Molecular Medicine

, Volume 10, Issue 7–12, pp 104–111 | Cite as

Disease-Associated Prion Protein Elicits Immunoglobulin M Responses In Vivo

  • Mourad Tayebi
  • Perry Enever
  • Zahid Sattar
  • John Collinge
  • Simon Hawke


Prion diseases such as Creutzfeldt-Jakob disease are believed to result from the misfolding of a widely expressed normal cellular prion protein, PrPc. The resulting disease-associated isoforms, PrPSc, have much higher β-sheet content, are insoluble in detergents, and acquire relative resistance to proteases. Although known to be highly aggregated and to form amyloid fibrils, the molecular architecture of PrPSc is poorly understood. To date, it has been impossible to elicit antibodies to native PrPSc that are capable of recognizing PrPSc without denaturation, even in Prn-Po/o mice that are intolerant of it. Here we demonstrate that antibodies for native PrPc and PrPSc can be produced by immunization of Prn-Po/o mice with partially purified PrPc and PrPSc adsorbed to immunomagnetic particles using high-affinity anti-PrP monoclonal antibodies (mAbs). Interestingly, the polyclonal response to PrPSc was predominantly of the immunoglobulin M (IgM) isotype, unlike the immunoglobulin G (IgG) responses elicited by PrPc or by recombinant PrP adsorbed or not to immunomagnetic particles, presumably reflecting the polymeric structure of disease-associated prion protein. Although heat-denatured PrPSc elicited more diverse antibodies with the revelation of C-terminal epitopes, remarkably, these were also predominantly IgM suggesting that the increasing immunogenicity, acquisition of protease sensitivity, and reduction in infectivity induced by heat are not associated with dissociation of the PrP molecules in the diseased-associated protein. Adsorbing native proteins to immunomagnetic particles may have general applicability for raising polyclonal or monoclonal antibodies to any native protein, without attempting laborious purification steps that might affect protein conformation.



We thank Ray Young for preparation of figures and the animal team at the Charing Cross prion facility. This work is supported by grants from the Medical Research Council (UK).


