Animal Model Of MS Research Articles

Expression of GM-CSF in T Cells Is Increased in Multiple Sclerosis and Suppressed by IFN-β Therapy.

Multiple sclerosis (MS) is an autoimmune disease of the CNS. Studies in animal models of MS have shown that GM-CSF produced by T cells is necessary for the development of autoimmune CNS inflammation. This suggests that GM-CSF may have a pathogenic role in MS as well, and a clinical trial testing its blockade is ongoing. However, there have been few reports on GM-CSF production by T cells in MS. The objective of this study was to characterize GM-CSF production by T cells of MS patients and to determine the effect of IFN-β therapy on its production. GM-CSF production by peripheral blood (PB) T cells and the effects of IFN-β were characterized in samples of untreated and IFN-β-treated MS patients versus healthy subjects. GM-CSF production by T cells in MS brain lesions was analyzed by immunofluorescence. Untreated MS patients had significantly greater numbers of GM-CSF(+)CD4(+) and CD8(+) T cells in PB compared with healthy controls and IFN-β-treated MS patients. IFN-β significantly suppressed GM-CSF production by T cells in vitro. A number of CD4(+) and CD8(+) T cells in MS brain lesions expressed GM-CSF. Elevated GM-CSF production by PB T cells in MS is indicative of aberrant hyperactivation of the immune system. Given its essential role in animal models, abundant GM-CSF production at the sites of CNS inflammation suggests that GM-CSF contributes to MS pathogenesis. Our findings also reveal a potential mechanism of IFN-β therapy, namely suppression of GM-CSF production.

Expression of GM-CSF in T cells is increased in multiple sclerosis and is suppressed by IFN-β therapy (BA11P.136)

Abstract Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS). Studies in animal models of MS have shown that granulocyte-macrophage colony-stimulating factor (GM-CSF) produced by T cells is necessary for development of autoimmune CNS inflammation. This suggests that GM-CSF may have a pathogenic role in MS as well. The objective of this study was to characterize GM-CSF production by T cells of MS patients, and to determine the effect of interferon-beta (IFN-β) therapy on its production. GM-CSF production by peripheral blood (PB) T cells and the effects of IFN-β were characterized in samples of untreated and IFN-β-treated MS patients vs. healthy subjects. GM-CSF production by T cells in MS brain lesions was analyzed by immunofluorescence. Untreated MS patients had significantly greater numbers of GM-CSF+ CD4+ and CD8+ T cells in PB compared to healthy controls and IFN-β-treated MS patients. IFN-β strongly suppressed GM-CSF production by T cells in vitro. A number of CD4+ and CD8+ T cells in MS brain lesions expressed GM-CSF. Elevated GM-CSF production by PB T cells in MS is indicative of aberrant hyperactivity of the immune system. Given its essential role in animal models, abundant GM-CSF production at the sites of CNS inflammation suggests that GM-CSF contributes to MS pathogenesis. Our findings also reveal a potential mechanism of IFN-β therapy, namely suppression of GM-CSF production.

MHC-mismatched mixed chimerism augments thymic regulatory T cell production and prevents relapse of EAE (THER7P.960)

Abstract Multiple sclerosis (MS) is an autoimmune inflammatory disease in central nervous system with demyelination, axon damage, and paralysis. There is no yet curative therapy. Induction of MHC-mismatched mixed chimerism is proposed to be a potential cure for autoimmune diseases, but this has not yet been tested in animal models of MS, experimental autoimmune encephalomyelitis (EAE). Here, we report that MHC-mismatched mixed chimerism with C57BL/6 (H-2b) donor in SJL/J (H-2s) EAE recipients eliminates symptoms and prevents relapse. This cure is demonstrated by not only disappearance of clinical signs but also reversal of autoimmunity and elimination of infiltration of T, B and macrophage cells in the spinal cord, as judged by HE, immunofluorescent staining and flow cytometry analysis. We also see regeneration of myelin as judged by fast-blue staining and prevention of axon damage as judged by electronic microcopy. The reversal of autoimmunity is associated with a marked reduction of autoreactivity of CD4+ T cells and significant increase of Foxp3+ regulatory T percentage among host-type CD4+ T cells in the spleen and lymph nodes. We have also found a marked increase of Foxp3+ regulatory T percentage among host-type CD4+ and CD4+CD8+ thymocytes in the thymus. Thymectomy leads to loss of prevention of EAE relapse by the mixed chimerism. These results indicate that induction of MHC-mismatched mixed chimerism can augment thymic production of Foxp3+ regulatory T cells and cure EAE.

