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Prion early kinetics revisited using a streptomycin-based PrP(res) extraction method
Edwige Leclere1, Aly Moussa, Latifa Chouaf-Lakhdar
1Laboratoire de Virologie Médicale, Centre Hospitalier Universitaire A. Michalon, BP17X 38043 Grenoble, France.
Abstract:
The use of streptomycin in the PrP(sc) detection procedures represents a new and attractive way to detect more PrP(sc), the best marker for the transmissible spongiform encephalopathies (TSEs). Actually, the streptomycin PrP(sc) aggregating property reported recently was established as beneficial in PrP(sc) detection using immunohistochemistry in diagnostic as well as in experimental conditions. The present study reports in details how to use advantageously this original streptomycin property in PrP(res) biochemical extraction and detection. Using TSE diagnostic brain material, specificity and increased sensitivity using streptomycin-treated samples were substantiated. Then an early sequential brain and spleen sampling (from 7 to 49 days post-inoculation) from C57Bl/6 mice inoculated intra-cerebrally or intra-peritoneally with C506M3 scrapie strain was analysed using streptomycin versus ultracentrifugation PrP(res) extraction. Whatever the inoculation route, streptomycin allowed earlier PrP(res) detection in spleen (7 d.p.i.), then in brain suggesting a stronger affinity of the infectious agent for the lymphoid compartment.
Insights
Streptomycin enhances transmissible spongiform encephalopathy (TSE) detection by improving PrP(sc) aggregation. This method allows earlier detection of PrP(res) in spleen and brain tissues, aiding in diagnostic and experimental research.
Area of Science:
- Neuroscience
- Biochemistry
- Pathology
Background:
- Transmissible spongiform encephalopathies (TSEs) are characterized by the accumulation of misfolded prion protein (PrP(sc)).
- Current detection methods for PrP(sc) can be limited in sensitivity and scope.
- Streptomycin has recently shown potential in enhancing PrP(sc) detection through aggregation properties.
Purpose of the Study:
- To detail the methodology for utilizing streptomycin's PrP(sc) aggregating property in PrP(res) biochemical extraction and detection.
- To evaluate the specificity and sensitivity of streptomycin-assisted PrP(res) detection using TSE diagnostic brain material.
- To assess the efficacy of streptomycin for early PrP(res) detection in various tissues and inoculation routes in a mouse model.
Main Methods:
- Streptomycin-based PrP(res) extraction and detection protocols were developed and detailed.
- TSE diagnostic brain material was analyzed to compare streptomycin-treated samples with conventional methods.
- C57Bl/6 mice inoculated with scrapie were sampled sequentially (7-49 days post-inoculation) for brain and spleen analysis, comparing streptomycin extraction with ultracentrifugation.
Main Results:
- Streptomycin treatment substantiated increased sensitivity and specificity in PrP(res) detection in TSE diagnostic samples.
- Early PrP(res) detection in spleen (as early as 7 days post-inoculation) was achieved using streptomycin, irrespective of the inoculation route.
- Earlier detection of PrP(res) in the brain was also observed with streptomycin treatment, following spleen detection.
Conclusions:
- Streptomycin offers a novel and advantageous method for PrP(res) extraction and detection, enhancing sensitivity and specificity.
- The use of streptomycin facilitates earlier diagnosis of TSEs by enabling earlier detection of PrP(res) in affected tissues.
- The findings suggest a preferential affinity of the infectious agent for the lymphoid compartment, as indicated by earlier PrP(res) detection in the spleen.
