Prions exist as different strains exhibiting distinct disease phenotypes. Currently, the identification of prion strains is still based on biological strain typing in rodents. However, it has been shown that prion strains may be associated with distinct PrP Sc biochemical types. Taking advantage of the availability of several prion strains adapted to a novel rodent model, the bank vole, we investigated if any prion strain was actually associated with distinctive PrP Sc biochemical characteristics and if it was possible to univocally identify strains through PrP Sc biochemical phenotypes. We selected six different vole-adapted strains (three human-derived and three animal-derived) and analyzed PrP Sc from individual voles by epitope mapping of protease resistant core of PrP Sc (PrP res) and by conformational stability and solubility assay. Overall, we discriminated five out of six prion strains, while two different scrapie strains showed identical PrP Sc types. Our results suggest that the biochemical strain typing approach here proposed was highly discriminative, although by itself it did not allow us to identify all prion strains analyzed.
Bruce, M.E.; Fraser, H. Scrapie strain variation and its implications. Curr. Topics Microbiol. Immunol. 1991, 172, 125–138, doi:10.1007/978-3-642-76540-7_8.
[3]
Beringue, V.; Vilotte, J.L.; Laude, H. Prion agent diversity and species barrier. Vet. Res. 2008, 39, doi:10.1051/vetres:2008024.
[4]
Telling, G.C.; Parchi, P.; DeArmond, S.J.; Cortelli, P.; Montagna, P.; Gabizon, R.; Mastrianni, J.; Lugaresi, E.; Gambetti, P.; Prusiner, S.B. Evidence for the conformation of the pathologic isoform of the prion protein enciphering and propagating prion diversity. Science 1996, 274, 2079–2082, doi:10.1126/science.274.5295.2079.
[5]
Collinge, J.; Sidle, K.C.; Meads, J.; Ironside, J.; Hill, A.F. Molecular analysis of prion strain variation and the aetiology of 'new variant' cjd. Nature 1996, 383, 685–690, doi:10.1038/383685a0.
[6]
Parchi, P.; Castellani, R.; Capellari, S.; Ghetti, B.; Young, K.; Chen, S.G.; Farlow, M.; Dickson, D.W.; Sima, A.A.; Trojanowski, J.Q.; et al. Molecular basis of phenotypic variability in sporadic creutzfeldt-jakob disease. Ann. Neurol. 1996, 39, 767–778, doi:10.1002/ana.410390613.
[7]
Bessen, R.A.; Marsh, R.F. Distinct prp properties suggest the molecular basis of strain variation in transmissible mink encephalopathy. J. Virol. 1994, 68, 7859–7868.
[8]
Kascsak, R.J.; Rubenstein, R.; Merz, P.A.; Carp, R.I.; Wisniewski, H.M.; Diringer, H. Biochemical differences among scrapie-associated fibrils support the biological diversity of scrapie agents. J. Gen. Virol. 1985, 66, 1715–1722, doi:10.1099/0022-1317-66-8-1715.
[9]
Kuczius, T.; Groschup, M.H. Differences in proteinase k resistance and neuronal deposition of abnormal prion proteins characterize bovine spongiform encephalopathy (bse) and scrapie strains. Mol. Med. 1999, 5, 406–418.
[10]
Peretz, D.; Scott, M.R.; Groth, D.; Williamson, R.A.; Burton, D.R.; Cohen, F.E.; Prusiner, S.B. Strain-specified relative conformational stability of the scrapie prion protein. Protein Sci. 2001, 10, 854–863, doi:10.1110/ps.39201.
[11]
Pirisinu, L.; Di Bari, M.; Marcon, S.; Vaccari, G.; D’Agostino, C.; Fazzi, P.; Esposito, E.; Galeno, R.; Langeveld, J.; Agrimi, U.; et al. A new method for the characterization of strain-specific conformational stability of protease-sensitive and protease-resistant prp. PloS One 2010, 5, e12723, doi:10.1371/journal.pone.0012723.
[12]
Safar, J.; Wille, H.; Itri, V.; Groth, D.; Serban, H.; Torchia, M.; Cohen, F.E.; Prusiner, S.B. Eight prion strains have prp(sc) molecules with different conformations. Nat. Med. 1998, 4, 1157–1165, doi:10.1038/2654.
[13]
Stack, M.J.; Chaplin, M.J.; Clark, J. Differentiation of prion protein glycoforms from naturally occurring sheep scrapie, sheep-passaged scrapie strains (ch1641 and ssbp1), bovine spongiform encephalopathy (bse) cases and romney and cheviot breed sheep experimentally inoculated with bse using two monoclonal antibodies. Acta Neuropathol. 2002, 104, 279–286, doi:10.1007/s00401-002-0556-2.
