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Substitution rtq267h of hepatitis B virus increases the weight of replication and Lamivudine resistance.

Qin B, Zhang B, Zhang X, He T, Xu W, Fu L, Tu C - Hepat Mon (2013)

Bottom Line: The in vitro susceptibility analysis showed that the existence of rtQ267H in WT and LMV-resistant (LMVr) HBV were responsible for the reduced susceptibility to LMV to varying degrees, and enhanced HBV replication capacity.However, HBV harbored this substitution retained normal susceptibility to ADV, LdT, ETV, and TDF.The result suggested that rtQ267H is a potential adaptive mutation of HBV to LMV.

View Article: PubMed Central - PubMed

Affiliation: Shaoxing Centre for Disease Control and Prevention, Shaoxing, China ; State Key Lab of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.

ABSTRACT

Background: Nucleus(t)ide analogs (NAs), containing Lamivudine (LMV), adefovir dipivoxil (ADV), endeavor (ETV), telbivudine (LdT), and tenofovir (TDF) are widely used for the treatment of chronic hepatitis B (CHB), but long term anti-Hepatitis B virus (HBV) therapy with NAs may give rise to the emergence of drug-resistant viral mutants.

Objectives: This study aimed to find and identify some new resistance mutations of HBV from the patients accepted anti-HBV therapy.

Patients and methods: The reverse transcriptase (RT) coding region of HBV was PCR-amplified using HBV DNA extracted from patients' blood samples and sequenced.

Results: Nineteen substitution mutations were detected. Among them, rtQ267H was often observed in patients receiving LMV administration. This LMV therapy-related mutation was introduced into HBV replication-competent plasmids. The in vitro susceptibility of both wild-type (WT) and mutant-type (MT) HBV to NAs was analyzed by Southern blot, and/or quantitative real-time PCR (qRT-PCR). The rtQ267H substitution enhanced HBV replication not merely in single-site mutation, but also in multisite mutations. The in vitro susceptibility analysis showed that the existence of rtQ267H in WT and LMV-resistant (LMVr) HBV were responsible for the reduced susceptibility to LMV to varying degrees, and enhanced HBV replication capacity. However, HBV harbored this substitution retained normal susceptibility to ADV, LdT, ETV, and TDF.

Conclusions: The result suggested that rtQ267H is a potential adaptive mutation of HBV to LMV.

No MeSH data available.


Related in: MedlinePlus

Southern Blot Analysis to Detect the Anti-HBV Effect of LMV towards WT HBV and Different MutantsHuh7 cells were transfected with pHBV1.3 (Figure 3 A), -rtQ267H (Figure 3 B), -rtM204V/Q267H (Figure 3 C), -rtL180M/M204V (Figure 3 D), -rtL180M/M204V/Q267H (Figure 3 E), and then treated with LMV at the indicated concentrations. Encapsulated HBV DNA was extracted and analyzed by Southern blot (upper panel). The relative level of LMV’s anti-HBV effect is shown as a percentage of the control by gray analysis (lower panel).
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fig6188: Southern Blot Analysis to Detect the Anti-HBV Effect of LMV towards WT HBV and Different MutantsHuh7 cells were transfected with pHBV1.3 (Figure 3 A), -rtQ267H (Figure 3 B), -rtM204V/Q267H (Figure 3 C), -rtL180M/M204V (Figure 3 D), -rtL180M/M204V/Q267H (Figure 3 E), and then treated with LMV at the indicated concentrations. Encapsulated HBV DNA was extracted and analyzed by Southern blot (upper panel). The relative level of LMV’s anti-HBV effect is shown as a percentage of the control by gray analysis (lower panel).

Mentions: To determine whether rtQ267H influenced HBV susceptibility to LMV and others NAs, Huh7 cells were transfected with MT or WT HBV plasmids, then exposed to the indicated NA concentrations and the HBV core-associated DNA was analyzed by Southern blot. As shown in Figure 3 A, LMV inhibited the replication of WT HBV in a dose-dependent manner, while the susceptibility of pHBV1.3-rtQ267H, -rtM204V/Q267H, -rtL180M/M204V and -rtL180M/M204V/Q267H to LMV and LdT was greatly reduced


Substitution rtq267h of hepatitis B virus increases the weight of replication and Lamivudine resistance.

