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[Antiviral effect of antisense oligodeoxynucleotides complementary to hepatitis B virus X gene in vitro]
1Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an.
Abstract:
It has been reported that the products of the human hepatitis B virus (HBV) X genes can transactivate a variety of viral and host promoters including the X, core, pre S2/S and pre S1 promoter. In order to investigate their antiviral effect in cell culture, three pieces of antisense phosphorothioate oligodeoxynucleotides (ASON) complementary to HBV X genes 1510-1530, 1555-1581, 1768-1791 regions were synthesized. The specificity of the inhibitory effect of the ASON was determined by using 2, 2, 15 cells by ELISA, PAP-ELISA and in situ hybridization to detect HBsAg, HBeAg, HBxAg and HBV DNA. The results showed that these ASON could inhibit the expression of HBxAg, as well as HBsAg and HBeAg, with the inhibitary rates of 78.07%, 80.65% and 62.76% respectively. In situ hybridization results indicated that HBV DNA replication can also be inhibited. The decrease in virus production and the ammount of HBV DNA were dose and time dependent. The mechanism of action may be due to the inhibition of HBxAg by sequences specific to ASON, and then the decrease of its transactivating effect for HBV DNA promoter.
Insights
Antisense oligodeoxynucleotides (ASON) targeting hepatitis B virus (HBV) X genes show significant antiviral effects. These ASON inhibit HBV replication and the expression of key viral proteins, offering a potential therapeutic strategy.
Area of Science:
- Virology
- Molecular Biology
- Antiviral Therapeutics
Background:
- Hepatitis B virus (HBV) X gene products transactivate viral and host promoters.
- Understanding HBV X gene function is crucial for developing antiviral strategies.
Purpose of the Study:
- To investigate the antiviral effect of antisense phosphorothioate oligodeoxynucleotides (ASON) against HBV X genes in cell culture.
- To determine the specificity and efficacy of ASON in inhibiting HBV replication and protein expression.
Main Methods:
- Synthesis of three ASON targeting specific regions of the HBV X gene.
- Detection of HBsAg, HBeAg, HBxAg, and HBV DNA using ELISA, PAP-ELISA, and in situ hybridization.
- Assessment of dose- and time-dependent effects of ASON.
Main Results:
- ASON demonstrated significant inhibition of HBxAg (78.07%), HBsAg (80.65%), and HBeAg (62.76%) expression.
- In situ hybridization confirmed inhibition of HBV DNA replication.
- Virus production and HBV DNA levels decreased in a dose- and time-dependent manner.
Conclusions:
- ASON effectively inhibit HBV X gene expression and its transactivating effects.
- These findings suggest ASON as a promising approach for inhibiting HBV replication and viral protein production.