Soluble Oligomers Research Articles

Amyloid-β oligomers interact with NMDA receptors containing GluN2B subunits and metabotropic glutamate receptor 1 in primary cortical neurons: Relevance to the synapse pathology of Alzheimer’s disease

Recent evidence suggests that soluble amyloid-β oligomers (AβOs) act as a key factor in the pathogenetic mechanism of Alzheimer's disease (AD). AβOs induce neurotoxic and synaptotoxic effects probably through binding to certain receptors, however it remains unclarified which receptors are most critically involved. In addition, dysregulation in glutamatergic signaling is implicated in AD. In this study, we used a rat primary cortical neuron model to investigate AβO-induced aberrations of synaptic proteins and binding of extracellular AβOs to candidate receptors in the glutamatergic system. Immunocytochemical analyses showed that both presynaptic (SNAP-25, synapsin I) and postsynaptic (spinophilin, homer 1b/c) proteins appeared to aberrantly dislocate from synapses upon AβO treatment. Double immunofluorescence staining of AβO-treated neurons without permeabilization pretreatment revealed that extracellular AβOs exist over neuronal soma and neurites and clearly colocalized with GluN1 and GluN2B subunits of NMDA receptors and metabotropic glutamate receptor 1 (mGluR1), but not with NMDA GluN2A subunits and mGluR5. AβO treatment altered neither total protein levels nor intracellular localizations of these receptors. These results suggest that extracellular AβOs specifically bind to both NMDA receptors containing GluN2B subunits and mGluR1. It is likely that binding of AβOs to these receptors induces various pathological responses, consequently leading to synaptic disruptions. Our study thus highlights the important roles of GluN2B-containing NMDA receptors and mGluR1 receptors in the synapse pathology in AD.

Lysine 63-linked ubiquitination of tau oligomers contributes to the pathogenesis of Alzheimer’s disease

Ubiquitin-modified tau aggregates are abundantly found in human brains diagnosed with Alzheimer’s disease (AD) and other tauopathies. Soluble tau oligomers (TauO) are the most neurotoxic tau species that propagate pathology and elicit cognitive deficits, but whether ubiquitination contributes to tau formation and spreading is not fully understood. Here, we observed that K63-linked, but not K48-linked, ubiquitinated TauO accumulated at higher levels in AD brains compared with age-matched controls. Using mass spectrometry analyses, we identified 11 ubiquitinated sites on AD brain-derived TauO (AD TauO). We found that K63-linked TauO are associated with enhanced seeding activity and propagation in human tau-expressing primary neuronal and tau biosensor cells. Additionally, exposure of tau-inducible HEK cells to AD TauO with different ubiquitin linkages (wild type, K48, and K63) resulted in enhanced formation and secretion of K63-linked TauO, which was associated with impaired proteasome and lysosome functions. Multipathway analysis also revealed the involvement of K63-linked TauO in cell survival pathways, which are impaired in AD. Collectively, our study highlights the significance of selective TauO ubiquitination, which could influence tau aggregation, accumulation, and subsequent pathological propagation. The insights gained from this study hold great promise for targeted therapeutic intervention in AD and related tauopathies.

The Pathological G51D Mutation in Alpha-Synuclein Oligomers Confers Distinct Structural Attributes and Cellular Toxicity.

A wide variety of oligomeric structures are formed during the aggregation of proteins associated with neurodegenerative diseases. Such soluble oligomers are believed to be key toxic species in the related disorders; therefore, identification of the structural determinants of toxicity is of upmost importance. Here, we analysed toxic oligomers of α-synuclein and its pathological variants in order to identify structural features that could be related to toxicity and found a novel structural polymorphism within G51D oligomers. These G51D oligomers can adopt a variety of β-sheet-rich structures with differing degrees of α-helical content, and the helical structural content of these oligomers correlates with the level of induced cellular dysfunction in SH-SY5Y cells. This structure–function relationship observed in α-synuclein oligomers thus presents the α-helical structure as another potential structural determinant that may be linked with cellular toxicity in amyloid-related proteins.

Molecular Mechanisms of Amylin Turnover, Misfolding and Toxicity in the Pancreas.

