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Human T-lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a neurological disease that results from an interaction of retroviral infection and immune activation. In this study, five doses (1 mg/kg) of humanized anti-Tac antibody were administered to 9 HAM/TSP patients at weeks 0, 2, 6, 10, and 14. Preliminary immunological studies on HAM/TSP patients treated with humanized anti-Tac indicate that there is a selective down-regulation of activated T cells and a decrease in the HTLV-I viral load in peripheral blood lymphocytes, most likely through the selective removal of HTLV-I-infected, activated CD4+ lymphocytes.

Patients who develop relapsing-remitting multiple sclerosis (MS) present with a first clinical demyelinating event. In this double-blind, multicentre, randomised, phase 3 study we investigated the effect of oral cladribine on conversion to clinically definite MS in patients with a first clinical demyelinating event, when given at the same doses shown to be effective in relapsing-remitting MS.Between Oct 21, 2008, and Oct 11, 2010, we recruited patients aged 18-55 years, inclusive, from 160 hospitals, private clinics, or treatment centres in 34 countries. Eligible patients had a first clinical demyelinating event within 75 days before screening, at least two clinically silent lesions of at least 3 mm on a T2-weighted brain MRI scan, and an Expanded Disability Status Scale score of 5.0 or lower. Patients with a first clinical demyelinating event ≤75 days before screening were randomly assigned (1:1:1) to receive cladribine tablets at cumulative doses of 5.25 mg/kg or 3.5 mg/kg or placebo. Randomisation was done with a central web-based randomisation system and was stratified by geographic region. Masking was maintained using a two-physician model. The primary endpoint of this 96-week study was time to conversion to clinically definite MS according to the Poser criteria. This study is registered with ClinicalTrials.gov, number NCT00725985.Of 903 participants assessed for eligibility, 616 patients received cladribine 5.25 mg/kg (n=204), cladribine 3.5 mg/kg (n=206), or placebo (n=206). At trial termination on Oct 25, 2011, cladribine was associated with a risk reduction versus placebo for time to conversion to clinically definite MS (hazard ratio [HR] for 5.25 mg/kg=0.38, 95% CI 0.25-0.58, p0.0001; HR for 3.5 mg/kg=0.33, 0.21-0.51, p0.0001). Adverse events were reported in 165 (81%) patients in the cladribine 5.25 mg/kg group, 168 (82%) patients in the cladribine 3.5 mg/kg group, and 162 (79%) patients in the placebo group. We noted no increase in risk of adverse events with active treatment versus placebo apart from lymphopenia, which was a severe event in 10 (5%) patients in the 5.25 mg/kg group and four (2%) patients in the 3.5 mg/kg group.Both doses of cladribine significantly delayed MS diagnosis compared with placebo. The safety profile of cladribine was similar to that noted in a trial in patients with relapsing-remitting MS. Further research could clarify the potential effects of oral cladribine treatment in the early stages of MS.Merck Serono SA Geneva, a subsidiary of Merck KGaA, Darmstadt, Germany.

Multiple sclerosis (MS) affects an estimated 300,000 individuals in the United States. No cure exists and although there is a lack of consensus on management, strategies to modify disease course are available. These strategies involve initiating disease-modifying therapies that have been found to slow disease progression and prevent disability symptoms, thereby improving function for MS patients. The overall goal of early disease management is to intervene prior to irreversible neuronal destruction in order to delay disability progression and improve quality of life. Maintaining a lower level of disability for a longer period of time postpones and ultimately attempts to prevent reaching a level of immobility and irreversible disability. However, due to the complex nature of disease and its unique, individual patient course, no patient can be treated alike and no patient responds to therapy similarly. Therefore, MS research is continuous in its evolution of therapeutic development, focusing on neuroprotective effects and agents with distinctive mechanisms of action allowing for unique safety and efficacy profiles. Investigations include novel oral agents and monoclonal antibodies. Many of the approved agents also are continually being investigated in order to evaluate comparative data, the most appropriate means of implementing subsequent therapy upon failure, responsiveness to therapeutic agent when switched, and long-term safety and efficacy. This multimedia webcast educational activity will cover the current state of MS science, current therapies in MS, emerging treatments in clinical trials for MS as well as differences between physicians in diagnosis and management of MS and their evolving practices.

Objective To examine associations between multitasking ability defined by performance on a complex task integrating multiple cognitive domains and vocational functioning in multiple sclerosis (MS). Design Survey data collection. Setting Laboratory with referrals from an outpatient clinic. Participants Community-dwelling individuals with MS (N=30) referred between October 2011 and June 2012. Interventions Not applicable. Main Outcome Measures The modified Six Elements Test (SET) to measure multitasking ability, Fatigue Severity Scale to measure fatigue, several neuropsychological measures of executive functioning, and vocational status. Results Among the sample, 60% of individuals have reduced their work hours because of MS symptoms (cutback employment group) and 40% had maintained their work hours. Among both groups, SET performance was significantly associated with performance on several measures of neuropsychological functioning. Individuals in the cutback employment group demonstrated significantly worse overall performance on the SET ( P =.041). Logistic regression was used to evaluate associations between SET performance and vocational status, while accounting for neuropsychological performance and fatigue. The overall model was significant (χ 2 3 =8.65, P =.032), with fatigue [Exp(B)=.83, P =.01] and multitasking ability [Exp(B)=.60, P =.043] retained as significant predictors. Conclusions Multitasking ability may play an important role in performance at work for individuals with MS. Given that multitasking was associated with vocational functioning, future efforts should assess the usefulness of incorporating multitasking ability into rehabilitation planning.

