Negative Hepatitis B Virus DNA Research Articles

An interesting case of isolated false-negative hepatitis B surface antigen in a blood donor.

An important requirement for a hepatitis B surface antigen (HBsAg) screening assay is reliable detection of HBsAg mutant forms, especially in blood donation. Here we investigate and describe the case of an isolated false-negative result of commercial serology HBsAg screening assay of a blood donor. The current donation was routinely tested for HBsAg and hepatitis B virus (HBV) DNA in the mini-pool mode nucleic acid testing (MP-NAT of six samples), and further evaluated by individual donation ID-NAT. Finally, it was quantified and sequenced. All previous donations were found to have negative HBsAg and HBV DNA, as also the subsequent sample taken 3 months after the marked donation. The current donation of the 53-year-old unvaccinated female with 14 previous donations was initially HBsAg negative and HBV DNA (MP-NAT) positive. Further testing showed HBsAg positive using other HBV serological assays, antibodies to HBV core antigen immunoglobulin Mpositive and HBV DNA ID-NAT positive, and contained 200 IU/mL of HBV DNA. The implicated donation carried genotype D strains, subtype ayw2 (F83S, V96A, V190A, L193S, I195T, L213S, F220L). The mutations in three positions, namely amino acids T118A, P120T, and P127T, were proven subsequently. This unique mutation combination near the target epitope of one of the immunoassay monoclonals is a possible cause of the reduced analytical sensitivity of the serology assay.

Effect of previous infection with hepatitis B virus on the incidence of hepatocellular carcinoma after sustained virologic response in patients with chronic hepatitis C virus infection.

Previous infection with hepatitis B virus (HBV), which is assessed by HBV core antibody (HBcAb) or surface antibody (HBsAb) titres, has reportedly been associated with an increased risk of developing hepatocellular carcinoma (HCC). We investigated the influence of previous HBV infection on the incidence of HCC in patients with hepatitis C virus (HCV) infection who achieved eradication of HCV, that is sustained virologic response (SVR). Both HBcAb and HBsAb were measured in a total of 1214 patients with HCV infection who had not been coinfected with HBV, as determined by both negative HBs antigen and HBV DNA, and in whom SVR was confirmed. Patients were followed up for a median of 5.7 years, and the incidence of post-SVR HCC was compared based on HBcAb and/or HBsAb. In both univariate and multivariate analyses, the incidence of post-SVR HCC did not differ based on the presence of HBcAb or HBsAb. In conclusion, previous HBV infection has no impact on the incidence of HCC in patients with HCV after SVR.

Hepatocellular Carcinoma and Hepatitis: Advanced Diagnosis and Management with a Focus on the Prevention of Hepatitis B-Related Hepatocellular Carcinoma.

Though the world-wide hepatitis B virus (HBV) vaccination program has been well completed for almost thirty years in many nations, almost HBV-related hepatocellular carcinoma (HCC) occurs in unvaccinated middle-aged and elderly adults. Apparently, treating 80% of qualified subjects could decrease HBV-related mortality by 65% in a short period. Nevertheless, globally, only 2.2% of CHB patients undergo antiviral therapy. The HBV markers related to HCC occurrence and prevention are as follows: the HCC risk is the highest at a baseline of HBV DNA of 6-7 log copies/mL, and it is the lowest at a baseline of an HBV DNA level of >8 log copies/mL and ≤4 log copies/mL (parabolic, and not linear pattern). The titer of an HBV core-related antigen (HBcrAg) reflecting the amount of HBV covalently closed circular DNA (ccc DNA) in the liver is related to HCC occurrence. The seroclearance of HBs antigen (HBsAg) is more crucial than HBV DNA negativity for the prevention of HCC. In terms of the secondary prevention of hepatitis B-related HCC involving antiviral therapies with nucleos(t)ide analogues (NAs), unsolved issues include the definition of the immune-tolerant phase; the optimal time for starting antiviral therapies with NAs; the limits of increased aminotransferase (ALT) levels as criteria for therapy in CHB patients; the normalization of ALT levels with NAs and the relation to the risk of HCC; and the relation between serum HBV levels and the risk of HCC. Moreover, the first-line therapy with NAs including entecavir (ETV), tenofovir disoproxil fumarate (TDF), and tenofovir alafenamide (TAF) remains to be clarified. Discussed here, therefore, are the recent findings of HBV markers related to HCC occurrence and prevention, unsolved issues, and the current secondary antiviral therapy for the prevention of HBV-related HCC.

Atypical serologic profiles of hepatitis B virus infection across clinical cohorts of patients in Southwestern Nigeria

ABSTRACT Hepatitis B virus (HBV) infection follows a natural course of events predicted by a dynamic interaction between viral antigen and the host immune system, which forms the basis for HBV serological diagnosis. These interactions may deviate from the typical serologic patterns. This study investigates the types of atypical HBV serologic profiles (AHBSP) across clinical cohorts of patients with HBV infection in southwestern Nigeria. This is a cross-sectional, hospital-based, multi-centered study. Patients’ sera were analyzed for HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc IgM, and anti-HBc IgG by ELISA from 279 study participants attending selected gastroenterology clinics between August 2019 and December 2020. The prevalence of atypical HBV serologic profiles was 27% (n = 76). The mean age of patients was 35.7 ± 11.2 years. The gender distribution involved 183 females (65.6%) and 96 males (34.4%). Across clinical cohorts of patients with atypical serologic profiles, HBeAg Negative, anti-HBe positive with detectable HBV DNA had the highest prevalence of 21% followed by isolated anti-HBc antibody positive, HBsAg negative and detectable HBV DNA, 5%. The atypical serologic profiles, HBeAg positive, HBsAg negative with detectable HBV DNA and concurrent anti-HBs with HBsAg, had the lowest prevalence, 0.4%, respectively. This study identified the considerable presence of atypical HBV serologic profiles across clinical cohorts of HBV infection in southwestern Nigeria.

Comparison of S gene mutations in patients with occult and chronic hepatitis B virus infection

Background and aimThis study aimed to detect mutations in the HBV S gene and evaluate their relationship to occult hepatitis B virus (HBV) infection (OBI). MethodsThe study included 32 patients with negative serum HBsAg and HBV DNA who underwent liver biopsy due to different clinical indications defined as the OBI group and 32 patients who underwent liver biopsy due to chronic hepatitis B (CHB) as the comparison group. The HBV S gene region was amplified by Nested PCR, and Sanger sequencing was performed. ResultsAt least one amino acid (aa) mutation was detected in the major hydrophilic region (MHR) of the HBV S gene in 14/32 (43.75%) of the patients with OBI and 8/32 (25.0%) with CHB. The genotype of all patients with OBI and CHB was HBV/D. Although 9 (28.1%) of the cases with OBI had sub-genotype HBV/D3, none of the patients with CHB had sub-genotype HBV/D3. Unlike patients with CHB, L15*, D33N, Q51P, V63F, L91I, P108S, T115I, P120L, T125M, Q129H, T189I, L216F, P217L mutations were detected in the HBV S gene in OBI cases. Also, P127T aa polymorphism was frequently detected. Mutation frequency in the HBV S gene in the major hydrophilic region (MHR) was higher in patients with OBI with sub-genotypes HBV/D3 and D2 than those with HBV/D1 and those with serotype HBV/ayw3 compared to those with HBV/ayw2 (p < 0.05). ConclusionsSub-genotypic-specific mutation patterns were seen in the “a” determinant region and T helper cell epitopes of HBsAg, especially in the C-terminus domain; this may be associated with OBI.

Role of hepatosteatosis in HBsAg seroconversion in HBeAg-negative chronic hepatitis B patients.

In chronic Hepatitis B virus (HBV) infection, certain individual and viral characteristics such as advanced age, presence of hepatic steatosis (HS), normal ALT levels, initially negative HBeAg and HBV DNA, and genotype of the virus are associated with HBsAg seroclearance and seroconversion. Herein, we report the results of our study evaluating the association between hepatosteatosis and HbsAg seroconversion. The clinical and biochemical data of patients with CHB and hepatosteatosis (HS) (HBsAg seroconversion, n:52, and non-HbsAg seroconversion, n:352), and the rate of development of HBsAg seroconversion were evaluated. We collected data from 404 patients with HBeAg negative CBH (mean age ± SD: 36.2±11years; 223 [55.2%] men, 181 [44.8%] women). The mean age at diagnosis of disease was 36.2±11years. The mean duration of the disease was 10.6±7years. Seroconversion developed in 52 patients (12.8%) with serum HBsAg positive (mean ± SD: 12.7±5.8). Elderly age and the duration of disease time were significantly associated with seroconversion (P<.001). The presence of serum HBsAg seroconversion was significantly associated with hepatosteatosis (OR: 3.06, 95% CI 1.64-5.71, P<.01). Serum HBsAg seroconversion was more frequent in patients with mild HS than patients with moderate-severe HS (P=.04). In multivariate regression analysis, the presence of HS was found to be an independent factor predicting the development of HBsAg seroconversion (OR: 2.07 95% GA:1.07-4.0 P=.03). The presence of mild HS in HBeAg negative chronic hepatitis B patients contributes to HBsAg seroconversion. Further studies are required to better understand the relationship between steatosis and HBsAg seroconversion.

Auxiliary Liver Graft Can Be Protected From HBV Infection in HBsAg Positive Blood Circulation.

Auxiliary grafts have a high risk of Hepatitis B virus (HBV) infection in patients with chronic HBV-related diseases. Hepatitis B virus-related auxiliary partial orthotopic liver transplantation (APOLT) cases were reviewed to show the results of current methods to block native-to-graft HBV transmission. Three patients received APOLT for HBV-related liver cirrhosis and a recurrent upper gastrointestinal hemorrhage between April 2015 and January 2017 by the liver transplant team of Beijing Friendship Hospital affiliated with Capital Medical University. All three patients were positive for HBV surface antigen (HBsAg) and had a negative HBV DNA test result before transplantation. After auxiliary transplantations, HBsAg was found to be positive in two patients and negative in one patient. To avoid graft infection of HBV, entecavir-based therapy was employed and the remnant native livers of the recipients were removed 51–878 days after liver transplantation. Then, serum conversions of HBsAg were found in all three cases. For the first time, this case series shows the possibility of blocking the transmission of HBV from a native liver to a graft in auxiliary transplantation by entecavir-based therapy. Among the cases, a left lobe graft was successfully implanted as a replacement of the right lobe of the recipient, which is also discussed.

Clinical Features and Resistance to Entecavir Monotherapy of Patients with Hepatitis B.

Aim Hepatitis B virus (HBV) infection is a major public health concern worldwide. Entecavir (ETV), a first-line nucleos(t)ide analogue (NA) for HBV, has a low risk of resistance. We evaluated the efficacy of ETV monotherapy, ratio of ETV-resistant, and the clinical features of patients with ETV resistance. Methods A total of 130 patients (72 males, 58 females; mean age, 61 ± 15 years) were divided into a NA-naïve group (n = 108) and NA-experienced group (n = 22). We examined the clinical outcomes of ETV monotherapy and associated factors. We also assessed the clinical features of 15 patients with resistance to ETV (mean, 51.0 ± 27.4 weeks). Results Among the 130 patients, 94.1% achieved ALT normalization and 63.6% achieved serum HBV DNA negativity after ETV monotherapy for 96 weeks. Of the patients in the NA-naïve group, 93.1% and 60.4% achieved ALT normalization and HBV DNA negativity, respectively. Of the patients in the NA-experienced group, 100% and 74.9% achieved ALT normalization and HBV DNA negativity, respectively. Compared to patients on ETV continuously, 15 ETV-resistant patients had a higher baseline HBV viral load. There was a significant difference in the time to HBV DNA negativity, but not ALT normalization after ETV monotherapy in these groups. Rescue treatment with other NAs led to ALT normalization in all of these patients, but not HBV DNA negativity. Conclusions ETV monotherapy has a long-term clinical efficacy. While some patients especially with HBV DNA high viral load developed ETV resistance, rescue treatment led to ALT normalization in these patients.

Hepatitis B Virus Seropositivity in Chronic Myeloid Leukemia Patients Receiving Tyrosine Kinase Inhibitors: Preliminary Results of a Single-Center Cross-Sectional Study

