Lamivudine-resistant Hepatitis Research Articles

Virological responses to tenofovir-alafenamide-containing antiretroviral therapy in people living with HIV co-infected with lamivudine-resistant or lamivudine-susceptible hepatitis B virus

BackgroundData on the effectiveness of tenofovir alafenamide (TAF) against lamivudine-resistant (LAM-R) hepatitis B virus (HBV) among patients co-infected with human immunodeficiency virus (HIV) and HBV are limited. MethodsBetween April and December 2018, HIV-positive patients co-infected with LAM-R or lamivudine-susceptible (LAM-S) HBV who switched from tenofovir-disoproxil-fumarate-containing antiretroviral therapy (ART) to TAF-containing ART were followed for 96 weeks. Plasma HBV and HIV loads, HBV serological markers, and liver function before and after the switch were analysed. ResultsIn total, 182 patients co-infected with HIV and HBV were included in this study: 45 with LAM-R HBV and 137 with LAM-S HBV. At baseline, 28.9% and 7.4% of patients in the LAM-R and LAM-S groups, respectively, tested positive for hepatitis B virus envelope antigen (HBeAg) (P<0.001), and the respective percentages of patients who had achieved plasma HBV DNA <20 IU/mL were 95.5% and 97.1%. At weeks 48 and 96, 100% and 94.9% of patients in the LAM-R group, respectively, and 97.1% and 95.6% of patients in the LAM-S group, respectively, maintained plasma HBV DNA <20 IU/mL. Lamivudine resistance of HBV and baseline hepatitis B virus surface antigen (HBsAg) level were associated with HBsAg decrement at week 96 at a degree of 0.25 log10 IU/mL [95% confidence interval (CI) 0.059–0.246] and 0.22 log10 IU/mL (per 1-log10IU/mL increase, 95% CI 0.018–0.101), respectively. At week 96, 2.2% (4/182) of patients had HBsAg loss; no patients in the LAM-R group and 25.0% (2/8) of patients in the LAM-S group had HBeAg seroconversion. ConclusionsSwitching to TAF-containing regimens maintained high rates of HBV viral suppression in patients co-infected with either LAM-R or LAM-S HBV. The decrease in HBsAg was minimal, and HBsAg seroconversion occurred infrequently.

Continued lamivudine for adults with lamivudine-resistant chronic hepatitis B virus infection

Continued lamivudine for adults with lamivudine-resistant chronic hepatitis B virus infection

Emtricitabine for adults with lamivudine-resistant chronic hepatitis B virus infection

Emtricitabine for adults with lamivudine-resistant chronic hepatitis B virus infection

Novel fluoronucleoside analog NCC inhibits lamivudine-resistant hepatitis B virus in a hepatocyte model

Antiviral drug resistance is the most important factor contributing to treatment failure using nucleos(t)ide analogs such as lamivudine for chronic infection with hepatitis B virus (HBV). Development of a system supporting efficient replication of clinically resistant HBV strains is imperative, and new antiviral drugs are needed urgently to prevent selection of drug-resistant HBV mutants. A novel fluorinated cytidine analog, NCC (N-cyclopropyl-4′-azido-2′-deoxy-2′-fluoro-β-d-cytidine), was recently shown to strongly inhibit human HBV in vitro and in vivo. This study was designed to evaluate the antiviral activity of NCC against lamivudine-resistant HBV. We generated a stable cell line encoding the major pattern of lamivudine-resistant mutations rtL180M/M204V and designated it “HepG2.RL1”. Immuno-transmission electron microscopic examination and enzyme-linked immunosorbent assay were used to detect secretion of HBV-specific particles and antigens. Quantification of extracellular DNA and intracellular DNA of HepG2.RL1 cells by quantitative real-time polymerase chain reaction revealed >625-fold and >5556-fold increases in the 50% inhibitory concentration of lamivudine, respectively, compared with that for the wild-type virus. The results showed that NCC inhibited DNA replication and HBeAg production in wild-type or lamivudine-resistant HBV in a dose-dependent manner. In conclusion, screening for antiviral compounds active against lamivudine-resistant HBV can be carried out with relative ease using hepG2.RL1 cells. NCC is a potential antiviral agent against wild-type HBV and clinical lamivudine-resistant HBV and deserves evaluation for the treatment of HBV infection.

