is a chronic debilitating disease, in which T-cells are considered to play a pivotal role. CD28 is the quintessential costimulatory molecule on T-cells and its expression declines progressively with repeated stimulations, leading to the generation of CD28− T-cells. Our aim was to examine whether CD4+CD28− T-cells were enriched in MS patients, and characterize the phenotype of this subset in MS patients and healthy controls (HC).All these changes could provide these CD4+CD28− T-cell characteristics that might be involved in the pathogenesis of MS, turning this T-cell subset into a potential target for future therapeutic strategies.

Rapamycin reduces clinical signs and neuropathic pain in a chronic model of experimental autoimmune encephalomyelitis - Corrected Proof

2012-01-20

Abstract: Current treatments used in Multiple Sclerosis (MS) are partly effective in the early stages of the disease but display very limited benefits in patients affected by progressive MS. One possible explanation is that these therapies are unable to target the inflammatory component most active during the progressive phase of the disease, and compartmentalized behind the blood–brain barrier. Our findings show that Rapamycin ameliorates clinical and histological signs of chronic EAE when administered during ongoing disease. Moreover, Rapamycin significantly reduced the hyperalgesia observed before clinical development of EAE which, in turn, is completely abolished by the administration of the drug.

Identification of continuous epitopes of HuD antibodies related to paraneoplastic diseases/small cell lung cancer - Corrected Proof

2012-01-20

Abstract: HuD antibodies are associated with small cell lung cancer. To identify relevant epitopes of HuD antibodies, patient sera and a monoclonal antibody were analyzed for their reactivity to linear 20mer peptides spanning the human HuD protein. The HuD monoclonal antibody recognized a single fragment located in the first RNA recognition motif. Thorough analysis identified VRDKITQGSL as the actual epitope. Screening of anti-HuD positive patients and healthy controls identified eight peptides as potential subdominant epitopes. The majority of these peptides were located in the N-terminal end as well as in the linker region between the second and third RNA recognition motifs.

Meningothelial cells participate in immunological processes in the cerebrospinal fluid - Corrected Proof

2012-01-20

Abstract: Meningothelial cells (MECs) form the innermost layer of the meningeal sheath and as such are in direct contact with the cerebrospinal fluid (CSF) likely influencing CSF composition. The CSF space is a site of active immunological processes. To investigate an immunological role of MECs, cytokine and chemokine secretion, phagocytotic and pinocytotic activity by MECs was analyzed following stimulation with lipopolysaccharide, phorbol ester or rotenone. Secretion of IL-6 and IL-8 by MECs increased in a dose dependent manner after stimulation concomitant with NF-κB activation. In addition, phagocytotic clearance by MECs was enhanced suggesting an immunological role for MECs in the CSF compartment and pointing to a possible connection to neurodegenerative processes.

Associations of moderate alcohol consumption with clinical and MRI measures in multiple sclerosis - Corrected Proof

2012-01-19

Abstract: Objective: To examine the associations of alcohol consumption patterns with disability and brain injury in multiple sclerosis (MS) patients.Design: This study included 423 subjects (272 MS patients, 151 healthy controls) participating in a study of clinical, environmental and genetic risk factors in MS. Disability was assessed with the Expanded Disability Status Scale (EDSS) and the MS Severity Scale (MSSS). Brain injury was assessed using the quantitative MRI measures of T2-lesion volume (T2-LV), T1-LV, normalized volumes of brain parenchyma (NBV), gray matter (NGMV) and lateral ventricle (NLVV). Information related to alcohol-consumption patterns was obtained with standardized questionnaire during an in-person interview. The associations of alcohol consumption variables with disability and MRI measures were assessed in regression analyses.Results: The frequency of MS patients who did not consume alcohol after MS (19.4%) was higher than the frequency before MS (p<0.001). The EDSS, NGMV and NLVV exhibited a non-linear dependence on duration of alcohol consumption after MS onset: non-linear regression analyses indicated that EDSS and NLVV were lower and the NGMV was greater in MS patients who had consumed for a period of 15years or less after MS onset compared those who did not consume alcohol or consumed it for more than 15years.Conclusion: The duration of alcohol consumption is associated with disability and MRI measures in MS. Prospective, longitudinal studies of the role of alcohol in MS disease progression are warranted.

Frequency and function of regulatory T cells after ischaemic stroke in humans - Corrected Proof

2012-01-19

Abstract: Ischaemic stroke is an important cause of disability and death. Local inflammation in the brain following stroke is thought to enhance damage. Animal studies show that regulatory T cells (Tregs) can downregulate this inflammation and assist recovery, but there are no previous studies of the function of Tregs in human stroke. The current study aimed to quantify Tregs in peripheral blood following stroke and to investigate their function. Treg numbers were significantly increased after stroke, particularly in males. However, the functional capacity of Tregs to inhibit proliferation of autologous cells at low ratios of Treg to responder cells was reduced, particularly in female patients, compared to controls. This study is the first to report gender-specific changes in the numbers and function of Tregs after ischaemic stroke. These changes may affect stroke outcome.

