Pathological Processes In Multiple Sclerosis Research Articles

Changes in <i>DNMT1</i> expression as a marker of epigenetic regulation disturbanses in multiple sclerosis patients

BACKGROUND: Multiple sclerosis is a chronic neurodegenerative autoimmune disease characterized by the presence of foci of inflammation and demyelination in the central nervous system. The initiation of pathological processes in multiple sclerosis is caused by a complex interaction of genetic factors, unfavorable environmental factors and epigenetic influences. Progressive neurological symptoms caused by axonal conduction disorders, axonal death and neurodestruction lead to a significant decreased patients’ quality of life and disability. The search for a new markers to improve diagnostic and therapeutic methods, including taking into account the genetic background and epigenetic interactions, is an urgent task.
 AIM: The work was aimed to study the changes in DNMT1 mRNA expression in multiple sclerosis patients with different disease duration, to analyze methylation of DNMT1 promoter, and compare the changes in the level of DNMT1 expression with the homocysteine content in the blood, and the presence of polymorphic variants in genes coding the key folate cycle enzymes.
 MATERIALS AND METHODS: The level of DNMT1 mRNA expression in peripheral mononuclear blood cells was assessed by reversed transcription followed by polymerase chain reaction. Fluorescent polymerase chain reaction followed by methyl-sensitive analysis of high-resolution melting curves was used to analyze methylation of the DNMT1 promoter. The content of homocysteine in the blood was determined by chemiluminescence immunoassay. The real-time polymerase chain reaction was used for genotyping by polymorphism of folate cycle genes; the fluorescent probes with the LNA modifications were used to discriminate alleles.
 RESULTS: It has been shown that in multiple sclerosis patients, including those at the onset of the disease, the level of DNMT1 mRNA expression is significantly lower than in the control group. No relationship was found between the decrease in DNMT1 expression and the level of promoter methylation. Strong positive relationship between the level of DNMT1 mRNA expression and homocysteine content in patients with multiple sclerosis and the combined effects of the genotypes of MTR A2756G and MTHFR C677T polymorphism on the expression of DNMT1 have been shown. These findings suggest that genetically determined features of folate metabolism may contribute to the disruption of epigenetic regulation in multiple sclerosis.
 CONCLUSIONS: The obtained results indicate the promise of research aimed to identifying the factors causing epigenetic changes in multiple sclerosis. Studying the mechanisms of the folate cycle genes polymorphic variants contribution to the pathogenesis of multiple sclerosis could be one of the possible ways to improve diagnostic and therapeutic approaches.

Effects of disease-modifying drugs in multiple sclerosis on the gut microbiome

Over the past decade, the scientific research has shown that the community of microorganisms that inhabit the gut, known as the gut microbiota, is closely linked to human health and disease, in part as a result of its influence on the systemic immune responses. The accumulated evidence suggests that these effects on the immune system are significant in neuroinflammatory diseases such as multiple sclerosis, and that modulation of the gut microbiota may be a potential therapeutic target in these conditions. In recent years, more and more studies have been appearing devoted to the role of microbiota in the development of various pathological processes in the central nervous system, including multiple sclerosis, through the brain-gut axis. In this regard, the question of finding ways to influence the microbiome is interesting. In addition to the existing attempts through the use of probiotics and fecal microbiota transplantation, of particular interest is the search for other ways of influencing the microbiome, including the effect of multiple sclerosis modifying therapies on the microbiota’s composition. The purpose of this review is to summarize the current evidence investigating the effects of disease-modifying treatment on the gut microbiome.

Background and roles: myosin in autoimmune diseases.

The myosin superfamily is a group of molecular motors. Autoimmune diseases are characterized by dysregulation or deficiency of the immune tolerance mechanism, resulting in an immune response to the human body itself. The link between myosin and autoimmune diseases is much more complex than scientists had hoped. Myosin itself immunization can induce experimental autoimmune diseases of animals, and myosins were abnormally expressed in a number of autoimmune diseases. Additionally, myosin takes part in the pathological process of multiple sclerosis, Alzheimer's disease, Parkinson's disease, autoimmune myocarditis, myositis, hemopathy, inclusion body diseases, etc. However, research on myosin and its involvement in the occurrence and development of diseases is still in its infancy, and the underlying pathological mechanisms are not well understood. We can reasonably predict that myosin might play a role in new treatments of autoimmune diseases.

