- Email: [email protected]
IRAK-M deletion promotes microglia-mediated beta-amyloid phagocytosis in PSAPP mice
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-
P162
Sunday, July 14, 2013: Poster Presentations: P1
inflammatory activation of microglia. TLR mediated microglial activation can be beneficial by stimulating phagocytosis but can also be deleterious by endorsing Ab-stimulated release of neurotoxic molecules. TLRs transduce their signals through MyD88 and the serine/threonine IL-1 receptorassociated kinase (IRAK). The IRAK family of kinases generally upregulate TLR signaling, with the notable exception of the inhibitory kinase IRAK-M. IRAK-M expression is specific to cells of monocytic lineage, including microglia, and plays a critical role in the maintenance of innate immune homeostasis by dampening inflammation. Methods: We crossed mice deficient in IRAK-M (IRAK-M -/-) with mice over expressing mutant human amyloid precursor protein, the PSAPP mouse model of cerebral amyloidosis. Innate immune cell activation and cerebral A b burden were subsequently evaluated in age- and sex-matched littermates from PSAPPIRAK-M + and PSAPP- IRAK-M -/- mice at 15 and 25 months. Results: Immunohistochemistry for the reactive microglial marker Iba1 disclosed statistically significant increased expression in PSAPP- IRAK-M -/- vs. PSAPP- IRAK-M + mice in the hippocampus, cerebral cortex and entorhinal cortex (up 38-42%), brain regions associated with AD-type pathology in humans. Further, brain parenchymal Ab abundance was correspondingly attenuated in PSAPP- IRAK-M -/- mice as measured by ThioS staining and ELISA. Finally, in vivo quantification of Ab within Lamp2 + phagolysosomes revealed a microglial phagocytic mechanism of Ab clearance in IRAK-M knockouts. Conclusions: These data suggest that IRAK-M negatively regulates brain innate immunity in a mouse model of cerebral amyloidosis. Collectively then, disinhibition of IRAK-M represents a means to target migroglial remodeling and clearance of amyloid pathology.
P1-027
INTRACRANIAL ADMINISTRATION OF GAMMAGARD IVIG LOWERS AMYLOID AND MODULATES NEUROINFLAMMATORY PROFILES ALONG A DIFFERENT TIME-COURSE THAN ANTI-BETA-AMYLOID IGG: IMPLICATIONS FOR MECHANISM OF ACTION
Donna Wilcock, Tiffany Sudduth, Abigail Greenstein, University of Kentucky, Lexington, Kentucky, United States. Contact e-mail: donna. [email protected] Background: Gammargard IVIg is a therapeutic approach to treat Alzheimer’s disease currently in phase 3 clinical trials. Despite the reported efficacy of the approach, the mechanism of action is poorly understood. We have previously shown that intracranial injection of anti-Abantibodies into the frontal cortex and hippocampus reveals important information regarding the time-course of events once the agent is in the brain. In the current study we compared Gammagard IVIg, mouse pooled IgG and the anti-A b antibody 6E10. Methods: We injected Gammagard IVIg, mouse pooled IgG and the anti-A b antibody 6E10 intracranially into the frontal cortex and hippocampus of 9 month old APP/PS1 mice. We established a time-course of events ranging from 24 hours to 21 days post-injection. Results: Gammagard IVIg and pooled mouse IgG both significantly reduced amyloid deposition to the same degree as the 6E10 anti-Abantibody, however, the clearance was much slower to occur, happening between the 3 day and 7 day time-points. In contrast, as we have previously shown, amyloid reductions were apparent with the 6E10 anti-Abgroup at the 1-day time-point. Also, neuroinflammatory profiles were significantly altered by the antibody treatments. APP/PS1 transgenic mice at nine months of age typically exhibit an M2a inflammatory phenotype. Anti-AbIgG stimulated a brief M2b state, peaking at 2 days and resolving completely by 7 days. In contrast, Gammagard IVIg and pooled mouse IgG both stimulated an M2b response that peaked at 7 days and was sustained through the 14 day time-point, only resolving by 21 days. Conclusions: Because the neuroinflammatory switch occurs prior to the detectable reductions in amyloid deposition, we hypothesize that the Gammagard IVIg and pooled mouse IgG act as immune modulators and this immune modulation is responsible for the reductions in amyloid pathology. Future studies will focus on this mechanism of action to determine whether immune modulation avoids the cerebrovascular adverse events that have plagued the anti-Ab immunotherapy approaches.
