HBV DNA Levels Research Articles

CTLA4+CD4+CXCR5−FOXP3+ T cells associate with unfavorable outcome in patients with chronic HBV infection

BackgroundA major barrier to achieving a favorable outcome of chronic HBV infection is a dysregulated HBV-specific immune response resulting from immunosuppressive features of FOXP3+ T cells. A better definition of FOXP3+ T cells is essential for improving the prognosis of HBV infection. We aimed to investigate the role of CD4+CXCR5−FOXP3+ T cells with CTLA4 expression in patients with chronic HBV infection.MethodsTreatment-naïve chronic HBV-infected patients, HBV-related hepatic failure, and a longitudinal cohort of chronic hepatitis B (CHB) patients with nucleos(t)ide analogue treatment were enrolled for analysis of CD4+CXCR5−FOXP3+ T cell responses by flow cytometry and single-cell RNA sequencing (scRNA-seq).ResultsScRNA-seq revealed that circulating CD4+CXCR5−FOXP3+ T cells presented distinct inhibitory features compared to spleen tissue. Meanwhile, patients with treatment-naïve chronic HBV infection or with HBV-related hepatic failure showed an upregulation of immune-suppressive features (PD-1, CTLA4, GITR) on CD4+CXCR5−FOXP3+T cells; in vitro analysis found HBeAg and HBcAg stimulation induced elevated levels of inhibitory molecules. Notably, the frequency of CTLA4+CD4+CXCR5−FOXP3+ T cells was positively correlated with HBV DNA levels, and longitudinal analysis demonstrated a high frequency of this subset at 12 weeks of antiviral treatment predicted unfavorable outcome in CHB patients.ConclusionsCTLA4+CD4+CXCR5−FOXP3+ T cells are related to unfavorable outcomes in HBV-infected patients; these data indicated that alleviating CTLA4+CD4+CXCR5−FOXP3+ T cells may improve the prognosis of HBV infection.

Ascites re-compensation in HBV-related first decompensated cirrhosis after anti-viral therapy.

Effective antiviral therapy can significantly improve the long-term prognosis of HBV-related decompensated patients, and re-compensation may be achieved in part of the patients. To explore the re-compensation of ascites after HBV suppression and the risk factors, the clinical outcomes of 196 consecutive patients with HBV-related first decompensated cirrhosis of ascites treated with nucleos(t)ide analogue (NUC) were analyzed retrospectively. Among these patients, the median serum HBV DNA level was 5.0 (IQR, 3.0-6.0) log10 IU/mL before treatment. Most patients were given NUC with high barrier to resistance including ETV (152), TDF (1) and TAF (1). Initial combination of LAM plus ADV and LdT plus ADV was used in 41 patients and 1 patients, respectively. After NUC treatment, the percentage of patients with ascites regression was 77.6%, 81.4%, 70.5%, 93.8%, 80.8% at 12, 24, 36, 48, 60 months, respectively (P<0.001). The distribution of ascites severity showed that the patients' ascites improved, with the proportion of no ascites and mild ascites gradually increased. The proportion of re-compensation of ascites defined as negative HBV DNA, improved liver function and ascites regression (off diuretics) was 59.7%, 70.0%, 52.3%, 59.4%, 46.2% at 12, 24, 36, 48, 60 months (P<0.001). The rate of ascites regression was higher in viral response (VR) cohort when compared with that in non-VR cohort. Univariate and multivariable analysis showed that level of serum ALT (OR:0.988, 95%CI, p=0.029) and load of serum HBV DNA (OR:0.78895%CI, p=0.044) at baseline were risk factors of re-compensation of ascites. This study demonstrated that antiviral therapy could reverse decompensation of ascites in HBV-related first decompensated cirrhosis and the level of ALT and HBV DNA were risk factors of ascites re-compensation.

Effective Analysis of Antiviral Treatment in Patients with HBeAg-Seropositive Chronic Hepatitis B with ALT < 2 Upper Limits of Normal: A Multi-center Retrospective Cohort Study.

