Alzheimer’s disease (AD) is a neurodegenerative disorder primarily affecting regions of the brain responsible for higher cognitive
disease patients. Here, we further investigated their physiological role: in epitope mapping studies, NAbs–A? recognized the mid-/C-
peptide toxicity, but NAbs–A? did not readily clear senile plaques although early fleecy-like plaques were reduced. Administration of
mice. These findings suggest a novel physiological mechanism involving NAbs–A? to dispose of proteins or peptides that are prone to
Alzheimer’s disease (AD) is a neurodegenerative disorder pri-
marily affecting regions of the brain responsible for higher cog-
and neurofibrillary tangles are the histopathological hallmarks
of AD. ?-Amyloid (A?) peptides become deposited in those
plaques, and hence their clearance has been discussed as a major
Emerging experimental evidence detected the early acidic en-
et al., 2006; Rajendran et al., 2006), and dimers, trimers, and
multimeric aggregates have been shown recently in vitro and in
al., 2005; Klyubin et al., 2005; Lesne ´ et al., 2006; Townsend et al.,
further suggested that small A? oligomers may form intracellularly
before being released into the extracellular medium, in which they
venting or reversing the formation of aggregated amyloid would
Several therapeutic approaches are currently under consider-
ation, including active/passive immunization against A? as pio-
neered by Schenk et al. (1999). In transgenic amyloid precursor
protein (APP)-expressing mice, immunization against A? pep-
tides has been shown to be effective on both molecular and be-
havioral levels. Active immunization in transgenic mice reduced
fered with tau phosphorylation (Schenk et al., 1999; Morgan et al.,
2000). Moreover, passive immunization was also effective with an-
tibodies that recognized the N-terminal and the mid-terminal do-
mains of A? peptides (DeMattos et al., 2001). Based on these data,
patients treated with antibodies directed against the N terminus of
A?, a considerable decrease in plaque load has been reported, but
clearance of already formed plaques might not be sufficient to im-
Recently, we and others identified naturally occurring auto-
AD (Du et al., 2001; Weksler et al., 2002). Naturally occurring
autoantibodies make up to two-thirds of the human antibody
pool and are known to have many functions; however, the un-
derlying mechanisms are far from being completely understood
(Shoenfeld et al., 2007). NAbs–A? have been characterized in
(UKGM) (PN: 155/06-07MR) and in part by an unrestricted grant of the Deutsche Forschungsgemeinschaft Grant
DO784/2-1. We also received an unrestricted grant from Prof. Dr. Reinfried Pohl, Marburg, Germany. We thank
TheJournalofNeuroscience,April13,2011 • 31(15):5847–5854 • 5847
different experimental settings to inhibit the propensity of A? to
fibrillize, thereby blocking its toxicity and to affect the clearance
Relkin et al., 2009). However, how NAbs–A? interact with A?
and promote their clearance remains to be elucidated. Here, we
show for the first time that NAbs–A? interfered with, and
preferentially bound to early oligomerization products of A?
was reduced after passive immunization with NAbs–A? and
the subsequent clearance of A? led to a rapid improvement of
Based on the concept of NAbs–A?, commercially available
for the treatment of patients with AD (Dodel et al., 2010), which
trial in the United States (Relkin et al., 2009).
Isolation of NAbs–A?. We used purified human intravenous IgG (Oc-
tagam 5%) for the isolation of NAbs–A?, which was kindly provided by
plasma (10,000–20,000 of donations). The product is prepared by using
tion and chromatography and contains ?50 mg of protein per milliliter
(5%). Ninety-six percent of protein represents human normal IgG
(IgA ? 0.2 mg; IgM ? 0.1 mg). It contains no more than 3% aggregates
and fragments, respectively, and no more than 90% monomers and
dimers with an Fc portion maintained intact. IgG subclasses are fully
represented (IgG 1, 65%; IgG 2, 30%; IgG 3, 3%; IgG 4, 2%). For the
following isolation steps, this preparation was mixed with an equal vol-
ume of PBS and loaded directly onto an affinity column.
Because the A?1-40sequence contains internal lysine residues that
might lead to side reactions in immobilization procedures using amino
groups, a specific affinity column was prepared using a cysteine residue
attached to the A? N terminus to ensure homogeneous orientation of
peptide molecules on the column support by immobilization through
cysteinyl-S-thioether linkage. The azlactone-activated support contains
an iodoacetyl group (Ultralink; Perbio) at the end of a hexadecyl-spacer
group, which reacts with the cysteinyl-sulfhydryl group to yield a stable
thioether linkage to reduce steric hindrance and provide maximum
binding capacity of the antibodies. For covalent attachment of the Cys–
A?1-40, 3.7 mg of peptide was dissolved in 50 mM Tris and 5 mM
EDTA-Na coupling buffer, pH 8.5, to a final concentration of 0.37 mg/
ml. The solution was added to 1 ml of drained Ultralink-Iodoacetyl gel,
and the coupling reaction was performed for 1 h at room temperature
under gentle mixing, followed by 30 min reaction time without mixing.