Introduction L’efficacite est parfois comparee via les risques relatifs. Mais les resultats peuvent etre influences par des differences entre les populations. Le NNT est une approche complementaire pour la comparaison entre traitements. Objectifs Comparer le NNT pour eviter une poussee ou l’aggravation du handicap confirmee a 12 semaines dans les essais avec le teriflunomide ou le dimethyl fumarate (DMF). Patients et methodes Dans cette analyse post-hoc, les NNT sont determines sur la base des donnees des etudes avec le teriflunomide 14 mg (TEMSO, NCT00134563 ; TOWER, NCT00751881 ) ou le DMF (DEFINE, NCT00420212 ; CONFIRM, NCT00451451 ). Ils sont obtenus en calculant l’inverse du risque absolu, differentiel entre les groupes de traitement actif et le placebo. Resultats Le NNT pour eviter une poussee est comparable entre les etudes (TEMSO : 5,9 ; TOWER : 5,6 ; DEFINE : 5,3 ; CONFIRM : 5,6). Le risque d’aggravation du handicap est significativement reduit dans TEMSO, TOWER et DEFINE mais pas dans CONFIRM. Les NNT respectifs sont de 13,8 ; 17,4 ; 10,8 et 30,2. Le risque de poussee entrainant l’hospitalisation est significativement reduit dans TEMSO et TOWER mais pas dans DEFINE et CONFIRM. Les NNT sont plus faibles dans TEMSO et TOWER (12,5 ; 20) que dans DEFINE et CONFIRM (50 ; 50). Discussion Le teriflunomide et le DMF sont deux nouveaux traitements per os. Les quatre etudes pivot (TEMSO, TOWER, DEFINE et CONFIRM) ont montre une efficacite sur la prevention des poussees et le teriflunomide est le seul a avoir demontre son efficacite sur le handicap dans deux etudes. Les NNT pour prevenir l’aggravation du handicap sont comparables pour les etudes avec le teriflunomide et DEFINE, et plus eleve pour CONFIRM. Conclusion En utilisant le NNT, l’effet sur les poussees est comparable entre le teriflunomide et le DMF. Informations complementaires Etude realisee avec le soutien de Genzyme, une filiale de Sanofi.

Diffusion tensor imaging (DTI) of the human optic nerve presents challenges from geometric distortion in echo planar imaging (EPI) caused by magnetic field inhomogeneity and partial volume artifacts caused by confounding signals from surrounding fat and cerebrospinal fluid (CSF). A protocol for human optic nerve DTI was developed with geometric distortion correction and suppression of fat and CSF signals. This protocol was modified from a conventional DTI protocol to acquire images and field maps covering the whole brain including contiguous slices of the optic nerves. The technique was applied to healthy volunteers and multiple sclerosis (MS) patients with and without history of unilateral optic neuritis (ON). DTI measures of the optic nerves before and after distortion correction were compared. Means and standard deviations of these measures from different cohorts were reported.

To assess the risk of developing multiple sclerosis (MS) after optic neuritis and the factors predictive of high and low risk.Subjects in the Optic Neuritis Treatment Trial, who were enrolled between July 1, 1988, and June 30, 1991, were followed up prospectively for 15 years, with the final examination in 2006.Neurologic and ophthalmologic examinations at 13 clinical sites.Three hundred eighty-nine subjects with acute optic neuritis.Development of MS and neurologic disability assessment.The cumulative probability of developing MS by 15 years after onset of optic neuritis was 50% (95% confidence interval, 44%-56%) and strongly related to presence of lesions on a baseline non-contrast-enhanced magnetic resonance imaging (MRI) of the brain. Twenty-five percent of patients with no lesions on baseline brain MRI developed MS during follow-up compared with 72% of patients with 1 or more lesions. After 10 years, the risk of developing MS was very low for patients without baseline lesions but remained substantial for those with lesions. Among patients without lesions on MRI, baseline factors associated with a substantially lower risk for MS included male sex, optic disc swelling, and certain atypical features of optic neuritis.The presence of brain MRI abnormalities at the time of an optic neuritis attack is a strong predictor of the 15-year risk of MS. In the absence of MRI-detected lesions, male sex, optic disc swelling, and atypical clinical features of optic neuritis are associated with a low likelihood of developing MS. This natural history information is important when considering prophylactic treatment for MS at the time of a first acute onset of optic neuritis.

Geometric distortion induced by magnetic field inhomogeneity is an intrinsic problem for echo-planar imaging (EPI). Most of the existing correction techniques require separate field map scans in addition to the measurement scans. The effectiveness of these methods requires consistent patient position between these scans, thus inter-scan patient motions degrade the correction results. We propose a new method for geometric distortion correction in EPI that derives the field maps from the measurements themselves, without the need of additional field map scans. This method takes advantage of parallel imaging and k-space trajectory modification to generate multiple images for field mapping from a single measurement needed. The derived field maps are retrospectively applied to the measurements. The proposed method has been successfully demonstrated on combined k- space data that simulate the data from the modified k-space trajectories. These data were acquired from a phantom and a human brain.