Although the indications for antiviral therapy for patients with chronic hepatitis B have been gradually expanded in different guidelines, antiviral treatment efficacy remains unclear among HBeAg-seropositive patients with alanine aminotransferase (ALT) < 2 upper limits of normal (ULN). This study aimed to evaluate the efficacy of antiviral therapy for these patients. In total, 102 treatment-naive patients who were HBeAg seropositive with ALT < 2 ULN and had received nucleotide analogs were included, and their clinical data were retrospectively analyzed. After 96-week treatment, 84.3% (n = 86), 26.5% (n = 27) and 20.6% (n = 21) patients achieved virological response, HBeAg seroclearance and HBeAg seroconversion, respectively. Logistic regression analysis revealed that baseline AST (odds ratio [OR] = 1.069, 95% confidence interval [CI] 1.014-1.127, p = 0.014), serum HBV DNA (OR = 0.540, 95% CI 0.309-0.946, p = 0.031) and quantitative HBsAg levels (OR = 0.147, 95% CI 0.036-0.597, p = 0.007) were independent factors for virological response. At baseline, HBsAg < 4.63 log10 IU/ml was identified as a strong predictor for the 96-week virological response, with a concordance rate of 0.902. Moreover, the levels of liver stiffness values (8.30 ± 3.86 vs. 6.17 ± 1.91, p < 0.001) at week 96 had significantly declined compared to baseline. Nucleotide analog treatment effectively suppressed HBV DNA in patients with HBeAg-seropositive chronic hepatitis B with ALT < 2 × ULN and greatly improved liver fibrosis. The study also found that HBsAg < 4.63 log10 IU/ml was a strong predictor of the virological response.

Profiling of Peripheral TRBV and CD4+CD25+ Treg in CHB Patients with HBeAg SC during TDF Treatment.

It is lacking that markers could predict the prognosis of chronic hepatitis B (CHB) subjects during antiviral treatment, and the related cellular immune mechanism is not fully evaluated. To explore the comprehensive profile of T cell receptor β-chain (TRBV) and CD4+CD25+ regulatory T cell (Treg) in peripheral blood of CHB patients with HBeAg seroconverting (SC) during tenofovir disoproxil fumarate (TDF) treatment. The frequency of CD4+CD25high+ Treg and number of skewed TRBV in 20 HBeAg positive patients were determined at baseline and following every 12 weeks during 96-week TDF treatment. The relationship among serum alanine aminotransferase (ALT) level, HBV DNA load, Treg frequency, and the number of skewed TRBV, respectively, was analyzed for CHB patients. Receiver operative characteristic curve was applied to analyze their diagnostic value for HBeAg SC. The number of skewed TRBV at week 48, Treg frequency at week 72, and ALT level at baseline could predict the HBeAg SC or non-SC in CHB patients during 96-week TDF treatment. Moreover, the positive correlation between ALT or HBV DNA and Treg levels or skewed TRBVs was significant in the SC group, but not in non-SC. The predictive cutoff value of ALT for HBeAg SC was 178 U/L at baseline. Moreover, the ALT, Treg, and TRBV families would be associated with the prognosis and pathogenesis of CHB patients during TDF treatment.

Importance of CHB's grey zone: analysis of patients with HBeAg negative chronic hepatitis B virus infection.

HBeAg-negative chronic HBV infection is defined by viremia < 2,000 IU/ml (or < 20,000 IU/ml), normal ALT activity and minimal liver fibrosis. Some patients do not meet all the criteria and belong to the so-called grey zone. The aim of the work was to analyse a group of patients with asymptomatic chronic HBV infection, divide them according to the levels of HBV DNA during follow-up and to compare the clinical and laboratory parameters of the patients within the groups. We retrospectively analysed patients with HBeAg-negative chronic HBV infection examined in the Centre for Viral Hepatitis of the Department of Infectology in Košice, Slovakia, from September 2018 to December 2021. Patients were divided into three groups based on HBV DNA levels ​​during long-term follow-up ( 2,000 IU/ ml). We evaluated selected demographic, anamnestic and laboratory data (HBV DNA, ALT, fibrosis stage). Of the 280 enrolled patients, 160 were men (57.1 %), the average age was 48.0 years, and the mean length of follow-up was 4.7 years. HBV DNA levels ​​were consistently 2,000 IU/ml in 62 patients. 165 patients had normal ALT activity, 74 had fluctuating ALT activity, and permanently increased ALT in 41 patients. 139 patients underwent transient elastography examination, 16 of them had stage F2 fibrosis, two stage F3 and 1 had cirrhosis. When comparing the three groups divided according to HBV DNA, patients with fluctuating HBV DNA had the longest follow-up, but patients with HBV DNA permanently over 2,000 IU/ml were the youngest and the highest proportion of them had elevated ALT activity. 165 patients (58.9%) met the extended criteria of asymptomatic carriers, 115 were in the grey zone. Patients with HBeAg-negative chronic HBV infection often have fluctuating HBV DNA and ALT values ​​during follow-ups. Statistically significantly higher proportion of abnormal ALT activity in patients with HBV DNA > 2,000 IU/ml may suggest higher risk of adverse outcomes. Initiation of treatment in such patients is not always necessary unless they also meet the other indication criteria for treatment. The exact definition of the grey zone is currently absent (Tab. 2, Fig. 2, Ref. 16).

