Remitting Multiple Sclerosis Research Articles

Circulatory Indicators of Lipid Peroxidation, the Driver of Ferroptosis, Reflect Differences between Relapsing–Remitting and Progressive Multiple Sclerosis

Ferroptosis, a lipid peroxidation- and iron-mediated type of regulated cell death, relates to both neuroinflammation, which is common in relapsing-remitting multiple sclerosis (RRMS), and neurodegeneration, which is prevalent in progressive (P)MS. Currently, findings related to the molecular markers proposed in this paper in patients are scarce. We analyzed circulatory molecular indicators of the main ferroptosis-related processes, comprising lipid peroxidation (malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), and hexanoyl–lysine adduct (HEL)), glutathione-related antioxidant defense (total glutathione (reduced (GSH) and oxidized (GSSG)) and glutathione peroxidase 4 (GPX4)), and iron metabolism (iron, transferrin and ferritin) to estimate their contributions to the clinical manifestation of MS and differences between RRMS and PMS disease course. In 153 patients with RRMS and 69 with PMS, plasma/serum lipid peroxidation indicators and glutathione were quantified using ELISA and colorimetric reactions, respectively. Iron serum concentrations were determined using spectrophotometry, and transferrin and ferritin were determined using immunoturbidimetry. Compared to those with RRMS, patients with PMS had decreased 4-HNE (median, 1368.42 vs. 1580.17 pg/mL; p = 0.03). Interactive effects of MS course (RRMS/PMS) and disease-modifying therapy status on MDA (p = 0.009) and HEL (p = 0.02) levels were detected. In addition, the interaction of disease course and self-reported fatigue revealed significant impacts on 4-HNE levels (p = 0.01) and the GSH/GSSG ratio (p = 0.04). The results also show an association of MS course (p = 0.03) and EDSS (p = 0.04) with GSH levels. No significant changes were observed in the serum concentrations of iron metabolism indicators between the two patient groups (p > 0.05). We suggest circulatory 4-HNE as an important parameter related to differences between RRMS and PMS. Significant interactions of MS course and other clinically relevant parameters with changes in redox processes associated with ferroptosis support the further investigation of MS with a larger sample while taking into account both circulatory and central nervous system estimation.

Different genome-wide DNA methylation patterns in CD4+ T lymphocytes and CD14+ monocytes characterize relapse and remission of multiple sclerosis: Focus on GNAS

BackgroundRelapsing-remitting multiple sclerosis (RRMS) is a most common form of multiple sclerosis in which periods of neurological worsening are followed by periods of clinical remission. RRMS relapses are caused by an acute autoimmune inflammatory process, which can occur in any area of the central nervous system. Although development of exacerbation cannot yet be accurately predicted, various external factors are known to affect its risk. These factors may trigger the pathological process through epigenetic mechanisms of gene expression regulation, first of all, through changes in DNA methylation. MethodsIn the present work, we for the first time analyzed genome-wide DNA methylation patterns in CD4+ T lymphocytes and CD14+ monocytes of the same RRMS patients in relapse and remission. The effects of the differential methylation on gene expression were studied using qPCR. ResultsWe found 743 differentially methylated CpG positions (DMPs) in CD4+ cells and only 113 DMPs in CD14+ cells. They were mostly hypermethylated in RRMS relapse in both cell populations. However, the proportion of hypermethylated DMPs (as well as DMPs located within or in close proximity to CpG islands) was significantly higher in CD4+ T lymphocytes. In CD4+ and CD14+ cells we identified 469 and 67 DMP-containing genes, respectively; 25 of them were common for two cell populations. When we conducted a search for differentially methylated genomic regions (DMRs), we found a CD4+ specific DMR hypermethylated in RRMS relapse (adj. p = 0.03) within the imprinted GNAS locus. Total level of the protein-coding GNAS transcripts in CD4+ T cells decreased significantly in the row from healthy control to RRMS remission and then to RRMS relapse (adj. p = 3.1 × 10–7 and 0.011, respectively). ConclusionOur findings suggest that the epigenetic mechanism of DNA methylation in immune cells contributes to the development of RRMS relapse. Further studies are now required to validate these results and shed light on the molecular mechanisms underlying the observed GNAS methylation and expression changes.