Amyloidosis is a common pathological event in which proteins self-assemble into misfolded soluble and insoluble molecular forms, oligomers and fibrils that are often toxic to cells. Notably, aggregation-prone human islet amyloid polypeptide (hIAPP), or amylin, is a pancreatic hormone linked to islet β-cells demise in diabetics. The unifying mechanism by which amyloid proteins, including hIAPP, aggregate and kill cells is still matter of debate. The pathology of type-2 diabetes mellitus (T2DM) is characterized by extracellular and intracellular accumulation of toxic hIAPP species, soluble oligomers and insoluble fibrils in pancreatic human islets, eventually leading to loss of β-cell mass. This review focuses on molecular, biochemical and cell-biology studies exploring molecular mechanisms of hIAPP synthesis, trafficking and degradation in the pancreas. In addition to hIAPP turnover, the dynamics and the mechanisms of IAPP–membrane interactions; hIAPP aggregation and toxicity in vitro and in situ; and the regulatory role of diabetic factors, such as lipids and cholesterol, in these processes are also discussed.

Deducing the affinity between apolipoprotein E and soluble amyloid-beta oligomers using fluorescence correlation spectroscopy

Human apolipoprotein E (apoE), which plays a central role in lipid and cholesterol metabolism, exists in three major isoforms: E2, E3, and E4. The E4 allele is a major genetic factor for late-onset Alzheimer's disease (AD), while the E2 allele is thought to be protective. ApoE was shown to be connected to the aggregation, degradation, and clearance of toxic soluble oligomers of disordered amyloid-beta (Aβ) peptides, which are central to AD pathogenesis. However, previously reported interactions between apoE and Aβ are ambiguous since they rarely account for the dynamic and transient nature of Aβ oligomers or the aggregation and lipidation of apoE.

Effects of vesicular membrane composition on amyloid-beta oligomerization

Deposition of extracellular amyloid-β (Aβ) aggregates in the brain is one of the prime hallmarks in Alzheimer disease (AD). While high molecular weight Aβ plaques are observed in post-mortem AD brains, low-molecular weight, soluble Aβ oligomers are the primary toxic agents. Formation of these Aβ oligomers can be catalyzed by several interacting partners such as metal ions, other proteins, and lipids. We have previously demonstrated the generation of conformationally distinct oligomers of Aβ in the presence of lipid micelles.

Design of controllable degradable epoxy resin: High performance and feasible upcycling

AbstractEndowing epoxy resin with high performance, controllable degradation as well as feasible upcycling is a huge challenge. Herein, we synthesized a bio‐based amine curing agent containing spiro diacetal unit using an environment‐friendly method, then the controllable degradable epoxy resin was obtained after the curing reaction between diamine (DVPM) and bisphenol A epoxy resin (DGEBA). Results showed that DVPM‐DGEBA resin performed high glass translation temperature of 136°C, glassy storage modulus of 3.93 GPa, and tensile strength of 85.9 MPa, owing to spiro diacetal unit and abundant methoxyl‐related hydrogen bonds confirmed by non‐isothermal fourier transform infrared spectra (FT‐IR) and dynamic mechanical analyses. More importantly, DVPM‐DGEBA resin could decompose into soluble linear oligomers in mild acidic solution. Systematically investigated by 1H nuclear magnetic resonance, gel permeation chromatography, and FT‐IR. Furthermore, the recycled oligomers could be reprocessed into thermosetting resins in the presence of diamines, and the upcycling resin performed high‐performance (glass translation temperature of 170°C, glassy storage modulus of 6.03 GPa and tensile strength of 86.4 MPa) and flame‐retardant (V‐0 rating). This work gives us some new ideas to achieve the upcycling of the thermoset resin.

A Dual-Pronged Approach: A Ruthenium(III) Complex That Modulates Amyloid-β Aggregation and Disrupts Its Formed Aggregates.

Alzheimer's disease (AD) is a devastating neurological disorder for which soluble oligomers of the peptide amyloid-β (Aβ) are now recognized as the neurotoxic species. Metal-based therapeutics are uniquely suited to target Aβ, with ruthenium-based (Ru) complexes emerging as propitious candidates. Recently, azole-based Ru(III) complexes were observed to modulate the aggregation of Aβ in solution, where the inclusion of a primary amine proximal to the ligand coordination site improved the activity of the complexes. To advance these structure-activity relationships, a series of oxazole-based Ru complexes were prepared and evaluated for their ability to modulate Aβ aggregation. From these studies, a lead candidate, Oc, emerged that had superior activity relative to its azole predecessors in modulating the aggregation of soluble Aβ and diminishing its cytotoxicity. Further evaluation of Oc demonstrated its ability to disrupt formed Aβ aggregates, resulting in smaller amorphous species. Because altering both sides of the aggregation equilibrium for Aβ has not been previously suggested for metal-based complexes for AD, this work represents an exciting new avenue for improved therapeutic success.