Comparison of replication competence of wild-type and lamivudine-resistant hepatitis B virus isolates from a chronic hepatitis B patient

In lamivudine-refractory chronic hepatitis B (CHB) patients, discontinuation of lamivudine therapy may lead to loss of lamivudine-resistant hepatitis B virus (HBV) and reappearance of wide-type HBV as dominant strains, yet the underlying mechanism remains unclear. In this study, we cloned wide-type and lamivudine-resistant HBV genomes from the sera of a CHB patient who stopped lamivudine therapy after occurrence of resistant virus and determined the biologic properties of the two isolates in hepatoma cell lines. Sequencing reverse transcriptase region of HBV revealed that the patient developed lamivudine-resistant mutations (rtV173 L, rtL180 M, and rtM204 V) 36 months after the start of lamivudine therapy, and lamivudine-resistant mutants reversed to wild-type after the treatment was stopped for 8 months. Our data showed that the wild-type and mutant isolates had similar transcriptional and translational activity. However, comparison of intracellular and released HBV DNA levels showed that replication efficiency of the mutant virus was approximately 50% of wild-type HBV, while the infectivity of released virus was not affected by the lamivudine-resistant mutations. In conclusion, the reversion of lamivudine-resistant mutants to wild-type HBV after discontinuation of lamivudine in hepatitis B patients may be attributed to better replication fitness of wild-type HBV.

Efficacy of entecavir-based rescue therapy in lamivudine-resistant chronic hepatitis B patients in China: a retrospective study.

Limited studies have investigated the antiviral efficacy of entecavir (ETV)-based rescue therapy in lamivudine (LAM)-resistant chronic hepatitis B (CHB) patients. We retrospectively analyzed the efficacy of entecavir (ETV) monotherapy versus ETV-tenofovir disoproxil fumarate (TDF) combination therapy in 220 LAM-resistant CHB patients. Among 220 patients, 114 patients were treated with ETV monotherapy and 106 were treated with ETV-TDF combination therapy for at least 24 months. There were no significant differences between the two groups in baseline characteristics. During the follow-up of 24 months, virologic response (VR) occurred in 146 (66.4%) patients (58 patients belonged to the ETV monotherapy group and 88 patients belonged to the ETV-TDF combination group). The VR rates were different between the ETV and ETV-TDF groups (32.5% vs. 57.5% at 6 months, 50.0% vs. 77.4% at 12 months; and 50.9% vs. 83.0% at 24 months, P<0.001). In addition, both groups showed no difference in terms of the biochemical and HBeAg response. The rates of viral breakthrough at 6, 12 and 24 months were significantly different between ETV and ETV-TDF groups (2.63%, 4.39% and 9.65% vs. 0.00%, 0.94% and 1.89% at 6, 12 and 24 months, respectively). The ETV-TDF group was superior to the ETV group in achieving a virologic response. Moreover, the ETV-TDF was lower than the ETV group in achieving the initial viral breakthrough and genotypic mutations. Therefore, ETV-TDF combination therapy might be a better regimen than ETV monotherapy in the subgroup of LAM-resistant Chinese patients with CHB.

Role of Cell-Penetrating Peptides in Intracellular Delivery of Peptide Nucleic Acids Targeting Hepadnaviral Replication.

Peptide nucleic acids (PNAs) are potentially attractive antisense agents against hepatitis B virus (HBV), although poor cellular uptake limits their therapeutic application. In the duck HBV (DHBV) model, we evaluated different cell-penetrating peptides (CPPs) for delivery to hepatocytes of a PNA-targeting hepadnaviral encapsidation signal (ε). This anti-ε PNA exhibited sequence-specific inhibition of DHBV RT in a cell-free system. Investigation of the best in vivo route of delivery of PNA conjugated to (D-Arg)8 (P1) showed that intraperitoneal injection to ducklings was ineffective, whereas intravenously (i.v.) injected fluorescein-P1-PNA reached the hepatocytes. Treatment of virus carriers with i.v.-administered P1-PNA resulted in a decrease in viral DNA compared to untreated controls. Surprisingly, a similar inhibition of viral replication was observed in vivo as well as in vitro in primary hepatocyte cultures for a control 2 nt mismatched PNA conjugated to P1. By contrast, the same PNA coupled to (D-Lys)4 (P2) inhibited DHBV replication in a sequence-specific manner. Interestingly, only P1, but not P2, displayed anti-DHBV activity in the absence of PNA cargo. Hence, we provide new evidence that CPP-PNA conjugates inhibit DHBV replication following low-dose administration. Importantly, our results demonstrate the key role of CPPs used as vehicles in antiviral specificity of CPP-PNA conjugates.

