The pathogenesis of bovine spongiform encephalopathy (BSE) infections in cattle has been studied in recent years by using highly sensitive transgenic-mouse bioassays. It has been shown that in this species, the BSE agent amplifies almost exclusively in the central and peripheral nervous system. Even in animals that were killed in the clinical end stage of the disease, the lymphoreticular system was shown to be free of the infectious agent. No other animal species investigated to date exhibits such a restricted BSE-infectivity distribution pattern. However, there is growing evidence for a radial spread of infection from the central nervous system (CNS) into the periphery during the late stages of the disease. In this study, we challenged transgenic mice overexpressing the bovine prion protein with homogenates prepared from a wide variety of tissue samples collected from BSE-infected cattle. As prion infections involve the conversion of the cellular prion protein into its abnormally folded isoform (PrP), we applied various detection methods, such as the purification of scrapie-associated fibrils, immunohistochemistry, and the protein misfolding cyclic amplification technique. Despite negative results using these highly sensitive biochemical methods, we were, for the first time, able to detect BSE infectivity in the tongue and in the nasal mucosa of terminally diseased BSE field cases as well as experimentally challenged cattle by transgenic-mouse bioassay. This shows that BSE infectivity can be present in the peripheral tissues of terminally diseased cattle, including tissues used for human consumption.


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  1. Andreoletti, O., Berthon, P., Marc, D., Sarradin, P., Grosclaude, J., van Keulen, L. J. M., Schelcher, F., Elsen, J. M. & Lantier, F.(2000). Early accumulation of PrPSc in gut-associated lymphoid and nervous tissues of susceptible sheep from a Romanov flock with natural scrapie. J Gen Virol 81, 3115–3126. [Google Scholar]
  2. Andreoletti, O., Simon, S., Lacroux, C., Morel, N., Tabouret, G., Chabert, A., Lugan, S., Corbiere, F., Ferre, P. & other authors(2004). PrPSc accumulation in myocytes from sheep incubating natural scrapie. Nat Med 10, 591–593.[CrossRef] [Google Scholar]
  3. Anonymous(2004). World Organisation for Animal Health (OIE), Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (Mammals, Birds and Bees), vol. 2, pp. 642–653. Paris. : World Organisation for Animal Health (OIE). [Google Scholar]
  4. Anonymous(2010).National Creutzfeldt-Jakob Disease Surveillance Unit (NCJDSU), Western General Hospital Edinburgh. Edinburgh, UK. : Worldwide vCJD statistics. [Google Scholar]
  5. Barron, R. M., Campbell, S. L., King, D., Bellon, A., Chapman, K. E., Williamson, R. A. & Manson, J. C.(2007). High titres of transmissible spongiform encephalopathy infectivity associated with extremely low levels of PrPScin vivo. J Biol Chem 282, 35878–35886.[CrossRef] [Google Scholar]
  6. Beekes, M. & McBride, P.(2000). Early accumulation of pathological PrP in the enteric nervous system and gut-associated lymphoid tissue of hamsters orally infected with scrapie. Neurosci Lett 278, 181–184.[CrossRef] [Google Scholar]
  7. Bosque, P. J., Ryou, C., Telling, G., Peretz, D., Legname, G., DeArmond, S. J. & Prusiner, S. B.(2002). Prions in skeletal muscle. Proc Natl Acad Sci U S A 99, 3812–3817.[CrossRef] [Google Scholar]
  8. Buschmann, A. & Groschup, M. H.(2005). Highly bovine spongiform encephalopathy-sensitive transgenic mice confirm the essential restriction of infectivity to the nervous system in clinically diseased cattle. J Infect Dis 192, 934–942.[CrossRef] [Google Scholar]