A calcium-sensitive antibody isolates soluble amyloid-β aggregates and fibrils from Alzheimer's disease brain.

Aqueously soluble oligomers of amyloid-β peptide may be the principal neurotoxic forms of amyloid-β in Alzheimer’s disease, initiating downstream events that include tau hyperphosphorylation, neuritic/synaptic injury, microgliosis and neuron loss. Synthetic oligomeric amyloid-β has been studied extensively, but little is known about the biochemistry of natural oligomeric amyloid-β in human brain, even though it is more potent than simple synthetic peptides and comprises truncated and modified amyloid-β monomers. We hypothesized that monoclonal antibodies specific to neurotoxic oligomeric amyloid-β could be used to isolate it for further study.Here we report a unique human monoclonal antibody (B24) raised against synthetic oligomeric amyloid-β that potently prevents Alzheimer’s disease brain oligomeric amyloid-β-induced impairment of hippocampal long-term potentiation. B24 binds natural and synthetic oligomeric amyloid-β and a subset of amyloid plaques, but only in the presence of Ca2+. The amyloid-β N terminus is required for B24 binding. Hydroxyapatite chromatography revealed that natural oligomeric amyloid-β is highly avid for Ca2+. We took advantage of the reversible Ca2+-dependence of B24 binding to perform non-denaturing immunoaffinity isolation of oligomeric amyloid-β from Alzheimer’s disease brain-soluble extracts.Unexpectedly, the immunopurified material contained amyloid fibrils visualized by electron microscopy and amenable to further structural characterization. B24-purified human oligomeric amyloid-β inhibited mouse hippocampal long-term potentiation. These findings identify a calcium-dependent method for purifying bioactive brain oligomeric amyloid-β, at least some of which appears fibrillar.

The amyloid concentric β-barrel hypothesis: Models of amyloid beta 42 oligomers and annular protofibrils.

Amyloid beta (Aβ) peptides are a major contributor to Alzheimer's disease. They occur in differing lengths, each of which forms a multitude of assembly types. The most toxic soluble oligomers are formed by Aβ42; some of which have antiparallel β-sheets. Previously, our group proposed molecular models of Aβ42 hexamers in which the C-terminus third of the peptide (S3) forms an antiparallel 6-stranded β-barrel that is surrounded by an antiparallel barrel formed by the more polar N-terminus (S1) and middle (S2) portions. These hexamers were proposed to act as seeds from which dodecamers, octadecamers, both smooth annular protofibrils (sAPFs) and beaded annular protofibrils (bAPFs), and transmembrane channels form. Since then, numerous aspects of our models have been supported by experimental findings. Recently, NMR-based structures have been proposed for Aβ42 tetramers and octamers, and NMR studies have been reported for oligomers composed of ~32 monomers. Here we propose a range of concentric β-barrel models and compare their dimensions to image-averaged electron micrographs of both bAPFs and sAPFs of Aβ42. The smaller oligomers have 6, 8, 12, 16, and 18 monomers. These beads string together to form necklace-like bAPFs. These bAPRs gradually morph into sAPFs in which a S3 β-barrel is shielded on one or both sides by β-barrels formed from S1 and S2 segments.

Visualizing the Multistep Process of Protein Aggregation in Live Cells.

