Evidence for the Multiplication of Scrapie Agent in Cell Culture

Clarke, Martin; Haig, D.A.

doi:10.1038/225100a0

Cited by 99 publications

(67 citation statements)

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“…Prion replication in neuronal cell lines was first demonstrated in 1970 [36] and since then several studies have established that prion inoculation of certain neuronal cell lines results in propagation of infectivity and accumulation of the pathological isoform of the prion protein, PrP Sc [37,38]. One of the most commonly used cell line for in vitro prion studies is the mouse neuroblastoma cell line N2a which is readily susceptible to mouse adapted sheep scrapie prion strains.…”

Section: Page | 24mentioning

confidence: 99%

Canine MDCK cell lines are refractory to infection with human and mouse prions

Polymenidou

Trusheim

Stallmach

et al. 2008

Vaccine

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Section: Page | 24mentioning

confidence: 99%

Canine MDCK cell lines are refractory to infection with human and mouse prions

Polymenidou

Trusheim

Stallmach

et al. 2008

Vaccine

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“…PC12 rat pheochromocytoma cells can be differentiated into neuron-like cells which can then be infected with mouse prions [118]. SMB cells were originally isolated from Chandler strain-infected mouse brain culture [31]. However, Chandler-infected SMB cells can be cured with pentosan sulphate and infected de novo with other mouse-adapted prion strains [13,65].…”

Section: Cell Models Permissive To Experimental Rodent-adapted Scrapmentioning

confidence: 99%

“…The first successful attempt to establish scrapie-infected cultures was obtained as early as 1970 when low levels of prion multiplication was reported in a mouse brain-derived cell line called SMB [31]. Since then, and despite different repeated attempts to maintain scrapie-infected cultures [114], stable and consistent propagation of prions was achieved in a very limited number of cell cultures.…”

Section: Introductionmentioning

confidence: 99%

Cell models of prion infection

Vilette

2007

Vet. Res.

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-Due to recent renewal of interest and concerns in prion diseases, a number of cell systems permissive to prion multiplication have been generated in the last years. These include established cell lines, neuronal stem cells and primary neuronal cultures. While most of these models are permissive to experimental, mouse-adapted strains of prions, the propagation of natural field isolates from sheep scrapie and chronic wasting disease has been recently achieved. These models have improved our knowledge on the molecular and cellular events controlling the conversion of the PrP C protein into abnormal isoforms and on the cell-to-cell spreading of prions. Infected cultured cells will also facilitate investigations on the molecular basis of strain identity and on the mechanisms that lead to neurodegeneration. The ongoing development of new cell models with improved characteristics will certainly be useful for a number of unanswered critical issues in the prion field.

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“…In the former case, it has been established that immortalized cell groups are generated in prion-infected animals. Thus, primary cultures from prion-infected brains can easily provide cells with continuous passage, i.e., ScHB and SMB produced from prion-infected animals [19]. Several cell lines represent the examples of the latter case.…”

Section: Prion Persistently Infected and Suscepti-ble Cell Linesmentioning

confidence: 99%

Recent Developments in Prion Disease Research: Diagnostic Tools and In Vitro Cell Culture Models

Sakudo

Nakamura

Ikuta

et al. 2007

The Journal of Veterinary Medical Science

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ABSTRACT. After prion infection, an abnormal isoform of prion protein (PrP Sc ) converts the cellular isoform of prion protein (PrP C ) into PrP Sc . PrP C -to-PrP Sc conversion leads to PrP Sc accumulation and PrP C deficiency, contributing etiologically to induction of prion diseases. Presently, most of the diagnostic methods for prion diseases are dependent on PrP Sc detection. Highly sensitive/accurate specific detection of PrP Sc in many different samples is a prerequisite for attempts to develop reliable detection methods. Towards this goal, several methods have recently been developed to facilitate sensitive and precise detection of PrP Sc , namely, protein misfolding cyclic amplification, conformation-dependent immunoassay, dissociation-enhanced lanthanide fluorescent immunoassay, capillary gel electrophoresis, fluorescence correlation spectroscopy, flow microbead immunoassay, etc. Additionally, functionally relevant prion-susceptible cell culture models that recognize the complexity of the mechanisms of prion infection have also been pursued, not only in relation to diagnosis, but also in relation to prion biology. Prion protein (PrP) gene-deficient neuronal cell lines that can clearly elucidate PrP C functions would contribute to understanding of the prion infection mechanism. In this review, we describe the trend in recent development of diagnostic methods and cell culture models for prion diseases and their potential applications in prion biology. Evidence of prion infections has been established by the accumulation of an abnormal isoform of prion protein (PrP Sc ) and/or infectivity after multiple passages [54,85,94,131]. In vitro conversion of the cellular isoform of prion protein (PrP C ) to form PrP Sc -like products has been demonstrated by incubating 35 S-labeled PrP C with PrP Sc . This produced a protease-resistant radioactive product with the mobility of protease-treated authentic PrP Sc [55]. This in vitro conversion confirms the species-[90] and strain-specificities [7] observed in vivo. However, because the yield is less stoichiometric than for PrP Sc [55], it has not been possible to determine whether or not an increase in infectivity occurred [55]. Recent progress in the development of diagnostic methods for PrP Sc detection has dramatically facilitated many approaches in prion biology. The protein misfolding cyclic amplification (PMCA) method deserves special mention [96]. Hitherto, it has not been possible to renature completely denatured prion preparations to an infectious state [79,81]; however, the novel PMCA approach enables in vitro amplification of infectious prions [16].The susceptibility of cell lines, such as N2a, in an in vitro cell culture assay to prion infections has been assessed by exposing the cells to serial dilutions of prion-infected mouse brain homogenates, and the dilution rate yielding negative outcome in a blotting assay for PrP Sc has been determined to be the infectivity level. Prion concentrations equivalent to those determined in an animal bioassay [9] have ...

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Evidence for the Multiplication of Scrapie Agent in Cell Culture

Cited by 99 publications

References 1 publication

Canine MDCK cell lines are refractory to infection with human and mouse prions

Canine MDCK cell lines are refractory to infection with human and mouse prions

Cell models of prion infection

Recent Developments in Prion Disease Research: Diagnostic Tools and In Vitro Cell Culture Models

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