SENSITIVITY OF PLATELET-LYMPHOCYTE RATIO, MONOCYTE-LYMPHOCYTE RATIO, NEUTROPHIL-LYMPHOCYTE RATIO, RED BLOOD CELL DISTRIBUTION WIDTH AND SYSTEMIC IMMUNE INFLAMMATORY INDEX IN MULTIPLE SCLEROSIS PATIENTS: RELATIONSHIP WITH DISEASE SUBTYPES AND DISABILITY

OBJECTIVE: Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system (CNS). Although it is not clear how autoimmune responses occur, it is known that peripheral blood cells may promote the initiation and exacerbation of the inflammatory response in MS. The aim of this study was to evaluate the sensitivity and specificity of neutrophil-lymphocyte ratio (NLR), monocyte-lymphocyte ratio (MLR), platelet-lymphocyte ratio (PLR), erythrocyte distribution width (RDW) and systemic immune inflammation index (SIII) in MS and to determine their relationship with disability. MATERIAL AND METHODS: In this single-center, retrospective case-control study, 104 patients with MS and 65 healthy individuals were included. Disability was assessed with the Expanded Disability Status Scale (EDSS) score, and patients were classified as mild (EDSS<3), moderate (3.5 ≤ EDSS ≥4.5), and severe (EDSS≥5) disabilities. NLR, MLR, PLR, RDW and SIII values were calculated according to the complete blood counts of the patients and the control group. Inflammation markers between the control group and MS patients, MS subtypes, and relapsing-remitting MS (RRMS) attack and remission periods were compared. the relationship between these markers and duration of illness, number of attacks and disability was examined. RESULTS: MLR and RDW were higher in MS than the controls. There was no significant difference in these values between MS subtypes. It was observed that RDW increased as the disease duration, number of attacks and EDSS increased. CONCLUSIONS: In our study, in evaluating inflammation it was seen that MLR and RDW were more determinative than PLR, NLR and SIII. The fact that RDW is higher in MS patients and is affected by disease disability indicates that RDW may have an important role in the follow-up of MS patients and in evaluating the degree of disability.

Regional hippocampal atrophy reflects memory impairment in patients with early relapsing remitting multiple sclerosis

BackgroundResearch work has shown that hippocampal subfields are atrophic to varying extents in multiple sclerosis (MS) patients. However, studies examining the functional implications of subfield-specific hippocampal damage in early MS are limited. We aim to gain insights into the relationship between hippocampal atrophy and memory function by investigating the correlation between global and regional hippocampal atrophy and memory performance in early MS patients.MethodsFrom the Italian Neuroimaging Network Initiative (INNI) dataset, we selected 3D-T1-weighted brain MRIs of 219 early relapsing remitting (RR)MS and 246 healthy controls (HC) to identify hippocampal atrophic areas. At the time of MRI, patients underwent Selective-Reminding-Test (SRT) and Spatial-Recall-Test (SPART) and were classified as mildly (MMI-MS: n.110) or severely (SMI-MS: n:109) memory impaired, according to recently proposed cognitive phenotypes.ResultsEarly RRMS showed lower hippocampal volumes compared to HC (p < 0.001), while these did not differ between MMI-MS and SMI-MS. In MMI-MS, lower hippocampal volumes correlated with worse memory tests (r = 0.23–0.37, p ≤ 0.01). Atrophic voxels were diffuse in the hippocampus but more prevalent in cornu ammonis (CA, 79%) than in tail (21%). In MMI-MS, decreased subfield volumes correlated with decreases in memory, particularly in the right CA1 (SRT-recall: r = 0.38; SPART: r = 0.34, p < 0.01). No correlations were found in the SMI-MS group.ConclusionHippocampal atrophy spreads from CA to tail from early disease stages. Subfield hippocampal atrophy is associated with memory impairment in MMI-MS, while this correlation is lost in SMI-MS. This plays in favor of a limited capacity for an adaptive functional reorganization of the hippocampi in MS patients.

Clinical and Immunological Impact of Ocrelizumab Extended Interval Dosing in Multiple Sclerosis: A Single-Center, Real-World Experience.

