NR AXSZ
AU Morales,R.; Buytaert-Hoefen,K.; Hansen,E.; Hlavinka,D.; Goodrich,R.; Soto,C.
TI Reduction of Prion Infectivity in Blood by Navigant Prototype Separation and Cell Washing Device
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Epidemiology, Risk Assessment and Transmission P04.53
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB Although prion diseases are rare in humans, the established link between a new variant form of CJD (vCJD) and the consumption of cattle meat contaminated by BSE have raised concern about a possible outbreak of a large epidemic in the human population. Over the past few years, BSE has become a significant health problem affecting many countries, and it seems now apparent that vCJD can be iatrogenically transmitted from human to human by blood transfusion. Exacerbating this state of affairs is the lack of a reliable test to identify individuals incubating the disease during the long and silent period from the onset of infection to the appearance of clinical symptoms. The purpose of this research study was to evaluate the effectiveness of the Navigant Prototype separation and cell washing device to remove infectious scrapie prion protein (PrPsc) from red blood cell (RBC) suspensions in prion spiked samples. Analysis of the treated sample supernatants by Western blot revealed that approximately 88% of the PrPsc was removed with the initial plasma expression and the equivalent to 6% was detected in a saline wash. The final sample of RBCs revealed no detectable levels of PrPsc by western blots, but by extrapolation of the removal rate, we calculate that the Navigant blood separation and cell washing device removed 98.5% PrPsc. Further analysis of the treated RBCs using the PMCA assay indicated detectable amounts of PrPsc only after 2 consecutive amplification rounds. Semi-quantitative analysis of PMCA amplification enabled us to estimate that the treated RBCs contained less than 1 x 104 LD50 PrPsc. These in vitro estimations were confirmed by in vivo infectivity studies. Preliminary in vivo data displayed significant differences in the incubation periods of the spiked blood inoculated hamsters (100.1 ± 1.7) versus washed RBCs (135.8 ± 6.7 with an incomplete attack rate). Moreover, a substantial difference in the attack rate (40% in RBC, versus 100% in spiked blood) further indicates a substantial removal of infectious prions. Comparison of this data with results of animals inoculated with different dilutions of infectious material, indicate a >99.0% reduction of infectivity. This data suggest that the Navigant separation and cell washing device represents an efficient method to remove prions from blood fractions and its application may lead to increased safety of blood products prepared in this way.
AD R. Morales, C. Soto, University of Texas Medical Branch, Neurology, USA; K. Buytaert-Hoefen, E. Hansen, D. Hlavinka, R. Goodrich, Navigant Biotechnologies, USA
SP englisch
PO Schottland