Measuring prions causing bovine spongiform encephalopathy or chronic wasting disease by immunoassays and transgenic mice (original) (raw)

Nature Biotechnology volume 20, pages 1147–1150 (2002)Cite this article

Abstract

There is increasing concern over the extent to which bovine spongiform encephalopathy (BSE) prions have been transmitted to humans, as a result of the rising number of variant Creutzfeldt–Jakob disease (vCJD) cases. Toward preventing new transmissions, diagnostic tests for prions in livestock have been developed using the conformation-dependent immunoassay (CDI), which simultaneously measures specific antibody binding to denatured and native forms of the prion protein (PrP). We employed high-affinity recombinant antibody fragments (recFab) reacting with residues 95–105 of bovine (Bo) PrP for detection and another recFab that recognizes residues 132–156 for capture in the CDI. We report that the CDI is capable of measuring the disease-causing PrP isoform (PrPSc) in bovine brainstems with a sensitivity similar to that of end-point titrations in transgenic (Tg) mice expressing BoPrP. Prion titers were ∼107 ID50 units per gram of bovine brainstem when measured in Tg(BoPrP) mice, a figure ∼10 times greater than that determined by bioassay in cattle and ∼10,000× greater than in wild-type mice. We also report substantial differences in BoPrPSc levels in different areas of the obex region, where neuropathology has been consistently observed in cattle with BSE. The CDI was able to discriminate between PrPSc from BSE-infected cattle and Tg(BoPrP) mice as well as from chronic wasting disease (CWD)-infected deer and elk. Our findings argue that applying the CDI to livestock should considerably reduce human exposure to animal prions.

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References

  1. Mehlhorn, I. et al. High-level expression and characterization of a purified 142-residue polypeptide of the prion protein. Biochemistry 35, 5528–5537 (1996).
    Article CAS Google Scholar
  2. Safar, J. et al. Eight prion strains have PrPSc molecules with different conformations. Nat. Med. 4, 1157–1165 (1998).
    Article CAS Google Scholar
  3. Bruce, M. et al. Transmission of bovine spongiform encephalopathy and scrapie to mice: strain variation and the species barrier. Phil. Trans. R. Soc. Lond. B 343, 405–411 (1994).
    Article CAS Google Scholar
  4. Fraser, H., Bruce, M.E., Chree, A., McConnell, I. & Wells, G.A.H. Transmission of bovine spongiform encephalopathy and scrapie to mice. J. Gen. Virol. 73, 1891–1897 (1992).
    Article Google Scholar
  5. Moynagh, J., Schimmel, H. & Kramer, G.N. The evaluation of tests for the diagnosis of transmissible spongiform encephalopathy in bovines. 1–36 (European Commission, 1999).
    Google Scholar
  6. Deslys, J.P. et al. Screening slaughtered cattle for BSE. Nature 409, 476–478 (2001).
    Article CAS Google Scholar
  7. Wells, G.A.H. et al. Preliminary observations on the pathogenesis of experimental bovine spongiform encephalopathy (BSE): an update. Vet. Rec. 142, 103–106 (1998).
    Article CAS Google Scholar
  8. Scott, M.R. et al. Compelling transgenetic evidence for transmission of bovine spongiform encephalopathy prions to humans. Proc. Natl. Acad. Sci. USA 96, 15137–15142 (1999).
    Article CAS Google Scholar
  9. Peretz, D. et al. Strain-specified relative conformational stability of the scrapie prion protein. Protein Sci. 10, 854–863 (2001).
    Article CAS Google Scholar
  10. Schaller, O. et al. Validation of a western immunoblotting procedure for bovine PrPSc detection and its use as a rapid surveillance method for the diagnosis of bovine spongiform encephalopathy (BSE). Acta Neuropathol. 98, 437–443 (1999).
    Article CAS Google Scholar
  11. Biffiger, K. et al. Validation of a luminescence immunoassay for the detection of PrPSc in brain homogenate. J. Virol. Methods 101, 79–84 (2002).
    Article CAS Google Scholar
  12. Bosque, P.J. et al. Prions in skeletal muscle. Proc. Natl. Acad. Sci. USA 99, 3812–3817 (2002).
    Article CAS Google Scholar
  13. Prusiner, S.B. & Safar, J.G. Method of concentrating prion proteins in blood samples. US 6,166,187 (2000).
  14. Williamson, R.A. et al. Circumventing tolerance to generate autologous monoclonal antibodies to the prion protein. Proc. Natl. Acad. Sci. USA 93, 7279–7282 (1996).
    Article CAS Google Scholar
  15. Peretz, D. et al. A conformational transition at the N terminus of the prion protein features in formation of the scrapie isoform. J. Mol. Biol. 273, 614–622 (1997).
    Article CAS Google Scholar
  16. Peretz, D. et al. Antibodies inhibit prion propagation and clear cell cultures of prion infectivity. Nature 412, 739–743 (2001).
    Article CAS Google Scholar
  17. Burton, D.R. & Barbas, C.F. Human antibodies from combinatorial libraries. Adv. Immunol. 57, 191–280 (1994).
    Article CAS Google Scholar
  18. Johnsson, B., Lofas, S. & Lindquist, G. Immobilization of proteins to a carboxymethyldextran-modified gold surface for biospecific interaction analysis in surface plasmon resonance sensors. Anal. Biochem. 198, 268–277 (1991).
    Article CAS Google Scholar
  19. Dougherty, R. in Techniques in Experimental Virology (ed. Harris, R.J.C.) 169–224 (Academic Press, New York, 1964).
    Google Scholar
  20. Prusiner, S.B. et al. Immunologic and molecular biological studies of prion proteins in bovine spongiform encephalopathy. J. Infect. Dis. 167, 602–613 (1993).
    Article CAS Google Scholar
  21. Wells, G.A.H. & Wilesmith, J.W. The neuropathology and epidemiology of bovine spongiform encephalopathy. Brain Pathol. 5, 91–103 (1995).
    Article CAS Google Scholar