The roles of tumor necrosis factor (TNF alpha) and reactive nitrogen intermediates (RNI) as effectors of macrophage-mediated tumor cell killing were investigated in a variety of tumor cell lines. Three TNF alpha-sensitive tumor targets were also susceptible to resting bone-marrow-derived mononuclear phagocytes (BMMP). This macrophage lytic activity was markedly diminished or even abolished by anti-TNF alpha, indicating that TNF alpha is the major effector of macrophage-mediated killing of these targets. The other 21 tumor cell lines examined were resistant to TNF alpha but, in their large majority, were more or less susceptible to killing by interferon gamma (IFN gamma)- and Corynebacterium parvum (CP)-activated BMMP. Among the various analogues of L-arginine used to assess the role of L-arginine-derived RNI as mediators of macrophage tumoricidal activity, NG-monomethyl-L-arginine (NMMA) was most efficient in suppressing RNI secretion by activated macrophages. In some macrophage tumor-cell combinations, NMMA inhibited both the generation of RNI and the expression of tumoricidal activity in a dose-dependent manner, suggesting a central role for RNI as effectors. In other combinations, NMMA in concentrations that abolished secretion of RNI either affected tumor-cell killing only after its induction by IFN gamma, or not at all. The findings not only support the thesis that macrophages posses various means of coping with tumor cells but also suggest that the mechanism becoming operative is determined predominantly by the pathway of macrophage activation and the properties of the tumor-cell type.

The complex processes that determine the outcome of the interaction of tumor and host were explored in the operationally simple and reproducible rat D-12 ascites tumor model. Animals exhibit weak spontaneous resistance against this tumor that is not augmented by repeated inoculation, by various routes, of irradiated syngeneic D-12 tumor cells, but considerably enhanced after local administration of heat-killed Corynebacterium parvum (CP) or Listeria monocytogenes (LM) organisms. Inoculation of conventional or monoclonal anti-rat IFN gamma antibodies into the same compartment did not affect spontaneous tumor resistance, but largely abrogated the tumor-protective effect triggered by CP or LM. Our findings support the concept that IFN gamma, produced by T cells in the course of the specific immune response raised against immunogenic micro-organisms, is able to enhance and to maintain local tumor resistance and thus to strengthen the capacity of the host to cope with a non-immunogenic tumor.

IntroductionTeriflunomide, approved for the treatment of relapsing-remitting multiple sclerosis, has a well-characterized safety profile based on individual clinical studies. We report pooled safety and tolerability data from four, double-blind, placebo-controlled trials of teriflunomide. Post-approval updates on hair thinning and pregnancy outcomes, sometimes concerns for patients initiating teriflunomide, are reported.MethodsData were pooled from phase 2 (NCT01487096) and phase 3 TEMSO (NCT00134563), TOWER (NCT00751881), and TOPIC (NCT00622700) studies. Patients were randomized to receive teriflunomide 14 mg, 7 mg, or placebo. Safety analyses were performed for all patients exposed to teriflunomide.ResultsThe pooled dataset included 3044 patients. Commonly reported adverse events (AEs) were in accordance with individual clinical studies, most being transient and mild-to-moderate in intensity. Incidence of hepatic AEs was higher in teriflunomide groups; however, serious hepatic AEs were similar across groups (∼2–3%). Hair thinning was higher in teriflunomide than placebo groups, but typically resolved on treatment without intervention and led to discontinuation in ConclusionsThese data from >6800 patient-years of teriflunomide exposure were consistent with individual studies and no new, unexpected safety signals were observed. (Study supported by Genzyme, a Sanofi company).

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.

To assess the safety, tolerability and pharmacokinetics of an interleukin (IL)-12/23 monoclonal antibody (mAb) in subjects with a relapsing form of multiple sclerosis (MS).A phase I, double-blind, placebo-controlled, sequential dose escalation study was conducted in 20 subjects with MS. Subjects were randomized (4:1) to receive a single subcutaneous injection of either IL-12/23 mAb (0.3, 0.75, 1.5, and 3.0 mg/kg) or placebo. Clinical and laboratory evaluations were performed through 16 weeks following administration.IL-12/23 mAb was well tolerated in this study. Adverse events were generally mild or moderate, with no apparent dose-related trends. One subject with a family history of breast cancer was diagnosed during the study with breast cancer 21 days after IL-12/23 mAb administration. There were no significant changes in laboratory indicators of systemic or neurotoxicity. There was a large degree of variability in T2 lesion volume and total number of gadolinium-positive lesions, both unaffected by dose escalation. Three relapses of MS occurred in two placebo-treated subjects. Over the range of single doses studied, the median Tmax ranged from 9.0 to 16.5 days, and the median T1/2 ranged from 20.2 to 30.9 days.Single subcutaneous administrations of IL-12/23 mAb in this first study of relapsing MS were generally well tolerated. Safety of the agent will need to be tested in a study of longer duration and involving a larger cohort of subjects.

Human T-lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is an immune-mediated inflammatory disorder of the central nervous system. Immune activation in the host, which results from high levels of persistent antigenic stimulation and from transactivation of host immunoregulatory genes by HTLV-I, appears important in the pathogenesis of HAM/TSP. In a single-center, open-label trial, 12 patients with HAM/TSP were treated with doses of interferon-beta1a of up to 60mug twice weekly, based on its antiviral and immunomodulatory effects. Primary end points were immunological and virological measures that are potential biomarkers for HAM/TSP. Interferon-beta1a therapy reduced the HTLV-I tax messenger RNA load and the frequency of potentially pathogenic HTLV-I-specific CD8(+) cells. The HTLV-I proviral DNA load remained unchanged. Spontaneous lymphoproliferation, a marker of T-cell activation in HAM/TSP, also was reduced. Some measures of motor function were improved, and no significant clinical progression occurred during therapy. These results indicate that interferon-beta1a may beneficially affect the immune mechanisms central to the pathogenesis of HAM/TSP.