The association of V249I and T280M fractalkine receptor haplotypes with disease course of multiple sclerosis - Corrected Proof

2012-01-19

Abstract: We investigated the association of CX3CR1 genotypes/haplotypes with MS and performed the prediction analysis of protein sequence variants' effects on CX3CL1/CX3CR1 interaction. We found no association of CX3CR1 with MS susceptibility. Frequency of I249T280 haplotype was significantly lower in SP compared to RR patients (RR>10years, OR=0.30, 95%CI=0.11–0.79, p=0.01; OR=0.53, 95%CI=0.18–1.56, p=0.2, in SP<10years vs. RR>10years). Prediction analysis showed that I249 T280 protein variant would significantly affect CX3CL1/CX3CR1 interaction. Our results suggest that CX3CR1 I249T280 haplotype could have protective effect for switch to SP MS. Further research is warranted to validate and replicate currently observed results.

Aberrant expression of the apoptosis-related proteins BAK and MCL1 in T cells in multiple sclerosis - Corrected Proof

Ilana Mandel, Tamar Paperna, Ariel Miller — 2012-01-18

Abstract: Pathogenic T cells of multiple sclerosis (MS) patients have been suggested to be endowed with an increased resistance to apoptosis, contributing to their increased survival. We report herein increased levels of the anti-apoptotic MCL1 protein and its half-life in activated lymphocytes of MS patients, which were not associated with differences in MCL1 RNA levels or with alterations in the expression levels of the known E3 ligases of MCL1-β-TrCP and HUWE1. Concomitantly, the expression levels of the pro-apoptotic protein BAK were decreased in MS patients at relapse. These findings suggest the dysregulation of the apoptosis-related proteins MCL1 and BAK in MS.

Upregulation of IL-17, but not of IL-9, in circulating cells of CIS and relapsing MS patients. Impact of corticosteroid therapy on the cytokine network - Corrected Proof

2012-01-16

Abstract: The concomitant production of IL-17A and IL-9, both Th17 cytokines, has not been compared in MS patients. We show that IL-17A but not IL-9 expression by CD3+ cells was increased during a MS relapse. Co-expression of IL-17A and IL-9 was marginal. In addition to Th1 and Th2 cytokines, IL-17A, IL-6 and IL-23p19 were down-regulated by ivMP, but Foxp3 was not, while an increase in IL-10, TGF-β1 and IL-27p28 mRNA was observed. This change in the Th17, Treg and IL-10 balance could be an additional mechanism by which corticosteroids shorten the duration of a MS relapse and promote recovery.

Mediatophore regulates acetylcholine release from T cells - Corrected Proof

Takeshi Fujii, Yuki Takada-Takatori, Kazuhide Horiguchi, Koichiro Kawashima — 2012-01-16

Abstract: Immunological stimulation of T cells by phytohemagglutinin (PHA) enhances the synthesis and release of acetylcholine (ACh), suggesting a role for the lymphocytic cholinergic system in the regulation of immune function. In the present study, we used two human leukemic T cell lines as models to investigate whether mediatophore, a homooligomer of a 16-kDa subunit homologous to the proteolipid subunit c of vacuolar H+-ATPase (V-ATPase), is involved in mediating ACh release from T cells. Immunohistochemical analysis revealed the presence of mediatophore in the cytoplasm and on the plasma membrane of both T cell lines. Mediatophore gene expression was up-regulated by immunological T cell activation by PHA. Transfection of anti-mediatophore small interference RNA down-regulated mediatophore gene expression and significantly reduced ACh release. These results suggest that T cells express mediatophore, which then plays a key role in mediating ACh release, and that mediatophore expression is regulated by immunological stimulation.

Analysis of antibodies to surface epitopes of contactin-2 in multiple sclerosis - Corrected Proof

2012-01-16

Abstract: Contactin-2 was recently identified as an autoantigen targeted by T-cells and autoantibodies in multiple sclerosis (MS). Here we analyzed the frequency of antibodies to contactin-2 (contactin-2-ab) by a cell-based assay in the serum from 105 MS patients and at least 5years of follow-up (19 clinically isolated syndromes, 51 relapsing–remitting, 20 secondary-progressive, and 15 primary-progressive). Contactin-2-ab were detected in 4 (7.8%) relapsing–remitting patients. Clinical and magnetic resonance imaging characteristics were not significantly different from the rest of relapsing–remitting patients. In conclusion, contactin-2-ab are identified in a minority of MS patients but their presence is not associated with a particular clinical-radiological profile.

