Figure 1

Titers, kinetics and reactivity of AFF 1-induced antibodies after repeated immunization in MBP-α-syn tg mice. (A) Titers of antibodies against human and murine α-syn elicited after immunization with vehicle, original C-terminal α-syn antigen or AFF 1 (B) IgG response towards the immunizing peptide AFF 1 (as BSA conjugate) as well as against recombinant human α-syn and β-syn from plasma taken at end point (C) Kinetics of the IgG responses to the immunizing peptide following vaccination with vehicle or AFF 1 (D) Kinetics of the IgG responses to recombinant human α-syn following vaccination with vehicle or AFF 1 (E) α-syn immune reactivity of AFF 1-induced antibodies in brain sections of naïve non-tg, mThy1-α-syn tg and MBP-α-syn tg mice. Plasma from AFF 1-immunized MBP-α-syn tg mice was used at a final dilution of 1:100. As positive control, the human α-syn-specific antibody LB509 was used. An anti-mouse IgG antibody was used as negative control. Cell nuclei were stained with DAPI (blue) (F) Immunoblot analysis of AFF 1-induced antibodies against aggregated (o) and monomeric (m) recombinant α-syn. The human α-syn-specific antibody LB509 was used as positive control. Aggregated (oligomeric) α-syn was obtained by 4-hydroxy-2-nonenal treatment. Titers are depicted as OD max/2 at 405 nm. PP, Preplasma; EP, end point plasma; P1-P5, plasma taken after each immunization. For titer calculations, n = 10 animals per group.