Response of Lymphocytes from Multiple Sclerosis Patients to Murine Brain Extract

Forty-seven patients with multiple sclerosis (MS), 21 subjects with neurological diseases other than MS (OND), 7 with miscellaneous diseases (MD) and 21 healthy individuals (HI) were examined by the standard procedure of blast transformation for the in vitro response to mouse brain (MB), mouse kidney (MK) and mouse lung (ML) tissue extracts. Increase cellular response to MB was limited to the MS and OND groups, whereas high and low values of stimulation index to MK and ML were similarly observed in all groups of individuals examined. No correlation was found between the clinical activity of the MS patients and the in vitro response to MB. The possibility that low sensitivity of the technique and complexity of the antigenic composition of MS are determining factors in the results obtained in these studies is discussed.     

Stress and Course of Disease in Multiple Sclerosis

Abstract

In this prospective study, 96 healthy controls and 101 multiple sclerosis patients were followed up for as many as 6 years, and self-reported stressful events and health status were assessed. The authors evaluated (a) whether patients reported more stressful life events than healthy controls and (b) the bidirectional relationship between stress and functional deterioration among patients. Healthy controls reported more life events than patients, Odds ratio (OR) = 1.13, p < .0001; and this relationship was attributable to healthy controls' reporting more neutral/positive events than patients. A bidirectional relationship was confirmed between stress and illness: there was an increased risk of disease progression when rate of reported stressful events was higher, OR = 1.13, p < .0003, and an increased risk of reported stressful events when rate of disease progression was higher, OR = 2.13, p < .0001. There were no differences in reported stress by level of baseline disability. The authors concluded that multiple sclerosis patients demonstrate a vicious cycle between stress and disease progression.     

Polyomavirus Models of Brain Infection and the Pathogenesis of Multiple Sclerosis

Multiple sclerosis (MS) is generally considered to be an autoimmune disorder with myelin as the target and with several unidentified viruses playing ancillary roles, possibly through molecular mimicry. Although this paradigm has led to important progress on potential mechanisms of myelin loss, neither a target antigen in myelin nor a triggering mechanism has yet been identified, leaving the etiology of MS still unknown. Animal models of viral demyelination and studies showing that JC virus (JCV), the polyomavirus which causes progressive multifocal leukoencephalopathy (PML), may be latent in some normal human brains suggest another possibility. A host immune response targeting proteins expressed at low levels from viral DNA latent in the central nervous system (CNS) might underlie a focal demyelinating disease such as MS. A shift from autoimmunity to a latent-virus model is not a trivial substitution of target antigens. This shift would expand the search for a definitive laboratory test for MS and could lead to improved therapeutic and preventive approaches.     

Focal Slow and Beta Brain Activity in Patients with Multiple Sclerosis Revealed by Magnetoencephalography

In multiple sclerosis (MS) inflammatory infiltrations cause white matter lesions. Magnetoencephalography (MEG) offers the opportunity to localize abnormal electric activity of neurons with a high spatio-temporal resolution. In this study, we investigated patients with MS in order to find if abnormal cortical activity is associated with (subcortical) MS lesions using simultaneous bilateral recording of biomagnetic activity. Eight patients suffering from definite laboratory-supported MS with mainly somatosensory deficits and multiple bihemispheric plaques revealed by MRI were included in the study. To obtain normative data, 8 healthy volunteers were investigated following the same measuring protocol. Spontaneous magnetic brain activity was recorded using a 2×37-channel biomagnetic system (BTI, USA). Offline analysis included digital filtering (to separately investigate slow and beta wave activity), a Principle Component Analysis and the Dipole Density Plot. Localization results were inserted into MR images using our contour fit procedure. The dipole distribution in the brain was quantified and compared between the groups by statistical analysis. In all MS patients, the maximum of focal abnormal activity was localized in cortical areas adjacent to the fiber lesions. In the healthy subjects, no focal abnormal brain activity could be found. However, the standardized maximum concentrations of dipoles were significantly higher in the MS patients than in the healthy control group both in the slow and in the beta wave analysis. These results let assume that subcortical lesions can occur together with abnormal cortical neuronal activity. The results are discussed in respect to their impact on the interpretation of the analysis of spontaneous magnetic brain activity.     