Protein aggregation is a biological phenomenon in which aberrantly processed or mutant proteins misfold and assemble into a variety of insoluble aggregates. Decades of studies have delineated the structure, interaction, and activity of proteins in either their natively folded structures or insoluble aggregates such as amyloid fibrils. However, a variety of intermediate species exist between these two extreme states in the protein folding landscape. Herein, we collectively term these intermediate species as misfolded protein oligomers, including soluble oligomers and preamyloid oligomers that are formed by unfolded or misfolded proteins. While extensive tools have been developed to study folded proteins or amyloid fibrils, research to understand the properties and activities of misfolded protein oligomers has been limited by the lack of methods to detect and interrogate these species in live cells.In this Account, we describe our efforts in the development of chemical methods that allow for the characterization of the multistep protein aggregation process, in particular the misfolded protein oligomers, in living cells. As the start of this journey, we attempted to develop a fluorogenic method wherein the misfolded oligomers could turn on the fluorescence of chemical probes that are conjugated to the protein-of-interest (POI). To this end, we produced a series of destabilized HaloTag variants, formulating the primary component of the AgHalo sensor, which misfolds and aggregates when cells are subjected to stress. When AgHalo is covalently conjugated with a solvatochromic fluorophore, misfolding of the AgHalo conjugate would activate fluorescence, resulting in the observation of misfolded oligomers. Following this work, we extended the scope of detection from AgHalo to any protein-of-interest via the AggTag method, wherein the POIs are genetically fused to self-labeling protein tags (HaloTag or SNAP-tag). Focusing on the molecular rotor-based fluorophores, we applied the modulated fluorescent protein (FP) chromophore core as a prototype for the AggTag probes, to enable the fluorogenic detection of misfolded soluble oligomers of multiple proteins in live cells. Next, we further developed the AggTag method to distinguish insoluble aggregates from misfolded oligomers, using two classes of probes that activate different fluorescence emission toward these two conformations. To enable this goal, we applied physical organic chemistry and computational chemistry to discover a new category of triode-like fluorophores, wherein the π orbitals of either an electron density regulator or the donor-acceptor linkages are used to control the rotational barriers of fluorophores in the excited states. This mechanism allows us to rationally design molecular rotor-based fluorophores that have desired responses to viscosity, thus extending the application of the AggTag method.In summary, our work allows the direct monitoring of the misfolded protein oligomers and differentiation of insoluble aggregates from other conformations in live cells, thus enabling studies of many currently unanswered questions in protein aggregation. Future directions are to develop methods that enable quantitative analyses of the protein aggregation process. Further, new methods are needed to detect and to quantify the formation and maturation of protein or RNA condensates that form membraneless organelles.

Understanding the pathophysiology of alzheimer's disease

Background: Neurodegenerative diseases, particularly Alzheimer's disease (AD), pose a significant health challenge globally, with projections indicating nearly 152 million affected individuals by 2050. AD accounts for 60% to 80% of neurodegenerative cases, manifesting primarily as sporadic Alzheimer's disease (SAD) after age 65. Aim: This review aims to elucidate the pathophysiology of AD, focusing on the early identification of biomarkers for diagnosis and the exploration of potential therapeutic interventions. Methods: A comprehensive literature review was conducted, examining the biological mechanisms underpinning AD, particularly the role of amyloid plaques and neurofibrillary tangles, along with the impact of lipid nutrients and nanotechnology in treatment delivery. Results: Key findings indicate that soluble amyloid-beta oligomers are critical in AD pathogenesis, contributing to synaptic dysfunction and cognitive decline. Moreover, recent advancements in nanotechnology, particularly through nanoliposomes, show promise for enhancing drug delivery across the blood-brain barrier. Conclusion: Understanding the complex interplay of genetic, environmental, and pathological factors in AD can inform early diagnostic strategies and therapeutic approaches. The role of lifestyle and dietary interventions is crucial, and future research should focus on leveraging nanotechnology for effective treatment delivery.

Amphiphilic stilbene derivatives attenuate the neurotoxicity of soluble Aβ42 oligomers by controlling their interactions with cell membranes.

The misfolded proteins or polypeptides commonly observed in neurodegenerative diseases, including Alzheimer's disease (AD), are promising drug targets for developing therapeutic agents. To target the amyloid-β (Aβ) peptide plaques and oligomers, the hallmarks of AD, we have developed twelve amphiphilic small molecules with different hydrophobic and hydrophilic fragments. In vitro fluorescence binding assays demonstrate that these amphiphilic compounds show high binding affinity to both Aβ plaques and oligomers, and six of them exhibit selective binding toward Aβ oligomers. These amphiphilic compounds can also label the Aβ species in the brain sections of transgenic AD mice, as shown by immunostaining with an Aβ antibody. Molecular docking studies were performed to obtain structure-affinity relationships. To our delight, four amphiphilic compounds can alleviate the Cu2+-Aβ induced toxicity in cell viability assays. In addition, confocal fluorescence imaging studies provide evidence that two compounds, ZY-15-MT and ZY-15-OMe, can disrupt the interactions between Aβ oligomers and human neuroblastoma SH-SY5Y cell membranes. Overall, these studies strongly suggest that developing compounds with amphiphilic properties that target Aβ oligomers and modulate the Aβ oligomer-cell membrane interactions can be an effective strategy for the development of small molecule AD therapeutics.

