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TSE strain differentiation in mice by immunohistochemical PrP Sc profiles and triplex W estern blot
Author(s) -
Keulen Lucien J. M.,
Langeveld Jan P. M.,
Dolstra Corry H.,
Jacobs Jorg,
Bossers Alex,
Zijderveld Fred G.
Publication year - 2015
Publication title -
neuropathology and applied neurobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.538
H-Index - 95
eISSN - 1365-2990
pISSN - 0305-1846
DOI - 10.1111/nan.12181
Subject(s) - western blot , microbiology and biotechnology , biology , blot , immunohistochemistry , strain (injury) , epitope , inbred strain , antibody , genetics , anatomy , immunology , gene
TSE strains are routinely identified by their incubation period and vacuolation profile in the brain after intracerebral inoculation and serial passaging in inbred mouse lines. There are some major drawbacks to this method that are related to the variation in vacuolation that exists in the brains of mice infected with the same TSE strain and to variation between observers and laboratories in scoring vacuolation and determining the final incubation period. Aim We investigated the potential of PrP Sc immunohistochemistry and triplex Western blotting as possible alternative methods to differentiate between TSE strains. Methods TSE reference strains ME 7, 87 A /87 V , 22 A /22 C , 79 A /79 V and 301 C /301 V were intracerebrally inoculated in RIII or VM inbred mice that differ in their PrP genotype. Immunohistochemical PrP Sc profiles were drawn up by scanning light microscopy both on coronal and sagittal sections. Results On the basis of the localization of PrP Sc in the cerebral cortex, hippocampus, and cerebellar cortex and the overall type of PrP Sc staining, all TSE strains could be well differentiated from each other through their typical strain dependent characteristics. In addition, Western blot showed that the combination of glycosylation profile and 12 B 2 epitope content of PrP Sc allowed to distinguish between all reference strains except for ME 7 and 22 A in VM mice. Conclusion TSE strains in mice can be identified on the basis of their PrP Sc profile alone. The potential to identify TSE strains in ruminants with these PrP Sc profiles after a single primary passage in mice will be the topic of future studies.