Immunoglobulin Synthesis In Vitro by Cerebrospinal Fluid Cells in Patients with Multiple Sclerosis

Incubated cerebrospinal fluid (CSF) cells from patients with multiple sclerosis synthesized IgG and IgA in vitro. The synthesized IgG had an oligoclonal distribution and showed the same elcctrophoretic pattern as the IgG of the CSF. The amount synthesized was greater during exacerbations than during remissions. Blood lymphocytes from the same patients synthesized an IgG in vitro that showed a completely different electrophoretic pattern The amount IgG synthesized by the blood lymphocytes was less than the amount synthesized by the CSF cells. The results demonstrate that at least part of the oligoclonal IgG of the CSF of patients with multiple sclerosis is synthesized intrathecally and suggest that the CSF cells are antigenically stimulated in vivo.     

Correlation Between Interferon Production and Clinical Disease Activity in Patients with Multiple Sclerosis

We determined the interferon (IFN) serum levels and in vitro activated IFN production in eight patients with relapsing/remitting multiple sclerosis (MS), using a whole-blood test system and the mitogen concanavalin A and the viral antigen Newcastle disease virus for induction of the IFN production. During the overall study period of 12 months we observed, in relation to clinical disease progression, a biphasic increase in the individual IFN and IFN production. While mitogen-induced IFN synthesis showed a significant augmentation prior to the onset of a new relapse (P < 0.05), virus-induced IFN production showed a temporal delayed increase which was related to clinical remission (P < 0.01). The observed fluctuations in the individual production of both IFN subtypes were not reflected in the sera of the patients. Although the reason for the temporal different imbalance in the production of both IFN subtypes remains unknown, the observed association between increased IFN production and clinical remission emphasizes a possible role for type 1 IFNs in the resolution of the MS relapse.     

Oligodendrocyte Lineage Cells in Chronic Demyelination of Multiple Sclerosis Optic Nerve

Reports that chronically demyelinated multiple sclerosis brain and spinal cord lesions contained immature oligodendrocyte lineage cells have generated major interest aimed at the potential for promotion of endogenous repair. Despite the prominence of the optic nerve as a lesion site and its importance in clinical disease assessment, no detailed studies of multiple sclerosis‐affected optic nerve exist. This study aims to provide insight into the cellular pathology of chronic demyelination in multiple sclerosis through direct morphological and immunohistochemical analysis of optic nerve in conjunction with observations from an experimental cat optic nerve model of successful remyelination. Myelin staining was followed by immunohistochemistry to differentially label neuroglia. Digitally immortalized sections were then analyzed to generate quantification data and antigenic phenotypes including maturational stages within the oligodendrocyte lineage. It was found that some chronically demyelinated multiple sclerosis optic nerve lesions contained oligodendroglial cells and that heterogeneity existed in the presence of myelin sheaths, oligodendrocyte maturational stages and extent of axonal investment. The findings advance our understanding of oligodendrocyte activity in chronically demyelinated human optic nerve and may have implications for studies aimed at enhancement of endogenous repair in multiple sclerosis.     

Increased Spontaneous Ex Vivo Apoptosis and Subset Alterations in Peripheral Blood T Cells from Patients with Multiple Sclerosis

