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Updated: Jun 28, 2026
Protein Misfolding Cyclic Amplification of Prions
Published on: November 7, 2012
In vitro strain adaptation of CWD prions by serial protein misfolding cyclic amplification
Crystal Meyerett1, Brady Michel, Bruce Pulford
1Department of Microbiology, Immunology, and Pathology, Colorado State University, College of Veterinary Medicine and Biomedical Sciences, Fort Collins, CO 80523, USA.
Abstract:
We used serial protein misfolding cyclic amplification (sPMCA) to amplify the D10 strain of CWD prions in a linear relationship over two logs of D10 dilutions. The resultant PMCA-amplified D10 induced terminal TSE disease in CWD-susceptible Tg(cerPrP)1536 mice with a survival time approximately 80 days shorter than the original D10 inoculum, similar to that produced by in vivo sub-passage of D10 in Tg(cerPrP)1536 mice. Both in vitro-amplified and mouse-passaged D10 produced brain lesion profiles, glycoform ratios and conformational stabilities significantly different than those produced by the original D10 inoculum in Tg(cerPrP)1536 mice. These findings demonstrate that sPMCA can amplify and adapt prion strains in vitro as effectively and much more quickly than in vivo strain adaptation by mouse passage. Thus sPMCA may represent a powerful tool to assess prion strain adaptation and species barriers in vitro.
Insights
Serial protein misfolding cyclic amplification (sPMCA) effectively amplifies and adapts CWD prion strains in vitro. This method is faster than traditional mouse passage for studying prion adaptation and species barriers.
Area of Science:
- Neuroscience
- Biochemistry
- Molecular Biology
Background:
- Chronic Wasting Disease (CWD) is a transmissible spongiform encephalopathy (TSE) affecting cervids.
- Prion strain adaptation is crucial for understanding disease transmission and developing diagnostics.
- In vivo methods for prion strain adaptation are time-consuming.
Purpose of the Study:
- To evaluate the efficacy of serial protein misfolding cyclic amplification (sPMCA) for amplifying and adapting CWD prion strains.
- To compare in vitro sPMCA adaptation with in vivo mouse passage adaptation.
Main Methods:
- Serial protein misfolding cyclic amplification (sPMCA) was used to amplify the D10 strain of CWD prions.
- The amplified prions were inoculated into CWD-susceptible Tg(cerPrP)1536 mice.
- Brain lesion profiles, glycoform ratios, and conformational stabilities were analyzed.
Main Results:
- sPMCA linearly amplified CWD prions (D10 strain) over a two-log dilution range.
- PMCA-amplified D10 induced terminal TSE disease in mice with a significantly shorter survival time compared to the original inoculum.
- Both in vitro-amplified and mouse-passaged D10 exhibited distinct brain lesion profiles, glycoform ratios, and conformational stabilities compared to the original D10.
Conclusions:
- sPMCA effectively amplifies and adapts prion strains in vitro.
- sPMCA is a faster and equally effective method for prion strain adaptation compared to in vivo mouse passage.
- sPMCA is a powerful tool for assessing prion strain adaptation and species barriers in vitro.
