BK Polyomavirus MicroRNA Levels in Exosomes Are Modulated by Non-Coding Control Region Activity and Down-Regulate Viral Replication When Delivered to Non-Infected Cells Prior to Infection
Abstract
:1. Introduction
2. Materials and Methods
2.1. Urine Sampling
2.2. Cell Cultures and BkPyV Molecular Clones
2.3. BKPyV Infection of Cos-7 and RPTECs
2.4. Plasmid Exosomes BKPyV miRNA Expression in Cos-7 Cells
2.5. Exosome Enriched Vesicles Extraction
2.6. Exosome Addition and Anti BKPyV-miRNA Inhibition in Cos-7 Cells
2.7. BKPyV DNA Quantification
2.8. BKPyV Pre-miRNA and Mature miRNA Quantification
2.9. siRNA knock-Down of Sp1 and Immunoblotting
2.10. NCCR Sequencing
2.11. Statistical Tests
3. Results
3.1. Rearranged NCCR-BKPyV Variants Show Higher Viral Loads But 10-Fold Lower miRNA Expression than Archetype NCCR BKPyV
3.2. Sp1 Expression Levels Modulate BKPyV miRNA Levels in Archetype NCCR Virus And Variants With Low EVGR Expression But Not in Viral Variants Having an Activated EVGR Expression
3.3. BKPyV miRNA Levels in Primary Human Renal Tubular Epithelial Cells (RPTECs) Inversely Correlate with the Evgr-Activity and Replication Rate of Archetype and rr-NCCR-Viruses
3.4. Pre-Infection Addition of BKPyV-Exosome Preparations Containing BKPyV miRNA Inhibit BKPyV Replication of Dunlop or sp1-4 Mutant But Have No Effect On Archetype BKPyV or sp1-2 Mutant Strains
3.5. Analysis of the BKPyV NCCR Architecture and Exosomes Content in Urine Samples of Immunocompromised Patients
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Martelli, F.; Wu, Z.; Delbue, S.; Weissbach, F.H.; Giulioli, M.C.; Ferrante, P.; Hirsch, H.H.; Giannecchini, S. BK Polyomavirus MicroRNA Levels in Exosomes Are Modulated by Non-Coding Control Region Activity and Down-Regulate Viral Replication When Delivered to Non-Infected Cells Prior to Infection. Viruses 2018, 10, 466. https://doi.org/10.3390/v10090466
Martelli F, Wu Z, Delbue S, Weissbach FH, Giulioli MC, Ferrante P, Hirsch HH, Giannecchini S. BK Polyomavirus MicroRNA Levels in Exosomes Are Modulated by Non-Coding Control Region Activity and Down-Regulate Viral Replication When Delivered to Non-Infected Cells Prior to Infection. Viruses. 2018; 10(9):466. https://doi.org/10.3390/v10090466
Chicago/Turabian StyleMartelli, Francesco, Zongsong Wu, Serena Delbue, Fabian H. Weissbach, Maria Chiara Giulioli, Pasquale Ferrante, Hans H. Hirsch, and Simone Giannecchini. 2018. "BK Polyomavirus MicroRNA Levels in Exosomes Are Modulated by Non-Coding Control Region Activity and Down-Regulate Viral Replication When Delivered to Non-Infected Cells Prior to Infection" Viruses 10, no. 9: 466. https://doi.org/10.3390/v10090466
APA StyleMartelli, F., Wu, Z., Delbue, S., Weissbach, F. H., Giulioli, M. C., Ferrante, P., Hirsch, H. H., & Giannecchini, S. (2018). BK Polyomavirus MicroRNA Levels in Exosomes Are Modulated by Non-Coding Control Region Activity and Down-Regulate Viral Replication When Delivered to Non-Infected Cells Prior to Infection. Viruses, 10(9), 466. https://doi.org/10.3390/v10090466