Rapid communicationInhibition of hepatitis B virus replication in vivo by nucleoside analogues and siRNA☆
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DNA constructs and siRNA
An HBV replication competent vector (pHBV 1.5) was used for all experiments as described earlier.1, 12 Twenty-one nucleotide siRNA was designed and purchased from Dharmacon Research Inc. in the ready-to-use option. Two different locations in the HBV genome were selected, located either in the coding region of the S-gene or the core (C) gene. The nucleotide mRNA target sequence in the S-gene is the following: 5′AAG CCU UAG AGU CUC CUG AGC-3′ (207-227) and in the core gene: 5′A AUU UGU UCA GUC
High-volume injection of a HBV replication competent vector results in serum and liver expression of HBV proteins
Earlier experiments indicated that high-volume injection of plasmid DNA results in liver transgene expression.16 We thus performed high-volume injection via tail vein of 20 μg of an HBV replication competent vector (pHBV 1.5) in mice and studied HBsAg and HBeAg serum expression. At least 3 animals were treated in parallel per time point indicated. Three and 4 days after pHBV 1.5 injection, maximal serum levels for HBsAg and HBeAg were found, respectively. At later time points, serum expression
Discussion
In the present study, we have developed a novel mouse model to study HBV replication in vivo. After transfer of an HBV replication competent vectors through high-volume tail injection, more than 10% of mouse hepatocytes became positive for HBV protein expression. Interestingly, further analysis indicated that viral capsids were assembled and began replicating the viral genome and were secreted into the serum of the animals as virions; thus, most components of the HBV life cycle can now be
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Cited by (171)
Treatment of Hepatitis B
2017, Zakim and Boyer's Hepatology: A Textbook of Liver DiseaseRecent developments in antivirals against hepatitis B virus
2016, Virus ResearchNew therapeutic agents for chronic hepatitis B
2016, The Lancet Infectious DiseasesTargeting the Achilles heel of the hepatitis B virus: A review of current treatments against covalently closed circular DNA
2015, Drug Discovery TodayCitation Excerpt :Previous studies have shown that siRNA (21–25 nucleotides) directed against the core and/or HBx transcripts can inhibit HBV gene expression in HBV-infected cells and significantly decrease the levels of HBsAg, HBeAg, HBV RNA and HBV DNA [77]. Other studies also have highlighted potential synergism between siRNA and other anti-HBV treatment strategies, such as NAs [78]. Another recent and interesting study, by Li et al. [79], showed that a siRNA targeting the NLS of the HBcAg core protein can inhibit cccDNA recycling (intracellular amplification) [79].
Mouse models of hepatitis B and delta virus infection
2014, Journal of Immunological MethodsCitation Excerpt :However, HBV infection persisted for at least 81 days after hydrodynamic injection of mice lacking adaptive immune cells and natural killer cells, thus demonstrating that the outcome of hydrodynamic transfection of HBV depended on the host immune response (Yang et al., 2002). Hydrodynamic injection studies also showed that simultaneous delivery of HBV expressing plasmids and HBV-specific siRNAs prevented HBV replication (Klein et al., 2003; McCaffrey et al., 2003), while by injecting modified HBV DNA plasmids into C57BL/6 mice, a significant immune clearance of HBV could be achieved (Lin et al., 2010). By avoiding to raise sufficient cellular immunity against HBcAg, the authors proposed a model of HBV persistence, which better resembled the immune tolerant stage of chronic HBV carriers.
Hepatitis B Virus Research in South Africa
2022, Viruses
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Supported by a grant in the HepNet (to C.T. and M.P.M.) and the “fortüne-program” of the University of Tübingen, grant 1037-1-0 (to C.T.B.).
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C. Klein and C.T. Bock contributed equally to this work.