Torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples
Abstract Background Torquetenovirus (TTV) belongs to Anelloviridae family, infects nearly all people indefinitely without causing overt disease establishing a fine and successful interaction with the host. Increasing evidence have shown some human viruses exploit extracellular vesicles thereby helpi...
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doaj-132199c033074aba94015cefbcbcb3382020-11-25T02:04:19ZengBMCVirology Journal1743-422X2018-09-0115111010.1186/s12985-018-1055-yTorquetenovirus detection in exosomes enriched vesicles circulating in human plasma samplesFrancesco Martelli0Lisa Macera1Pietro Giorgio Spezia2Chiara Medici3Mauro Pistello4Daniele Guasti5Paolo Romagnoli6Fabrizio Maggi7Simone Giannecchini8Department of Experimental and Clinical Medicine, University of FlorenceVirology Unit, Pisa University HospitalVirology Unit, Pisa University HospitalVirology Unit, Pisa University HospitalVirology Unit, Pisa University HospitalDepartment of Experimental and Clinical Medicine, University of FlorenceDepartment of Experimental and Clinical Medicine, University of FlorenceVirology Unit, Pisa University HospitalDepartment of Experimental and Clinical Medicine, University of FlorenceAbstract Background Torquetenovirus (TTV) belongs to Anelloviridae family, infects nearly all people indefinitely without causing overt disease establishing a fine and successful interaction with the host. Increasing evidence have shown some human viruses exploit extracellular vesicles thereby helping viral persistence in the host. Here, the presence of TTV in extracellular vesicles circulating in human plasma was investigated. Methods TTV DNA was quantified in plasma-derived exosomes from 122 samples collected from 97 diseased patients and 25 healthy donors. Exosomes enriched vesicles (EEVs) were extracted from plasma and characterized by Nanoparticle tracking analysis, by western blot for presence of tetraspanin CD63, CD81 and annexin II protein and, finally, by electron microscopy (EM). Presence and quantitation of TTV DNA were assessed with an universal single step real-time TaqMan PCR assay. Results Preliminary investigation showed that the human plasma extracted extracellular vesicles exhibited a main size of 70 nm, had concentration of 2.5 × 109/ml, and scored positive for tetraspanin CD63, CD81 and annexin II, typical characteristic of the exosomes vesicles. EEVs extracted from pooled plasma with TTV DNA viremia of 9.7 × 104 copies/ml showed to contain 6.3 × 102 TTV copies/ml, corresponding to 0.65% of total viral load. Important, TTV yield changed significantly following freezing/thawing, detergents and DNAse treatment of plasma before EEVs extraction. EEVs purified by sucrose-density gradient centrifugation and analysis of gradient fraction positive for exosomes marker CD63 harbored 102 TTV copies/ml. Moreover, EM evidenced the presence of TTV-like particles in EEVs. Successive investigation of plasma EEVs from 122 subjects (37 HIV-positive, 20 HCV infected, 20 HBV infected, 20 kidney transplant recipients, and 25 healthy) reported TTV DNA detection in 42 (34%) of the viremic samples (37 were from diseased patients and 5 from healthy people) at a mean level of 4.8 × 103 copies/ml. The examination of EEVs selected samples reported the presence of TTV genogroup 1, 3, 4 and 5, with genogroup 3 highly observed. Conclusions Collectively, although these observations should be confirmed by further studies, circulation of TTV particles in EEVs opens new avenues and mechanistic insights on the molecular strategies adopted by anelloviruses to persist in the host.http://link.springer.com/article/10.1186/s12985-018-1055-yAnellovirusesTorquetenovirusExosomesDNA viral loadViral persistenceHIV, transplant recipients |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Francesco Martelli Lisa Macera Pietro Giorgio Spezia Chiara Medici Mauro Pistello Daniele Guasti Paolo Romagnoli Fabrizio Maggi Simone Giannecchini |
spellingShingle |
Francesco Martelli Lisa Macera Pietro Giorgio Spezia Chiara Medici Mauro Pistello Daniele Guasti Paolo Romagnoli Fabrizio Maggi Simone Giannecchini Torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples Virology Journal Anelloviruses Torquetenovirus Exosomes DNA viral load Viral persistence HIV, transplant recipients |
author_facet |
Francesco Martelli Lisa Macera Pietro Giorgio Spezia Chiara Medici Mauro Pistello Daniele Guasti Paolo Romagnoli Fabrizio Maggi Simone Giannecchini |
author_sort |
Francesco Martelli |
title |
Torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples |
title_short |
Torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples |
title_full |
Torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples |
title_fullStr |
Torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples |
title_full_unstemmed |
Torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples |
title_sort |
torquetenovirus detection in exosomes enriched vesicles circulating in human plasma samples |
publisher |
BMC |
series |
Virology Journal |
issn |
1743-422X |
publishDate |
2018-09-01 |
description |
Abstract Background Torquetenovirus (TTV) belongs to Anelloviridae family, infects nearly all people indefinitely without causing overt disease establishing a fine and successful interaction with the host. Increasing evidence have shown some human viruses exploit extracellular vesicles thereby helping viral persistence in the host. Here, the presence of TTV in extracellular vesicles circulating in human plasma was investigated. Methods TTV DNA was quantified in plasma-derived exosomes from 122 samples collected from 97 diseased patients and 25 healthy donors. Exosomes enriched vesicles (EEVs) were extracted from plasma and characterized by Nanoparticle tracking analysis, by western blot for presence of tetraspanin CD63, CD81 and annexin II protein and, finally, by electron microscopy (EM). Presence and quantitation of TTV DNA were assessed with an universal single step real-time TaqMan PCR assay. Results Preliminary investigation showed that the human plasma extracted extracellular vesicles exhibited a main size of 70 nm, had concentration of 2.5 × 109/ml, and scored positive for tetraspanin CD63, CD81 and annexin II, typical characteristic of the exosomes vesicles. EEVs extracted from pooled plasma with TTV DNA viremia of 9.7 × 104 copies/ml showed to contain 6.3 × 102 TTV copies/ml, corresponding to 0.65% of total viral load. Important, TTV yield changed significantly following freezing/thawing, detergents and DNAse treatment of plasma before EEVs extraction. EEVs purified by sucrose-density gradient centrifugation and analysis of gradient fraction positive for exosomes marker CD63 harbored 102 TTV copies/ml. Moreover, EM evidenced the presence of TTV-like particles in EEVs. Successive investigation of plasma EEVs from 122 subjects (37 HIV-positive, 20 HCV infected, 20 HBV infected, 20 kidney transplant recipients, and 25 healthy) reported TTV DNA detection in 42 (34%) of the viremic samples (37 were from diseased patients and 5 from healthy people) at a mean level of 4.8 × 103 copies/ml. The examination of EEVs selected samples reported the presence of TTV genogroup 1, 3, 4 and 5, with genogroup 3 highly observed. Conclusions Collectively, although these observations should be confirmed by further studies, circulation of TTV particles in EEVs opens new avenues and mechanistic insights on the molecular strategies adopted by anelloviruses to persist in the host. |
topic |
Anelloviruses Torquetenovirus Exosomes DNA viral load Viral persistence HIV, transplant recipients |
url |
http://link.springer.com/article/10.1186/s12985-018-1055-y |
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