GM-CSF production by CD4+ T cells in MS patients: regulation by regulatory T cells and vitamin D.

Data from animal models of MS suggest that GM-CSF(+)CD4(+)T cells are pathogenic cells. Therefore, GM-CSF production by CD4(+)T cells of MS patients and their susceptibility to regulatory mechanisms were investigated. Intracellular flowcytometry was performed to determine the GM-CSF(+)CD4(+)T cell fraction in PBMC and CSF of MS patients and controls. The effect of regulatory T cells (Tregs) on GM-CSF production by CD4(+)T cells was studied in MS patients using a proliferation-suppression assay. Finally, GM-CSF(+)CD4(+)T cell fraction and GM-CSF protein levels in supernatant were assessed in anti-CD3-stimulated CD4(+)T cell cultures derived from healthy controls and MS patients, in the presence or absence of the active vitamin D metabolite calcitriol. The GM-CSF(+)CD4(+)T cell fraction in the peripheral blood did not differ between controls and MS patients. This T cell population could also be detected in the CSF of both subjects with MS as well as subjects with another diagnosis. In the CSF, it comprised a significant fraction of the T cell population. Upon in vitro stimulation of PBMC with anti-CD3 antibody, no differences were observed in GM-CSF(+)CD4(+)T cell frequencies. GM-CSF secretion was susceptible to regulation by Treg and vitamin D. Suppression of GM-CSF secretion by vitamin D was reduced in MS patients. Our study showed no elevation in GM-CSF(+)CD4(+)T cell fractions in MS patients compared to controls. Furthermore, GM-CSF secretion was prone to regulation by Treg and vitamin D, the latter being less effective in MS patients.

Mechanism of multiple sclerosis based on the clinical trial results of molecular targeted therapy

AbstractMultiple sclerosis (MS) is a complex immune mediated disease of the central nervous system. It is characterized by inflammatory and neurodegenerative processes that result in neuroaxonal damage. Its etiology is still unknown, and its pathogenesis is only partly understood. There have been major advances in the treatment of MS in the past two decades, and a wide range of immunomodulagtory and immunosuppressive therapies have been used for the management of MS. More recently, there has been a growing interest in immunotherapeutic strategies with selective actions that target biological molecules involved in MS pathogenesis. Thus, better understanding of the immunopathogenesis of MS is believed to result in the development of more efficacious treatment. However, in contrast to the successful introduced therapies, such as natalizumab and alemtuzumab, there have been a remarkable number of therapeutic failures as well. Despite the convincing immunological concepts and promising results from animal models of MS, some drugs showed no clinical efficacy or even worsened the disease. Clinical trial results of molecular targeted therapy that shed light on the improving understanding of the immunopathogenesis of MS are discussed in the present review. These trials include monoclonal antibodies against leukocyte differentiation molecules (anti‐CD3 and anti‐CD4 antibodies), tumor necrosis factor‐α neutralization, targeting the interleukin (IL)‐12/IL‐23 pathways, immune cell‐depleting ant‐CD52 monoclonal antibody and targeting IL‐2 receptor signaling.

Elevated NKG2D ligand expression in experimental autoimmune encephalomyelitis

T cells remain unclear. Expressing myelin-reactive T cell receptor (TCR) is not sufficient to make a T cell encephalitogenic. In fact, the frequencies of myelin-reactive T cells are comparable between MS patients and healthy individuals, but the ones in MS patients have activated/memory phenotypes. In the animal model of MS, experimental autoimmune encephalomyelitis (EAE), myelin-specific T cells activated with antigen presenting cells (APCs) plus myelin peptide are encephalitogenic, whereas T cells activated with anti-CD3/CD28 antibodies are not. This suggests that APCs provide critical cytokines beyond T cell receptor activation and co-stimulation, contributing to encephalitogenicity. To identify which cytokines are critical for generating an encephalitogenic T cell, naive myelin-specific CD4 T cells were activated with anti-CD3/CD28 in the presence of various cytokines, which recapitulated the signals normally provided by APCs. The activated T cells were then adoptively transferred to wildtype recipients, and the degree of encephalitogenicity of T cells was examined by EAE. We showed that no single cytokine was sufficient, but the combinations of IL-6/IL-23 or IL-12/IL-23 generated highly encephalitogenic T cells. Previous studies have revealed an essential role of IL-23 in EAE pathogenesis. However, naive T cells differentiated with IL-23 alone could not transfer disease, due to limited expression of its receptor. IL-6 and IL-12 induced the initial expression of IL-23R on naive T cells, and IL-23 further enhanced the expression of its own receptor. IL-23R signaling induced downstream STAT3 phosphorylation in both IL-6/IL-23 and IL-12/ IL-23-generated T cells. Neutralizing IL-6 or IL-12 with antibodies significantly reduced the severity of EAE, which was induced by APCgenerated T cells, indicating the crucial roles of IL-6 and IL-12 during the initial T cell activation. We have identified that IL-6/IL-23 or IL-12/ IL-23 combination in addition to TCR activation and co-stimulation were the minimal signals necessary to generate encephalitogenic T cells. This provides potential targets that can be manipulated therapeutically, resulting in innovative treatments for MS.