An aliquot of 0.5 ml of the Cys–A?1-40coupled support was packed into
a column (2.5 ml; MoBiTec) allowing the solution to drain. The column
HCl in coupling buffer. Subsequently, the column was washed with 5 ml
of 1 M NaCl, 5 ml of 0.l M Na-phosphate, and 0.15 M NaCl, pH 7.2, and
stored at 4°C. The gel support (0.5 ml) was transferred into a 15 ml
Falcon vial using 5 ml of PBS and mixed with 5 ml of IVIg. After gentle
shaking overnight at 4°C, this suspension was transferred to the column
using the effluent to completely rinse the matrix back into the column.
The column was washed eight times with 10 ml of PBS, followed by two
wash cycles with 10 ml of ultrapure water.
The bound antibodies were eluted from the column with 10 ? 0.5 ml
of 0.1 M glycine buffer, pH 2.8. Each fraction was collected in a microre-
action tube containing 35 ?l of 1 M Tris-HCl, pH 9. The flow-through
(FT) of this isolation procedure, which contains IgG depleted of NAbs–
A?, was also collected and was used as a control in the respective exper-
To maintain the integrity of the antibodies, a neutral pH was adjusted
or glycine buffer. To regenerate the column for additional use, the col-
followed by two wash cycles with 10 ml of PBS containing 1 M NaCl and
finally two wash cycles with 10 ml of PBS.
Antibody concentrations in the elution fractions were determined by
the Micro BCA Protein Assay kit method (Pierce; Perbio). The stock
solution of 2 mg/ml bovine albumin supplied within the Micro BCATM
kit was used to prepare fresh standard dilutions within the range 40–0.5
?g/ml. The antibodies were eluted between fractions 1 and 6, with the
of 10 elutions, fresh albumin standard dilutions were prepared. Results
were read at 562 nm with an ELISA reader.
Cell culture and toxicity assay. SH-SY5Y human neuroblastoma cells
HEPES, 4 mM glutamine, penicillin (200 U/ml), and streptomycin (200
?g/ml) at 37°C in a 5% CO2atmosphere. Cells were plated and kept
After removal of medium, cells were washed with PBS, and toxic A?
oligomers (2 ?M final concentration) (Kayed et al., 2003) were added in
100 ?l of fresh medium with either 7.5 or 15 ?M NAbs–A? or without
Preparation of A?. The lyophilized synthetic A?1-40(PSL GmbH) was
resuspended in 100% 1,1,1,3,3,3 hexafluoro-2-propanol (HFIP) at 1
mg/ml and incubated for 1 h at room temperature. After evaporation of
HFIP, the peptide was resuspended according to the species of interest.
For preparation of the monomer, the synthetic A?1-40peptide was sus-
pended in ultrapure or double-distilled water at 1 mg/ml just before the
experiment. A? dimers were prepared using A?1-40peptide with
N-terminal cysteine (Cys–A?1-40) in 1? PBS at 1 mg/ml concentration
and centrifuged for 30 min at 12,000 rpm. The protein content of the
supernatant was quantified using Nanodrop and sonicated for 15 min at
(900 rpm). The trimer was generated by a double mutation of A?1-40
blastoma SH-SY5Y cells. SH-SY5Y cells (30,000 cells per well) were incubated alone or in the
determinations from a representative experiment repeated at least three times with similar
5848 • J.Neurosci.,April13,2011 • 31(15):5847–5854Dodeletal.•NaturallyOccurringAntibodiesandAlzheimer’sDisease
after the HFIP treatment, which instantly formed the SDS-stable trimer.
Low-binding minicentrifuge tubes were used (Eppendorf) throughout
Characterization of A? epitopes recognized by NAbs–A?. Ninety-six-
well plates were coated with A?1-40and other peptides as outlined in the
NaH2PO4, 98 mM Na2HPO4, pH 7.4, and 0.05% sodium azide)], and
incubated at 37°C for 2 h. The plates were then incubated with blocking
buffer (Pierce) at 37°C for 2 h.
