- Email: [email protected]
Evaluating TNF-alpha as a possible link in Alzheimer's disease and rheumatoid arthritis
Sunday, July 14, 2013: Poster Presentations: P1 were found in 5xFAD versus non-Tg mice. Conclusions: Preliminary results suggest there is immune cell activation and neuroinflammation in the 5xFAD by 4 months of age leading to increased levels of immune cells at 6 months of age. Studies are now in progress to determine the source (central versus peripheral) of immune cell activation and extent to which chronic systemic inflammation promotes peripheral immune cell traffic to the CNS.
P1-023
NEUROINFLAMMATION IN HAPPSL TRANSGENIC MICE
Tina L€ offler1, Daniel Havas1, Stefanie Flunkert1, Nicole Taub1, Manfred Windisch2, Birgit Hutter-Paier1, 1QPS Austria, Grambach, Austria; 2QPS, Grambach, Austria. Contact e-mail: rigit. [email protected] Background: Deposition of neurotoxic amyloid beta peptides and the formation of neurofibrillary tangles represent the major pathological hallmarks of Alzheimer’s disease (AD). A rising body of evidence suggests neuroinflammation to be a third decisive component that actively contributes to disease progression and severity. Reactive astrocytes and microglia surrounding senile plaques play a pivotal role in the attempt to fight toxic depositions in AD brain, during which they can act as cytotoxic effector cells by releasing substances such as reactive oxygen species (ROS) and cytokines. These processes, oxidative stress and neuroinflammation, nourish the vicious circle of AD pathology and are thus seen as potential therapeutic targets. Methods: Cortical and hippocampal tissue of 6, 9 and 12 months old hAPP SL transgenic and non-transgenic mice was analyzed for soluble and insoluble Ab1-38, 1-40 and 1-42 with the MSDÒ 96-well MULTI-SPOTÒ 4G8 Ab Triplex Assay (Mesoscale Discovery). Brain tissue was histologically analyzed for astrocytosis and activated microglia by immunofluorescent staining with GFAP-IHC (Dako, Z0334) and CD11b-IHC (Serotec, MCA711) antibody, respectively. Furthermore, tissue of 4, 6, 9 and 12 months old animals was analyzed for oxidative stress by using the TBARS-assay. Results: Starting at an age of 6 months hAPP SL transgenic mice exhibit significantly enhanced Ab1-42 levels in the cortex and hippocampus followed by a significant increase of Ab1-38 and 1-40 at an age of 9 months. Simultaneously, neuroinflammation comprising microgliosis and astrocytosis increases in the cortex. Microgliosis also increases in the hippocampus and corpus callosum. Analysis of oxidative stress in 9 and 12 months old hAPP SL transgenic mice reveals a significant increase in the cortex and hippocampus compared to non-transgenic littermates and correlates with the higher incidence of glial inflammatory response. Conclusions: Next to a severe amyloid pathology, hAPP SL transgenic mice also develop neuroinflammation and oxidative stress at later time points, displaying a broad spectrum of AD pathology. These mice represent therefore a valid tool, not only to screen developmental compounds interfering with amyloid generation and plaque formation, but also anti-inflammatory and/ or immune-modulatory agents.
P1-024
IMMUNOLOGICAL DYSFUNCTION AND MCILIKE BEHAVIOR IN THE 3XTGAD MICE
Monica Marchese1, David Cowan1, Khalil Karimi1, Vanessa Ashthorpe1, Elizabeth Head2, Donglai Ma1, Hui Zhao1, Paulina Davis3, Minesh Kapadia1, Boris Sakic1, 1McMaster University, Hamilton, Ontario, Canada; 2Sanders-Brown Center on Aging, University of Kentucky, Lexington, Kentucky, United States; 3University of Kentucky, Lexington, Kentucky, United States. Contact e-mail: [email protected] Background: Although immune system activation has been observed in patients with Alzheimer’s disease (AD), it remains unclear whether inflammatory and/or autoimmune manifestations are epiphenomena, consequence, or cause of brain degeneration. We presently examine whether immunological changes accompany the progress of AD-like disease in well-established 3xTgAD murine model. Methods: We compared the immunological and behavioral profiles of triple transgenic mice (3xTgAD) and wild type (WT) controls, from 1.5 to 12 months of age. Mice underwent a broad battery of tests to assess longitudinal changes in behavioral performance.