Modulation of the kallikrein/kinin system by the angiotensin-converting enzyme inhibitor alleviates experimental autoimmune encephalomyelitis.

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS). Bradykinin is the end-product of the kallikrein/kinin system, which has been recognized as an endogenous target for combating CNS inflammation. Angiotensin-converting enzyme (ACE) inhibitors influence the kallikrein/kinin system and reportedly have immunomodulatory characteristics. The objectives of this study were to determine whether bradykinin is involved in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), an animal model of MS, and whether bradykinin control by the ACE inhibitor could be a therapeutic target in MS. The ACE inhibitor enalapril (1·0 or 0·2 mg/kg/day) was administered orally to EAE mice and the serum levels of bradykinin and cytokines in EAE mice were analysed. As a result, the administration of enalapril increased serum bradykinin levels, decreased the clinical and pathological severity of EAE and attenuated interleukin-17-positive cell invasion into the CNS. Additionally, bradykinin receptor antagonist administration reduced the favourable effects of enalapril. Our results suggest that bradykinin is involved in the pathomechanism underlying CNS inflammation in EAE, possibly through inhibiting cell migration into CNS. Control of the kallikrein/kinin system using ACE inhibitors could be a potential therapeutic strategy in MS.

Melatonin and metabolic regulation: a review.

Human life expectancy has increased over the past 50 years due to scientific and medical advances and higher food availability. However, overweight and obesity affect more than 50% of adults and 15% of infants and adolescents. There has also been a marked increase in the prevalence of metabolic syndrome in recent decades, which has been associated with a reduction in nocturnal pineal production of melatonin with aging and an increased risk of coronary diseases, type 2 diabetes mellitus (T2DM) and death. Melatonin is currently under intensive investigation in experimental animal models of diabetes, obesity and MS at pharmacological doses (between 5 and 20 mg kg(-1) body weight), demonstrating its capacity to ameliorate the total metabolic profile and its potential as an alternative to conventional drug therapies for the disorders associated with the MS, i.e. elevated systolic blood pressure, and impairment of glucose homeostasis, plasma lipid profile, inflammation, oxidative stress, and increased body weight. An especially significant finding is the induction by melatonin of white adipose tissue browning, which may be related to its effects against oxidative stress, uncoupling the mitochondrial bioenergetic process by enhancing the expression of uncoupled-protein-1 (UCP-1), which has been related to body weight reduction in experimental animals. Further research is required to improve knowledge of this mechanism. Clinical studies are needed with the administration of pharmacological melatonin doses, because the dose has ranged between 0.050 and 0.16 mg kg(-1) bw in most studies to date. Melatonin is a natural phytochemical, and it is also important to test its beneficial metabolic effects when consumed in functional foods.

Innate immune responses play a role in heterogeneity of EAE. (INM8P.445)

Abstract Multiple sclerosis (MS) is a heterogeneous disease, which makes treatment difficult. For example, responsiveness to IFNβ treatment among MS patients varies to a great extent. Using EAE as an animal model of MS, we found that IFNβ treatment is not effective when EAE develops in the absence of the NLRP3 inflammasome with alternative EAE induction methods. Importantly, innate immune responses triggered by alternative EAE induction methods played a role to generate different subtype of EAE. This led us to hypothesize EAE have subtypes (although they are not mutually exclusive), at least based on the dependency of the disease pathology on the NLRP3 inflammasome and on the response to IFNβ. In NLRP3 inflammasome-dependent IFNβ-responsive EAE (termed type-A EAE), the NLRP3 inflammasome plays a key role as a pathogenic factor of EAE by enhancing immune cell migration and the resulting immune cell recruitment in the CNS. On the other hand, the NLRP3 inflammasome is not essential for EAE induction, and IFNβ treatment is not effective in NLRP3 inflammasome-independent EAE (termed Type-B EAE). Type-A and Type-B EAE show clear difference in their phenotypes and pathological mechanisms, mediated by differential activation of innate immune responses. M.I. performed all the experiments and M.L.S. is the corresponding author of this study.