Ocrelizumab (OCR), an anti-CD20 monoclonal antibody, is approved for treating relapsing remitting (RR) and primary progressive (PP) multiple sclerosis (MS). The standard interval dosing (SID) regimen requires intravenous infusions every six months. Experience of extended dosing due to COVID-19 pandemic-related issues suggests that this strategy may provide comparable efficacy while reducing treatment burden and healthcare costs. This study aimed to evaluate clinical effectiveness, changes in B- and T-cell count, and immunoglobulin dynamics associated with extended interval dosing (EID) of ocrelizumab in a real-world setting. We retrospectively included RRMS or PPMS patients treated with OCR that had already received two OCR cycles and with at least 6 months of follow up after the last infusion. EID was defined as a ≥4 weeks delay compared to SID. Clinical outcomes were occurrence of relapses, MRI activity, 6-months confirmed disability progression (CDP) and their combination (No Evidence of Disease Activity, NEDA-3). We also evaluated changes in CD19+ B cell count, CD4+ and CD8+ T cell count, immunoglobulin titers, and occurrence of hypogammaglobulinemia (hypo-Ig). Frequency tests, multivariate regression models, and survival analysis were applied as appropriate. We analyzed data on 93 subjects (75.3% RRMS) for a total of 389 infusions (272 SID, 117 EID). Clinical and MRI activity, CDP, and NEDA 3 did not significantly differ between EID and SID. EID was associated with lower rates of B-cell depletion. T-cell dynamics and incidence of hypo-Ig were comparable following EID and SID. Hypo-IgG at index infusion was associated with further occurrence of hypo-IgG; male sex and hypo-IgM at index infusion were independently associated with hypo-IgM. In conclusion, OCR EID does not impact MS clinical and radiological outcomes, although it interferes with B-cell dynamics. These findings provide support for a tailored schedule of OCR in MS.

Real-world Data on Ocrelizumab Treatment: Efficacy, Safety, and Tolerability Analysis in Patients with Relapsing Remitting and Primary Progressive Multiple Sclerosis in Southeastern Ontario (P2-6.014)

Real-world Data on Ocrelizumab Treatment: Efficacy, Safety, and Tolerability Analysis in Patients with Relapsing Remitting and Primary Progressive Multiple Sclerosis in Southeastern Ontario (P2-6.014)

Cryptococcal Cerebellitis in the Setting of Fingolimod Use for Multiple Sclerosis: Atypical Presentation of Dystonia and Imaging Finding Misinterpreted as Subacute Ischemic Stroke

Objective: Describe a case of cryptococcal cerebellitis in a patient on Fingolimod with atypical findings of generalized dystonia and imaging findings mistaken for bilateral subacute cerebellar strokes. Background: Fingolimod is used for multiple sclerosis (MS) and is often associated with lymphopenia with prolonged use. Cryptococcal opportunistic infections in patients on Fingolimod, have been rarely described. Design/Methods: Case report and review of literature. Results: 44-year-old man with MS in remission treated with Fingolimod for the last 8 years, and poorly controlled type-II diabetes mellitus (A1c 10.4), transferred to our facility for stroke work-up with brain MRI findings reportedly showing subacute bicerebellar ischemic strokes. On exam, he was somnolent and intermittently followed commands. He had nuchal rigidity and positive Kernig and Brudzinski signs. Meningitis was suspected and empiric treatment was initiated. MRI findings on DWI/ADC/FLAIR sequences were inconsistent with any vascular territory. Leptomeningeal enhancement was noted in the cerebellar folia, basal cisterns, suprasellar cistern, and brainstem surface. Labs were remarkable for lymphopenia (250), cerebrospinal fluid (CSF) and blood culture grew cryptococcus neoformans, so treatment with liposomal amphotericin B and flucytosine was started. On day 4, patient developed generalized dystonia that resolved with diphenhydramine. Unfortunately, patient did not respond to therapy as evident on repeat LP on day 6. His physical exam worsened, and he was intubated 2 days after. Repeat MRI showed with tonsillar herniation and patient expired. Conclusion: Cryptococcal cerebellitis is a rare opportunistic infection occurring in MS patients on Fingolimod. Early recognition is crucial to avoid rapidly worsening neurological outcomes.

Quality of life of patients with multiple sclerosis after high-dose immunosuppressive therapy with autologous hematopoietic stem cell transplantation

