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No correlation between Kir 4.1 autoimmunity and multiple sclerosis
Abstracts
Multiple sclerosis (MS) is one of the leading causes of disability among young adults. In approximately 80–85% of MS patients, the disease follows a relapsing–remitting course (RRMS). Over time, the majority of these patients will enter a progressive phase (secondary progressive MS– SPMS). The remaining 15–20% of MS patients follow an essentially progressive course from the beginning and they are referred to as primary progressive MS (PPMS). MS disease course is highly unpredictable and biomarkers that may help in the distinction between different activity phases of MS are unfortunately lacking. In the present study, we aimed to characterize the different clinical forms and activity phases of MS by means of transcriptomics and immunophenotyping approaches. A total of 47 untreated MS patients and 19 healthy controls were included in the study. The MS group included 10 patients with early RRMS (in the first 5 years after disease onset), 10 patients with SPMS, 13 patients with PPMS, 10 patients with benign disease course (defined by EDSS ≤3 after 10 years from disease onset), and 4 patients with aggressive disease courses (defined by the presence over the last year of 2 or more relapses and increase of at least one point in the EDSS). The gene expression profiling was determined in peripheral blood mononuclear cells (PBMC) from MS patients and healthy controls using microarrays (Affymetrix HGU219 chip). Immunophenotyping of PBMC was performed by cell surface staining for monocytes, NK, B, and T cells and subsequent flow cytometry analysis. PBMC from MS patients with benign disease course were characterized by an over-expression of TNF compared with cells from patients with other clinical forms and activity phases of the disease and healthy controls. A large number of TNF-induced genes were also significantly up-regulated in PBMC from patients with benign MS, which included IL1B, CXCL2, CXCL3, CCL3, IER3, NFKBIA, and TNFAIP3 among others. Microarray findings were validated by real time PCR. Immunophenotyping of PBMC revealed that patients with benign forms of MS showed a significant increase in the frequency of CD16/56+ NK cells compared to other MS groups and healthy controls. The TNF gene expression signature and the increased frequency of NK cells in PBMC may be used as biomarkers to discriminate between different activity phases of MS. doi:10.1016/j.jneuroim.2014.08.053
551 No correlation between Kir 4.1 autoimmunity and multiple sclerosis Justyna Maria Czarna Bahl Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, Copenhagen, Denmark Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease, manifested as lesions in the white matter of the central nervous system. Correspondingly, MS results in either temporary or permanent neurological deficits. Progress in the treatment of MS has increased the focus on quick diagnosis and risk of progression from clinically isolated syndrome, e.g. optic neuritis (ON), to manifest MS and on monitoring the effect of therapeutic intervention. Recently, Srivastava et al. have shown that serum from patients with MS reacted against an extracellular loop of the rectifying potassium channel 4.1 (Kir4.1). In this study we have analysed whether autoantibodies against a Kir4.1 synthetic peptide could discriminate 336 consecutively referred patients from the Department of Neurology with MS (n = 70), ON (n = 188) or neuromyelitis optica (NMO, n = 8) from controls with other neurological symptoms (n = 70) as well as healthy controls (n = 100). Moreover, we correlated the Kir4.1 and Aquaporin 4 autoimmunity in patients with NMO and those with ON as clinically isolated syndrome. We used an ELISA set-up with a synthetic peptide of
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the extracellular loop (amino acids 83–120) of Kir4.1 coated onto the plates. Results: Low titres against kir4.1 were seen in all groups. The titers in diseased patients were the same as in the healthy controls. Conclusions: We have analysed the Kir4.1 reactivity in a large group of well-defined patients and controls. We do not find any difference in Kir4.1 peptide response in serum between patients and controls. Results: Low titres against kir4.1 were seen in all groups. The titers in diseased patients were the same as in the healthy controls. Conclusion: We have analysed the Kir4.1 reactivity in a large group of well-defined patients and controls. We do not find any difference in Kir4.1 peptide response in serum between patients and controls. doi:10.1016/j.jneuroim.2014.08.054