Matrine suppresses glutamate excitotoxicity in experimental autoimmune encephalomyelitis (HUM1P.323)

Abstract Multiple sclerosis (MS) is T cell-mediated inflammatory diseases characterized by lymphocyte infiltration and demyelination. Matrine (MAT) is a quinolizidine alkaloid derived from the herb Radix Sophorae Flave and has long been used in the treatment of hepatitis B in clinical without obvious side effects. Previous reports have revealed that MAT restrained the central inflammation after experimental autoimmune encephalomyelitis (EAE), the most common animal model of MS due to similar symptoms and mechanisms; however whether MAT is effective to inhibit glutamate excitotoxicity is still unclear. In this study, we provide evidence that MAT attenuated the EAE disease severity, accompanied by down-regulated glutamate level, increased GABA level, as well as promoted the expressions of two dependent glutamate transporters (GLT-1 and GLAST). In addition, MAT treatment significantly decreased the content of the NMDA- and AMPA- glutamate receptor in EAE rats. Together, these data indicate that MAT treatment inhibited glutamate excitotoxicity after EAE, and suggest that MAT may be a novel choice in MS therapy.

Myelin oligodendrocyte glycoprotein (MOG35-55) induced experimental autoimmune encephalomyelitis (EAE) in C57BL/6 mice.

Multiple sclerosis is a chronic neuroinflammatory demyelinating disorder of the central nervous system with a strong neurodegenerative component. While the exact etiology of the disease is yet unclear, autoreactive T lymphocytes are thought to play a central role in its pathophysiology. MS therapy is only partially effective so far and research efforts continue to expand our knowledge on the pathophysiology of the disease and to develop novel treatment strategies. Experimental autoimmune encephalomyelitis (EAE) is the most common animal model for MS sharing many clinical and pathophysiological features. There is a broad diversity of EAE models which reflect different clinical, immunological and histological aspects of human MS. Actively-induced EAE in mice is the easiest inducible model with robust and replicable results. It is especially suited for investigating the effects of drugs or of particular genes by using transgenic mice challenged by autoimmune neuroinflammation. Therefore, mice are immunized with CNS homogenates or peptides of myelin proteins. Due to the low immunogenic potential of these peptides, strong adjuvants are used. EAE susceptibility and phenotype depends on the chosen antigen and rodent strain. C57BL/6 mice are the commonly used strain for transgenic mouse construction and respond among others to myelin oligodendrocyte glycoprotein (MOG). The immunogenic epitope MOG35-55 is suspended in complete Freund's adjuvant (CFA) prior to immunization and pertussis toxin is applied on the day of immunization and two days later. Mice develop a "classic" self-limited monophasic EAE with ascending flaccid paralysis within 9-14 days after immunization. Mice are evaluated daily using a clinical scoring system for 25-50 days. Special considerations for care taking of animals with EAE as well as potential applications and limitations of this model are discussed.

Sex differences in a mouse model of multiple sclerosis: neuropathic pain behavior in females but not males and protection from neurological deficits during proestrus

