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Volume 23, Number 6—June 2017

Distribution and Quantitative Estimates of Variant Creutzfeldt-Jakob Disease Prions in Tissues of Clinical and Asymptomatic Patients

Jean Y. Douet, Caroline Lacroux, Naima Aron, Mark W. Head, Séverine Lugan, Cécile Tillier, Alvina Huor, Hervé Cassard, Mark Arnold, Vincent Beringue, James W. Ironside, and Olivier AndréolettiComments to Author 
Author affiliations: Institut National de la Recherche Agronomique, Toulouse, France (J.Y. Douet, C. Lacroux, N. Aron, S. Lugan, C. Tillier, A. Huor, H. Cassard, O. Andréoletti); University of Edinburgh, Edinburgh, Scotland, UK (M.W. Head, J.W. Ironside); Animal and Plant Health Agency, Loughborough, UK (M. Arnold); Institut National de la Recherche Agronomique, Jouy-en-Josas, France (V. Beringue)

Main Article

Table 5

Bone marrow sample bioassay in bovine PrP–expressing mice (tgBov) for 4 patients with vCJD*

Sample Transmission in tgBov mice
Mean infectious titer, LD50/g (95% CI)
No. positive/no. inoculated mice Mean ± SD incubation, d
vCJD-1 5/5 458 ± 37 103.1 (102.6–103.5)
vCJD-2 6/6 373 ± 35 104.7 (104.3–05.2)
vCJD-3 4/6 504 ± 10 102.3 (101.8–102.7)
vCJD-4 6/6 447 ± 91 104.0 (103.4–04.5)
PBS control 0/6 >600 NA

*A 10% wt/vol bone marrow homogenate prepared from 4 symptomatic vCJD patients (Table 3) was inoculated intracerebrally into 6 tgBov mice (20 µL/mouse). One mouse (inoculated with homogenate from patient vCJD-1) died within the first few days after intercerebral inoculation. Mice were euthanized when they showed clinical signs of prion infection or after 600-d postinoculation. Mice were considered prion infected when abnormal PrP deposition was detected in brain. Infectious prion titers were estimated by using the method of Arnold et al. (17). The method uses the probability of survival (attack rate at each dilution) and the individual mouse incubation periods at each dilution to estimate infectious load. Infectious titers are given as estimated values. LD50, 50% lethal dose; NA, not applicable; PBS, phosphate-buffered saline; PrP; prion protein; vCJD, variant Creutzfeldt-Jakob disease.

