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Antibodies inhibit prion propagation and clear cell cultures of prion infectivity

Nature volume 412, pages 739–743 (2001)Cite this article

Abstract

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 prions1,2. 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.

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Figure 1: Dose-dependent inhibition of PrPSc formation in cells by prion protein-specific recombinant antibody Fabs.
Figure 2: Elimination of PrPSc from prion-infected cells treated with antibodies.
Figure 3: Time course of antibody-mediated PrPSc clearance.
Figure 4: Regions of sequence recognized by Fabs D13 (blue), D18 (red), and R1 and R2 (green) superimposed onto two views of the structure of recombinant prion protein (90–231) determined by nuclear magnetic resonance27.

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Acknowledgements

We thank J. Safar, S. DeArmond, F. Cohen and P. Dazin for discussions and K. Crossin for anti-N-CAM antibody. This work was supported by grants from the National Institutes of Health to R.A.W., D.R.B. and S.B.P.

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Author notes

  1. Kiotoshi Kaneko

    Present address: National Institute of Neuroscience, Tokyo, 187-8502, Japan

Authors and Affiliations

  1. Institute for Neurodegenerative Diseases, University of California,, San Francisco, 94143-0518, California, USA

    David Peretz, Kiotoshi Kaneko, Julie Vergara, Gerold Schmitt-Ulms, Ingrid R. Mehlhorn, Giuseppe Legname & Stanley B. Prusiner

  2. Department of Neurology, University of California, San Francisco, 94143-0518, California, USA

    Stanley B. Prusiner

  3. Department of Biochemistry and Biophysics, University of California, San Francisco, 94143-0518, California, USA

    Stanley B. Prusiner

  4. Department of Immunology, The Scripps Research Institute, La Jolla, 92037, California, USA

    R. Anthony Williamson, Estelle Leclerc & Dennis R. Burton

  5. Department of Molecular Biology, The Scripps Research Institute, La Jolla, 92037, California, USA

    Dennis R. Burton

  6. Department of Biochemistry, The Glycobiology Institute, University of Oxford, Oxford, OX1 3QU, UK

    Mark R. Wormald, Pauline M. Rudd & Raymond A. Dwek

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Corresponding author

Correspondence to R. Anthony Williamson.

Supplementary information

Table 1. Epitopes recognized and dissociation constants for the binding of PrP-specific Fabs to recombinant PrP(29-231) refolded into an a-helical conformation. Binding constants were determined by surface plasmon resonance.
Full size table

Figure 5

(JPG 26.2 KB)

Levels of PrPC in antibody-treated ScN2a cells determined by immunoblot. ScN2a cultures were treated for a period of 7 days with 10 mg/ml of Fabs D13, D18, R72, R1, R2, E123 or E149, or passaged in the absence of antibody.

Figure 6

(JPG 35.4 KB)

PrP-specific antibody Fab binding to the surface of ScN2a cells. The reactivity of PrP-specific Fabs D13, D18, R72, R1 and R2 with the surface of ScN2a cells was evaluated using flow cytometry. The data represent the mean of three experiments.

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Peretz, D., Williamson, R., Kaneko, K. et al. Antibodies inhibit prion propagation and clear cell cultures of prion infectivity. Nature 412, 739–743 (2001). https://doi.org/10.1038/35089090

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