A Review on the Epidemiology of HBV and HIV Co-Infection

HIV infection has an impact on the natural course of chronic HBV infection, as it results in an increase in HBV DNA levels, accelerated progression of liver disease, and increased liver-related mortality compared with HBV mono-infection. In HIV/HBV co-infected individuals, widespread uptake and early initiation of HBV-active antiretroviral therapy have drastically improved the natural history of viral infection; however, liver disease remains common. In this paper we have reviewed the epidemiology, prevalence, treatment, pathogenesis of HIV and HBV co-infection. Co-infection of HBV-HIV is more likely to progress and complicate the disease and should be closely monitored and treated.

Correlation between serum quantitative HBsAg and HBV DNA levels in chronic hepatitis B patients

Background/Aim. Quantitative hepatitis B virus (HBV) surface antigen (qHBsAg) has become increasingly wide-spread in the last few years in both diagnostic and therapeutic protocols for HBV infection. Numerous studies have proposed it as a surrogate marker for covalently closed circular DNA (cccDNA). The aim of the study was to determine the correlation between qHBsAg and HBV DNA viremia in untreated patients. Methods. The study included 112 untreated patients diagnosed with chronic HBV infection. Demographic and other data from medical records and laboratory analyses, taken as part of routine chronic HBV infection diagnosis with the determination of qHBsAg and HBV DNA viremia, were recorded for all patients. Results. The average age of the patients included in the study was 48.27 ? 15.14 years; males (58%) were more represented. qHBsAg levels had a high-intensity positive correlation with HBV DNA viremia. The concentration of qHBsAg, HBV DNA viremia, and the concentrations of alanine aminotransferase and aspartate aminotransferase showed statistically significantly higher values in HBV e antigen (HBeAg)-positive than in HBeAg-negative patients. Conclusion. Our study showed that qHBsAg has a high-intensity positive correlation with HBV DNA viremia. The use of qHBsAg is essential for determining the phase of chronic HBV infection, assessment of the success and length of treatment, as well as for safe discontinuation of antiviral therapy with a lower risk of relapse.

Values of Serum HBsAg and HBeAg Levels for Virological Response of Patients with HBV-Related Liver Cirrhosis Treated by Entecavir.

The aim is to explore the clinical values of quantitative detection of serum hepatitis B surface antigen (HBsAg) and hepatitis B virus e antigen (HBeAg) levels for the virological response [hepatitis B virus (HBV)-deoxyribonucleic acid (DNA) level] of patients with HBV-related liver cirrhosis (HBV-LC) treated by entecavir. A total of 147 patients with HBV-LC treated from January 2016 to January 2019 were assigned to virol-ogical response (VR) group (n = 87) and no virological response (NVR) group (n = 60) according to whether virological response occurred after treatment. The values of serum HBsAg and HBeAg levels for predicting virological response were determined by receiver operating characteristic (ROC) curve, Kaplan-Meier survival analysis, and 36-Item Short Form Survey (SF-36). Serum HBsAg and HBeAg levels before treatment were positively correlated with HBV-DNA level in patients with HBV-LC, and there were significant differences in serum HBsAg and HBeAg levels in the 8th, 12th, 24th, 36th, and 48th weeks of treatment (p < 0.01). In the 48th week of treatment, the area under the ROC curve (AUC) of serum HBsAg log value for predicting virological response was the largest [0.818, 95% confidence interval (CI): 0.709 - 0.965], and the optimal cutoff value, sensitivity, and specificity was 2.53 ± 0.53 IU/mL, 91.34%, and 71.93%, respectively. AUC of serum HBeAg level for predicting virological response was the largest (0.801, 95% CI: 0.673 - 0.979), and the optimal cutoff value, sensitivity, and specificity was 27.38 ± 2.14 PEIU/mL, 88.52%, and 83.42%, respectively. Serum HBsAg and HBeAg levels are correlated with the virological response of patients with HBV-LC treated by entecavir.