Background: Hepatitis B virus (HBV) infection is a global public health issue, and Turkey is intermediately endemic with 4.5% and 30.6% HbsAg and anti-HBc IgG prevalences, respectively. The HBV infection can be problematique in patients (pts) with hematological malignancies. Tyrosine kinase inhibitors (TKIs) are the mainstay of chronic myeloid leukemia (CML) management, and screening of CML pts for HBV prior to TKI initiation and the management of isolated anti-HBc IgG positivity among these pts are controversial.Aim: To evaluate the frequency and outcome of HbsAg and anti-HBc IgG positivity in CML pts receiving TKIs.Patients and methods: Seventy-four pts with CML in chronic phase (CML-CP) were tested for HBV markers (HbsAg, anti-Hbs, anti-Hbc IgG, HBV DNA) and serum alanine aminotransferase (ALT) levels out of a total of 240 pts as a part of an ongoing study. Pts' characteristics, TKI therapies and durations, response rates, and follow-up periods were noted retrospectively.Results: Pts' characteristics are summarized in Table 1. Fifty-six pts received imatinib (IM) and nilotinib (NIL) was administered in one patient as first-line treatments, whereas the remaining 17 pts had two or more lines of TKI. Median duration of follow-up was 104 months (mos) (range, 3-215 mos). Only one patient was screened for HBV prior to TKI therapy. The median time from TKI initiation to HBV screening was 98 mos (range, 2-179 mos).Fourty-three pts were either HBV naive or anti-Hbs negative. Nineteen pts had anti-Hbc IgG positivity, of whom 10 were also positive for anti-Hbs. Among anti-Hbc IgG positive cases, HBV DNA was negative in twelve pts, and the HBV DNA results of the remaining 7 cases are still pending. None of the pts with isolated anti-Hbc IgG positivity received antiviral prophylaxis for HBV. Four pts (5%) had elevated ALT levels at the time of analysis. One of these pts was HbsAg positive (ALT: 65 IU/ml) (Table 2, case 1); the other was anti-Hbc IgG positive but had negative anti-Hbs and HBV DNA (ALT: 54 IU/ml) receiving IM, whereas the remaining two were HBV negative. Of these two cases, one had grade I (ALT: 57 IU/ml) and the other had grade II (ALT: 101 IU/ml) hepatotoxicity most probably due to dasatinib and NIL, respectively.Two pts were HbsAg positive, and the clinical features of these cases are shown in Table 2. First case had elevated ALT levels (174 IU/ml), positive HbeAg and high HBV DNA which were detected one mo after IM initiation. Tenofovir was started and he had decline in HBV DNA. The patient is still under IM with MMR. The other case experienced HBV reactivation 28 months after IM therapy. With lamivudine treatment, HBV DNA became negative and he is still under IM maintaining MMR.Discussion: The risk of HBV reactivation under TKI therapy and thus, the necessity of screening for HBV prior to TKI initiation are still debating. HBV screening before TKI therapy is not recommended in the ESMO and ELN guidelines, whereas the most recent NCCN guideline and EMA document recommended testing for HBV infection before TKI administration due to some case reports of HBV reactivation after TKI initiation. In our patient cohort the percentages of HbsAg and anti-Hbc IgG positivities were 3% and 26% among the tested cases which were consistent with the prevalences for our general population. Although the nature of our study is cross-sectional and the results are preliminary, the risk of HBV reactivation with TKI use seems to be low in pts with resolved HBV infection and with isolated anti-Hbc IgG positivity and negative HBV DNA among our cohort. Longitudinal studies with serial testing for HBV markers are needed in order to answer these questions in CML pts receiving TKI therapy. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Comparison of HBV-DNA Levels with Biochemical and Microbiological Parameters for Chronic Hepatitis Evaluation, Bursa, Turkey

IntroductionHBV-DNA levels are used to diagnose chronic hepatitis B (CHB) disease, determine the infection phase, decide on the treatment, and determine the disease course. We aimed to compare the microbiological and biochemical parameters of patients followed up with chronic hepatitis B pre-diagnosis in our hospital according to their HBV-DNA levels.MethodsHBV-DNA levels were analyzed with the Real-Time PCR method in blood samples of 500 pre-diagnosed CHB patients between February-June 2018, retrospectively. The biochemical parameters of the patients were measured by an automatic biochemical immunoassay analyzer (Beckman Coulter DXI 800, USA), and the microbiological parameters of patients were determined by an automatic analyzer (Roche Cobas 6000, Germany). The differences between the values of biochemical and microbiological parameters of patients were determined according to HBV-DNA.ResultsMean corpuscular volume (MCV) was higher in patients with hepatitis B virus-DNA (HBV-DNA)>20000 IU/mL than patients with negative HBV-DNA, HBV-DNA 20000 IU/mL (P 20000 IU/mL than patients with HBV-DNA 2000-20000 IU/mL and HBV-DNA 20000 IU/mL (P<0.05, P<0.05, P<0.05). Albumin was individually correlated with the HBV-DNA by 2.9%, negatively (P<0.01).ConclusionMCV, GGT, albumin, HBsAg levels might be used as indicators to diagnose CHB disease, establish the infection phase, decide the treatment, and determine the course of the disease together with HBV-DNA levels.

Diagnostic value of the \u03b3-glutamyltransferase and alanine transaminase ratio, alpha-fetoprotein, and protein induced by vitamin K absence or antagonist II in hepatitis B virus-related hepatocellular carcinoma

