title: Antibodies against alpha-synuclein reduce oligomerization in living cells.
abstract: Recent research implicates soluble aggregated forms of α-synuclein as neurotoxic species with a central role in the pathogenesis of Parkinson's disease and related disorders. The pathway by which α-synuclein aggregates is believed to follow a step-wise pattern, in which dimers and smaller oligomers are initially formed. Here, we used H4 neuroglioma cells expressing α-synuclein fused to hemi:GFP constructs to study the effects of α-synuclein monoclonal antibodies on the early stages of aggregation, as quantified by Bimolecular Fluorescence Complementation assay. Widefield and confocal microscopy revealed that cells treated for 48 h with monoclonal antibodies internalized antibodies to various degrees. C-terminal and oligomer-selective α-synuclein antibodies reduced the extent of α-synuclein dimerization/oligomerization, as indicated by decreased GFP fluorescence signal. Furthermore, ELISA measurements on lysates and conditioned media from antibody treated cells displayed lower α-synuclein levels compared to untreated cells, suggesting increased protein turnover. Taken together, our results propose that extracellular administration of monoclonal antibodies can modify or inhibit early steps in the aggregation process of α-synuclein, thus providing further support for passive immunization against diseases with α-synuclein pathology.
authors.affiliation: Rudbeck Laboratory, Department of Public Health/Geriatrics, Uppsala University, Uppsala, Sweden.
authors.forename: Thomas
authors.initials: T
authors.lastname: Näsström
authors.forename: Susana
authors.initials: S
authors.lastname: Gonçalves
authors.forename: Charlotte
authors.initials: C
authors.lastname: Sahlin
authors.forename: Eva
authors.initials: E
authors.lastname: Nordström
authors.forename: Valentina
authors.initials: V
authors.lastname: Screpanti Sundquist
authors.forename: Lars
authors.initials: L
authors.lastname: Lannfelt
authors.forename: Joakim
authors.initials: J
authors.lastname: Bergström
authors.forename: Tiago F
authors.initials: TF
authors.lastname: Outeiro
authors.forename: Martin
authors.initials: M
authors.lastname: Ingelsson
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D015153:Blotting, Western; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D005910:Glioma / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D055503:Protein Multimerization / Q000187:drug effects* / Q000276:immunology; D014407:Tumor Cells, Cultured; D051844:alpha-Synuclein / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D051844:alpha-Synuclein; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
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