Cellular and sub-cellular localisation of PrP in the lymphoreticular system of mice and sheep

  • M. Jeffrey
  • G. McGovern
  • S. Martin
  • C. M. Goodsir
  • K. L. Brown
Part of the Archives of Virology. Supplementa book series (ARCHIVES SUPPL, volume 16)


Using immunocytochemistry or immunogold electron microscopy, abnormal PrP accumulation was found in lymphoreticular tissues of Suffolk sheep naturally exposed to scrapie and in the spleens of ME7 infected C57 BL mice at 70 days after infection and at the terminal stage of disease at 170 days. Clinically diseased scrapie affected sheep show widespread PrP accumulation within tingible body macrophages (TBMs) and follicular dendritic cells (FDCs) of secondary lymphoid follicles. Serial tonsillar biopsies taken from 171ARQ/ARQ sheep at 4 months of age did not contain abnormal PrP accumulations but 80% of biopsies were positive by 14 months. In contrast, whole body necropsies of sheep not previously biopsied failed to detect PrP in the tonsil of sheep at 4, 8, 12 or 16 months of age. These findings suggest that the biopsy procedure of susceptible sheep but not resistant sheep may induce tonsillar infection. In spleen of mice both at 70 and 170 dpi, accumulations of PrP were found within lysosomes of TBMs and also at the plasma-lemma of FDCs. In the light zone of follicles of terminally diseased mice, all FDC dendrites were arranged in the form of highly reactive or hyperplastic labrynthine glomerular complexes. PrP was consistently seen between FDC dendrites in association with abundant electron dense antigen-antibody complexes. At 70 days after challenge, labrynthine complexes were rare and invariably labelled for PrP. However, sparse PrP labelling was also seen on simple FDC dendrites at this stage. These observations suggests that scrapie infected FDCs continually release PrP from the cell surface where it accumulates in excess in association with trapped immune complexes and dendritic extension. It is likely that TBMs acquire lysosomal PrP following phagocytosis of effete FDC processes or from the extracellular space. We suggest that the normal function of PrP may involve cell process extension or immune complex trapping.


