Sunday, July 14, 2013: Poster Presentations: P1P160

disorders like Alzheimer’s disease because the disease and treatments often

directly modulate attention and alertness. Methods: Subjects included 8

male long evans rats who were acclimated to the restraint and noise in

a mock scanner. The MRI session consisted of a 7 min structural scan

followed by a 10 min baseline fMRI scan. At that point, either IGF2

(30 mg/kg recombinant IGF2) or Vehicle (0.01% BSA in sterile PBS) was

administered intravenously and pharmacological MRI response was

acquired for the following 25 minutes. Results: Compared to vehicle,

IGF2 modulated activity in multiple regions impacted by Alzheimer’s dis-

ease, including frontal cortex, basal forebrain, and hippocampal regions.

Conclusions: Our preliminary findings at a single dose further support

the potential utility of IGF2 as a therapeutic for Alzheimer’s disease.

P1-020 CALORIE RESTRICTION, BUT NOT KETOGENIC

DIET, IMPROVES COGNITION IN MOUSE

MODELS OFALZHEIMER’S PATHOLOGY

Milene Brownlow1, Leif Benner2, Aurelie Joly-Amado1, Sana Azam1,

Dominic D’Agostino1, Marcia N. Gordon1, David Morgan1, 1University of

South Florida, Tampa, Florida, United States; 2University of Tampa,

Tampa, Florida, United States. Contact e-mail: mbrownlo@health.usf.edu

Background: Calorie restriction (CR) and Ketogenic Diets (KDs) have

been recently studied as possible therapeutic approaches for neurodegener-

ative diseases. CR has been previously shown to efficiently improve cogni-

tion and decreased pathology in amyloid depositing mouse models of

Alzheimer’s disease (AD). KDs, widely investigated in the treatment and

prevention of seizures, have been suggested to bypass metabolic deficits

present in AD brains by providing ketone bodies as an alternative fuel,

once impairments in glucose/insulin pathways take place. Methods: We

investigated the effects of a KD on both amyloid (APP+PS1) and tau depos-

iting models (Tg4510) of AD and the effects of CR in the tau depositing

mouse model. Body weight and food intake were monitored throughout

the studies. Blood was collected for ketone and glucose assessments. After

being on their respective diets for 3months,micewere submitted to a battery

of behavioral tests.Results: In the KD study, we found increased locomotor

activity in the open field was also observed in both transgenic models

studied, compared to age-matched nontransgenic controls. KD-fed mice

performed significantly better on the rotarod test compared to mice on the

control diet irrespective of genotype, showing an improved motor perfor-

mance. No changes in cognition or histopathological markers were observed

in mice fed KD, compared to control diet. However, Tg4510mice submitted

to a 35% body weight reduction over 3 months (CR) performed significantly

better in the novel object recognition test, suggesting improved memory,

compared to transgenic mice fed ad libitum. Histopathological assessments

of tau and microglial markers are currently ongoing. Conclusions: This

initial data suggests that: a) KD may play an important role in motor perfor-

mance in mice possibly through peripheral mechanisms; b) CR significantly

rescued memory deficits in tau depositing mice.

P1-021 DEVELOPMENT OFA SPORADIC ANIMAL

MODEL OFALZHEIMER’S DISEASE

Frank Sharp, Xinhua Zhan,University of California at Davis, Sacramento,

California, United States. Contact e-mail: frank.sharp@ucdmc.ucdavis.edu

Background: Ischemia, white matter, and Alzheimer’s Disease (AD)

pathologies often co-exist in aging human brain. It is not known if one

can cause or contribute to the other.Methods: To search for a link between

them, 3 month old adult rats were administered Lipopolysacchardide (LPS,

3mg/kg intraperitoneally) and 24h later subjected to 20 minutes of transient

focal middle cerebral artery ischemia using the suture method. 30 minutes

later animals were subjected to 30 minutes of hypoxia (8% oxygen). This

LPS/Ischemia/Hypoxia (LPS/IS/H) model has previously been studied in

neonates but not in adults. Brains were examined for myelin/axon damage,

cytokines and abeta/amyloid deposition, and the findings compared to the

5XFAD mouse AD model. Results: Using an antibody that detected intact

myelin basic protein (MBP) and degraded myelin basic protein (dMBP),

MBP was markedly depleted in the ischemic hemisphere. In addition,

dMBP was detected bilaterally in the ischemic and non-ischemic hemi-

spheres, with dMBP increasing at 12 weeks compared to 4 weeks. The

increasing dMBP levels correlated with increasing levels of the cytokines

IL1beta and Granzyme B in cortex. MBP immunocytochemistry showed

that MBP localized to axons in normal control cortex, but at 1 week post

LPS/IS/H it localized to blood vessel endothelial cells in cortex and hippo-

campus bilaterally. From 2 to 8 weeks post LPS/IS/H there were foci of

tangled dysmorphic myelinated axons that were found throughout cortex

and hippocampus bilaterally. Ab and amyloid deposition occurred in myelin

aggregates in w1/3 of subjects at 8 and 12 weeks in cortex in the ischemic

hemisphere (Figure 1). Thioflavin S, silver stained and FSB stained amyloid

plaques were detected 8-12 weeks following LPS/IS/H. Ab deposition and

amyloid plaques in the 5XFAD mouse were associated with myelin aggre-

gates throughout cortex. Conclusions: Lipopolysaccharide/Ischemia/

Hypoxia produces elements of white matter and AD pathologies in adult

brain. Myelin aggregates form in the cortex following LPS/IS/H and in

the 5XFAD mouse AD model, and are proposed to form the nidus for abeta

and amyloid deposition that results in amyloid plaques in both.