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Acknowledgements

The authors thank Elizabeth Williams (Wyoming State Veterinary Laboratory, University of Wyoming, Laramie) for providing the CWD samples, Hank Baron and Fred Feldman for encouragement and stimulating discussions, and the British Ministry of Agriculture, Fisheries and Food (MAFF) and VLA (UK) for supplying BSE brainstems. This work was supported by grants from the National Institutes of Health (AG02132, AG010770, NS22786, and NS14069) and MAFF (ES1756), as well as by gifts from the G. Harold and Leila Y. Mathers Charitable Foundation and from Centeon.

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Author notes

  1. Jeff Monaghan
    Present address: InPro Biotechnology, 870 Dubuque Avenue, South San Francisco, CA, 94080

Authors and Affiliations

  1. Institute for Neurodegenerative Diseases, University of California, San Francisco, 94143-0518, CA
    Jiri G. Safar, Michael Scott, Jeff Monaghan, Camille Deering, Svetlana Didorenko, Julie Vergara, Haydn Ball, Giuseppe Legname, Hana Serban, Darlene Groth & Stanley B. Prusiner
  2. Department of Neurology, University of California, San Francisco, 94143-0518, CA
    Jiri G. Safar, Michael Scott, Giuseppe Legname & Stanley B. Prusiner
  3. Department of Biochemistry and Biophysics, University of California, San Francisco, 94143-0518, CA
    Stanley B. Prusiner
  4. Departments of Immunology and Molecular Biology, The Scripps Research Institute, La Jolla, 92037, CA
    Estelle Leclerc, Laura Solforosi, Dennis R. Burton & R. Anthony Williamson

Authors

  1. Jiri G. Safar
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  2. Michael Scott
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  3. Jeff Monaghan
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  4. Camille Deering
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  5. Svetlana Didorenko
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  6. Julie Vergara
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  7. Haydn Ball
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  8. Giuseppe Legname
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  9. Estelle Leclerc
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  10. Laura Solforosi
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  11. Hana Serban
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  12. Darlene Groth
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  13. Dennis R. Burton
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  14. Stanley B. Prusiner
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  15. R. Anthony Williamson
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Corresponding authors

Correspondence toDennis R. Burton or Stanley B. Prusiner.

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Competing interests

J.G.S., J.M., G.L, H.S., D.G., and S.B.P. hold stock shares in InPro Biotechnology, which holds an exclusive license from the University of Calfornia for the conformation-dependent immunoassay (CDI).

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Safar, J., Scott, M., Monaghan, J. et al. Measuring prions causing bovine spongiform encephalopathy or chronic wasting disease by immunoassays and transgenic mice.Nat Biotechnol 20, 1147–1150 (2002). https://doi.org/10.1038/nbt748

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