The importance of β-chemokines (or CC chemokine ligands – CCL) in the development of inflammatory lesions in the central nervous system of patients with multiple sclerosis and rodents with experimental allergic encephalomyelitis is strongly supported by descriptive studies and experimental models. Our recent genetic scans in families identified haplotypes in the genes of CCL2, CCL3 and CCL11-CCL8-CCL13 which showed association with multiple sclerosis. Complementing the genetic associations, we also detected a distinct regional expression regulation for CCL2, CCL7 and CCL8 in correlation with chronic inflammation in multiple sclerosis brains. These observations are in consensus with previous studies, and add new data to support the involvement of CCL2, CCL7, CCL8 and CCL3 in the development of inflammatory demyelination. Along with our own data, here we review the literature implicating CCLs and their receptors (CCRs) in multiple sclerosis and experimental allergic encephalomyelitis. The survey reflects that the field is in a rapid expansion, and highlights some of the pathways which might be suitable to pharmaceutical interventions.

The 77C→ G polymorphism in exon 4 of CD45 or protein tyrosine phosphatase receptor-type C (PTPRC) has been investigated in families with multiple sclerosis (MS) and in several cohorts of sporadic patients and controls, however, with conflicting results. To better understand the role of this functionally important polymorphism in MS, we investigated the transmission of the ‘G’ allele from unaffected parents to their affected children in 176 families by using linkage and association statistics. The ‘G’ allele was transmitted to the affected offspring in five of the seven pedigrees carrying this allele and the TRANSMIT program detected association with a P -0.0342. We conclude that the 77C→ G PTPRC polymorphism is present and preferentially transmitted in a small subgroup (< 5%) of MS families, which may only be detected with complementary methods of analysis.

Linkage studies in multiple sclerosis (MS) identified several susceptibility loci. One of these regions includes chromosome 17q11 where a meta-analysis of data from three genome scans suggested linkage. This region encodes a cluster of genes for beta-chemokines or CC chemokine ligands (CCLs), which may be involved in the development of MS lesions. Here we aimed to test if CCL alleles and haplotypes are associated with MS. Using methods of linkage and association, we observed deviations from the expected 50% transmission of haplotypes from unaffected parents to their affected children at CCL2, CCL11-CCL8-CCL13 and CCL3 within the investigated 1.85 MB chromosomal segment. Analyses of the linkage disequilibrium map support that variants with possible relevance to MS can be located within these subregions. Identification of MS associated CCL variants may have direct clinical significance, as it can lead to the design of small competitive antagonists of these molecules with beneficial effects in the treatment of patients with early and active disease.

Hypothesis A mitochondrial mechanism contributes to neurodegeneration in multiple sclerosis (MS). Genetic variants of Complex I genes may influence the nature of tissue response to inflammation in the central nervous system (CNS). Background Complex I is encoded by seven mitochondrial and 38 nuclear genes. Many of the nuclear genes colocalize with regions where full genome scans detected linkage in MS. Previous studies revealed an association between variants of mitochondrial (mt)DNA encoded subunits of Complex I and MS. Biochemical studies suggested a functional involvement of Complex I in the degenerative processes downstream to inflammatory injury in the CNS. Methods Patients with all MS phenotypes were included. DNA specimens of affected sib pair, trio and multiplex families were studied. Single nucleotide polymorphisms (SNP) were determined by using the Taqman assay. The association of MS with nuclear DNA encoded alleles and haplotypes of Complex I was tested by the pedigree disequilibrium test (PDT) and by the transmit program in the families. Haplotypes were further investigated by using ldmax (GOLD). The association of mtDNA encoded variants with MS was tested by the Fisher's Exact Test. Results The previously identified MS-associated mtDNA variants and haplotypes were not increased in mothers as compared to fathers in these families. However, an association of all clinical phenotypes with haplotypes within NDUFS5 (1p34.2-p33), NDUFS7 (19p13) and NDUFA7 (19p13) was detected. The inclusion of families with primary progressive (PP)-MS phenotype did not modify the outcome and, as a subgroup alone, did not have a sufficient size to draw conclusion regarding phenotype specific associations. Conclusions SNP haplotypes within Complex I genes are associated with MS. Further studies are needed to refine the identification of disease relevant variants nearby or within these haplotypes. Molecular and functional properties of Complex I subunits may offer novel explanations to better understand the relationship between inflammation and neurodegeneration.

The human T-cell lymphotropic virus type I (HTLV-I) is associated with a chronic, progressive neurological disease known as HTLV-I‐associated myelopathy/tropical spastic paraparesis. Screening for HTLV-I involves the detection of virus-specific serum antibodies by EIA and confirmation by Western blot. HTLV-I/II seroindeterminate Western blot patterns have been described worldwide. However, the significance of this blot pattern is unclear. We identified 8 patients with neurological disease and an HTLV-I/II seroindeterminate Western blot pattern, none of whom demonstrated increased spontaneous proliferation and HTLVI‐specific cytotoxic T lymphocyte activity. However, HTLV-I tax sequence was amplified from the peripheral blood lymphocytes of 4 of them. These data suggest that patients with chronic progressive neurological disease and HTLV-I/II Western blot seroindeterminate reactivity may harbor either defective HTLV-I, novel retrovirus with partial homology to HTLV-I, or HTLVI in low copy number. The human T-cell lymphotropic virus type I (HTLV-I) is endemic in several regions of the world; there are clusters of high prevalence in the Caribbean, southern Japan, equatorial Africa, South America, and Iran [1]. Long established as the causative agent of both adult T-cell leukemia/lymphoma [2] and an inflammatory neurological disease termed HTLV-I‐associated myelopathy/tropical spastic paraparesis (HAM/TSP) [3], HTLV-I has more recently been implicated in other inflammatory diseases in a subset of patients, including HTLVI‐associated uveitis and infectious dermatitis [4, 5]. Although

