- Email: [email protected]
CNS tissue response to inflammation
Abstracts
Results: In rat strains we identified Atg7 as an EAE associated gene, and further showed that higher Atg7 expression correlates with less severe EAE. The mouse strain with conditional Atg7 deletion in myeloid shows aggravated EAE. The impact on EAE chronicity is associated with elevated Neutrophil proportions in EAE CNS at later stages. In PBMC and sorted immune cells from MS patients and controls we detected altered mRNA expression patterns. Furthermore, we observed an association of Atg7 to MS in a Swedish population. Conclusions: In conclusion, we demonstrate a regulation of Atg7 expression in MS and EAE immune cells and an impact on EAE disease phenotypes. To elucidate the molecular impact of Atg7 and autophagy in neuroinflammatory disease further functional studies are required. Based on our findings our primary interests are in Neutrophils and other myeloid cells targeting autophagy, regulatory- and executive properties.
doi:10.1016/j.jneuroim.2014.08.366
327 CNS tissue response to inflammation Sabina Berl, Ari Waisman Molecular Medicine, Johannes Gutenberg University, Mainz, Germany A well known inflammatory, autoimmune disease of the central nervous system (CNS) is Multiple sclerosis (MS), where cells of the immune system invade the CNS, causing demyelination and neurodegeneration and leading to paralysis. The most common animal model of MS is experimental autoimmune encephalomyelitis (EAE), which can be induced by immunization of mice with myelin antigens. Not much is known about the response of neurons to the inflammatory triggers, although damage to them is the main initiator for paralysis in MS patients and EAE, because of the difficulty to separate and analyze them for gene expression on single cell level. The aim of this project is to isolate and further analyze neuronal mRNA expression level from adult mice. Isolated neuronal mRNA of naïve mice and mice with EAE will be sequenced by Next Generation Sequencing (NGS) to better understand how neurons respond to damage induced by inflammation. This study has implications for MS research and other inflammatory disease of the CNS. We immunoprecipitate mRNA with a modified Ribosomal Protein L22 including a HA-tag, using the RiboTag mice. After crossing the RiboTag mice to the neuron-specific NFH-Cre mice, HA-tagged ribosomes will be specifically expressed in neurons giving the possibility to immunoprecipitate only neuronal RNA. Isolated RNA from immunoprecipitated material of the RiboTag mice will be further analyzed by deep sequencing using NGS technology in collaboration with sequencing core facility of Johannes-GutenbergUniversity of Mainz. The final goal of this study is to analyze gene expression pattern of neurons that are attacked by pro-inflammatory cells and mediators during EAE. Understanding how neurons respond during autoimmune disease should allow to design better therapies that will protect these cells under such pathogenic conditions. This work has implications not only for patients of Multiple sclerosis, but also for other neurological diseases, including Alzheimer's disease and stroke.
635 Effect of Natalizumab treatment on circulating CD4 + CD62L+ T-cells in Multiple Sclerosis patients Antonio Bertolottoa, Fabiana Marnettoa, Marzia Caldanoa, Alessandra Lugaresib, Michela Spadaroa a AOU San Luigi Gonzaga, Clinical Neurobiology Unit, Regional Referring Multiple Sclerosis Center (CRESM), Orbassano, Turin, Italy; bDept. Neuroscience, Imaging and Clinical Sciences, University “G. d'Annunzio”, Chieti, Italy
L-selectin or CD62L is an adhesion molecule involved in rolling and transmigration of leukocyte cells. Recently, Schwab and colleagues showed a correlation between a low expression of CD62L on CD4+ T-cells and the development of progressive multifocal leukoencephalopathy (PML) in Natalizumab (NTZ)-treated Multiple Sclerosis (MS) patients. To verify the effect of NTZ treatment on the expression of CD62L on CD4+ T cells and the possible correlation with PML, patients were recruited by CRESM, San Luigi, University Hospital. We conducted a flow cytometry analysis on cryopreserved, viable peripheral blood mononuclear cells from 23 healthy donors (HD) and 171 patients with MS: 100 had been treated with NTZ (1–86 months), 52 received other treatments (26 with Interferon beta, IFNb; 26 with Glatiramer acetate, GA) and 19 were untreated (T0). We also analyzed two PML cases in a NTZ-treated patient from the Multiple Sclerosis Center of the University of Chieti. The percentage of L-selectin on CD4+ T cells was significantly lower in patients treated with NTZ (33.70% ± 12.9; mean ± SD) when compared to patients treated with GA (47.03% ± 14.7; p = 0.0002), IFNb (42.95% ± 18.9; p = 0.0477), T0 (43.5% ± 10.8; p = 0.0032), or HD (46.75% ± 9.8; p b 