Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 May;34(5):1228–1231. doi: 10.1128/jcm.34.5.1228-1231.1996

Immunohistochemical detection of prion protein in lymphoid tissues of sheep with natural scrapie.

L J van Keulen 1, B E Schreuder 1, R H Meloen 1, G Mooij-Harkes 1, M E Vromans 1, J P Langeveld 1
PMCID: PMC228987  PMID: 8727908

Abstract

The scrapie-associated form of the prion protein (PrPSc) accumulates in the brain and lymphoid tissues of sheep with scrapie. In order to assess whether detecting PrPSc in lymphoid tissue could be used as a diagnostic test for scrapie, we studied the localization and distribution of PrPSc in various lymphoid tissues collected at necropsy from 55 sheep with clinical scrapie. Samples collected from the spleen, palatine tonsil, ileum, and five different lymph nodes were immunohistochemically stained for PrPSc. PrPSc was found to be deposited in a reticular pattern in the center of both primary and secondary lymphoid follicles. In addition, granules of PrPSc were seen in the cytoplasm in macrophages associated with the lymphoid follicles. In 54 (98%) of the 55 scrapie-affected sheep, PrPSc was detected in the spleen, retropharyngeal lymph node, mesenteric lymph node, and the palatine tonsil. However, only in the palatine tonsils was PrPSc present in a consistently high percentage of the lymphoid follicles. PrP was not detected in any of the lymphoid tissues of 12 sheep that had no neurohistopathological signs of a scrapie infection. We conclude that the tonsils are the best-suited lymphoid tissue to be biopsied for the detection of PrPSc in the diagnosis of clinical scrapie in living sheep.

