Session Information
Date: Monday, October 8, 2018
Session Title: Parkinson's Disease: Pathophysiology
Session Time: 1:15pm-2:45pm
Location: Hall 3FG
Objective: We tested naturally occurring antibodies (nAbs) against α-synuclein both in vitro as well as in HEK293T cells to determine binding, cell viability and capacity to reduce αSyn aggregation.
Background: Parkinson disease (PD) is the second frequent neurodegenerative disorder and causes severe motor disturbances. nAbs are part of the innate immune system. They are produced without contact to the specific antigen they recognize and are polyclonal. Variation in specific nAbs levels are described for αSyn and tau. PD patients have lower levels of αSyn specific nAbs. nAbs-αSyn are thought to decrease the load of αSyn deposits in the brain, but the mechanism of action is fairly unclear. Therefore we want to elucidate the behavior of nAbs-αSyn in cell culture systems.
Methods: We isolated nAbs-αSyn from intravenous Immunoglobulins (IVIG) using column affinity purification. Recombinant human αSyn was coupled to AminoLink Plus coupling resin. IVIGs were incubated in the column repeatedly followed by a pH dependent elution step. nAbs-αSyn from several purification rounds were pooled to reach reasonable protein concentrations. We tested the pooled antibodies to both in vitro as well as in HEK293T cells.
Results: nAbs-αSyn could be isolated from human IVIGs. They bound monomeric αSyn in a dot blot as well as in an ELISA against aSyn. nAbs-αSyn did not affect cell viability as tested in a Cell Titer Blue and an LDH Assay. In an in vitro Thioflavin T aggregation assay they inhibited fibrillation of αSyn. In a cell culture system of HEK293T cells transfected with EGFP-tagged αSyn we saw a reduction of aggresome load as visible by fluorescence microscopy.
Conclusions: nAbs-αSyn inhibited aggregation of αSyn both in vitro and in cell culture systems. These findings have to be corroborated further. Next, we want to analyze the effects of nAbs-αSyn in a Parkinson mouse modell and describe the binding behavior of nAbs-αSyn to different conformational αSyn forms. As an outlook, we want to understand the mechanism of action of nAbs-αSyn to better extent and hopefully add antibodies to improved treatment of PD patients.
To cite this abstract in AMA style:
A. Braczynski, E. Agerschou, Y. Kronimus, W. Hoyer, R. Dodel, B. Falkenburger, J. Schulz, J. Bach. Natural occurring antibodies reduce aggregation of α-synuclein [abstract]. Mov Disord. 2018; 33 (suppl 2). https://www.mdsabstracts.org/abstract/natural-occurring-antibodies-reduce-aggregation-of-%ce%b1-synuclein/. Accessed June 15, 2025.« Back to 2018 International Congress
MDS Abstracts - https://www.mdsabstracts.org/abstract/natural-occurring-antibodies-reduce-aggregation-of-%ce%b1-synuclein/