  9. Buschmann, A., Pfaff, E., Reifenberg, K., Müller, H. M. & Groschup, M. H.(2000). Detection of cattle-derived BSE prions using transgenic mice overexpressing bovine PrPC. Arch Virol Suppl 16, 75–86. [Google Scholar]
  10. Buschmann, A., Gretzschel, A., Biacabe, A.-G., Schiebel, K., Corona, C., Hoffmann, C., Eiden, M., Baron, T., Casalone, C. & Groschup, M. H.(2006). Atypical BSE cases in Germany. Vet Microbiol 117, 103–116.[CrossRef] [Google Scholar]
  11. Casalone, C., Corona, C., Crescio, M. I., Martucci, F., Mazza, M., Ru, G., Bozzetta, E., Acutis, P. L. & Caramelli, M.(2005). Pathological prion protein in the tongues of sheep infected with naturally occurring scrapie. J Virol 79, 5847–5849.[CrossRef] [Google Scholar]
  12. Casteleyn, C., Breugelmanns, S., Muylle, S., Van den Broeck, W. & Simoens, P.(2007). Consumption of bovine tongue and thymus: a risk to public health? Vlaams Diergeneeskundig Tijdschrift 76, 130–137 (in Dutch ). [Google Scholar]
  13. Castilla, J., Saa, P. & Soto, C.(2005a). Detection of prions in blood. Nat Med 11, 982–985. [Google Scholar]
  14. Castilla, J., Saa, P., Hetz, C. & Soto, C.(2005b).In vitro generation of infectious scrapie prions. Cell 121, 195–206.[CrossRef] [Google Scholar]
  15. Castilla, J., Saa, P., Morales, R., Abid, K., Maundrell, K. & Soto, C.(2006). Protein misfolding cyclic amplification for diagnosis and prion propagation studies. Methods Enzymol 412, 3–21. [Google Scholar]
  16. Espinosa, J. C., Morales, M., Castilla, J., Rogers, M. & Torres, J. M.(2007). Progression of prion infectivity in asymptomatic cattle after oral bovine spongiform encephalopathy challenge. J Gen Virol 88, 1379–1383.[CrossRef] [Google Scholar]
  17. Hardt, M., Baron, T. & Groschup, M. H.(2000). A comparative study of immunohistochemical methods for detecting abnormal prion protein with monoclonal and polyclonal antibodies. J Comp Pathol 122, 43–53.[CrossRef] [Google Scholar]
  18. Heggebo, R., Press, C. M., Gunnes, G., Lie, K. I., Tranulis, M. A., Uvlund, M., Groschup, M. H. & Landsverk, T.(2000). Distribution of prion protein in the ileal Peyer's patch of scrapie-free lambs and lambs naturally and experimentally exposed to the scrapie agent. J Gen Virol 81, 2327–2337. [Google Scholar]
  19. Herzog, C., Rivière, J., Lescoutra-Etchegaray, N., Charbonnier, A., Leblanc, V., Salès, N., Deslys, J. P. & Lasmézas, C. I.(2005). PrPTSE distribution in a primate model of variant, sporadic, and iatrogenic Creutzfeldt–Jakob disease. J Virol 79, 14339–14345.[CrossRef] [Google Scholar]
  20. Hoffmann, C., Ziegler, U., Buschmann, A., Weber, A., Kupfer, L., Oelschlegel, A., Hammerschmidt, B. & Groschup, M. H.(2007). Prions spread via the autonomic nervous system from the gut to the central nervous system in cattle incubating bovine spongiform encephalopathy. J Gen Virol 88, 1048–1055.[CrossRef] [Google Scholar]
  21. Kovacs, G. G., Kalev, O., Gelpi, E., Haberler, C., Wanschitz, J., Strohschneider, M., Molnar, M. J., Laszlo, L. & Budka, H.(2004). The prion protein in human neuromuscular diseases. J Pathol 204, 241–247.[CrossRef] [Google Scholar]
  22. Lasmezas, C. I., Deslys, J. P., Robain, O., Jeagly, A., Beringue, V., Reyrin, J. M., Fournier, J. G., Hauw, J. J., Rossier, J. & Dormont, D.(1997). Transmission of the BSE agent to mice in the absence of detectable abnormal prion protein. Science 275, 402–405.[CrossRef] [Google Scholar]