Tyrosine kinases: a target of epigenetic influences and a new direction in the treatment of multiple sclerosis

Studies have shown that tyrosine kinases can be important in the pathogenesis of multiple sclerosis (MS) through the regulatory network of microRNAs. A microRNA network-based analysis revealed 17 receptor pathways activated by tyrosine kinase and regulated by microRNAs encoded at the DLK1-DIO3 locus on chromosome 14. Tyrosine kinases are actively involved in the epigenetic regulation of the pathological process in MS with the participation of a microRNA network and thus attract attention as a target for the development of new ways for the treatment of MS. The greatest attention was attracted by Bruton tyrosine kinase inhibitors (TKBi), which showed their ability to suppress the activity of autoimmune inflammatory lesions in the MS model - experimental autoimmune encephalomyelitis (EAE). This is due to the influence of TRB on the activity of B cells, which have a critical role in the development of the pathological process. The review discusses the types of TRBi, the features of their action in EAE and the results of the first studies in MS.

Lesion features on magnetic resonance imaging discriminate multiple sclerosis patients.

Magnetic resonance imaging (MRI) provides insight into various pathological processes in multiple sclerosis (MS) and may provide insight into patterns of damage among patients. We sought to determine if MRI features have clinical discriminative power among a cohort of MS patients. Ninety-six relapsing remitting and seven progressive MS patients underwent myelin water fraction (MWF) imaging and conventional MRI for cortical thickness and thalamic volume. Patients were clustered based on lesion level MRI features using an agglomerative hierarchical clustering algorithm based on principal component analysis (PCA). One hundred and three patients with 1689 MS lesions were analyzed. PCA on MRI features demonstrated that lesion MWF and volume distributions (characterized by 25th, 50th, and 75th percentiles) accounted for 87% of the total variability based on four principal components. The best hierarchical cluster confirmed two distinct patient clusters. The clustering features in order of importance were lesion median MWF, MWF 25th, MWF 75th, volume 75th percentiles, median individual lesion volume, total lesion volume, cortical thickness, and thalamic volume (all p values <0.01368). The clusters were associated with patient Expanded Disability Status Scale (EDSS) (n=103, p=0.0338) at baseline and at 5years (n=72, p=0.0337). These results demonstrate that individual MRI features can identify two patient clusters driven by lesion-based values, and our unique approach is an analysis blinded to clinical variables. The two distinct clusters exhibit MWF differences, most likely representing individual remyelination capabilities among different patient groups. These findings support the concept of patient-specific pathophysiological processes and may guide future therapeutic approaches.

Downregulation of miR-125a-5p and miR-218-5p in Peripheral Blood Mononuclear Cells of Patients with Relapsing-Remitting Multiple Sclerosis

ABSTRACT Multiple sclerosis (MS) is a chronic neuroinflammatory disease of the brain and spinal cord. Evidences have demonstrated that microRNAs (miRNAs) are involved in the pathological process of MS that may confer a valuable diagnostic biomarker for disease diagnosis, prognosis, and treatment. Hence, we assessed the expression pattern of miR-125a-5p and miR-218-5p in the peripheral blood mononuclear cells (PBMCs) of subjects with relapsing-remitting multiple sclerosis (RRMS). We recruited 50 RRMS patients and 50 age- and sex-matched healthy control subjects. PBMCs were isolated from the peripheral blood samples, RNA content was extracted, cDNA was synthesized, and finally expression level of miRNAs was determined using quantitative real-time PCR. Our data indicate significant downregulation of both miR-125a-5p and miR-218-5p in RRMS patients compared to healthy controls (P< .0001). The levels of both miRNAs were significantly downregulated in an age-dependent manner compared with consistent healthy control groups (30–40 years old P< .0001). Expression level of miR-218-5p was significantly changed in only female patients (Female group P< .0001; Male group P= .12). Receiver operating characteristic (ROC) curve data indicated that the expression levels of both miRNAs were able to discriminate RRMS patients from healthy subjects (P< .05). Moreover, bioinformatic enrichment analysis revealed that the target genes of these miRNAs had cardinal roles in the regulation of key biological pathways involved in the clinical course and pathogenesis of MS. Collectively, our results suggested that miR-125a-5p and miR-218-5p play a role in RRMS pathogenesis and have an age- and sex-dependent expression pattern in these patients.