Hepatocellular carcinoma is a common type of malignancy with a poor prognosis. Identification and utilisation of markers for monitoring and diagnosis are urgently needed. Alpha-fetoprotein (AFP) and Protein Induced by Vitamin K Absence or Antagonist-II (PIVKA-II) have been proved to be efficient biomarkers for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). The combination of the two markers could improve the detection rate. However, these indicators cannot meet the need of clinical diagnosis.It is necessary to discover novel serological markers and more cost-effective, appropriate combination of these markers for the diagnosis and surveillance of HBV-related HCC. Accordingly, in this study, we aimed to evaluate the diagnostic value of γ-glutamyltransferase (γ-GT) to alanine amino transferase (ALT) ratio alone or in combination with AFP and PIVKA-II for HBV-related HCC. 234 patients with HBV-related HCC and 396 patients with chronic hepatitis B (CHB) were enrolled in this study and approved by the institutional review board. Our results showed levels of AFP and PIVKA-II, and γ-GT/ALT ratio in cases with early-stage HCC, HCC, HCC plus HBV DNA positivity, and HCC plus HBV DNA negativity were higher than those in the corresponding CHB control group. Additionally, the levels of serum AFP and PIVKA-II, and the γ-GT/ALT ratio were positively correlated with tumour sizes in patients with HBV-related HCC. The areas under the ROC curves (AUROCs) of the γ-GT/ALT ratio in patients with early-stage HCC, HCC, HCC plus HBV DNA positivity, and HCC plus HBV DNA negativity were 0.795, 0.846,0.855, and 0.837, respectively; AUROCs of combination of the γ-GT/ALT ratio and PIVKA-II were 0.858, 0.928, 0.948, and 0.902, respectively; AUROCs of combination of the γ-GT/ALT ratio and AFP were 0.822, 0.886, 0.896, and 0.873, respectively;AUROCs of combination of the γ-GT/ALT ratio and PIVKA-II with AFP were 0.857, 0.928, 0.946, and 0.907, respectively, and AUROCs of combination of PIVKA-II and AFP were 0.804, 0.904, 0.942, and 0.863, respectively. In conclusion, the γ-GT/ALT ratio was a useful biomarker for the diagnosis of HBV-related HCC and that the combination of AFP and PIVKA-II with the γ-GT/ALT ratio could improve the diagnostic value of these biomarkers for HBV-related HCC. Moreover, the ratio of γ-GT/ALT may be a useful index in monitoring patients for progression of HBV-related HCC.

Возможность использования количественного определения HBsAg при хронической HBV-инфекции с дельта-агентом

We have conducted a pilot study to identify the association between the HBsAg level and activity of the pathological process in the liver, as well as possibility of quantitative assessment of HBsAg for monitoring chronic liver diseases caused by hepatitis B virus (HBV) and hepatitis D virus (HDV). Objective. To assess the possibility of using HBsAg levels as a predictor of disease activity and prognosis in patients with chronic HBV infection with delta-agent. Patients and methods. We analyzed serum specimens from 30 patients with HDV and HBV co-infection. Among 15 patients with chronic hepatitis B with delta-agent, there were 5 HBV DNA positive and 10 HBV DNA negative. Among patients with liver cirrhosis, HBV DNA was detected in 11 individuals, while 4 individuals had undetectable HBV DNA levels. Results. We found that mean HBsAg level in patients with chronic HBV infection and negative HBV DNA was 1.9 ± 0.56 IU/mL. Mean HBsAg level in patients with chronic HBV infection with delta-agent and positive HBV DNA was 4.3 ± 0.62 IU/mL (p &lt; 0.05). High HBsAg levels correlated with elevated ALT in patients with chronic hepatitis B and delta-agent. Patients with liver cirrhosis caused by HDV had normal ALT levels, but elevated bilirubin concentrations regardless of HBV DNA presence and HBsAg level. Conclusion. High levels of HBsAg can be considered as a predictor of active disease in patients with chronic HBV infection with delta-agent and also a marker of transformation of chronic hepatitis B with delta-agent into liver cirrhosis. Key words: chronic hepatitis B with delta agent, liver cirrhosis, enzyme-linked immunosorbent assay, HBsAg levels, polymerase chain reaction

Low prevalence of occult hepatitis B virus infection in chronic haemodialysis and kidney transplant patients

Occult hepatitis B virus infection (OBI) is defined as the presence of hepatitis B virus (HBV) DNA in serum and/or liver in HBsAg-negative patients. We investigated the prevalence of OBI in large chronic haemodialysis (CHD) and kidney transplant recipients (KTxR) cohorts, including determination of HBV DNA in peripheral blood mononuclear cells (PBMCs). HBV DNA was determined in both serum and PBMCs in 417 CHD patients, 417 KTxR, 20 HBsAg-positive non-CHD non-KTx patients (positive controls) and 40 HBsAg-negative healthy subjects (negative controls). Chronic haemodialysis group: two of 376 patients were HBsAg-positive. The 374 HBsAg-negative patients tested negative for HBV DNA in both serum and PBMCs. KTxR group: 14 of 417 patients were HBsAg-positive. One of 403 HBsAg-negative patients tested positive for HBV DNA in serum but not in PBMCs. Positive controls: six of 20 patients were under antiviral therapy and had negative HBV DNA in both serum and PBMCs. In 11 of 14 remaining patients, HBV DNA was detected in serum and in both serum and PBMCs in 3 patients. Negative controls: All 34 patients were anti-HBc-negative and HBV DNA-negative in both serum and PBMCs. In the long term, the only case of anti-HBc-negative OBI lost anti-HBs 5years after inclusion in the study and showed HBV reactivation with HBsAg re-seroconversion. We found nil prevalence of OBI in CHD patients and a very low prevalence (<1%) in KTxR suggesting that routine screening for HBV DNA is not required in CHD population in our region. However, in KTxR, pretransplant screening with HBV DNA should be considered. Testing for HBV DNA in PBMCs does not seem to be of additional value.