  1. 1.
    Prusiner SB. (1994) Molecular biology and genetics of prion diseases. Philos. Trans. R. Soc. Lond. B Biol. Sci. 343:447–63.CrossRefGoogle Scholar
  2. 2.
    Gray F et al. (1994) Creutzfeldt-Jakob disease and cerebral amyloid angiopathy. Acta Neuropathol. (Berl) 88:106–11.CrossRefGoogle Scholar
  3. 3.
    Williams ES, Young S. (1980) Chronic wasting disease of captive mule deer: a spongiform encephalopathy. J. Wildl. Dis. 16:89–98.CrossRefGoogle Scholar
  4. 4.
    Wells GAH et al. (1987) A novel progressive spongiform encephalopathy in cattle. Vet. Record Oct. 31:419–20.CrossRefGoogle Scholar
  5. 5.
    Wilesmith JW, Wells GA, Cranwell MP, Ryan JB. (1988) Bovine spongiform encephalopathy: epidemiological studies. Vet. Record 123:638–44.Google Scholar
  6. 6.
    Simmons MM et al. (2000) Scrapie surveillance in Great Britain: results of an abattoir survey, 1997/98. Vet. Record 146:391–5.CrossRefGoogle Scholar
  7. 7.
    Collinge J, Sidle KCL, Meads J, Ironside J, Hill AF. (1996) Molecular analysis of prion strain variation and the etiology of ‘new variant’ CJD. Nature 383:685–90.CrossRefGoogle Scholar
  8. 8.
    Hill AF et al. (1997) The same prion strain causes vCJD and BSE. Nature 389:448–50.CrossRefGoogle Scholar
  9. 9.
    Bruce ME et al. (1997) Transmissions to mice indicate that ‘new variant’ CJD is caused by the BSE agent. Nature 389:498–501.CrossRefGoogle Scholar
  10. 10.
    Prusiner SB. (1998) The prion diseases. Brain Pathol. 8:499–513.CrossRefGoogle Scholar
  11. 11.
    Gajdusek DC. (1985) Hypothesis: interference with axonal transport of neurofilament as a common pathogenetic mechanism in certain diseases of the central nervous system. N. Engl. J. Med. 312:714–9.CrossRefGoogle Scholar
  12. 12.
    Prusiner SB et al. (1993) Immunologic and molecular biologic studies of prion proteins in bovine spongiform encephalopathy. J. Infect. Dis. 167:602–13.CrossRefGoogle Scholar
  13. 13.
    Bueler H et al. (1993) Mice devoid of PrP are resistant to scrapie. Cell 73:1339–47.CrossRefGoogle Scholar
  14. 14.
    Weissmann C et al. (1998) The use of transgenic mice in the investigation of transmissible spongiform encephalopathies. Rev. Sci. Tech. 17:278–90.CrossRefGoogle Scholar
  15. 15.
    Mabbott NA, Mackay F, Minns F, Bruce ME. (2000) Temporary inactivation of follicular dendritic cells delays neuroinvasion of scrapie. Nat. Med. 6:719–20.CrossRefGoogle Scholar
  16. 16.
    Hill AF et al. (1999) Investigation of variant Creutzfeldt-Jakob disease and other human prion diseases with tonsil biopsy samples. Lancet 353:183–9.CrossRefGoogle Scholar
  17. 17.
    O’Rourke KI et al. (2000) Preclinical diagnosis of scrapie by immunohistochemistry of third eyelid lymphoid tissue. J. Clin. Microbiol. 38:3254–9.PubMedPubMedCentralGoogle Scholar
  18. 18.
    Schreuder BE, Van Keulen LJ, Vromans ME, Langeveld JP, Smits MA. (1998) Tonsillar biopsy and PrPSc detection in the preclinical diagnosis of scrapie. Vet. Record 142:564–8.CrossRefGoogle Scholar
  19. 19.
    Hill AF, Zeidler M, Ironside J, Collinge J. (1997) Diagnosis of new variant Creutzfeldt-Jakob disease by tonsil biopsy. Lancet 349:99–100.CrossRefGoogle Scholar
  20. 20.
    Prusiner SB et al. (1993) Ablation of the prion protein (PrP) gene in mice prevents scrapie and facilitates production of anti-PrP antibodies. Proc. Natl. Acad. Sci. U.S.A. 90:10608–12.CrossRefGoogle Scholar
  21. 21.
    Prusiner SB et al. (1990) Transgenetic studies implicate interactions between homologous PrP isoforms in scrapie prion replication. Cell 63:673–86.CrossRefGoogle Scholar
  22. 22.
    Krasemann S, Groschup M, Hunsmann G, Bodemer W. (1996) Induction of antibodies against human prion proteins (PrP) by DNA-mediated immunization of PrP mice. J. Immunol. Methods 199:109–18.CrossRefGoogle Scholar
  23. 23.
    Korth C et al. (1997) Prion (PrPSc)-specific epitope defined by a monoclonal antibody. Nature 390:74–7.CrossRefGoogle Scholar
  24. 24.
    Paramithiotis E et al. (2003) A prion protein epitope selective for the pathologically misfolded conformation. Nat. Med. 9:893–9.CrossRefGoogle Scholar
  25. 25.
    Maissen M, Roeckl C, Glatzel M, Goldmann W, Aguzzi A. (2001) Plasminogen binds to disease-associated prion protein of multiple species. Lancet 357:2026–8.CrossRefGoogle Scholar
  26. 26.
    Weiss S et al. (1997) RNA aptamers specifically interact with the prion protein PrP. J. Virol. 71:8790–7.PubMedPubMedCentralGoogle Scholar
  27. 27.
    Soto C et al. (2000) Reversion of prion protein conformational changes by synthetic β-sheet breaker peptides. Lancet 355:192–7.CrossRefGoogle Scholar
