Other Neurological Diseases Research Articles

Urinary Extracellular Domain of Neurotrophin Receptor p75 as a Biomarker for Amyotrophic Lateral Sclerosis in a Chinese cohort

To comprehensively assess whether p75ECD in urine could be a candidate biomarker for ALS evaluation. Urine samples were collected from 101 ALS patients, 108 patients with other neurological disease (OND) and 97 healthy controls. 61 ALS patients were followed up with clinical data including ALSFRS-r every 6 to 12 months, 23 ALS patients died and 17 ALS patients lost touch during follow up period. Enzyme-linked immunoassay was employed to determine urine p75ECD concentration. The ALSFRS-r was employed to assess the severity of ALS. The concentration of p75ECD in ALS was significantly higher than that of OND and CTRL (p < 0.001). Additionally, urine p75ECD concentrations in ALS-definite grade patients were significantly higher than that in ALS-probable grade and ALS-possible grade patients (p < 0.001). Higher urine p75ECD concentrations were correlated with increased clinical stage (p = 0.0309); urine p75ECD concentrations and ALSFRS-r were negatively correlated (p = 0.022); and urine p75ECD concentration in the fast-progressing ALS group was significantly higher than that in slow-progression (p = 0.0026). Our finding indicates that urine p75ECD concentration provides additional evidence for patients with clinically suspected ALS, and can be employed to evaluate ALS-severity.

Multimarker risk stratification approach at multiple sclerosis onset

Delay in the diagnosis of multiple sclerosis (MS) stems from the lack of specific clinical and analytical markers to assist in the early diagnosis and prediction of progressive course. We propose a decision-tree model that better defines early at onset MS patients and those with the progressive form by analysing a 12-biomarkers panel in serum and CSF samples of patients with MS, other neurological diseases (OND) and healthy contols. Thus, patients at onset of neurological disease were first classified by serum IL-7 levels <141pg/ml (OR=6.51, p<0.001). Combination of IL-7 and CXCL10 indicated risk for a specific MS clinical form, where IL-7<141 and CXCL10<570pg/ml were associated with the highest risk for PP-MS (OR=22, p=0.01). Unexpectedly, both PP-MS and RR-MS patients shared significantly decreased prototypical biomarkers of inflammation and tissue regeneration in CSF than OND suggesting a defective intrinsic immune response playing a role at the beginning of the disease.

Detection and clinical significance of serum autoantibodies in patients with myasthenia gravis

Objective To investigate the expressions and clinical significance of serum anti - acetylcholine receptor antibodies (AChR-Ab), anti-Titin antibodies (Titin-Ab), and anti-Ryanodine receptor antibodies (RyR-Ab) in patients with myasthenia gravis (MG). Methods Serum AChR-Ab, Titin-Ab and RyR-Ab were detected with enzyme-linked immunosorbent assay (ELISA) in 182 MG patients, 105 patients of other neurological diseases (OND) and 62 normal controls. Results Serum AChR-Ab, Titin-Ab and RyR-Ab were detected positive respectively in 68.13% (124/182), 64.29% (117/182) and 67.03% (122/182) of patients in MG group. The positive rates of those antibodies in MG group were significantly higher than those in OND group ( P = 0.000, for all) and control group ( P = 0.000, for all). When 3 antibodies coexisted, the sensitivity in the diagnosis of MG was 41.21%, with 99.40% of specificity. There was no significant difference in the positive rate of serum AChR-Ab, Titin-Ab and RyR-Ab between early-onset MG subgroup and late-onset MG subgroup ( P > 0.05, for all). The positive rates of AChR-Ab were significantly higher in MG patients with thymoma than in those without thymoma ( P = 0.004). There was no significant difference in the positive rate of Titin-Ab and RyR-Ab between MG with thymoma subgroup and MG without thymoma subgroup ( P > 0.05, for all). The prevalence of AChR-Ab in generalized MG (GMG) atients (Ⅱa and Ⅱb) was higher than those in ocular MG (OMG) patients (typeⅠ; P = 0.005, 0.012). There was no significant difference in the positive rate of Titin-Ab and RyR-Ab between GMG subgroup and OMG subgroup ( P > 0.05, for all). Conclusions Serum AChR-Ab, Titin-Ab and RyR-Ab can be used as ndicators of the diagnosis of MG. Patients with 3 coexisted positive antibodies are highly suspected as MG. Higher AChR-Ab level in serum of OMG patients indicates the possibility of progressing to GMG. DOI: 10.3969/j.issn.1672-6731.2016.10.007

