Paracrine inhibition of prion propagation by anti-PrP single-chain Fv miniantibodies.
Prion diseases are characterized by the deposition of PrP(Sc), an abnormal form of the cellular prion protein PrP(C). A growing body of evidence suggests that antibodies to PrP(C) can antagonize deposition of PrP(Sc). However, host tolerance hampers the induction of immune responses to PrP(C), and cross-linking of PrP(C) by bivalent anti-PrP antibodies is neurotoxic. In order to obviate these problems, we explored the antiprion potential of recombinant single-chain antibody (scFv) fragments. scFv fragments derived from monoclonal anti-PrP antibody 6H4, flagged with c-myc and His6 tags, were correctly processed and secreted by mammalian RD-4 rhabdomyosarcoma cells. When cocultured with cells secreting anti-PrP scFv, chronically prion-infected neuroblastoma cells ceased to produce PrP(Sc), even if antibody-producing cells were physically separated from target cells in transwell cultures. Expression of scFv with irrelevant specificity, or of similarly tagged molecules, was not curative. Therefore, eukaryotically expressed scFv exerts a paracrine antiprion activity. The effector functions encoded by immunoglobulin constant domains are unnecessary for this effect. Because of their small size and their monovalent binding, scFv fragments may represent candidates for gene transfer-based immunotherapy of prion diseases.
Donofrio
Gaetano
G
Institute of Neuropathology, University Hospital Zurich, Schmelzbergstrasse 12, CH-8091 Zürich, Switzerland.
Heppner
Frank L
FL
Polymenidou
Magdalini
M
Musahl
Christine
C
Aguzzi
Adriano
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D017931:DNA Primers; D006801:Humans; D007128:Immunoglobulin Fragments / Q000494:pharmacology*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011328:Prions / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D012208:Rhabdomyosarcoma
D017931:DNA Primers; D007128:Immunoglobulin Fragments; D011328:Prions; C055966:immunoglobulin Fv