[14]
Stack, M.; Jeffrey, M.; Gubbins, S.; Grimmer, S.; Gonzalez, L.; Martin, S.; Chaplin, M.; Webb, P.; Simmons, M.; Spencer, Y.; et al. Monitoring for bovine spongiform encephalopathy in sheep in great britain, 1998–2004. J. Gen. Virol. 2006, 87, 2099–2107, doi:10.1099/vir.0.81254-0.
[15]
Vulin, J.; Biacabe, A.G.; Cazeau, G.; Calavas, D.; Baron, T. Molecular typing of protease-resistant prion protein in transmissible spongiform encephalopathies of small ruminants, france, 2002–2009. Emerg. Infect. Dis. 2011, 17, 55–63, doi:10.3201/eid1701.100891.
[16]
Hope, J.; Wood, S.C.; Birkett, C.R.; Chong, A.; Bruce, M.E.; Cairns, D.; Goldmann, W.; Hunter, N.; Bostock, C.J. Molecular analysis of ovine prion protein identifies similarities between bse and an experimental isolate of natural scrapie, ch1641. J. Gen. Virol. 1999, 80, 1–4.
[17]
Hill, A.F.; Sidle, K.C.; Joiner, S.; Keyes, P.; Martin, T.C.; Dawson, M.; Collinge, J. Molecular screening of sheep for bovine spongiform encephalopathy. Neurosci. Lett. 1998, 255, 159–162, doi:10.1016/S0304-3940(98)00736-8.
[18]
Saa, P.; Sferrazza, G.F.; Ottenberg, G.; Oelschlegel, A.M.; Dorsey, K.; Lasmezas, C.I. Strain-specific role of rnas in prion replication. J. Virol. 2012, 86, 10494–10504, doi:10.1128/JVI.01286-12.
[19]
Tzaban, S.; Friedlander, G.; Schonberger, O.; Horonchik, L.; Yedidia, Y.; Shaked, G.; Gabizon, R.; Taraboulos, A. Protease-sensitive scrapie prion protein in aggregates of heterogeneous sizes. Biochemistry 2002, 41, 12868–12875, doi:10.1021/bi025958g.
[20]
Pastrana, M.A.; Sajnani, G.; Onisko, B.; Castilla, J.; Morales, R.; Soto, C.; Requena, J.R. Isolation and characterization of a proteinase k-sensitive prpsc fraction. Biochemistry 2006, 45, 15710–15717, doi:10.1021/bi0615442.
[21]
Thackray, A.M.; Hopkins, L.; Bujdoso, R. Proteinase k-sensitive disease-associated ovine prion protein revealed by conformation-dependent immunoassay. Biochem. J. 2007, 401, 475–483, doi:10.1042/BJ20061264.
[22]
Cronier, S.; Gros, N.; Tattum, M.H.; Jackson, G.S.; Clarke, A.R.; Collinge, J.; Wadsworth, J.D. Detection and characterization of proteinase k-sensitive disease-related prion protein with thermolysin. Biochem. J. 2008, 416, 297–305, doi:10.1042/BJ20081235.
[23]
Gambetti, P.; Dong, Z.; Yuan, J.; Xiao, X.; Zheng, M.; Alshekhlee, A.; Castellani, R.; Cohen, M.; Barria, M.A.; Gonzalez-Romero, D.; et al. A novel human disease with abnormal prion protein sensitive to protease. Ann. Neurol. 2008, 63, 697–708.
[24]
Nonno, R.; Di Bari, M.A.; Cardone, F.; Vaccari, G.; Fazzi, P.; Dell’Omo, G.; Cartoni, C.; Ingrosso, L.; Boyle, A.; Galeno, R.; et al. Efficient transmission and characterization of creutzfeldt-jakob disease strains in bank voles. PLoS Pathog. 2006, 2, e12, doi:10.1371/journal.ppat.0020012.
[25]
Zanusso, G.; Polo, A.; Farinazzo, A.; Nonno, R.; Cardone, F.; Di Bari, M.; Ferrari, S.; Principe, S.; Gelati, M.; Fasoli, E.; et al. Novel prion protein conformation and glycotype in creutzfeldt-jakob disease. Arch. Neurol. 2007, 64, 595–599, doi:10.1001/archneur.64.4.595.
[26]
Di Bari, M.A.; Chianini, F.; Vaccari, G.; Esposito, E.; Conte, M.; Eaton, S.L.; Hamilton, S.; Finlayson, J.; Steele, P.J.; Dagleish, M.P.; et al. The bank vole (myodes glareolus) as a sensitive bioassay for sheep scrapie. J. Gen. Virol. 2008, 89, 2975–2985, doi:10.1099/vir.0.2008/005520-0.
[27]
Piening, N.; Nonno, R.; Di Bari, M.; Walter, S.; Windl, O.; Agrimi, U.; Kretzschmar, H.A.; Bertsch, U. Conversion efficiency of bank vole prion protein in vitro is determined by residues 155 and 170, but does not correlate with the high susceptibility of bank voles to sheep scrapie in vivo. J. Biol. Chem. 2006, 281, 9373–9384.