0.0001). Interestingly, we found two outliers for the expression of CD62L+ on CD4+ Tcells in GA-treated group and in T0 patients. In the NTZ-treated group four patients had values lower than the calculated risk threshold 11.05% (mean - 2x SD of IFNb + GA group). The effect of NTZ was already evident during the first year of treatment, and persisted in the following years (p = 0.0078, compared to IFNb + GA group). In the six high risk patients two CD62L serial evaluations were performed, during NTZ treatment and two or four months after withdrawal: during treatment 3 out of 6 had low values of CD4+CD62L+ which increased to normal values after discontinuation. We also had the opportunity to analyze two patients in the acute PML/IRIS phase, 4 months after NTZ withdrawal and under steroid treatment, CD62L expression were 30.58% and 39.39%. NTZ treatment decreases the expression of CD62L on circulating CD4+ T cells. This effect is already evident in the first year of treatment and disappears after withdrawal of treatment. The study of the effect of NTZ on CD4+CD62L+ cells can be a valuable tool to personalize NTZ therapy.
doi:10.1016/j.jneuroim.2014.08.368
307 Neuropathological evaluation of four Danish cases of CLIPPERS: Evidence of generalized neuroinflammation but no pre-lymphoma Morten Blaabjerga, Bjarne Winther Kristensenb, Daniel Kondziellac, Bjørg Morell Kerrn-jespersenc, Mette Lindelofd, Zsolt Illesa a
Department of Neurology, Odense University Hospital, Odense, Denmark; Department of Pathology, Odense University Hospital, Odense, Denmark; c Department of Neurology, Copenhagen University Hospital, Copenhagen, Denmark; dDepartment of Neurology, Herlev Hospital, Herlev, Denmark b
doi:10.1016/j.jneuroim.2014.08.367
137
Results: In rat strains we identified Atg7 as an EAE associated gene, and further showed that higher Atg7 expression correlates with less severe EAE. The mouse strain with conditional Atg7 deletion in myeloid shows aggravated EAE. The impact on EAE chronicity is associated with elevated Neutrophil proportions in EAE CNS at later stages. In PBMC and sorted immune cells from MS patients and controls we detected altered mRNA expression patterns. Furthermore, we observed an association of Atg7 to MS in a Swedish population. Conclusions: In conclusion, we demonstrate a regulation of Atg7 expression in MS and EAE immune cells and an impact on EAE disease phenotypes. To elucidate the molecular impact of Atg7 and autophagy in neuroinflammatory disease further functional studies are required. Based on our findings our primary interests are in Neutrophils and other myeloid cells targeting autophagy, regulatory- and executive properties.
doi:10.1016/j.jneuroim.2014.08.366
327 CNS tissue response to inflammation Sabina Berl, Ari Waisman Molecular Medicine, Johannes Gutenberg University, Mainz, Germany A well known inflammatory, autoimmune disease of the central nervous system (CNS) is Multiple sclerosis (MS), where cells of the immune system invade the CNS, causing demyelination and neurodegeneration and leading to paralysis. The most common animal model of MS is experimental autoimmune encephalomyelitis (EAE), which can be induced by immunization of mice with myelin antigens. Not much is known about the response of neurons to the inflammatory triggers, although damage to them is the main initiator for paralysis in MS patients and EAE, because of the difficulty to separate and analyze them for gene expression on single cell level. The aim of this project is to isolate and further analyze neuronal mRNA expression level from adult mice. Isolated neuronal mRNA of naïve mice and mice with EAE will be sequenced by Next Generation Sequencing (NGS) to better understand how neurons respond to damage induced by inflammation. This study has implications for MS research and other inflammatory disease of the CNS. We immunoprecipitate mRNA with a modified Ribosomal Protein L22 including a HA-tag, using the RiboTag mice. After crossing the RiboTag mice to the neuron-specific NFH-Cre mice, HA-tagged ribosomes will be specifically expressed in neurons giving the possibility to immunoprecipitate only neuronal RNA. Isolated RNA from immunoprecipitated material of the RiboTag mice will be further analyzed by deep sequencing using NGS technology in collaboration with sequencing core facility of Johannes-GutenbergUniversity of Mainz. The final goal of this study is to analyze gene expression pattern of neurons that are attacked by pro-inflammatory cells and mediators during EAE. Understanding how neurons respond during autoimmune disease should allow to design better therapies that will protect these cells under such pathogenic conditions. This work has implications not only for patients of Multiple sclerosis, but also for other neurological diseases, including Alzheimer's disease and stroke.