645 Cerebrospinal fluid light neurofilaments as a therapeutic marker of response in modifying therapies disease in multiple sclerosis Fabian Gomez, Carlos Oehninger Hospital Clinic, Faculty of Medicine, Uruguay Introduction: During the degenerative processes in the central nervous system in multiple sclerosis destruction of the neuronal cytoskeleton pathway mediated by calpains with neurofilament release into the extracellular space occurs, so its determination in the cerebrospinal fluid is seen as a key biomarker predicting the load of axonal injury and the effectiveness of immunomodulatory therapies to long-term disability. Neurofilament light are structural components of the neuronal cytoskeleton that are released into the extracellular space in inflammatory and degenerative processes of multiple sclerosis during microglial activation and that would be key biomarkers of axonal injury and long term disability, essential to measure the effectiveness of therapies immunomodulatory. Aims and objectives: The overall objective of this study is the determination of neurofilament light levels in CSF as a marker of axonal damage and therapeutic response in patients which treatment is initiated with interferon beta 1a, 1b, glatiramer acetate, fingolimod and natalizumab. Study design: A cohort study of a group of 60 patients diagnosed with multiple sclerosis will be held according to the revised McDonald criteria in 2010 with a lower EDSS of 3.0 starting treatment with immunomodulatory therapies, to which they underwent clinical assessment with EDSS, Multiple Sclerosis Functional Composite (MSFC9 scales), imaging with MRI and determination liquoral with neurofilament light (NFL) in CSF at baseline and then every six months for two years. Methodology: All patients in the initial stage of the study will be evaluated clinically by routine neurological examination, and functionality are determined with EDSS and MSFC. Subsequently, a lumbar puncture was carried out with removal of 5–10 ml CSF during clinical thrust, and will be booked under refrigeration at –80° for the determination of the neurofilament-light available ELISA kit (Diagnostics AB Uman). In the evolution under immunomodulatory treatment neurofilament will be measured at 12, 18 and 24 months. Expected results: Regarding the expected results we determine neurofilament CSF at the beginning of the study will be dependent on the timing and aggressiveness of the disease, and their levels will be minor in the monthly and annual checks primarily to aggressive therapies such as natalizumab and fingolimod, being of lesser importance for first-line drugs such as interferons and glatiramer acetate. This work once again affirm previous findings in other cohorts studied for prognostic value of CSF neurofilament as a biomarker of therapeutic response to drugs of first line and second line on axonal damage and long-term disability.
Multiple sclerosis (MS) is one of the leading causes of disability among young adults. In approximately 80–85% of MS patients, the disease follows a relapsing–remitting course (RRMS). Over time, the majority of these patients will enter a progressive phase (secondary progressive MS– SPMS). The remaining 15–20% of MS patients follow an essentially progressive course from the beginning and they are referred to as primary progressive MS (PPMS). MS disease course is highly unpredictable and biomarkers that may help in the distinction between different activity phases of MS are unfortunately lacking. In the present study, we aimed to characterize the different clinical forms and activity phases of MS by means of transcriptomics and immunophenotyping approaches. A total of 47 untreated MS patients and 19 healthy controls were included in the study. The MS group included 10 patients with early RRMS (in the first 5 years after disease onset), 10 patients with SPMS, 13 patients with PPMS, 10 patients with benign disease course (defined by EDSS ≤3 after 10 years from disease onset), and 4 patients with aggressive disease courses (defined by the presence over the last year of 2 or more relapses and increase of at least one point in the EDSS). The gene expression profiling was determined in peripheral blood mononuclear cells (PBMC) from MS patients and healthy controls using microarrays (Affymetrix HGU219 chip). Immunophenotyping of PBMC was performed by cell surface staining for monocytes, NK, B, and T cells and subsequent flow cytometry analysis. PBMC from MS patients with benign disease course were characterized by an over-expression of TNF compared with cells from patients with other clinical forms and activity phases of the disease and healthy controls. A large number of TNF-induced genes were also significantly up-regulated in PBMC from patients with benign MS, which included IL1B, CXCL2, CXCL3, CCL3, IER3, NFKBIA, and TNFAIP3 among others. Microarray findings were validated by real time PCR. Immunophenotyping of PBMC revealed that patients with benign forms of MS showed a significant increase in the frequency of CD16/56+ NK cells compared to other MS groups and healthy controls. The TNF gene expression signature and the increased frequency of NK cells in PBMC may be used as biomarkers to discriminate between different activity phases of MS. doi:10.1016/j.jneuroim.2014.08.053