Human T cell expansion and experimental autoimmune encephalomyelitis inhibited by Lenaldekar, a small molecule discovered in a zebrafish screen - Corrected Proof

2012-01-16

Abstract: Immune-mediated diseases [multiple sclerosis (MS), experimental autoimmune encephalomyelitis (EAE)] are driven by proliferating, highly activated autoreactive T-cells that are unresponsive to in vivo immunoregulatory mechanisms. The compound Lenaldekar (LDK) was identified in a zebrafish screen by inhibiting T-cell expansion. By monitoring mitogen- and antigen-driven proliferation, we found that LDK inhibited human and murine T-cell expansion in a non-cytolytic manner. This suppressive activity directly correlated with the degree of activation/proliferation of the T-cells. In testing LDK in an EAE model of MS, exacerbations were suppressed in treated animals. Therefore, LDK represents a novel therapeutic approach to T-cell-mediated autoimmune diseases.

Loss of class I MHC function alters behavior and stress reactivity - Corrected Proof

Ashwin Sankar, Robyn N. MacKenzie, Jane A. Foster — 2012-01-16

Abstract: The importance of the classical immune molecule, class I major histocompatibility complex to central nervous system function is one of the most surprising discoveries related to neuroimmunology in the past decade. Mice lacking both β-2microglobulin and transporter associated with antigen processing (β2M−/−TAP−/−) showed differences in basal behavior. In response to saline injection, β2M−/−TAP−/− mice showed a significant hypothalamic pituitary adrenal activation that was not observed in wild type mice, while lipopolysaccharide-induced cytokine expression in the hypothalamus was similar in β2M−/−TAP−/− and wild type mice. Overall, these data show that class I MHC plays an important role in behavior and stress reactivity.

No evidence for an anti-inflammatory effect of escitalopram intervention in healthy individuals with a family history of depression - Corrected Proof

Eva Haastrup, Ulla Knorr, Christian Erikstrup, Lars Vedel Kessing, Henrik Ullum — 2012-01-13

Abstract: Inflammation may contribute to the pathogenesis of depression and antidepressants are hypothesised to have an anti-inflammatory effect.In this randomised double-blinded trial we investigated the cytokine levels in supernatants of stimulated whole blood samples from first degree relatives to patients with depression randomised to a single daily dose of either 10mg escitalopram or placebo for four weeks.No significant differences were found in any of the cytokine levels between the participants treated with escitalopram (n=21) or placebo (n=23).Our data does thus not support the hypothesis of a global anti-inflammatory effect of escitalopram on cytokines in healthy subjects.

Stress and the anti-influenza immune response: Repeated social defeat augments clonal expansion of CD8+T cells during primary influenza A viral infection - Corrected Proof

2012-01-13

Abstract: Social disruption stress (SDR) prior to primary influenza A virus (IAV) infection augments memory to IAV re-challenge in a T cell-specific manner. However, the effect of SDR on the primary anti-viral immune response has not been elucidated. In this study, SDR-infected (INF) mice terminated viral gene expression earlier and mounted an enhanced pulmonary IAV-specific CD8+T cell response versus controls. Additionally, SDR-INF mice had a more pro-inflammatory lung profile prior to and during infection and an attenuated corticosterone response. These data demonstrate neuroendocrine modification of the lung microenvironment and increased antigen-specific T cell activation, clonal expansion and viral control in stress-exposed mice.

Palmitoylethanolamide stimulates phagocytosis of Escherichia coli K1 and Streptococcus pneumoniae R6 by microglial cells - Corrected Proof

Sandra Redlich, Sandra Ribes, Sandra Schütze, Dirk Czesnik, Roland Nau — 2012-01-13

Abstract: The ability of microglial cells to phagocytose bacteria after stimulation with the endocannabinoid palmitoylethanolamide (PEA) was studied in vitro. PEA increased the phagocytosis of unencapsulated Streptococcus pneumoniae R6 and encapsulated Escherichia coli K1 by murine microglial cells significantly after 30min of microglial stimulation. This suggested that stimulation of microglial cells by PEA can increase the resistance of the brain against CNS infections.