P161
FACScan analysis and immunoenzymatic assays were employed to quantify inflammatory and autoimmune markers in serum and spleen. Results: Prior to the development of advanced AD pathology, 3xTgAD mice developed MCI-like manifestations, characterized by anxiety-related behaviors, changes in olfactory function and impaired flexibility in learning/memory tasks (2.5 months). Several manifestations of systemic inflammatory / autoimmune disease were observed at later age (12 months). They included splenomegaly, elevated serum levels of anti-nuclear and anti-dsDNA antibodies, as well as a reduced number of T splenocytes. Changes in the immune system occurred in parallel with generalized learning/memory and olfaction deficits from 6 -12 months of age. However, in 1 year-old 3xTgAD mice, neuropathological changes, as defined by the presence of extracellular plaques and hyperphosphorylated tau, were very mild. Conclusions: The results suggest that: 1) 3xTgAD mice might develop a systemic autoimmune/inflammatory condition before severe learning/ memory deficits emerge. The lack of typical AD brain pathology and profound immune dysfunction suggest that changes in the immune system might be a factor influencing the development of MCI- and subsequent AD-like behavioral dysfunction in the 3xTgAD model 2) 3xTgAD mice display a MCI-like stage of disease development that may be useful in the further study of this phase of cognitive decline. P1-025
EVALUATING TNF-ALPHA AS A POSSIBLE LINK IN ALZHEIMER’S DISEASE AND RHEUMATOID ARTHRITIS
Evangelia Paouri1, Spiros Georgopoulos1, 1Biomedical Research Foundation, Academy of Athens, Athens, Greece. Contact e-mail: [email protected] Background: A number of studies have produced controversial data about the relation between Alzheimer’s disease (AD) and rheumatoid arthritis (RA). A number of inflammatory mediators including TNF- a are involved in both diseases. TNF- a is a potent pro-inflammatory cytokine involved in a number of pathologies including rheumatoid arthritis. Therapeutic approaches targeting TNF- a have been successful in the treatment of RA. TNF- a has been detected in AD human brains and is up-regulated in both the cerebrospinal fluid and serum of AD patients. However, the role of TNF- a in the course of AD is still unclear and its pathophysiological actions in AD have been reported to be controversial. In this study we evaluate TNF-a as a potent mediator in AD by inactivating TNF- a or over-expressing it in an AD mouse. Furthermore to investigate for a possible connection between AD and RA we used a TNF- a over-expressing transgenic mouse that develops RA. Methods: In the present study we inactivated the mouse TNF- a gene in a transgenic mouse model of AD, the 5xFAD transgenic mice by mating with TNF-a deficient mice. To overexpress TNF- a we used an established mouse of RA that over-express TNF- a in macrophages and develops arthritis, and mated it with the 5XFAD mice. Analysis was performed by using standard histopathological, immunohistochemical and biochemical methods. Results: Analysis of the phenotype in the 5XFAD mice that are TNF- a deficient or over-express TNF- a has revealed a significant effect on the amyloid phenotype. These data will be presented and analyzed in detail. Conclusions: Our data pinpoint the important role of TNF- a in AD, suggest TNF- a as a possible link between AD and RA, and support the use of anti-TNF a therapy in AD. P1-026
IRAK-M DELETION PROMOTES MICROGLIAMEDIATED BETA-AMYLOID PHAGOCYTOSIS IN PSAPP MICE
David Gate1, Gi-Bum Kim1, Joshua Breunig1, Eliezer Masliah2, Terrence Town3, 1Cedars-Sinai Medical Center, Los Angeles, California, United States; 2University of California San Diego, La Jolla, California, United States; 3Cedars-Sinai Medical Center/UCLA, Los Angeles, California, United States. Contact e-mail: [email protected] Background: Alzheimer disease (AD) is pathologically earmarked by deposition of amyloid-b (Ab) peptides as b-amyloid plaques, neuronal injury and low-level, chronic activation of brain innate immunity. It has been suggested that Ab engages toll-like receptors (TLRs), resulting in pro-