Objective: To evaluate characteristics related to 2-year relapse rate for the TOP MS study completer cohort. Background The Therapy Optimization in MS Study (TOP MS) is a prospective, open-label parallel group study of patients who are being treated with disease-modifying therapies (DMT). All of the DMT have been shown to effectively reduce relapse rates. Design/Methods: Potential participants for TOP MS were identified at specialty pharmacies with medication therapy management programs. Signed informed consent forms were returned to the pharmacies, and study enrollment produced log-on instructions for the study website. At baseline and monthly intervals over 24 months, enrolled participants receive reminders to respond to surveys. Self-reported responses are entered directly into the study database. Results: The TOP MS completer cohort consists of 531 subjects with a mean duration of disease of 11.1+9.2 years. Nearly 90% of participants met the criteria for relapsing forms of MS. The 2-year relapse rate for this cohort was 35.3%. Glatiramer acetate was being used by 50.8%, and 49.2% were using interferon-beta at study enrollment. Over two years therapy change occurred for 50 patients (9.4%) and was significantly associated with the number of confirmed relapses (chi 2 = 13.9, df = 2, p = .001). While mean duration of current DMT treatment was 4.3+4.1 years, shorter duration was correlated with more confirmed relapses (p = .004). One-third of the completer cohort had used other DMT prior to starting their current therapy (mean number of previous therapies: 1.3) and number of previous DMT was positively correlated with number of confirmed relapses (p = .001). Disease duration and enrollment DMT were not associated with relapses. Conclusions: The two-year relapse rate in the completer cohort of the TOP MS Study was influenced by change in DMT therapy, time on DMT treatment, and number of previous DMT treatments. Supported by: Teva Pharmaceuticals. Disclosure: Dr. Coyle has received personal compensation for activities with Acorda Therapeutics, Avanir Pharmaceuticals, Bayer Pharmaceuticals Corporation, Biogen Idec, Genzyme Corporation, Novartis, Questcor, Roche Diagnostics Corporation, Sanofi-Aventis Pharmaceuticals, Inc., and Teva Neuroscience. Dr. Coyle has received personal compensation in an editorial capacity for NEURA. Dr. Coyle has received research support from Serono, Inc., Novartis, and Sanofi-Aventis Pharmaceuticals, Inc. Dr. Zwibel has received personal compensation for activities with Serono, Inc., and Teva Neuroscience. Dr. Zwibel has received research support from Teva Neuroscience. Dr. Cohen has received research support from Biogen Idec. Dr. Leist has received personal compensation for activities with EMD Serono, Teva Neuroscience and Bayer.Dr. Leist has received research support from Bayer, EMD Serono and Teva Neuroscience. Dr. Tullman has received personal compensation for activities with Acorda Therapeutics, Biogen Idec, Serono Inc., Novartis, Pfizer Inc., Teva Neuroscience, Sanofi-Aventis Pharmaceuticals, Abbott Labortories, Inc. and Vaccinex as a consultant and/or speaker. Dr. Tullman has received research support from Acorda Therapeutics, BioMS, Novartis, and Genentech, Inc. Dr. Markowitz has received personal compensation for activities with Bayer, Teva, EMD-Serono, Biogen Idec, and Eli Lilly. Dr. Hughes has received research support from Teva Neuroscience, Berlex Labratories and Genentech, Inc..

INTRODUCTION In multiple sclerosis (MS), both inflammatory demyelination and axonal loss in gray and white matter are observed early, even presymptomatically [1]. These losses lead to central nervous system (CNS) atrophy [2] and functional disability [3]. Standard outcome measures of relapse rate and magnetic resonance imaging (MRI) lesion load neither correlate well with functional decline nor reflect CNS neurodegeneration [4-5]. Optical coherence tomography (OCT) is under evaluation as an outcome measure of axonal degeneration for use in clinical trials and practice [6]. OCT is a noninvasive technique that uses infrared laser light to create cross-sectional images of the retina; this technique provides quantitative analysis of the retinal nerve fiber layer (RNFL) thickness (RNFLt), macular thickness, and macular volume. The mechanism and reliability of OCT are well described [7]. OCT detects changes in the RNFL, which contains unmyelinated axons subject to damage in glaucomatous and ischemic optic neuropathies [8]; more recently, OCT has been applied to optic neuritis (ON) [9]. Short-term follow-up studies after acute ON demonstrate an acute decrement of the peripapillary RNFLt that plateaus by 3 to 6 months [10]. OCT also detects changes in the macula after ON [11]. The calculated macular volume includes the inner macular thickness, which contains the retinal ganglion cells presumably affected after ON by a dying-back axonopathy, as seen in animal models [12]. Alternatively or additionally, primary neuronal degeneration may affect the retinal ganglion cells. OCT has been shown to correlate with functional optic nerve measures such as multifocal visual-evoked potentials [13] and low-contrast visual acuity (LCVA) [14]. As one of the longer white matter tracts in the CNS, the optic nerve is likely to show the effects of both acute inflammation and neurodegeneration early in MS. ON is the presenting symptom of MS in 25 to 50 percent of cases and eventually affects up to 80 percent of patients during the illness, causing significant morbidity [15]. RNFLt in subjects with MS without a history of ON is lower than in control subjects, indicating subclinical axonal loss [9,16]. Thus, OCT is an attractive candidate measure for capturing early changes in MS. The corticospinal tract is another long CNS white matter tract frequently affected in MS. Like the optic nerve, it is subject both to acute inflammation in the form of transverse myelitis and to neurodegeneration as seen by increasing gait dysfunction, MRI atrophy [2], and recently, gray matter demyelination on high-field MRI [17]. The Expanded Disability Status Scale (EDSS) is a standard measure of gait dysfunction and may correlate with other long tract signs, including RNFL loss and cognitive dysfunction [18], although it poorly predicts longterm disability [19] and its validity in relapsing-remitting MS (RRMS) has been recently challenged [20]. OCT may provide a more sensitive research and clinically applicable measure to capture neurodegeneration in MS. Most studies using OCT have not accounted for the effects of disease-modifying therapies (DMTs) on the RNFL, which may be significant if similar to DMT effects in early MS [21]. We studied OCT in subjects with MS of varying disease durations and with no exposure to DMTs to test the hypothesis that in untreated MS, RNFLt loss occurs both over the course of disease and in parallel with corticospinal tract dysfunction reflecting CNS neurodegeneration. METHODS Subjects The institutional review board approved all study procedures, and subjects gave their written informed consent. We recruited subjects over 18 years of age from the Thomas Jefferson University Comprehensive MS Center (Philadelphia, Pennsylvania) and the Neuro-Ophthalmology Service at Wills Eye Institute (Philadelphia, Pennsylvania) with no exposure to DMT. Subjects met revised McDonald criteria for MS [22]. …

