Physical, chemical and kinetic factors affecting prion infectivity.
The mouse-adapted scrapie prion strain RML is one of the most widely used in prion research. The introduction of a cell culture-based assay of RML prions, the scrapie cell assay (SCA) allows more rapid and precise prion titration. A semi-automated version of this assay (ASCA) was applied to explore a range of conditions that might influence the infectivity and properties of RML prions. These include resistance to freeze-thaw procedures; stability to endogenous proteases in brain homogenate despite prolonged exposure to varying temperatures; distribution of infective material between pellet and supernatant after centrifugation, the effect of reducing agents and the influence of detergent additives on the efficiency of infection. Apparent infectivity is increased significantly by interaction with cationic detergents. Importantly, we have also elucidated the relationship between the duration of exposure of cells to RML prions and the transmission of infection. We established that the infection process following contact of cells with RML prions is rapid and followed an exponential time course, implying a single rate-limiting process.
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Francesca
F
Properzi
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Anjna
A
Badhan
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Steffi
S
Klier
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Christian
C
Schmidt
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Peter C
PC
Klöhn
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Jonathan D F
JD
Wadsworth
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Anthony R
AR
Clarke
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
Graham S
GS
Jackson
a MRC Prion Unit, Department of Neurodegenerative Disease , UCL Institute of Neurology , London , UK.
John
J
Collinge
D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology; D018929:Cell Culture Techniques; D002460:Cell Line; D003902:Detergents / Q000378:metabolism; D005615:Freezing; D007700:Kinetics; D051379:Mice; D011328:Prions / Q000032:analysis / Q000378:metabolism* / Q000472:pathogenicity*; D019163:Reducing Agents / Q000378:metabolism; D012608:Scrapie / Q000378:metabolism* / Q000473:pathology / Q000635:transmission*; D013696:Temperature
D003902:Detergents; D011328:Prions; D019163:Reducing Agents
BSE; CJD; PrP; RML; cell culture; prion; scrapie