Springer Protocols: Abstract: 5. Generation of Cell Lines Propagating Infectious Prions and the Isolation and Characterization of Cell-derived Exosomes

5. Generation of Cell Lines Propagating Infectious Prions and the Isolation and Characterization of Cell-derived Exosomes

By: Laura J. Vella³, Andrew F. Hill³

Abstract

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Prion-propagating cell lines are an efficient and useful means for studying the cellular and molecular mechanisms implicated in prion disease. Use of cell-based models has lead to the finding that prion protein (PrP^C) and PrP^Sc are released from cells in association with exosomes. Furthermore, exosomes have been shown to act as vehicles for infectivity, transferring PrP^Sc between cell lines and providing a mechanism for prion spread between tissues. As a role for exo-somes in prion disease is emerging, this chapter outlines a method for the generation of prion-infected cell lines and the isolation and characterization of PrP^C- and PrP^Sc-containing exosomes.

Affiliation(s): (3) Department of Biochemistry and Molecular Biology and Department of Pathology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Melbourne, Australia

Book Title: Prion Protein Protocols

Series: Methods in Molecular Biology | Volume: 459 | Pub. Date: Jun-04-2008 | Page Range: 69-82 | DOI: 10.1007/978-1-59745-234-2_5

Subject: Protein Science

Key Words: Cell culture media - exosomes - GT1–7 - infection - passaging - prion

References

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1.	Prusiner, S.B. (1982) Novel proteinaceous infectious particles cause scrapie. Science 216(4542): 136–44.

2.	Clarke, M.C. and D.A. Haig. (1970) Evidence for the multiplication of scrapie agent in cell culture. Nature 225(5227): 100–1.

3.	Race, R.E., L.H. Fadness, and B. Chesebro. (1987) Characterization of scrapie infection in mouse neuroblastoma cells. J Gen Virol 68(5): 1391–9.

4.	Bosque, P.J. and S.B. Prusiner. (2000) Cultured cell sublines highly susceptible to prion infection. J Virol 74(9): 4377–86.

5.	Schatzl, H.M., et al. (1997) A hypothalamic neuronal cell line persistently infected with scrapie prions exhibits apoptosis. J Virol 71(11): 8821–31.

6.	Vorberg, I., et al. (2004) Susceptibility of common fibroblast cell lines to transmissible spong-iform encephalopathy agents. J Infect Dis 189: 431–9.

7.	Priola, S.A., et al. (1994) Prion protein and the scrapie agent: in vitro studies in infected neu-roblastoma cells. Infect Agents Dis 3(2–3): 54–8.

8.	Supattapone, S., et al. (2001) Branched polyamines cure prion-infected neuroblastoma cells. J Virol 75(7): 3453–61.

9.	Fevrier, B., et al. (2004) Cells release prions in association with exosomes. Proc Natl Acad Sci U S A 101(26): 9683–8.

10.	Stoorvogel, W., et al. (2002) The biogenesis and functions of exosomes. Traffic 3(5): 321–30.

11.	Denzer, K., et al. (2000) Follicular dendritic cells carry MHC class II-expressing microvesi-cles at their surface. J Immunol 165(3): 1259–65.

12.	Pisitkun, T., R.F. Shen, and M.A. Knepper. (2004) Identification and proteomic profiling of exosomes in human urine. Proc Natl Acad Sci U S A 101(36): 13368–73.

13.	Andre, F., et al. (2002) Tumor-derived exosomes: a new source of tumor rejection antigens. Vaccine 20(Suppl 4): A28–31.

14.	Gastpar, R., et al. (2005) Heat shock protein 70 surface-positive tumor exosomes stimulate migratory and cytolytic activity of natural killer cells. Cancer Res 65(12): 5238–47.

15.	Abusamra, A.J., et al. (2005) Tumor exosomes expressing Fas ligand mediate CD8+ T-cell apoptosis. Blood Cells Mol Dis 35(2): 169–73.

16.	Cho, J.A., et al. (2004) Exosomes: a new delivery system for tumor antigens in cancer immu-notherapy. Int J Cancer 114(4): 613–22.