The effect of TNF-alpha, FcγR and CD1 polymorphisms on Guillain–Barré syndrome risk: Evidences from a Meta-Analysis - Corrected Proof

Li-ya Wu, You Zhou, Chao Qin, Bang-li Hu — 2012-01-12

Abstract: Background and objectivesThe findings on the associations between potential genetic variants and risk of Guillain–Barré syndrome (GBS) are controversial. We conducted a meta-analysis for candidate genes to provide the evidences for the current understanding of the genetic association with GBS.Methods: We searched relevant studies without language restriction in PubMed, Embase and Cochrane library through May 2011. The strengths of the associations between genetic variants and GBS risk were estimated by odds ratios (ORs) with 95% confidence intervals (CIs). Random-effects models or fixed effects model was applied based on the heterogeneity test.Results: We identified 12 case–control studies involving 1,590 GBS cases and 2,154 controls for the analysis. Because of limited eligible data, our meta-analysis specifically focused on 6 genetic variants of 3 candidate genes, TNF-α, FcγR and CD1. We found that TNF-α 308 G/A polymorphism was significantly associated with the risk of GBS in the overall population (GG+GA vs. AA: OR=0.32, 95%CI=0.16-0.62; GG vs. AA: OR=0.36, 95%CI=0.19-0.68). Subgroup analysis further provided evidence of significant association between TNF-α 308 G/A and risk of the GBS in Asian population (GG+GA vs. AA: OR=32, 95%CI=0.11-0.93; GG vs. AA: OR=0.32, 95%CI=0.15-0.68). In addition, we did not observe significant associations between FcγRIIA R/H, FcγRIIIA F/V, FcγRIIIB NA1/NA2, CD1A 1/2 and CD1E 1/2 polymorphisms and susceptibility for developing GBS.Conclusions: Our findings showed that TNF-α 308A allele might be a moderate risk factor for GBS. However, the results should be interpreted with caution due to the limited number of studies available.

Investigating channel blockers for the treatment of multiple sclerosis: Considerations with mefloquine and carbenoxolone - Corrected Proof

Remington Lee Nevin — 2012-01-11

I read with interest the recent article by describing inhibition of IL-23 expression by carbenoxolone. This article, together with earlier work demonstrating that carbenoxolone delayed onset of experimental autoimmune encephalomyelitis, plausibly through blockade of non-junctional pannexin channels or connexin hemichannels (), raises the intriguing possibility that channel blockers might be useful in the treatment of Th17-mediated autoimmune diseases, including multiple sclerosis (MS).

Serum levels of interleukin (IL)-18, IL-23 and IL-17 in Chinese patients with multiple sclerosis - Corrected Proof

2012-01-09

Abstract: It has been reported that cytokines play an important role in the pathogenesis of multiple sclerosis (MS). The aim of this study was to evaluate the serum levels of interleukin (IL)-18, IL-23 and IL-17 in Chinese patients with MS. We compared the serum concentrations of pro-inflammatory cytokines IL-18, IL-23 and IL-17 in 39 patients with MS and 39 healthy controls matched with sex and age. Serum cytokines were measured by FlowCytomix, a kind of cytometric bead-based assay. Correlations between the serum levels of the three cytokines and disability (expanded disability status scale, EDSS), disease duration, current age and age at onset were examined. Serum concentrations of all IL-18, IL-23 and IL-17 were significantly higher in MS patients than healthy controls. There were no significant differences of the three cytokines' levels between female and male healthy controls, while the serum IL-18 level was observed significantly higher (P=0.049) in male MS patients than female MS patients. No significant correlations were observed between any of the three cytokines' levels and EDSS, disease duration and current age. However, IL-23 was found negatively correlated with age at onset in male MS patients (rs=−0.775, P=0.041). Our data suggest that all IL-18, IL-23 and IL-17 may be involved in the pathogenesis of MS. However, the relationships of the three cytokines and clinical characteristics of MS need to be further investigated in the future.

Latent tuberculosis seems not to reactivate in multiple sclerosis patients on natalizumab - Corrected Proof

2012-01-09

Abstract: Current safety recommendations for multiple sclerosis (MS) patients who are considered for natalizumab do not specify how to screen for latent tuberculosis (LTB). Only chest X-ray is recommended as a routine, and follow-up depends on its results. The incidence of TB in Spain is high and the risk of a LTB reactivation due to natalizumab is unknown. Our objective is to describe in our clinical practice if following the current recommendations for the MS population on natalizumab allows identifying patients with a LTB, as well as the risk for TB reactivation. Our study demonstrated that, in our environment, current recommendations are not sensitive enough to identify cases of LTB, though no cases of active TB were observed. Considering the lack of documented active TB cases worldwide among natalizumab patients, we suggest that these safety measures are probably unnecessary, even in countries with a high TB incidence.