BackgroundMultiple sclerosis (MS), a demyelinating disease of the central nervous system, is one of the most prevalent neurological disorders in the industrialized world. This disease afflicts more than two million people worldwide, over two thirds of which are women. MS is typically diagnosed between the ages of 20–40 and can produce debilitating neurological impairments including muscle spasticity, muscle paralysis, and chronic pain. Despite the large sex disparity in MS prevalence, clinical and basic research investigations of how sex and estrous cycle impact development, duration, and severity of neurological impairments and pain symptoms are limited. To help address these questions, we evaluated behavioral signs of sensory and motor functions in one of the most widely characterized animal models of MS, the experimental autoimmune encephalomyelitis (EAE) model.MethodsC57BL/6 male and female mice received flank injection of complete Freund’s adjuvant (CFA) or CFA plus myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) to induce EAE. Experiment 1 evaluated sex differences of EAE-induced neurological motor deficits and neuropathic pain-like behavior over 3 weeks, while experiment 2 evaluated the effect of estrous phase in female mice on the same behavioral measures for 3 months. EAE-induced neurological motor deficits including gait analysis and forelimb grip strength were assessed. Neuropathic pain-like behaviors evaluated included sensitivity to mechanical, cold, and heat stimulations. Estrous cycle was determined daily via vaginal lavage.ResultsMOG35-55-induced EAE produced neurological impairments (i.e., motor dysfunction) including mild paralysis and decreases in grip strength in both females and males. MOG35-55 produced behavioral signs of neuropathic pain—mechanical and cold hypersensitivity—in females, but not males. MOG35-55 did not change cutaneous heat sensitivity in either sex. Administration of CFA or CFA + MOG35-55 prolonged the time spent in diestrus for 2 weeks, after which normal cycling returned. MOG35-55 produced fewer neurological motor deficits when mice were in proestrus relative to non-proestrus phases.ConclusionsWe conclude that female mice are superior to males for the study of neuropathic pain-like behaviors associated with MOG35-55-induced EAE. Further, proestrus may be protective against EAE-induced neurological deficits, thus necessitating further investigation into the impact that estrous cycle exerts on MS symptoms.

Comparison of human adult stem cells from adipose tissue and bone marrow in the treatment of experimental autoimmune encephalomyelitis

IntroductionWhile administration of ex vivo culture-expanded stem cells has been used to study immunosuppressive mechanisms in multiple models of autoimmune diseases, less is known about the uncultured, nonexpanded stromal vascular fraction (SVF)-based therapy. The SVF is composed of a heterogeneous population of cells and has been used clinically to treat acute and chronic diseases, alleviating symptoms in a range of tissues and organs.MethodsIn this study, the ability of human SVF cells was compared with culture-expanded adipose stem cells (ASCs) and bone-derived marrow stromal cells (BMSCs) as a treatment of myelin oligodendrocyte glycoprotein (35–55)-induced experimental autoimmune encephalitis in C57Bl/6J mice, a well-studied multiple sclerosis model (MS). A total of 1 × 106 BMSCs, ASCs, or SVF cells were administered intraperitoneally concomitantly with the induction of disease. Mice were monitored daily for clinical signs of disease by three independent, blinded investigators and rated on a scale of 0 to 5. Spinal cords were obtained after euthanasia at day 30 and processed for histological staining using luxol fast blue, toluidine blue, and hematoxylin and eosin to measure myelin and infiltrating immune cells. Blood was collected from mice at day 30 and analyzed by enzyme-linked immunosorbent assay to measure serum levels of inflammatory cytokines.ResultsThe data indicate that intraperitoneal administration of all cell types significantly ameliorates the severity of disease. Furthermore, the data also demonstrate, for the first time, that the SVF was as effective as the more commonly cultured BMSCs and ASCs in an MS model. All cell therapies also demonstrated a similar reduction in tissue damage, inflammatory infiltrates, and sera levels of IFNγ and IL-12. While IFNγ levels were reduced to comparable levels between treatment groups, levels of IL-12 were significantly lower in SVF-treated than BMSC-treated or ASC-treated mice.ConclusionsBased on these data, it is evident that SVF cells have relevant therapeutic potential in an animal model of chronic MS and might represent a valuable tool for stem cell-based therapy in chronic inflammatory disease of the central nervous system. SVF offers advantages of direct and rapid isolation procedure in a xenobiotic-free environment.

Helminth Therapy for MS.

In the last 50 years, environmental factors such as helminth infections have been proposed to explain why autoimmunity is less prevalent in the developing world; this proposal has been termed the hygiene or old friends hypothesis. The epidemiology of MS shows an inverse correlation with helminth infections. Positive effects of helminths in animal models of MS and observational studies in people with MS naturally infected with helminths suggest that those organisms can act as immune regulators and led to clinical trials of helminth therapy. The goal of helminth therapy is to introduce parasitic organisms into people with MS in a controlled and predictable fashion, and to prevent immune-mediated disease without increasing the risk of pathology with high parasite load. This chapter focuses on intestinal worms as they are the current choice as a therapeutic strategy in a number of autoimmune diseases, including MS. Here we review current data regarding the rationale and the current state of research in the field of helminth therapies in MS.

