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Inhibition of soluble TNF protects mice against brain inflammation and demyelination in a cuprizone model for multiple sclerosis
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Abstracts
Conclusion: Our results indicate that proteome profile of the urine is different in MS and NMO compared to healthy subjects and urine obtained from NMO and MS also differ. In the next phase, paired urine and plasma samples have been collected from additional patients with anti-AQP4+ NMO, MS and HS for validation and more thorough analysis of the proteome, and quantitative analysis of the individual molecules. Urine may be useful to identify biomarkers for MS and NMO. doi:10.1016/j.jneuroim.2014.08.487
Pathology 128 Structural abnormalities of myelin and axon in murine models of CNS demyelinating disease Yoshio Bando, Shigetaka Yoshida Department of Functional Anatomy and Neuroscience, Asahikawa Medical University, Asahikawa, Japan Multiple sclerosis (MS) is the most common chronic inflammatory demyelinating disease of the CNS and demyelination and axonal damage are responsible for neurological deficits in MS. However, the pathological changes that underlie these morphologies are not fully understood. To address this issue, myelin basic protein-deficient shiverer mice and mice with experimental autoimmune encephalomyelitis (EAE) were examined how the morphological differences of myelin and the axon were induced by dysmyelination and demyelination. Here we used osmium-maceration scanning electron microscopic (SEM) analysis. Osmium-maceration SEM displayed clear variations in the ultrastructural abnormalities of myelin structure and axonal organelles in the white matter of the EAE spinal cord. For example, myelin detachment and excess myelin formation were observed as typical abnormalities of myelin during demyelination in EAE. Importantly, compact myelin was well preserved even though in these situations. SEM images also showed the morphological changes of axonal intracellular organelles during demyelination. Enriched mitochondria and well-developed sER-like structures were observed in the axons of the EAE spinal cord. These observations were also found in the axon of shiverer mice. Taken together, these results indicate that there is a close relationship between myelin and axon morphology during demyelination. doi:10.1016/j.jneuroim.2014.08.488
575 Inhibition of soluble TNF protects mice against brain inflammation and demyelination in a cuprizone model for multiple sclerosis Maria Karamitaa, Chris Barnumb, Ray Tesic, David Szymkowskid, Malu Tanseye, Lesley Probertf a
Laboratory of Molecular Genetics, Hellenic Pasteur Institute, Athens, Greece; bSchool of Medicine, Emory University, Atlanta, GA, United States; c FPRT Bio, WA, United States; dXencor, Monrovia, CA, United States; eSchool of Medicine, Emory, Atlanta, GA, United States; fLaboratory of Molecular Genetics, Hellenic Pasteur Institute, Athens, Greece
Tumor necrosis factor (TNF) mediates chronic inflammatory pathologies including those affecting the central nervous system (CNS), but non-selective TNF inhibitors exacerbate multiple sclerosis (MS) and TNF receptor SF1A is associated with MS, indicating beneficial effects of TNF in CNS pathology. In this study, we compared the effects of soluble (solTNF) and transmembrane TNF (tmTNF) in a mouse model of chronic MS induced by oral administration of the neurotoxin cuprizone (CPZ). In this model disease is compartmentalized behind an intact blood brain barrier (BBB) and monitored by widespread activation and proliferation of microglia, astrocytes and oligodendrocyte precursor cells (OPC) after 2 weeks of CPZ and progressive demyelination and neurodegeneration, most obvious in the corpus callosum, with maximum lesions being observed at 5 weeks of administration. Withdrawal of toxin from the diet allows almost complete remyelination. In this study, C57Bl/6 mice were fed ad libitum with CPZ-supplemented diet and from the third week were treated twice-weekly s.c. with XPro1595 (10 mg/kg), a PEGylated 27 kDa dominant-negative analogue of human TNF that selectively blocks solTNF, or etanercept which non-specifically blocks tmTNF and solTNF. Mice were sacrificed at week 5 and brains