No Definite Evidence for Human Endogenous Retroviral HERV-W and HERV-H RNAS in Plasma of Latvian Patients Suffering from Multiple Sclerosis and Other Neurological Diseases / Nav Apstiprinošu Pierādījumu Par Endogēno Retrovīrusu HERV-W Un HERV-H Rns Sastopamību Latvijas Pacientu Ar Multiplo Sklerozi Un Citām NeiroloĢiskām Slimībām Asins Plazmā

Abstract Multiple sclerosis (MS) is a neurological disease of unknown aetiology. Several research groups reported an increased level of human endogenous retroviruses HERV-W and HERV-H RNAs in cerebrospinal fluid, plasma and supernatants of cell cultures from MS individuals. To quantify the abundance of extracellular virion-associated HERV, RNAs in blood, plasma samples from Latvian MS patients, patients with other neurological diseases (OND), as well as blood donors (BD), were retrospectively studied by using both our previously published and newly developed quantitative Real-time reverse transcription PCR assays (QPCRs) targeting different polymerase (pol) gene regions of HERV-W and HERV-H. Unspecific signals due to incomplete removal of DNA were monitored by running the assays with and without reverse transcription (RT±) in parallel. According to our score, a few MS, OND and healthy controls gave borderline signals simultaneously with both newly developed HERV-H and HERV-W QPCRs, but the rest were negative. All borderline positive samples also had small amounts of non-retroviral cellular mRNA with possible origin from cell-free circulating RNA fragments, apoptotic bodies or exosomes, which can mimic the previously described virus-like particles. The results do not confirm the previous reports on prevalence of HERV-H or -W virion-associated RNA in plasma of MS patients.

Serum BAFF levels, Methypredsinolone therapy, Epstein-Barr Virus and Mycobacterium avium subsp. paratuberculosis infection in Multiple Sclerosis patients.

Elevated B lymphocyte activating factor BAFF levels have been reported in multiple sclerosis (MS) patients; moreover, disease-modifying treatments (DMT) have shown to influence blood BAFF levels in MS patients, although the significance of these changes is still controversial. In addition, BAFF levels were reported increased during infectious diseases. In our study, we wanted to investigate on the serum BAFF concentrations correlated to the antibody response against Mycobacterium avium subspecies paratuberculosis (MAP), Epstein-Barr virus (EBV) and their human homologous epitopes in MS and in patients affected with other neurological diseases (OND), divided in Inflammatory Neurological Diseases (IND), Non Inflammatory Neurological Diseases (NIND) and Undetermined Neurological Diseases (UND), in comparison to healthy controls (HCs). Our results confirmed a statistically significant high BAFF levels in MS and IND patients in comparison to HCs but not NIND and UND patients. Interestingly, BAFF levels were inversely proportional to antibodies level against EBV and MAP peptides and the BAFF levels significantly decreased in MS patients after methylprednisolone therapy. These results implicate that lower circulating BAFF concentrations were present in MS patients with humoral response against MAP and EBV. In conclusion MS patients with no IgGs against EBV and MAP may support the hypothesis that elevated blood BAFF levels could be associated with a more stable disease.

Ganglionic acetylcholine receptor autoantibodies in patients with Guillain-Barré syndrome

ObjectivesAlthough standardized autonomic tests are useful for diagnosing autonomic failure in patients with Guillain-Barré syndrome (GBS), they cannot be used as predictive markers. Thus, serological markers may correctly identify patients with GBS who are at risk for autonomic dysfunction. MethodsWe validated a luciferase immunoprecipitation system that detects IgG antibodies in patient serum that specifically bind to the α3 or β4 subunits of ganglionic neuronal nicotinic acetylcholine receptors (gAChR). We then used luciferase-conjugated ligands specific to antibodies against two gAChR subunits to test 79 sera samples from patients with GBS, 34 from subjects with other neurological diseases (OND), and 73 from healthy controls (HC). 1) In the first analysis, patients were classified into two groups according to the presence or absence of autonomic symptoms (AS). We compared the frequency of the anti-gAChR antibodies between these two groups (AS+ and AS−). 2) In the second analysis, furthermore, patients were classified depending on the presence or absence of anti-glycolipid antibodies (AGA). We compared the frequency of the anti-gAChR antibodies between the four categories of GBS (AS+/AGA+, AS+/AGA−, AS−/AGA+, and AS−/AGA−), OND, and HC. ResultsEight subjects with GBS were positive for α3 subunits, while one was positive for β4 subunits. Anti-α3 and -β4 gAChR antibodies were also detected in 13.6% of AS+ GBS group in the first analysis. Two of 35 patients in AS−GBS group were seropositive for the anti-gAChR antibodies and AGA in the second analysis. Patients with GBS that were positive for serum antibodies to the α3 and/or β4 subunits of gAChRs showed a range of clinical features including AS and AGA. ConclusionsPatients with GBS may have circulating antibodies against gAChR, which may contribute to the autonomic dysfunction associated with this disease.