Поражение подкорковых структур головного мозга при различных фенотипах рассеянного склероза и их прогностическая значимость

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease that affects the central nervous system, mainly in young people, and leads to inevitable disability. The purpose of the study is to determine the degree of involvement of subcortical structures in the pathological process in multiple sclerosis and to assess their prognostic significance. 80 patients with relapsing-remitting (n=48) and secondary progressive (n=32) phenotypes of MS were examined; the control group consisted of 20 healthy people of the corresponding age and gender. Clinical assessment was carried out on the following scales: EDSS, MSSS, MMSE, FAB, MoCA, SDMT, Beck’s test and HADS. All patients underwent MRI of the brain and MR-morphometry using the Freesurfer 6.0 software. In patients with multiple sclerosis, the neurodegenerative process is represented by a decrease in the volumes of the caudate nucleus and putamen, an increase in the volume of the third and lateral ventricles, an increase in CSF volume, and the presence of «black holes». The volume of the third and lateral ventricles, as well as the volume of CSF (general neurodegeneration), depends on the duration of the disease. The degree of disability (EDSS) is influenced by the volumes of the caudate nucleus, globus pallidus, nucleus accumbens, and brainstem. In turn, cognitive decline is influenced by the volume of the thalamus, basal nuclei, brainstem, the volume of the lateral ventricles, and a decrease in the volume of the white matter and cerebellar cortex. Thus, dynamic assessment and monitoring of the volume of subcortical structures of the brain using MR-morphometry can act as a prognostic factor in the transition of the relapsing-relapsing phenotype of multiple sclerosis to the secondary progressive phenotype. English version of this article on pp. 404-411 is available at URL: https://panor.ru/articles/subcortical-brain-lesions-in-different-phenotypes-of-multiple-sclerosis-and-their-prognostic-significance/63941.html

Evaluation of the expressed miR-129 and miR-549a in patients with multiple sclerosis.

Background:The expression of microRNAs (miRNAs) as circulating biomarkers has been underlined in multiple sclerosis (MS) in the last decade. Due to the presence of a possible relationship between expressed miRNAs and heterogeneous appearances of the pathological processes in MS, the present study attempts to evaluate the expression of miR-129 and miR-549a in patients with MS in comparison with healthy control (HC) group.Materials and Methods:Peripheral blood mononuclear cells were separated from fifty patients with MS (subtypes including relapsing–remitting MS and secondary progressive MS) in the Kashani Hospital, Isfahan, Iran, and fifty people as HC group. After RNA extraction and complementary DNA synthesis, the expression of miR-129 and miR-549a was evaluated in patients with MS in comparison with the HC group using a quantitative real-time polymerase chain reaction assay. The data were analyzed using the Kolmogorov–Smirnov and Mann–Whitney tests. Spearman's correlation coefficient was used to examine the relationship between miR-129 and miR-549a with age.Results:The results showed that the expression of miR-129 and miR-549a was not significant in patients with MS in comparison with the HC group. Furthermore, the relationship between such miRNAs and age and gender was not significant.Conclusion:We suggest the expression of miR-129 and miR-549a as circulating miRNAs in peripheral blood mononuclear cells could not be considered a biomarker for diagnosis and Para clinical.

Bu-Shen-Yi-Sui Capsule, an Herbal Medicine Formula, Promotes Remyelination by Modulating the Molecular Signals via Exosomes in Mice with Experimental Autoimmune Encephalomyelitis.

