Detection of exosomal prions in blood by immunochemistry techniques.

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, PrPTSE) in blood remains elusive. We developed a novel method for the detection of PrPTSE in blood of prion-infected rodents based on the finding that PrPTSE is associated with plasma exosomes. However, further purification of the exosomes on a sucrose gradient was necessary to remove plasma immunoglobulins, which interfere with PrPTSE, masking its detection by immunochemistry. Finally, we report that about 20% of plasma infectivity is associated with exosomes.

[1]  H. Abdel-Haq.  Factors intrinsic and extrinsic to blood hamper the development of a routine blood test for human prion diseases. , 2015, The Journal of general virology.

[2]  P. Saá,et al.  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 Amplification* , 2014, The Journal of Biological Chemistry.

[3]  I. Zerr,et al.  Detection of Infectivity in Blood of Persons with Variant and Sporadic Creutzfeldt-Jakob Disease , 2014, Emerging infectious diseases.

[4]  A. Hill,et al.  Prion‐infected cells regulate the release of exosomes with distinct ultrastructural features , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  M. Puopolo,et al.  Transmission of sporadic Creutzfeldt‐Jakob disease by blood transfusion: risk factor or possible biases , 2011, Transfusion.

[6]  B. Caughey,et al.  Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion , 2011, mBio.

[7]  J. Collinge,et al.  Detection of prion infection in variant Creutzfeldt-Jakob disease: a blood-based assay , 2011, The Lancet.

[8]  C. Théry,et al.  Membrane vesicles as conveyors of immune responses , 2009, Nature Reviews Immunology.

[9]  C. Favara,et al.  Characteristics of 263K Scrapie Agent in Multiple Hamster Species , 2009, Emerging infectious diseases.

[10]  N. Hunter,et al.  Prion diseases are efficiently transmitted by blood transfusion in sheep. , 2008, Blood.

[11]  G. Raposo,et al.  Mouse neuroblastoma cells release prion infectivity associated with exosomal vesicles , 2008, Biology of the cell.

[12]  P. Brown,et al.  Detection of misfolded prion protein in blood with conformationally sensitive peptides , 2007, Transfusion.

[13]  R. Cappai,et al.  Packaging of prions into exosomes is associated with a novel pathway of PrP processing , 2007, The Journal of pathology.

[14]  R. Carbonell,et al.  Reduction in infectivity of endogenous transmissible spongiform encephalopathies present in blood by adsorption to selective affinity resins , 2006, The Lancet.

[15]  Suvankar Pal,et al.  Clinical presentation and pre-mortem diagnosis of variant Creutzfeldt-Jakob disease associated with blood transfusion: a case report , 2006, The Lancet.

[16]  J. Castilla,et al.  Presymptomatic Detection of Prions in Blood , 2006, Science.

[17]  G. Lachenal,et al.  Exosomes are released by cultured cortical neurones , 2006, Molecular and Cellular Neuroscience.

[18]  J. Castilla,et al.  Detection of prions in blood , 2005, Nature Medicine.

[19]  Graça Raposo,et al.  Exosomal-like vesicles are present in human blood plasma. , 2005, International immunology.

[20]  C. Stenland,et al.  An improved Western blot assay to assess the clearance of prion protein from plasma-derived therapeutic proteins. , 2005, Journal of virological methods.

[21]  Charles Weissmann,et al.  The state of the prion , 2004, Nature Reviews Microbiology.

[22]  W. Faigle,et al.  Cells release prions in association with exosomes. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[23]  J. MacKenzie,et al.  Possible transmission of variant Creutzfeldt-Jakob disease by blood transfusion , 2004, The Lancet.

[24]  L. Stoltze,et al.  A quantitative, highly sensitive cell-based infectivity assay for mouse scrapie prions , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[25]  G. Forloni,et al.  Evaluation of Quinacrine Treatment for Prion Diseases , 2003, Journal of Virology.

[26]  C. Melief,et al.  B lymphocytes secrete antigen-presenting vesicles , 1996, The Journal of experimental medicine.

[27]  R. Kimberlin,et al.  Characteristics of a short incubation model of scrapie in the golden hamster. , 1977, The Journal of general virology.

[28]  M. Logozzi,et al.  Exosomes: the ideal nanovectors for biodelivery , 2013, Biological chemistry.

[29]  P. Brown,et al.  Blood infectivity and the prospects for a diagnostic screening test in Creutzfeldt-Jakob disease. , 2001, The Journal of laboratory and clinical medicine.

[30]  J. Miller,et al.  Monitoring plasma processing steps with a sensitive Western blot assay for the detection of the prion protein. , 2000, Journal of virological methods.

[31]  Dickinson Ag.  Scrapie in sheep and goats. , 1976 .