Limits...
Intraperitoneal Infection of Wild-Type Mice with Synthetically Generated Mammalian Prion.

Wang X, McGovern G, Zhang Y, Wang F, Zha L, Jeffrey M, Ma J - PLoS Pathog. (2015)

Bottom Line: However, whether the pathogenic properties of synthetically generated prion (rec-Prion) recapitulate those of naturally occurring prions remains unresolved.Detailed pathological analyses revealed that the nature of rec-Prion induced lesions, including spongiform change, disease specific prion protein accumulation (PrP-d) and the PrP-d dissemination amongst lymphoid and peripheral nervous system tissues, the route and mechanisms of neuroinvasion were all typical of classical rodent prions.Our results revealed that, similar to naturally occurring prions, the rec-Prion has a titratable infectivity and is capable of causing prion disease via routes other than direct intra-cerebral challenge.

View Article: PubMed Central - PubMed

Affiliation: Center for Neurodegenerative Science, Van Andel Research Institute, Grand Rapids, Michigan, United States of America; Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, Ohio, United States of America.

ABSTRACT
The prion hypothesis postulates that the infectious agent in transmissible spongiform encephalopathies (TSEs) is an unorthodox protein conformation based agent. Recent successes in generating mammalian prions in vitro with bacterially expressed recombinant prion protein provide strong support for the hypothesis. However, whether the pathogenic properties of synthetically generated prion (rec-Prion) recapitulate those of naturally occurring prions remains unresolved. Using end-point titration assay, we showed that the in vitro prepared rec-Prions have infectious titers of around 104 LD50/μg. In addition, intraperitoneal (i.p.) inoculation of wild-type mice with rec-Prion caused prion disease with an average survival time of 210-220 days post inoculation. Detailed pathological analyses revealed that the nature of rec-Prion induced lesions, including spongiform change, disease specific prion protein accumulation (PrP-d) and the PrP-d dissemination amongst lymphoid and peripheral nervous system tissues, the route and mechanisms of neuroinvasion were all typical of classical rodent prions. Our results revealed that, similar to naturally occurring prions, the rec-Prion has a titratable infectivity and is capable of causing prion disease via routes other than direct intra-cerebral challenge. More importantly, our results established that the rec-Prion caused disease is pathogenically and pathologically identical to naturally occurring contagious TSEs, supporting the concept that a conformationally altered protein agent is responsible for the infectivity in TSEs.

No MeSH data available.


Related in: MedlinePlus

Vacuolar lesion profile of CD-1 mice challenged by i.p. inoculation of rec-Prions.Spongiosis in the following brain regions were scored. Grey matter regions: G1, dorsal medulla; G2, cerebellar cortex; G3, superior colliculus; G4, hypothalamus; G5, medial thalamus; G6, hippocampus; G7, septum; G8, parietal cerebral cortex; G9, frontal cerebral cortex. White matter regions: W1, cerebellar white matter; W2, mesencephalic tegmentum; W3, pyramidal tract. Error bars represent SEM (standard error of the mean).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4489884&req=5

ppat.1004958.g003: Vacuolar lesion profile of CD-1 mice challenged by i.p. inoculation of rec-Prions.Spongiosis in the following brain regions were scored. Grey matter regions: G1, dorsal medulla; G2, cerebellar cortex; G3, superior colliculus; G4, hypothalamus; G5, medial thalamus; G6, hippocampus; G7, septum; G8, parietal cerebral cortex; G9, frontal cerebral cortex. White matter regions: W1, cerebellar white matter; W2, mesencephalic tegmentum; W3, pyramidal tract. Error bars represent SEM (standard error of the mean).

Mentions: The ability to infect mice via the i.p route allows us to determine the pathogenic changes in rec-Prion caused disease, including disease pathology in central nervous system (CNS) and lymphoreticular system (LRS), and the route and mechanism of neuroinvasion. Detailed pathological analyses and a time course study were performed (Table 2 experiment 5), which allows us to compare the i.p. rec-Prion challenge induced disease to that of classical rodent scrapie. Figs 3 and 4 shows the patterns of vacuolation and immunohistochemically detected aberrant PrP deposition (PrP-d, the disease specific PrP form detected by histological methods) in terminally diseased mice. PrP-d in CNS and spinal cord was mainly found as diffuse punctuate forms with frequent granular intra-cellular accumulations (Fig 4) and occasional small plaque-like deposits. Thus, both the nature and patterns of vacuolation and PrP-d accumulation are consistent with those found in different strains and isolates of laboratory murine scrapie and supports the contention that rec-Prions causes bona fide prion disease in rodents [31].