Multiple sclerosis (MS) is a common inflammatory demyelinating disorder of the central nervous system. Bu-shen-yi-sui capsule (BSYSC) could significantly reduce the relapse rate, prevent the progression of MS, and enhance remyelination following neurological injury in experimental autoimmune encephalomyelitis (EAE), an established model of MS; however, the mechanism underlying the effect of BSYSC on remyelination has not been well elucidated. This study showed that exosomes carrying biological information are involved in the pathological process of MS and that modified exosomes can promote remyelination by modulating related proteins and microRNAs (miRs). Here, the mechanism by which BSYSC promoted remyelination via exosome-mediated molecular signals was investigated in EAE mice and oligodendrocyte progenitor cells (OPCs) in vitro. The results showed that BSYSC treatment significantly improved the body weight and clinical scores of EAE mice, alleviated inflammatory infiltration and nerve fiber injury, protected the ultrastructural integrity of the myelin sheath, and significantly increased the expression of myelin basic protein (MBP) in EAE mice. In an in vitro OPC study, BSYSC-containing serum, especially 20% BSYSC, promoted the proliferation and migration of OPCs and induced OPCs to differentiate into mature oligodendrocytes that expressed MBP. Furthermore, BSYSC treatment regulated the expression of neuropilin- (NRP-) 1 and GTX, downregulated the expression of miR-16, let-7, miR-15, miR-98, miR-486, and miR-182, and upregulated the level of miR-146 in serum exosomes of EAE mice. In conclusion, these results suggested that BSYSC has a neuroprotective effect and facilitates remyelination and that the mechanism underlying the effect of BSYSC on remyelination probably involves regulation of the NRP-1 and GTX proteins and miRs in serum exosomes, which drive promyelination.

Myelin water imaging from multi-echo [formula omitted] MR relaxometry data using a joint sparsity constraint

Demyelination is the key pathological process in multiple sclerosis (MS). The extent of demyelination can be quantified with magnetic resonance imaging by assessing the myelin water fraction (MWF). However, long computation times and high noise sensitivity hinder the translation of MWF imaging to clinical practice. In this work, we introduce a more efficient and noise robust method to determine the MWF using a joint sparsity constraint and a pre-computed B1+-T2 dictionary.A single component analysis with this dictionary is used in an initial step to obtain a B1+ map. The T2 distribution is then determined from a reduced dictionary corresponding to the estimated B1+ map using a combination of a non-negativity and a joint sparsity constraint.The non-negativity constraint ensures that a feasible solution with non-negative contribution of each T2 component is obtained. The joint sparsity constraint restricts the T2 distribution to a small set of T2 relaxation times shared between all voxels and reduces the noise sensitivity.The applied Sparsity Promoting Iterative Joint NNLS (SPIJN) algorithm can be implemented efficiently, reducing the computation time by a factor of 50 compared to the commonly used regularized non-negative least squares algorithm. The proposed method was validated in simulations and in 8 healthy subjects with a 3D multi-echo gradient- and spin echo scan at 3 ​T. In simulations, the absolute error in the MWF decreased from 0.031 to 0.013 compared to the regularized NNLS algorithm for SNR ​= ​250. The in vivo results were consistent with values reported in literature and improved MWF-quantification was obtained especially in the frontal white matter. The maximum standard deviation in mean MWF in different regions of interest between subjects was smaller for the proposed method (0.0193) compared to the regularized NNLS algorithm (0.0266). In conclusion, the proposed method for MWF estimation is less computationally expensive and less susceptible to noise compared to state of the art methods. These improvements might be an important step towards clinical translation of MWF measurements.