Increased Non-MAIT CD161+CD8+ T Cells Display Pathogenic Potential in Chronic HBV Infection

CD161-expressing CD8+ T cells consist of mucosal-associated invariant T cells with semi-invariant T-cell receptor (TCR) use and non-mucosal-associated invariant T CD161+CD8+ T cells with polyclonal TCR repertoire. Although CD161+CD8+ T cells are enriched in liver and embrace hepatitis B virus (HBV)-specific T cells in chronic hepatitis B (CHB) patients, their roles in disease progression remain poorly understood. This study aimed to decipher their profiling and dynamic changes during chronic HBV infection. Blood samples from 257 CHB patients and nontumor liver specimens from 73 HBV-positive patients were analyzed for CD161+CD8+ T-cell characterization by flow cytometry, TCR repertoire determination, transcriptomic analyses, and cell experiments. CD161+CD8+ T cells were increased and hyperactivated in patients, while positive correlation between the CD161+CD8+ T-cell ratio and HBV-DNA level suggested this was insufficient to control HBV replication. The overlap of complementarity determining region 3 sequences supported the switch between CD161-CD8+ and CD161+CD8+ populations. Although CD161+CD8+ T cells were endowed with innateness phenotype and enhanced antiviral capacity, the population from patients had impaired type I cytokine production, and increased interleukin 17 and granzyme B secretion. The increased CD161+CD8+ T cells and their increased granzyme B secretion correlated positively with inflammation-associated liver injury. Hepatic CD161+CD8+ T cells showed neutrophil-related pathogenic potential because they had increased transcript signatures and proinflammatory cytokine production in neutrophil recruitment- and response-related pathways that changed consistently in the injured liver. Our results highlight the reduced antiviral potency but increased pathogenic potential of CD161+CD8+ T cells in CHB patients, supporting CD161 expression as a marker of pathogenic CD8+ T subset and the intervention target for liver injury.

A Correlation Study of Liver Biopsy Appearances, Serum HBV DNA and HBsAg Quantification Levels of Nigerian Patients with Chronic Liver Disease

Background: Infection with the hepatitis B virus is highly endemic in much of sub-Saharan Africa where it remains an important cause of chronic ill-health. Quantitative assay of HBV DNA and HBsAg are emerging as useful tools in the management of chronic HBV infection. However, it is not yet clear how well each of these might accurately correlate with the extent of liver damage during the course of the disease. Nevertheless, liver biopsy, albeit an invasive test, affords a superior direct assessment of all stages of the disease. Objective: Thus, we set out to determine how well-correlated serum levels of HBV-DNA and/or HBsAg might be with each other and with the liver biopsy appearances. Methodology: This study was prospective and we enrolled, 83 new treatment-naïve patients, who met the study criteria. Each patient had liver biopsy assessments of the grade of necroinflammation (A) and stage of fibrosis (F), according to the METAVIR Scoring System. In addition, quantitative HBsAg serum levels by immunoassay and HBV DNA by real-time quantitative PCR protocols were made. The Spearman’s and Kruskal-Wallis statistical tests were employed to determine the correlations between these assays separately and with the respective histological grades and stages of liver biopsy. (p=0.05) Results: The greatest number of the patients were within the 21-40 year- age group. Using the Spearman rho’s statistical correlation test, we found a positive but weak correlation between the serum HBsAg and HBVDNA levels (p=0.075, r=0.198). The Kruskal-Wallis analysis revealed statistically significant correlation between HBV-DNA and the grade of necroinflammation but not with the stage of fibrosis. Conclusions: We found that the serum HBV DNA levels had the only statistically significant correlation with the liver histological disease. This relationship needs to be further examined in larger studies in the future.

Discovery of a first-in-class orally available HBV cccDNA inhibitor

The persistence of covalently closed circular DNA (cccDNA) in infected hepatocytes is the major barrier preventing viral eradication with existing therapies in patients with chronic hepatitis B. Therapeutic agents that can eliminate cccDNA are urgently needed to achieve viral eradication and thus HBV cure. A phenotypic assay with HBV-infected primary human hepatocytes (PHHs) was employed to screen for novel cccDNA inhibitors. A HBVcircle mouse model and a uPA-SCID (urokinase-type plasminogen activator-severe combined immunodeficiency) humanized liver mouse model were used to evaluate the anti-HBV efficacy of the discovered cccDNA inhibitors. Potent and dose-dependent reductions in extracellular HBV DNA, HBsAg, and HBeAg levels were achieved upon the initiation of ccc_R08 treatment two days after the HBV infection of PHHs. More importantly, the level of cccDNA was specifically reduced by ccc_R08, while it did not obviously affect mitochondrial DNA. Additionally, ccc_R08 showed no significant cytotoxicity in PHHs or in multiple proliferating cell lines. The twice daily oral administration of ccc_R08 to HBVcircle model mice, which contained surrogate cccDNA molecules, significantly decreased the serum levels of HBV DNA and antigens, and these effects were sustained during the off-treatment follow-up period. Moreover, at the end of follow-up, the levels of surrogate cccDNA molecules in the livers of ccc_R08-treated HBVcircle mice were reduced to below the lower limit of quantification. We have discovered a small-molecule cccDNA inhibitor that reduces HBV cccDNA levels. cccDNA inhibitors potentially represent a new approach to completely cure patients chronically infected with HBV. Covalently closed circular DNA (cccDNA) persistence in HBV-infected hepatocytes is the root cause of chronic hepatitis B. We discovered a novel small-molecule cccDNA inhibitor that can specifically reduce cccDNA levels in HBV-infected hepatocytes. This type of molecule could offer a new approach to completely cure patients chronically infected with HBV.