17.	Hee Kim, S., et al. (2005) Exosomes derived from genetically modified DC expressing FasL are anti-inflammatory and immunosuppressive. Mol Ther 13(2): 289–300.

18.	Skokos, D., et al. (2001) Mast cell-dependent B and T lymphocyte activation is mediated by the secretion of immunologically active exosomes. J Immunol 166(2): 868–76.

19.	Wolfers, J., et al. (2001) Tumor-derived exosomes are a source of shared tumor rejection antigens for CTL cross-priming. Nat Med 7(3): 297–303.

20.	Skokos, D., et al. (2003) Mast cell-derived exosomes induce phenotypic and functional maturation of dendritic cells and elicit specific immune responses in vivo. J Immunol 170(6): 3037–45.

21.	Van Niel, G., et al. (2003) Intestinal epithelial exosomes carry MHC class II/peptides able to inform the immune system in mice. Gut 52(12): 1690–7.

22.	Kim, S.H., et al. (2005) Exosomes derived from IL-10-treated dendritic cells can suppress inflammation and collagen-induced arthritis. J Immunol 174(10): 6440–8.

23.	Wiley, R.D. and S. Gummuluru. (2006) Immature dendritic cell-derived exosomes can mediate HIV-1 trans infection. Proc Natl Acad Sci U S A 103(3): 738–43.

24.	Ecroyd, H., et al. (2006) An epididymal form of cauxin, a carboxylesterase-like enzyme, is present and active in mammalian male reproductive fluids. Biol Reprod 74(2): 439–447.

25.	Ecroyd, H., et al. (2004) Compartmentalization of prion isoforms within the reproductive tract of the ram. Biol Reprod 71(3): 993–1001.

26.	Gatti, J.L., et al. (2002) Prion protein is secreted in soluble forms in the epididymal fluid and proteolytically processed and transported in seminal plasma. Biol Reprod 67(2): 393–400.

27.	Robertson, C., et al. (2006) Cellular prion protein is released on exosomes from activated platelets. Blood 107(10): 3907–11.

28.	Faure, J., et al. (2006) Exosomes are released by cultured cortical neurones. Mol Cell Neurosci 31(4): 642–8.

29.	Vella, L.J., et al. (2007) Packaging of prions into exosomes is associated with a novel pathway of PrP processing. J Pathol 211(5): 582–90.

30.	Aguzzi, A. (2003) Prions and the immune system: a journey through gut, spleen, and nerves. Adv Immunol 81: 123–71.

31.	Brown, K.L., et al. (1999) Scrapie replication in lymphoid tissues depends on prion protein-expressing follicular dendritic cells. Nat Med 5(11): 1308–12.

32.	Kitamoto, T., et al. (1991) Abnormal isoform of prion protein accumulates in follicular dendritic cells in mice with Creutzfeldt-Jakob disease. J Virol 65(11): 6292–5.

33.	Andreoletti, O., et al. (2000) Early accumulation of PrP(Sc) in gut-associated lymphoid and nervous tissues of susceptible sheep from a Romanov flock with natural scrapie. J Gen Virol 81(12): 3115–26.

34.	van Keulen, L.J., et al. (2000) Pathogenesis of natural scrapie in sheep. Arch Virol Suppl (16): 57–71.

35.	McBride, P.A., et al. (1992) PrP protein is associated with follicular dendritic cells of spleens and lymph nodes in uninfected and scrapie-infected mice. J Pathol 168(4): 413–8.

36.	Clarke, M.C. and R.H. Kimberlin. (1984) Pathogenesis of mouse scrapie: distribution of agent in the pulp and stroma of infected spleens. Vet Microbiol 9(3): 215–25.

37.	Kimberlin, R.H. and C.A. Walker. (1989) Pathogenesis of scrapie in mice after intragastric infection. Virus Res 12(3): 213–20.