Early life stress-induced neuroinflammation and neurological disorders: a novel perspective for research.

Early life stress-induced neuroinflammation and neurological disorders: a novel perspective for research.

Molecular Dynamics Simulations of Protein Aggregation: Protocols for Simulation Setup and Analysis with Markov State Models and Transition Networks.

Protein disorder and aggregation play significant roles in the pathogenesis of numerous neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases. The end products of the aggregation process in these diseases are highly structured amyloid fibrils. Though in most cases, small, soluble oligomers formed during amyloid aggregation are the toxic species. A full understanding of the physicochemical forces that drive protein aggregation is thus required if one aims for the rational design of drugs targeting the formation of amyloid oligomers. Among a multitude of biophysical and biochemical techniques that are employed for studying protein aggregation, molecular dynamics (MD) simulations at the atomic level provide the highest temporal and spatial resolution of this process, capturing key steps during the formation of amyloid oligomers. Here we provide a step-by-step guide for setting up, running, and analyzing MD simulations of aggregating peptides using GROMACS. For the analysis, we provide the scripts that were developed in our lab, which allow to determine the oligomer size and inter-peptide contacts that drive the aggregation process. Moreover, we explain and provide the tools to derive Markov state models and transition networks from MD data of peptide aggregation.

A FRET-based aptasensor for the detection of α-synuclein oligomers as biomarkers of Parkinson's disease.

Soluble oligomers of α-synuclein (α-syn) are known to be responsible for neuronal death in the early stages of Parkinson's disease (PD). Thus, the development of a simple, rapid, and inexpensive method for the detection of α-syn oligomers can help PD diagnosis before irreversible damage to the brain tissue occurs. The present study is aimed at developing a FRET-based aptasensor for the selective and sensitive detection of α-syn oligomers. Herein, FAM-labeled aptamer adsorption on graphene oxide (GO) resulted in fluorescence quenching of FAM. The binding of α-syn oligomers to the aptamer led to the recovery of the fluorescence intensity. Under optimized conditions, the developed biosensor showed two linear response ranges from 10-100 nM and 250 nM to 2 μM in α-syn oligomer detection. The limit of detection (LOD) and limit of quantification (LOQ) were calculated to be 6.3 nM and 19.3 nM, respectively. Furthermore, the performance of the sensing platform in the detection of the target analyte in biological matrices was demonstrated by the assay of α-syn oligomers in spiked human saliva samples. According to the obtained results, this sensing platform has a good performance for α-syn oligomer detection and it can be considered as a promising candidate for the early diagnosis of PD.

Structural determinants of the interactions of catechins with Aβ oligomers and lipid membranes

The aberrant self-assembly of intrinsically disordered proteins (IDPs) into soluble oligomers and their interactions with biological membranes underlie the pathogenesis of numerous neurodegenerative diseases, including Alzheimer’s disease. Catechins have emerged as useful tools to reduce the toxicity of IDP oligomers by modulating their interactions with membranes. However, the structural determinants of catechin binding to IDP oligomers and membranes remain largely elusive. Here, we assemble a catechin library by combining several naturally occurring chemical modifications and, using a coupled NMR-statistical approach, we map at atomic resolution the interactions of such library with the Alzheimer’s-associated amyloid-beta (Aβ) oligomers and model membranes. Our results reveal multiple catechin affinity drivers and show that the combination of affinity-reducing covalent changes may lead to unexpected net gains in affinity. Interestingly, we find that the positive cooperativity is more prevalent for Aβ oligomers than membrane binding, and that the determinants underlying catechin recognition by membranes are markedly different from those dissected for Aβ oligomers. Notably, we find that the unanticipated positive cooperativity arises from the critical regulatory role of the gallate catechin moiety, which recruits previously disengaged substituents into the binding interface and leads to an overall greater compaction of the receptor-bound conformation. Overall, the previously elusive structural attributes mapped here provide an unprecedented foundation to establish structure-activity relationships of catechins.

Mechanisms Underlying the Antidiabetic Activities of Polyphenolic Compounds: A Review.

