Copurification of Sp33–37 and Scrapie Agent from Hamster Brain Prior to Detectable Histopathology and Clinical Disease Free

Abstract

Studies were conducted to determine whether accumulation of the scrapie agent protein Sp33–37 in brain correlated with the appearance of the scrapie agent or with pathology. The concentrations of the scrapie agent and Sp33–37 were measured in purified fraction P isolated from hamster brains at weekly intervals after inoculation. The scrapie agent concentration in fraction P was approximately 10 LD/g brain 1 day post-inoculation and increased to 10LD/g at day 77. Sp33–37 was first detected in P at day 21, when the agent titre was 10 LD/g. Sp33–37 concentration increased in concert with the scrapie agent concentration, although the apparent rate of increase was somewhat lower for the protein than for the agent. The histopathological evidence of disease, consisting of mild vacuolation and gliosis, was first seen at 35 days, but was not conspicuous until 49 to 56 days post-inoculation. Vacuolation and gliosis increased until termination of the experiment at day 77. Amyloid plaques were first detected at 56 days and were widespread at day 77. Clinical disease was first seen in these animals at day 66, with an average onset at day 71. Control animals inoculated with buffer alone showed some mild gliosis, but were otherwise normal. The fact that Sp33–37 purified with the scrapie agent isolated from brain 14 days prior to detectable (light microscopic) pathology supports the theory that Sp33–37 is the major structural component of the scrapie agent and not solely a product of the pathology.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-12-2905
1991-12-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/12/JV0720122905.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-12-2905&mimeType=html&fmt=ahah

