Human parvovirus B19 (B19V) is a single-stranded DNA virus. The genome encodes a multifunctional non-structural protein (NS), two capsid proteins (VP1, VP2) and other small non-structural proteins (7.5. kDa, 9. kDa, 11. kDa). Within sensitive cells, B19V can achieve a productive replicative cycle or, on the contrary, establish persistence; differences in its expression profile have been yet investigated following i. n vitro infections by methodologies enabling information on the entire infected cell population. Conversely, the present study reports quantitative data on the production of B19V analytes (DNA, RNAs, and proteins) at single cell-level, underlining cell-to-cell differences through the viral specific macromolecular synthesis process. Microscope imaging assays (in situ hybridization and immunocytochemical assays), exploiting chemiluminescence as principle detection and targeting viral nucleic acids and antigens, have been performed on a permissive cell line following in vitro infection. Chemiluminescence, involving the emission of photons deriving from a chemical reaction, provided the localization and quantitative detection of analytes down to a few molecules within infected cells. In our experimental conditions, B19V transcriptional activity, leading to the production of NS and VP RNAs, has been detected early in the viral cycle (from 12. h post-infection, hpi) and before genome replication, starting at 24. hpi. The analysis of VP RNAs and related proteins suggested an inhibitory effect on capsid protein translation, as a post-transcriptional regulation events. Indeed, high levels of VP transcripts have been detected at early stages of infection while the proteins accumulated within cells only at 48-72. hpi.

Bonvicini F, Mirasoli M, Manaresi E, Bua G, Calabria D, Roda A, et al. (2013). Single-cell chemiluminescence imaging of parvovirus B19 life cycle. VIRUS RESEARCH, 26, 517-521 [10.1016/j.virusres.2013.09.037].

Single-cell chemiluminescence imaging of parvovirus B19 life cycle

BONVICINI, FRANCESCA;MIRASOLI, MARA;MANARESI, ELISABETTA;BUA, GLORIA;CALABRIA, DONATO;RODA, ALDO;GALLINELLA, GIORGIO
2013

Abstract

Human parvovirus B19 (B19V) is a single-stranded DNA virus. The genome encodes a multifunctional non-structural protein (NS), two capsid proteins (VP1, VP2) and other small non-structural proteins (7.5. kDa, 9. kDa, 11. kDa). Within sensitive cells, B19V can achieve a productive replicative cycle or, on the contrary, establish persistence; differences in its expression profile have been yet investigated following i. n vitro infections by methodologies enabling information on the entire infected cell population. Conversely, the present study reports quantitative data on the production of B19V analytes (DNA, RNAs, and proteins) at single cell-level, underlining cell-to-cell differences through the viral specific macromolecular synthesis process. Microscope imaging assays (in situ hybridization and immunocytochemical assays), exploiting chemiluminescence as principle detection and targeting viral nucleic acids and antigens, have been performed on a permissive cell line following in vitro infection. Chemiluminescence, involving the emission of photons deriving from a chemical reaction, provided the localization and quantitative detection of analytes down to a few molecules within infected cells. In our experimental conditions, B19V transcriptional activity, leading to the production of NS and VP RNAs, has been detected early in the viral cycle (from 12. h post-infection, hpi) and before genome replication, starting at 24. hpi. The analysis of VP RNAs and related proteins suggested an inhibitory effect on capsid protein translation, as a post-transcriptional regulation events. Indeed, high levels of VP transcripts have been detected at early stages of infection while the proteins accumulated within cells only at 48-72. hpi.
2013
Bonvicini F, Mirasoli M, Manaresi E, Bua G, Calabria D, Roda A, et al. (2013). Single-cell chemiluminescence imaging of parvovirus B19 life cycle. VIRUS RESEARCH, 26, 517-521 [10.1016/j.virusres.2013.09.037].
Bonvicini F; Mirasoli M; Manaresi E; Bua G; Calabria D; Roda A; Gallinella G
File in questo prodotto:
Eventuali allegati, non sono esposti

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/205015
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo

Citazioni
  • ???jsp.display-item.citation.pmc??? 1
  • Scopus 8
  • ???jsp.display-item.citation.isi??? 7
social impact