were processed for neuropathological analysis and measurement of brain levels of dominant-negative human TNF (XPro1595). XPro1595-treated mice showed detectable levels of XPro1595 in the brain, markedly reduced demyelination and microglial activation and proliferation, coupled with increased numbers OPC in lesions compared to controls and etanercept-treated mice. Our results show that in the CPZ-induced model of chronic MS, selective inhibition of solTNF by peripheral administration of XPro1595 prevents neuroinflammation and demyelination, at least partly by increasing the number of OPC in the lesion. These therapeutic effects of XPro1595, coupled with its ability to cross the BBB, have important implications for the treatment of chronic CNS inflammation associated with neurodegeneration. doi:10.1016/j.jneuroim.2014.08.489
103 Brain positron emission tomography scanning can be used to image pathological determinants of progressive multiple sclerosis Laura Airasa, Eero Rissanenb, Maria Gardbergc, Marcus Sucksdorffb, Jouni Tuiskua, Juha Rinneb a
Division of Clinical Neurosciences, Turku University Hospital, Turku, Finland; bTurku PET Centre, Turku University Hospital, Turku, Finland; c Department of Pathology, Turku University Hospital, Turku, Finland Background: In secondary progressive multiple sclerosis (SPMS), the neuropathology is characterized by diffuse inflammation, which is detected also outside the demyelinating lesions and is associated with microglial activation. Objectives: The aim of this study was to evaluate, whether in vivo positron emission tomography (PET) imaging could be reliably used to obtain information about the underlying neuropathology in patients with SPMS. Methods: Ten SPMS patients and eight controls were studied using the radioligand [11C]PK11195 binding to the 18-kDa translocator protein (TSPO) expressed on activated microglial cells. T1-hypointense lesions were classified as [11C]PK11195-positive, chronic active lesions if there was increased [11C]PK11195-DVR covering more than half of the lesion's circumference (in all axial slices) compared to the adjacent or contralateral NAWM. To evaluate the binding pattern of TSPO-ligands in more detail, autoradiography experiments using postmortem human
Abstracts
Conclusion: Our results indicate that proteome profile of the urine is different in MS and NMO compared to healthy subjects and urine obtained from NMO and MS also differ. In the next phase, paired urine and plasma samples have been collected from additional patients with anti-AQP4+ NMO, MS and HS for validation and more thorough analysis of the proteome, and quantitative analysis of the individual molecules. Urine may be useful to identify biomarkers for MS and NMO. doi:10.1016/j.jneuroim.2014.08.487
Pathology 128 Structural abnormalities of myelin and axon in murine models of CNS demyelinating disease Yoshio Bando, Shigetaka Yoshida Department of Functional Anatomy and Neuroscience, Asahikawa Medical University, Asahikawa, Japan Multiple sclerosis (MS) is the most common chronic inflammatory demyelinating disease of the CNS and demyelination and axonal damage are responsible for neurological deficits in MS. However, the pathological changes that underlie these morphologies are not fully understood. To address this issue, myelin basic protein-deficient shiverer mice and mice with experimental autoimmune encephalomyelitis (EAE) were examined how the morphological differences of myelin and the axon were induced by dysmyelination and demyelination. Here we used osmium-maceration scanning electron microscopic (SEM) analysis. Osmium-maceration SEM displayed clear variations in the ultrastructural abnormalities of myelin structure and axonal organelles in the white matter of the EAE spinal cord. For example, myelin detachment and excess myelin formation were observed as typical abnormalities of myelin during demyelination in EAE. Importantly, compact myelin was well preserved even though in these situations. SEM images also showed the morphological changes of axonal intracellular organelles during demyelination. Enriched mitochondria and well-developed sER-like structures were observed in the axons of the EAE spinal cord. These observations were also found in the axon of shiverer mice. Taken together, these results indicate that there is a close relationship between myelin and axon morphology during demyelination. doi:10.1016/j.jneuroim.2014.08.488