The effects of sheep anti-murine recombinant tumor necrosis factor-alpha (TNF-alpha) on resistance to Listeria monocytogenes infection were studied in T cell-deficient nu/nu mice. The sheep anti-TNF-alpha antibody preparation was specific for TNF since it neutralized 300 U of recombinant murine TNF-alpha in vitro at a dilution of up to 1/1,000 but did not neutralize 32 U of interferon (IFN)-alpha, -beta or 32 U of IFN-gamma in vitro at a 1/20 dilution. When tested in vivo in sublethally Listeria-infected nu/nu or T cell-competent C57BL/6 or ICR mice, a single treatment of 0.2 ml anti-TNF-alpha given intraperitoneally on either day -1,0 or +1 resulted in the death of mice by day 5-7 due to the uncontrolled growth of Listeria; bacterial counts in spleen and liver were increased on days 3-5 by a factor of 10-1,000 in these organs. When examined histologically, organs from mice with the anti-TNF-alpha treatment contained more, and considerably bigger, lesions that exhibited central necrosis. The enhancing effect of anti-TNF-alpha on Listeria infection seemed greater early during Listeria infection on days 1-6 when compared to later phases of the infection around days 6-10. From the data presented we conclude that in addition to other lymphokines, such as IFN-gamma, TNF-alpha is of importance during the entire course of a Listeria infection in nu/nu mice.

To initiate an immune response after recognition of antigen presented in the context of class II antigens of the major histocompatibility complex (MHC) on macrophages, dendritic cells or B cells, T helper cells become activated. This process involves direct cell-cell contact mediated by adhesion reactions, eg, between the lymphocyte function-associated antigen-one (LFA-1) on T cells and its ligand, the intercellular adhesion molecule-1 (ICAM-l) on antigen presenter cells. This allows the triggering of the T cells by binding of antigen to the T cell antigen receptor/CD3 complex which transduces antigen-specific extracellular stimuli across the plasma membrane, generating intracellular signals. These events render T cells competent to receive progression signals to enter from G1 the S phase, a process mediated by interleukin-2 binding to its receptor. In addition to direct cell-cell contacts, soluble factors released by lymphocytes, monocytes-macrophages or by cells not belonging to the immune system (eg, keratinocytes or fibroblasts) are also involved in the maturation, growth and activation of the cellular elements of the immune system. Provided their synthesis by parenchymal cells of different organs, cytokines, polypeptide mediators that transmit signals from one cell to another, may propagate local expansion and activation of lymphocytes having infiltrated the tissue through the vessel wall. In the brain, T cell infiltrates can be observed mainly in viral diseases and multiple sclerosis.

Synthetic polypeptides corresponding to hydrophilic regions of human interferon gamma (HuIFN gamma) based on the amino acid sequence of HuIFN gamma inferred from its cDNA sequence were used to produce antibodies in rabbits which reacted with the polypeptides and which might also be expected to recognise native HuIFN gamma. Groups of 3 or 4 rabbits were immunised with synthetic polypeptides corresponding to HuIFN gamma amino acid sequences 1-20, 1-59, 24-59, 36-59 and 87-96 which included major hydrophilic domains of the IFN gamma molecule. All the rabbits produced antibodies which recognised the polypeptide immunogen, but to date only 1 of 4 rabbits immunised with polypeptide 24-59 and 1 of 3 rabbits immunised with polypeptide 1-59 have produced antibodies which also recognise native HuIFN gamma. The positively reacting antiserum from the rabbit immunised with polypeptide 24-59 could only be shown to weakly bind to HuIFN gamma, whereas the positively reacting antiserum from the rabbit immunised with polypeptide 1-59 was shown to both weakly bind to HuIFN gamma and weakly neutralise its in vitro antiviral effect. The results so far obtained suggest that the amino acid sequences close to the N-terminus are important for biological activity.

