Jorge moreno san vidal

We have conducted a series of experiments using the Protein Misfolding Cyclic Amplification PMCA technique that mimics in vitro some of the fundamental steps involved in prion replication in vivo, albeit with accelerated kinetics. The in vitro generated prions possess key prion features, i. We have used PMCA to efficiently replicate a variety of prion strains from, among others, mice, hamsters, bank voles, deer, cattle, sheep, and humans. The correlation between in vivo data and our in vitro results suggest that PMCA is a valuable tool for assessing the strength of the transmission barriers between diverse species and for different prion strains; we are using the method to determine which amino acids in the PrPC sequence contribute to the strength of the transmission barrier.

These studies are proving very useful in evaluating the potential risks to humans and animals, of not only established prion strains, but also new atypical strains. For example, while classical sheep scrapie is unable to cross the human transmission barrier in vitro, bovine spongiform encephalopathy BSE propagated in sheep does so efficiently.

In addition, we have also generated prions that are infectious to species hitherto considered to be resistant to prion disease. Understanding the key features of the spontaneous formation of bona fide prions through a novel methodology that enables their swift and consistent generation. An optimized Western blot assay provides a comprehensive assessment of the physiological endoproteolytic processing of the prion protein.