Introduction. Multiple sclerosis (MS) is a chronic autoimmune and neurodegenerative disease of the central nervous system (CNS). High-dose immunosuppressive therapy with autologous hematopoietic stem cell transplantation (HDIT–AHSCT) is the most eff ective method of treating MS, which has a positive eff ect on the quality of life of patients and inhibition of the progression of disability.The aim of the study was to analyze the dynamics of quality of life in patients with MS who underwent the HDIT– AHSCT method in groups with diff erent clinical effi cacy of this method.Material and methods. The single-center observational study included patients with MS (n = 21, mean age 35.5 ± 6.2 (28–50) years; men/women — 11/10; remitting MS (RMS) n = 16, secondary-progressive (SPMS) n = 4, primaryprogressive (PPMS) n = 1), after HDIT–AHSCT (cyclophosphamide-rituximab conditioning regimen). Assessment of the quality of life (SF-36, FAMS, CSP-MS42, HADS) and the degree of disability in all patients was carried out before and 12 months after HDIT–AHSCT.Results. 12 months after HDIT–AHSCT, more than 50% of patients showed signifi cant improvement in quality of life according to the scales of viability and social functioning of the SF-36 questionnaire, as well as according to the scales of general well-being and additional problems of the FAMS questionnaire. 57% of patients had a signifi cant improvement in the overall FAMS score. According to the CSP-MS42 questionnaire, a decrease in weakness (p &lt; 0.05), feelings of constant fatigue and rapid fatigue during exercise was revealed. After transplantation, there was a slight decrease in anxiety (p = 0.91) and a signifi cant decrease in depression (p = 0.011).Conclusion. The expected dynamics was established: in patients with clinical improvement, there was an improvement in the quality of life for most indicators; in patients with stabilization, most quality of life indicators were similar before and after HDIT–AHSCT; with clinical deterioration after HDIT–AHSCT, quality of life parameters worsened. These data indicate the expediency of further optimization of patient selection criteria and various stages of the HDIT–AHSCT in order to obtain optimal clinical results and, accordingly, improve the quality of life of patients.

Hypogammaglobulinemia and Infection Risk in an Ocrelizumab-treated Multiple Sclerosis Cohort.

Ocrelizumab is an effective anti-CD20 therapy approved for Relapsing Remitting (RRMS) and Primary Progressive Multiple Sclerosis (PPMS). In clinical trials, a proportion of patients developed hypogammaglobulinemia which could contribute to infection risk. This study aimed to identify hypogammaglobulinemia and its risk factors and evaluate potentially associated serious infection risk in a real-world cohort of patients. All MS patients treated with ocrelizumab in a Quebec City MS clinic from January 2017 to August 2021 were included and detailed patient characteristics were collected by chart review. Levels of immunoglobulins (IgM, IgA and IgG) were assessed prior to each treatment. Serious infection was defined as an infection requiring hospitalization or emergency room treatment. Association between hypogammaglobulinemia and serious infection was analyzed. A total of 266 patients (average follow-up 2.05 years) were included (87% RRMS). After 6 infusions, 32.8%, 3.5% and 4.2% of patients had at least one IgM, IgA and IgG hypogammaglobulinemia event respectively. Aside from pre-treatment hypogammaglobulinemia, there were no variables associated with on-treatment hypogammaglobulinemia. There was a total of 21 serious infections (3.36 and 12.33 per 100-person-years in RRMS and PPMS). Developing hypogammaglobulinemia during treatment was not associated with serious infection. A regression analysis did not show associations between serious infection and key disease characteristics. Similar to ocrelizumab extension studies, our cohort demonstrated a significant rate of hypogammaglobulinemia over time, mostly with IgM. No association was found between hypogammaglobulinemia and serious infection.

Levels of circulating cytokines in children with multiple sclerosis with different effectiveness of interferon therapy

Multiple sclerosis (MS) is a chronic, demyelinating disease that leads to disability. Understanding the etiology of MS contributes to the development of pathogenetic methods of treatment, and the search for informative biomarkers of the effectiveness of treatment will allow the patient to adjust therapy in time. The aim of this work was to determine informative cytokines and cytokine profiles to predict the effectiveness of IFN-β1a therapy in children with MS.Materials and methods. 66 children with MS aged 16 [14.2–17.5] years who are on INFß-1a therapy were examined: group 1 — patients with exacerbation of MS (with active foci of demyelination by MRI), n = 34; group 2 — patients in remission of MS (without active foci), n = 32. The content of cytokines in the blood serum of patients was assessed using the multiplex panel Human Th17 Magnetic Bead Panel.Results: There was a significant increase in cytokine concentrations in patients with exacerbation of MS compared with children in remission: IL5, IL6, IL9, IL12p70, IL17E/IL25, IL21, IL28A, GM-CSF, TNFß. Threshold values for IL9 (AUC = 0,785), IL6 (AUC = 0,750), TNFß (0,740), IL28A (AUC = 0,744) were obtained above which it is possible to predict an exacerbation of MS in patients: IL9 — 3.9 pg/ml (Sn — 70.6, Sp — 71.9), IL6 — 4.0 pg/ml (Sn — 70.6, Sp — 68.8), TNFß — 6.6 pg/ml (Sn — 70.6, Sp — 71.9), IL28A — 243.0 pg/ml (Sn — 70.6, Sp — 71.9). Cytokine profiles associated with T-lymphocytes and their functions were evaluated using z-score.Conclusions. For the first time, an increase in cytokine levels was demonstrated in children with active foci of demyelination compared to patients in remission of MS. An increase in proinflammatory cytokines and cytokine profiles associated with Th1 and Th17, as well as with Th2 and Th22 has been shown. The use of threshold values for IL9, IL6, TNFβ, IL28A will help predict the development of exacerbation in patients with MS.