  28. 28.
    Safar J et al. (1998) Eight prion strains PrPSc molecules with different conformations. Nat. Med. 4:1157–65.CrossRefGoogle Scholar
  29. 29.
    Porter DD, Porter HG, Cox NA. (1973) Failure to demonstrate a humoral immune response to scrapie infection in mice. J. Immunol. 111:1407–10.PubMedGoogle Scholar
  30. 30.
    Aguzzi A. (1998) Protein conformation dictates prion strain. Nat. Med. 4:1125–6.CrossRefGoogle Scholar
  31. 31.
    McBride PA, Eikelenboom P, Kraal G, Fraser H, Bruce ME. (1992) PrP protein is associated with follicular dendritic cells of spleens and lymph nodes in uninfected and scrapie-infected mice. J. Pathol. 168:413–8.CrossRefGoogle Scholar
  32. 32.
    Peretz D et al. (1997) A conformational transition at the N terminus of the prion protein features in formation of the scrapie isoform. J. Mol. Biol. 273:614–22.CrossRefGoogle Scholar
  33. 33.
    Kascsak RJ et al. (1987) Mouse polyclonal and monoclonal antibody to scrapieassociated fibril proteins. J. Virol. 61:3688–93.PubMedPubMedCentralGoogle Scholar
  34. 34.
    Beringue V et al. (2003) Regional heterogeneity of cellular prion protein isoforms in the mouse brain. Brain 126:2065–73.CrossRefGoogle Scholar
  35. 35.
    Mond JJ, Vos Q, Lees A, Snapper CM. (1995) T cell independent antigens. Curr. Opin. Immunol. 7:349–54.CrossRefGoogle Scholar
  36. 36.
    Bueler H et al. (1992) Normal development and behavior of mice lacking the neuronal cell-surface PrP protein. Nature 356:577–582.CrossRefGoogle Scholar
  37. 37.
    Brandner S et al. (1996) Normal host prion protein (PrPC) is required for scrapie spread within the central nervous system. Proc. Natl. Acad. Sci. U.S.A. 93: 13148–51.CrossRefGoogle Scholar
  38. 38.
    Jackson GS et al. (1999) Reversible conversion of monomeric human prion protein between native and fibrilogenic conformations. Science 283:1935–7.CrossRefGoogle Scholar
  39. 39.
    Laemmli UK. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–5.CrossRefGoogle Scholar
  40. 40.
    Anstee DJ et al. (1991) New monoclonal antibodies in CD44 and CD58: their use to quantify CD44 and CD58 on normal human erythrocytes and to compare the distribution of CD44 and CD58 in human tissues. Immunology 74: 197–205.PubMedPubMedCentralGoogle Scholar
  41. 41.
    Taylor DM, Fernie K, Steele PJ, McConnell I, Somerville RA. (2002) Thermostability of mouse-passaged BSE and scrapie is independent of host PrP genotype: implications for the nature of the causal agents. J. Gen. Virol. 83:3199–204.CrossRefGoogle Scholar
  42. 42.
    Serban D, Taraboulos A, DeArmond SJ, Prusiner SB. (1990) Rapid detection of Creutzfeldt-Jakob disease and scrapie prion proteins. Neurology 40:110–7.CrossRefGoogle Scholar
  43. 43.
    Krasemann S, Jürgens T, Bodemer W. (1999) Generation of monoclonal antibodies against prion proteins with an unconventional nucleic acid-based immunization strategy. J. Biotechnol. 73:119–29.CrossRefGoogle Scholar
  44. 44.
    Jackson GS et al. (1999) Multiple folding pathways for heterologously expressed human prion protein. Biochim. Biophys. Acta 1431:1–13.CrossRefGoogle Scholar
  45. 45.
    Hawke S, Willcox N, Harcourt G, Vincent A, Newsom-Davis J. (1992) Stimulation of human T cells by sparse antigens captured on immunomagnetic particles. J. Immunol. Methods 155:41–8.CrossRefGoogle Scholar
  46. 46.
    Hawke S et al. (1996) Autoimmune T cells in myasthenia gravis: heterogeneity and potential for specific immunotargeting. Immunol. Today 17:307–11.CrossRefGoogle Scholar
  47. 47.
    Harmeyer S, Pfaff E, Groschup MH. (1998) Synthetic peptide vaccines yield monoclonal antibodies to cellular and pathological prion proteins of ruminants. J. Gen. Virol. 79:937–45.CrossRefGoogle Scholar
  48. 48.
    Mond JJ, Vos Q, Lees A, Snapper CM. (1995) T cell independent antigens. Curr. Opin. Immunol. 7:349–54.CrossRefGoogle Scholar
  49. 49.
    O’Nuallain B, Wetzel R. (2002) Conformational Abs recognizing a generic amyloid fibril epitope. Proc. Natl. Acad. Sci. U.S.A. 99:1485–90.CrossRefGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2004

Authors and Affiliations

  • Mourad Tayebi
    • 1
    • 2
  • Perry Enever
    • 1
  • Zahid Sattar
    • 1
  • John Collinge
    • 1
    • 2
  • Simon Hawke
    • 1
    • 2
  1. 1.MRC Prion Unit and Department of Neurodegenerative Disease, Institute of NeurologyUniversity College LondonLondonUK
  2. 2.Department of Neurogenetics, Division of Neuroscience and Psychological Medicine, Faculty of MedicineImperial CollegeLondonUK

Personalised recommendations