Proteomic analysis of cerebrospinal fluid in amyotrophic lateral sclerosis.

The present study used comparative proteomic analysis of cerebrospinal fluid (CSF) in amyotrophic lateral sclerosis (ALS) patients in order to identify proteins that may act as diagnostic biomarkers and indicators of the pathogenesis of ALS. This analysis was performed using isobaric tags for relative and absolute quantitation (iTRAQ) technology, coupled with 2-dimensional liquid chromatography/mass spectrometry. Database for Annotation, Visualization and Integrated Discovery software was utilized for bioinformatic analysis of the data. Following this, western blotting was performed in order to examine the expression of 3 candidate proteins in ALS patients compared with healthy individuals [as a normal control (NC) group] or patients with other neurological disease (OND); these proteins were insulin-like growth factor II (IGF-2), glutamate receptor 4 (GRIA4) and leucine-rich α-2-glycoprotein 1 (LRG1). Clinical data, including gender, age, disease duration and ALS functional rating scale (ALSFRS-R) score, were also collected in the ALS patients. Multiple linear regression analysis was performed between the clinical data and the results of western blot analysis. A total of 248 distinct proteins were identified in the ALS and NC groups, amongst which a significant difference could be identified in 35 proteins; of these, 21 proteins were downregulated and 14 were upregulated. These differentially-expressed proteins were thus revealed to be associated with ALS. The western blot analysis confirmed a proportion of the data attained in the iTRAQ analysis, revealing the differential protein expression of IGF-2 and GRIA4 between the ALS and NC groups. IGF-2 was significantly downregulated in ALS patients (P=0.017) and GRIA4 was significantly upregulated (P=0.016). These results were subsequently validated in the 35-patient ALS and OND groups (P=0.002), but no significant difference was identified in LRG1 expression between these groups. GRIA4 protein expression was higher in male than female patients and was positively correlated with the ALSFRS-R score, meaning that GRIA4 expression was negatively correlated with the severity of ALS, while IGF-2 and LRG1 expression did not correlate with any clinical data. The present study thus demonstrated that GRIA4 expression levels, as a marker of severity, may be used as a reference for the timing of treatment, and that IGF-2 may serve as an effective biomarker of ALS progression.

Epstein Barr Virus and Mycobacterium avium subsp. paratuberculosis peptides are recognized in sera and cerebrospinal fluid of MS patients.

Mycobacterium avium subsp. paratuberculosis (MAP) and Epstein-Barr virus (EBV) epitopes elicit a consistent humoral response in serum of multiple sclerosis patients, but the cross reactivity against the homologous myelin basic protein (MBP) and human interferon regulatory factor 5 (IRF5) has not been searched within the Cerebral Spinal Fluid (CSF). We evaluated in sera and CSF of patients with MS and with other neurological diseases (OND) the humoral response against EBV/MAP peptides and the IRF5/MBP. Our data showed that EBV and MAP peptides are able to induce a specific humoral immune response in MS patients compared to OND controls both in serum and in CSF. An intrathecal specific synthesis of IgG against MBP and their EBV and MAP homologous as indicated by the antibody index was observed in MS patients. The humoral response against EBV, MAP, MBP and IRF5 was significantly higher in MS patients compared to OND both in serum and in CSF. The higher presence of antibodies against MBP and their MAP and EBV homologous in CSF during relapses suggests a possible role of the pathogens in enhancing inflammation.

Humoral response to EBV is associated with cortical atrophy and lesion burden in patients with MS.

Objective:Because dysregulated Epstein-Barr virus (EBV)-infected B cells may induce meningeal inflammation, which contributes to cortical pathology in multiple sclerosis (MS), we investigated associations between antibody responses to EBV and development of cortical pathology in MS.Methods:We included 539 patients with MS (369 with relapsing-remitting MS, 135 with secondary progressive MS, and 35 with primary progressive MS), 66 patients with clinically isolated syndrome (CIS), 63 patients with other neurologic diseases (OND), and 178 age- and sex-matched healthy controls (HC). All participants were scanned on 3T MRI. Serum samples were analyzed for IgG antibodies against EBV viral capsid antigen (VCA) and EBV nuclear antigen-1 (EBNA-1), and their quartiles were determined on the whole study sample. Differences between the study groups were assessed using analysis of covariance adjusted for multiple comparisons.Results:More than 30% of patients with MS and CIS presented with the highest quartile of anti-EBV-VCA and -EBNA-1 status compared to ≤10% of HC (p < 0.001). The figures were 9 (14.3%) and 7 (12.3%) for patients with OND. Patients with MS with the highest quartile of anti-EBV-VCA showed significantly increased T2 lesion volume (p = 0.001), T1 lesion number (p = 0.002), and T1 lesion volume (p = 0.04) and decreased gray matter (p = 0.041) and cortical (p = 0.043) volumes compared to patients with MS with lower quartiles. No significant differences of MRI outcomes in patients with CIS, patients with OND, and HC with lower or highest quartiles of anti-EBV-VCA and -EBNA-1 were detected.Conclusions:Humoral response to anti-EBV-VCA and -EBNA-1 is associated with more advanced cortical atrophy, accumulation of chronic T1 black holes, and focal white matter lesions in patients with MS.

Quantified CSF antibody reactivity against myelin in multiple sclerosis.

Synthesis of clonal IgG is a consistent feature of patients with multiple sclerosis (MS). Whether oligoclonal bands (OCBs) represent unspecific disease bystanders or active components in MS pathology is an open question. The aim of this study was to develop a method to quantify and compare the reactivity of cerebrospinal fluid (CSF) antibodies from patients with and without MS. We collected CSF from 262 patients from two different cohorts, which included 148 patients with MS and 114 with other neurological diseases (OND). We established a highly sensitive electrochemiluminescence (ECL)-based assay to measure CSF antibody reactivity against purified myelin particles and biotin anchored liposomes. The diagnostic value of the ECL score against myelin particles was assessed with receiver operating characteristic curves. CSF from patients with MS have higher reactivity toward purified myelin particles as compared to those with OND with OCBs. Using liposomes with defined lipid compositions and myelin particles from ceramide synthase 2 (CerS2) knockout mice, we find that some of the CSF antibody reactivity is directed against cerebrosides. The ECL-based assay system expands the currently available toolbox for the detection of autoantibodies in MS and related diseases.

Label-Free LC-MS/MS Proteomic Analysis of Cerebrospinal Fluid Identifies Protein/Pathway Alterations and Candidate Biomarkers for Amyotrophic Lateral Sclerosis.

Analysis of the cerebrospinal fluid (CSF) proteome has proven valuable to the study of neurodegenerative disorders. To identify new protein/pathway alterations and candidate biomarkers for amyotrophic lateral sclerosis (ALS), we performed comparative proteomic profiling of CSF from sporadic ALS (sALS), healthy control (HC), and other neurological disease (OND) subjects using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 1712 CSF proteins were detected and relatively quantified by spectral counting. Levels of several proteins with diverse biological functions were significantly altered in sALS samples. Enrichment analysis was used to link these alterations to biological pathways, which were predominantly related to inflammation, neuronal activity, and extracellular matrix regulation. We then used our CSF proteomic profiles to create a support vector machines classifier capable of discriminating training set ALS from non-ALS (HC and OND) samples. Four classifier proteins, WD repeat-containing protein 63, amyloid-like protein 1, SPARC-like protein 1, and cell adhesion molecule 3, were identified by feature selection and externally validated. The resultant classifier distinguished ALS from non-ALS samples with 83% sensitivity and 100% specificity in an independent test set. Collectively, our results illustrate the utility of CSF proteomic profiling for identifying ALS protein/pathway alterations and candidate disease biomarkers.

Clinical practice and decision making in as

Background: Seric and urinary markers are increased in Acute Strokes (AS) and Chronic Cerebrovascular Diseases (CCVD) compared to Other Neurological Diseases (OND) (intergroup variability). No significant fluctuations were found within each group of patients at repeated measurements (intragroup unvariability). Objective: The aim of our current study is to evaluate the predictive values and the correlations with clinical, echographic and radiological findings. Patients and Methods: So far, we recruited 129 OND, 398 CCVD, 483 AS. We classified them in subgroups according to the severity of neurological and heart dysfunctions. Results: The most significant alterations of cardiac and urinary markers were detected in AS in class III/C, IV/C and IV/D of New York Heart Association / American Cardiology Association scales, especially in cases with concomitant CCVD, and in unstable CCVD. No significant intragroup differences were found at repeated measurements. CRP, Tro ths, NT-pro-BNP, proteinuria had predictive values. Correlations were found with CHAD2DS2VAsc, HAS BLED, Hachinski, Apache, Glasgow Coma Scale, Glasgow Outcome Scale, Modified Rankin Scale, echocardiographic parameters, Simplified Pulmonary Embolism Severity and Pulmonary Embolism Severity Indices. Conclusions: Our data highlight important features within the same category of AS accounting for worst outcomes, restricting therapeutical effectiveness, prolonging hospitalization and predicting bounce backs, above all in patients affected with CCVD and severe cardiac dysfunctions. While more sophisticated radiological techniques may show a continuum from physiological to subtle pathological conditions, the other parameters allow early identifying and treating emergencies. Computerized modeling would monitor decision making and better define the burden, reversibility and lesional load of cerebrovascular disease.

High frequency of Human Polyomavirus 6 DNA in the cerebrospinal fluid of patients with neurological diseases

No: 1743 Presentation at ESCV 2015: Poster 1 High frequency of Human Polyomavirus 6 DNA in the cerebrospinal fluid of patients with neurological diseases L. Signorini1,∗, F. Elia1, S. Villani1, R. Ticozzi1, M. Dolci1, D. Fraciotta2, E. Marchioni2, P. Ferrante1, S. Delbue1 1 University of Milano, Italy 2 Istituto Neurologico Mondino, Pavia, Italy Background: The family Polyomaviridae comprises thirteen human viruses that have been detected in various specimen types, mainly urine, skin and blood. Among all of them, only JC Polyomavirus (JCPyV), and sporadically BK polyomavirus (BKPyV) genomes have been detected in the Cerebrospinal Fluid (CSF). More recently, the unexpected finding of Human Polyomavirus 6 (HPyV6) DNA in the CSF from anHIV positive patientwith leukoencephalopathy was reported. JCPyV being a recognized cause of Progressive Multifocal Leukoencephalopathy (PML) in immunodeficient subjects provides a precedent for polyomavirus causing neurological symptoms, but alternative explanations should also be considered. The objectives of the study were to explore the prevalence of the newly discovered HPyV6, Human Polyomavirus 7 (HPyV7) andHuman Polyomavirus 9 (HPyV9) in neurological diseases, in comparison with those of JCV. Methods:CSFhavebeen collected from51HIV-positivepatients affected with HIV-related leukoencephalopathies and 191 HIVnegative patients affected with other neurological diseases (OND), such as meningitis, encephalitis, and encephalomyelitis. DNA was isolated and real-timePCRassays for JCV,HPyV6,HPyV7andHPyV9 were conducted. Results: JCV genome was detected in the CSF of 16/51 (31.4%) HIV-related leukoencephalopathies patients, HPyV6 genome in the CSF of 2/51 (3.9%) HIV-related leukoencephalopathies patients and in 37/191 (19.4%) HIV-negative patients with OND. HPyV7 and HPyV9 genomes were not amplified in any clinical specimens. Conclusions: HPyV6 genome was found in a high percentage of the CSF of the OND patients, whereas HPyV7 and HPyV9 genomeswere not detected. HPyV6 has not been previously associatedwith any disease, and it is nonetheless possible that theHPyV6 sequences originated from the skin during CSF collection. Whether HPyV6 detection in CSF is associated with patients’ neurological disease, is indicative of contamination with skin microbiota, or reflects increased blood-brain permeability is unknown. Demonstrating a causative role will require further studies. http://dx.doi.org/10.1016/j.jcv.2015.07.156 Abstract No: 1744 Presentation at ESCV 2015: Poster 1 Impact of rapid influenza diagnostic tests on the management of hospitalized patients with suspicion of flu L. Busson1,∗, M. Hallin2, M. De Foor1, B. Mahadeb1, O. Vandenberg3 1 Department of Microbiology, Iris-Lab, Brussels, Belgium 2 Department of Molecular Diagnosis, Iris-Lab, Brussels, Belgium 3 Infectious Diseases Epidemiological Unit, Public Health School, Universite Libre de Bruxelles, Brussels,

MSPrecise: A molecular diagnostic test for multiple sclerosis using next generation sequencing

BackgroundWe have previously demonstrated that cerebrospinal fluid-derived B cells from early relapsing–remitting multiple sclerosis (RRMS) patients that express a VH4 gene accumulate specific replacement mutations. These mutations can be quantified as a score that identifies such patients as having or likely to convert to RRMS. Furthermore, we showed that next generation sequencing is an efficient method for obtaining the sequencing information required by this mutation scoring tool, originally developed using the less clinically viable single-cell Sanger sequencing. ObjectiveTo determine the accuracy of MSPrecise, the diagnostic test that identifies the presence of the RRMS-enriched mutation pattern from patient cerebrospinal fluid B cells. MethodsCerebrospinal fluid cell pellets were obtained from RRMS and other neurological disease (OND) patient cohorts. VH4 gene segments were amplified, sequenced by next generation sequencing and analyzed for mutation score. ResultsThe diagnostic test showed a sensitivity of 75% on the RRMS cohort and a specificity of 88% on the OND cohort. The accuracy of the test in identifying RRMS patients or patients that will develop RRMS is 84%. ConclusionMSPrecise exhibits good performance in identifying patients with RRMS irrespective of time with RRMS.

Intrathecal CD8 T-cells of multiple sclerosis patients recognize lytic Epstein-Barr virus proteins

Background: The association between Epstein-Barr virus (EBV) and multiple sclerosis (MS) may involve intrathecal EBV-specific T-cell responses targeting the virus or indirectly, autoantigens. Objective: Compare the prevalence and fine-specificity of EBV-specific T-cells in the cerebrospinal fluid (CSF) of patients with MS (n = 12), clinically-isolated syndrome (CIS) (n = 17) and other neurological diseases (OND) (n = 13). Methods: Intrathecal EBV-specific T-cell reactivity was assayed using CSF-derived T-cell lines (CSF-TCL) and autologous EBV-transformed B-cells (autoBLCL) as antigen-presenting cells (APC). EBV proteins recognized by autoBLCL-specific CD8 T-cells were identified using human leukocyte antigen class I (HLA-I)-negative monkey cells as artificial APC, co-transfected with 59 different EBV genes and the corresponding patient’s HLA-I alleles that were involved in autoBLCL T-cell reactivity. Reactivity towards the MS-associated autoantigen αB-crystallin (CRYAB) was determined analogously. Results: CSF-TCL from CIS and MS patients had significantly higher frequencies of autoBLCL-reactive CD4 T-cells, compared to the OND patients. CIS patients also had significantly higher autoBLCL-reactive CD8 T cells, which correlated with reactive CD4 T-cell frequencies. AutoBLCL-specific CD8 T-cell responses of four CSF-TCL analyzed in detail were oligoclonal and directed to lytic EBV proteins, but not CRYAB endogenously expressed by autoBLCL. Conclusions: Enhanced intrathecal autoBLCL-specific T-cell reactivity, selectively directed towards lytic EBV proteins in two CSF-TCL, suggested a localized T-cell response to EBV in patients with MS. Our data warrant further characterization of the magnitude and breadth of intrathecal EBV-specific T-cell responses in larger patient cohorts.