Update Treatment for HBV Infection and Persistent Risk for Hepatocellular Carcinoma: Prospect for an HBV Cure.

Since the discovery of the hepatitis B virus (HBV) by Blumberg et al. in 1965, its genome, sequence, epidemiology, and hepatocarcinogenesis have been elucidated. Globally, hepatitis B virus (HBV) is still responsible for the majority of hepatocellular carcinoma (HCC). HCC is the sixth-most common cancer in the world and the second-most common cancer death. The ultimate goal of treating HBV infection is the prevention of HCC. Fortunately, anti-HBV treatment with nucleos(t)ide analogues (NAs), which began with lamivudine in 1998, has resulted in remarkable improvements in the survival of patients with chronic hepatitis B and a reduced incidence of HCC. These results were documented with lamivudine, entecavir, and tenofovir. Nonetheless, as the duration of antiviral treatment increases, the risk for HCC still remains despite undetectable HBV DNA in serum, as reported by different investigators with observation up to 4–5 years. In our own experience, we are witnessing the development of HCC in patients who have received antiviral treatment. Some have enjoyed negative serum HBV DNA for over 12 years before developing HCC. Current treatment with NAs can effectively suppress the replication of the virus but cannot eradicate the covalently closed circular DNA (cccDNA) that is within the nucleus of hepatocytes. There still remains a great need for a cure for HBV. Fortunately, several compounds have been identified that have the potential to eradicate HBV, and there are ongoing clinical trials in progress in their early stages.

Need For HBV Cure: Persistent Risk For Subsequent New And Recurrent HCC Even After A Decade Of Successful Anti-HBV Therapy And Initial Tumor Ablation

Since 1998 with the highly effective nucleos(t)ide analogues for Hepatitis B virus (HBV), we have witnessed successful viral suppression that has led to the delay or prevention of the development of hepatocellular carcinoma (HCC). Reduced incidence of HCC with anti-HBV treatment has been documented with lamivudine, entecavir and tenofovir. A number of investigators have observed that after the initial tumor was ablated, anti-HBV treatment could prevent or reduce the recurrent or subsequent new HCC. Nonetheless, as reported earlier, we are observing the persistent risk for HCC in patients in spite of successful viral suppression for over a decade with anti-HBV therapy. In this report, we present three cases who underwent initial tumor ablation and maintained negative HBV DNA with anti-HBV therapy and yet developed subsequent new or recurrent HCC at the intervals of 5,10,11 and 15 years. These persistent risks for new and recurrent HCC are attributed to the incomplete control of HBV due to the presence of cccDNA in the host’s hepatocytes. Current antiviral treatment can achieve a functional cure (suppression of HBV replication) while a complete cure of eliminating the cccDNA has not been possible. Therefore, even though HBV is not actively replicating, cccDNA remains in the nucleus of the hepatocytes and continues hepatocarcinogenic processes including HBV and host DNA integration. There is a need for HBV cure drugs.

Persistent Loss of Hepatitis B Virus Markers in Serum without Cellular Immunity by Combination of Peginterferon and Entecavir Therapy in Humanized Mice.

Nucleot(s)ide analogues and peginterferon (PEG-IFN) treatment are the only approved therapies for chronic hepatitis B virus (HBV) infection. However, complete eradication of the virus, as indicated by persistent loss of hepatitis B surface antigen (HBsAg), is rare among treated patients. This is due to long-term persistence of the HBV genome in infected hepatocytes in the form of covalently closed circular DNA (cccDNA). In this study, we investigated whether administration of a large dose of a nucleoside analogue in combination with PEG-IFN can achieve long-term loss of HBsAg in human hepatocyte chimeric mice. Mice were treated with a high dose of entecavir and/or PEG-IFN for 6 weeks. High-dose combination therapy with both drugs resulted in persistently negative HBV DNA in serum. Although small amounts of HBV DNA and cccDNA (0.1 and 0.01 copy/cell, respectively) remained in the mouse livers, some of the mice remained persistently negative for serum HBV DNA at 13 weeks after cessation of the therapy. Serum HBsAg and hepatitis B core-related antigen (HBcrAg) continued to decrease and eventually became negative at 12 weeks after cessation of the therapy. Analysis of the HBV genome in treated mice showed accumulation of G-to-A hypermutation and CpG III island methylation. Persistent loss of serum HBV DNA and loss of HBV markers by high-dose entecavir and PEG-IFN combination treatment in chimeric mice suggests that control of HBV can be achieved even in the absence of a cellular immune response.