Germinal Centre Follicular Dendritic Cell Electron Dense Deposit Light Zone Murine Spleen 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    Blattler T, Brandner S, Raeber AJ, Klein MA, Voigtlander T, Weissmann C, Aguzzi A (1997) PrP-expressing tissue required for transfer of scrapie infectivity from spleen to brain. Nature 389: 69–73PubMedCrossRefGoogle Scholar
  2. 2.
    Bosma GC, Fried M, Custer RP, Carroll A, Gibson DM, Bosma MJ (1983) A severe combined immunodeficiency mutation in the mouse. Nature 301: 527–530PubMedCrossRefGoogle Scholar
  3. 3.
    Bosseloir A, Bouzahzah F, Simar L, Heinen E (1995) B cells in contact with FDC. In: Heinen E (ed) Follicular dendritic cells in normal and pathological conditions. Springer, Berlin Heidelberg New York Tokyo, pp 53–78Google Scholar
  4. 4.
    Brown KL, Stewart K, Bruce ME, Fraser H (1997) Severely combined immunodeficient (SCID) mice resist infection with bovine spongiform encephalo-pathy. J Gen Virol 78: 2707–2710PubMedGoogle Scholar
  5. 5.
    Brown KL, Stewart K, Ritchie D, Mabbott NA, Williams A, Fraser H, Morrison WI, Bruce ME (2000) Scrapie replication in lymphoid tissues depends on prion PrP-expressing follicular dendritic cells. Nature Med 11: 1308–1311Google Scholar
  6. 6.
    Brown KL, Stewart K, Bruce ME, Fraser H (1996) Scrapie in immunodeficient mice. In: Court L, Dodet B (eds) Transmissible subacute spongiform encephalo-pathies. Elsevier, Paris, pp 159–166Google Scholar
  7. 7.
    Carp RI, Callaghan SM, Patrick BA, Mehta PD (1994) Interaction of scrapie agent and cells of the lymphoreticular system. Arch Virol 136: 255–268PubMedCrossRefGoogle Scholar
  8. 8.
    Cashman NR, Loertscher R, Nalbantoglu J (1990) Cellular isoform of the scrapie agent protein participates in lymphocyte activation. Cell 61: 185PubMedCrossRefGoogle Scholar
  9. 9.
    Collis CS, Kimberlin RH (1985) Long term persistence of scrapie infection in mouse spleens in the absence of clinical disease. FEMS Microbiol Lett 29: 111–114CrossRefGoogle Scholar
  10. 10.
    Collis SC, Kimberlin RF (1983) Further studies on changes in immunoglobulin G in the sera and CSF of Herdwick sheep with natural and experimental scrapie. J Comp Pathol 93: 331–338PubMedCrossRefGoogle Scholar
  11. 11.
    Diringer H, Gelderblom H, Hilmert H, Ozel M, Edelbluth C (1983) Scrapie infectivity, fibrils and low molecular weight protein. Nature 306: 476–478PubMedCrossRefGoogle Scholar
  12. 12.
    Fraser H, Dickinson AG (1978) Studies of lymphoreticular system in the pathogenesis of scrapie. The role of spleen and thymus. J Comp Pathol 88: 563–573PubMedCrossRefGoogle Scholar
  13. 13.
    Fraser H, Farquhar C (1987) Ionising radiation has no influence on scrapie incubation period in mice. Vet Microbiol 13: 211–223PubMedCrossRefGoogle Scholar
  14. 14.
    Harris DA (1999) Cell biological studies of the prion protein. In: Harris DA (ed) Prions: molecular and cellular biology. Horizon Scientific Press, Norfolk, pp 53–66Google Scholar
  15. 15.
    Heinen E, Bosseloir A, Bouzahzah F (1995) Follicular dendritic cells: origin and function. Curr Top Microbiol Immunol 201: 15–47PubMedCrossRefGoogle Scholar
  16. 16.
    Hill AF, Zeidler M, Ironside J, Collinge J (1997) Diagnosis of new variant Creutzfeldt-Jakob disease by tonsil biopsy. Lancet 349: 99–100PubMedCrossRefGoogle Scholar
  17. 17.
    Jeffrey M, Goodsir CM, Bruce ME, McBride PA, Scott JR, Halliday WG (1994) Correlative light and electron microscopy studies of PrP localisation in 87V scrapie. Brain Res 656: 329–343PubMedCrossRefGoogle Scholar
  18. 18.
    Jeffrey M, Goodsir CM, Bruce ME, McBride PA, Fowler N, Scott JR (1994) Murine scrapie-infected neurons in vivo release excess PrP into the extracellular space. Neurosci Lett 174: 39–42PubMedCrossRefGoogle Scholar
  19. 19.
    Jeffrey M, Goodsir CM, Fowler N, Hope J, Bruce ME, McBride PA (1996) Ultrastructural immunolocalisation of synthetic prion protein peptide antibodies in 87V murine scrapie. Neurodegeneration 5: 101–109PubMedCrossRefGoogle Scholar
  20. 20.
    Jeffrey M, McGovern G, Goodsir CM, Brown KL, Bruce ME (2000) Sites of prion protein accumulation in scrapie-infectd mouse spleen revealed by immuno-electron microscopy. J Pathol (in press)Google Scholar
  21. 21.
    Kimberlin RH, Walker CA (1988) Incubation periods in six models of intraperitoneally injected scrapie depend mainly on the dynamics of agent replication within the nervous system and not the lymphoreticular system. J Gen Virol 69: 2953–2960PubMedCrossRefGoogle Scholar
  22. 22.
    Kitamoto T, Muramoto T, Mohri S, Doh-ura K, Tatieishi J (1991) Abnormal isoform of prion protein accumulates in follicular dendritic cells in mice with Creutzfeldt-Jakob Disease. J Virol 65: 6292–6295PubMedGoogle Scholar
  23. 23.
    Klein MA, Frigg R, Raeber AJ, Flechsig E, Hegyi I, Zinkernagel RM, Weissmann C, Aguzzi A (1998) PrP expresssion in B lymphocytes is not required for prion invasion. Nature Med 4: 1429–1433PubMedCrossRefGoogle Scholar
  24. 24.
    Laszemas CI, Cesbron J-Y, Deslys J-P, Demaimay R, Adjou KT, Rioux R, Lemaire C, Locht C, Dormont D (1996) Immune system-dependent and independent replication of the scrapie agent. J Virol 70: 1292–1295Google Scholar
  25. 25.
    Mabbott NA, Brown KL, Manson J, Bruce ME (1997) T lymphocyte activation and the cellular form of the prion protein. Immunology 92: 161–165PubMedCrossRefGoogle Scholar
  26. 26.
    Manson J, McBride P, Hope J (1992) Expression of the PrP gene in the brain of sinc congenic mice and its relationship to the development of scrapie. Neurodegeneration 1: 45–52Google Scholar
  27. 27.
    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. Pathol 168: 413–418CrossRefGoogle Scholar
  28. 28.
    Ritchie DL, Brown KL, Bruce ME (2000) Visualisation of PrP protein and follicular dendritic cells in uninfected and scrapie infected spleen. J Cell Pathol 1: 3–10Google Scholar
  29. 29.
    Shyng S-L, Heuser JE, Harris DA (1994) A glycolipid-anchored prion protein is endocytosed via clathrin coate pits. J Cell Biol 124: 1239–1250CrossRefGoogle Scholar
  30. 30.
    Szakal AK, Kosco MH, Tew JG (1988) A novel in vivo follicular dendritic cell-dependent iccosome-mediated mechanism for delivery of antigen to antigen-processing cells. J Immunol 140: 342–353Google Scholar
  31. 31.
    Terashima K, Dobashi M, Maeda K, Imai Y (1992). Follicular dendritic cell and iccosomes in germinal centre reactions. Semin Immunol 4: 267–274PubMedGoogle Scholar
  32. 32.
    Tew JG, Kosco MH, Szakal AK (1989) The alternative antigen pathway. Immunol Today 10: 229–231PubMedCrossRefGoogle Scholar
  33. 33.
    van Keulen LJM, Schreuder BEC, Meloen RH, MooijHarkes G, Vromans MEW, Langeveld JPM (1996) Immunohistochemical detection of prion protein in lymphoid tissues of sheep with natural scrapie. J Clin Microbiol 34: 1228–1231PubMedGoogle Scholar
  34. 34.
    Schreuder BEC, van Keulen LJM, Vromans MEW, Langeveld JPM, Smits M (1998) Tonsillar biopsy and PrPSc detection in the preclinical diagnosis of scrapie. Vet Rec 142: 564–568PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2000

Authors and Affiliations

  • M. Jeffrey
    • 1
  • G. McGovern
    • 1
  • S. Martin
    • 1
  • C. M. Goodsir
    • 1
  • K. L. Brown
    • 1
  1. 1.Lasswade Veterinary LaboratoryPenicuikScotland, UK

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