P1-023
NEUROINFLAMMATION IN HAPPSL TRANSGENIC MICE
Tina L€ offler1, Daniel Havas1, Stefanie Flunkert1, Nicole Taub1, Manfred Windisch2, Birgit Hutter-Paier1, 1QPS Austria, Grambach, Austria; 2QPS, Grambach, Austria. Contact e-mail: rigit. [email protected] Background: Deposition of neurotoxic amyloid beta peptides and the formation of neurofibrillary tangles represent the major pathological hallmarks of Alzheimer’s disease (AD). A rising body of evidence suggests neuroinflammation to be a third decisive component that actively contributes to disease progression and severity. Reactive astrocytes and microglia surrounding senile plaques play a pivotal role in the attempt to fight toxic depositions in AD brain, during which they can act as cytotoxic effector cells by releasing substances such as reactive oxygen species (ROS) and cytokines. These processes, oxidative stress and neuroinflammation, nourish the vicious circle of AD pathology and are thus seen as potential therapeutic targets. Methods: Cortical and hippocampal tissue of 6, 9 and 12 months old hAPP SL transgenic and non-transgenic mice was analyzed for soluble and insoluble Ab1-38, 1-40 and 1-42 with the MSDÒ 96-well MULTI-SPOTÒ 4G8 Ab Triplex Assay (Mesoscale Discovery). Brain tissue was histologically analyzed for astrocytosis and activated microglia by immunofluorescent staining with GFAP-IHC (Dako, Z0334) and CD11b-IHC (Serotec, MCA711) antibody, respectively. Furthermore, tissue of 4, 6, 9 and 12 months old animals was analyzed for oxidative stress by using the TBARS-assay. Results: Starting at an age of 6 months hAPP SL transgenic mice exhibit significantly enhanced Ab1-42 levels in the cortex and hippocampus followed by a significant increase of Ab1-38 and 1-40 at an age of 9 months. Simultaneously, neuroinflammation comprising microgliosis and astrocytosis increases in the cortex. Microgliosis also increases in the hippocampus and corpus callosum. Analysis of oxidative stress in 9 and 12 months old hAPP SL transgenic mice reveals a significant increase in the cortex and hippocampus compared to non-transgenic littermates and correlates with the higher incidence of glial inflammatory response. Conclusions: Next to a severe amyloid pathology, hAPP SL transgenic mice also develop neuroinflammation and oxidative stress at later time points, displaying a broad spectrum of AD pathology. These mice represent therefore a valid tool, not only to screen developmental compounds interfering with amyloid generation and plaque formation, but also anti-inflammatory and/ or immune-modulatory agents.
P1-024
IMMUNOLOGICAL DYSFUNCTION AND MCILIKE BEHAVIOR IN THE 3XTGAD MICE
Monica Marchese1, David Cowan1, Khalil Karimi1, Vanessa Ashthorpe1, Elizabeth Head2, Donglai Ma1, Hui Zhao1, Paulina Davis3, Minesh Kapadia1, Boris Sakic1, 1McMaster University, Hamilton, Ontario, Canada; 2Sanders-Brown Center on Aging, University of Kentucky, Lexington, Kentucky, United States; 3University of Kentucky, Lexington, Kentucky, United States. Contact e-mail: [email protected] Background: Although immune system activation has been observed in patients with Alzheimer’s disease (AD), it remains unclear whether inflammatory and/or autoimmune manifestations are epiphenomena, consequence, or cause of brain degeneration. We presently examine whether immunological changes accompany the progress of AD-like disease in well-established 3xTgAD murine model. Methods: We compared the immunological and behavioral profiles of triple transgenic mice (3xTgAD) and wild type (WT) controls, from 1.5 to 12 months of age. Mice underwent a broad battery of tests to assess longitudinal changes in behavioral performance.