Intraperitoneal Infection of Wild-Type Mice with Synthetically Generated Mammalian Prion.

Wang X, McGovern G, Zhang Y, Wang F, Zha L, Jeffrey M, Ma J - PLoS Pathog. (2015)

Vacuolar lesion profile of CD-1 mice challenged by i.p. inoculation of rec-Prions.Spongiosis in the following brain regions were scored. Grey matter regions: G1, dorsal medulla; G2, cerebellar cortex; G3, superior colliculus; G4, hypothalamus; G5, medial thalamus; G6, hippocampus; G7, septum; G8, parietal cerebral cortex; G9, frontal cerebral cortex. White matter regions: W1, cerebellar white matter; W2, mesencephalic tegmentum; W3, pyramidal tract. Error bars represent SEM (standard error of the mean).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4489884&req=5

ppat.1004958.g003: Vacuolar lesion profile of CD-1 mice challenged by i.p. inoculation of rec-Prions.Spongiosis in the following brain regions were scored. Grey matter regions: G1, dorsal medulla; G2, cerebellar cortex; G3, superior colliculus; G4, hypothalamus; G5, medial thalamus; G6, hippocampus; G7, septum; G8, parietal cerebral cortex; G9, frontal cerebral cortex. White matter regions: W1, cerebellar white matter; W2, mesencephalic tegmentum; W3, pyramidal tract. Error bars represent SEM (standard error of the mean).
Mentions: The ability to infect mice via the i.p route allows us to determine the pathogenic changes in rec-Prion caused disease, including disease pathology in central nervous system (CNS) and lymphoreticular system (LRS), and the route and mechanism of neuroinvasion. Detailed pathological analyses and a time course study were performed (Table 2 experiment 5), which allows us to compare the i.p. rec-Prion challenge induced disease to that of classical rodent scrapie. Figs 3 and 4 shows the patterns of vacuolation and immunohistochemically detected aberrant PrP deposition (PrP-d, the disease specific PrP form detected by histological methods) in terminally diseased mice. PrP-d in CNS and spinal cord was mainly found as diffuse punctuate forms with frequent granular intra-cellular accumulations (Fig 4) and occasional small plaque-like deposits. Thus, both the nature and patterns of vacuolation and PrP-d accumulation are consistent with those found in different strains and isolates of laboratory murine scrapie and supports the contention that rec-Prions causes bona fide prion disease in rodents [31].

Bottom Line: However, whether the pathogenic properties of synthetically generated prion (rec-Prion) recapitulate those of naturally occurring prions remains unresolved.Detailed pathological analyses revealed that the nature of rec-Prion induced lesions, including spongiform change, disease specific prion protein accumulation (PrP-d) and the PrP-d dissemination amongst lymphoid and peripheral nervous system tissues, the route and mechanisms of neuroinvasion were all typical of classical rodent prions.Our results revealed that, similar to naturally occurring prions, the rec-Prion has a titratable infectivity and is capable of causing prion disease via routes other than direct intra-cerebral challenge.

View Article: PubMed Central - PubMed

Affiliation: Center for Neurodegenerative Science, Van Andel Research Institute, Grand Rapids, Michigan, United States of America; Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, Ohio, United States of America.

ABSTRACT
The prion hypothesis postulates that the infectious agent in transmissible spongiform encephalopathies (TSEs) is an unorthodox protein conformation based agent. Recent successes in generating mammalian prions in vitro with bacterially expressed recombinant prion protein provide strong support for the hypothesis. However, whether the pathogenic properties of synthetically generated prion (rec-Prion) recapitulate those of naturally occurring prions remains unresolved. Using end-point titration assay, we showed that the in vitro prepared rec-Prions have infectious titers of around 104 LD50/μg. In addition, intraperitoneal (i.p.) inoculation of wild-type mice with rec-Prion caused prion disease with an average survival time of 210-220 days post inoculation. Detailed pathological analyses revealed that the nature of rec-Prion induced lesions, including spongiform change, disease specific prion protein accumulation (PrP-d) and the PrP-d dissemination amongst lymphoid and peripheral nervous system tissues, the route and mechanisms of neuroinvasion were all typical of classical rodent prions. Our results revealed that, similar to naturally occurring prions, the rec-Prion has a titratable infectivity and is capable of causing prion disease via routes other than direct intra-cerebral challenge. More importantly, our results established that the rec-Prion caused disease is pathogenically and pathologically identical to naturally occurring contagious TSEs, supporting the concept that a conformationally altered protein agent is responsible for the infectivity in TSEs.

No MeSH data available.


Related in: MedlinePlus