Glial fibrillary acidic protein: is there a relationship with the course of multiple sclerosis and traumatic brain injury before the onset of multiple sclerosis?

To evaluate the relationship between the level of glial fibrillary acidic protein (GFAP) in the blood and the rate of progression (RP) of neurological disorders and exacerbation frequency of multiple sclerosis (MS) in the presence/absence of traumatic brain injury (TBI) before the onset of MS. Caucasians born and living in the Altai region of Russia with relapsing-remitting MS in remission took part in a cross-sectional observational randomized study: 43 patients without a history of TBI and 43 patients with TBI before the onset of MS (mean age 36.1±9.3 and 33.5±8.2 years, MS duration 7.6±6.6 and 5.9±4.6 years, respectively). Intracranial injury occurred 14.8±7.8 years before the onset of MS and was documented as a concussion in 76% of patients. The patient groups did not differ in the frequency of MS exacerbations, while RP was higher in the group with a history of TBI (p=0.013). There were no intergroup differences in GFAP levels, as well as no correlations between GFAP levels and RP (p=0.520 and p=0.255 in groups with/without TBI, respectively) and the exacerbation frequency. No differences were found between subgroups of patients with different clinical forms of TBI in RP and GFAP levels (H=0.26; p=0.880 and H=1.48; p=0.476, respectively). There was no correlation of the TBI duration with RP (p=0.902) and GFAP level (p=0.413). The results indicate a low probability of a connection between the level of GFAP in the blood of patients with MS during the period of remission, both with the possible long-term consequences of a TBI occurred before the onset of MS, and during the course of MS.

Content of myeloid-derived suppressor cells in autoimmune diseases in children

Myeloid-derived suppressor cells (MDSCs) play an important role in regulation of immune response. An increase in their number in adult patients with autoimmune diseases has been reported. G-MDSCs, M-MDSCs, and MDSCs(M-G-) at different stages of autoimmune disease may both activate T cell proliferation, leading to disease progression, or inhibit it, thus promoting Treg differentiation. Arginase-1 (Arg- 1) is an enzyme in MDSCs that reduces the concentration of arginine required for T lymphocyte proliferation. Our aim was to evaluate the content of MDSCs populations and functional activity of MDSCs in children with autoimmune diseases. 75 children with inflammatory bowel diseases (IBD), 60 children with multiple sclerosis (MS), 69 children with psoriasis (PS), 62 healthy age-matched children were included into the study. The content of MDSCs ((CD3, CD19, CD56, HLA-DR)-, CD11b+ and CD33+), subpopulations of MDSCs (M-MDSCs, G-MDSCs expressing CD14 and CD15), assessment of Arg-1 activity were performed by flow cytometry techniques. The content of MDSCs in patients with IBD, MS and PS was significantly higher than in the comparison group and depended on the state of exacerbation/remission. In exacerbation and remission of IBD, MS and PS, a significant increase of MDSCs was revealed when compared with healthy children; the highest values were found in children in exacerbation of MS (Me-3.5 (2.5-5.6) % MNC against Me-1.6 (0.9-2.5) % MNC, p 0.001). In patients with MS, the content of G-MDSC, M-MDSC was significantly higher, and MDSC(M-G-) was lower than in healthy children. An increase in absolute amounts of G-MDSCs was shown in MS exacerbation compared to the disease remission state (p = 0.022). For patients with IBD, a significant increase in percentage of MDSCs and M-MDSCs (p = 0.014 and p = 0.045, respectively) was obtained in exacerbation of the disease relative to remission state. In patients with IBD, MS, and PS, a significant increase in Arg-1 activity in MDSCs was found, with a decreased number of MDSCs in patients in remission compared to exacerbation phase of the disease. In children with autoimmune diseases, an increase in the MDSC populations was found. The activity of arginase-1 in MDSCs is increased in remission, along with a decrease in their numbers.

