Lower specific infectivity of protease-resistant prion protein generated in cell-free reactions

被引：50

作者：

Klingeborn, Mikael ^{[1
]}

Race, Brent ^{[1
]}

Meade-White, Kimberly D. ^{[1
]}

Chesebro, Bruce ^{[1
]}

机构：

[1] NIAID, Persistent Viral Dis Lab, Rocky Mt Labs, Hamilton, MT 59840 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2011年 / 108卷 / 48期

关键词：

Creutzfeldt-Jakob disease; amyloid; neurodegeneration; protein polymerization; protein aggregation; CHRONIC WASTING DISEASE; IN-VITRO; SPONGIFORM ENCEPHALOPATHIES; SCRAPIE AGENT; CYCLIC AMPLIFICATION; INCUBATION PERIOD; TRANSGENIC MICE; BRAIN; ASSAY; REPLICATION;

D O I：

10.1073/pnas.1111255108

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Prions are unconventional infectious agents that cause transmissible spongiform encephalopathy (TSE) diseases, or prion diseases. The biochemical nature of the prion infectious agent remains unclear. Previously, using a protein misfolding cyclic amplification (PMCA) reaction, infectivity and disease-associated protease-resistant prion protein (PrPres) were both generated under cell-free conditions, which supported a nonviral hypothesis for the agent. However, these studies lacked comparative quantitation of both infectivity titers and PrPres, which is important both for biological comparison with in vivo-derived infectivity and for excluding contamination to explain the results. Here during four to eight rounds of PMCA, end-point dilution titrations detected a > 320-fold increase in infectivity versus that in controls. These results provide strong support for the hypothesis that the agent of prion infectivity is not a virus. PMCA-generated samples caused the same clinical disease and neuropathology with the same rapid incubation period as the input brain-derived scrapie samples, providing no evidence for generation of a new strain in PMCA. However, the ratio of the infectivity titer to the amount of PrPres (specific infectivity) was much lower in PMCA versus brain-derived samples, suggesting the possibility that a substantial portion of PrPres generated in PMCA might be noninfectious.

引用

页码：E1244 / E1253

页数：10

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