Effects of guluronic acid (G2013) on gene expression of TLR2, TLR4, MyD88, TNF-α and CD52 in multiple sclerosis under in vitro conditions

Context: Multiple sclerosis (MS) is an autoimmune and chronic inflammatory disease of CNS. The α-L-guluronic acid (G2013) as novel NSAID with immunomodulatory effects has shown its positive effects in various investigations.Objective: Present research aimed to study the potency of G2013 on gene expression of TLR2, TLR4, MyD88, TNF-α and CD52 in PBMCs of MS patients under in vitro conditions. Materials and methods: 24 blood samples from MS patients and healthy controls were considered for RT-PCR and flow cytometry techniques under two different doses of G2013.Results: Our research indicated that this drug could significantly decrease the gene expression of TLR2, TLR4 and TNF-α compared to untreated group. Conclusion: Data demonstrated that the guluronic acid is able to modify the expression levels of TLR2, TLR4 and TNF-α genes to less than the pathogenic boarder line level, which it might be recommended for reducing the pathological process in multiple sclerosis.

Potential of Adult Endogenous Neural Stem/Progenitor Cells in the Spinal Cord to Contribute to Remyelination in Experimental Autoimmune Encephalomyelitis.

Demyelination and remyelination play pivotal roles in the pathological process of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), a well-established animal model of MS. Although increasing evidence shows that various stimuli can promote the activation/induction of endogenous neural stem/progenitor cells (NSPCs) in the central nervous system, the potential contributions of these cells to remyelination following inflammatory injury remain to be fully investigated. In the present study, using an adult mouse model of EAE induced by myelin oligodendrocyte glycoprotein (MOG) peptide, we investigated whether adult NSPCs in the spinal cord can lead to remyelination under inflammatory conditions. Immunohistochemistry showed that cells expressing the NSPC marker Nestin appeared after MOG peptide administration, predominantly at the sites of demyelination where abundant inflammatory cells had accumulated, whereas Nestin+ cells were rarely present in the spinal cord of PBS-treated control mice. In vitro, Nestin+ NSPCs obtained from EAE mice spinal cords could differentiate into multiple neural lineages, including neurons, astrocytes, and myelin-producing oligodendrocytes. Using the Cre–LoxP system, we established a mouse strain expressing yellow fluorescent protein (YFP) under the control of the Nestin promoter and investigated the expression patterns of YFP-expressing cells in the spinal cord after EAE induction. At the chronic phase of the disease, immunohistochemistry showed that YFP+ cells in the injured regions expressed markers for various neural lineages, including myelin-forming oligodendrocytes. These results show that adult endogenous NSPCs in the spinal cord can be subject to remyelination under inflammatory conditions, such as after EAE, suggesting that endogenous NSPCs represent a therapeutic target for MS treatment.

Associations of Optical Coherence Tomography Parameters and Pathological Process in Multiple Sclerosis: Evidence from a Systematic Review and Meta-Analysis

Associations of Optical Coherence Tomography Parameters and Pathological Process in Multiple Sclerosis: Evidence from a Systematic Review and Meta-Analysis

Pro-Thrombotic Activity of Blood Platelets in Multiple Sclerosis.

The available data, including experimental studies, clearly indicate an excessive intravascular activation of circulating platelets in multiple sclerosis (MS) and their hyper-responsiveness to a variety of physiological activators. Platelet activation is manifested as an increased adhesion and aggregation and is accompanied by the formation of pro-thrombotic microparticles. Activated blood platelets also show an expression of specific membrane receptors, synthesis many of biomediators, and generation of reactive oxygen species. Epidemiological studies confirm the high risk of stroke or myocardial infarction in MS that are ischemic incidents, strictly associated with incorrect platelet functions and their over pro-thrombotic activity. Chronic inflammation and high activity of pro-oxidative processes in the course of MS are the main factors identified as the cause of excessive platelet activation. The primary biological function of platelets is to support vascular integrity, but the importance of platelets in inflammatory diseases is also well documented. The pro-thrombotic activity of platelets and their inflammatory properties play a part in the pathophysiology of MS. The analysis of platelet function capability in MS could provide useful information for studying the pathogenesis of this disease. Due to the complexity of pathological processes in MS, medication must be multifaceted and blood platelets can probably be identified as new targets for therapy in the future.

Systemic Acrolein Elevations in Mice With Experimental Autoimmune Encephalomyelitis and Patients With Multiple Sclerosis.