Continued dysregulation of the B cell lineage promotes multiple sclerosis activity despite disease modifying therapies

A clear understanding of the origin and role of the different subtypes of the B cell lineage involved in the activity or remission of multiple sclerosis (MS) is important for the treatment and follow-up of patients living with this disease. B cells, however, are dynamic and can play an anti-inflammatory or pro-inflammatory role, depending on their milieu. Depletion of B cells has been effective in controlling the progression of MS, but it can have adverse side effects. A better understanding of the role of the B cell subtypes, through the use of surface biomarkers of cellular activity with special attention to the function of memory and other regulatory B cells (Bregs), will be necessary in order to offer specific treatments without inducing undesirable effects.

INVESTIGATION OF THE EFFECTS OF CX3CL1 ON IMMUNOPATHOGENESIS AND COGNITIVE FUNCTIONS IN RELAPSING REMITTINGAND SECONDARY PROGRESSIVE MULTIPLE SCLEROSIS PATIENTS IN TERMS OF THE POSSIBILITY OF A BIOMARKER

Objectives. CX3CL1 is a unique chemokine with direct relationship with its receptor to reduce expression of proinflammatory genes inactivated microglia.It is constitutively expressed by healty neurons and defined as neuroimmune regulatory protein.CX3CL1/CX3CR1signaling has been linked to human neurodegeneration. Under physiological conditions, disruption of this signaling leads to impairments inmotor learning, cognitive function, and synaptic plasticity. CX3CR1 also plays a role in the regulation of hippocampal-dependent memoryformation.Our aim in this study is to evaluate the role of CX3CL1 on immunopathogenesis and cognitive functions in Relapsing Remitting (RRMS) and Secondary Progressive Multiple Sclerosis (SPMS) patients in terms of biomarker possibility. Methods. Eight RRMS, 24 SPMS and 30 healty controls have been participated in this study. RRMS and SPMS were diagnosed according toMc Donald 2017 criteria. The degree of neurological deficits were assessed by EDSS.Serum levels of CX3CL1 were measured by ELISAmethod.The severity of cognitive impairment in patients were evaluate with MoCA test.Statistical analysis were performed using the IBMSPSS21. Results. When the RRMS, SPMS and control groups were compared,it was observed that the serum CX3CL1 levels in the SPMS groupwere decreased statistically significant.In addition,there was a positive correlation between MoCA score and serum CX3CL1 level in thepatients with RRMS and SPMS. Conclusion. Cytokines and chemokines play an important role in the immunopathogenesis of multiple sclerosis.Amongchemokines, serum levels of CX3CL1 was reported to be elevated in RRMS patients. However, it has not been observed in patients SPMS. CX3CL1 is an intrinsic neuroprotective chemokine and inhibitor against neurotoxicity. According to the results of our study, thedecrease in serum CX3CL1 levels serum CX3CL1 supports the development of neurodegeneration related neuroinflammation. In addition, thepositive correlation of MoCA test with CX3CL1 levels suggests that it has an important role in cognition. As a result, our study shows that CX3CL1 can be used as an early biomarker associated with disease progression and cognition, additionally it will be promising newtherapeutic target.

Pain and participation in social activities in people with relapsing remitting and progressive multiple sclerosis.

Differences in pain between subtypes of multiple sclerosis are understudied. To compare the prevalence of pain, and the association between pain and: (a) pain interference and (b) social participation in people with relapsing-remitting multiple sclerosis and progressive multiple sclerosis. Participants completed the McGill Pain Questionnaire Short-Form-2, Pain Effects Scale and Ability to Participate in Social Roles and Activities-V2.0 questionnaires. We tested the association between multiple sclerosis subtype, pain severity, and pain interference/social participation using quantile regression. Of 231 participants (relapsing-remitting multiple sclerosis: 161, progressive multiple sclerosis: 70), 82.3% were women. The prevalence of pain was 95.2%, of more than mild pain was 38.1%, and of pain-related limitations was 87%; there were no differences between multiple sclerosis subtypes. Compared to participants with relapsing-remitting multiple sclerosis, those with progressive multiple sclerosis reported higher pain interference (mean (standard deviation) Pain Effects Scale; progressive multiple sclerosis: 15[6.0] vs relapsing-remitting multiple sclerosis: 13[5], p = 0.039) and lower social participation (Ability to Participate in Social Roles and Activities T-scores 45[9.0] vs 48.3[8.9], p = 0.011). However, on multivariable analysis accounting for age, physical disability, mood/anxiety and fatigue, multiple sclerosis subtype was not associated with differences in pain interference or social participation. Pain was nearly ubiquitous. Over one-third of individuals with relapsing-remitting multiple sclerosis and progressive multiple sclerosis reported pronounced pain, although this did not differ by multiple sclerosis subtype.