1 h at 37°C (Calbiochem). The reaction with a chromogenic substrate,
3,3?,5,5?-tetramethylbenzidine (Calbiochem), was terminated with 2N
H2SO4, and plates were read at 450 nm using a plate reader (Thermo
Size exclusion chromatography. A Superdex 75 10/30 HR column at-
iments and calibrated using unbranched dextran standards of the
following molecular masses: 123,600; 66,700; 43,800; 21,400; 9890; and
4440 Da (Pharmacosmos). ?-Amyloid (1 mg/ml) was injected onto the
at a flow rate of 1 ml/min. One milliliter fractions were lyophilized,
resuspended in 2? SDS-tricine sample buffer (20 ?l), boiled for 10 min,
and then subjected to electrophoresis on 10–20% tricine gels. Proteins
were transferred onto 0.2 ?m nitrocellulose membranes and used for
Surface plasmon resonance experiments. Immobilization of antibodies
pling (Biacore) up to 8500–10,000 response units on CM5 sensor chips
(Biacore). Then, 60 ?l of analyte (A? oligomers) was injected on sensor
chips surfaces at a flow rate of 15 ?l/min and in various concentrations
(0.1–100 ?g/ml). PBS and a carbonate-bicarbonate buffer were used as
running buffers as specified. A? samples were kept at 4°C before each
chip regeneration was performed using 6.25–50 mM NaOH. Binding
Animals and drug treatment. We investigated female CRND8 trans-
genic mice carrying a double-mutated form of the human amyloid pre-
cursor protein 695 (APP695; the Swedish and Indiana mutations) under
the control of the Syrian hamster prion promoter, on a hybrid C57BL/
6–C3H/ HeJ background (courtesy of D. Westaway, University of To-
ronto, Toronto, Canada) (Chishti et al., 2001). Compared with other
murine models of AD (Ro ¨skam et al., 2010), TgCRND8 mice exhibit A?
plaques at a very young age (at ?3 months), accompanied by cognitive
deficits (Lovasic et al., 2005; Bacher et al., 2008). At 3 months (“young”;
n ? 15) or 12 months (“old”; n ? 15) of age, mice were injected intra-
experiments, the column FT was used as negative control (see above). In
addition, female APP23 transgenic mice, which express the human
APP751 isoform with the Swedish double mutation (K670N–M671L)
under control of a murine Thy1.2 expression cassette (courtesy of Dr.
trols and had been backcrossed with C57BL/6J mice for at least eight
generations (Sturchler-Pierrat et al., 1997). Mice were housed in groups
15 cm height) together with wild-type littermates with food and water
sections from FT- and NAbs–A?-treated young and old mice were stained for A? using anti-human A? monoclonal antibody 6F/3D. The average number (C, G) and area (D, H) of A?-
Dodeletal.•NaturallyOccurringAntibodiesandAlzheimer’sDiseaseJ.Neurosci.,April13,2011 • 31(15):5847–5854 • 5849
available ad libitum. All experimental procedures were in accordance
with the guidelines of the local animal care commission.
Brain tissue preparation. Mice were decapitated 5 d after the last injec-
tion. Brains were removed, and one hemisphere was immediately snap-
frozen in liquid nitrogen and kept at ?80°C until use. The other
dehydration and paraffin embedding.
sections of each transgenic animal were pretreated with formic acid and
incubated in a TechMate instrument (Dako) with 6F/3D (anti-A?
monoclonal antibody directed against residues 8–17; 1:100; Dako). The
second step used the Dako StreptABC complex horseradish peroxidase-
conjugated anti-mouse/rabbit antibody kit, and staining was developed
toxylin. The pairs of sections (10 ?m distances) were situated 100, 200,
were stained in five consecutive procedures, ensuring that the brains of
all three experimental groups were equally distributed.
Morphometry of the A? plaques. To quantify the A? plaque burden,
pus BX50, ColorView II, charge-coupled device camera; Olympus) un-
der constant light and filter settings. Color images were converted to
grayscale by extracting blue to gray values to obtain the best contrast
between positive immunoreactivity and the background. A constant
threshold was chosen for all images to detect immunoreactive staining
(analySIS 5; Soft Imaging System). The total number and size of plaques
were determined. Absolute values of plaque burden were related to the
Behavioral experiments. Fourteen- to 16-month-old Tg2576 mice ex-
pressing the Swedish mutation (K670N/M671L) of APP695 under the
control of the hamster prion promoter, on a hybrid C57BL/6–SJL back-
ground (Hsiao et al., 1996), were intraperitoneally injected with either
only (n ? 20). An additional group of age-matched wild-type mice were
treated with PBS (n ? 20).