P1-022 MODULATION OF PERIPHERAL IMMUNE CELL

TRAFFIC TO THE CNS BY XPRO1595 IN A MOUSE

MODEL OFALZHEIMER’S DISEASE

Kathryn MacPherson, Malu Tansey, Emory University School of

Medicine, Atlanta, Georgia, United States. Contact e-mail:

kpmacpherson@gmail.com

Background: Clinically, systemic infection is associated with development

of dementia in elderly patients and increased cognitive decline in AD

patients. Epidemiological studies have found older people who use anti-in-

flammatory drugs regularly have lower incidence of AD. These observations

prompt the hypotheses that (1) the peripheral immune system plays an active

role in AD and (2) targeting specific elements of peripheral inflammation

could be useful in treating disease. In support of this, chronic peripheral

administration of lipopolysaccharide (LPS) increases central inflammation

(including Tumor Necrosis Factor, TNF), is associated with enhanced cog-

nitive decline and primedmicroglia. Soluble TNF is known to increase BBB

leakiness possibly leading to increased peripheral immune traffic to the CNS

in conditions of chronic systemic inflammation. Targeted inhibition of TNF

in the CNS can ameliorate pathology induced by systemic inflammation.

However, the specific role of soluble TNF in systemic circulation and its

contribution to peripheral immune cell infiltration in AD has not been ex-

plored. Methods: We characterized the inflammatory profile of 5xFAD

mice, a transgenic mouse with early onset of amyloid plaques, memory

impairment, and neurodegeneration. Effects of peripheral LPS and the

TNF inhibitor XPro1595 administration on AD progression were assessed.

Memory impairment was measured via the novel object recognition task.

Quantification of gene and protein expression level was used to track AD

progression and neuroinflammation. Flow cytometry was used to count im-

mune cells present in the CNS during disease progression. Results: In the

CNS we detected increased levels of CD45 at 4 and 6 months of age and in-

creased CCL2 and TNF at 6 months of age in 5xFAD mice versus non-Tg

mice. At 6 months of age increased levels of neutrophils and monocytes

Sunday, July 14, 2013: Poster Presentations: P1 P161

were found in 5xFAD versus non-Tg mice. Conclusions: Preliminary re-

sults 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 (cen-

tral 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.

hutter-paier@qps.com

Background: Deposition of neurotoxic amyloid beta peptides and the for-

mation of neurofibrillary tangles represent the major pathological hallmarks

of Alzheimer’s disease (AD). A rising body of evidence suggests neuroin-

flammation to be a third decisive component that actively contributes to

disease progression and severity. Reactive astrocytes and microglia sur-

rounding 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 cy-

tokines. 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 12months old

hAPP SL transgenic and non-transgenic mice was analyzed for soluble and

insoluble Ab1-38, 1-40 and 1-42 with the MSD� 96-well MULTI-SPOT�4G8 Ab Triplex Assay (Mesoscale Discovery). Brain tissue was histologi-

cally analyzed for astrocytosis and activated microglia by immunofluores-

cent 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 hippo-

campus 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 hippo-

campus 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 corre-

lates with the higher incidence of glial inflammatory response. Conclu-

sions: Next to a severe amyloid pathology, hAPP SL transgenic mice also

develop neuroinflammation and oxidative stress at later time points, display-

ing 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 MCI-

LIKE 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: marches@mcmaster.ca

Background: Although immune system activation has been observed in

patients with Alzheimer’s disease (AD), it remains unclear whether inflam-

matory and/or autoimmunemanifestations are epiphenomena, consequence,

or cause of brain degeneration. We presently examine whether immunolog-

ical 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.

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 / auto-

immune disease were observed at later age (12 months). They included

splenomegaly, elevated serum levels of anti-nuclear and anti-dsDNA anti-

bodies, 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 pro-

found 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 3xTgADmodel 2) 3xTgADmice dis-

play a MCI-like stage of disease development that may be useful in the fur-

ther 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:

sgeorgopoulos@bioacademy.gr

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 ADmouse. Furthermore to investigate for a possible

connection between AD and RA we used a TNF- a over-expressing trans-

genic mouse that develops RA. Methods: In the present study we inacti-

vated the mouse TNF- a gene in a transgenic mouse model of AD, the

5xFAD transgenic mice by mating with TNF-a deficient mice. To over-

express 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 pin-

point 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 MICROGLIA-

MEDIATED 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: david.gate@cshs.org

Background: Alzheimer disease (AD) is pathologically earmarked by de-

position of amyloid-b (Ab) peptides as b-amyloid plaques, neuronal injury

and low-level, chronic activation of brain innate immunity. It has been sug-

gested that Ab engages toll-like receptors (TLRs), resulting in pro-