In vitro Interaction of Scrapie Agent and Mouse Peritoneal Macrophages

Carp, Richard I.; Callahan, Sharon M.

doi:10.1159/000149241

Cited by 58 publications

(40 citation statements)

References 9 publications

(10 reference statements)

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“…2). This is in agreement with previous studies by Carp & Callahan (1981, 1982 who showed that scrapie agent was associated with macrophages following incubation in vitro for 2 h at 37 °C. They then showed that with longer incubation in vitro, scrapie infectivity decreased as measured by an increase in incubation period (Carp & Callahan, 1982).…”

Section: Discussionsupporting

confidence: 94%

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In vitro Replication of Scrapie Agent in a Neuronal Model: Infection of PC12 Cells

Rubenstein

Carp

Callahan

1984

Journal of General Virology

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SUMMARYA rat phaeochromocytoma cell line, termed PC12, was used to study scrapie replication. These cells, in response to the addition of nerve growth factor (NGF), exhibit a number of neuronal properties including morphological differentiation, electrophysiological responsiveness, and neurotransmitter synthesis. Cultures were exposed to scrapie brain homogenate (strain 139A), harvested every week for up to 6 weeks, and assayed for scrapie infectivity. Scrapie replication in vitro was monitored by injecting scrapie agent-exposed NGF-treated PC 12 cells into mice and measuring time intervals from injection to onset of clinical symptoms. Mouse incubation periods vary inversely with the amount of scrapie infectivity present. Cells harvested at 7 and 14 days after exposure to scrapie agent showed a decrease in the level of infectivity followed by an increase at subsequent time points. The increase in scrapie infectivity from early to late time intervals after agent exposure clearly indicated replication in vitro. A fusion agent was not necessary to establish infection, and the addition of mouse peritoneal macrophages caused a reduction in the yield of infectivity per culture. Examination of cells by phase-contrast microscopy failed to reveal any cytopathology.

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Section: Discussionsupporting

confidence: 94%

“…injection of 2.0 to 3.0 ml of thioglycolate. Macrophages were harvested as previously described (Carp & Callahan, 1981) and seeded 1 week prior to exposure to agent. Cells (3 × l04 to 5 × 104) were plated on collagen-coated 35 mm Petri dishes in maintenance medium in the presence or absence of NGF-treated PC12 cells.…”

Section: Methodsmentioning

confidence: 99%

In vitro Replication of Scrapie Agent in a Neuronal Model: Infection of PC12 Cells

Rubenstein

Carp

Callahan

1984

Journal of General Virology

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“…The scrapie agent 263K was inoculated into LVG/LAK hamsters. The inoculum preparation, intracerebral injections, scoring and sacrificing of animals were performed as previously described (Carp & Callahan, 1981). Control animals were injected with normal brain homogenate.…”

Section: Methodsmentioning

confidence: 99%

Detection of Scrapie-associated Fibril (SAF) Proteins Using Anti-SAF Antibody in Non-purified Tissue Preparations

Rubenstein

Kascsak

Merz

et al. 1986

Journal of General Virology

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SUMMARY Antisera raised to scrapie-associated fibril (SAF) proteins were used to detect scrapie-specific polypeptides in three different non-purified brain preparations: a synaptosomal-mitochondrial fraction, 20% brain homogenate and 20% brain homogenate extracted with Sarkosyl. The concentration of SAF proteins in the preparations was greater than the quantity of SAF as detected by negative stain electron microscopy. This suggests that not all of the protein exists in the form of SAF. An immunologically reactive 33K to 35K protein was detected in both normal and scrapie brain preparations. This protein was susceptible to complete proteinase K (PK) digestion in normal brain preparations and it is suggested that scrapie infection is responsible for post-translational modifications which confer PK resistance in scrapie preparations. These modifications may also play a role in the antigenic differences seen in a variety of scrapie agents. SAF-specific proteins were also detected in the spinal cords and spleens from scrapie-affected animals. Detergent extraction of material followed by PK treatment and Western blot analysis is a highly specific and sensitive method for the detection of SAF proteins. This procedure could be applied to human neurological diseases of unknown aetiology.

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“…Scrapie infectivity has been reported to associate with macrophages (Lavelle et al, 1972;Carp & Callahan, 1981;Marsh, 1981) and certainly they are likely candidates for a role in uptake, degradation and processing of the inoculum, and possibly in the early cell-to-cell transmission of infectivity when titres are increasing in lymphoreticular sites such as the spleen. This is particularly relevant because in the crude inocula used the infective units are closely associated with the tissue debris.…”

Section: F Farquhar and A G Dickinsonmentioning

confidence: 99%

Prolongation of Scrapie Incubation Period by an Injection of Dextran Sulphate 500 within the Month Before or After Infection

Farquhar

Dickinson

1986

Journal of General Virology

125

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In vitro Interaction of Scrapie Agent and Mouse Peritoneal Macrophages

Cited by 58 publications

References 9 publications

In vitro Replication of Scrapie Agent in a Neuronal Model: Infection of PC12 Cells

In vitro Replication of Scrapie Agent in a Neuronal Model: Infection of PC12 Cells

Detection of Scrapie-associated Fibril (SAF) Proteins Using Anti-SAF Antibody in Non-purified Tissue Preparations

Prolongation of Scrapie Incubation Period by an Injection of Dextran Sulphate 500 within the Month Before or After Infection

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