Qin B, Zhang B, Zhang X, He T, Xu W, Fu L, Tu C - Hepat Mon (2013)

Southern Blot Analysis to Detect the Anti-HBV Effect of LMV towards WT HBV and Different MutantsHuh7 cells were transfected with pHBV1.3 (Figure 3 A), -rtQ267H (Figure 3 B), -rtM204V/Q267H (Figure 3 C), -rtL180M/M204V (Figure 3 D), -rtL180M/M204V/Q267H (Figure 3 E), and then treated with LMV at the indicated concentrations. Encapsulated HBV DNA was extracted and analyzed by Southern blot (upper panel). The relative level of LMV’s anti-HBV effect is shown as a percentage of the control by gray analysis (lower panel).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3842524&req=5

fig6188: Southern Blot Analysis to Detect the Anti-HBV Effect of LMV towards WT HBV and Different MutantsHuh7 cells were transfected with pHBV1.3 (Figure 3 A), -rtQ267H (Figure 3 B), -rtM204V/Q267H (Figure 3 C), -rtL180M/M204V (Figure 3 D), -rtL180M/M204V/Q267H (Figure 3 E), and then treated with LMV at the indicated concentrations. Encapsulated HBV DNA was extracted and analyzed by Southern blot (upper panel). The relative level of LMV’s anti-HBV effect is shown as a percentage of the control by gray analysis (lower panel).
Mentions: To determine whether rtQ267H influenced HBV susceptibility to LMV and others NAs, Huh7 cells were transfected with MT or WT HBV plasmids, then exposed to the indicated NA concentrations and the HBV core-associated DNA was analyzed by Southern blot. As shown in Figure 3 A, LMV inhibited the replication of WT HBV in a dose-dependent manner, while the susceptibility of pHBV1.3-rtQ267H, -rtM204V/Q267H, -rtL180M/M204V and -rtL180M/M204V/Q267H to LMV and LdT was greatly reduced

Bottom Line: The in vitro susceptibility analysis showed that the existence of rtQ267H in WT and LMV-resistant (LMVr) HBV were responsible for the reduced susceptibility to LMV to varying degrees, and enhanced HBV replication capacity.However, HBV harbored this substitution retained normal susceptibility to ADV, LdT, ETV, and TDF.The result suggested that rtQ267H is a potential adaptive mutation of HBV to LMV.

View Article: PubMed Central - PubMed

Affiliation: Shaoxing Centre for Disease Control and Prevention, Shaoxing, China ; State Key Lab of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.

ABSTRACT

Background: Nucleus(t)ide analogs (NAs), containing Lamivudine (LMV), adefovir dipivoxil (ADV), endeavor (ETV), telbivudine (LdT), and tenofovir (TDF) are widely used for the treatment of chronic hepatitis B (CHB), but long term anti-Hepatitis B virus (HBV) therapy with NAs may give rise to the emergence of drug-resistant viral mutants.

Objectives: This study aimed to find and identify some new resistance mutations of HBV from the patients accepted anti-HBV therapy.

Patients and methods: The reverse transcriptase (RT) coding region of HBV was PCR-amplified using HBV DNA extracted from patients' blood samples and sequenced.

Results: Nineteen substitution mutations were detected. Among them, rtQ267H was often observed in patients receiving LMV administration. This LMV therapy-related mutation was introduced into HBV replication-competent plasmids. The in vitro susceptibility of both wild-type (WT) and mutant-type (MT) HBV to NAs was analyzed by Southern blot, and/or quantitative real-time PCR (qRT-PCR). The rtQ267H substitution enhanced HBV replication not merely in single-site mutation, but also in multisite mutations. The in vitro susceptibility analysis showed that the existence of rtQ267H in WT and LMV-resistant (LMVr) HBV were responsible for the reduced susceptibility to LMV to varying degrees, and enhanced HBV replication capacity. However, HBV harbored this substitution retained normal susceptibility to ADV, LdT, ETV, and TDF.

Conclusions: The result suggested that rtQ267H is a potential adaptive mutation of HBV to LMV.

No MeSH data available.


Related in: MedlinePlus