Effect of entecavir in the treatment of patients with hepatitis B virus-related compensated and decompensated cirrhosis.

Chronic hepatitis B virus (CHB) infection is a burden on global healthcare and is associated with a higher risk of serious sequelae, including cirrhosis and hepatocellular carcinoma. The clinical application of entecavir as a treatment for CHB has produced positive outcomes, and so is an attractive form of pharmacological therapy. However, little data exists comparing the safety and efficacy of entecavir for the treatment of hepatitis B virus (HBV)-related compensated, and decompensated cirrhosis, respectively. The aim of the present study was to evaluate entecavir therapy as a treatment for patients with HBV-related compensated and decompensated cirrhosis. A retrospective analysis of 46 compensated patients (compensated group) and 51 decompensated cirrhotic patients (decompensated group) treated with entecavir was conducted. Baseline demographics, clinical outcomes, and adverse events during the treatment were compared. Treatment with entecavir for 96 weeks resulted in significant improvements in serum levels of HBV DNA (P=0.002), albumin (P=0.014), cholinesterase (CHE; P=0.001), HBV DNA negativity rate (P=0.004), Child-Turcotte-Pugh score (P=0.030), alanine aminotransferase normalized rate (P=0.039), and the degree of esophageal varices liver stiffness (P=0.002) in the two groups. However, statistical analysis revealed that the improvements were significantly higher in the compensated group compared with the decompensated group (P<0.05). The complement component (C)3 and C4 levels were also significantly increased in the compensated group compared with the decompensated group at weeks 24, 48 and 96 (P<0.05). In addition, the incidences of hepatocellular carcinoma, upper digestive tract hemorrhage and ascites were significantly higher in the decompensated group compared with the compensated group (P<0.05). In conclusion, treatment with 96-week entecavir therapy produced similar clinical outcomes in compensated and decompensated cirrhotic patients via inhibiting HBV-DNA viral load and recovering complement C3 and C4; however, entecavir exerts a better effect on patients with compensated cirrhosis, and so this therapy may improve the prognosis of such patients.

Effect on HBs antigen clearance of addition of pegylated interferon alfa-2a to nucleos(t)ide analogue therapy versus nucleos(t)ide analogue therapy alone in patients with HBe antigen-negative chronic hepatitis B and sustained undetectable plasma hepatitis B virus DNA: a randomised, controlled, open-label trial

Findings from uncontrolled studies suggest that addition of pegylated interferon in patients with HBe antigen (HBeAg)-negative chronic hepatitis B receiving nucleos(t)ide analogues with undetectable plasma hepatitis B virus (HBV) DNA might increase HBs antigen (HBsAg) clearance. We aimed to assess this strategy. In this randomised, controlled, open-label trial, we enrolled patients aged 18-75 years with HBeAg-negative chronic hepatitis B and documented negative HBV DNA while on stable nucleos(t)ide analogue regimens for at least 1 year from 30 hepatology tertiary care wards in France. Patients had to have an alanine aminotransferase concentration of less than or equal to five times the upper normal range, no hepatocellular carcinoma, and a serum α fetoprotein concentration of less than 50 ng/mL, normal dilated fundus oculi examination, and a negative pregnancy test in women. Patients with contraindications to pegylated interferon were not eligible. A centralised randomisation used computer-generated lists of random permuted blocks of four with stratification by HBsAg titres (< or ≥2·25 log10 IU/mL) to allocate patients (1:1) to receive a 48 week course of subcutaneous injections of 180 μg per week of pegylated interferon alfa-2a in addition to the nucleos(t)ide analogue regimen or to continue to receive nucleos(t)ide analogues only. The primary endpoint was HBsAg loss at week 96 by intention-to-treat analysis. This trial is closed and registered with ClinicalTrials.gov, number NCT01172392. Between Jan 20, 2011, and July 18, 2012, we randomly allocated 185 patients (92 [50%] to pegylated interferon and nucleos(t)ide analogues and 93 [50%] to nucleos(t)ide analogues alone). We excluded two patients from the pegylated interferon plus nucleos(t)ide analogues group from analyses because of withdrawal of consent (one patient) or violation of inclusion criteria (one patient). At week 96, loss of HBsAg was reported in seven (7·8%) of 90 patients in the pegylated interferon plus nucleos(t)ide analogues group versus three (3·2%) of 93 in the nucleos(t)ide analogues-alone group (difference 4·6% [95% CI -2·6 to 12·5]; p=0·15). 85 (94%) of 90 patients started pegylated interferon, three (4%) of whom had a dose reduction and 17 (20%) had an early discontinuation of pegylated interferon (seven [41%] for serious adverse events). Grade 3 and 4 adverse events were more frequent in the pegylated interferon plus nucleos(t)ide analogues group (26 [29%] grade 3 adverse events; 19 [21%] grade 4 adverse events) than in the nucleos(t)ide analogues-alone group (three [3%] grade 3; six [6%] grade 4). Addition of a 48 week course of pegylated interferon to nucleos(t)ide analogue therapy in patients with HBeAg-negative chronic hepatitis B with undetectable HBV DNA for a least 1 year was poorly tolerated and did not result in a significant increase of HBsAg clearance. Institut national de la santé et de la recherche médicale-Agence nationale de recherches sur le sida et les hépatites virales (France Recherche Nord&sud Sida-vih Hepatites).

