Antibodies inhibit prion propagation and clear cell cultures of prion infectivity

Prions are the transmissible pathogenic agents responsible for diseases such as scrapie and bovine spongiform encephalopathy. In the favoured model of prion replication, direct interaction between the pathogenic prion protein (PrPSc) template and endogenous cellular prion protein (PrPC) is proposed to drive the formation of nascent infectious prions. Reagents specifically binding either prion-protein conformer may interrupt prion production by inhibiting this interaction. We examined the ability of several recombinant antibody antigen-binding fragments (Fabs) to inhibit prion propagation in cultured mouse neuroblastoma cells (ScN2a) infected with PrPSc. Here we show that antibodies binding cell-surface PrPC inhibit PrPSc formation in a dose-dependent manner. In cells treated with the most potent antibody, Fab D18, prion replication is abolished and pre-existing PrPSc is rapidly cleared, suggesting that this antibody may cure established infection. The potent activity of Fab D18 is associated with its ability to better recognize the total population of PrPC molecules on the cell surface, and with the location of its epitope on PrPC. Our observations support the use of antibodies in the prevention and treatment of prion diseases and identify a region of PrPC for drug targeting.

[1]  F. Cohen,et al.  Dominant-Negative Inhibition of Prion Formation Diminished by Deletion Mutagenesis of the Prion Protein , 2000, Journal of Virology.

[2]  D R Burton,et al.  Circumventing tolerance to generate autologous monoclonal antibodies to the prion protein. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[3]  S. Priola,et al.  Efficient Conversion of Normal Prion Protein (PrP) by Abnormal Hamster PrP Is Determined by Homology at Amino Acid Residue 155 , 2001, Journal of Virology.

[4]  B. Caughey,et al.  Inhibition of protease-resistant prion protein formation by porphyrins and phthalocyanines. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[5]  F E Cohen,et al.  Solution structure of a 142-residue recombinant prion protein corresponding to the infectious fragment of the scrapie isoform. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[6]  F. Cohen,et al.  Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Stanley B. Prusiner,et al.  Nobel Lecture: Prions , 1998 .

[8]  F E Cohen,et al.  A conformational transition at the N terminus of the prion protein features in formation of the scrapie isoform. , 1997, Journal of molecular biology.

[9]  P. Parren,et al.  In vitro characterization of five humanized OKT3 effector function variant antibodies. , 2000, Cellular immunology.

[10]  B. Chesebro,et al.  Characterization of scrapie infection in mouse neuroblastoma cells. , 1987, The Journal of general virology.

[11]  G. J. Raymond,et al.  Sulfated polyanion inhibition of scrapie-associated PrP accumulation in cultured cells , 1993, Journal of virology.

[12]  B. Chesebro,et al.  A single hamster PrP amino acid blocks conversion to protease-resistant PrP in scrapie-infected mouse neuroblastoma cells , 1995, Journal of virology.

[13]  F. Cohen,et al.  Prion Protein of 106 Residues Creates an Artificial Transmission Barrier for Prion Replication in Transgenic Mice , 1999, Cell.

[14]  B. Chesebro,et al.  Species-Independent Inhibition of Abnormal Prion Protein (PrP) Formation by a Peptide Containing a Conserved PrP Sequence , 1999, Journal of Virology.

[15]  S. Prusiner,et al.  Propagation of prions with artificial properties in transgenic mice expressing chimeric PrP genes , 1993, Cell.

[16]  F. Cohen,et al.  Elimination of prions by branched polyamines and implications for therapeutics. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  S. Prusiner,et al.  Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells , 1990, The Journal of cell biology.

[18]  Leonard G. Presta,et al.  Mapping of the C1q Binding Site on Rituxan, a Chimeric Antibody with a Human IgG1 Fc , 2000, The Journal of Immunology.

[19]  F. Cohen,et al.  Mimicking dominant negative inhibition of prion replication through structure-based drug design. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[20]  G. Edelman,et al.  Neural cell adhesion molecules in rodent brains isolated by monoclonal antibodies with cross-species reactivity. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[21]  S. Prusiner,et al.  Scrapie-infected murine neuroblastoma cells produce protease-resistant prion proteins , 1988, Journal of virology.

[22]  S. Prusiner,et al.  Immobilized prion protein undergoes spontaneous rearrangement to a conformation having features in common with the infectious form , 2001, The EMBO journal.

[23]  S. Prusiner,et al.  Evidence for the Conformation of the Pathologic Isoform of the Prion Protein Enciphering and Propagating Prion Diversity , 1996, Science.

[24]  R. Dwek,et al.  A statistical analysis of N- and O-glycan linkage conformations from crystallographic data. , 1999, Glycobiology.

[25]  S. Prusiner,et al.  Prion glycoprotein: structure, dynamics, and roles for the sugars. , 2001, Biochemistry.

[26]  R. Motter,et al.  Immunization with amyloid-β attenuates Alzheimer-disease-like pathology in the PDAPP mouse , 1999, Nature.

[27]  P. Lansbury,et al.  Species specificity in the cell-free conversion of prion protein to protease-resistant forms: a model for the scrapie species barrier. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Clemencia Pinilla,et al.  Mapping the Prion Protein Using Recombinant Antibodies , 1998, Journal of Virology.

[29]  R. Motter,et al.  Peripherally administered antibodies against amyloid β-peptide enter the central nervous system and reduce pathology in a mouse model of Alzheimer disease , 2000, Nature Medicine.