Polyphenolic compounds are thought to show considerable promise for the treatment of various metabolic disorders, including type 2 diabetes mellitus (T2DM). This review addresses evidence from in vitro, in vivo, and clinical studies for the antidiabetic effects of certain polyphenolic compounds. We focus on the role of cytotoxic human amylin (hA) aggregates in the pathogenesis of T2DM, and how polyphenols can ameliorate this process by suppressing or modifying their formation. Small, soluble amylin oligomers elicit cytotoxicity in pancreatic islet β-cells and may thus cause β-cell disruption in T2DM. Amylin oligomers may also contribute to oxidative stress and inflammation that lead to the triggering of β-cell apoptosis. Polyphenols may exert antidiabetic effects via their ability to inhibit hA aggregation, and to modulate oxidative stress, inflammation, and other pathways that are β-cell-protective or insulin-sensitizing. There is evidence that their ability to inhibit and destabilize self-assembly by hA requires aromatic molecular structures that bind to misfolding monomers or oligomers, coupled with adjacent hydroxyl groups present on single phenyl rings. Thus, these multifunctional compounds have the potential to be effective against the pleiotropic mechanisms of T2DM. However, substantial further research will be required before it can be determined whether a polyphenol-based molecular entity can be used as a therapeutic for type 2 diabetes.

Electrostatics Drive Oligomerization and Aggregation of Human Interferon Alpha-2a.

Aggregation is a common phenomenon in the field of protein therapeutics and can lead to function loss or immunogenic patient responses. Two strategies are currently used to reduce aggregation: (1) finding a suitable formulation, which is labor-intensive and requires large protein quantities, or (2) engineering the protein, which requires extensive knowledge about the protein aggregation pathway. We present a biophysical characterization of the oligomerization and aggregation processes by Interferon alpha-2a (IFNα-2a), a protein drug with antiviral and immunomodulatory properties. This study combines experimental high throughput screening with detailed investigations by small-angle X-ray scattering and analytical ultracentrifugation. Metropolis Monte Carlo simulations are used to gain insight into the underlying intermolecular interactions. IFNα-2a forms soluble oligomers that are controlled by a fast pH and concentration-dependent equilibrium. Close to the isoelectric point of 6, IFNα-2a forms insoluble aggregates which can be prevented by adding salt. We show that monomer attraction is driven mainly by molecular anisotropic dipole–dipole interactions that increase with increasing pH. Repulsion is due to monopole–monopole interactions and depends on the charge of IFNα-2a. The study highlights how combining multiple methods helps to systematically dissect the molecular mechanisms driving oligomer formation and to design ultimately efficient strategies for preventing detrimental protein aggregation.

Development of a dual Aβ‐tau vaccine for the prevention of Alzheimer’s disease

AbstractBackgroundAlzheimer’s disease (AD) is characterized by two pathological hallmarks, Aβ plaques and tau tangles. Several antibodies targeting Aβ plaques showed signs of slowing clinical decline in AD. Antibodies targeting tau epitopes and vaccines targeting either Aβ or tau are in earlier clinical development stages. Since Aβ and tau may act synergistically in the cause and/or progression of disease, targeting both concomitantly may lead to better efficacy for prevention of AD. Here we describe the development of our dual Aβ‐tau vaccine for AD aimed to clear pathogenic Aβ and block tau transmission.MethodA variety of linear immunogens containing both Aβ and tau peptides separated by various linkers and dendritic cell cleavage sites were screened in mice and guinea pigs for the generation of balanced titers to Aβ and tau. A subset was also evaluated in cynomolgus monkey. Animals were immunized intramuscularly with peptide‐conjugated immunogens formulated with an appropriate adjuvant. Bleeds were taken 1‐2 week after each injection. Serum titers were determined against Aβ and tau. Fresh frozen human AD or control brain sections were stained with sera from immunized and control animals. The neutralizing activity of immune sera was also assessed against soluble Ab oligomers. T‐cell reactivity was also investigated.ResultWe found dual vaccine constructs that provided balanced and immunogenic responses against Aβ and tau proteins. Immune sera bound avidly to Aβ plaques and tau tangles in human AD brain sections at concentrations expected to be achieved in CNS under in vivo conditions. Immune sera also inhibited the binding of soluble Aβ aggregates to hippocampal neurons. Immunogens did not elicit cytotoxic T‐cell responses to Aβ or tau.ConclusionWe developed dual vaccine constructs that induce optimally balanced titers against Aβ and tau in cynomolgus monkeys. Antibodies were immunoreactive to both Aβ plaques and neurofibrillary tau tangles in human AD brain sections and blocked the binding of soluble Aβ aggregates to neurons, without eliciting T‐cell responses to Aβor tau. These results support the development of a dual Aβ‐tau vaccine with the ability to target pathogenic Aβ and tau for the prevention of AD.