To investigate the treatment efficacy of NAbs–A? on spatial memory,
we performed the object location test (Hale and Good, 2005). In brief,
which has been displaced to a new location. Animals that remember the
previous exposure spontaneously spend more time exploring the object
min trials with a 30 min intertrial delay. In trial 1, mice were allowed to
explore two similar objects located on one side of the cage for 10 min. In
trial 2, one of these objects was moved to a different location, and the
versus the total time spent exploring both objects in the test trial) was
calculated for each mouse. Object location test was reflected by more
time spent interacting with the object in the new location than with the
subject to the influence of individual variability in contact times with
objects and provides a measure of the extent of the discrimination be-
tween the novel and sample objects.
human antibody (MAHA) response, we treated 10 wild-type mice (C57
BL/6 genetic background of TgCRND8 mice used in drug treatment
experiments) with 200 ?g of NAbs–A? once a week for a period of 28 d
(Imboden et al., 2001). Blood was taken before and after the treatment
and investigated by MAHA ELISA. Briefly, plates were coated overnight
at 4°C with 5 ?g/ml human IgG1, diluted in sodium bicarbonate buffer,
plates were blocked for 3 h with SuperBlock (Thermo Fisher Scientific).
After washing four times with 1? PBS, 0.05% Tween 20, pH 7.4, the
serum samples were diluted 1:100 and added to the wells for overnight
incubation at 4°C. The wells were then washed four times with 1? PBS
and 0.05% Tween 20, pH 7.4, and then 0.1 ?g/ml HRP-conjugated goat
anti-mouse IgG was added and plates were incubated for 1 h at room
Tween 20, pH 7.4. A total of 100 ?l of tetramethylbenzidine (Calbio-
min, and then the reaction was stopped by the addition of 25 ?l/well of
The standard curve reagent used for this assay was obtained by perform-
ing these measurements on serial dilutions of mouse anti-human IgG1
(BD Biosciences Pharmingen) in concentrations between 1 and 1000
ng/ml in twofold increments.
Statistical analysis. Data were analyzed using the Student’s t test or
ANOVA as appropriate. All tests were applied (two tailed) using the
nificant at p ? 0.05.
induced cell death, we used A? preparations consisting of oli-
gomerized products (Kayed et al., 2003). NAbs–A? were able to
block the toxic effects of A? oligomers in SH-SY5Y cells in a
concentration-dependent manner (Fig. 1). Cell death was re-
duced by ?80% in the presence of NAbs–A?. Ig preparations
block the cytotoxic effects in this assay (Fig. 1).
Therapeutic efficacy of immunization has been associated
with the clearance of plaques in the brain of transgenic mice
(Schenk et al., 1999). Therefore, we investigated the ability of
mice (Chishti et al., 2001). We examined the effect of a low (80
?g) and high (200 ?g) dose of purified NAbs–A? on A? plaque
5850 • J.Neurosci.,April13,2011 • 31(15):5847–5854Dodeletal.•NaturallyOccurringAntibodiesandAlzheimer’sDisease
in young (4 months) and old (13 months) TgCRND8 mice. To
avoid the MAHA response, treatment with the foreign antibody
was limited to 4 weeks to prevent an adverse MAHA immune
response in the animal model. To investigate the MAHA re-
of ?0.01 mg/ml, and only one animal (0.06 mg/ml) had a titer
above 0.05 mg/ml. None of the animals died or exhibited clinical
or behavioral changes. None of the inspected organs showed
changes as investigated by pathological and histopathological
The cerebral plaque load in senescent
animals remained unaffected after 4
doses of NAbs–A? when compared with
age-matched sham-treated animals (Fig.
2A–D). In contrast, both low- and high
dose-treated young transgenic mice had a
significant reduction of plaque number
(high dose, by 24%, p ? 0.045; low dose,
by 34%, p ? 0.026) and plaque area (high
dose, by 47%, p ? 0.029; low dose, by
47%, p ? 0.028) (Fig. 2E–H). The differ-
ences between the high and the low doses
in young animals were not significant.
central and the peripheral pool of A? was
against A?; a phenomenon termed “pe-
ripheral sink hypothesis” (Dodart et al.,
2002; Dodel et al., 2002). Because the
NAbs–A? had no plaque-dissolving ben-
efit in the old animals, we further investi-
gated their effect on the concentration of
A? in CSF and plasma of immunized an-
of CSF A? in the animal group treated
with NAbs–A? (Fig. 2). In plasma, an in-
crease of A? was detectable, indicating an
periphery (Fig. 2J). In contrast, CSF and
plasma levels of A? did not change con-
siderably when treated with FT for the
Interestingly, NAbs–A? failed to de-
Similar results were
NAbs–A? was used to detect A? in the
brain vasculature of patients with cere-
bral amyloid angiopathy (Fig. 3D).