MSPrecise: a new diagnostic test for relapsing-remitting multiple sclerosis by cerebrospinal fluid B cell sequencing (HUM3P.267)

Abstract Patients with relapsing-remitting multiple sclerosis (RRMS), or who will convert to RRMS, have cerebrospinal fluid (CSF) B cells that accumulate specific replacement mutations in their rearranged VH4 gene segments as identified by single-cell Sanger sequencing. To harness this observation in a clinical setting, we have developed a diagnostic test to extract and score this genetic information from bulk CSF lymphocytes by next-generation DNA sequencing (NGS). Our current aim is to determine the accuracy of this test on a cohort of patients with varying times of RRMS onset and disease modifying therapies, as well as on a diverse cohort of patients with other neurological diseases (OND) that generally mimic the presentation of MS. Our initial testing on 39 patients has shown an accuracy of 84% in distinguishing RRMS from OND patients. As we continue to expand the size of the cohorts being tested, we will also focus on identifying differences between sub-cohorts of patients at different stages of RRMS or with particular OND diagnoses. With a growing need for a faster and more accurate diagnosis of RRMS, MSPrecise may be a useful addition to the existing diagnostic toolkit that clinicians use to achieve this goal.

Antifibroblast growth factor receptor 3 antibodies identify a subgroup of patients with sensory neuropathy

BackgroundImmunological mechanisms are suspected in sensory neuropathy (SN) occurring with systemic autoimmune diseases and in some idiopathic cases, but so far there are no antibodies (Abs) identifying these neuropathies.MethodsIn the...

Hexosylceramides as intrathecal markers of worsening disability in multiple sclerosis

Background: Sphingolipids are important components of neurons and the myelin sheath whose levels are altered in multiple sclerosis (MS). Objectives: We aimed to determine if cerebrospinal fluid (CSF) sphingolipids can be used as markers of MS disease progression. Methods: Using liquid chromatography tandem mass spectrometry, we analysed sphingolipids in CSF from 134 individuals. The MS group included 65 patients divided into 41 relapsing–remitting MS (RRMS) and 24 progressive MS (ProgMS). In addition, a group of 13 early MS/clinically isolated syndrome (EarlyMS) and two control groups consisting of 38 individuals with other neurological diseases (OND) and 18 OND with signs of inflammation (iOND) were analysed. A follow-up study included 17 additional RRMS patients sampled at two time points 4.7±1.7 years apart. Results: Levels of sphingomyelin (SM)- and hexosylceramide (HexCer)-derived sphingolipids increased in the CSF of patients with MS independently of the fatty acid chain length in RRMS (p<0.05). Levels of palmitic acid (16:0)-containing HexCer (HexCer16:0) increased significantly in ProgMS compared with the OND (p<0.001), iOND (p<0.05) and EarlyMS (p<0.01) groups and correlated with Expanded Disability Status Scale in RRMS in both studies (p=0.048; p=0.027). Conclusion: HexCer16:0 is a promising candidate marker of disease progression in MS, especially in RRMS.

Increased autotaxin activity in multiple sclerosis

Autotaxin (ATX) is an enzyme producing lysophosphatidic acid (LPA) from lysophosphatidyl choline (LPC) and it is up-regulated in inflammatory conditions such as various cancers, arthritis and multiple sclerosis (MS). Numerous studies have shown that the LPA signaling gives rise to angiogenesis, mitosis, cell proliferation and cytokine secretion. On the one hand, an increasing body of evidence suggests that blockade of ATX has anti-inflammatory properties in a variety of diseases. The aim of this study was to measure the enzyme activity of ATX in cerebrospinal fluid (CSF) and serum of patients with MS using an enzymatic photometric method. Twenty definite relapsing remitting MS patients along with 20 patients with other neurological diseases (OND) were recruited. The results showed that ATX activity was significantly higher (p value<0.0001) in MS patients than those patients diagnosed with OND. It is possible that inhibition of the ATX may decrease the rate of MS relapses/progression.

Lack of confirmation of anti-inward rectifying potassium channel 4.1 antibodies as reliable markers of multiple sclerosis

Background: auto-antibodies against the potassium channel inward rectifying potassium channel 4.1 (Kir4.1) have previously been identified in 46% of patients with multiple sclerosis (MS). Objectives: to confirm these findings. Methods: we evaluated the presence of anti-Kir4.1 antibodies by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence in 268 MS patients, 46 patients with other neurological diseases (OND) and 45 healthy controls. Results: anti-Kir4.1 antibodies were found in 7.5% of MS patients, 4.3% of OND patients and 4.4% of healthy controls. Immunofluorescence analysis did not identify any specific staining. Conclusions: we confirmed the presence of anti-Kir4.1 antibodies in MS patients, but at a much lower prevalence than previously reported.