38.	Beekes, M. and P.A. McBride. (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(3): 181–4.

39.	Baldauf, E., M. Beekes, and H. Diringer. (1997) Evidence for an alternative direct route of access for the scrapie agent to the brain bypassing the spinal cord. J Gen Virol 78(5): 1187–97.

40.	Butler, D.A., et al. (1988) Scrapie-infected murine neuroblastoma cells produce protease-resistant prion proteins. J Virol 62(5): 1558–64.

41.	Rubenstein, R., R.I. Carp, and S.M. Callahan. (1984) In vitro replication of scrapie agent in a neuronal model: infection of PC12 cells. J Gen Virol 65(12): 2191–8.

42.	Vilette, D., et al. (2001) Ex vivo propagation of infectious sheep scrapie agent in heterologous epithelial cells expressing ovine prion protein. Proc Natl Acad Sci U S A 98(7): 4055–9.

43.	Clarke, M.C. and G.C. Millson. (1976) Infection of a cell line of mouse L fibroblasts with scrapie agent. Nature 261(5556): 144–5.

44.	Gibson, P.E., T.M. Bell, and E.J. Field. (1972) Failure of the scrapie agent to replicate in L5178Y mouse leukaemic cells. Res Vet Sci 13(1): 95–6.

45.	Nishida, N., et al. (2000) Successful transmission of three mouse-adapted scrapie strains to murine neuroblastoma cell lines overexpressing wild-type mouse prion protein. J Virol 74(1): 320–5.

46.	Arjona, A., et al. (2004) Two Creutzfeldt-Jakob disease agents reproduce prion protein-independent identities in cell cultures. Proc Natl Acad Sci U S A 101(23): 8768–73.

47.	Raposo, G., et al. (1996) B lymphocytes secrete antigen-presenting vesicles. J Exp Med 183(3): 1161–72.

48.	Thery, C., et al. (1999) Molecular characterization of dendritic cell-derived exosomes. Selective accumulation of the heat shock protein hsc73. J Cell Biol 147(3): 599–610.

49.	Clayton, A., et al. (2001) Analysis of antigen presenting cell derived exosomes, based on immuno-magnetic isolation and flow cytometry. J Immunol Methods 247(1–2): 163–74.

50.	Cheruvanky, A., et al. (2007) Rapid isolation of urinary exosomal biomarkers using a nanomembrane ultrafiltration concentrator. Am J Physiol Renal Physiol 292(5): F1657–61.

51.	van Niel, G. and M. Heyman. (2002) The epithelial cell cytoskeleton and intracellular trafficking. II. Intestinal epithelial cell exosomes: perspectives on their structure and function. Am J Physiol Gastrointest Liver Physiol 283(2): G251–5.

52.	Thery, C., et al. (2001) Proteomic analysis of dendritic cell-derived exosomes: a secreted subcellular compartment distinct from apoptotic vesicles. J Immunol 166(12): 7309–18.

53.	Hegmans, J.P., et al. (2004) Proteomic analysis of exosomes secreted by human mesothelioma cells. Am J Pathol 164(5): 1807–15.

54.	Mears, R., et al. (2004) Proteomic analysis of melanoma-derived exosomes by two-dimensional polyacrylamide gel electrophoresis and mass spectrometry. Proteomics 4(12): 4019–31.

55.	Bard, M.P., et al. (2004) Proteomic analysis of exosomes isolated from human malignant pleural effusions. Am J Respir Cell Mol Biol 31(1): 114–21.

56.	Wubbolts, R., et al. (2003) Proteomic and biochemical analyses of human B cell-derived exo-somes. Potential implications for their function and multivesicular body formation. J Biol Chem 278(13): 10963–72.

57.	Segura, E., S. Amigorena, and C. Thery. (2005) Mature dendritic cells secrete exosomes with strong ability to induce antigen-specific effector immune responses. Blood Cells Mol Dis 35(2): 89–93.

58.	Amzallag, N., et al. (2004) TSAP6 facilitates the secretion of translationally controlled tumor pro-tein/histamine-releasing factor via a nonclassical pathway. J Biol Chem 279(44): 46104–12.

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