Intrahepatic quantification of HBV antigens in chronic hepatitis B reveals heterogeneity and treatment-mediated reductions in HBV core-positive cells

Patterns of liver HBV antigen expression have been described but not quantified at single-cell resolution. We applied quantitative techniques to liver biopsies from individuals with chronic hepatitis B and evaluated sampling heterogeneity, effects of disease stage, and nucleos(t)ide (NUC) treatment, and correlations between liver and peripheral viral biomarkers. Hepatocytes positive for HBV core and HBsAg were quantified using a novel four-plex immunofluorescence assay and image analysis. Biopsies were analysed from HBeAg-positive (n= 39) and HBeAg-negative (n= 75) participants before and after NUC treatment. To evaluate sampling effects, duplicate biopsies collected at the same time point were compared. Serum or plasma samples were evaluated for levels of HBV DNA, HBsAg, hepatitis B core-related antigen (HBcrAg), and HBV RNA. Diffusely distributed individual HBV core+ cells and foci of HBsAg+ cells were the most common staining patterns. Hepatocytes positive for both HBV core and HBsAg were rare. Paired biopsies revealed large local variation in HBV staining within participants, which was confirmed in a large liver resection. NUC treatment was associated with a >100-fold lower median frequency of HBV core+ cells in HBeAg-positive and HBeAg-negative participants, whereas reductions in HBsAg+ cells were not statistically significant. The frequency of HBV core+ hepatocytes was lower in HBeAg-negative participants than in HBeAg-positive participants at all time points evaluated. Total HBV+ hepatocyte burden correlated with HBcrAg, HBV DNA, and HBV RNA only in baseline HBeAg-positive samples. Reductions in HBV core+ hepatocytes were associated with HBeAg-negative status and NUC treatment. Variation in HBV positivity within individual livers was extensive. Correlations between the liver and the periphery were found only between biomarkers likely indicative of cccDNA (HBV core+ and HBcrAg, HBV DNA, and RNA). HBV infects liver hepatocyte cells, and its genome can exist in two forms that express different sets of viral proteins: a circular genome called cccDNA that can express all viral proteins, including the HBV core and HBsAg proteins, or a linear fragment that inserts into the host genome typically to express HBsAg, but not HBV core. We used new techniques to determine the percentage of hepatocytes expressing the HBV core and HBsAg proteins in a large set of liver biopsies. We find that abundance and patterns of expression differ across patient groups and even within a single liver and that NUC treatment greatly reduces the number of core-expressing hepatocytes.

Factors affecting HBV DNA suppression in chronic hepatitis B patients treated with tenofovir disoproxil fumarate

Background: This study aims to determine the factors affecting HBV DNA suppression in chronic hepatitis B patients with tenofovir disoproxil fumarate (TDF). Methods: A case-control was carried out from October 2021 to August 2022 on 182 chronic hepatitis B patients who had TDF therapy regularly for 24 weeks at H. Adam Malik and USU Hospitals in Medan, Indonesia. The history of the samples was obtained, followed by physical examination, and blood collection. CTLA-4 polymorphism examination was carried out using real-time PCR, while the serum CTLA-4 levels were assessed with ELISA. Results: The results showed that CTLA-4, HBV DNA, ALT, and CTLA-4 -1661G&gt;A polymorphisms have a relationship with HBV DNA suppression in chronic hepatitis B patients with TDF. Conclusions: The levels of CTLA-4, HBV DNA, ALT, and CTLA-4 -1661G&gt;A polymorphism have a potential relationship with the suppression of HBV DNA in chronic hepatitis B patients with TDF.

