NR AMAS
AU Vilette,D.; Andreoletti,O.; Archer,F.; Madelaine,M.F.; Vilotte,J.L.; Lehmann,S.; Laude,H.
TI Ex vivo propagation of infectious sheep scrapie agent in heterologous epithelial cells expressing ovine prion protein
QU Proceedings of the National Academy of Sciences of the United States of America 2001 Mar 27; 98(7): 4055-9
IA http://www.pnas.org/cgi/content/full/98/7/4055
PT journal article
AB Transmissible spongiform encephalopathies, or prion diseases, are fatal degenerative disorders of the central nervous system that affect humans and animals. Prions are nonconventional infectious agents whose replication depends on the host prion protein (PrP). Transmission of prions to cultured cells has proved to be a particularly difficult task, and with a few exceptions, their experimental propagation relies on inoculation to laboratory animals. Here, we report on the development of a permanent cell line supporting propagation of natural sheep scrapie. This model was obtained by stable expression of a tetracycline-regulatable ovine PrP gene in a rabbit epithelial cell line. After exposure to scrapie agent, cultures were repeatedly found to accumulate high levels of abnormal PrP (PrPres). Cell extracts induced a scrapie-like disease in transgenic mice overexpressing ovine PrP. These cultures remained healthy and stably infected upon subpassaging. Such data show that (i) cultivated cells from a nonneuronal origin can efficiently replicate prions; and (ii) species barrier can be crossed ex vivo through the expression of a relevant PrP gene. This approach led to the ex vivo propagation of a natural transmissible spongiform encephalopathy agent (i.e., without previous experimental adaptation to rodents) and might be applied to human or bovine prions.
MH Animal; Cells, Cultured; Epithelial Cells/*metabolism; Mice; PrPsc Proteins/*biosynthesis; Prion Diseases/metabolism/*veterinary; Prions/*biosynthesis/genetics; Rabbits; Sheep; Sheep Diseases/*metabolism; Support, Non-U.S. Gov't; Variation (Genetics)
AD vilette@biotec.jouy.inra.fr ; Unite de Virologie Immunologie Moleculaires and Laboratoire de Genetique Biochimique et Cytogenetique, Institut National de la Recherche Agronomique, 78350 Jouy-en-Josas, France; UMR Institut National de la Recherche Agronomique-Ecole Nationale Veterinaire de Toulouse 959 de Physiopathologie Infectieuse et Parasitaire des Ruminants, Ecole Nationale Veterinaire de Toulouse, 31076 Toulouse, France; and Institut de Genetique Humaine, Centre National de la Recherche Scientifique, 34396 Montpellier, France.
SP englisch
PO USA