Antibodies raised against the A?
epitopes 4–10 have been reported to bind
to plaques in the brains of transgenic ani-
mals (McLaurin et al., 2002). Therefore,
we determined the linear sequences and
the conformational epitopes that were
recognized by NAbs–A? using a large set of synthetic peptides
the full-length A? but not to the albumin control. The epitope re-
of the A? sequence (Fig. 4A), whereas N-terminal fragments were
not recognized by NAbs–A?. The peptides with an altered amino
The relevant amino acids within the epitope were confirmed by an
et al., 2005), we found that NAbs–A? preferentially recognized
dimers and trimers and, to a lesser extent, monomeric forms of
of the NAbs–A? toward the C-terminal region. We identified sequence AA28–40(42) as the major binding site. N-terminal
without exchanged amino acids) were also investigated: A?1-40; A?1-42; A?1-12; A?1-15; A?10-28; A?20-38; A?20-31; A?25-40;
40;M35A;V40A; A?1-40;I32A;M35A; A?1-40;I32A;V40A; A?1-40;I32A; A?28-42;A30G; A?28-42;G33A; A?28-42;M35A; A?28-42;V39C; A?28-42;I32A;
oligomeric forms of A?, whereas NAbs–A? only interacted with dimeric and trimeric forms but not monomeric forms. C–E,
the monomeric form of A?. D, The on-rate of the monoclonal antibody MTA to A? was higher compared with the on-rate of
Dodeletal.•NaturallyOccurringAntibodiesandAlzheimer’sDiseaseJ.Neurosci.,April13,2011 • 31(15):5847–5854 • 5851
mid-terminal part of the A? sequence detected all three investi-
assemblies using SPR are shown in Figure 4C–E.
Furthermore, in behavioral experiments in the Tg2576 trans-
genic mice, the object location memory improved significantly
mance levels of wild-type control mice. Mice treated with PBS
regularly performed at chance levels (discrimination ratio: 0.5 ?
To the best of our knowledge, this is the first report to link natu-
tabolism of peptides that are toxic to humans. We found that
NAbs–A? were readily detectable in healthy individuals, inter-
acted with A?, and promoted its degradation. In epitope map-
ping, NAbs–A? detected the mid-/C-terminal epitope of A?,
to bind and clear plaques, but they were
able to improve behavioral changes in a
transgenic mice model. Therefore, the
question is, how may NAbs–A? work if
they can only recognize the mid-/C-end-
terminal part of A?? According to recent
solid-nuclear magnetic resonance spectros-
copy studies of A? oligomerization, the A?
residues 12–24 and 30–40 adopt ?-strand
conformations and form parallel [or anti-
parallel, according to a proposed model
(Petkova et al., 2005)] ?-sheets through in-
al., 2002). This process is followed by oli-
gomerization of the A? peptides, as shown
ization, the C-terminal part of the A? pep-
tide must be recognized for the antibody to
bind to it. If an antibody recognizes the
N-terminal part of the A? peptide, oli-
In the active immunization studies,
It is likely that N-terminal epitopes of the
A? peptide were predominantly exposed
and were available as binding sites. Ac-
primarily antibodies that recognized the
N-terminal epitope (AA4–10) of the A?
sequence (McLaurin et al., 2002). Fur-
thermore, A? peptide is deposited in a
fibrillar form in the plaques, and the
N-terminal part of the A? peptide is predominantly available at
the “outside” of the plaque as an “epitope” for N-terminal active
In contrast, NAbs–A?—as mainly directed to the mid-/C-
terminal epitope of A?—were in fact not capable of binding to
the N terminus of A?. This finding could also explain why our
NAbs–A? did not clear plaques.
Furthermore, we found that our NAbs–A? preferentially
(Britschgi et al., 2009). This may be the mechanisms why treat-
14- to 16-month-old Tg2576 mice treated with either NAbs–A? or PBS. NAbs–A?-
immunized Tg2576 mice but not PBS-treated Tg2576 mice showed a preference for ex-
ploring objects moved to a new location, indicated by a discrimination ratio above 0.5
Neuropsychological testing in Tg2576 mice. Mean discrimination ratios for
antibodies because only the N-terminal part was available for T-cell interaction. Accordingly, only N-terminal antibodies were
5852 • J.Neurosci.,April13,2011 • 31(15):5847–5854Dodeletal.•NaturallyOccurringAntibodiesandAlzheimer’sDisease
ment with NAbs–A? in young animals led to a reduction in
Interestingly, NAbs–A? failed to interact with other
aggregating/?-sheet promoting proteins, such as ?-synuclein
and prion proteins (data not shown) (Du et al., 2003). Because
naturally occurring antibodies do not bind to distinct protein
sequences but rather recognize specific patterns of conservative
mon conformational epitope among those proteins; our current
data, however, suggest that NAbs–A? display a rather narrow
epitopal recognition pattern.
A? as “disease causation of AD” (Shankar et al., 2008). This in-
the metabolism of A? peptides. However, targeting dissolved
plaques is not the primary goal because these plaques are inert
reservoirs (“garbage dump”) and they help to trap toxic oligom-
ers in the brain. Recent evidence from preclinical experimental
studies as well as data from clinical trials support this notion
(Holmes et al., 2008).