  In order to characterize the immunophenotype and the lymphocyte apoptosis in multiple sclerosis (MS) patients, the peripheral blood lymphocytes of 46 MS patients and 12 healthy volunteers were studied by flow cytometry. Immunophenotypic alterations included significant increases in T CD4+ lymphocytes and reductions in the percentages of CD3+ and CD8+ T cells. After 24 h of culture spontaneous apoptosis was increased in almost T lymphocyte subsets from MS patients and it was significantly higher in those patients who had suffered more than two relapses in the two previous years. The incidence of spontaneous apoptosis was not dependent on FasL-Fas interactions and correlated with the percentages of cells positive for active caspases but not with percentages of Fas+ cells. The increased susceptibility to apoptosis of peripheral blood T lymphocytes from MS patients is difficult to reconcile with the previously proposed role of a defective lymphocyte apoptosis in the pathophysiology of MS.Alfredo Prieto and David Díaz contributed equally to this work and are joint first authors.The GENIO-II Group is composed by Pablo Barreiro from Hospital La Paz, Madrid; Clara de Andrés and Ma Luisa Martínez from Hospital Gregorio Marañón, Madrid; Bonaventura Casanova from Hospital La Fe, Valencia; Juan Antonio García-Merino and Rosario Blanco from Hospital Puerta de Hierro, Madrid; Jesús Martín from Hospital Miguel Servet, Zaragoza; Francisco Coret from Hospital Clínico, Valencia; José Carlos Alvarez-Cermeño and Dr José Francisco Plaza from Hospital Ramón y Cajal, Madrid; Eugenia Vilar, Blanca Felgueroso and Julián Benito from Hospital de Móstoles; Asunción Morales Otal and M^a Cruz Gutiérrez del Olmo from Hospital 12 de Octubre, Madrid.     

Leptin Enhances the Release of Cytokines by Peripheral Blood Mononuclear Cells from Relapsing Multiple Sclerosis Patients

Leptin, a hormone synthesized mainly by adipocytes, can modulate the immune response and seems to be involved in the induction of experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis (MS). However, the possible role of leptin in MS pathogenesis has not yet been elucidated. In this study we investigated the effect of leptin on cytokine production by peripheral blood mononuclear cells (PBMCs) of MS patients (either in the acute or in the stable phase of the disease) and healthy controls. We also analyzed leptin effects on cytokine production by monocytes in relapsing MS patients. Our data showed that leptin induced tumor necrosis factor-alpha, interleukin-6, and interleukin-10 production by PBMCs of patients in an acute phase of disease but not in patients in a stable phase or in healthy controls. Moreover, we found no effect of leptin in monocytes from relapsing MS patients. Therefore we conclude that leptin may modulate the MS inflammatory process during relapses.     

Axon Reactive B Cells Clonally Expanded in the Cerebrospinal Fluid of Patients with Multiple Sclerosis

Demyelination and axonal loss have been described as the histological hallmarks of inflammatory lesions of multiple sclerosis (MS) and are the pathological correlates of persistent disability. However, the immune mechanisms underlying axonal damage in MS remain unknown. Here, we report the use of single chain-variable domain fragments (scFv) from clonally expanded cerebrospinal fluid (CSF) B cells to show the role of an anti-axon immune response in the central nervous system (CNS) in MS. The cellular and subcellular distribution of the antigen(s) recognized by these CSF-derived clonal scFv antibodies (CSFC-scFv Abs) was studied by immunochemical staining of brain tissues obtained at autopsy from patients with MS. Immunochemistry showed specific binding of CSFC-scFv Abs to axons in acute MS lesions. The stained axons showed three major types of axonal pathological changes: 1) linear axons, axonal ovoid formation, and axonal transection were seen in the myelinated white matter adjacent to the lesion; 2) accumulation of axonal ovoid formations and Wallerian degeneration were seen at the border between demyelinated lesions and the adjacent white matter; and 3) Wallerian degeneration occurred at the center and edge of acute demyelinated lesions. These findings suggest a B cell axonal specific immune response in the CNS in MS.     

IgG1 Subclass Restriction of Oligoclonal IgG from Cerebrospinal Fluids and Brain Extracts in Patients with Multiple Sclerosis and Subacute Encephalitides

Oligoclonal-IgG-containing cerebrospinal fluids (CSF) from patients with multiple sclerosis and subacute encephalitides were studied for IgG subclass distribution by immunoelectrophoretic and hemagglutination inhibition methods. The immunoelectrophoretic results indicated the presence of electrophoretically restricted IgG1 proteins in a number of CSF, compatible with an association between this IgG subclass and oligoclonal IgG proteins. The combined results indicated a greater dominance of IgG1 over other IgG subclass proteins in CSF than in matching sera. Similar results were obtained in experiments with brain extracts from patients with subacute sclerosing panencephalitis. The results differed from those obtained with normal CSF, where the distribution of IgG subclasses resembled that of the matching sera. It is concluded that the oligoclonal IgG of the CSF and brain in the patients studied belong mainly to the IgG1 subclass.     

