Novel monoclonal antibodies targeting the microtubule-binding domain of human tau

Croft, Cara L.; Moore, Brenda D.; Ran, Yong; Chakrabarty, Paramita; Levites, Yona; Golde, Todd E.

doi:10.1371/journal.pone.0195211

Cited by 16 publications

(15 citation statements)

References 47 publications

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“…BSCs for assessment of insoluble tau from three wells (nine slices in total) were harvested into ice-cold phosphate buffered saline (PBS) and presented as n = 1. Sarkosyl-insoluble extractions were performed as previously described [ 10 , 13 ]. In brief, the supernatant (S1) was kept and run on Western blots as the soluble fraction and the pellet (P3) was resuspended in 4 M Urea 2% SDS buffer and run on Western blots as the sarkosyl-insoluble fraction.…”

Section: Methodsmentioning

confidence: 99%

Photodynamic studies reveal rapid formation and appreciable turnover of tau inclusions

et al. 2021

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Accumulation of the tau protein in fibrillar intracellular aggregates is a defining feature of multiple neurodegenerative diseases collectively referred to as tauopathies. Despite intensive study of tau, there is limited information on the formation and clearance dynamics of tau inclusions. Using rAAV vectors to mediate expression of Dendra2-tagged human wild-type, P301L and pro-aggregant P301L/S320F tau proteins, with and without the addition of exogenous tau fibrillar seeds, we evaluated tau inclusion dynamics in organotypic brain slice culture (BSC) models using long-term optical pulse labeling methodology. Our studies reveal that tau inclusions typically form in 12–96 h in tauopathy BSC models. Unexpectedly, we demonstrate appreciable turnover of tau within inclusions with an average half-life of ~ 1 week when inclusions are newly formed. When BSCs with inclusions are aged in culture for extended periods, tau inclusions continue to turnover, but their half-lives increase to ~ 2 weeks and ~ 3 weeks after 1 and 2 months in culture, respectively. Individual tau inclusions can be long-lived structures that can persist for months in these BSC models and for even longer in the human brain. However, our data indicate that tau inclusions, are not ‘tombstones’, but dynamic structures with appreciable turnover. Understanding the cellular processes mediating this inclusion turnover may lead to new therapeutic strategies that could reverse pathological tau inclusion formation.

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Section: Methodsmentioning

confidence: 99%

Photodynamic studies reveal rapid formation and appreciable turnover of tau inclusions

et al. 2021

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“…BALC/c mice (Jackson Laboratory, Bar Harbor, ME) were immunized with a synthetic peptide Cys-202 SPGTPGpSRSRTP 213 (synthesized and purified by GenScript USA Inc., Piscataway, NJ), which corresponds to tau phosphorylated at Ser208, conjugated to KLH as previously described [16,73,74]. Hybridoma clones were screened for their specificity by ELISA [16,73]. Monoclonal antibody 3G12 was found to be specific for tau phosphorylated at Ser208 by ELISA and was useful for immunohistochemistry and western blotting.…”

Section: Generation Of Monoclonal Antibody To Tau Phosphorylated At Smentioning

confidence: 99%

Tau Ser208 phosphorylation promotes aggregation and reveals neuropathologic diversity in Alzheimer’s disease and other tauopathies

Xia

Prokop

Gorion

et al. 2020

acta neuropathol commun

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Tau protein abnormally aggregates in tauopathies, a diverse group of neurologic diseases that includes Alzheimer's disease (AD). In early stages of disease, tau becomes hyperphosphorylated and mislocalized, which can contribute to its aggregation and toxicity. We demonstrate that tau phosphorylation at Ser208 (pSer208) promotes microtubule dysfunction and tau aggregation in cultured cells. Comparative assessment of the epitopes recognized by antibodies AT8, CP13, and 7F2 demonstrates that CP13 and 7F2 are specific for tau phosphorylation at Ser202 and Thr205, respectively, independently of the phosphorylation state of adjacent phosphorylation sites. Supporting the involvement of pSer208 in tau pathology, a novel monoclonal antibody 3G12 specific for tau phosphorylation at Ser208 revealed strong reactivity of tau inclusions in the brains of PS19 and rTg4510 transgenic mouse models of tauopathy. 3G12 also labelled neurofibrillary tangles in brains of patients with AD but revealed differential staining compared to CP13 and 7F2 for other types of tau pathologies such as in neuropil threads and neuritic plaques in AD, tufted astrocytes in progressive supranuclear palsy and astrocytic plaques in corticobasal degeneration. These results support the hypothesis that tau phosphorylation at Ser208 strongly contributes to unique types of tau aggregation and may be a reliable marker for the presence of mature neurofibrillary tangles.

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“…H150 was raised against amino acids 1-150 of human tau, while 3026 was raised against full length recombinant 0N3R human tau. 81A11 is a mouse monoclonal antibody specific for the R2 region of tau [34]. Mouse monoclonal 2G2 was generated by immunizing BALB/c mice with phosphopeptide CKHPGGGpSVQIVYKPVDL synthesized and purified by GenScript USA Inc (Piscataway, NJ) and conjugated to imject maleimideactivated mariculture keyhole limpet hemocyanin (mcKLH; Thermo Scientific, Waltham, MA).…”

Section: Antibodiesmentioning

confidence: 99%

Phosphorylation of serine 305 in tau inhibits aggregation

Strang

Sorrentino

Riffe

et al. 2019

Neuroscience Letters

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Alzheimer's disease and other tauopathies are characterized by the brain accumulation of hyperphosphorylated aggregated tau protein forming pathological inclusions. Although elevated tau phosphorylated at many amino acid residues is a hallmark of pathological tau, some evidence suggest that tau phosphorylation at unique sites, especially within its microtubule-binding domain, might inhibit aggregation. In this study, the effects of phosphorylation of two unique residues within this domain, serine 305 (S305) and serine 320 (S320), were examined in the context of established aggregation and seeding models. It was found that the S305E phosphomimetic significantly inhibited both tau seeding and tau aggregation in this model, while S320E did not. To further explore S305 phosphorylation in vivo, a monoclonal antibody (2G2) specific for tau phosphorylated at S305 was generated and characterized. Consistent with inhibition of tau aggregation, phosphorylation of S305 was not detected in pathological tau inclusions in Alzheimer's disease brain tissue. This study indicates that phosphorylation of unique tau residues can be inhibitory to aggregate formation, and has important implications for potential kinase therapies. Additionally, it creates new tools for observing these changes in vivo.

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Novel monoclonal antibodies targeting the microtubule-binding domain of human tau

Cited by 16 publications

References 47 publications

Photodynamic studies reveal rapid formation and appreciable turnover of tau inclusions

Photodynamic studies reveal rapid formation and appreciable turnover of tau inclusions

Tau Ser208 phosphorylation promotes aggregation and reveals neuropathologic diversity in Alzheimer’s disease and other tauopathies

Phosphorylation of serine 305 in tau inhibits aggregation

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