Demyelination and axonal injury are the key pathological processes in multiple sclerosis (MS), driven by inflammation and oxidative stress. Acrolein, a byproduct and instigator of oxidative stress, has been demonstrated as a neurotoxin in experimental autoimmune encephalomyelitis (EAE), an animal model of MS. However, due to the invasive nature of acrolein detection using immunoblotting techniques, the investigation of acrolein in MS has been limited to animal models. Recently, detection of a specific acrolein-glutathione metabolite, 3-HPMA, has been demonstrated in urine, enabling the noninvasive quantification of acrolein for the first time in humans with neurological disorders. In this study, we have demonstrated similar elevated levels of acrolein in both urine (3-HPMA) and in spinal cord tissue (acrolein-lysine adduct) in mice with EAE, which can be reduced through systemic application of acrolein scavenger hydralazine. Furthermore, using this approach we have demonstrated an increase of 3-HPMA in both the urine and serum of MS patients relative to controls. It is expected that this noninvasive acrolein detection could facilitate the investigation of the role of acrolein in the pathology of MS in human. It may also be used to monitor putative therapies aimed at suppressing acrolein levels, reducing severity of symptoms, and slowing progression as previously demonstrated in animal studies.

Features of the Course of the Pathological Process in Multiple Sclerosis as Confirmed by Changes in Biochemical Markers for Oxidative Stress

Objective. To analyze the features of the course of the pathological process in multiple sclerosis (MS) in terms of measures of oxidative stress and antioxidant defense (AOD). Materials and methods. A total of 220 patients with different clinical forms and stages of MS were studied; patients were divided into four clinical groups. Markers of the intensity of free radical oxidation of lipids and proteins were evaluated in terms of the levels of ketodienes, diene conjugates, malondialdehyde, and Schiff bases. The functioning of the endogenous AOD was assessed in terms of measures of the activity of the following markers: vitamin E and SH groups – total and nonprotein thiols – to assess the nonenzymatic compartment of the AOD. Markers of the enzymatic compartment were: superoxide dismutase, glutathione peroxidase, and glutathione reductase. Results and conclusions. Differences were found in measures of the intensity of oxidative stress in groups of patients with different clinical forms and stages of MS, not only verifying oxidative stress in MS from the point of view of evidence-based medicine, but also supporting the continuity of the pathological process in this disease.

Longitudinal MR spectroscopy of neurodegeneration in multiple sclerosis with diffusion of the intra-axonal constituent N-acetylaspartate

Multiple sclerosis (MS) is a pathologically complex CNS disease: inflammation, demyelination, and neuroaxonal degeneration occur concurrently and may depend on one another. Current therapies are aimed at the immune-mediated, inflammatory destruction of myelin, whereas axonal degeneration is ongoing and not specifically targeted. Diffusion-weighted magnetic resonance spectroscopy can measure the diffusivity of metabolites in vivo, such as the axonal/neuronal constituent N-acetylaspartate, allowing compartment-specific assessment of disease-related changes. Previously, we found significantly lower N-acetylaspartate diffusivity in people with MS compared to healthy controls (Wood et al., 2012) suggesting that this technique can measure axonal degeneration and could be useful in developing neuroprotective agents. In this longitudinal study, we found that N-acetylaspartate diffusivity decreased by 8.3% (p < 0.05) over 6 months in participants who were experiencing clinical or MRI evidence of inflammatory activity (n = 13), whereas there was no significant change in N-acetylaspartate diffusivity in the context of clinical and radiological stability (n = 6). As N-acetylaspartate diffusivity measurements are thought to more specifically reflect the intra-axonal space, these data suggest that N-acetylaspartate diffusivity can report on axonal health on the background of multiple pathological processes in MS, both cross-sectionally and longitudinally.