Protective and Detrimental Roles for Regulatory T Cells in a Viral Model for Multiple Sclerosis

Multiple sclerosis (MS) has been proposed to be an immune‐mediated disease in the central nervous system (CNS) that can be triggered by virus infections. In Theiler's murine encephalomyelitis virus (TMEV) infection, during the first week (acute stage), mice develop polioencephalomyelitis. After 3 weeks (chronic stage), mice develop immune‐mediated demyelination with virus persistence, which has been used as a viral model for MS. Regulatory T cells (Tregs) can suppress inflammation, and have been suggested to be protective in immune‐mediated diseases, including MS. However, in virus‐induced inflammatory demyelination, although Tregs can suppress inflammation, preventing immune‐mediated pathology, Tregs may also suppress antiviral immune responses, leading to more active viral replication and/or persistence. To determine the role and potential translational usage of Tregs in MS, we treated TMEV‐infected mice with ex vivo generated induced Tregs (iTregs) on day 0 (early) or during the chronic stage (therapeutic). Early treatment worsened clinical signs during acute disease. The exacerbation of acute disease was associated with increased virus titers and decreased immune cell recruitment in the CNS. Therapeutic iTreg treatment reduced inflammatory demyelination during chronic disease. Immunologically, iTreg treatment increased interleukin‐10 production from B cells, CD4⁺ T cells and dendritic cells, which may contribute to the decreased CNS inflammation.     

Access to health services in Ireland for people with Multiple Sclerosis and Motor Neurone Disease

We conducted a telephone questionnaire to determine the utilisation of hospital and community based services by patients with Motor Neurone Disease and Multiple Sclerosis in Ireland. 94 MND and 188 MS patients participated in the study. MND patients were more likely to have free medical care than MS patients, despite legislation favouring the converse. Severely disabled MND patients were more successful at accessing free community-based services than were severely disabled MS patients. Private medical insurance conferred no advantage when obtaining services or purchasing equipment. Many patients were unaware of the specific roles of the various clinical professionals. There are significant deficiencies in patients' ability to access multidisciplinary services. Voluntary organisations often bridge the gap in service provision. An investment in services for people with chronic neurological disability is urgently required.     

Levels of Interleukin-15-Expressing Blood Mononuclear Cells Are Elevated in Multiple Sclerosis

Interleukin-15 (IL-15) is a novel IL-2-like cytokine expressed by cells of the monocyte/macrophage and epithelial lineages. Cytokines might be involved in the pathogenesis of multiple sclerosis (MS). Using immunocytochemistry, we analysed spontaneous expression of IL-15 by peripheral blood (PB) and cerebrospinal fluid (CSF) mononuclear cells (MNC) from patients with MS, other neurological diseases (OND) and healthy controls. IL-15- positive peripheral blood mononuclear cells (PBMNC) were elevated in patients with MS compared to healthy controls (P < 0.05). The elevation of IL-15- positive PBMNC was restricted to patients with chronic progressive MS and not observed in patients studied during the relapsing-remitting phase of MS. The numbers of IL-15- expressing PBMNC correlated with the duration and disability of MS (r = 0.45, P < 0.001, and r = 0.39, P < 0.01, respectively). IL-15 was undetectable in CSF MNC, and ELISA showed low CSF levels of IL-15 in occasional patients with MS and OND. IL-15 is a potent growth factor for gammadelta T cells, but there was no correlation between IL-15 expression by PBMNC and percentage of gammadelta T cells in blood from the MS patients. Together, these data demonstrate that IL-15 expression by PBMNC is upregulated in the chronic stage of MS.