Involvement of Degenerating 21.5 kDa Isoform of Myelin Basic Protein in the Pathogenesis of the Relapse in Murine Relapsing-Remitting Experimental Autoimmune Encephalomyelitis and MS Autopsied Brain.

Multiple sclerosis (MS) is the chronic inflammatory demyelinating disease of the CNS. Relapsing-remitting MS (RRMS) is the most common type of MS. However, the mechanisms of relapse and remission in MS have not been fully understood. While SJL mice immunized with proteolipid protein (PLP) develop relapsing-remitting experimental autoimmune encephalomyelitis (RR-EAE), we have recently observed that some of these mice were resistant to the active induction of relapsing EAE after initial clinical and histological symptoms of EAE with a severity similar to the relapsing EAE mice. To clarify the mechanism of relapsing, we examined myelin morphology during PLP139-151-induced RR-EAE in the SJL mice. While RR-EAE mice showed an increased EAE severity (relapse) with CNS inflammation, demyelination with abnormal myelin morphology in the spinal cord, the resistant mice exhibited a milder EAE phenotype with diminished relapse. Compared with the RR-EAE mice, the resistant mice showed less CNS inflammation, demyelination, and abnormalities of the myelin structure. In addition, scanning electron microscopic (SEM) analysis with the osmium-maceration method displayed ultrastructural abnormalities of the myelin structure in the white matter of the RR-EAE spinal cord, but not in that of the resistant mice. While the intensity of myelin staining was reduced in the relapsing EAE spinal cord, immunohistochemistry and immunoblot analysis revealed that the 21.5 kDa isoform of degenerating myelin basic protein (MBP) was specifically induced in the relapsing EAE spinal cord. Taken together, the neuroinflammation-induced degenerating 21 kDa isoform of MBP sheds light on the development of abnormal myelin on the relapse of MS pathogenesis.

Cryptococcal Cerebellitis In the Setting of Fingolimod Use for Multiple Sclerosis: Atypical Clinical Presentation of Dystonia and Imaging Finding Mimicking Subacute Ischemic Stroke (P1-10.004)

<h3>Objective:</h3> To describe a case of cryptococcal cerebellitis in a patient on Fingolimod with atypical findings of generalized dystonia and imaging findings mistaken for bilateral subacute cerebellar strokes. <h3>Background:</h3> Fingolimod is used for multiple sclerosis and is often associated with lymphopenia with prolonged use. Opportunistic infections, particularly cryptococcus neoformans in patients on Fingolimod, have been rarely described. <h3>Design/Methods:</h3> Case report and review of literature. <h3>Results:</h3> 44-year-old man with multiple sclerosis in remission treated with Fingolimod since 2014, and poorly controlled type-II diabetes mellitus (A1c 10.4), transferred to our facility for stroke work-up with brain MRI findings reportedly consistent with subacute bilateral cerebellar ischemic strokes. On physical exam, he was somnolent and intermittently followed simple commands. He also had nuchal rigidity and positive Kernig and Brudzinski signs. Meningitis was suspected and empiric treatment was initiated. MRI findings on DWI/ADC/FLAIR sequences were inconsistent with any vascular territory. Additionally, extensive leptomeningeal enhancement was noted in the cerebellar folia, basal cisterns, suprasellar cistern, and the surface of the brainstem. Labs were remarkable for lymphopenia (Lymphocyte count 250), and both cerebral spinal fluid (CSF) and blood culture grew cryptococcus neoformans so treatment with liposomal amphotericin B and flucytosine was started. On day 4 of admission, patient developed generalized dystonia that resolved with diphenhydramine. Unfortunately, patient did not respond to therapy as evident on repeat LP on day 6 of admission. His physical exam continued to worsen, and he was intubated 2 days after. Repeat MRI was consistent with tonsillar herniation and patient expired <h3>Conclusions:</h3> Cryptococcal cerebellitis is a rare opportunistic infection occurring in multiple sclerosis patients on Fingolimod. Early recognition is crucial given rapidly worsening neurological outcomes and atypical presentation of generalized dystonia. <b>Disclosure:</b> Dr. Singh has nothing to disclose. Dr. Ahmed has nothing to disclose. Dr. Kakawand has nothing to disclose. Dr. Asif has nothing to disclose. Dr. Al-Awwad has nothing to disclose.