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  1. McKinley  MP, Bolton  DC, Prusiner  SB. A protease-resistant protein is a structural component of the scrapie prion. Cell. 1983;35:5762. DOIPubMedGoogle Scholar
  2. Bruce  ME, Will  RG, Ironside  JW, McConnell  I, Drummond  D, Suttie  A, et al. Transmissions to mice indicate that ‘new variant’ CJD is caused by the BSE agent. Nature. 1997;389:498501. DOIPubMedGoogle Scholar
  3. Collinge  J, Sidle  KC, Meads  J, Ironside  J, Hill  AF. Molecular analysis of prion strain variation and the aetiology of ‘new variant’ CJD. Nature. 1996;383:68590. DOIPubMedGoogle Scholar
  4. Garske  T, Ghani  AC. Uncertainty in the tail of the variant Creutzfeldt-Jakob disease epidemic in the UK. PLoS One. 2010;5:e15626. DOIPubMedGoogle Scholar
  5. Gill  ON, Spencer  Y, Richard-Loendt  A, Kelly  C, Dabaghian  R, Boyes  L, et al. Prevalent abnormal prion protein in human appendixes after bovine spongiform encephalopathy epizootic: large scale survey. BMJ. 2013;347(oct15 5):f5675. DOIPubMedGoogle Scholar
  6. Wadsworth  JD, Joiner  S, Hill  AF, Campbell  TA, Desbruslais  M, Luthert  PJ, et al. Tissue distribution of protease resistant prion protein in variant Creutzfeldt-Jakob disease using a highly sensitive immunoblotting assay. Lancet. 2001;358:17180. DOIPubMedGoogle Scholar
  7. Haïk  S, Faucheux  BA, Sazdovitch  V, Privat  N, Kemeny  JL, Perret-Liaudet  A, et al. The sympathetic nervous system is involved in variant Creutzfeldt-Jakob disease. Nat Med. 2003;9:11213. DOIPubMedGoogle Scholar
  8. Head  MW, Ritchie  D, Smith  N, McLoughlin  V, Nailon  W, Samad  S, et al. Peripheral tissue involvement in sporadic, iatrogenic, and variant Creutzfeldt-Jakob disease: an immunohistochemical, quantitative, and biochemical study. Am J Pathol. 2004;164:14353. DOIPubMedGoogle Scholar
  9. Saborio  GP, Permanne  B, Soto  C. Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding. Nature. 2001;411:8103. DOIPubMedGoogle Scholar
  10. Moda  F, Gambetti  P, Notari  S, Concha-Marambio  L, Catania  M, Park  KW, et al. Prions in the urine of patients with variant Creutzfeldt-Jakob disease. N Engl J Med. 2014;371:5309. DOIPubMedGoogle Scholar
  11. Lacroux  C, Comoy  E, Moudjou  M, Perret-Liaudet  A, Lugan  S, Litaise  C, et al. Preclinical detection of variant CJD and BSE prions in blood. PLoS Pathog. 2014;10:e1004202. DOIPubMedGoogle Scholar
  12. Peden  AH, Head  MW, Ritchie  DL, Bell  JE, Ironside  JW. Preclinical vCJD after blood transfusion in a PRNP codon 129 heterozygous patient. Lancet. 2004;364:5279. DOIPubMedGoogle Scholar
  13. Uro-Coste  E, Cassard  H, Simon  S, Lugan  S, Bilheude  JM, Perret-Liaudet  A, et al. Beyond PrPres type 1/type 2 dichotomy in Creutzfeldt-Jakob disease. PLoS Pathog. 2008;4:e1000029. DOIGoogle Scholar
  14. Moreno  CR, Moazami-Goudarzi  K, Laurent  P, Cazeau  G, Andreoletti  O, Chadi  S, et al. Which PrP haplotypes in a French sheep population are the most susceptible to atypical scrapie? Arch Virol. 2007;152:122932. DOIPubMedGoogle Scholar
  15. Castilla  J, Gutiérrez Adán  A, Brun  A, Pintado  B, Ramírez  MA, Parra  B, et al. Early detection of PrPres in BSE-infected bovine PrP transgenic mice. Arch Virol. 2003;148:67791. DOIPubMedGoogle Scholar
  16. Douet  JY, Zafar  S, Perret-Liaudet  A, Lacroux  C, Lugan  S, Aron  N, et al. Detection of infectivity in blood of persons with variant and sporadic Creutzfeldt-Jakob disease. Emerg Infect Dis. 2014;20:1147. DOIPubMedGoogle Scholar
  17. Arnold  ME, Hawkins  SA, Green  R, Dexter  I, Wells  GA. Pathogenesis of experimental bovine spongiform encephalopathy (BSE): estimation of tissue infectivity according to incubation period. Vet Res. 2009;40:8. DOIPubMedGoogle Scholar
  18. Groschup  MH, Buschmann  A. Rodent models for prion diseases. Vet Res. 2008;39:32. DOIPubMedGoogle Scholar
  19. Douet  JY, Lacroux  C, Corbière  F, Litaise  C, Simmons  H, Lugan  S, et al. PrP expression level and sensitivity to prion infection. J Virol. 2014;88:58702. DOIPubMedGoogle Scholar
  20. Féraudet  C, Morel  N, Simon  S, Volland  H, Frobert  Y, Créminon  C, et al. Screening of 145 anti-PrP monoclonal antibodies for their capacity to inhibit PrPSc replication in infected cells. J Biol Chem. 2005;280:1124758. DOIPubMedGoogle Scholar
  21. Langeveld  JP, Jacobs  JG, Erkens  JH, Bossers  A, van Zijderveld  FG, van Keulen  LJ. Rapid and discriminatory diagnosis of scrapie and BSE in retro-pharyngeal lymph nodes of sheep. BMC Vet Res. 2006;2:19. DOIPubMedGoogle Scholar
  22. Cassard  H, Torres  JM, Lacroux  C, Douet  JY, Benestad  SL, Lantier  F, et al. Evidence for zoonotic potential of ovine scrapie prions. Nat Commun. 2014;5:5821. DOIPubMedGoogle Scholar
  23. Lacroux  C, Corbière  F, Tabouret  G, Lugan  S, Costes  P, Mathey  J, et al. Dynamics and genetics of PrPSc placental accumulation in sheep. J Gen Virol. 2007;88:105661. DOIPubMedGoogle Scholar
  24. Andréoletti  O, Berthon  P, Marc  D, Sarradin  P, Grosclaude  J, van Keulen  L, et al. 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. 2000;81:311526. DOIPubMedGoogle Scholar
  25. Foster  JD, Parnham  DW, Hunter  N, Bruce  M. Distribution of the prion protein in sheep terminally affected with BSE following experimental oral transmission. J Gen Virol. 2001;82:231926. DOIPubMedGoogle Scholar
  26. Lacroux  C, Vilette  D, Fernández-Borges  N, Litaise  C, Lugan  S, Morel  N, et al. Prionemia and leukocyte-platelet-associated infectivity in sheep transmissible spongiform encephalopathy models. J Virol. 2012;86:205666. DOIPubMedGoogle Scholar
  27. Houston  F, Foster  JD, Chong  A, Hunter  N, Bostock  CJ. Transmission of BSE by blood transfusion in sheep. Lancet. 2000;356:9991000. DOIPubMedGoogle Scholar
  28. Hilton  DA. Pathogenesis and prevalence of variant Creutzfeldt-Jakob disease. J Pathol. 2006;208:13441. DOIPubMedGoogle Scholar
  29. Tamgüney  G, Richt  JA, Hamir  AN, Greenlee  JJ, Miller  MW, Wolfe  LL, et al. Salivary prions in sheep and deer. Prion. 2012;6:5261. DOIPubMedGoogle Scholar
  30. Andréoletti  O, Simon  S, Lacroux  C, Morel  N, Tabouret  G, Chabert  A, et al. PrPSc accumulation in myocytes from sheep incubating natural scrapie. Nat Med. 2004;10:5913. DOIPubMedGoogle Scholar
  31. Chianini  F, Cosseddu  GM, Steele  P, Hamilton  S, Hawthorn  J, Síso  S, et al. Correlation between infectivity and disease associated prion protein in the nervous system and selected edible tissues of naturally affected scrapie sheep. PLoS One. 2015;10:e0122785. DOIPubMedGoogle Scholar
  32. Hadlow  WJ, Kennedy  RC, Race  RE. Natural infection of Suffolk sheep with scrapie virus. J Infect Dis. 1982;146:65764. DOIPubMedGoogle Scholar
  33. Garza  MC, Monzón  M, Marín  B, Badiola  JJ, Monleón  E. Distribution of peripheral PrP(Sc) in sheep with naturally acquired scrapie. PLoS One. 2014;9:e97768. DOIPubMedGoogle Scholar
  34. Castilla  J, Saá  P, Soto  C. Detection of prions in blood. Nat Med. 2005;11:9825.PubMedGoogle Scholar
  35. Bougard  D, Brandel  JP, Bélondrade  M, Béringue  V, Segarra  C, Fleury  H, et al. Detection of prions in the plasma of presymptomatic and symptomatic patients with variant Creutzfeldt-Jakob disease. Sci Transl Med. 2016;8:370ra182. DOIPubMedGoogle Scholar
  36. Concha-Marambio  L, Pritzkow  S, Moda  F, Tagliavini  F, Ironside  JW, Schulz  PE, et al. Detection of prions in blood from patients with variant Creutzfeldt-Jakob disease. Sci Transl Med. 2016;8:370ra183. DOIPubMedGoogle Scholar
  37. Bishop  MT, Diack  AB, Ritchie  DL, Ironside  JW, Will  RG, Manson  JC. Prion infectivity in the spleen of a PRNP heterozygous individual with subclinical variant Creutzfeldt-Jakob disease. Brain. 2013;136:113945. DOIPubMedGoogle Scholar
  38. Sisó  S, González  L, Jeffrey  M. Neuroinvasion in prion diseases: the roles of ascending neural infection and blood dissemination. Interdiscip Perspect Infect Dis. 2010;2010:747892.
  39. Brown  P, Brandel  JP, Sato  T, Nakamura  Y, MacKenzie  J, Will  RG, et al. Iatrogenic Creutzfeldt-Jakob disease, final assessment. Emerg Infect Dis. 2012;18:9017. DOIPubMedGoogle Scholar
  40. Flechsig  E, Hegyi  I, Enari  M, Schwarz  P, Collinge  J, Weissmann  C. Transmission of scrapie by steel-surface-bound prions. Mol Med. 2001;7:67984.PubMedGoogle Scholar
  41. Taylor  D. Inactivation of the BSE agent. C R Biol. 2002;325:756. DOIPubMedGoogle Scholar
  42. Molesworth  A, Yates  P, Hewitt  PE, Mackenzie  J, Ironside  JW, Galea  G, et al. Investigation of variant Creutzfeldt-Jakob disease implicated organ or tissue transplantation in the United Kingdom. Transplantation. 2014;98:5859. DOIPubMedGoogle Scholar
  43. Jayanthi  P, Thomas  P, Bindhu  P, Krishnapillai  R. Prion diseases in humans: oral and dental implications. N Am J Med Sci. 2013;5:399403. DOIPubMedGoogle Scholar
  44. Peden  A, McCardle  L, Head  MW, Love  S, Ward  HJ, Cousens  SN, et al. Variant CJD infection in the spleen of a neurologically asymptomatic UK adult patient with haemophilia. Haemophilia. 2010;16:296304. DOIPubMedGoogle Scholar
  45. Llewelyn  CA, Hewitt  PE, Knight  RS, Amar  K, Cousens  S, Mackenzie  J, et al. Possible transmission of variant Creutzfeldt-Jakob disease by blood transfusion. Lancet. 2004;363:41721. DOIPubMedGoogle Scholar
  46. Lefrère  JJ, Hewitt  P. From mad cows to sensible blood transfusion: the risk of prion transmission by labile blood components in the United Kingdom and in France. Transfusion. 2009;49:797812. DOIPubMedGoogle Scholar
  47. Edgeworth  JA, Farmer  M, Sicilia  A, Tavares  P, Beck  J, Campbell  T, et al. Detection of prion infection in variant Creutzfeldt-Jakob disease: a blood-based assay. Lancet. 2011;377:48793. DOIPubMedGoogle Scholar
  48. Jackson  GS, Burk-Rafel  J, Edgeworth  JA, Sicilia  A, Abdilahi  S, Korteweg  J, et al. A highly specific blood test for vCJD. Blood. 2014;123:4523. DOIPubMedGoogle Scholar

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