  23. Lezmi, S., Ronzon, F., Bencsik, A., Bedin, A., Calavas, D., Richard, Y., Simon, S., Grassi, J. & Baron, T.(2006). PrPd accumulation in organs of ARQ/ARQ sheep experimentally infected with BSE by peripheral routes. Acta Biochim Pol 53, 399–405. [Google Scholar]
  24. Manuelidis, E. E.(1975). Transmission of Creutzfeldt–Jakob disease from man to the guinea pig. Science 190, 571–572.[CrossRef] [Google Scholar]
  25. Piccardo, P., Manson, J. C., King, D., Ghetti, B. & Barron, R. M.(2007). Accumulation of prion protein in the brain that is not associated with transmissible disease. Proc Natl Acad Sci U S A 104, 4712–4717.[CrossRef] [Google Scholar]
  26. Press, C. M., Heggebo, R. & Espenes, A.(2004). Involvement of gut-associated lymphoid tissue of ruminants in the spread of transmissible spongiform encephalopathies. Adv Drug Deliv Rev 56, 885–899.[CrossRef] [Google Scholar]
  27. Rigter, A., Langeveld, J. P. M., Timmers-Parohi, D., Jacobs, J. G., Moonen, P. L. J. M. & Bossers, A.(2007). Mapping of possible prion protein self-interaction domains using peptide arrays. BMC Biochem 8, 6.[CrossRef] [Google Scholar]
  28. Saa, P., Castilla, J. & Soto, C.(2005). Cyclic amplification of protein misfolding and aggregation. Methods Mol Biol 299, 53–65. [Google Scholar]
  29. Saa, P., Castilla, J. & Soto, C.(2006). Ultraefficient replication of infectious prion by automated protein misfolding cyclic amplification. J Biol Chem 281, 35245–35252.[CrossRef] [Google Scholar]
  30. Schummer, A., Nickel, R., Habermehl, K. H., Vollmerhaus, B. & Wilkens, H.(1987). Eingeweide. In Lehrbuch der Anatomie der Haustiere, vol. 2, pp. 26. Edited by Nickel, R., Schummer, A., Seiferle, E. & Parey, Paul. Berlin and Hamburg. : Parey im MVS. [Google Scholar]
  31. Sun, R., Liu, Y., Zhang, H. & Manuelidis, L.(2008). Quantitative recovery of scrapie agent with minimal protein from highly infectious cultures. Viral Immunol 21, 293–302.[CrossRef] [Google Scholar]
  32. Tateishi, J., Sato, M., Doi, H. & Ohta, M.(1980). Experimental transmission of human subacute spongiform encephalopathy to small rodents. Acta Neuropathol 51, 127–134.[CrossRef] [Google Scholar]
  33. Terry, L. A., Marsh, S., Ryder, S. J., Hawkins, S. A., Wells, G. A. & Spencer, Y. I.(2003). Detection of disease-specific PrP in the distal ileum of cattle exposed orally to the agent of bovine spongiform encephalopathy. Vet Rec 152, 387–392.[CrossRef] [Google Scholar]
  34. Thomzig, A., Schulz-Schaeffer, W., Kratzel, C., Mai, J. & Beekes, M.(2004). Preclinical deposition of pathological prion protein in muscles of hamsters orally exposed to scrapie. J Clin Invest 113, 1465–1472.[CrossRef] [Google Scholar]
  35. van Keulen, L. J. M., Schreuder, B. E. C., Meloen, R. H., Mooij-Harkes, G., Vromans, M. E. V. & Langeveld, J. P. M.(1996). Immunohistochemical detection of prion protein in lymphoid tissues of sheep with natural scrapie. J Clin Microbiol 34, 1228–1231. [Google Scholar]
  36. Wells, G. A., Scott, A. C., Johnson, C. T., Gunning, R. F., Hancock, R. D., Jeffrey, M., Dawson, M. & Bradley, R.(1987). A novel progressive spongiform encephalopathy in cattle. Vet Rec 121, 419–420.[CrossRef] [Google Scholar]
  37. Wells, G. A. H., Spiropoulos, J., Hawkins, S. A. C. & Ryder, S. J.(2005). Pathogenesis of experimental bovine spongiform encephalopathy: preclinical infectivity in tonsil and observations on the distribution of lingual tonsil in slaughtered cattle. Vet Rec 156, 401–407.[CrossRef] [Google Scholar]
  38. Wilesmith, J. W., Wells, G. A., Cranwell, M. P. & Ryan, J. B.(1988). Bovine spongiform encephalopathy: epidemiological studies. Vet Rec 123, 638–644. [Google Scholar]

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