Hypophosphatemia predicts a failure to recover from adefovir-related renal injury after dose reduction in lamivudine-resistant hepatitis B patients.

In chronic hepatitis B patients receiving 10mg adefovir, dose reduction is recommended when renal injury appears. However, recovery is not always achieved and markers that recommend switching to another antiviral agent are unknown. We investigated adefovir-related renal injury, recovery after dose reduction, and their predictors. The renal injury in 77 chronic hepatitis B patients receiving 10mg adefovir and recovery after dose reduction to alternate day administration in those with adefovir-related renal injury were assessed. The predictors for >20% estimated glomerular filtration rate (eGFR) decline following treatment with 10mg adefovir and for >20% eGFR recovery after dose reduction were investigated. The adefovir dose was reduced in 26 patients (34%) at 59 ± 30 (mean ± standard deviation) months of 10mg adefovir treatment because of decreases in eGFR (cumulative incidence 27%), serum phosphorus (9%), and uric acid (16%) levels, and increases in alkaline phosphatase (20%), bone type alkaline phosphatase (18%), urinary α1-microglobulin (18%), and urinary N-acetyl-β-D-glucosaminidase (18%) levels. The only significant predictor for >20% eGFR decline was age ≥50years at the start of 10mg adefovir treatment. The cumulative eGFR recovery rate was 42% at 42 ± 27months after dose reduction, and ≥2.5mg/dL serum phosphorus level at dose reduction was the only significant predictor for >20% eGFR recovery after dose reduction. Patients aged ≥50years are predisposed to adefovir-related renal injury and switching to another antiviral agent rather than adefovir dose reduction is recommended when hypophosphatemia is observed.

Emtricitabine for adults with lamivudine-resistant chronic hepatitis B virus infection

Emtricitabine for adults with lamivudine-resistant chronic hepatitis B virus infection

Probable lamivudine-resistant hepatitis B infection in children: A case report involving two Nigerian siblings at Ahmadu Bello University Teaching Hospital, Shika, Zaria

Currently, up to 65 million Africans have chronic hepatitis B (CHB) infection, with the majority acquiring it in the first 5 years of life and living in Nigeria. Lamivudine is generally accepted for use as the first-line treatment of CHB in children under the age of 12 years. However, the drug has a low genetic barrier to resistance. Lamivudine-resistant CHB is a serious challenge in resource-limited settings. This case report is intended to study a probable lamivudine-resistant CHB in two siblings aged 9 and 11 years and to highlight the challenges of managing CHB in children from resource-limited settings. There is a need to develop national guidelines for the management of CHB in children. Other suitable antiviral agents should be subsidized, so that they could be used to salvage lamivudine-resistant cases.

Tenofovir disoproxil fumarate (TDF) vs. emtricitabine (FTC)/TDF in lamivudine resistant hepatitis B: A 5-year randomised study

Long-term treatment with tenofovir disoproxil fumarate (TDF) alone, or in combination with emtricitabine (FTC) is associated with sustained viral suppression in patients with lamivudine resistant (LAM-R) chronic hepatitis B (CHB). LAM-R CHB patients were randomised 1:1 to receive TDF 300mg or FTC 200mg and TDF 300mg once daily in a prospective, double blind, study. The proportion of patients with plasma hepatitis B virus (HBV) DNA<69IU/ml (<400copies/ml) at week 96 (primary efficacy endpoint) was reported previously. Here we present week 240 follow-up data. Overall, 280 patients were randomised to receive TDF (n=141) or FTC/TDF (n=139), and 85.4% completed 240weeks of treatment. At week 240, 83.0% of patients in the TDF arm, and 82.7% of patients in the FTC/TDF treatment arm had HBV DNA<69IU/ml (p=0.96). Rates of normal alanine aminotransferase (ALT) and normalised ALT were similar between groups (p=0.41 and p=0.97 respectively). Hepatitis B e antigen loss and seroconversion at week 240 were similar between groups, (p=0.41 and p=0.67 respectively). Overall, six patients achieved hepatitis B surface antigen (HBsAg) loss and one patient (FTC/TDF arm) had HBsAg seroconversion by week 240. No TDF resistance was observed up to week 240. Treatment was generally well tolerated, and renal events were mild and infrequent (∼8.6%). The mean change in bone mineral density at week 240 was -0.98% and -2.54% at the spine and hip, respectively. TDF monotherapy was effective and well tolerated in LAM-R CHB patients for up to 240weeks. The goal of oral antiviral treatment for chronic hepatitis B (CHB) is to achieve and maintain undetectable HBV DNA levels. Treatment options with enhanced potency, and low risk of resistance development for patients infected with lamivudine resistant (LAM-R) HBV are required. Tenofovir disoproxil fumarate (TDF) monotherapy was effective and well tolerated without TDF resistance development in CHB patients with LAM-R, for up to 240weeks. Clinical trial number: NCT00737568.