Amelioration of autoimmune neuroinflammation by the fusion molecule Fn14·TRAIL

BackgroundMultiple sclerosis (MS) is a, T cell-mediated autoimmune disease, the management of which remains challenging. The recently described fusion protein, Fn14·TRAIL, combining the extracellular domain of Fn14 (capable of blocking the pro-inflammatory TWEAK ligand) fused to the extracellular domain of the TRAIL ligand (capable of sending apoptotic signals through its receptors on activated inflammatory cells) was designed to modulate the immune system as an anti-inflammatory agent. The present study explores the efficacy of this purified protein as an anti-inflammatory agent, using the animal model of MS - experimental autoimmune encephalomyelitis (EAE).MethodsEAE was induced by myelin oligodendrocyte glycoprotein (MOG). Fn14·TRAIL or vehicle were injected daily for 4 to 16 days, at different time points after disease induction. Animals were examined daily and evaluated for EAE clinical signs. Lymphocytes were analyzed for ex vivo re-stimulation, cytokine secretion, transcription factor expression and subtype cell analysis. Spinal cords were checked for inflammatory foci. The Mann- Whitney rank sum test, Student’s t-test or ANOVA were used for statistical analysis.ResultsSignificant improvement of EAE in the group treated with Fn14·TRAIL was noted from day 6 of disease onset and lasted until the end of follow-up (day 40 from disease induction), even in animals treated for 4 days only. Clinical improvement was linked to decreased lymphocyte infiltrates in the central nervous system (CNS) and to decreased Th1 and Th17 responses and to increased number of T- regulatory in the treated mice. No liver or kidney toxicity was evident. In vitro assays established the ability of Fn14·TRAIL to induce apoptosis of T cell lines expressing TRAIL receptors and TWEAK.ConclusionsIn this study we established the potency of Fn14·TRAIL, a unique fusion protein combining two potentially functional domains, in inhibiting the clinical course of EAE, even when given for a short time, without apparent toxicity. These findings make Fn14·TRAIL a highly promising agent to be used for targeted amelioration of neuro-inflammatory processes, as well as other autoimmune pathologies.

Changes in nociceptive sensitivity and object recognition in experimental autoimmune encephalomyelitis (EAE)

Multiple sclerosis is associated with a high incidence of depression, cognitive impairments and neuropathic pain. Previously, we demonstrated that tactile allodynia is present at disease onset in an animal model of MS, experimental autoimmune encephalomyelitis (EAE). We have now monitored changes in object recognition in mice with EAE to determine if altered nociceptive sensitivity is also associated with behavioral signs indicative of cognitive impairment in this model. At the onset of clinical signs, mice with EAE showed impairments in the novel object recognition (NOR) assay, indicative of deficits in cognitive functioning early in the disease course. At the spinal level, we found increased gene expression for the cytokines IL-1β, IL-6 and the glutamate transporter EAAT-2 that coincide with increased nociceptive sensitivity and deficits in object recognition. Increased levels of EAAT-2 mRNA appear to be a response to perturbed protein levels of the transporter as we found a loss of EAAT-2 protein levels in the spinal cord of EAE mice. To determine if changes in the levels of EAAT-2 were responsible for the observed changes in nociceptive sensitivity and cognitive deficits, we treated EAE mice with the β-lactam antibiotic ceftriaxone, an agent known to increase glutamate transporter levels in vivo. Ceftriaxone prevented tactile hypersensitivity and normalized performance in the NOR assay in EAE mice. These findings highlight the important interrelationship between pain and cognitive function in the disease and suggest that targeting spinally mediated pain hypersensitivity is a novel therapeutic avenue to treat impairments in other higher order cortical processes.

Intranasal Delivery of Neural Stem Cells: A CNS-specific, Non-invasive Cell-based Therapy for Experimental Autoimmune Encephalomyelitis

The therapeutic potential of adult neural stem cells (aNSCs) has been shown in EAE, an animal model of MS, administered by either i.c.v. or i.v. injection. However, i.c.v. is an invasive approach, while the i.v. route of aNSCs is associated with a non-specific immune suppression in the periphery. Here we demonstrate that intranasal (i.n.) delivery of fluorescently labeled aNSCs resulted in their appearance in the olfactory bulb, cortex, hippocampus, striatum, brainstem, and spinal cord. These cells induce functional recovery from ongoing EAE similar to that achieved with i.v. injected aNSCs, with comparable anti-inflammatory and remeylination effects in CNS inflammatory foci. Importantly, unlike the peripheral immune suppression brought about by i.v. NSCs, intranasal delivery did not influence peripheral immune responses. We conclude that aNSCs can be reliably delivered to the CNS via the nasal route to induce functional recovery and confer immunomodulation and remyelination in EAE. Intranasal administration of NSCs provides a highly promising, noninvasive and CNS-specific alternative to current cell-based approaches in treating EAE.

