Clearance and prevention of prion infection in cell culture by anti-PrP antibodies.
Prion diseases are transmissible and invariably fatal neurodegenerative disorders associated with a conformational transformation of the cellular prion protein (PrP(C)) into a self-replicating and proteinase K (PK)-resistant conformer, scrapie PrP (PrP(Sc)). Humoral immunity may significantly prolong the incubation period and even prevent disease in murine models of prionoses. However, the mechanism(s) of action of anti-PrP monoclonal antibodies (Mabs) remain(s) obscure. The murine neuroblastoma N2a cell line, infected with the 22L mouse-adapted scrapie strain, was used to screen a large library of Mabs with similar binding affinities to PrP, to identify those antibodies which could clear established infection and/or prevent infection de novo. Three Mabs were found capable of complete and persistent clearing of already-infected N2a cells of PrP(Sc). These antibodies were 6D11 (generated to PK-resistant PrP(Sc) and detecting PrP residues 93-109), and 7H6 and 7A12, which were raised against recombinant PrP and react with neighbouring epitopes of PrP residues 130-140 and 143-155, respectively. Mabs were found to interact with PrP(Sc) formation both on the cell surface and after internalization in the cytosol. Treatment with Mabs was not associated with toxicity nor did it result in decreased expression of PrP(C). Both preincubation of N2a cells with Mabs prior to exposure to 22L inoculum and preincubation of the inoculum with Mabs prior to infecting N2a cells resulted in a significant reduction in PrP(Sc) levels. Information provided in these studies is important for the rational design of humoral immune therapy for prion infection in animals and eventually in humans.
Pankiewicz
Joanna
J
Department of Neurology, New York University School of Medicine, 550 First Avenue, New York, NY 10016, USA.
Prelli
Frances
F
Sy
Man-Sun
MS
Kascsak
Richard J
RJ
Kascsak
Regina B
RB
Spinner
Daryl S
DS
Carp
Richard I
RI
Meeker
Harry C
HC
Sadowski
Marcin
M
Wisniewski
Thomas
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000915:Antibody Affinity; D002478:Cells, Cultured; D018604:Epitope Mapping; D016056:Immunodominant Epitopes; D051379:Mice; D018621:PrPC Proteins / Q000187:drug effects* / Q000276:immunology; D018620:PrPSc Proteins / Q000187:drug effects* / Q000276:immunology / Q000472:pathogenicity; D017096:Prion Diseases / Q000276:immunology* / Q000517:prevention & control*
D000911:Antibodies, Monoclonal; D016056:Immunodominant Epitopes; D018621:PrPC Proteins; D018620:PrPSc Proteins