Continued lamivudine for adults with lamivudine-resistant chronic hepatitis B virus infection

Continued lamivudine for adults with lamivudine-resistant chronic hepatitis B virus infection

Interferon for people with lamivudine-resistant chronic hepatitis B virus infection

Interferon for people with lamivudine-resistant chronic hepatitis B virus infection

Development and validation of an oligonucleotide ligation assay to detect lamivudine resistance in hepatitis B virus

Treatment of chronic hepatitis B virus (HBV) infection with lamivudine-monotherapy rapidly selects mutant variants in a high proportion of individuals. Monitoring lamivudine resistance by consensus sequencing is costly and insensitive for detection of minority variants. An oligonucleotide ligation assay (OLA) for HBV lamivudine-resistance was developed and compared to consensus sequencing. Both assays detected drug resistance mutations in 35/64 (54.7%) specimens evaluated, and OLA detected minority mutants in an additional six (9.4%). OLA may offer a sensitive and inexpensive alternative to consensus sequencing for detection of HBV drug resistance in resource-limited settings.

Clinical investigation of entecavir combined with adefovir dipivoxil in the treatment of lamivudine-resistant hepatitis B cirrhosis

Objective To investigate the clinical effect of entecavir combined with adefovir dipivoxil in the treatment of lamivudine-resistant hepatitis B cirrhosis, to provide a reference for clinical treatment. Methods 117 cases of liver cirrhosis with lamivudine resistance were selected, they were divided into the control group and the observation group according to treatment.60 cases in the observation group used entecavir and adefovir combination therapy, 57 cases in the control group was given lamivudine combined with adefovir dipivoxil.The HBeAg conversion rate, HBV-DNA negative rate, liver function, liver function Child-pugh score were compared between two groups. Results After treatment for 24 weeks, 48 weeks, the HBV-DNA negative conversion rates in the observation group were 75.00%, 95.00%, which were higher than those in the control group, the differences were statistically significant (χ2=4.251, P=0.024; χ2=4.535, P=0.018). In the observation group, ALB, ALT, TBiL, PT improved better than the control group, the differences were statistically significant(t=4.229, P=0.025; t=6.214, P=0.008; t=5.514, P=0.014; t=5.233, P=0.017). After treatment, CTP of the observation group was (7.15 ± 1.05) points, which was significantly lower than the control group (8.86 ± 1.47) points, the difference was statistically significant (t=5.874, P=0.010). The incidence rate of adverse reactions between the two groups showed no statistically significant difference(P>0.05). Conclusion Entecavir combined with adefovir dipivoxil therapy has good effect for lamivudine-resistant liver cirrhosis, which will help to improve liver function, inhibit HBV replication, it is worthy of clinical application. Key words: Liver cirrhosis; Entecavir; Adefovir dipivoxil; Lamivudine

Efficacy of tenofovir-based rescue therapy in patients with lamivudine-resistant hepatitis B virus: A systematic review and meta-analysis

Currently, there are no conclusive results on the efficacy of Tenofovir disoproxil fumarate (TDF) monotherapy in chronic hepatitis B (CHB) patients with lamivudine-resistant (LAM-R). The aim of this study was to compare the efficacy between TDF and TDF-based combination therapy against LAM-R HBV in CHB patients. Randomized and non-randomized control trials directly comparing TDF and TDF-based therapy for treatment of LAM-R CHB patients, were searched in Pubmed, Medline, EMBASE, database up to June 15, 2015. The data were analyzed with Review Manager (v.5.3). Five articles (683 patients in total) met entry criteria. The overall efficacy of tenofovir-based combination therapy was not significantly better with regard to the rates of virological response (85.5% vs. 81.5%; RR=0.95, 95%CI=0.88-1.03, P=0.25), ALT normalization (61.9% vs.72.0%; RR=1.18, 95%CI=0.96-1.44, P=0.11) and HBeAg loss (17.0% vs. 18.1%; RR=1.40, 95%CI=0.78-2.49, P=0.26) compared with TDF monotherapy through 48-week treatment. Additionally, subgroup analysis showed that no significant difference was determined as TDF group compared to TDF-based group at 48weeks, in terms of rates of HBV DNA undetectability, ALT normalization and HBeAg loss in the treatment of LAM-R patients with prior failure of LAM monotherapy. Moreover, the rates of HBV DNA suppression between groups were similar through 24 or 48weeks of treatment in LAM-R patients with prior failure of LAM/ADV therapy. TDF monotherapy is as effective as TDF-based combination therapy in maintaining viral suppression in LAM-R patients with prior failure of LAM or LAM/ADV therapy.