551 No correlation between Kir 4.1 autoimmunity and multiple sclerosis Justyna Maria Czarna Bahl Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, Copenhagen, Denmark Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease, manifested as lesions in the white matter of the central nervous system. Correspondingly, MS results in either temporary or permanent neurological deficits. Progress in the treatment of MS has increased the focus on quick diagnosis and risk of progression from clinically isolated syndrome, e.g. optic neuritis (ON), to manifest MS and on monitoring the effect of therapeutic intervention. Recently, Srivastava et al. have shown that serum from patients with MS reacted against an extracellular loop of the rectifying potassium channel 4.1 (Kir4.1). In this study we have analysed whether autoantibodies against a Kir4.1 synthetic peptide could discriminate 336 consecutively referred patients from the Department of Neurology with MS (n = 70), ON (n = 188) or neuromyelitis optica (NMO, n = 8) from controls with other neurological symptoms (n = 70) as well as healthy controls (n = 100). Moreover, we correlated the Kir4.1 and Aquaporin 4 autoimmunity in patients with NMO and those with ON as clinically isolated syndrome. We used an ELISA set-up with a synthetic peptide of
19
the extracellular loop (amino acids 83–120) of Kir4.1 coated onto the plates. Results: Low titres against kir4.1 were seen in all groups. The titers in diseased patients were the same as in the healthy controls. Conclusions: We have analysed the Kir4.1 reactivity in a large group of well-defined patients and controls. We do not find any difference in Kir4.1 peptide response in serum between patients and controls. Results: Low titres against kir4.1 were seen in all groups. The titers in diseased patients were the same as in the healthy controls. Conclusion: We have analysed the Kir4.1 reactivity in a large group of well-defined patients and controls. We do not find any difference in Kir4.1 peptide response in serum between patients and controls. doi:10.1016/j.jneuroim.2014.08.054
645 Cerebrospinal fluid light neurofilaments as a therapeutic marker of response in modifying therapies disease in multiple sclerosis Fabian Gomez, Carlos Oehninger Hospital Clinic, Faculty of Medicine, Uruguay Introduction: During the degenerative processes in the central nervous system in multiple sclerosis destruction of the neuronal cytoskeleton pathway mediated by calpains with neurofilament release into the extracellular space occurs, so its determination in the cerebrospinal fluid is seen as a key biomarker predicting the load of axonal injury and the effectiveness of immunomodulatory therapies to long-term disability. Neurofilament light are structural components of the neuronal cytoskeleton that are released into the extracellular space in inflammatory and degenerative processes of multiple sclerosis during microglial activation and that would be key biomarkers of axonal injury and long term disability, essential to measure the effectiveness of therapies immunomodulatory. Aims and objectives: The overall objective of this study is the determination of neurofilament light levels in CSF as a marker of axonal damage and therapeutic response in patients which treatment is initiated with interferon beta 1a, 1b, glatiramer acetate, fingolimod and natalizumab. Study design: A cohort study of a group of 60 patients diagnosed with multiple sclerosis will be held according to the revised McDonald criteria in 2010 with a lower EDSS of 3.0 starting treatment with immunomodulatory therapies, to which they underwent clinical assessment with EDSS, Multiple Sclerosis Functional Composite (MSFC9 scales), imaging with MRI and determination liquoral with neurofilament light (NFL) in CSF at baseline and then every six months for two years. Methodology: All patients in the initial stage of the study will be evaluated clinically by routine neurological examination, and functionality are determined with EDSS and MSFC. Subsequently, a lumbar puncture was carried out with removal of 5–10 ml CSF during clinical thrust, and will be booked under refrigeration at –80° for the determination of the neurofilament-light available ELISA kit (Diagnostics AB Uman). In the evolution under immunomodulatory treatment neurofilament will be measured at 12, 18 and 24 months. Expected results: Regarding the expected results we determine neurofilament CSF at the beginning of the study will be dependent on the timing and aggressiveness of the disease, and their levels will be minor in the monthly and annual checks primarily to aggressive therapies such as natalizumab and fingolimod, being of lesser importance for first-line drugs such as interferons and glatiramer acetate. This work once again affirm previous findings in other cohorts studied for prognostic value of CSF neurofilament as a biomarker of therapeutic response to drugs of first line and second line on axonal damage and long-term disability.