Preparation of soluble infectious samples from scrapie-infected brain: a new tool to study the clearance of transmissible spongiform encephalopathy agents during plasma fractionation

BACKGROUND:  Concern about the safety of blood, blood components, and plasma‐derived products with respect to prions has increased since the report of two blood‐related infections of variant Creutzfeldt‐Jakob disease in the United Kingdom. Efforts were directed toward the development of procedures a...

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Bibliographic Details
Published in:Transfusion (Philadelphia, Pa.) Pa.), 2006-04, Vol.46 (4), p.652-658
Main Authors: Berardi, Vito Angelo, Cardone, Franco, Valanzano, Angelina, Lu, Mei, Pocchiari, Maurizio
Format: Article
Language:English
Subjects:
Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Animals
Biological and medical sciences
Blood Component Transfusion - adverse effects
Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis
Brain - pathology
Cricetinae
Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases
Injuries of the nervous system and the skull. Diseases due to physical agents
Medical sciences
Mesocricetus
Models, Animal
Neurology
Prion Diseases - blood
Prion Diseases - prevention & control
Prion Diseases - transmission
Prions - isolation & purification
PrPSc Proteins - isolation & purification
Transfusions. Complications. Transfusion reactions. Cell and gene therapy
Traumas. Diseases due to physical agents
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Summary:BACKGROUND:  Concern about the safety of blood, blood components, and plasma‐derived products with respect to prions has increased since the report of two blood‐related infections of variant Creutzfeldt‐Jakob disease in the United Kingdom. Efforts were directed toward the development of procedures able to remove or inactivate prions from blood components or plasma‐derived products with brain fractions of transmissible spongiform encephalopathy (TSE)‐infected rodents as spiking materials. These spiking materials, however, are loaded with pathological prion protein (PrPTSE) aggregates that are likely not associated to blood infectivity. The presence of these aggregates may invalidate these studies. STUDY DESIGN AND METHODS:  Brains from 263K scrapie–infected hamsters were suspended in 10 percent phosphate‐buffered saline. After low‐speed centrifugation, the supernatant was collected and ultracentrifuged at 220,000 × g at 25°C for 30 minutes. The high‐speed supernatants (SHS) and pellets were collected; the proteinase‐resistant PrPTSE was measured by Western blot and infectivity by intracerebral inoculation into weanling hamsters. RESULTS:  A substantial amount of prion infectivity (more than 105 LD50 per mL of a 10% suspension of brain tissues) is present in the SHS fraction of 263K scrapie–infected hamster brains. Concomitantly, this fraction contains none or only traces of PrPTSE in its aggregate form. CONCLUSION:  This study describes a simple and fast protocol to prepare infectious material from 263K scrapie–infected brains that is not contaminated with PrPTSE aggregates. This SHS fraction is likely to be the most relevant material for endogenous spiking of human blood in validation experiments aimed at demonstrating procedures to remove or inactivate TSE infectious agents.
ISSN:0041-1132
1537-2995
DOI:10.1111/j.1537-2995.2006.00763.x