Levels of HBV RNA in chronic HBV infected patients during first-line nucleos(t)ide analogues therapy

BackgroundSerum HBV RNA has been considered a potential biomarker in monitoring the prognosis of chronic hepatitis B (CHB). However, Real-life cohort studies on the profile of HBV RNA in chronic HBV infected patients during first-line nucleos(t)ide analogues (NAs) are lacking. We aimed to investigate HBV RNA dynamic pattern and clinical value chronic HBV infected patients under NA therapy.MethodsHBV RNA and clinical assessments were measured in 82 treatment-naïve chronic HBV infected patients. These enrolled patients were categorized into HBeAg-positive chronic HBV infected (n = 53) and HBeAg-negative chronic HBV infected (n = 29). Of these, there were 59, 46, and 30 chronic HBV infected patients completed the follow-up clinical assessments at 12, 24, and 48 weeks of NAs therapy, respectively.ResultsIn treatment-naïve patients, there was a positive correlation between HBV RNA and HBV DNA, HBsAg (r = 0.602 and 0.502. P < 0.05). The median level of HBV DNA was higher than HBV RNA by 1.64 log10 copies/mL. The mean level of serum HBV RNA was 4.62 (IQR: 3.05–5.82) log10 copies/mL at baseline, and the median level of HBV RNA was 2.88 (IQR: 0–4.67), 2.71 (IQR: 0–4.22), and 2.96 (IQR: 0–4.32) log10 copies/mL at week 12, 24, and 48, respectively. HBV RNA showed a positive linear correlation with HBV DNA at 12, 24, and 48 weeks of NA treatment (r = 0.640, 0.715, and 0.656 respectively, P < 0.05). In patients who were treated 48 weeks NAs, 67% had quantifiable HBV RNA while only 37% had quantifiable HBV DNA.ConclusionHBV RNA has signature profiles in different stages of chronic HBV infected patients receiving first-line NAs. During antiviral treatment, HBV RNA can still monitor the virus activity in patients whose serum HBV DNA cannot be detected.

Direct-Acting Antivirals and the Risk of Hepatitis B Reactivation in Hepatitis B and C Co-Infected Patients: A Systematic Review and Meta-Analysis.

Hepatitis B (HBV) reactivation was observed to be more than 10% in patients receiving interferon-based therapy for hepatitis C (HCV) co-infection. At present, when direct-acting antiviral (DAA) has become the main treatment for HCV, there are few large-scale studies on the reactivation of HBV in these population. We studied HBV reactivation risk and prophylactic HBV treatment efficacy in HBV/HCV co-infected patients receiving DAA therapy. Relevant studies were selected from the Ovid-Medline, Ovid-EMBASE, Cochrane Central Register of Controlled Trials, KoreaMed, KMbase, and RISS databases through 4 September 2020. Data pooling was carried out using the random-effects method. We identified 39 articles with 119,484 patients with chronic (n = 1673) or resolved (n = 13,497) HBV infection under DAA therapy. When the studies were pooled, the HBV reactivation rate was 12% (95% confidence interval (CI) 6-19, I2 = 87%), indicating that this population needs careful attention. When stratified by baseline HBV DNA, the undetectable HBV DNA group showed a significantly lower risk of reactivation than the detectable HBV DNA group (odds ratio (OR) 0.30, 95% CI 0.11-0.86, I2 = 0%). Prophylactic HBV therapy reduced HBV reactivation risk (OR 0.25, 95% CI 0.07-0.92, I2 = 0%). Patients with a resolved HBV infection showed a negligible rate (0.4%) of HBV reactivation. In conclusion, patients with detectable HBV DNA levels warrant careful monitoring for HBV reactivation and may benefit from preventive anti-HBV treatment.

Functional molecular expression of nature killer cells correlated to HBsAg clearance in HBeAg-positive chronic hepatitis B patients during PEG-IFN α-2a therapy.

To explore whether the frequencies and functional molecules expression of Natural Killer cells (NK cells) are related to hepatitis B surface antigen (HBsAg) disappearance in hepatitis B e envelope antigen (HBeAg)-positive patients with chronic hepatitis B (CHB) throughout peginterferon alpha-2a (PEG-IFN α-2a) treatment. In this prospective research, HBeAg-positive patients with CHB received PEG-IFN α-2a treatment, completing 4-year follow-up. After PEG-IFN α-2a treatment, undetectable HBV DNA, HBsAg loss, and HBeAg disappearance were defined as functional cure. Proportions of NK, CD56dim, CD56bright, NKp46+, NKp46dim, NKp46high, and interferon alpha receptor 2 (IFNAR2)+ NK cells, and the mean fluorescence intensity (MFI) of NK cell surface receptors IFNAR2 and NKp46 were detected. 66 patients were enrolled into the study in which 17 patients obtained functional cure. At baseline, hepatitis B virus desoxyribose nucleic acid (HBV DNA) titer in patients with functional cure was remarkably lower than that in Non-functional cure group. Compared with baseline, HBV DNA levels, HBsAg levels, and HBeAg levels significantly declined at week 12 and 24 of therapy in patients with functional cure. At baseline, the negative correlation between CD56bright NK% and HBV DNA and the negative correlation between CD56dim NK% and HBV DNA was showed; CD56bright NK% and IFNAR2 MFI in patients with functional cure were remarkably higher than those in patients without functional cure. After therapy, CD56bright NK% and NKp46high NK% in patients with functional cure were higher than those in patients without functional cure. In Functional cure group, after 24 weeks of treatment NK%, CD56bright NK%, IFNAR2 MFI weakly increased, and NKp46high NK% and NKp46 MFI significantly increased, meanwhile, CD56dim NK% and NKp46dim NK% decreased. Only NKp46 MFI increased after therapy in patients without functional cure. The lower HBV DNA load and the higher CD56bright NK% before therapy, and the higher the post-treatment CD56bright NK%, IFNAR2 MFI, NKp46high NK%, the easier to achieve functional cure.