References

  1. Aiken J. M., Williamson J. L., Marsh R. F. 1989; Evidence of mitochondrial involvement in scrapie infection. Journal of Virology 63:1686–1694
    [Google Scholar]
  2. Barry R. A., Prusiner S. B. 1987; Immunology of prions. In Prions: Novel Infectious Pathogens Causing Scrapie and Creutzfeldt-Jakob Disease pp 239–275 Edited by Prusiner S. B., McKinley M. P. San Diego: Academic Press;
    [Google Scholar]
  3. Barry R. A., McKinley M. P., Bendheim P. E., Lewis G. K., DeArmond S. J., Prusiner S. B. 1985; Antibodies to the scrapie protein decorate prion rods. Journal of Immunology 135:603–613
    [Google Scholar]
  4. Bendheim P. E., Bolton D. C. 1986; A 54-kDa normal cellular protein may be the precursor of the scrapie agent protease-resistant protein. Proceedings of the National Academy of Sciences, U.S.A. 83:2214–2218
    [Google Scholar]
  5. Bendheim P. E., Barry R. A., DeArmond S. J., Stites D. P., Prusiner S. B. 1984; Antibodies to a scrapie prion protein. Nature, London 310:418–421
    [Google Scholar]
  6. Bendheim P. E., Potempska A., Kascsak R. J., Bolton D. C. 1988b; Purification and partial characterization of the normal cellular homologue of the scrapie agent protein. Journal of Infectious Diseases 158:1198–1208
    [Google Scholar]
  7. Blum H., Beier H., Gross H. J. 1987; Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99
    [Google Scholar]
  8. Bolton D. C., Bendheim P. E. 1988; A modified host protein model of scrapie. Ciba Foundation Symposium 135:164–181
    [Google Scholar]
  9. Bolton D. C., McKinley M. P., Prusiner S. B. 1982; Identification of a protein that purifies with the scrapie prion. Science 218:1309–1311
    [Google Scholar]
  10. Bolton D. C., Meyer R. K., Prusiner S. B. 1985; Scrapie PrP 27–30 is a sialoglycoprotein. Journal of Virology 53:596–606
    [Google Scholar]
  11. Bolton D. C., Bendheim P. E., Marmorstein A. D., Potempska A. 1987a; Isolation and structural studies of the intact scrapie agent protein. Archives of Biochemistry and Biophysics 258:579–590
    [Google Scholar]
  12. Bolton D. C., McKinley M. P., Prusiner S. B. 1987b; Properties and characteristics of scrapie PrP 27–30 protein. In Prions: Novel Infectious Pathogens Causing Scrapie and Creutzfeldt-Jakob Disease pp 173–196 Edited by Prusiner S. B., McKinley M. P. San Diego: Academic Press;
    [Google Scholar]
  13. Braig H. R., Diringer H. 1985; Scrapie: concept of a virus-induced amyloidosis of the brain. EMBO Journal 4:2309–2312
    [Google Scholar]
  14. Brown P., Liberski P. P., Wolff A., Gajdusek D. C. 1990; Conservation of infectivity in purified fibrillary extracts of scrapie-infected hamster brain after sequential enzymatic digestion or polyacrylamide gel electrophoresis. Proceedings of the National Academy of Sciences, U.S.A. 87:7240–7244
    [Google Scholar]
  15. Burnette W. N. 1981; ‘Western blotting’: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Analytical Biochemistry 112:195–203
    [Google Scholar]
  16. Carp R. I., Kascsak R. J., Wisniewski H. M., Merz P. A., Rubenstein R., Bendheim P. E., Bolton D. C. 1989; The nature of the unconventional slow infection agents remains a puzzle. Alzheimer Disease and Associated Disorders 3:79–99
    [Google Scholar]
  17. Czub M., Braig H. R., Diringer H. 1986; Pathogenesis of scrapie: study of the temporal development of clinical symptoms, of infectivity titres and scrapie-associated fibrils in brains of hamsters infected intraperitoneally. Journal of General Virology 67:2005–2009
    [Google Scholar]
  18. Czub M., Braig H. R., Diringer H. 1988; Replication of the scrapie agent in hamsters infected intracerebrally confirms the pathogenesis of an amyloid-inducing virosis. Journal of General Virology 69:1753–1756
    [Google Scholar]
  19. DeArmond S. J., McKinley M. P., Barry R. A., Braunfeld M. B., McColloch J. R., Prusiner S. B. 1985; Identification of prion amyloid filaments in scrapie-infected brain. Cell 41:221–235
    [Google Scholar]
  20. DeArmond S. J., Mobley W. C., DeMott D. L., Barry R. A., Beckstead J. H., Prusiner S. B. 1987; Changes in the localization of brain prion proteins during scrapie infection [published erratum appears in Neurology (1987) 37, 1770]. Neurology 37:1271–1280
    [Google Scholar]
  21. Dees C., German T. L., Wade W. F., Marsh R. F. 1985; Characterization of proteins in membrane vesicles from scrapie-infected hamster brain. Journal of General Virology 66:851–859
    [Google Scholar]
  22. Diedrich J. F., Bendheim P. E., Kim K. S., Carp R. I., Haase A. T. 1991; Scrapie-associated prion protein accumulates in astrocytes during scrapie infection. Proceedings of the National Academy of Sciences, U.S.A. 88:375–379
    [Google Scholar]
  23. Gabizon R., McKinley M. P., Prusiner S. B. 1987; Purified prion proteins and scrapie infectivity copartition into liposomes. Proceedings of the National Academy of Sciences, U.S.A. 84:4017–4021
    [Google Scholar]
  24. Gabizon R., McKinley M. P., Groth D., Prusiner S. B. 1988; Immunoaffinity purification and neutralization of scrapie prion infectivity. Proceedings of the National Academy of Sciences, U.S.A. 85:6617–6621
    [Google Scholar]
  25. Griffith J. S. 1967; Self-replication and scrapie. Nature, London 215:1043–1044
    [Google Scholar]
  26. Hope I., Morton L. J., Farquhar C. F., Multhaup G., Beyreuther K., Kimberlin R. H. 1986; The major polypeptide of scrapie-associated fibrils (SAF) has the same size, charge distribution and N-terminal protein sequence as predicted for the normal brain protein (PrP). EMBO Journal 5:2591–2597
    [Google Scholar]
  27. Hsiao K. K., Scott M., Foster D., Groth D. F., DeArmond S. J., Prusiner S. B. 1990; Spontaneous neurodegeneration in transgenic mice with mutant prion protein. Science 250:1587–1590
    [Google Scholar]
  28. Kascsak R. J., Rubenstein R., Merz P. A., Carp R. I., Robakis N. K., Wisniewski H. M., Diringer H. 1986; Immunological comparison of scrapie-associated fibrils isolated from animals infected with four different scrapie strains. Journal of Virology 59:676–683