P161
FACScan analysis and immunoenzymatic assays were employed to quantify inflammatory and autoimmune markers in serum and spleen. Results: Prior to the development of advanced AD pathology, 3xTgAD mice developed MCI-like manifestations, characterized by anxiety-related behaviors, changes in olfactory function and impaired flexibility in learning/memory tasks (2.5 months). Several manifestations of systemic inflammatory / autoimmune disease were observed at later age (12 months). They included splenomegaly, elevated serum levels of anti-nuclear and anti-dsDNA antibodies, as well as a reduced number of T splenocytes. Changes in the immune system occurred in parallel with generalized learning/memory and olfaction deficits from 6 -12 months of age. However, in 1 year-old 3xTgAD mice, neuropathological changes, as defined by the presence of extracellular plaques and hyperphosphorylated tau, were very mild. Conclusions: The results suggest that: 1) 3xTgAD mice might develop a systemic autoimmune/inflammatory condition before severe learning/ memory deficits emerge. The lack of typical AD brain pathology and profound immune dysfunction suggest that changes in the immune system might be a factor influencing the development of MCI- and subsequent AD-like behavioral dysfunction in the 3xTgAD model 2) 3xTgAD mice display a MCI-like stage of disease development that may be useful in the further study of this phase of cognitive decline. P1-025
EVALUATING TNF-ALPHA AS A POSSIBLE LINK IN ALZHEIMER’S DISEASE AND RHEUMATOID ARTHRITIS
Evangelia Paouri1, Spiros Georgopoulos1, 1Biomedical Research Foundation, Academy of Athens, Athens, Greece. Contact e-mail: [email protected] Background: A number of studies have produced controversial data about the relation between Alzheimer’s disease (AD) and rheumatoid arthritis (RA). A number of inflammatory mediators including TNF- a are involved in both diseases. TNF- a is a potent pro-inflammatory cytokine involved in a number of pathologies including rheumatoid arthritis. Therapeutic approaches targeting TNF- a have been successful in the treatment of RA. TNF- a has been detected in AD human brains and is up-regulated in both the cerebrospinal fluid and serum of AD patients. However, the role of TNF- a in the course of AD is still unclear and its pathophysiological actions in AD have been reported to be controversial. In this study we evaluate TNF-a as a potent mediator in AD by inactivating TNF- a or over-expressing it in an AD mouse. Furthermore to investigate for a possible connection between AD and RA we used a TNF- a over-expressing transgenic mouse that develops RA. Methods: In the present study we inactivated the mouse TNF- a gene in a transgenic mouse model of AD, the 5xFAD transgenic mice by mating with TNF-a deficient mice. To overexpress TNF- a we used an established mouse of RA that over-express TNF- a in macrophages and develops arthritis, and mated it with the 5XFAD mice. Analysis was performed by using standard histopathological, immunohistochemical and biochemical methods. Results: Analysis of the phenotype in the 5XFAD mice that are TNF- a deficient or over-express TNF- a has revealed a significant effect on the amyloid phenotype. These data will be presented and analyzed in detail. Conclusions: Our data pinpoint the important role of TNF- a in AD, suggest TNF- a as a possible link between AD and RA, and support the use of anti-TNF a therapy in AD. P1-026
IRAK-M DELETION PROMOTES MICROGLIAMEDIATED BETA-AMYLOID PHAGOCYTOSIS IN PSAPP MICE
David Gate1, Gi-Bum Kim1, Joshua Breunig1, Eliezer Masliah2, Terrence Town3, 1Cedars-Sinai Medical Center, Los Angeles, California, United States; 2University of California San Diego, La Jolla, California, United States; 3Cedars-Sinai Medical Center/UCLA, Los Angeles, California, United States. Contact e-mail: [email protected] Background: Alzheimer disease (AD) is pathologically earmarked by deposition of amyloid-b (Ab) peptides as b-amyloid plaques, neuronal injury and low-level, chronic activation of brain innate immunity. It has been suggested that Ab engages toll-like receptors (TLRs), resulting in pro-