NR AWRC
AU Zuber,C.; Rey,C.; Knackmuss,S.; Reusch,U.; Pace,C.; Mitteregger,G.; Little,M.; Weiss,S.
TI Passive immunotransfer of antibodies directed against the prion protein receptor lrp/lr as a therapeutic strategy in TSE therapy
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions THE-23
PT Konferenz-Poster
AB
Recently we identified the 37/67 kDa laminin receptor (LRP/LR) acting as the cell surface receptor for the cellular Prion protein (PrPc) (1) and the infectious Prion protein (PrPsc) (2). We prooved that polyclonal antibodies were able to abolish PrPsc propagation in scrapie infected neuroblastoma cells (3), demonstrating that the disruption of the LRP-PrP interaction is a relevant strategy in therapies against TSEs. We injected the anti-LRP antibody W3 intraperitoneally into scrapie infected mice. Spleen analysis with respect to the PrPsc content will reveal whether this antibody is able to reduce PrPsc propagation. We did not observe any side effects after W3 application and monitor the survival times of the mice. Since a polyclonal antibody format is inappropriate for passive immunotransfer into humans, we developed single chain antibodies directed against the LRP/LR employing a phage display technique. Two scFvs termed N3 and S18 have been selected and characterized by epitope mapping, western blotting and FACS analysis. Both scFvs were able to recognize specifically the denatured form of LRP as well as the native form on the cell surface of BHK cells overexpressing LRP. The ability of these antibodies to interfere with the LRP-PrP interaction was proven by pull-down assays (4). In addition, a therapeutic effect of the scFvs on scrapie infected mice was investigated by passive immunotransfer. Although a significant reduction of peripheral PrPsc propagation in spleen of scrapie infected mice was observed, incubation times were not significantly prolonged. To circumvent the problem of fast renal antibody clearance, we are developing liposomes encapsulating the scFv and antibody secreting myotubes as alternative delivery systems.
(1) Gauczynski et al. (2001) EMBO J. 20, 5863-5875 (2) Gauczynski et al. (2006) J. infect. Diseases, in press (3) Leucht et al. (2003) EMBO rep 4, 290-295. (4) Rey et al., submitted.
AD C. Zuber, C. Rey, S. Weiss: Genzentrum - Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 Munich, Germany; S. Knackmuss, U. Reusch, M. Little: Affimed Therapeutics AG-Technologiepark- Im Neuheimer Feld 582, 69120 Heidelberg, Germany; C. Pace, G. Mitteregger: Zentrum für Neuropathologie und Prionforschung der LMU München, Feodor-Lynen-Str. 23, 81377 München, Germany; E-mail: weiss@lmb.uni-muenchen.de
SP englisch
PO Italien