Development of antibody fragments for immunotherapy of prion diseases

Vincenza Campana (1) , Lorena Zentilin (2) , Ilaria Mirabile (3) , Agata Kranjc (3) , Philippe Casanova (1) , Mauro Giacca (2) , Stanley Prusiner (4) , Giuseppe Legname (3) , Chiara Zurzolo (1)

1 Trafic membranaire et Pathogénèse
2 ICGEB - International Centre for Genetic Engineering and Biotechnology
3 SISSA / ISAS - Scuola Internazionale Superiore di Studi Avanzati / International School for Advanced Studies
4 Institute for Neurodegenerative Diseases

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.

Mots clés: Life Sciences

Domaines: Microbiologie et Parasitologie

Vincenza Campana, Lorena Zentilin, Ilaria Mirabile, Agata Kranjc, Philippe Casanova, et al.. Development of antibody fragments for immunotherapy of prion diseases. Biochemical Journal, 2009, 418 (3), pp.507-515. ⟨10.1042/BJ20081541⟩. ⟨hal-00479077⟩