P1-028
PREVENTIVE EFFECTS OF CARDIOTONIC PILLS ON WHITE MATTER DAMAGE AND NEUROINFLAMMATION INDUCED BY CHRONIC CEREBRAL HYPOPERFUSION IN A RAT MODEL
WonKyung Jeon1, Jihye Bang2, Ki-Mo Lee3, 1Korea Institute of Oriental Medicine, Daejeon, South Korea; 2Korea Institute of Oriental Medicine, Daejeon, South Korea; 3Herbal Medicine Research Division, Daejeon, South Korea. Contact e-mail: [email protected] Background: Cardiotonic pills (CP) is a commercial herbal medicine including Salvia miltiorrhiza, Panax notoginseng and Dryobalanops Aromatica Gaertner and is widely used to treat cardiovascular diseases. We investigated the effects of CP in rats with chronic cerebral hypoperfusion. Methods: Chronic cerebral hypoperfusion was induced in male Wister rats by permanent bilateral common carotid artery occlusion (BCCAo). Daily administration of CP (200 mg/kg) was started on day 20 after BCCAo and continued for 42 days. Immunohistochemical analysis was performed in basal forebrain and white matter of BCCAo-induced rats by using myelin basic protein (MBP), Iba-1, OX-6 and choline acetyltransferase (ChAT) antibody. The expression level of mitogen-activated protein kinases (MAPKs), interleukin-1 b (IL-1 b), interleukin-6 (IL-6), cyclooxygenase2 (Cox-2), ChAT and MBP in the BCCAo brains was analyzed by Western blot. Results: In the white matter regions, treatment with CP improved the MBP levels decreased by chronic cerebral hypoperfusion and the levels of microglial activation mitigated (vehicle-treated BCCAo; 11.5 62.8 and CP-treated BCCAo; 9.062.3, P<0.05 in corpus callosum, vehicle; 19.3 65.4 and CP; 9.062.3, P<0.05 in fimbria, vehicle; 43.9 620.7 and CP; 24.5611.4, P<0.05 in optic tract, vehicle; 30.7 64.7 and CP; 22.765.1, P<0.01 in hippocampus, respectively). In the basal forebrain, the number of cholinergic neurons were increased by CP treatment (vehicle; 17.0 65.0 and CP; 21.066.0, P ¼0.063). In addition, CP significantly suppressed the phosphorylation of MAPK and ameliorated the altered expression of IL-1 b, IL-6, ChAT and MBP in the BCCAo hippocampus and white matter regions. Conclusions: These results indicate that CP may have therapeutic potential for the prevention of vascular dementia via inhibition of inflammatory processes. P1-029
THE EFFECT OF MELATONIN ON LIFESPAN IN THE MODEL ORGANISM CAENORHABDITIS ELEGANS
Omer Karadas1, Hakan Gul2, Necati Ozpinar1, Serdar Firtina1, 1Erzincan Military Hospital, Erzincan, Turkey; 2Kartal Education and Treatment Hospital, Istanbul, Turkey. Contact e-mail: [email protected] Background: The objective of this experimental study was to evaluate the physiological and lifespan effects of different doses of melatonin on animal model; caenorhabditis elegans. Methods: Different doses of melatonin that has high antioxidant activity has been tested on c.elegans. In this study; 25, 50, 100, 250, 500 and 1000 mg/ml concentrations of melatonin was prepared and tested. Concentrations with the highest effects were detected. Results: 25, 50, 100 mg/ml of melatonin effects lifespan and fertiliy of c.elegans. This data resulting from our study can be a step for the future studies about the biological activity. Conclusions: Some molecules have antioxidant activity. Melatonin as an antioxidant is known as the terminal antioxidant but it has not prooxidant activity. It is an important protector against the cellular toxicity. Melatonin’s positive effect on aging and healthy lifestyle is significant. Antioxidants used on caenorhabditis elegans (c. Elegans) have been shown to prolong the lifetime and termotolerans. P1-030
AMELIORATION OF ALZHEIMER’S DISEASE BY THE NEUROPROTECTIVE EFFECT OF SULFORAPHANE IN AN ANIMAL MODEL
Hye Yun Kim1, Hyunjin Vincent Kim1, Hanna Ehrlich2, Seon Young Choi1, Dong Jin Kim1, YoungSoo Kim1, 1KIST, Seoul, South Korea; 2Tufts University, Boston, Massachusetts, United States. Contact e-mail: [email protected]
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-
P162
Sunday, July 14, 2013: Poster Presentations: P1