Divalent and monovalent autoantibodies cause dysfunction of MuSK by distinct mechanisms in a rabbit model of myasthenia gravis - Corrected Proof

2012-01-06

Abstract: Muscle-specific kinase (MuSK), a receptor tyrosine kinase, is required for the formation and maintenance of neuromuscular junctions (NMJs). Although autoantibodies against MuSK have been demonstrated to cause myasthenia gravis (MG), the underlying pathogenic mechanism remains unclear because a major subclass of these antibodies is functionally monovalent. We investigated the pathogenic role of MuSK antibodies in the onset of MG in vivo and in vitro. Ultrastructural visualization of NMJs in paretic rabbits with MuSK antibodies indicated that postsynaptic membranes were preserved, despite a significant loss of complexity in the convoluted synaptic folds. In addition, an in vitro assay indicated that both divalent and monovalent antibodies from paretic rabbits could interfere with agrin-induced acetylcholine receptor (AChR) clustering in cultured myotubes. Furthermore, in the absence of agrin, divalent antibodies induced MuSK phosphorylation and accelerated downregulation of Dok-7, an essential intracellular MuSK binding protein, while monovalent antibodies inhibited agrin-induced phosphorylation of MuSK, thus demonstrating distinct molecular mechanisms underlying the MuSK dysfunction induced by these two types of antibodies. Taken together, these findings suggest that complement activation is not necessary for the MG onset and that both divalent and monovalent antibodies may cause MG in vivo by inducing MuSK dysfunction.

Diurnal rhythms are altered in a mouse model of multiple sclerosis - Corrected Proof

Abigail C. Buenafe — 2012-01-03

Abstract: Our earlier studies described a disruption of heart rate and blood pressure diurnal rhythms in mice with experimental autoimmune encephalomyelitis (EAE). The present study investigates whether these observations could be extended to additional clock-regulated rhythms in mice with EAE. Analysis of clock gene expression in the liver of EAE mice demonstrated significant variability associated with Per2 rhythmic expression. Corticosterone and leptin hormone rhythms were also altered in EAE mice. The results presented here indicate that disturbances in clock-regulated rhythms are associated with EAE and present a suitable model for investigating the relationship between circadian disruption and autoimmune inflammatory disease.

S100B modulates growth factors and costimulatory molecules expression in cultured human astrocytes - Corrected Proof

2011-12-26

Abstract: S100B is a Ca2+-binding protein expressed in the nervous system. Increased levels of S100B in the extracellular space have been detected in several neurological disorders. We investigated the response of human astrocytes to micromolar chronic concentrations of this protein measuring the expression of some costimulatory molecules, such as CD137, CD137-L, CD40, CD40-L, the chemokine RANTES and two growth factors FGF-2 and TGF-β2. Our findings suggest that high levels of S100B in the brain parenchyma may modulate the activation status of astrocytes decreasing their neuroprotective role and modifying their interaction with microglia and other inflammatory cells. This effect may contribute to evolution of some neurological disorders.

A role for inflammatory mediators in the modulation of the neurotrophin receptor p75NTR on human muscle precursor cells - Corrected Proof

Emanuela Colombo, Stefania Romaggi, Marina Mora, Lucia Morandi, Cinthia Farina — 2011-12-26

Abstract: The neurotrophin receptor p75NTR is a marker for human differentiation-prone muscle satellite cells and regulates myogenesis. Here, we wondered whether inflammation could modify p75NTR expression on muscle precursor cells in vitro and in vivo. In vitro experiments on human primary skeletal myoblasts demonstrated that exposure to IFN-γ or IL-1α decreased transcript and protein levels of p75NTR. Furthermore, histological investigations showed a reduction in the pool of p75NTR expressing satellite cells in inflammatory myopathy biopsies. These data suggest a link between muscle inflammation and reduction of p75NTR expression on precursor cells.

Investigation of autoantibody profiles for cerebrospinal fluid biomarker discovery in patients with relapsing–remitting multiple sclerosis - Corrected Proof

2011-12-19

Abstract: Using the UNIarray® marker technology platform, cerebrospinal fluid immunoglobulin G reactivities of 15 controls and 17 RRMS patients against human recombinant proteins were investigated. Patient cerebrospinal fluids were oligoclonal band positive and reactivities were compared to that of sex- and age-matched controls. We hereby aimed at the characterization of autoreactivity in patients with RRMS. Differences in autoreactivities between control and RRMS samples were identified comprising autoantigens identified in this study only and previously reported autoantigens as well. A combination of the 10–15 most significant proteins may be investigated further as autoantigens for diagnostic purposes. Additional investigations may include minimizing the number of proteins used in such diagnostic tests.

