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Inhibition of rabies virus multiplication by siRNA delivered through adenoviral vector in vitro in BHK-21 cells and in vivo in mice.
- Source :
-
Research in veterinary science [Res Vet Sci] 2012 Aug; Vol. 93 (1), pp. 498-503. Date of Electronic Publication: 2011 Jul 20. - Publication Year :
- 2012
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Abstract
- To evaluate antiviral potential of adenoviral vector-delivered small interfering RNA (siRNA) against rabies, recombinant, replication-defective adenoviral vectors (rAdV) encoding siRNAs targeting rabies virus (RV) polymerase (L) and nucleoprotein (N) genes were developed. The siRNAs were delivered as small hairpin RNAs (shRNAs) through these vectors. Treatment of BHK-21 cells with rAdV expressing siRNA targeting L gene (rAdV-L) and N gene (rAdV-N) (100 MOI) and their subsequent infection with RV (0.001 MOI, RV PV-11), reduced RV fluorescent foci by 48.2% (mean±SEM; 48.17±0.6540, N=6) and 41.8% (mean±SEM; 41.83±0.3073, N=6), respectively, with respect to that of BHK-21 cells treated with rAdV expressing negative control siRNA (rAdV-Neg) indicating inhibition of multiplication of RV in BHK-21 cells in response to adenoviral vector mediated siRNA delivery. Also, the similar treatment of BHK-21 cells with rAdV-L and rAdV-N and similar subsequent infection of them with RV resulted in reduction in RV mRNA transcript levels for their respective targets (RV L gene for rAdV-L and N gene for rAdV-N). mRNA transcript level for RV L gene was reduced by 17.88-fold (mean±SEM; 17.88±0.06638, N=6) in cells treated with rAdV-L and that for RV N gene was reduced by 5.7-fold (mean±SEM; 5.7±0.04472, N=6), in cells treated with rAdV-N, in comparison with that in cells treated with rAdV-Neg, as analyzed by using real-time PCR. These in vitro studies showed that between these two, adenoviral vector mediated delivery of siRNA targeting RV L gene was comparatively more effective in inhibiting RV multiplication in BHK-21 cells than that of siRNA targeting RV N gene (p<0.0001). Localized treatment (intramuscular injection in masseter muscle) of mice with 10(7) plaque forming units of either rAdV-L or rAdV-N and subsequent lethal RV infection (15-20LD(50) of CVS-11) at the same site, through the same route, although resulted in 50% protection (3 out of 6 mice survived) against lethal rabies, the survival patterns for groups of mice treated with either rAdV-L or rAdV-N and that treated with rAdV-Neg did not differ significantly (p=0.5234). These results indicated that adenoviral vector mediated siRNA delivery, in vitro in BHK-21 cells inhibited RV multiplication in vitro in BHK-21 cells; siRNA targeting RV L gene used in this study was comparatively more efficient in doing this than that targeting RV N gene used in this study; in vivo in mice inhibited RV multiplication in mice and imparted partial protection against lethal rabies and so it may have a potential to be used as an alternative antiviral approach against rabies, although further study is required to establish its efficacy for this purpose.<br /> (Copyright © 2011 Elsevier Ltd. All rights reserved.)
- Subjects :
- Adenoviridae genetics
Animals
Cell Line
Cricetinae
Female
Genetic Vectors genetics
Humans
In Vitro Techniques
Male
Mice
RNA, Small Interfering genetics
Rabies virus genetics
Rabies virus physiology
Virus Replication drug effects
Antiviral Agents therapeutic use
RNA, Small Interfering therapeutic use
Rabies drug therapy
Rabies virus drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 1532-2661
- Volume :
- 93
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Research in veterinary science
- Publication Type :
- Academic Journal
- Accession number :
- 21767857
- Full Text :
- https://doi.org/10.1016/j.rvsc.2011.06.008