To evaluate the potential role of cachectin/TNF-alpha in the pathogenesis of bacterial and viral meningitis, concentrations and kinetics of TNF-alpha were determined in cerebrospinal fluid (CSF). After intracerebral, but not systemic, infection with Listeria monocytogenes in mice, TNF-alpha was detected as early as 3 h after infection reaching maximum titers after 24 h. However, TNF-alpha was not found in serum during the course of Listeria infection. In contrast to bacterial meningitis, no TNF-alpha was detected at any time in CSF of mice suffering from severe lymphocytic choriomeningitis induced by intracerebral infection with lymphocytic choriomeningitis virus. This difference is striking since both model infections led to a massive infiltration of polymorphonuclear and mononuclear leukocytes into the meninges and CSF. The results found for the two model infections were paralleled by findings in humans; CSF from three out of three patients with bacterial meningitis examined during the first day of hospitalization showed significant levels of TNF-alpha; none of the CSF obtained later than 3 d after hospitalization was positive. In addition, similarly to what was found in mice with viral meningitis, zero out of seven patients with viral meningitis had detectable TNF-alpha in CSF.

During generalized immune responses such as the acute phase of graft-vs.-host reaction (GvHR) or many systemic viral infections the macrophage-monocyte system of mice is activated as demonstrated by their increased bactericidal capacity against Listeria monocytogenes. To study the effect of interferon (IFN)-gamma in maintainance of macrophage activity, lymphocytic choriomeningitis virus (LCMV) primed C57BL/6 mice or (C57BL/10 x B10.BR) F1 mice undergoing a GvHR were treated with polyclonal specific sheep anti-IFN-gamma antiserum. In both cases injection of anti-IFN-gamma resulted in inhibition of the stimulatory effects on macrophages as detected by reduced bactericidal activity when compared to mice treated with normal serum. Treatment with a polyclonal sheep anti-IFN-alpha,beta antiserum on the other hand did not interfere with the activation status of the macrophages. The findings suggest that IFN-gamma is produced both early in a GvHR and during the acute phase of an systemic infection with LCMV, and that it is involved in in vivo modulation of the increased activity of mononuclear phagocytes in immunologically stimulated mice.

The lymphocytic choriomeningitis virus (LCMV) isolates Docile (D) and Aggressive (A) of Pfau et al. were studied in various strains of mice. Disease susceptibility, assessed as mortality and time to death to LCMV-D or -A varied greatly amongst mouse strains, and all four possible susceptibility patterns were observed: susceptibility to both (e.g. SWR/J), resistance to both (e.g. DBA/2), susceptibility to A but resistance to D (C57BL/6), or vice versa (CBA/J). Irrespective of the virus isolate or the mouse strain tested, susceptibility correlated with both early and high cytotoxic T cell activity found in spleens or leptomeningeal infiltrates, and with early and high primary footpad swelling reaction after local infection. C57BL/6 mice infected with A or SWR/J infected with A or with D showed, in both test systems, early and high activities; in contrast, DBA/2 mice infected with either D or A, and C57BL/6 infected with D showed no or only slow and low responses in both tests. Early and high LCMV-specific cytotoxic T cell activity, and the rapidity and extent of the primary footpad reaction directly correlated with susceptibility to LCM and all were dominantly regulated by H-2D.

Zunehmend beschaftigen sich Peptidchemiker mit der Synthese biologisch und medizinisch wichtiger Polypeptide. In diesem Aufsatz werden einige nutzliche Anwendungen von synthetischen Peptiden in Biochemie, Molekularbiologie, Immunologie und Medizin beschrieben. Synthetische Peptide sind von Interesse fur Untersuchungen uber Struktur-Funktions-Beziehungen bei Polypeptiden, als Peptidhormone und Hormonanaloga, fur die Herstellung kreuzreagierender Antikorper und fur die Konstruktion neuer Enzyme. Peptidchemiker konnen zum Fortschritt in den biologischen und medizinischen Wissenschaften beitragen, wenn sie mit deren aktuellen Problemen vertraut sind.

The influence of major transplantation antigens on susceptibility to T cell-mediated disease caused by infection with the noncytopathic virus lymphocytic choriomeningitis virus (LCMV) was evaluated in B10 H-2-congenic mice. Susceptibility to early T cell-mediated liver cell destruction (day 7-9) and early mortality (before day 12) was H-2Dq linked and correlated directly with early (day 6-8) and high cytotoxic T cell activity. In contrast, susceptibility to become an LCMV carrier, inability to rapidly clear virus, or tendency to develop late hepatitis (day 14-17) was linked to Dk and correlated with absence of early cytotoxic T cell activity. Thus, H-2D-regulated T cell-immune responses controlling both virus spread and immunopathology may directly determine the type and severity of disease. The results illustrate that susceptibility to disease caused by one virus may be linked to distinct MHC alleles dependent upon the disease parameter studied.

Although interferons (IFNs) were originally described as proteins conferring antiviral resistance to eukaryotic cells1, various additional biological effects have since been attributed to them, including inhibition of cell proliferation and modulation of the immune response2. More recently it was recognized that interferons from different sources have similar amino acid sequences, suggesting their evolutionary relationship. When the sequences of human interferons of the α and β type were aligned to give maximum homology, a largely conserved region was found near the carboxyl terminus of the molecules3,4. The homology of human leukocyte IFN-α1 and human fibroblast interferon between positions 111 and 166, for instance, was 40%. As conservation of segments of polypeptide chains during evolution may indicate their functional importance5, we synthesized three carboxyl-terminal fragments of human IFN-α1 (ref. 6) ranging in size from 33 to 96 amino acid residues. If the folding of these fragments was similar to that of the corresponding segments in the intact protein, it was conceivable that they had biological activity and that antibodies raised against the synthetic fragments cross-reacted with natural interferon. Such antibodies could be used for the affinity purification of human interferon produced in bacteria and for radioimmunoassays. As we show here, none of the fragments had antiviral activity. A mouse monoclonal antibody prepared against a 56-residue fragment also bound intact IFN-α1 and IFN-α2 without neutralizing their antiviral effect. Thus, this antibody, directed against a largely conserved region, may show substantial cross-reactivity within the interferon protein family.