Plasma sCD28, sCTLA-4 levels in neuromyelitis optica and multiple sclerosis during relapse - Corrected Proof

2011-12-19

Abstract: Background: Soluble forms of CD28 (sCD28) and CTLA-4 (sCTLA-4) were associated with many autoimmune diseases like Sjögren's syndrome, systemic lupus erythematosus, asthma, and autoimmune myasthenia gravis. However, sCD28 and sCTLA-4 in neuromyelitis optica (NMO) and multiple sclerosis (MS) patients were less studied.Objective: To measure the plasma sCD28, sCTLA-4 in NMO and MS patients, and investigate whether sCD28 and sCTLA-4 possible use as sensitive biomarkers for diseases activity.Methods: Plasma concentrations of sCD28, sCTLA-4 were measured by an enzyme-linked immunosorbent assay (ELISA) in NMO (n=22), MS (n=21) patients and controls (n=18).Results: The concentration of sCD28 levels were higher in the inflammatory demyelinating diseases cohort compared with the controls (NMO, p=0.034; MS, p=0.026) and the levels of sCD28 were slightly higher in NMO compared with MS. The sCTLA-4 levels were lower in the MS subgroup compared with the controls (p=0.032). Both sCD28 and sCTLA-4 did not show any correlation with EDSS score in NMO and MS patients.Conclusions: Our study revealed for the first time that the levels of increased plasma sCD28 and decreased plasma sCTLA-4 in NMO and MS patients, but had little correlation with clinical presentations.

Guillain–Barre syndrome: First description of a snake envenomation aetiology - Corrected Proof

2011-12-16

Abstract: Background: Guillain–Barre syndrome (GBS) is considered as an acute, immune-mediated polyradiculoneuropathy with different clinical phenotypes arising after viral or bacterial infections, vaccination or surgery. However, in 40% of GBS patients the aetiology remains unknown. In this manuscript, we report the occurrence of GBS in a patient bitten by a snake (Vipera aspis) for which a cross-reaction was shown between GM2 ganglioside and glycosidic epitopes of venom proteins.Methods: The venom of the snake implied in the patient's envenomation was collected. Its composition was characterised by ELISA and SELDI-TOF MS. Cross-reactivities between venom proteins and GM2 gangliosides were identified by Western blot after immunoabsorption of patient's serum with increasing amounts of purified GM2. Enzymatic deglycosylation of the venom was performed to determine the specificity of the patient's serum cross-reaction.Findings: We proved the absence of neurotoxicity of the viper venom. The patient's serum presented specific cross-reactions with several glycosylated venom proteins. After deglycolysation of these proteins, the patient's serum cross-reactivity was abolished. Furthermore, we compared the immune response to venom proteins of sera from two groups of patients. The first group showed IgM reactivity against GM2 ganglioside associated with GBS, and cross-reacted with venom proteins. The second group presented an IgM reactivity against CMV, without neurological disorders, and reacted with neither venom proteins nor gangliosides.Interpretation: Our study proved the auto-immunological aetiology of GBS in our patient based on molecular mimicry mechanisms between venom proteins and GM2 ganglioside.

Neutrophils in multiple sclerosis are characterized by a primed phenotype - Corrected Proof

2011-12-12

Abstract: Neutrophils are armed with proteases with indiscriminate histotoxic potential, and to minimize tissue injury, their activation involves priming with inflammatory mediators before cells are fully activated in a second step. Here, we show that neutrophils in multiple sclerosis patients are more numerous and exhibit a primed state based on reduced apoptosis, higher expression of TLR-2, fMLP receptor, IL-8 receptor and CD43, enhanced degranulation and oxidative burst as well as higher levels of neutrophil extracellular traps in serum. The chronic inflammatory environment in multiple sclerosis probably underlies this inappropriate neutrophil priming, which may result in enhanced neutrophil activation during infection.

Immune profiling of Alzheimer patients - Corrected Proof

2011-12-08

Abstract: Alzheimer's disease (AD) is characterized by extracellular senile plaques in the brain, containing amyloid-β peptide (Aβ). We identify immunological differences between AD patients and age-matched controls greater than those related to age itself. The biggest differences were in the CD4+ rather than the CD8+ T cell compartment resulting in lower proportions of naïve cells, more late-differentiated cells and higher percentages of activated CD4+CD25+ T cells without a Treg phenotype in AD patients. Changes to CD4+ cells might be the result of chronic stimulation by Aβ present in the blood. These findings have implications for diagnosis and understanding the aetiology of the disease.

