1. Development of antibody fragments for immunotherapy of prion diseases
- Author
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Chiara Zurzolo, Philippe Casanova, Ilaria Mirabile, Giuseppe Legname, Vincenza Campana, Lorena Zentilin, Mauro Giacca, Agata Kranjc, Stanley B. Prusiner, V., Campana, L., Zentilin, I., Mirabile, A., Kranjc, P., Casanova, Giacca, Mauro, S. B., Prusiner, G., Legname, C., Zurzolo, Trafic membranaire et Pathogénèse, Institut Pasteur [Paris], International Centre for Genetic Engineering and Biotechnology (ICGEB), Scuola Internazionale Superiore di Studi Avanzati / International School for Advanced Studies (SISSA / ISAS), Institute for Neurodegenerative Diseases, University of California [San Francisco] (UCSF), University of California-University of California, Institut Pasteur [Paris] (IP), University of California [San Francisco] (UC San Francisco), University of California (UC)-University of California (UC), Campana, V., Zentilin, L., Mirabile, I., Kranjc, A., Casanova, P., Giacca, M., Prusiner, S., Legname, G., and Zurzolo, Chiara
- Subjects
PrPSc Proteins ,animal diseases ,Adeno-associated Viru ,Immunoglobulin Variable Region ,Scrapie ,Active immunotherapy ,medicine.disease_cause ,Biochemistry ,Prion Diseases ,immunology ,Mice ,0302 clinical medicine ,genetics ,Adeno-associated virus ,Cells, Cultured ,0303 health sciences ,Cultured ,biology ,Life Sciences ,Dependovirus ,Animals, Antibodies ,immunology, Cells ,Cultured, Dependovirus ,genetics, Genetic Vectors, Immunoglobulin Variable Region ,immunology/therapeutic use, Immunotherapy ,methods, Lentivirus ,genetics, Mice, PrPC Proteins ,immunology, PrPSc Proteins ,immunology, Prion Diseases ,therapy, Prions, Scrapie ,therapy, Viral Fusion Proteins ,3. Good health ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,adeno-associated virus ,immunotherapy ,lentivirus ,cellular prion protein (PrPC) ,scrapie prion protein (PrPSc) ,single-chain variable fragment (scFv) ,immunology/therapeutic use ,Lentivirus ,Animals, Antibodie ,Immunotherapy ,Antibody ,therapy, Viral Fusion Protein ,methods, Lentiviru ,Prions ,Cells ,Genetic Vectors ,Lentiviru ,Antibodies ,Virus ,methods ,ScFv ,03 medical and health sciences ,immunology, Prion Disease ,Immune system ,medicine ,Animals ,PrPC Proteins ,Molecular Biology ,genetics, Mice, PrPC Protein ,030304 developmental biology ,therapy ,PrPC ,Cell Biology ,biology.organism_classification ,Virology ,Cultured, Dependoviru ,nervous system diseases ,immunology, Cell ,Cell culture ,biology.protein ,immunology, PrPSc Protein ,Viral Fusion Proteins ,030217 neurology & neurosurgery ,PrPSc - Abstract
International audience; Prions are infectious proteins responsible for a group of fatal neurodegenerative diseases called transmissible spongiform encephalopathies (TSEs), or prion diseases. In mammals, prions reproduce themselves by recruiting the normal cellular protein (PrPC) and inducing its conversion to the disease-causing isoform denominated PrPSc. Recently, anti-prion antibodies have been shown to permanently cure prion-infected cells. However, the inability of full-lenght antibodies and proteins to cross the blood-brain barrier (BBB) hampers their use in the therapy of TSEs in vivo. Alternatively, brain delivery of prion-specific scFv by adeno-associated virus (AAV) transfer delays the onset of the disease in infected mice, although protection is not complete. We investigated the anti-prion effects of a recombinant anti-PrP (D18) single-chain variable fragment (scFv) by direct addition to scrapie infected cell cultures or by infection with both lentivirus and adeno-associated virus (AAV) transducing vectors. We show that recombinant anti-PrP scFv is able to reduce proteinase K-resistant PrP content in infected cells. In addition, we demonstrate that lentiviruses are more efficient than AAV in gene transferring of the anti-PrP scFv gene and in reducing PrPSc content in infected neuronal cell lines. Finally, we have used a bioinformatic approach to construct a structural model of D18scFv-PrPC complex. Interestingly, according to the docking results, ArgPrP151 (Arg151 from prion protein) is the key residue for the interactions with D18scFv, anchoring the PrPC to the cavity of the antibody. Taken together, these results indicate that combined passive and active immunotherapy targeting PrP might be promising strategies for therapeutic intervention in prion diseases.
- Published
- 2009