Anti-hepatitis B virus activity of lithospermic acid, a polyphenol from Salvia miltiorrhiza, in vitro and in vivo by autophagy regulation

Ethnopharmacological relevanceSalvia miltiorrhiza (the roots of S. miltiorrhiza Bunge, Danshen in Chinese), a traditional Chinese medicine, has been clinically used to prevent and treat various diseases, such as cardiovascular and cerebrovascular diseases, diabetes, and hepatitis B, in China and some other Asian countries. Lithospermic acid (LA), a polyphenol derived from S. miltiorrhiza, has been reported to exhibit multiple pharmacological properties, such as anti-inflammatory, anti-HIV, and anti-carbon tetrachloride-induced liver injury activities. However, little is known about the anti-hepatitis B virus (HBV) activity of LA. Aim of the studyThe study was projected to investigate the anti-HBV activity of LA in vitro (HepG2.2.15 and pHBV1.3-transfected HepG2 cells) and in vivo (pAAV-HBV1.2 hydrodynamic injection [HBV-HDI] mice) and explore the potential mechanism as well. Materials and methodsHepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) contents were detected by ELISA kits. HBV DNA and hepatitis B core antigen (HBcAg) levels were evaluated by quantitative real-time polymerase chain reaction and immunohistochemistry assay, respectively. The proteins in autophagy process, lysosomal acidic function, and autophagy-related signaling pathways were examined by Western blot. Transmission electron microscopy was used to observe the number of autophagosomes and autolysosomes. Confocal microscopy was applied to analyze the autophagic flux and lysosomal acidification, using mCherry-enhanced green fluorescent protein (EGFP)-microtubule-associated protein light chain (LC)3 and lysosomal probes, respectively. ResultsLA exhibited anti-HBV activity by inhibiting HBV DNA replication in HepG2.2.15 and pHBV-transfected HepG2 cells in dose- and time-dependent manners and hampering HBsAg and HBeAg levels in HepG2.2.15 cells to a certain extent. LA reduced HBV DNA, HBsAg/HBeAg, and HBcAg levels in the serum/liver tissues of HBV-HDI C57BL/6 mice during the 3-week treatment and suppressed the withdrawal rebound of HBV DNA and HBsAg in the mice serum. LA increased LC3-II protein expression and the number of autolysosomes/autophagosomes and promoted the degradation of sequestosome 1(p62) protein in vitro and in vivo. LA enhanced the co-localization of LC3 protein with autolysosomes, further confirming the ability of LA to induce a complete autophagy. Knockdown of autophagy-related gene (Atg) 7 or 5 in vitro and administration of 3-methyladenine (an autophagic inhibitor) in vivo disabled the inhibitory efficacy of LA on HBV DNA replication, suggesting that the anti-HBV efficacy of LA depended on its ability of inducing autophagy. LA could enhance lysosomal acidification and improve the function of lysosomes by promoting the protein expression of lysosomal-associated membrane protein (LAMP)-1, LAMP-2, and mature cathepsin D, which may contribute to the autophagic induction of LA. LA inhibited the activation of AKT and mammalian target of rapamycin (mTOR) induced by HBV, which was reversed by IGF-1 (an agonist of the PI3K/AKT/mTOR signaling pathway), indicating that LA elicited autophagy through hampering the PI3K/AKT/mTOR signaling pathway. ConclusionWe revealed the anti-HBV activity and mechanism of LA in vitro and in vivo. This study facilitates a new understanding of the anti-HBV potent components of S. miltiorrhiza and sheds light on LA for further development as an active constituent or candidate used in the therapy against HBV infection.