Emergence of lamivudine-resistant hepatitis B virus during combination antiretroviral therapy that includes lamivudine for patients co-infected with HIV and hepatitis B virus in China: a 2-year pilot cohort study

BackgroundIn China, more than 10% of individuals infected with HIV are carriers of hepatitis B virus (HBV). Patients infected with HIV-1 are usually given combination antiretroviral therapy that includes lamivudine (ART-3TC) as a reverse-transcriptase inhibitor. Previous studies from some developed counties showed that, in ART-3TC, 3TC-resistant HBV progressively emerges at a rate of 15–20% per year in patients co-infected with HIV-1 and HBV. We investigated the occurrence of 3TC-resistant HBV during ART-3TC for double infections and tested the use of tenofovir disoproxil fumarate (TDF) as an additional reverse-transcriptase inhibitor (ART-3TC/TDF) in a 2-year cohort study in China. Methods172 plasma samples from 43 Chinese patients co-infected with HIV-1 and HBV (from 134 cases positive for HBsAg) were examined for the prevalence of 3TC-resistant HBV by direct sequencing of PCR products covering the HBV reverse transcriptase gene. The patients were randomised (7:1), using a randomised number table, to receive either ART-3TC or ART-3TC/TDF (because ART-3TC/TDF was not a standard first-line treatment in China at that time). HIV-1 RNA and HBV DNA loads were detected at baseline, 12 weeks, 48 weeks, and 96 weeks from plasma samples. All samples were tested for HIV and HBV drug resistance against 3TC. The efficacy of viral suppression and incidence of drug-resistance mutation were compared between the two groups. This study was approved by the ethics committee at Peking Union Medical College Hospital, Beijing, China, and registered at ClinicalTrials.gov (identifiers NCT00618176 and NCT00872417). Patients were enrolled in Chinese cohort studies (National Key Technologies R&D Program for the tenth and 11th Five-year Plans). FindingsMedian HIV-1 RNA loads of the ART-3TC group and the ART-3TC/TDF group significantly decreased from 26 205 copies/mL (IQR 917; p<0·0001) to fewer than 20 copies/mL (<20; p=0·0278) and from 58 972 copies/mL (3395; p<0·0001) to fewer than 20 copies/mL (<20; p=0·0207), respectively, after 96 weeks of treatment. Median HBV DNA loads also significantly decreased from 3·4 × 106 IU/mL (IQR <20; p<0·0001) to less than 20 IU/mL (<20; p<0·0001) in the ART-3TC group and from 2·7×105 IU/mL (399; p<0·;0001) to less than 20 IU/mL (<20; p=0·0152) in the ART-3TC/TDF group after 96 weeks of treatment. Although median HIV-1 viral load did not differ significantly (p>0·05) between the two groups after 96 weeks, the rate of HIV-1 RNA decline in the ART-3TC/TDF group was slightly lower than that in the ART-3TC group (RNA load range <20–905 copies/mL vs <20–4932 copies/mL at 12 weeks, <20–157 copies/mL vs <20–398 copies/mL at 48 weeks). ART-3TC/TDF had a higher efficacy than ART-3TC in controlling amounts of HBV. At 96 weeks, the median HBV DNA load was less than 20 IU/mL (IQR <20–1·2 × 108) in the ART-3TC group, suggesting that some patients had rebound or uncontrolled HBV DNA loads, but no such case was reported in the ART-3TC/TDF group, the median HBV DNA load of which was <20 IU/mL (<20; p=0<0119). Ten (23%) of 43 patients, all from the ART-3TC group, had HBV breakthrough after 96 weeks of treatment, and eight of these patients were resistant to ART-3TC (60% tr204 mutation and 80% rt180 mutation; appendix). InterpretationAlthough both regimens were able to control replication of HIV and HBV during the 2-year treatment, our findings showed that, in China, ART-3TC is likely to lead to the emergence of 3TC-resistant HBV, lending support to the benefit of the inclusion of TDF for patients co-infected with HIV-1 and HBV.

