In most forms of prion diseases, blood is infectious, but detection by immunochemistry techniques of the only available marker of infection (the misfolded prion protein, PrP) in blood remains elusive. We developed a novel method for the detection of PrP in blood of prion-infected rodents based on the finding that PrP is associated with plasma exosomes. However, further purification of the exosomes on a sucrose gradient was necessary to remove plasma immunoglobulins, which interfere with PrP, masking its detection by immunochemistry. Finally, we report that about 20% of plasma infectivity is associated with exosomes.


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  1. Abdel-Haq H. (2015). Factors intrinsic and extrinsic to blood hamper the development of a routine blood test for human prion diseasesJ Gen Virol 96479493 [View Article][PubMed]. [Google Scholar]
  2. Alais S., Simoes S., Baas D., Lehmann S., Raposo G., Darlix J.L., Leblanc P. (2008). Mouse neuroblastoma cells release prion infectivity associated with exosomal vesiclesBiol Cell 100603618 [View Article][PubMed]. [Google Scholar]
  3. Barret A., Tagliavini F., Forloni G., Bate C., Salmona M., Colombo L., DeLuigi A., Limido L., Suardi S., other authors. (2003). Evaluation of quinacrine treatment for prion diseasesJ Virol 7784628469 [View Article][PubMed]. [Google Scholar]
  4. Brown P., Cervenáková L., Diringer H. (2001). Blood infectivity and the prospects for a diagnostic screening test in Creutzfeldt–Jakob diseaseJ Lab Clin Med 137513 [View Article][PubMed]. [Google Scholar]
  5. Caby M.-P., Lankar D., Vincendeau-Scherrer C., Raposo G., Bonnerot C. (2005). Exosomal-like vesicles are present in human blood plasmaInt Immunol 17879887 [View Article][PubMed]. [Google Scholar]
  6. Castilla J., Saá P., Soto C. (2005). Detection of prions in bloodNat Med 11982985[PubMed]. [Google Scholar]
  7. Coleman B.M., Hanssen E., Lawson V.A., Hill A.F. (2012). Prion-infected cells regulate the release of exosomes with distinct ultrastructural featuresFASEB J 2641604173 [View Article][PubMed]. [Google Scholar]
  8. Dickinson A.G. (1976). Scrapie in sheep and goats. In In Slow Virus Diseases of Animals and Man. Frontiers of Biology Series 44, pp. 209241. Edited by Kimberlin R. H.Amsterdam/New YorkNorth Holland. [Google Scholar]
  9. Douet J.Y., Zafar S., Perret-Liaudet A., Lacroux C., Lugan S., Aron N., Cassard H., Ponto C., Corbière F., other authors. (2014). Detection of infectivity in blood of persons with variant and sporadic Creutzfeldt–Jakob diseaseEmerg Infect Dis 20114117 [View Article][PubMed]. [Google Scholar]
  10. Edgeworth J.A., Farmer M., Sicilia A., Tavares P., Beck J., Campbell T., Lowe J., Mead S., Rudge P., other authors. (2011). Detection of prion infection in variant Creutzfeldt–Jakob disease: a blood-based assayLancet 377487493 [View Article][PubMed]. [Google Scholar]
  11. Fais S., Logozzi M., Lugini L., Federici C., Azzarito T., Zarovni N., Chiesi A. (2013). Exosomes: the ideal nanovectors for biodeliveryBiol Chem 394115 [View Article][PubMed]. [Google Scholar]
  12. Fauré J., Lachenal G., Court M., Hirrlinger J., Chatellard-Causse C., Blot B., Grange J., Schoehn G., Goldberg Y., other authors. (2006). Exosomes are released by cultured cortical neuronesMol Cell Neurosci 31642648 [View Article][PubMed]. [Google Scholar]
  13. Fevrier B., Vilette D., Archer F., Loew D., Faigle W., Vidal M., Laude H., Raposo G. (2004). Cells release prions in association with exosomesProc Natl Acad Sci U S A 10196839688 [View Article][PubMed]. [Google Scholar]
  14. Gregori L., Gurgel P.V., Lathrop J.T., Edwardson P., Lambert B.C., Carbonell R.G., Burton S.J., Hammond D.J., Rohwer R.G. (2006). Reduction in infectivity of endogenous transmissible spongiform encephalopathies present in blood by adsorption to selective affinity resinsLancet 36822262230 [View Article][PubMed]. [Google Scholar]