A comparison of DNA methylation profiles of blood mononuclear cells in patients with multiple sclerosis in remission and relapse

To study the whole-genome DNA methylation profiles of peripheral blood mononuclear blood cells (PBMCs) of patients with relapsing-remitting multiple sclerosis (RRMS) in remission and relapse in order to assess the contribution of this epigenetic mechanism of gene expression regulation to the activity of the pathological process. Eight patients with RRMS in remission and 6 patients in relapse were included in the study. Methylation levels of DNA CpG sites in PBMCs were analyzed using Infinium HumanMethylation450 BeadChip DNA microarrays. Seven differentially methylated positions (DMPs) were identified, of which 3 were hypermethylated (cg02981003, cg18486102, cg19533582) and 4 were hypomethylated (cg16814680, cg1964802, cg18584440, cg08291996) during RRMS relapse. Five DMPs are located in protein-coding genes (GPR123, FAIM2, BTNL2, ZNF8, ASAP2), one in microRNA gene (MIR548N), and one in an intergenic region. For all identified DMPs, we observed a change in DNA methylation levels of more than 20% (range 20.2-57.5%). Hierarchical clustering of DNA samples on the heatmap shows their clear aggregation into separate clusters corresponding to RRMS patients in the stages of relapse and remission. For the first time it was shown that during relapse and remission of RRMS there are differences in the DNA methylation profile that allow discrimination between these clinical stages. These data indicate the involvement of the epigenetic mechanism of DNA methylation in the activation of the pathological process in RRMS.

Microbiota markers level in the blood and cerebrospinal fluid of patients with different types of multiple sclerosis and radiologically isolated syndrome

To assess the level of microbiota markers in the blood and cerebrospinal fluid (CSF) of patients with different types of multiple sclerosis (MS), people with radiologically isolated syndrome (RIS) and control subjects. We used gas chromatography-mass spectrometry (GC-MS) to evaluate the levels of microbiota markers in 69 patients with different types of MS (27 patients in the acute stage, 35 patients with MS in remission, 7 patients with primary-progressive MS), 10 people with RIS, and 47 control subjects (different diseases of the nervous system of a non-autoimmune or inflammatory nature). We showed a statistically significant increase in the content of various microbiota markers in the CSF of patients with MS compared with the control group. We found no change in the content of these markers in blood of patients with MS. This suggests a change of markers of microbial load at the level of the central nervous system, but not at the level of the whole organism. The greatest number of statistically significant differences with the control group was found in the content of markers in CSF of patients with MS in remission. In the acute stage, on the contrary, we found no statistically significant differences compared to the control group. In particular, in CSF of patients with MS in remission, a statistically significant increase in the content of bacterial plasmalogen (4.5 times), and increase in the level of microbial markers specific to Peptostreptococcus anaerobius, Pseudomonas aeruginosa, Eubacterium, Bifidobacterium, Butirivibrio, Moraxella, Acinetobacter, Propionibacterium acnes, as well as an increase of markers of the Epstein-Barr virus were found. In addition, there was an increase of campesterol, the likely source of which is campesterol-producing microfungi. In the CSF of subjects with RIS there were a statistically significant increase in the level of markers of the Epstein-Barr virus, Propionibacterium acnes, as well as Pseudomonas, Moraxella, and Acinetobacter. An association of MS with polymicrobial infection is possible. It is also likely that there is a certain pattern of increase of microbiota markers in the CSF of patients with MS, but not in blood.

4-Year Multiple Sclerosis Disease Remission After a Single Cycle of Alemtuzumab: a case report

Introduction. Alemtuzumab is a humanized monoclonal antibody against the CD52 antigen. It is a highly efficacious drug for the treatment of relapsing remitting multiple sclerosis (MS). The treatment protocol consists in 2 treatment cycles: 12mg/day i.v. infusions for 5 consecutive days and after one-year 12mg/day infusions for 3 days. To our knowledge, there is a single case series in the literature detailing MS remission after a single cycle of Alemtuzumab. Here, we report a similar experience. Case report. A 48-year-old woman with fluctuating right leg weakness for three years and burning pain in the right foot was diagnosed with MS. She was started on Betaferon©. Her condition progressed and in less than one year, she was switched to glatiramer and after new MS relapses to fingolimod. Disease progressed to Expanded Disability Status Scale (EDSS) of 7.0. She was started on Alemtuzumab (five-day course of 12mg i.v. on 6/2018). A second Alemtuzumab cycle was not given since she failed to follow monitoring guidelines. She had no signs of disease activity after four years. Conclusions. It is possible that MS remission may be achieved with a single cycle of Alemtuzumab.