Full Text

The Full Text of this article is available as a PDF (822.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Basler K., Oesch B., Scott M., Westaway D., Wälchli M., Groth D. F., McKinley M. P., Prusiner S. B., Weissmann C. Scrapie and cellular PrP isoforms are encoded by the same chromosomal gene. Cell. 1986 Aug 1;46(3):417–428. doi: 10.1016/0092-8674(86)90662-8. [DOI] [PubMed] [Google Scholar]
  2. Belt P. B., Muileman I. H., Schreuder B. E., Bos-de Ruijter J., Gielkens A. L., Smits M. A. Identification of five allelic variants of the sheep PrP gene and their association with natural scrapie. J Gen Virol. 1995 Mar;76(Pt 3):509–517. doi: 10.1099/0022-1317-76-3-509. [DOI] [PubMed] [Google Scholar]
  3. Diringer H., Gelderblom H., Hilmert H., Ozel M., Edelbluth C., Kimberlin R. H. Scrapie infectivity, fibrils and low molecular weight protein. Nature. 1983 Dec 1;306(5942):476–478. doi: 10.1038/306476a0. [DOI] [PubMed] [Google Scholar]
  4. Doi S., Ito M., Shinagawa M., Sato G., Isomura H., Goto H. Western blot detection of scrapie-associated fibril protein in tissues outside the central nervous system from preclinical scrapie-infected mice. J Gen Virol. 1988 Apr;69(Pt 4):955–960. doi: 10.1099/0022-1317-69-4-955. [DOI] [PubMed] [Google Scholar]
  5. Eklund C. M., Kennedy R. C., Hadlow W. J. Pathogenesis of scrapie virus infection in the mouse. J Infect Dis. 1967 Feb;117(1):15–22. doi: 10.1093/infdis/117.1.15. [DOI] [PubMed] [Google Scholar]
  6. Fraser H., Dickinson A. G. Pathogenesis of scrapie in the mouse: the role of the spleen. Nature. 1970 May 2;226(5244):462–463. doi: 10.1038/226462a0. [DOI] [PubMed] [Google Scholar]
  7. Fraser H., Dickinson A. G. Studies of the lymphoreticular system in the pathogenesis of scrapie: the role of spleen and thymus. J Comp Pathol. 1978 Oct;88(4):563–573. doi: 10.1016/0021-9975(78)90010-5. [DOI] [PubMed] [Google Scholar]
  8. Hadlow W. J., Kennedy R. C., Race R. E. Natural infection of Suffolk sheep with scrapie virus. J Infect Dis. 1982 Nov;146(5):657–664. doi: 10.1093/infdis/146.5.657. [DOI] [PubMed] [Google Scholar]
  9. Hilmert H., Diringer H. A rapid and efficient method to enrich SAF-protein from scrapie brains of hamsters. Biosci Rep. 1984 Feb;4(2):165–170. doi: 10.1007/BF01120313. [DOI] [PubMed] [Google Scholar]
  10. Ikegami Y., Ito M., Isomura H., Momotani E., Sasaki K., Muramatsu Y., Ishiguro N., Shinagawa M. Pre-clinical and clinical diagnosis of scrapie by detection of PrP protein in tissues of sheep. Vet Rec. 1991 Mar 23;128(12):271–275. doi: 10.1136/vr.128.12.271. [DOI] [PubMed] [Google Scholar]
  11. Kitamoto T., Mohri S., Tateishi J. Organ distribution of proteinase-resistant prion protein in humans and mice with Creutzfeldt-Jakob disease. J Gen Virol. 1989 Dec;70(Pt 12):3371–3379. doi: 10.1099/0022-1317-70-12-3371. [DOI] [PubMed] [Google Scholar]
  12. Kitamoto T., Muramoto T., Mohri S., Doh-Ura K., Tateishi J. Abnormal isoform of prion protein accumulates in follicular dendritic cells in mice with Creutzfeldt-Jakob disease. J Virol. 1991 Nov;65(11):6292–6295. doi: 10.1128/jvi.65.11.6292-6295.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McBride P. A., Eikelenboom P., Kraal G., Fraser H., Bruce M. E. PrP protein is associated with follicular dendritic cells of spleens and lymph nodes in uninfected and scrapie-infected mice. J Pathol. 1992 Dec;168(4):413–418. doi: 10.1002/path.1711680412. [DOI] [PubMed] [Google Scholar]
  14. McKinley M. P., Bolton D. C., Prusiner S. B. A protease-resistant protein is a structural component of the scrapie prion. Cell. 1983 Nov;35(1):57–62. doi: 10.1016/0092-8674(83)90207-6. [DOI] [PubMed] [Google Scholar]
  15. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  16. Miller J. M., Jenny A. L., Taylor W. D., Marsh R. F., Rubenstein R., Race R. E. Immunohistochemical detection of prion protein in sheep with scrapie. J Vet Diagn Invest. 1993 Jul;5(3):309–316. doi: 10.1177/104063879300500301. [DOI] [PubMed] [Google Scholar]
  17. Mohri S., Farquhar C. F., Somerville R. A., Jeffrey M., Foster J., Hope J. Immunodetection of a disease specific PrP fraction in scrapie-affected sheep and BSE-affected cattle. Vet Rec. 1992 Dec 5;131(23):537–539. [PubMed] [Google Scholar]
  18. Muramatsu Y., Onodera A., Horiuchi M., Ishiguro N., Shinagawa M. Detection of PrPSc in sheep at the preclinical stage of scrapie and its significance for diagnosis of insidious infection. Arch Virol. 1994;134(3-4):427–432. doi: 10.1007/BF01310580. [DOI] [PubMed] [Google Scholar]
  19. Muramoto T., Kitamoto T., Tateishi J., Goto I. Accumulation of abnormal prion protein in mice infected with Creutzfeldt-Jakob disease via intraperitoneal route: a sequential study. Am J Pathol. 1993 Nov;143(5):1470–1479. [PMC free article] [PubMed] [Google Scholar]
  20. Race R. E., Ernst D. Detection of proteinase K-resistant prion protein and infectivity in mouse spleen by 2 weeks after scrapie agent inoculation. J Gen Virol. 1992 Dec;73(Pt 12):3319–3323. doi: 10.1099/0022-1317-73-12-3319. [DOI] [PubMed] [Google Scholar]
  21. Race R., Ernst D., Jenny A., Taylor W., Sutton D., Caughey B. Diagnostic implications of detection of proteinase K-resistant protein in spleen, lymph nodes, and brain of sheep. Am J Vet Res. 1992 Jun;53(6):883–889. [PubMed] [Google Scholar]
  22. Rubenstein R., Kascsak R. J., Merz P. A., Papini M. C., Carp R. I., Robakis N. K., Wisniewski H. M. Detection of scrapie-associated fibril (SAF) proteins using anti-SAF antibody in non-purified tissue preparations. J Gen Virol. 1986 Apr;67(Pt 4):671–681. doi: 10.1099/0022-1317-67-4-671. [DOI] [PubMed] [Google Scholar]
  23. Rubenstein R., Merz P. A., Kascsak R. J., Carp R. I., Scalici C. L., Fama C. L., Wisniewski H. M. Detection of scrapie-associated fibrils (SAF) and SAF proteins from scrapie-affected sheep. J Infect Dis. 1987 Jul;156(1):36–42. doi: 10.1093/infdis/156.1.36. [DOI] [PubMed] [Google Scholar]
  24. Shinagawa M., Munekata E., Doi S., Takahashi K., Goto H., Sato G. Immunoreactivity of a synthetic pentadecapeptide corresponding to the N-terminal region of the scrapie prion protein. J Gen Virol. 1986 Aug;67(Pt 8):1745–1750. doi: 10.1099/0022-1317-67-8-1745. [DOI] [PubMed] [Google Scholar]
  25. Somerville R. A., Merz P. A., Carp R. I. Partial copurification of scrapie-associated fibrils and scrapie infectivity. Intervirology. 1986;25(1):48–55. doi: 10.1159/000149654. [DOI] [PubMed] [Google Scholar]
  26. van Keulen L. J., Schreuder B. E., Meloen R. H., Poelen-van den Berg M., Mooij-Harkes G., Vromans M. E., Langeveld J. P. Immunohistochemical detection and localization of prion protein in brain tissue of sheep with natural scrapie. Vet Pathol. 1995 May;32(3):299–308. doi: 10.1177/030098589503200312. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES