A Rapid Bioassay for Classical and L-Type Bovine Spongiform Encephalopathies


The rapid detection of infectivity of several agents that cause Creutzfeldt-Jakob disease has previously been achieved by assaying for deposits of abnormal prion protein (PrPSc) in follicular dendritic cells in the spleens of transgenic mice carrying the human prion protein gene. In this study, transgenic mice expressing the bovine prion protein were inoculated intraperitoneally with classical (C-type) or atypical L-type bovine spongiform encephalopathies (BSE). Proteinase-resistant PrPSc were detected in the spleens of all transgenic mice at 75 days after inoculation with both types of BSE. Infectivity in PrPSc-positive spleens of the transgenic mice revealed that prions of C- and L-type BSE replicated. These results suggest that bioassay system by the transgenic mice could be useful for the rapid detection of BSE infectivity with discriminating between C- and L-type BSEs.

Share and Cite:

Y. Matsuura, Y. Ishikawa, R. Somerville, T. Yokoyama, K. Hagiwara, Y. Yamakawa, T. Sata, T. Kitamoto and S. Mohri, "A Rapid Bioassay for Classical and L-Type Bovine Spongiform Encephalopathies," Open Journal of Veterinary Medicine, Vol. 3 No. 1, 2013, pp. 79-85. doi: 10.4236/ojvm.2013.31013.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] G. A. Wells, A. C. Scott, C. T. Johnson, R. F. Gunning, R. D. Hancock, M. Jeffrey, M. Dawson and R. Bradley, “A Novel Progressive Spongiform Encephalopathy in Cattle,” Veterinary Record, Vol. 121, No. 18, 1987, pp. 419- 420. doi:10.1136/vr.121.18.419
[2] G. A. Wells and J. W. Wilesmith, “The Neuropathology and Epidemiology of Bovine Spongiform Encephalopathy,” Brain Pathology, Vol. 5, No. 1, 1995, pp. 91-103. doi:10.1111/j.1750-3639.1995.tb00580.x
[3] J. Collinge, K. C. Sidle, J. Meads, J. Ironside and A. F. Hill, “Molecular Analysis of Prion Strain Variation and the Aetiology of ‘New Variant’ CJD,” Nature, Vol. 383, No. 6602, 1996, pp. 685-690. doi:10.1038/383685a0
[4] A. F. Hill, M. Desbruslais, S. Joiner, K. C. Sidle, I. Gowland, J. Collinge, L. J. Doey and P. Lantos, “The Same Prion Strain Causes vCJD and BSE,” Nature, Vol. 389, No. 6650, 1997, pp. 448-450, 526. doi:10.1038/38925
[5] T. Kuczius, I. Haist and M. H. Groschup, “Molecular Analysis of Bovine Spongiform Encephalopathy and Scrapie Strain Variation,” Journal of Infectious Diseases, Vol. 178, No. 3, 1998, pp. 693-699. doi:10.1086/515337
[6] M. E. Bruce, R. G. Will, J. W. Ironside, I. McConnell, D. Drummond, A. Suttie, L. McCardle, A. Chree, J. Hope, C. Birkett, S. Cousens, H. Fraser and C. J. Bostock, “Transmissions to Mice Indicate That ‘New Variant’ CJD Is Caused by the BSE Agent,” Nature, Vol. 389, No. 6650, 1997, pp. 498-501. doi:10.1038/39057
[7] M. E. Bruce, A. Boyle, S. Cousens, I. McConnell, J. Foster, W. Goldmann and H. Fraser, “Strain Characterization of Natural Sheep Scrapie and Comparison with BSE,” Journal of General Virology, Vol. 83, No. 3, 2002, pp. 695-704.
[8] A. G. Biacabe, J. L. Laplanche, S. Ryder and T. Baron, “Distinct Molecular Phenotypes in Bovine Prion Diseases,” EMBO Reports, Vol. 5, No. 1, 2004, pp. 110-115. doi:10.1038/sj.embor.7400054
[9] A. Buschmann, A. Gretzschel, A. G. Biacabe, K. Schiebel, C. Corona, C. Hoffmann, M. Eiden, T. Baron, C. Casalone and M. H. Groschup, “Atypical BSE in Germany— Proof of Transmissibility and Biochemical Characterization,” Veterinary Microbiology, Vol. 117, No. 2-4, 2006, pp. 103-116. doi:10.1016/j.vetmic.2006.06.016
[10] C. Casalone, G. Zanusso, P. Acutis, S. Ferrari, L. Capucci, F. Tagliavini, S. Monaco and M. Caramelli, “Identification of a Second Bovine Amyloidotic Spongiform Encephalopathy: Molecular Similarities with Sporadic Creutzfeldt-Jakob Disease,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, No. 9, 2004, pp. 3065-3070. doi:10.1073/pnas.0305777101
[11] K. Hagiwara, Y. Yamakawa, Y. Sato, Y. Nakamura, M. Tobiume, M. Shinagawa and T. Sata, “Accumulation of Mono-Glycosylated Form-Rich, Plaque-Forming PrP(Sc) in the Second Atypical Bovine Spongiform Encephalopathy Case in Japan,” Japanese Journal of Infectious Diseases, Vol. 60, No. 5, 2007, pp. 305-308.
[12] J. G. Jacobs, J. P. Langeveld, A. G. Biacabe, P. L. Acutis, M. P. Polak, D. Gavier-Widen, A. Buschmann, M. Caramelli, C. Casalone, M. Mazza, M. Groschup, J. H. Erkens, A. Davidse, F. G. van Zijderveld and T. Baron, “Molecular Discrimination of Atypical Bovine Spongiform Encephalopathy Strains from a Geographical Region Spanning a Wide Area in Europe,” Journal of Clinical Microbiology, Vol. 45, No. 6, 2007, pp. 1821-1829. doi:10.1128/JCM.00160-07
[13] M. P. Polak, J. F. Zmudzinski, J. G. Jacobs and J. P. Langeveld, “Atypical Status of Bovine Spongiform Encephalopathy in Poland: A Molecular Typing Study,” Archives of Virology, Vol. 153, No. 1, 2008, pp. 69-79. doi:10.1007/s00705-007-1062-6
[14] J. A. Richt, R. A. Kunkle, D. Alt, E. M. Nicholson, A. N. Hamir, S. Czub, J. Kluge, A. J. Davis and S. M. Hall, “Identification and Characterization of Two Bovine Spongiform Encephalopathy Cases Diagnosed in the United States,” Journal of Veterinary Diagnostic Investigation, Vol. 19, No. 2, 2007, pp. 142-154. doi:10.1177/104063870701900202
[15] L. A. Terry, R. Jenkins, L. Thorne, S. J. Everest, M. J. Chaplin, L. A. Davis and M. J. Stack, “First Case of H- Type Bovine Spongiform Encephalopathy Identified in Great Britain,” Veterinary Record, Vol. 160, No. 25, 2007, pp. 873-874. doi:10.1136/vr.160.25.873
[16] Y. Yamakawa, K. Hagiwara, K. Nohtomi, Y. Nakamura, M. Nishijima, Y. Higuchi, Y. Sato and T. Sata, “Atypical Proteinase K-Resistant Prion Protein (PrPres) Observed in an Apparently Healthy 23-Month-Old Holstein Steer,” Japanese Journal of Infectious Diseases, Vol. 56, No. 5-6, 2003, pp. 221-222.
[17] D. Gavier-Widen, M. Noremark, J. P. Langeveld, M. Stack, A. G. Biacabe, J. Vulin, M. Chaplin, J. A. Richt, J. Jacobs, C. Acin, E. Monleon, L. Renstrom, B. Klingeborn and T. G. Baron, “Bovine Spongiform Encephalopathy in Sweden: An H-Type Variant,” Journal of Veterinary Diagnostic Investigation, Vol. 20, No. 1, 2008, pp. 2-10. doi:10.1177/104063870802000102
[18] S. Fukuda, Y. Iwamaru, M. Imamura, K. Masujin, Y. Shimizu, Y. Matsuura, Y. Shu, M. Kurachi, K. Kasai, Y. Murayama, S. Onoe, K. Hagiwara, T. Sata, S. Mohri, T. Yokoyama and H. Okada, “Intraspecies Transmission of L-Type-Like Bovine Spongiform Encephalopathy Detected in Japan,” Microbiology and Immunology, Vol. 53, No. 12, 2009, pp. 704-707. doi:10.1111/j.1348-0421.2009.00169.x
[19] Y. Iwamaru, M. Imamura, Y. Matsuura, K. Masujin, Y. Shimizu, Y. Shu, M. Kurachi, K. Kasai, Y. Murayama, S. Fukuda, S. Onoe, K. Hagiwara, Y. Yamakawa, T. Sata, S. Mohri, H. Okada and T. Yokoyama, “Accumulation of L- Type Bovine Prions in Peripheral Nerve Tissues,” Emerging Infectious Diseases, Vol. 16, No. 7, 2010, pp. 1151- 1154. doi:10.3201/eid1607.091882
[20] K. Masujin, Y. Shu, Y. Yamakawa, K. Hagiwara, T. Sata, Y. Matsuura, Y. Iwamaru, M. Imamura, H. Okada, S. Mohri and T. Yokoyama, “Biological and Biochemical Characterization of L-Type-Like Bovine Spongiform Encephalopathy (BSE) Detected in Japanese Black Beef Cattle,” Prion, Vol. 2, No. 3, 2008, pp. 123-128. doi:10.4161/pri.2.3.7437
[21] K. Masujin, R. Miwa, H. Okada, S. Mohri and T. Yokoyama, “Comparative Analysis of Japanese and Foreign L- Type BSE Prions,” Prion, Vol. 6, No. 1, 2012, pp. 89-93. doi:10.4161/pri.6.1.18429
[22] A. Buschmann and M. H. Groschup, “Highly Bovine Spongiform Encephalopathy-Sensitive Transgenic Mice Confirm the Essential Restriction of Infectivity to the Nervous System in Clinically Diseased Cattle,” Journal of Infectious Diseases, Vol. 192, No. 5, 2005, pp. 934- 942. doi:10.1086/431602
[23] V. Béingue, A. Bencsik, A. Le Dur, F. Reine, T. L. Lai, N. Chenais, G. Tilly, A. G. Biacabe, T. Baron, J. L. Vilotte and H. Laude, “Isolation from Cattle of a Prion Strain Distinct from That Causing Bovine Spongiform Encephalopathy,” PLoS Pathogens, Vol. 2, No. 10, 2006, p. e112. doi:10.1371/journal.ppat.0020112
[24] R. Capobianco, C. Casalone, S. Suardi, M. Mangieri, C. Miccolo, L. Limido, M. Catania, G. Rossi, G. Di Fede, G. Giaccone, M. G. Bruzzone, L. Minati, C. Corona, P. Acutis, D. Gelmetti, G. Lombardi, M. H. Groschup, A. Buschmann, G. Zanusso, S. Monaco, M. Caramelli and F. Tagliavini, “Conversion of the BASE Prion Strain into the BSE Strain: The Origin of BSE?” PLoS Pathogens, Vol. 3, No. 3, 2007, p. e31. doi:10.1371/journal.ppat.0030031
[25] T. Kitamoto, S. Mohri, J. W. Ironside, I. Miyoshi, T. Tanaka, N. Kitamoto, S. Itohara, N. Kasai, M. Katsuki, J. Higuchi, T. Muramoto and R. W. Shin, “Follicular Dendritic Cell of the Knock-In Mouse Provides a New Bioassay for Human Prions,” Biochemical and Biophysical Research Communications, Vol. 294, No. 2, 2002, pp. 280- 286. doi:10.1016/S0006-291X(02)00476-X
[26] M. Asano, S. Mohri, J. W. Ironside, M. Ito, N. Tamaoki and T. Kitamoto, “vCJD Prion Acquires Altered Virulence through Trans-Species Infection,” Biochemical and Biophysical Research Communications, Vol. 342, No. 1, 2006, pp. 293-299. doi:10.1016/j.bbrc.2006.01.149
[27] T. Kitamoto, K. Doh-ura, T. Muramoto, M. Miyazono and J. Tateishi, “The Primary Structure of the Prion Protein Influences the Distribution of Abnormal Prion Protein in the Central Nervous System,” American Journal of Pathology, Vol. 141, No. 2, 1992, pp. 271-277.
[28] J. D. Wadsworth, S. Joiner, A. F. Hill, T. A. Campbell, M. Desbruslais, P. J. Luthert and J. Collinge, “Tissue Distribution of Protease Resistant Prion Protein in Variant Creutzfeldt-Jakob Disease Using a Highly Sensitive Immunoblotting Assay,” Lancet, Vol. 358, No. 9277, 2001, pp. 171-180. doi:10.1016/S0140-6736(01)05403-4
[29] T. Muramoto, T. Kitamoto, J. Tateishi and I. Goto, “The Sequential Development of abnormal Prion Protein Accumulation in Mice with Creutzfeldt-Jakob Disease,” American Journal of Pathology, Vol. 140, No. 6, 1992, pp. 1411-1420.
[30] L. McCulloch, K. L. Brown, B. M. Bradford, J. Hopkins, M. Bailey, K. Rajewsky, J. C. Manson and N. A. Mabbott, “Follicular Dendritic Cell-Specific Prion Protein (PrP) Expression Alone Is Sufficient to Sustain Prion Infection in the Spleen,” PLoS Pathogens, Vol. 7, No. 12, 2011, Article ID: e1002402. doi:10.1371/journal.ppat.1002402
[31] B. E. Schreuder and R. A. Somerville, “Bovine Spongiform Encephalopathy in Sheep?” Revue Scientifique et Technique, Vol. 22, No. 1, 2003, pp. 103-120.

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.