Synapsin peptide fused to E. coli heat-labile toxin B subunit induces regulatory T cells and modulates cytokine balance in experimental autoimmune encephalomyelitis - Corrected Proof

M. Julia Scerbo, Mario J. Bibolini, German A. Roth, Clara G. Monferran — 2011-12-06

Abstract: We previously found that the preventive oral administration of a hybrid consisting of the C domain of synapsin and the B subunit of E. coli heat-labile enterotoxin (LTBSC) efficiently suppresses experimental autoimmune encephalomyelitis (EAE) development in rats. We investigated the effect of LTBSC on cytokine expression and on regulatory T (Treg) cells in rats with myelin induced EAE. LTBSC treatment increased the frequency of CD4+FoxP3+ Treg cells in lymph nodes prior to challenge and in the EAE acute stage. LTBSC also up-regulated the expression of anti-inflammatory Th2/Th3 cytokines and diminished myelin basic protein-specific Th1 and Th17 cell responses in lymph nodes. CD4+CD25+ Treg cells from LTBSC treated rats showed stronger suppressive properties than Treg cells from controls in vitro. Our observations indicate that LTBSC is a useful agent for modulating the autoimmune responses in EAE.

The role of the Epstein–Barr Virus receptor CD21 in Multiple Sclerosis - Corrected Proof

Nicole Toepfner, Sabine Cepok, Verena Grummel, Bernhard Hemmer — 2011-12-02

Abstract: Multiple Sclerosis (MS) is characterised by a chronic inflammation and demyelination of brain and spinal cord with a yet unknown aetiology. Based on multiple epidemiological and immunological studies, which suggest a role of Epstein–Barr virus (EBV) infection in the pathogenesis of MS, we investigated CD21 (CR2, complement receptor type 2), which serves as the EBV receptor. Serum concentrations of soluble CD21 receptor (sCD21) were determined in MS patients and controls. In accordance with findings in other autoimmune disorders decreased sCD21 levels were found in MS patients. On ß-IFN treatment serum sCD21 concentrations further decreased. To explore the role of the CD21 gene for MS susceptibility and the altered CD21 levels in MS patients we performed exon sequencing of the CD21 gene. While we identified new single nucleotide polymorphism (SNP) and confirmed previously reported SNPs, none of the SNPs was associated with MS. Our findings demonstrate that sCD21 expression is altered in MS patients similar to other autoimmune diseases although no evidence was found for a specific role of the CD21 gene in MS.

Proteomics technologies for biomarker discovery in multiple sclerosis - Corrected Proof

Vaibhav Singh, Rogier Q. Hintzen, Theo M. Luider, Marcel P. Stoop — 2011-12-02

Abstract: Multiple sclerosis is a disabling inflammatory and neurodegenerative disorder that predominantly affects young adults. There is a great need for biomarkers, which could elucidate pathology as well as provide prognosis of disease progression and therapy response in multiple sclerosis. Rapidly evolving, technology driven applications such as mass spectrometry based proteomics are currently being developed for this purpose. In this review, we will outline the current status of the field and detail a number of the bottlenecks as well as future prospects of this type of biomarker research.

Evidence of central inflammation in fibromyalgia — Increased cerebrospinal fluid interleukin-8 levels - Corrected Proof

Diana Kadetoff, Jon Lampa, Marie Westman, Magnus Andersson, Eva Kosek — 2011-11-30

Abstract: Activation of glia cells resulting in intrathecal elevation of cytokines and chemokines has been hypothesized in chronic pain syndromes such as fibromyalgia. To our knowledge, this is the first study assessing intrathecal concentrations of pro-inflammatory substances in fibromyalgia. We report elevated cerebrospinal fluid and serum concentrations of interleukin-8, but not interleukin-1beta, in FM patients. This profile is in accordance with FM symptoms being mediated by sympathetic activity rather than dependent on prostaglandin associated mechanisms and supports the hypothesis of glia cell activation in response to pain mechanisms.

Anti-glioma response of autologous T cells stimulated by autologous dendritic cells electrofused with CD133+ or CD133− glioma cells - Corrected Proof

2011-11-28

Abstract: Glioma, the most common tumor of the central nervous system (CNS), currently results in a high rate of morbidity and mortality. The expression of CD133, a stem-like cell marker expressed in the glioma cells, is believed to lead to tumorigenesis in the human brain. Thus, it is necessary to find a proper method to specifically kill the CD133+ glioma cells. Dendritic cell (DC)/tumor hybrids are proven to be able to induce an effective immune response, leading to killing of glioma cells in vitro. We isolated CD133+ cells from a population of primary glioma cells, and cultured autologous DCs and T cells at the same time. Next, we electrofused the DCs with the CD133+ glioma cells and with CD133− ones, in order to explore a new strategy for glioma therapy. We then exposed the T cells to five separate groups of cells: DC/CD133+ hybrids, DC/CD133− hybrids, DCs alone, unsorted glioma cells alone and mixed DCs-glioma cells. A cytotoxicity assay showed that T cells stimulated by either type of hybrid were able to kill cultured autologous glioma cells significantly more effectively than those stimulated by the other three cell types (P<0.05). The amounts of IFN-γ secreted by T cells stimulated by the two types of fused cells were obviously increased compared to those stimulated by the other three cell types (P<0.05). However, no significant differences were noted between the effects of the two hybrids, neither in the cytotoxicity assay nor in the IFN-γ release assay (P>0.05). Therefore, both DC/CD133+ and DC/CD133− hybrids can cause significant T cell immune responses in vitro. There were no significant differences between the immune responses caused by the two types of hybrids.