  15. Hartwell R.C., Nelson M.S., Kislan M.M., Stenland C.J., Miller J.L.C., Pifat D.Y., Petteway S.R. Jr, Cai K. (2005). An improved Western blot assay to assess the clearance of prion protein from plasma-derived therapeutic proteinsJ Virol Methods 125187193 [View Article][PubMed]. [Google Scholar]
  16. Houston F., McCutcheon S., Goldmann W., Chong A., Foster J., Sisó S., González L., Jeffrey M., Hunter N. (2008). Prion diseases are efficiently transmitted by blood transfusion in sheepBlood 11247394745 [View Article][PubMed]. [Google Scholar]
  17. Kimberlin R.H., Walker C. (1977). Characteristics of a short incubation model of scrapie in the golden hamsterJ Gen Virol 34295304 [View Article][PubMed]. [Google Scholar]
  18. Klöhn P.-C., Stoltze L., Flechsig E., Enari M., Weissmann C. (2003). A quantitative, highly sensitive cell-based infectivity assay for mouse scrapie prionsProc Natl Acad Sci U S A 1001166611671 [View Article][PubMed]. [Google Scholar]
  19. Lee D.C., Stenland C.J., Hartwell R.C., Ford E.K., Cai K., Miller J.L., Gilligan K.J., Rubenstein R., Fournel M., Petteway S.R. Jr (2000). Monitoring plasma processing steps with a sensitive Western blot assay for the detection of the prion proteinJ Virol Methods 847789 [View Article][PubMed]. [Google Scholar]
  20. Llewelyn C.A., Hewitt P.E., Knight R.S.G., Amar K., Cousens S., Mackenzie J., Will R.G. (2004). Possible transmission of variant Creutzfeldt–Jakob disease by blood transfusionLancet 363417421 [View Article][PubMed]. [Google Scholar]
  21. Meade-White K.D., Barbian K.D., Race B., Favara C., Gardner D., Taubner L., Porcella S., Race R. (2009). Characteristics of 263K scrapie agent in multiple hamster speciesEmerg Infect Dis 15207215 [View Article][PubMed]. [Google Scholar]
  22. Orrú C.D., Wilham J.M., Raymond L.D., Kuhn F., Schroeder B., Raeber A.J., Caughey B. (2011). Prion disease blood test using immunoprecipitation and improved quaking-induced conversionMBio 2e00078e00011 [View Article][PubMed]. [Google Scholar]
  23. Pan T., Sethi J., Nelsen C., Rudolph A., Cervenakova L., Brown P., Orser C.S. (2007). Detection of misfolded prion protein in blood with conformationally sensitive peptidesTransfusion 4714181425 [View Article][PubMed]. [Google Scholar]
  24. Puopolo M., Ladogana A., Vetrugno V., Pocchiari M. (2011). Transmission of sporadic Creutzfeldt–Jakob disease by blood transfusion: risk factor or possible biasesTransfusion 5115561566 [View Article][PubMed]. [Google Scholar]
  25. Raposo G., Nijman H.W., Stoorvogel W., Liejendekker R., Harding C.V., Melief C.J., Geuze H.J. (1996). B lymphocytes secrete antigen-presenting vesiclesJ Exp Med 1831161–1172 [View Article][PubMed]. [Google Scholar]
  26. Saá P., Castilla J., Soto C. (2006). Presymptomatic detection of prions in bloodScience 3139294 [View Article][PubMed]. [Google Scholar]
  27. Saá P., Yakovleva O., De Castro J., Vasilyeva I., De Paoli S.H., Simak J., Cervenakova L. (2014). First demonstration of transmissible spongiform encephalopathy-associated prion protein (PrPTSE) in extracellular vesicles from plasma of mice infected with mouse-adapted variant Creutzfeldt–Jakob disease by in vitro amplificationJ Biol Chem 2892924729260 [View Article][PubMed]. [Google Scholar]
  28. Théry C., Ostrowski M., Segura E. (2009). Membrane vesicles as conveyors of immune responsesNat Rev Immunol 9581593 [View Article][PubMed]. [Google Scholar]
  29. Vella L.J., Sharples R.A., Lawson V.A., Masters C.L., Cappai R., Hill A.F. (2007). Packaging of prions into exosomes is associated with a novel pathway of PrP processingJ Pathol 211582590 [View Article][PubMed]. [Google Scholar]
  30. Weissmann C. (2004). The state of the prionNat Rev Microbiol 2861871 [View Article][PubMed]. [Google Scholar]
  31. Wroe S.J., Pal S., Siddique D., Hyare H., Macfarlane R., Joiner S., Linehan J.M., Brandner S., Wadsworth J.D.F., other authors. (2006). Clinical presentation and pre-mortem diagnosis of variant Creutzfeldt–Jakob disease associated with blood transfusion: a case reportLancet 36820612067 [View Article][PubMed]. [Google Scholar]

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