NR AXXY
AU Tsukui,K.; Takata,M.; Tadokoro,K.; Onodera,T.
TI A Potential Blood Test for TSE by Detecting Carbohydrate-dependent Aggregates of PrPres-like Proteins in Scrapie-infected Hamster Plasma
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Epidemiology, Risk Assessment and Transmission P04.57
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: PrPres has rarely been detected in blood, except in leukocytes, even in diseased animal models which show a large amount of PrPres in affected organs. It seems likely that PrPres detection in blood is difficult because of the low titer of infectivity of the material in blood.
Aims/Objectives: To demonstrate the detection of proteinase K-resistant 3F4-reactive protein in the plasma of scrapie-infected hamsters but not in the plasma of mock infected hamsters.
Methods: Partial purification using a novel method termed acidic SDS precipitation and a highly sensitive chemiluminescence detection system after conventional western blotting was used.
Results: The chemiluminescence method could show the presence of PrPc at a concentration equivalent to 1.4x10-9g of brain homogenate or 1.5x10-12 g (6.5x10-17 mole) of rHaPrP. Using the above method, the 3F4-reactive proteins in scrapie-infected hamster plasma were often indicated as multiple Mw protein bands between higher Mw positions than position of di-glycosyl PrP molecule. Mixing of scrapie-infected hamster brain homogenate with mock infected hamster plasma resulted in the formation of similar Mw of multiple 3F4-reactive proteins. Predigestion of carbohydrate chains from the proteins in plasma or brain homogenate before mixing resulted in the failure to obtain these multiple 3F4-reactive proteins.
Discussion: These observations sugested that PrPres was aggregated with self or other protein molecules in plasma through carbohydrate side chains. This type of PrPres was successfully detected in the plasma of scrapie-infected hamster. Biological properties of these aggregates with PrPres-like protein in scHaPl are not known.
Acknowledgement: We gratefully acknowledge Dr. Takashi Yokoyama, Research Institute for Prion Diseases, National Institute of animal Health of Japan, for his greatest support to use scrapie-infected hamster materials.
AD K. Tsukui, K. Tadokoro, Central Blood Institute of the Japanese Red Cross Society, Japan; M. Takata, National Institute of Animal Health Japan, Research Center for Prion Diseases, Japan; T. Onodera, Agriculture and Life Sciences, Tokyo University, Department of Molecular Immunology, Japan
SP englisch
PO Schottland