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Measuring prions causing bovine spongiform encephalopathy or chronic wasting disease by immunoassays and transgenic mice

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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Figure 1: Similar sensitivity of the CDI and bioassays in Tg(BoPrP)Prnp0/0 mice for BSE prions.
Figure 2: Statistical evaluations of the direct and sandwich automated CDI.
Figure 3: Transverse distribution of PrPSc in bovine brainstems at the level of the obex as determined by CDI analysis correlates with known BSE pathology.
Figure 4: Different conformational characteristics of ungulate prion strains revealed by direct CDI.

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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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  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

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Corresponding authors

Correspondence to Dennis 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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