Although our data showed an efflux of central A? into the
periphery that favored the efflux hypothesis of A? clearance (pe-
ripheral sink) (DeMattos et al., 2002), it was noticeable that a
small amount of NAbs–A? was capable of crossing the blood–
brain barrier and penetrating into the brain, as was shown using
radiolabeled NAbs–A? (Bacher et al., 2009). In previous studies,
this small amount of autoantibodies available in the brain was
brain (Schenk et al., 1999; Bard et al., 2000; Hock et al., 2002).
Therefore, we cannot rule out that additional mechanisms, in-
cluding a central degradation of A? oligomers in the presence of
NAbs–A? by microglial cells, may occur in the brain.
tibody pool, have a physiological role and convey a protective
effect by inhibiting oligomerization of A? peptides and conse-
quently degrading A?. We further postulate that this protective
effect is primarily mediated through interactions with the mid-/
products of A?. Furthermore, the increase of NAbs–A? in the
serum probably disrupts the equilibrium between central and
peripheral A? pools, leading to a shift from the CNS to the
may play in the protection from AD comes from clinical studies
(Dodel et al., 2004; Relkin et al., 2009). We are fully aware that
imental results reported here, warrant a large clinical trial to
investigate the effects of IVIg and human NAbs–A? in patients
United States of America and results will be most likely available
Bacher M, Dodel R, Aljabari B, Keyvani K, Marambaud P, Kayed R, Glabe C,
Goertz N, Hoppmann A, Sachser N, Klotsche J, Schnell S, Lewejohann L,
Al-Abed Y (2008) CNI-1493 inhibits Abeta production, plaque forma-
tion, and cognitive deterioration in an animal model of Alzheimer’s dis-
ease. J Exp Med 205:1593–1599.
Bacher M, Depboylu C, Du Y, Noelker C, Oertel WH, Behr T, Henriksen G,
Behe M, Dodel R (2009) Peripheral and central biodistribution of
(111)In-labeled anti-beta-amyloid autoantibodies in a transgenic mouse
model of Alzheimer’s disease. Neurosci Lett 449:240–245.
Bard F, Cannon C, Barbour R, Burke RL, Games D, Grajeda H, Guido T, Hu
K, Huang J, Johnson-Wood K, Khan K, Kholodenko D, Lee M, Lieber-
burg I, Motter R, Nguyen M, Soriano F, Vasquez N, Weiss K, Welch B,
SeubertP,SchenkD,YednockT (2000) Peripherallyadministeredanti-
reduce pathology in a mouse model of Alzheimer disease. Nat Med
Britschgi M, Olin CE, Johns HT, Takeda-Uchimura Y, LeMieux MC, Ru-
fibach K, Rajadas J, Zhang H, Tomooka B, Robinson WH, Clark CM,
Fagan AM, Galasko DR, Holtzman DM, Jutel M, Kaye JA, Lemere CA,
T (2009) Neuroprotectivenaturalantibodiestoassembliesofamyloido-
genic peptides decrease with normal aging and advancing Alzheimer’s
disease. Proc Natl Acad Sci U S A 106:12145–12150.
Chishti MA, Yang DS, Janus C, Phinney AL, Horne P, Pearson J, Strome R,
Zuker N, Loukides J, French J, Turner S, Lozza G, Grilli M, Kunicki S,
Morissette C, Paquette J, Gervais F, Bergeron C, Fraser PE, Carlson GA,
George-HyslopPS,WestawayD (2001) Early-onsetamyloiddeposition
of amyloid precursor protein 695. J Biol Chem 276:21562–21570.
Ashe KH (2005) Natural oligomers of the amyloid-beta protein specifi-
cally disrupt cognitive function. Nat Neurosci 8:79–84.
DeMattos RB, Bales KR, Cummins DJ, Dodart JC, Paul SM, Holtzman DM
(2001) Peripheral anti-A beta antibody alters CNS and plasma A beta
clearance and decreases brain A beta burden in a mouse model of Alzhei-
mer’s disease. Proc Natl Acad Sci U S A 98:8850–8855.
DeMattos RB, Bales KR, Cummins DJ, Paul SM, Holtzman DM (2002)
Brain to plasma amyloid-beta efflux: a measure of brain amyloid burden
in a mouse model of Alzheimer’s disease. Science 295:2264–2267.
Dodart JC, Bales KR, Gannon KS, Greene SJ, DeMattos RB, Mathis C, De-
Long CA, Wu S, Wu X, Holtzman DM, Paul SM (2002) Immunization
mer’s disease model. Nat Neurosci 5:452–457.
Dodel R, Hampel H, Depboylu C, Lin S, Gao F, Schock S, Ja ¨ckel S, Wei X,
Buerger K, Ho ¨ft C, Hemmer B, Mo ¨ller HJ, Farlow M, Oertel WH, Som-
mer N, Du Y (2002) Human antibodies against amyloid beta peptide: a
potential treatment for Alzheimer’s disease. Ann Neurol 52:253–256.
Dodel R, Neff F, Noelker C, Pul R, Du Y, Bacher M, Oertel W (2010) Intra-
venous immunoglobulins as a treatment for Alzheimer’s disease: ratio-
nale and current evidence. Drugs 70:513–528.
Dodel RC, Du Y, Depboylu C, Hampel H, Fro ¨lich L, Haag A, Hemmeter U,
Paulsen S, Teipel SJ, Brettschneider S, Spottke A, No ¨lker C, Mo ¨ller HJ,
WeiX,FarlowM,SommerN,OertelWH (2004) Intravenousimmuno-
Alzheimer’s disease. J Neurol Neurosurg Psychiatry 75:1472–1474.
Du Y, Dodel R, Hampel H, Buerger K, Lin S, Eastwood B, Bales K, Gao F,
Moeller HJ, Oertel W, Farlow M, Paul S (2001) Reduced levels of amy-
loid beta-peptide antibody in Alzheimer disease. Neurology 57:801–805.
Du Y, Wei X, Dodel R, Sommer N, Hampel H, Gao F, Ma Z, Zhao L, Oertel
WH, Farlow M (2003) Human anti-beta-amyloid antibodies block
beta-amyloid fibril formation and prevent beta-amyloid-induced neuro-
toxicity. Brain 126:1935–1939.
Glabe CG (2008) Structural classification of toxic amyloid oligomers. J Biol
Hale G, Good M (2005) Impaired visuospatial recognition memory but
normal object novelty detection and relative familiarity judgments in
adult mice expressing the APPswe Alzheimer’s disease mutation. Behav
Hock C, Konietzko U, Papassotiropoulos A, Wollmer A, Streffer J, von Rotz
RC, Davey G, Moritz E, Nitsch RM (2002) Generation of antibodies
specific for beta-amyloid by vaccination of patients with Alzheimer dis-
ease. Nat Med 8:1270–1275.
Holmes C, Boche D, Wilkinson D, Yadegarfar G, Hopkins V, Bayer A, Jones
RW, Bullock R, Love S, Neal JW, Zotova E, Nicoll JA (2008) Long-term
effects of Abeta42 immunisation in Alzheimer’s disease: follow-up of a
randomised, placebo-controlled phase I trial. Lancet 372:216–223.
Hsiao K, Chapman P, Nilsen S, Eckman C, Harigaya Y, Younkin S, Yang F,
Cole G (1996) Correlative memory deficits, Abeta elevation, and amy-
loid plaques in transgenic mice. Science 274:99–102.
Dodeletal.•NaturallyOccurringAntibodiesandAlzheimer’sDisease J.Neurosci.,April13,2011 • 31(15):5847–5854 • 5853
Imboden M, Murphy KR, Rakhmilevich AL, Neal ZC, Xiang R, Reisfeld RA, Download full-text
Gillies SD, Sondel PM (2001) The level of MHC class I expression on
murine adenocarcinoma can change the antitumor effector mechanism
of immunocytokine therapy. Cancer Res 61:1500–1507.
Jia CY, Nie J, Wu C, Li C, Li SS (2005) Novel Src homology 3 domain-
Cell Proteomics 4:1155–1166.
KaetherC,SchmittS,WillemM,HaassC (2006) Amyloidprecursorprotein
and Notch intracellular domains are generated after transport of their
precursors to the cell surface. Traffic 7:408–415.
Kayed R, Head E, Thompson JL, McIntire TM, Milton SC, Cotman CW,
Glabe CG (2003) Common structure of soluble amyloid oligomers im-
plies common mechanism of pathogenesis. Science 300:486–489.
Khandogin J, Brooks CL 3rd (2007) Linking folding with aggregation in
Alzheimer’s beta-amyloid peptides. Proc Natl Acad Sci U S A 104:
Klyubin I, Walsh DM, Lemere CA, Cullen WK, Shankar GM, Betts V,
Spooner ET, Jiang L, Anwyl R, Selkoe DJ, Rowan MJ (2005) Amyloid
beta protein immunotherapy neutralizes Abeta oligomers that disrupt
synaptic plasticity in vivo. Nat Med 11:556–561.
Lesne ´ S,KohMT,KotilinekL,KayedR,GlabeCG,YangA,GallagherM,Ashe
KH (2006) A specific amyloid-beta protein assembly in the brain im-
pairs memory. Nature 440:352–357.
Lovasic L, Bauschke H, Janus C (2005) Working memory impairment in a
mer’s disease. Genes Brain Behav 4:197–208.
Mangialasche F, Solomon A, Winblad B, Mecocci P, Kivipelto M (2010)
Alzheimer’s disease: clinical trials and drug development. Lancet Neurol
Westaway D, Fraser PE, Mount HT, Przybylski M, St George-Hyslop P
target amyloid-beta residues 4–10 and inhibit cytotoxicity and fibrillo-
genesis. Nat Med 8:1263–1269.
Monsonego A, Maron R, Zota V, Selkoe DJ, Weiner HL (2001) Immune
hyporesponsiveness to amyloid beta-peptide in amyloid precursor pro-
tein transgenic mice: implications for the pathogenesis and treatment of
Alzheimer’s disease. Proc Natl Acad Sci U S A 98:10273–10278.
Morgan D, Diamond DM, Gottschall PE, Ugen KE, Dickey C, Hardy J, Duff
K, Jantzen P, DiCarlo G, Wilcock D, Connor K, Hatcher J, Hope C,
Gordon M, Arendash GW (2000) A beta peptide vaccination prevents
memory loss in an animal model of Alzheimer’s disease. Nature 408:
Petkova AT, Ishii Y, Balbach JJ, Antzutkin ON, Leapman RD, Delaglio F,
TyckoR (2002) AstructuralmodelforAlzheimer’sbeta-amyloidfibrils
based on experimental constraints from solid state NMR. Proc Natl Acad
Sci U S A 99:16742–16747.
Petkova AT, Leapman RD, Guo Z, Yau WM, Mattson MP, Tycko R (2005)
Self-propagating, molecular-level polymorphism in Alzheimer’s beta-
amyloid fibrils. Science 307:262–265.
(2006) Alzheimer’s disease beta-amyloid peptides are released in associ-
ation with exosomes. Proc Natl Acad Sci U S A 103:11172–11177.
Relkin NR, Szabo P, Adamiak B, Burgut T, Monthe C, Lent RW, Younkin S,
YounkinL,SchiffR,WekslerME (2009) 18-Monthstudyofintravenous
Ro ¨skam S, Neff F, Schwarting R, Bacher M, Dodel R (2010) APP transgenic
mice: the effect of active and passive immunotherapy in cognitive tasks.
Neurosci Biobehav Rev 34:487–499.
S, Wogulis M, Yednock T, Games D, Seubert P (1999) Immunization
with amyloid-beta attenuates Alzheimer-disease-like pathology in the
PDAPP mouse. Nature 400:173–177.
SelkoeDJ (2004) Cellbiologyofproteinmisfolding:theexamplesofAlzhei-
mer’s and Parkinson’s diseases. Nat Cell Biol 6:1054–1061.
Shankar GM, Li S, Mehta TH, Garcia-Munoz A, Shepardson NE, Smith I,
Brett FM, Farrell MA, Rowan MJ, Lemere CA, Regan CM, Walsh DM,
Sabatini BL, Selkoe DJ (2008) Amyloid-beta protein dimers isolated di-
rectly from Alzheimer’s brains impair synaptic plasticity and memory.
Nat Med 14:837–842.
Shoenfeld Y, Gershwin ME, Meroni PL (2007) Autoantibodies. Burlington:
Elsevier Science and Technology.
ME, Jucker M, Probst A, Staufenbiel M, Sommer B (1997) Two amyloid
precursor protein transgenic mouse models with Alzheimer disease-like
pathology. Proc Natl Acad Sci U S A 94:13287–13292.
RP, Ramsland PA, Edmundson AB, Weksler ME, Paul S (2008)
Autoantibody-catalyzed hydrolysis of amyloid beta peptide. J Biol Chem
Tomic JL, Pensalfini A, Head E, Glabe CG (2009) Soluble fibrillar oligomer
levels are elevated in Alzheimer’s disease brain and correlate with cogni-
tive dysfunction. Neurobiol Dis 35:352–358.
Townsend M, Shankar GM, Mehta T, Walsh DM, Selkoe DJ (2006) Effects
of secreted oligomers of amyloid beta-protein on hippocampal synaptic
plasticity: a potent role for trimers. J Physiol 572:477–492.
Walsh DM, Townsend M, Podlisny MB, Shankar GM, Fadeeva JV, El Agnaf
O, Hartley DM, Selkoe DJ (2005) Certain inhibitors of synthetic amy-
loid ?-peptide (A?) fibrillogenesis block oligomerization of natural A?
and thereby rescue long-term potentiation. J Neurosci 25:2455–2462.
Weksler ME, Relkin N, Turkenich R, LaRusse S, Zhou L, Szabo P (2002)
Patients with Alzheimer disease have lower levels of serum anti-amyloid
5854 • J.Neurosci.,April13,2011 • 31(15):5847–5854Dodeletal.•NaturallyOccurringAntibodiesandAlzheimer’sDisease