[28]
Agrimi, U.; Nonno, R.; Dell’Omo, G.; Di Bari, M.A.; Conte, M.; Chiappini, B.; Esposito, E.; Di Guardo, G.; Windl, O.; Vaccari, G.; et al. Prion protein amino acid determinants of differential susceptibility and molecular feature of prion strains in mice and voles. PLoS Pathog. 2008, 4, e1000113, doi:10.1371/journal.ppat.1000113.
[29]
Di Bari, M.A.; Nonno, R.; Castilla, J.; D'Agostino, C.; Pirisinu, L.; Riccardi, G.; Conte, M.; Richt, J.; Kunkle, R.; Langeveld, J.; Vaccari, G.; Agrimi, U. Chronic wasting disease in bank voles: Characterisation of the shortest incubation time model for prion diseases. PLoS Pathog. 2013, 9, e1003219, doi:10.1371/journal.ppat.1003219.
[30]
Cartoni, C.; Schinina, M.E.; Maras, B.; Nonno, R.; Vaccari, G.; Di Bari, M.; Conte, M.; de Pascalis, A.; Principe, S.; Cardone, F.; et al. Quantitative profiling of the pathological prion protein allotypes in bank voles by liquid chromatography-mass spectrometry. J. Chromatogr. B 2007, 849, 302–306, doi:10.1016/j.jchromb.2006.08.016.
[31]
Bruce, M.E.; Boyle, A.; Cousens, S.; McConnell, I.; Foster, J.; Goldmann, W.; Fraser, H. Strain characterization of natural sheep scrapie and comparison with bse. J. Gen. Virol. 2002, 83, 695–704.
[32]
Notari, S.; Strammiello, R.; Capellari, S.; Giese, A.; Cescatti, M.; Grassi, J.; Ghetti, B.; Langeveld, J.P.; Zou, W.Q.; Gambetti, P.; et al. Characterization of truncated forms of abnormal prion protein in creutzfeldt-jakob disease. J. Biol. Chem. 2008, 283, 30557–30565, doi:10.1074/jbc.M801877200.
[33]
Zou, W.Q.; Capellari, S.; Parchi, P.; Sy, M.S.; Gambetti, P.; Chen, S.G. Identification of novel proteinase k-resistant c-terminal fragments of prp in creutzfeldt-jakob disease. J. Biol. Chem. 2003, 278, 40429–40436.
[34]
Notari, S.; Capellari, S.; Giese, A.; Westner, I.; Baruzzi, A.; Ghetti, B.; Gambetti, P.; Kretzschmar, H.A.; Parchi, P. Effects of different experimental conditions on the prpsc core generated by protease digestion: Implications for strain typing and molecular classification of cjd. J. Biol. Chem. 2004, 279, 16797–16804, doi:10.1074/jbc.M313220200.
[35]
Pirisinu, L.; Migliore, S.; Di Bari, M.A.; Esposito, E.; Baron, T.; D’Agostino, C.; De Grossi, L.; Vaccari, G.; Agrimi, U.; Nonno, R. Molecular discrimination of sheep bovine spongiform encephalopathy from scrapie. Emerg. Infect. Dis. 2011, 17, 695–698, doi:10.3201/eid1704.101215.
[36]
Legname, G.; Nguyen, H.O.; Peretz, D.; Cohen, F.E.; DeArmond, S.J.; Prusiner, S.B. Continuum of prion protein structures enciphers a multitude of prion isolate-specified phenotypes. Proc. Natl. Acad. Sci. USA 2006, 103, 19105–19110.
[37]
Ayers, J.I.; Schutt, C.R.; Shikiya, R.A.; Aguzzi, A.; Kincaid, A.E.; Bartz, J.C. The strain-encoded relationship between prp replication, stability and processing in neurons is predictive of the incubation period of disease. PLoS Pathog. 2011, 7, e1001317, doi:10.1371/journal.ppat.1001317.
[38]
Bett, C.; Joshi-Barr, S.; Lucero, M.; Trejo, M.; Liberski, P.; Kelly, J.W.; Masliah, E.; Sigurdson, C.J. Biochemical properties of highly neuroinvasive prion strains. PLoS Pathog. 2012, 8, e1002522, doi:10.1371/journal.ppat.1002522.
[39]
Pirisinu, L.; Nonno, R.; Esposito, E.; Benestad, S.L.; Gambetti, P.; Agrimi, U.; Zou, W.Q. Small ruminant Nor98 prions share biochemical features with human Gerstmann-Str？ussler-Scheinker disease and variably protease-sensitive prionopathy. PLoS One 2013, 8, e66405, doi:10.1371/journal.pone.0066405.