96-Week Treatment of Tenofovir Amibufenamide and Tenofovir Disoproxil Fumarate in Chronic Hepatitis B Patients.

Tenofovir amibufenamide (TMF) is a novel phosphoramidated prodrug of tenofovir with noninferior efficacy and better bone and renal safety to tenofovir disoproxil fumarate (TDF) in 48 weeks of treatment. Here, we update 96-week comparison results. Patients with chronic hepatitis B were assigned (2:1) to receive either 25 mg TMF or 300 mg TDF with matching placebo for 96 weeks. The virological suppression was defined as HBV DNA levels <20 IU/mL at week 96. Safety was evaluated thoroughly with focusing on bone, renal, and metabolic parameters. Virological suppression rates at week 96 were similar between TMF and TDF group in both HBeAg-positive and HBeAg-negative populations. Noninferior efficacy was maintained in the pooled population, while it was first achieved in patients with HBV DNA ≥7 or 8 log10 IU/mL at baseline. Non-indexed estimated glomerular filtration rate for renal safety assessment was adopted, while a smaller decline of which was seen in the TMF group than in the TDF group (p=0.01). For bone mineral density, patients receiving TMF displayed significantly lower reduction levels in the densities of spine, hip, and femur neck at week 96 than those receiving TDF. In addition, the lipid parameters were stable after week 48 in all groups while weight change still showed the opposite trend. TMF maintained similar efficacy at week 96 compared with TDF with continued superior bone and renal safety profiles (NCT03903796).

Sequence characterization of extracellular HBV RNA in patient plasma.

Antiviral nucleos(t)ide analogue therapies inhibit HBV replication and suppress the HBV DNA levels in patients with chronic HBV infection. Since HBV RNAs are expressed from cccDNA or HBV integrated sequences, independently of viral genome replication, levels of HBV RNAs in plasma may remain high following treatment with nucleos(t)ide analogue. Thus, HBV RNAs have been proposed to be used as a viral biomarker for treatment outcome and disease progression. Recent investigations of plasma HBV RNAs described the presence of full length as well as subgenomic forms of RNA. To support the usage of plasma HBV RNAs as a viral biomarker, further understanding of HBV RNA composition in clinical samples is needed. Here, sequence of extracellular HBV RNAs was characterized in plasma samples of patients with chronic HBV infection using two independent RNA amplification methods that do not use HBV-specific primers for amplification: total RNA (NuGEN RNAseq) and mRNA (TruSeq RNAseq). Sequencing coverage was obtained across the full length of HBV genome for both methods, confirming the presence of full-length HBV RNA in plasma. The sequence of HBV RNA was nearly identical to plasma HBV DNA sequence in each sample with only 0-14 (median 4) mismatches over 3kb. Thus, sequence of HBV RNA plasma reflects the intrahepatic viral reservoir and can be used for monitoring of sequence variants such as resistance in clinical trials. Additionally, RNA splice forms, different polyA tails start positions and presence of HBV-human chimeric transcript were identified.

Effect of tenofovir disoproxil and telbivudine on the growth and development of infants by blocking mother-to-child transmission of hepatitis B virus

Purpose: To investigate the effect of tenofovir disoproxil and telbivudine on the growth and development of infants after blocking mother-to-child transmission (MTCT) of hepatitis B virus (HBV).&#x0D; Methods: Seventy pregnant women with chronic hepatitis B (CHB) were recruited and allocated to tenofovir disoproxil group (n = 35) and telbivudine group (n = 35) using random number table method. Tenofovir disoproxil group was given 300 mg tenofovir disoproxil orally four times daily, while telbivudine group was given telbivudine 600 mg orally four times daily.&#x0D; Results: After treatment, both groups showed no significant differences in serum HBV-DNA and ALT levels before delivery and 3 months after delivery (p &gt; 0.05). Both groups showed no remarkable differences in the incidence of hypohydramnios, cholestasis, hypothyroidism, anemia, prolonged labor, fetal distress, and placental adhesions (p &gt; 0.05). Both groups showed no significant differences in the rates of premature rupture of membranes, preterm birth, vaginal delivery, and cesarean section (p &gt; 0.05). Both groups showed no significant differences in neonatal sex, gestational age at birth, weight, length, and Apgar scores (p &gt; 0.05). The differences in the positive rates of HBVsAg, HBsAb, and HBeAg at birth and at 12 months were not statistically noticeable (p &gt; 0.05).&#x0D; Conclusion: Tenofovir disoproxil and telbivudine reduce HBV-DNA levels, effectively blocks MTCT, and have a similar safety profile for infants. Further investigations to confirm th