title: The toxicity of antiprion antibodies is mediated by the flexible tail of the prion protein.
abstract: Prion infections cause lethal neurodegeneration. This process requires the cellular prion protein (PrP(C); ref. 1), which contains a globular domain hinged to a long amino-proximal flexible tail. Here we describe rapid neurotoxicity in mice and cerebellar organotypic cultured slices exposed to ligands targeting the α1 and α3 helices of the PrP(C) globular domain. Ligands included seven distinct monoclonal antibodies, monovalent Fab1 fragments and recombinant single-chain variable fragment miniantibodies. Similar to prion infections, the toxicity of globular domain ligands required neuronal PrP(C), was exacerbated by PrP(C) overexpression, was associated with calpain activation and was antagonized by calpain inhibitors. Neurodegeneration was accompanied by a burst of reactive oxygen species, and was suppressed by antioxidants. Furthermore, genetic ablation of the superoxide-producing enzyme NOX2 (also known as CYBB) protected mice from globular domain ligand toxicity. We also found that neurotoxicity was prevented by deletions of the octapeptide repeats within the flexible tail. These deletions did not appreciably compromise globular domain antibody binding, suggesting that the flexible tail is required to transmit toxic signals that originate from the globular domain and trigger oxidative stress and calpain activation. Supporting this view, various octapeptide ligands were not only innocuous to both cerebellar organotypic cultured slices and mice, but also prevented the toxicity of globular domain ligands while not interfering with their binding. We conclude that PrP(C) consists of two functionally distinct modules, with the globular domain and the flexible tail exerting regulatory and executive functions, respectively. Octapeptide ligands also prolonged the life of mice expressing the toxic PrP(C) mutant, PrP(Δ94-134), indicating that the flexible tail mediates toxicity in two distinct PrP(C)-related conditions. Flexible tail-mediated toxicity may conceivably play a role in further prion pathologies, such as familial Creutzfeldt-Jakob disease in humans bearing supernumerary octapeptides.
authors.affiliation: Institute of Neuropathology, University Hospital Zurich, Schmelzbergstrasse 12, CH-8091 Zurich, Switzerland.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Sonati
authors.forename: Regina R
authors.initials: RR
authors.lastname: Reimann
authors.forename: Jeppe
authors.initials: J
authors.lastname: Falsig
authors.forename: Pravas Kumar
authors.initials: PK
authors.lastname: Baral
authors.forename: Tracy
authors.initials: T
authors.lastname: O'Connor
authors.forename: Simone
authors.initials: S
authors.lastname: Hornemann
authors.forename: Sine
authors.initials: S
authors.lastname: Yaganoglu
authors.forename: Bei
authors.initials: B
authors.lastname: Li
authors.forename: Uli S
authors.initials: US
authors.lastname: Herrmann
authors.forename: Barbara
authors.initials: B
authors.lastname: Wieland
authors.forename: Mridula
authors.initials: M
authors.lastname: Swayampakula
authors.forename: Muhammad Hafizur
authors.initials: MH
authors.lastname: Rahman
authors.forename: Dipankar
authors.initials: D
authors.lastname: Das
authors.forename: Nat
authors.initials: N
authors.lastname: Kav
authors.forename: Roland
authors.initials: R
authors.lastname: Riek
authors.forename: Pawel P
authors.initials: PP
authors.lastname: Liberski
authors.forename: Michael N G
authors.initials: MN
authors.lastname: James
authors.forename: Adriano
authors.initials: A
authors.lastname: Aguzzi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000633:toxicity*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000633:toxicity; D001666:Binding Sites, Antibody; D002154:Calpain / Q000378:metabolism; D002531:Cerebellum; D007562:Creutzfeldt-Jakob Syndrome / Q000378:metabolism; D003432:Cross-Linking Reagents; D018604:Epitope Mapping; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000633:toxicity; D066298:In Vitro Techniques; D008024:Ligands; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism; D019636:Neurodegenerative Diseases / Q000378:metabolism; D018384:Oxidative Stress; D018583:Pliability; D018621:PrPC Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011328:Prions / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics; D057127:Single-Chain Antibodies / Q000276:immunology / Q000633:toxicity
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D007140:Immunoglobulin Fab Fragments; D008024:Ligands; D008562:Membrane Glycoproteins; D018621:PrPC Proteins; D011328:Prions; D017382:Reactive Oxygen Species; D057127:Single-Chain Antibodies; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D002154:Calpain
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