NR AWKP
AU Morales,R.; Castilla,J.; Saa,P.; Soto,C.
TI In vitro propagation of prion strains
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PR-24
PT Konferenz-Poster
AB Prions are unconventional infectious agents responsible for transmissible spongiform encephalopathies. Compelling evidence support the hypothesis that prions are composed exclusively by a misfolded form of the prion protein (PrPsc) that replicates in the absence of nucleic acids. It is well established that the prion infectious agent, like conventional micro-organisms, exhibit strain variation. Prion strains can be differentiated by their particular in vivo and in vitro characteristics, including differences in incubation period, lesion pattern of infected brains and biochemical properties of PrPsc, among others. Understanding how a single protein can provide the diversity to sustain the strain phenomenon has been a challenge for the prion hypothesis. We have recently described that PrPsc can be propagated indefinitely in vitro to generate infectious material using the protein misfolding cyclic amplification (PMCA) technology. Here we demonstrate that prion strains characteristics can be replicated in vitro by PMCA. Our results show in vitro amplification of various mouse strains including 301C, RML, ME7, 139A, and 79A. Moreover, wild type mice inoculated intracerebrally with in vitro generated PrPsc from 301C, RML and 79A strains show clinical signs, brain lesion pattern and biochemical characteristics identical to the animals inoculated with brain infectious material. Interestingly, the specific infectivity (infectivity per unit of PrPsc) was the same in both preparations. These results suggest that the strain characteristics of prions can be maintained by in vitro replication of PrPsc.
AD R. Morales: Department of Neurology, University of Texas Medical Branch, Galveston TX 77550 and Facultad de Ciencias, Universidad de Chile, Santiago, Chile; J. Castilla, C. Soto: Department of Neurology, University of Texas Medical Branch, Galveston TX 77550; P. Saá: Department of Neurology, University of Texas Medical Branch, Galveston TX 77550 and Centro de Biología Molecular, Universidad Autónoma de Madrid, Madrid, Spain
SP englisch
PO Italien