Publication guidelines for refereeing and reporting on animal use in experimental autoimmune encephalomyelitis - Corrected Proof

David Baker, Sandra Amor — 2011-11-28

Abstract: Experimental Autoimmune encephalomyelitis (EAE) is one of the major pre-clinical models of multiple sclerosis. Use of this model has led to some important observations that has benefited human health, but it must also be said that there are many poor-quality studies using EAE that add little to knowledge or understanding. The lack of quality of clinical trials in humans led to the introduction of publication guidelines that aimed to improve the quality and transparency of studies. Therefore, with a view of doing the same, the ARRIVE (Animals in research: Reporting in vivo experiments) guidelines were developed as a checklist of information required for publication of animal studies. Based on the ARRIVE guidelines, we provide guidance on acceptable data handling and indicate the minimal acceptable standard that should be adhered to when reporting and importantly refereeing all publications relating to EAE. This is designed to constructively improve the quality of published work and reduce the futile use of animals in research.

Substance P–Neurokinin-1 receptor interaction upregulates monocyte tissue factor - Corrected Proof

Mohammad M. Khan, Steven D. Douglas, Tami D. Benton — 2011-11-24

Abstract: Monocytes play an important role in hemostasis. In this study, the prothrombotic effects of the neuropeptide substance P (SP) on human monocytes through neurokinin-1 receptor (NK1-R) were characterized. SP upregulated monocyte tissue factor (TF), the major coagulation cascade stimulator, in a concentration and time dependent manner. Specific inhibition of NK1-R completely blocked TF expression. Monocytes stimulated by SP released cytokines and chemokines. When monocytes were stimulated with cytokines or chemokines, TF was expressed by the cytokines (GM-CSF, IFN-γ and TNF-α). Cytokines may play a major role in the mechanism of SP induced monocyte TF expression. NK1-R antagonists (NK1-RA) may have a role in developing novel therapeutic approaches to patients vulnerable to vaso-occlusive disorders.

IFN-β induces the proliferation of CD4+CD25+Foxp3+ regulatory T cells through upregulation of GITRL on dendritic cells in the treatment of multiple sclerosis - Corrected Proof

Meiyue Chen, Guangjie Chen, Shaohua Deng, Xin Liu, George J. Hutton, Jian Hong — 2011-11-23

Abstract: IFN-β is a major disease-modifying agent used for the treatment of multiple sclerosis (MS). Its mechanisms are complex and it has broad immunomodulatory effects on many types of immune cells. It was observed clinically that the quantity of CD4+CD25+Foxp3+ regulatory T cells increases in some MS patients treated with IFN-β. In this study, we show that IFNAR engagement by IFN-β expands naturally occurring CD4+CD25+Foxp3+ regulatory T cell population through the modulation of dendritic cells (DCs). IFN-β has no effect on the conversion of CD4+CD25− T cells to adaptive Treg cells. The IFN-β-induced upregulation of GITRL on DC and downregulation of CTLA-4 on Treg cell work together to facilitate the proliferation of anergic Treg cells. In MS patients treated with Avonex or Rebif (IFN-β), it was found that GITRL expression is markedly upregulated on peripheral CD14+ cells. Our findings help the better understanding of the complex effects of IFN-β in the treatment of MS.

miRNA profiling for biomarker discovery in multiple sclerosis: From microarray to deep sequencing - Corrected Proof

2011-11-11

Abstract: MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at the post-transcriptional level. miRNAs are highly expressed in cells of the immune and nervous system, attesting to their importance in Neuroimmunology. Besides their involvement in modulation of physiological and pathological processes, miRNAs hold high promise as disease biomarkers, therapeutic agents and/or drug targets. Several studies have recently explored the involvement of miRNAs in Multiple Sclerosis (MS) using a variety of miRNA profiling techniques. In this review, we discuss basic miRNA biology and nomenclature, the techniques available for miRNA profiling research and recent miRNA profiling studies in Multiple Sclerosis.

Chemokines and chemokine receptors in the nervous system - Corrected Proof

2007-05-11