Synthesis of B cell-stimulating factor-2 (BSF-2) and IFN-gamma was shown in cerebrospinal fluids (CSF) collected from mice with experimental viral meningitis. In the CSF, the level of BSF-2 started to increase 24 h after intracerebral infection with lymphocytic choriomeningitis virus (LCMV) with rapid increase after day 4. IFN-gamma was not detected in the CSF before day 5 or 6 after infection, but increased sharply thereafter. In athymic nude mice, LCMV infection did not result in meningitis, and both BSF-2 and IFN-gamma levels were only slightly and transiently elevated. These findings suggest that activated mature T cells are required for development of disease and production of both BSF-2 and IFN-gamma. As observed in mice, BSF-2 was also detected in 16 out of 19 CSF samples collected from patients with acute viral infections of the central nervous system (CNS). Intrathecal production of BSF-2 and IFN-gamma may be instrumental in local production of antiviral antibodies by B lymphocytes/plasma cells invading the CNS during viral CNS disease.

A model for immunologically T cell-mediated hepatitis was established in mice infected with lymphocytic choriomeningitis virus (LCMV). The severity of hepatitis was monitored histologically and by determination of changes in serum levels of the enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH), and alkaline phosphatase (AP). Kinetics of histological disease manifestations, increases of liver enzyme levels in the serum, and cytotoxic T cell activities in livers and spleens all correlated and were dependent upon several parameters: LCMV-isolate; LCMV-WE caused extensive hepatitis, LCMV-Armstrong virtually none. Virus dose. Route of infection; i.v. or i.p. infection caused hepatitis, whereas infection into the footpad did not. The general genetic background of the murine host; of the strains tested, Swiss mice and A-strain mice were more susceptible than C57BL or CBA mice; BALB/c and DBA/2 mice were least susceptible. The degree of immunocompetence of the murine host; T cell deficient nu/nu mice never developed hepatitis, whereas nu/+ or +/+ mice always did. B cell-depleted anti-IgM-treated mice developed immune-mediated hepatitis comparably or even more extensively than control mice. Local cytotoxic T cell activity; mononuclear cells isolated from livers during the period of overt hepatitis were two to five times more active than equal numbers of spleen cells. Adoptive transfer of nylon wool-nonadherent anti-Thy-1.2 and anti-Lyt-2 plus C-sensitive, anti-L3T4 plus C-resistant lymphocytes into irradiated mice preinfected with LCMV-WE caused a rapid time- and dose-dependent linear increase of serum enzyme levels. This increase was caused by adoptive transfer of lymphocytes if immune cell donors and recipient mice shared class I, but not when they shared class II histocompatibility antigens. The donor cell dose-dependent increase of these enzymes was first measurable 6-18 h after transfer with 2 X 10(8) cells or 3 X 10(6) cells, respectively. The time-dependent increase caused by the adoptive transfer of 1-2 X 10(8) cells was strictly linear during a period of up to 25-40 h. These results indicate single-hit kinetics of liver cell death and suggest that effector T cells destroy infected liver cells via direct contact rather than via soluble toxic mediators. The results may represent the best in vivo correlate of the in vitro 51Cr-release assay that has been analyzed so far, and strongly support the view that antiviral cytotoxic T cells are directly cytolytic in vivo.(ABSTRACT TRUNCATED AT 400 WORDS)

Interleukin 6 (IL 6) was found to be produced in the central nervous system (CNS) of ICR +/+ mice infected with lymphocytic choriomeningitis virus (LCMV) or with vesicular stomatitis virus (VSV). When infecting athymic ICR nu/nu mice which cannot develop T cell-mediated meningitis after LCMV infection, no significant synthesis of IL 6 was detected in the CNS. IL 6 was found, however, to be produced intrathecally in ICR nu/nu mice infected with VSV, which causes a T cell-independent acute encephalitis. This suggested that IL 6 may also originate from cells not belonging to the T cell compartment. Indeed, in vitro assays showed that both virus-infected microglial cells and astrocytes secreted IL 6. In astrocytes, the infection resulted in the induction of the 1.3-kb messenger RNA IL 6. Besides its effect on the development of B cell immunity in the brain, IL 6 may be involved in repair mechanisms initiated in the course of viral-induced tissue damage. As shown here, IL 6 induced an increase of the secretion of a neurotrophic factor, nerve growth factor by astrocytes. Thus, the intrathecal synthesis of IL 6 may be part of the host response to infection favoring immune-mediated elimination of the infectious agent as well as trophic support for neurons.

Four oligopeptides corresponding to overlapping regions of the human leucocyte interferon alpha 1 (IFN) amino acid sequence have been synthesized by the solid-phase method. The physicochemical properties of these peptides were investigated by analytical ultracentrifugation and circular dichroism, absorbance, and fluorescence spectroscopies. IFNs 24-81 and 111-166 were shown to possess considerable ordered structure in solution. IFN 111-166 appeared to adopt a beta-stranded conformation, which could be reversibly unfolded by the addition of urea. Equimolar mixtures of IFNs 24-81 and 111-166 formed a noncovalent complex that was resistant to the formation of the Cys29-Cys139 disulfide bond found in native interferon alpha 1. Predictions of the secondary structure of interferon alpha 1 were made and the results compared to the measured properties of the fragments. In general, it was found that the prediction methods favored a highly alpha-helical conformation.