inflammatory activation of microglia. TLR mediated microglial activation can be beneficial by stimulating phagocytosis but can also be deleterious by endorsing Ab-stimulated release of neurotoxic molecules. TLRs transduce their signals through MyD88 and the serine/threonine IL-1 receptorassociated kinase (IRAK). The IRAK family of kinases generally upregulate TLR signaling, with the notable exception of the inhibitory kinase IRAK-M. IRAK-M expression is specific to cells of monocytic lineage, including microglia, and plays a critical role in the maintenance of innate immune homeostasis by dampening inflammation. Methods: We crossed mice deficient in IRAK-M (IRAK-M -/-) with mice over expressing mutant human amyloid precursor protein, the PSAPP mouse model of cerebral amyloidosis. Innate immune cell activation and cerebral A b burden were subsequently evaluated in age- and sex-matched littermates from PSAPPIRAK-M + and PSAPP- IRAK-M -/- mice at 15 and 25 months. Results: Immunohistochemistry for the reactive microglial marker Iba1 disclosed statistically significant increased expression in PSAPP- IRAK-M -/- vs. PSAPP- IRAK-M + mice in the hippocampus, cerebral cortex and entorhinal cortex (up 38-42%), brain regions associated with AD-type pathology in humans. Further, brain parenchymal Ab abundance was correspondingly attenuated in PSAPP- IRAK-M -/- mice as measured by ThioS staining and ELISA. Finally, in vivo quantification of Ab within Lamp2 + phagolysosomes revealed a microglial phagocytic mechanism of Ab clearance in IRAK-M knockouts. Conclusions: These data suggest that IRAK-M negatively regulates brain innate immunity in a mouse model of cerebral amyloidosis. Collectively then, disinhibition of IRAK-M represents a means to target migroglial remodeling and clearance of amyloid pathology.
P1-027
INTRACRANIAL ADMINISTRATION OF GAMMAGARD IVIG LOWERS AMYLOID AND MODULATES NEUROINFLAMMATORY PROFILES ALONG A DIFFERENT TIME-COURSE THAN ANTI-BETA-AMYLOID IGG: IMPLICATIONS FOR MECHANISM OF ACTION
Donna Wilcock, Tiffany Sudduth, Abigail Greenstein, University of Kentucky, Lexington, Kentucky, United States. Contact e-mail: donna. [email protected] Background: Gammargard IVIg is a therapeutic approach to treat Alzheimer’s disease currently in phase 3 clinical trials. Despite the reported efficacy of the approach, the mechanism of action is poorly understood. We have previously shown that intracranial injection of anti-Abantibodies into the frontal cortex and hippocampus reveals important information regarding the time-course of events once the agent is in the brain. In the current study we compared Gammagard IVIg, mouse pooled IgG and the anti-A b antibody 6E10. Methods: We injected Gammagard IVIg, mouse pooled IgG and the anti-A b antibody 6E10 intracranially into the frontal cortex and hippocampus of 9 month old APP/PS1 mice. We established a time-course of events ranging from 24 hours to 21 days post-injection. Results: Gammagard IVIg and pooled mouse IgG both significantly reduced amyloid deposition to the same degree as the 6E10 anti-Abantibody, however, the clearance was much slower to occur, happening between the 3 day and 7 day time-points. In contrast, as we have previously shown, amyloid reductions were apparent with the 6E10 anti-Abgroup at the 1-day time-point. Also, neuroinflammatory profiles were significantly altered by the antibody treatments. APP/PS1 transgenic mice at nine months of age typically exhibit an M2a inflammatory phenotype. Anti-AbIgG stimulated a brief M2b state, peaking at 2 days and resolving completely by 7 days. In contrast, Gammagard IVIg and pooled mouse IgG both stimulated an M2b response that peaked at 7 days and was sustained through the 14 day time-point, only resolving by 21 days. Conclusions: Because the neuroinflammatory switch occurs prior to the detectable reductions in amyloid deposition, we hypothesize that the Gammagard IVIg and pooled mouse IgG act as immune modulators and this immune modulation is responsible for the reductions in amyloid pathology. Future studies will focus on this mechanism of action to determine whether immune modulation avoids the cerebrovascular adverse events that have plagued the anti-Ab immunotherapy approaches.