Entecavir to Telbivudine Switch Therapy in Entecavir-Treated Patients with Undetectable Hepatitis B Viral DNA.

PurposeThis study examined 2-year outcome of consecutive therapy using entecavir (ETV) followed by telbivudine (LdT) in subjects with undetectable hepatitis B virus (HBV) DNA level and normal alanine aminotransferase level after the initial 6 months of ETV treatment.Materials and MethodsSixty subjects were randomized to continue with ETV or switch to LdT. Significant difference in baseline characteristics was not found between the two groups. Persistent HBV DNA level of 20–60 IU/mL in three consecutive samples collected three months apart or singly measured HBV DNA level of >60 IU/mL was defined as virological rebound.ResultsDuring 96 weeks of follow-up, all subjects of the ETV-only group (n=30) resulted in undetectable HBV DNA level. On the other hand, 83.3% (n=25) of the LdT-switched group showed treatment success. Virological rebound time varied from week 24 to 84 after switching to LdT. HBV DNA level was 180 to 2940 IU/mL at rebound time. All subjects with virological rebound (n=5) showed drug-resistant mutation: three had mutation rtM204I, and two had mutation rtM204V. Consecutive treatment using ETV followed by LdT showed virological rebound in 16.7% of subjects during 96 weeks of follow-up. HBV DNA negativity during initial ETV therapy could not be achieved in patients who switched to LdT.ConclusionConsecutive treatment using ETV followed by lamivudine was ineffective for treating chronic hepatitis B. LdT was found as a more potent antiviral agent than lamivudine. However, this conclusion requires larger-scale, long-term prospective reviews of the treatment effects of ETV-LdT switch therapy.

Lamivudine switch therapy in chronic hepatitis B patients achieving undetectable hepatitis B virus DNA after 3 years of entecavir therapy: A prospective, open-label, multicenter study

The subsequent maintenance therapy in chronic hepatitis B (CHB) patients after long-term viral replication suppression is still uncertain. We aim to evaluate the efficacy of lamivudine (LAM) maintenance therapy in CHB patients achieving undetectable hepatitis B virus (HBV) DNA after 3 years of entecavir (ETV) therapy. Consecutive CHB patients who received at least 3 years of ETV and achieved HBV DNA negativity were allocated either LAM switch therapy or stopped ETV therapy in a prospective, open-label study. Another group of sex- and age-matched patients with continuous ETV therapy for at least 4 years served as historical control group. The primary outcome measurement of the study was relapse of HBV DNA (defined as serum HBV DNA level ≥ 2000 IU/mL). A total of 74 patients, including 42 of LAM switch and 32 of the nonswitch group, were enrolled. There were no significant differences in demographics, except a higher proportion of patients with positive hepatitis B envelope antigen in the nonswitch group at the initiation of ETV therapy. The LAM switch group had significantly lower 1-year relapse rate of HBV within 1 year compared to the nonswitch group (14.3% vs. 75%, p<0.001). However, none of the 48 historical control patients developed relapse of HBV, which was significantly lower than the rate in LAM switch group (p < 0.001). LAM switch was the only factor associated with HBV DNA relapse. In conclusion, continuous long-term potent nucleot(s)ide analogue therapy is mandatory for prevention of viral relapse in CHB patients.