575 Inhibition of soluble TNF protects mice against brain inflammation and demyelination in a cuprizone model for multiple sclerosis Maria Karamitaa, Chris Barnumb, Ray Tesic, David Szymkowskid, Malu Tanseye, Lesley Probertf a
Laboratory of Molecular Genetics, Hellenic Pasteur Institute, Athens, Greece; bSchool of Medicine, Emory University, Atlanta, GA, United States; c FPRT Bio, WA, United States; dXencor, Monrovia, CA, United States; eSchool of Medicine, Emory, Atlanta, GA, United States; fLaboratory of Molecular Genetics, Hellenic Pasteur Institute, Athens, Greece
Tumor necrosis factor (TNF) mediates chronic inflammatory pathologies including those affecting the central nervous system (CNS), but non-selective TNF inhibitors exacerbate multiple sclerosis (MS) and TNF receptor SF1A is associated with MS, indicating beneficial effects of TNF in CNS pathology. In this study, we compared the effects of soluble (solTNF) and transmembrane TNF (tmTNF) in a mouse model of chronic MS induced by oral administration of the neurotoxin cuprizone (CPZ). In this model disease is compartmentalized behind an intact blood brain barrier (BBB) and monitored by widespread activation and proliferation of microglia, astrocytes and oligodendrocyte precursor cells (OPC) after 2 weeks of CPZ and progressive demyelination and neurodegeneration, most obvious in the corpus callosum, with maximum lesions being observed at 5 weeks of administration. Withdrawal of toxin from the diet allows almost complete remyelination. In this study, C57Bl/6 mice were fed ad libitum with CPZ-supplemented diet and from the third week were treated twice-weekly s.c. with XPro1595 (10 mg/kg), a PEGylated 27 kDa dominant-negative analogue of human TNF that selectively blocks solTNF, or etanercept which non-specifically blocks tmTNF and solTNF. Mice were sacrificed at week 5 and brains were processed for neuropathological analysis and measurement of brain levels of dominant-negative human TNF (XPro1595). XPro1595-treated mice showed detectable levels of XPro1595 in the brain, markedly reduced demyelination and microglial activation and proliferation, coupled with increased numbers OPC in lesions compared to controls and etanercept-treated mice. Our results show that in the CPZ-induced model of chronic MS, selective inhibition of solTNF by peripheral administration of XPro1595 prevents neuroinflammation and demyelination, at least partly by increasing the number of OPC in the lesion. These therapeutic effects of XPro1595, coupled with its ability to cross the BBB, have important implications for the treatment of chronic CNS inflammation associated with neurodegeneration. doi:10.1016/j.jneuroim.2014.08.489
103 Brain positron emission tomography scanning can be used to image pathological determinants of progressive multiple sclerosis Laura Airasa, Eero Rissanenb, Maria Gardbergc, Marcus Sucksdorffb, Jouni Tuiskua, Juha Rinneb a
Division of Clinical Neurosciences, Turku University Hospital, Turku, Finland; bTurku PET Centre, Turku University Hospital, Turku, Finland; c Department of Pathology, Turku University Hospital, Turku, Finland Background: In secondary progressive multiple sclerosis (SPMS), the neuropathology is characterized by diffuse inflammation, which is detected also outside the demyelinating lesions and is associated with microglial activation. Objectives: The aim of this study was to evaluate, whether in vivo positron emission tomography (PET) imaging could be reliably used to obtain information about the underlying neuropathology in patients with SPMS. Methods: Ten SPMS patients and eight controls were studied using the radioligand [11C]PK11195 binding to the 18-kDa translocator protein (TSPO) expressed on activated microglial cells. T1-hypointense lesions were classified as [11C]PK11195-positive, chronic active lesions if there was increased [11C]PK11195-DVR covering more than half of the lesion's circumference (in all axial slices) compared to the adjacent or contralateral NAWM. To evaluate the binding pattern of TSPO-ligands in more detail, autoradiography experiments using postmortem human