P1-028
PREVENTIVE EFFECTS OF CARDIOTONIC PILLS ON WHITE MATTER DAMAGE AND NEUROINFLAMMATION INDUCED BY CHRONIC CEREBRAL HYPOPERFUSION IN A RAT MODEL
WonKyung Jeon1, Jihye Bang2, Ki-Mo Lee3, 1Korea Institute of Oriental Medicine, Daejeon, South Korea; 2Korea Institute of Oriental Medicine, Daejeon, South Korea; 3Herbal Medicine Research Division, Daejeon, South Korea. Contact e-mail: [email protected] Background: Cardiotonic pills (CP) is a commercial herbal medicine including Salvia miltiorrhiza, Panax notoginseng and Dryobalanops Aromatica Gaertner and is widely used to treat cardiovascular diseases. We investigated the effects of CP in rats with chronic cerebral hypoperfusion. Methods: Chronic cerebral hypoperfusion was induced in male Wister rats by permanent bilateral common carotid artery occlusion (BCCAo). Daily administration of CP (200 mg/kg) was started on day 20 after BCCAo and continued for 42 days. Immunohistochemical analysis was performed in basal forebrain and white matter of BCCAo-induced rats by using myelin basic protein (MBP), Iba-1, OX-6 and choline acetyltransferase (ChAT) antibody. The expression level of mitogen-activated protein kinases (MAPKs), interleukin-1 b (IL-1 b), interleukin-6 (IL-6), cyclooxygenase2 (Cox-2), ChAT and MBP in the BCCAo brains was analyzed by Western blot. Results: In the white matter regions, treatment with CP improved the MBP levels decreased by chronic cerebral hypoperfusion and the levels of microglial activation mitigated (vehicle-treated BCCAo; 11.5 62.8 and CP-treated BCCAo; 9.062.3, P<0.05 in corpus callosum, vehicle; 19.3 65.4 and CP; 9.062.3, P<0.05 in fimbria, vehicle; 43.9 620.7 and CP; 24.5611.4, P<0.05 in optic tract, vehicle; 30.7 64.7 and CP; 22.765.1, P<0.01 in hippocampus, respectively). In the basal forebrain, the number of cholinergic neurons were increased by CP treatment (vehicle; 17.0 65.0 and CP; 21.066.0, P ¼0.063). In addition, CP significantly suppressed the phosphorylation of MAPK and ameliorated the altered expression of IL-1 b, IL-6, ChAT and MBP in the BCCAo hippocampus and white matter regions. Conclusions: These results indicate that CP may have therapeutic potential for the prevention of vascular dementia via inhibition of inflammatory processes. P1-029
THE EFFECT OF MELATONIN ON LIFESPAN IN THE MODEL ORGANISM CAENORHABDITIS ELEGANS
Omer Karadas1, Hakan Gul2, Necati Ozpinar1, Serdar Firtina1, 1Erzincan Military Hospital, Erzincan, Turkey; 2Kartal Education and Treatment Hospital, Istanbul, Turkey. Contact e-mail: [email protected] Background: The objective of this experimental study was to evaluate the physiological and lifespan effects of different doses of melatonin on animal model; caenorhabditis elegans. Methods: Different doses of melatonin that has high antioxidant activity has been tested on c.elegans. In this study; 25, 50, 100, 250, 500 and 1000 mg/ml concentrations of melatonin was prepared and tested. Concentrations with the highest effects were detected. Results: 25, 50, 100 mg/ml of melatonin effects lifespan and fertiliy of c.elegans. This data resulting from our study can be a step for the future studies about the biological activity. Conclusions: Some molecules have antioxidant activity. Melatonin as an antioxidant is known as the terminal antioxidant but it has not prooxidant activity. It is an important protector against the cellular toxicity. Melatonin’s positive effect on aging and healthy lifestyle is significant. Antioxidants used on caenorhabditis elegans (c. Elegans) have been shown to prolong the lifetime and termotolerans. P1-030
AMELIORATION OF ALZHEIMER’S DISEASE BY THE NEUROPROTECTIVE EFFECT OF SULFORAPHANE IN AN ANIMAL MODEL
Hye Yun Kim1, Hyunjin Vincent Kim1, Hanna Ehrlich2, Seon Young Choi1, Dong Jin Kim1, YoungSoo Kim1, 1KIST, Seoul, South Korea; 2Tufts University, Boston, Massachusetts, United States. Contact e-mail: [email protected]