635 Effect of Natalizumab treatment on circulating CD4 + CD62L+ T-cells in Multiple Sclerosis patients Antonio Bertolottoa, Fabiana Marnettoa, Marzia Caldanoa, Alessandra Lugaresib, Michela Spadaroa a AOU San Luigi Gonzaga, Clinical Neurobiology Unit, Regional Referring Multiple Sclerosis Center (CRESM), Orbassano, Turin, Italy; bDept. Neuroscience, Imaging and Clinical Sciences, University “G. d'Annunzio”, Chieti, Italy
L-selectin or CD62L is an adhesion molecule involved in rolling and transmigration of leukocyte cells. Recently, Schwab and colleagues showed a correlation between a low expression of CD62L on CD4+ T-cells and the development of progressive multifocal leukoencephalopathy (PML) in Natalizumab (NTZ)-treated Multiple Sclerosis (MS) patients. To verify the effect of NTZ treatment on the expression of CD62L on CD4+ T cells and the possible correlation with PML, patients were recruited by CRESM, San Luigi, University Hospital. We conducted a flow cytometry analysis on cryopreserved, viable peripheral blood mononuclear cells from 23 healthy donors (HD) and 171 patients with MS: 100 had been treated with NTZ (1–86 months), 52 received other treatments (26 with Interferon beta, IFNb; 26 with Glatiramer acetate, GA) and 19 were untreated (T0). We also analyzed two PML cases in a NTZ-treated patient from the Multiple Sclerosis Center of the University of Chieti. The percentage of L-selectin on CD4+ T cells was significantly lower in patients treated with NTZ (33.70% ± 12.9; mean ± SD) when compared to patients treated with GA (47.03% ± 14.7; p = 0.0002), IFNb (42.95% ± 18.9; p = 0.0477), T0 (43.5% ± 10.8; p = 0.0032), or HD (46.75% ± 9.8; p b 0.0001). Interestingly, we found two outliers for the expression of CD62L+ on CD4+ Tcells in GA-treated group and in T0 patients. In the NTZ-treated group four patients had values lower than the calculated risk threshold 11.05% (mean - 2x SD of IFNb + GA group). The effect of NTZ was already evident during the first year of treatment, and persisted in the following years (p = 0.0078, compared to IFNb + GA group). In the six high risk patients two CD62L serial evaluations were performed, during NTZ treatment and two or four months after withdrawal: during treatment 3 out of 6 had low values of CD4+CD62L+ which increased to normal values after discontinuation. We also had the opportunity to analyze two patients in the acute PML/IRIS phase, 4 months after NTZ withdrawal and under steroid treatment, CD62L expression were 30.58% and 39.39%. NTZ treatment decreases the expression of CD62L on circulating CD4+ T cells. This effect is already evident in the first year of treatment and disappears after withdrawal of treatment. The study of the effect of NTZ on CD4+CD62L+ cells can be a valuable tool to personalize NTZ therapy.
doi:10.1016/j.jneuroim.2014.08.368
307 Neuropathological evaluation of four Danish cases of CLIPPERS: Evidence of generalized neuroinflammation but no pre-lymphoma Morten Blaabjerga, Bjarne Winther Kristensenb, Daniel Kondziellac, Bjørg Morell Kerrn-jespersenc, Mette Lindelofd, Zsolt Illesa a
Department of Neurology, Odense University Hospital, Odense, Denmark; Department of Pathology, Odense University Hospital, Odense, Denmark; c Department of Neurology, Copenhagen University Hospital, Copenhagen, Denmark; dDepartment of Neurology, Herlev Hospital, Herlev, Denmark b
doi:10.1016/j.jneuroim.2014.08.367
137