Adefovir dipivoxil-induced development of osteomalacia and Fanconi syndrome during the treatment of hepatitis B virus (HBV)-related cirrhosis

Adefovir dipivoxil is a nucleotide analog reverse transcriptase inhibitor used to treat adult patients affected by HBeAg-positive and HBeAg-negative chronic hepatitis B and with clinical evidence of lamivudine-resistant hepatitis B virus (HBV). Adefovir administered at a dosage of 10 mg/day is generally well tolerated, even if renal toxicity, type Fanconi syndrome, was reported during long-term treatments.We report a case of osteomalacia with Fanconi syndrome and pathologic fracture of the femur related to long-time (67 months) adefovir treatment (10 mg/day) in a patient with compensated hepatitis B virus (HBV) cirrhosis (Child 5A) and with a previous normal renal function (estimated Glomerular Filtration Rate before adefovir = 78.26 ml/min/1.73 m2; during adefovir treatment = 57.38 ml/min/1.73 m2). The patient was switched to entecavir at a dose of 1 mg/day, with both suppression of viremia and improvement of osteomalacia and Fanconi syndrome; the patient’s follow-up is still ongoing after 22 months.

Hepatitis B virus genotyping among chronic hepatitis B individuals with resistance to Lamivudine in shahrekord, iran.

Background:Hepatitis B infection, caused by hepatitis B Virus (HBV), is one of the major global public health problems. Hepatitis B Virus genotypes appear to show varying geographic distribution with possible pathogenic and therapeutic differences. Knowledge of HBV genotypes is very important for clinical treatment. Lamivudine is a nucleoside analogue that is clinically used to treat chronic hepatitis B infection. However, the main problem with the application of lamivudine is the development of viral resistance to the treatment with this anti viral drug. Besides, it has been suggested that lamivudine -resistant HBV may be genotype dependent. However, HBV genotype distribution and the biological relevance in this region are poorly understood.Objectives:The current study aimed to determine hepatitis B genotypes and their correlation with lamivudine- resistant HBV frequency among patients with chronic hepatitis B from Shahrekord, Iran.Methods and Materials:Hepatitis B virus DNA was detected by conventional PCR in some of the serum samples obtained from HBsAg-positive Chronic Hepatitis B (CHB) patients who were referred to Health Centers of Shahrekord for routine monitoring of the disease. Subsequently, using real-time PCR, the DNA samples were used for genotyping and analysis of resistance to lamivudine.Results:The DNA was detected in 23 out of 116 (19.82%) of the studied samples. Genotypes D and C were found in 17 out of 23 (73.9%), and in 6 out of 23 (26.1%) of the samples, respectively. To the authors’ best knowledge, the current study is the first report on isolation of Genotype C from Iran. Two out of 17 (11.76%), and 6 out of 6 (100%) of genotypes D and C were resistant to lamivudine, respectively. Resistance to this drug was significantly different between genotypes C and D (P <0.001).Conclusions:In addition to genotype D, other lamivudine resistant hepatitis B genotypes might be distributed in Iran.

Generation of a human hepatoma cell line supporting efficient replication of a lamivudine resistant hepatitis B virus

Emergence of lamivudine (LAM) resistance causes treatment failure in patients with chronic hepatitis B and compromise the efficacy of subsequent salvage therapies with other nucleot(s)ide analogs (NAs). Establishment of cell-based assays supporting LAM-resistant hepatitis B virus (HBV) replication will not only provide tools for investigating the replication property, but also screening for antiviral agents efficiently inhibiting the replication of LAM-resistant HBV variants. Accordingly, a human hepatoma (HepG2)-derived cell line was established by stable transfection of a plasmid containing a 1.2 unit length of HBV genome harboring rtL180M and rtM204V mutations that confer LAM resistance. In addition to support efficient viral genome replication, the cell line also produces high levels of HBV virions and subviral particles. As expected, HBV DNA replication in this cell line is completely resistant to lamivudine, but sensitive to adefovir (ADV), entecavir (ETV) and tenofovir (TDF). The cell line is suitable for screening for antiviral agents that inhibit LAM-resistant HBV replication and inhibitors of HBsAg biosynthesis and secretion, which may reduce HBsAg antigenemia and ultimately help to restore host antiviral immune response against HBV and cure chronic HBV infection.