NR AUTD
AU Zuber,C.; Rey,C.; Knackmuss,S.; Reusch,U.; Mitteregger,G.; Pace,C.; Fröhlich,T.; Arnold,G.J.; Büning,H.; Little,M.; Hallek,M.; Weiss,S.
TI Single chain antibodies against the prion protein receptor as a tool in TSE therapies
QU TSE-Forum, 6. Kongress - Nationale TSE-Forschungsplattform, Greifswald 26.6.-28.6.2006, Poster: Therapie THE-09
PT Konferenz-Poster
AB
Recently, we identified the 37/67 kDa laminin receptor (LRP/LR) as the cell surface receptor for the cellular prion protein (PrPc) (1) and as a receptor for BSE prions on human enterocytes (2). We proved that polyclonal LRP/LR specific antibodies are able to abolish PrPsc propagation in scrapie infected neuroblastoma cells (3), demonstrating that the disruption of the LRP-PrP interaction is a relevant strategy to treat prion diseases. In the aim to develop anti-TSE therapies based on antibodies, we performed a screen of single chain antibodies (scFv) on GST::LRP by phage display. Two scFvs termed N3 and S18 have been selected. We report that both scFvs recognize specifically the denatured form of LRP as well as the native form on the cell surface in BHK cells overexpressing LRP. We identified the epitopes on LRP recognized by S18 and N3, respectively, by an epitope mapping approach. The ability of both antibodies to interfere with the LRP-PrP interaction was proven by a pull-down assay. A potential therapeutic effect of the scFv S18 in a scrapie mouse model by passive immunization has been investigated. Although the S18 antibody reduced PrPsc levels in the spleen 90 days post i.p. scFv injection, incubation times have not been prolonged significantly. In order to deliver the scFvs directly into the brain by gene transfer, we injected recombinant scFv expressing Adeno-Associated-Viruses (AAV) stereotactically into the brain. In order to get a permanent release of the antibodies over a long period of time, we generated scFv secreting myotubes for grafting into mouse brains. Targeting LRP/LR by (i) passive immunization, (ii) gene therapy with recombinant AAVs and (iii) cell therapy employing grafting of scFv secreting muscle cells might represent alternative therapeutic approaches for the treatment of TSEs.
(1) Gauczynski et al. (2001) EMBO J. 20, 5863-5875. (2) Morel et al. (2005) Am J Path, 167, 1033-1042. (3) Leucht et al. (2003) EMBO rep 4, 290-295.
AD C.Zuber, C.Rey, S.Weiss, Laboratorium für Molekulare Biologie-Genzentrum-Institut für Biochemie der LMU München, Feodor-Lynen-Strasse 25, D-81377 Munich, Germany; S.Knackmuss, U.Reusch, M.Little, Affimed Therapeutics AG-Technologiepark- Im Neuheimer Feld 582, 69120 Heidelberg, Germany; H.Büning3, M.Hallek, Universität zu Köln, Klinik I für Innere Medizin, Joseph-Stelzmann-Str. 9, 50924 Köln, Germany; C.Pace, G.Mitteregger, Zentrum für Neuropathologie und Prionforschung der LMU München, Feodor-Lynen-Str. 23, 81377 München, Germany
SP englisch
PO Deutschland
EA Photo des in Greifswald gezeigten Posters, dessen Autorenliste nach Auskunft vn Chantal Zuber aktueller als der Tagungsband ist, aber noch um Hallek,M. ergänzt werden sollte, oben, unten
OR Tagungsband