Heterogeneity of acute multiple sclerosis lesions on sodium (23Na) MRI

Background: Advanced magnetic resonance imaging (MRI) techniques provide a window into pathological processes in multiple sclerosis (MS). Nevertheless, to date only few studies have performed sodium MRI in MS. Objectives: We analysed total sodium concentration (TSC) in hyperacute, acute and chronic lesions in MS with 23Na MRI. Methods: 23Na MRI and 1H MRI were performed in 65 MS patients and 10 healthy controls (HC). Mean TSC was quantified in all MS lesions with a diameter of >5 mm and in the normal appearing white and grey matter (NAWM, NAGM). Results: TSC in the NAWM and the NAGM of MS patients was significantly higher compared to HC (WM: 37.51 ± 2.65 mM versus 35.17 ± 3.40 mM; GM: 43.64 ± 2.75 mM versus 40.09 ± 4.64 mM). Acute and chronic MS lesions showed elevated TSC levels of different extent (contrast-enhancing lesions (49.07 ± 6.99 mM), T1 hypointense lesions (45.06 ± 6.26 mM) and remaining T1 isointense lesions (39.88 ± 5.54 mM)). However, non-enhancing hyperacute lesions with a reduced apparent diffusion coefficient showed a TSC comparable to the NAWM (37.22 ± 4.62 mM). Conclusions: TSC is not only a sensitive marker of the severity of chronic tissue abnormalities in MS but is also highly sensitive to opening of the blood–brain barrier and vasogenic tissue oedema in contrast-enhancing lesions.

Multiparameter Flow Cytometric Assays to Quantify Effector and Regulatory T-Cell Function in Multiple Sclerosis.

The immune system plays a major pathological and regulatory role in multiple sclerosis (MS) and, therefore, is a focus of extensive research. Animal models of MS have been crucial in understanding the pathological processes in MS and developing certain treatments, however, all crucial aspects of the human disease may not be appropriately modeled. With the exception of detecting oligoclonal bands and IgG synthesis in cerebrospinal fluids of MS patients, there has not been major progress in the development of immunologic tests that can be used for diagnosis of MS. Further, due to the lack of validated immune assays, routine monitoring of the immune system following therapy initiation is not a part of standard patient care in MS. This is critical since immunomodulatory therapies used for MS treatment are not benign and, more importantly, there is a considerable variation in clinical responses in MS patients initiating such therapies. Flow cytometry is a powerful tool that can be used for studying both the phenotype and function of immune cells. The studies described here will demonstrate how flow cytometry can be used to apply current knowledge about the MS immune system to develop a diagnostic laboratory test for the immunologic monitoring of this disease. Importantly, we will also show that the multiparameter flow cytometry based assay developed by us can also be implemented for the immunologic evaluation of therapeutic success in MS patients.

Oxysterols and cholesterol precursors correlate to magnetic resonance imaging measures of neurodegeneration in multiple sclerosis

Background: Cholesterol homeostasis is important for formation and maintenance of myelin and axonal membranes in the central nervous system (CNS). The concentrations of the brain specific cholesterol metabolite 24S-hydroxycholesterol (24OHC) and cholesterol precursors have been shown to be altered in multiple sclerosis (MS). However, how changes in sterol levels relate to the pathological processes in MS is not clear. Methods: In this study, we compared serum and cerebrospinal fluid (CSF) sterol levels between 105 MS (51 relapsing–remitting (RR); 39 secondary progressive (SP) and 15 primary progressive (PP)) and 49 control patients. Sterol levels were correlated to magnetic resonance imaging (MRI) markers of disease activity. Results: We found decreased serum 24OHC and 27-hydroxycholesterol (27OHC) and increased CSF lathosterol in MS patients compared to control patients (p=0.018, p=0.002 and p=0.002, respectively). Subgroup analysis showed that serum 24OHC levels were negatively correlated to normalized brain volume measurements in relapse-onset MS patients (r= −0.326, p=0.004). Conclusions: These results confirm that cholesterol homeostasis is disturbed in MS and suggest that changes in cholesterol synthesis are related to neurodegenerative pathological processes as seen on the MRI. The data seem to be in line with the recently reported observation that high dose statins may have a positive effect on clinical disability in secondary progressive MS.