    [Google Scholar]
  29. Kascsak R. J., Rubenstein R., Merz P. A., Tonna-DeMasi M., Fersko R., Carp R. I., Wisniewski H. M., Diringer H. 1987; Mouse polyclonal and monoclonal antibody to scrapie-associated fibril proteins. Journal of Virology 61:3688–3693
    [Google Scholar]
  30. Kimberlin R. H. 1979; The biology of scrapie agent. In Aspects of Slow and Persistent Virus Infections pp 4–29 Edited by Tyrrell D. A. J. The Hague: Martinus Nijhoff;
    [Google Scholar]
  31. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  32. Leary J. J., Brigate D. J., Ward D. C. 1983; Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose bio-blots. Proceedings of the National Academy of Sciences, U.S.A. 80:4045–4049
    [Google Scholar]
  33. McKinley M. P., Bolton D. C., Prusiner S. B. 1983; A protease-resistant protein is a structural component of the scrapie prion. Cell 35:57–62
    [Google Scholar]
  34. McKinley M. P., Braunfeld M. B., Bellinger C. G., Prusiner S. B. 1986; Molecular characteristics of prion rods purified from scrapie-infected hamster brains. Journal of Infectious Diseases 154:110–120
    [Google Scholar]
  35. McKinley M. P., Hay B., Lingappa V. R., Lieberburg I., Prusiner S. B. 1987; Developmental expression of prion protein gene in brain. Developmental Biology 121:105–110
    [Google Scholar]
  36. Manuelidis L., Sklaviadis T., Manuelidis E. E. 1987; Evidence suggesting that PrP is not the infectious agent in Creutzfeldt-Jakob disease. EMBO Journal 6:341–347
    [Google Scholar]
  37. Marsh R. F., Castle B. E., Dees C., Wade W. F. 1984a; Equilibrium density gradient centrifugation of the scrapie agent in Nycodenz. Journal of General Virology 65:1963–1968
    [Google Scholar]
  38. Marsh R. F., Dees C., Castle B. E., Wade W. F., German T. L. 1984b; Purification of the scrapie agent by density gradient centrifugation. Journal of General Virology 65:415–421
    [Google Scholar]
  39. Masters C. L., Rohwer R. G., Franko M. C., Brown P., Gajdusek D. C. 1984; The sequential development of spongiform change and gliosis of scrapie in the golden Syrian hamster. Journal of Neuropathology and Experimental Neurology 43:242–252
    [Google Scholar]
  40. Merz P. A., Somerville R. A., Wisniewski H. M., Iqbal K. 1981; Abnormal fibils from scrapie-infected brain. Acta neuropathologica 54:63–74
    [Google Scholar]
  41. Merz P. A., Somerville R. A., Wisniewski H. M., Manuelidis L., Manuelidis E. E. 1983; Scrapie-associated fibrils in Creutzfeldt-Jakob disease. Nature, London 306:474–476
    [Google Scholar]
  42. Merz P. A., Rohwer R. G., Kascsak R. J., Wisniewski H. M., Somerville R. A., Gibbs C. J. Jr, Gajdusek D. C. 1984; Infection-specific particle from the unconventional slow virus diseases. Science 225:437–440
    [Google Scholar]
  43. Meyer R. K., McKinley M. P., Bowman K. A., Braunfeld M. B., Barry R. A., Prusiner S. B. 1986; Separation and properties of cellular and scrapie prion proteins. Proceedings of the National Academy of Sciences, U.S.A. 83:2310–2314
    [Google Scholar]
  44. Murphy F. A., Kingsbury D. W. 1990; Virus taxonomy. In Virology 2nd edn pp 9–35 Edited by Fields B. N., Knipe D. M. New York: Raven Press;
    [Google Scholar]
  45. Oesch B., Groth D. F., Prusiner S. B., Weissmann C. 1988; Search for a scrapie-specific nucleic acid: a progress report. Ciba Foundation Symposium 135:209–223
    [Google Scholar]
  46. Prusiner S. B. 1982; Novel proteinaceous infectious particles cause scrapie. Science 216:136–144
    [Google Scholar]
  47. Prusiner S. B. 1987; The prion hypothesis. In Prions: Novel Infectious Pathogens Causing Scrapie and Creutzfeldt-Jakob Disease pp 17–36 Edited by Prusiner S. B., McKinley M. P. San Diego: Academic Press;
    [Google Scholar]
  48. Prusiner S. B., McKinley M. P. (editors) 1987 Prions: Novel Infectious Pathogens Causing Scrapie and Creutzfeldt-Jakob Disease San Diego: Academic Press;
    [Google Scholar]
  49. Prusiner S. B., Bolton D. C., Groth D. F., Bowman K. A., Cochran S. P., McKinley M. P. 1982a; Further purification and characterization of scrapie prions. Biochemistry 21:6942–6950
    [Google Scholar]
  50. Prusiner S. B., Cochran S. P., Groth D. F., Downey D. E., Bowman K. A., Martinez H. M. 1982b; Measurement of the scrapie agent using an incubation time interval assay. Annals of Neurology 11:353–358
    [Google Scholar]
  51. Prusiner S. B., McKinley M. P., Bowman K. A., Bolton D. C., Bendheim P. E., Groth D. F., Glenner G. G. 1983; Scrapie prions aggregate to form amyloid-like birefringent rods. Cell 35:349–358
    [Google Scholar]
  52. Safar J., Wang W., Padgett M. P., Ceroni M., Piccardo P., Zopf D., Gajdusek D. C., Gibbs C. J. Jr 1990; Molecular mass, biochemical composition, and physicochemical behavior of the infectious form of the scrapie precursor protein monomer. Proceedings of the National Academy of Sciences, U.S.A. 87:6373–6377
    [Google Scholar]
  53. Scott M., Foster D., Mirenda C. A., Serban D., Coufal F., Walchli M., Torchia M., Groth D., Carlson G. A., DeArmond S. J., Westaway D., Prusiner S. B. 1989; Transgenic mice expressing hamster prion protein produce species-specific scrapie infectivity and amyloid plaques. Cell 59:847–857
    [Google Scholar]
  54. Sklaviadis T., Manuelidis L., Manuelidis E. E. 1986; Characterization of major peptides in Creutzfeldt-Jakob disease and scrapie. Proceedings of the National Academy of Sciences, U.S.A. 83:6146–6150
    [Google Scholar]
  55. Sklaviadis T. K., Manuelidis L., Manuelidis E. E. 1989; Physical properties of the Creutzfeldt-Jakob disease agent. Journal of Virology 63:1212–1222
    [Google Scholar]
  56. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A. 76:4350–4354
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-12-2905
Loading
/content/journal/jgv/10.1099/0022-1317-72-12-2905
Loading

Data & Media loading...

Most cited Most Cited RSS feed