Elevation of Sema4A Implicates Th Cell Skewing and the Efficacy of IFN-β Therapy in Multiple Sclerosis

Multiple sclerosis (MS) is a demyelinating autoimmune disease of the CNS and a leading cause of lasting neurologic disabilities in young adults. Although the precise mechanism remains incompletely understood, Ag presentation and subsequent myelin-reactive CD4(+) T cell activation/differentiation are essential for the pathogenesis of MS. Although semaphorins were initially identified as axon guidance cues during neural development, several semaphorins are crucially involved in various phases of immune responses. Sema4A is one of the membrane-type class IV semaphorins, which we originally identified from the cDNA library of dendritic cell (DC). Sema4A plays critical roles in T cell activation and Th1 differentiation during the course of experimental autoimmune encephalomyelitis, an animal model of MS; however, its pathological involvement in human MS has not been determined. In this study, we report that Sema4A is increased in the sera of patients with MS. The expression of Sema4A is increased on DCs in MS patients and shed from these cells in a metalloproteinase-dependent manner. DC-derived Sema4A is not only critical for Th1 but also for Th17 cell differentiation, and MS patients with high Sema4A levels exhibit Th17 skewing. Furthermore, patients with high Sema4A levels have more severe disabilities and are unresponsive to IFN-β treatment. Taken together, our results suggest that Sema4A is involved in the pathogenesis of MS by promoting Th17 skewing.

Repulsive Guidance Molecule-a and Demyelination: Implications for Multiple Sclerosis

Drug development for neurodegenerative and neuroinflammatory diseases such as multiple sclerosis and traumatic brain injury is challenging. One promising strategy is to target a molecule with multiple biological actions affecting divergent pathophysiological disease phases simultaneously since these diseases arise from multiple pathological phases. In recent years, we pursued this strategy with a focus on multiple sclerosis and spinal cord injury and found that repulsive guidance molecule-a (RGMa) inhibits regeneration of injured CNS axons following spinal cord injury. We also found that RGMa enhances CD4(+) T cell activation facilitating CNS demyelination in an animal model of MS, mouse experimental autoimmune encephalomyelitis (EAE), which supports the idea that RGMa has distinct pathological actions. The multiple functions of RGMa in the CNS and the immune system would provide a therapeutic opportunity to concurrently block the autoimmune reactions and axon injury in neurodegenerative and neuroinflammatory diseases. In this article, we introduce the therapeutic potential of targeting RGMa as a novel intervention for MS and spinal cord injury.

A Missense Mutation in Myelin Oligodendrocyte Glycoprotein as a Cause of Familial Narcolepsy with Cataplexy

Narcolepsy is a rare sleep disorder characterized by excessive daytime sleepiness and cataplexy. Familial narcolepsy accounts for less than 10% of all narcolepsy cases. However, documented multiplex families are very rare and causative mutations have not been identified to date. To identify a causative mutation in familial narcolepsy, we performed linkage analysis in the largest ever reported family, which has 12 affected members, and sequenced coding regions of the genome (exome sequencing) of three affected members with narcolepsy and cataplexy. We successfully mapped a candidate locus on chromosomal region 6p22.1 (LOD score ¼ 3.85) by linkage analysis. Exome sequencing identified a missense mutation in the second exon of MOG within the linkage region. A c.398C>G mutation was present in all affected family members but absent in unaffected members and 775 unrelated control subjects. Transient expression of mutant myelin oligodendrocyte glycoprotein (MOG) in mouse oligodendrocytes showed abnormal subcellular localization, suggesting an altered function of the mutant MOG. MOG has recently been linked to various neuropsychiatric disorders and is considered as a key autoantigen in multiple sclerosis and in its animal model, experimental autoimmune encephalitis. Our finding of a pathogenic MOG mutation highlights a major role for myelin and oligodendrocytes in narcolepsy and further emphasizes glial involvement in neurodegeneration and neurobehavioral disorders. [corrected].