Interferon Gamma-Inducible Protein 16 of Peripheral Blood Mononuclear Cells May Sense Hepatitis B Virus Infection and Regulate the Antiviral Immunity

Interferon gamma-inducible protein 16 (IFI16) is a DNA sensor protein, which triggers interferon-beta (IFN-β) production. However, the role of IFI16 in the innate immunity against hepatitis B virus (HBV) remains controversial. Peripheral blood mononuclear cells (PBMCs) and serum specimens were collected from 20 patients with chronic hepatitis B (CHB) receiving Peg-IFN-α2b therapy. IFI16 mRNA/protein of PBMCs and serum IFI16 at baseline and changes during Peg-IFN-α2b treatment were detected. The interaction between IFI16 and HBV DNA in the PBMCs was analyzed using chromatin immunoprecipitation assay. Leukemic T cell line CEM-C7 and HBV-replicating HepG2.2.15 cells were used to test the effects of interferon treatment and HBV replication on IFI16 expression. Compared with healthy controls, lower levels of IFI16 mRNA but more significant expression of IFI16 protein with heterogeneous degradation were detected in PBMCs of CHB patients. Early changes in IFI16 mRNA, but not IFNB mRNA of PBMCs or serum IFI16, were correlated to HBeAg seroconversion of Peg-IFN-α2b therapy. An interaction between IFI16 and HBV DNA was detected in the PBMCs. In the cultured HepG2.2.15 and CEM-C7 cells, interferons resulted in the translocalization of IFI16 from the cytoplasm to the nucleus and inhibited IFI16 degradation. IFI16 of PBMCs may play a role in sensing HBV infection, and early change in IFI16 mRNA of PBMCs is valuable to predict HBeAg seroconversion in Peg-IFN-α2b treatment. The influences on IFI16 degradation and subcellular location may present a molecular mechanism of antiviral activity of interferon.

Restoration of HBV-specific CD8+ T-cell responses by sequential low-dose IL-2 treatment in non-responder patients after IFN-α therapy

Patients with chronic hepatitis B (CHB) undergoing interferon (IFN)-α-based therapies often exhibit a poor HBeAg serological response. Thus, there is an unmet need for new therapies aimed at CHB. This study comprised two clinical trials, including 130 CHB patients, who were treatment-naïve; in the first, 92 patients were systematically analyzed ex vivo for interleukin-2 receptor (IL-2R) expression and inhibitory molecules expression after receiving Peg-IFN-α-2b therapy. In our second clinical trial, 38 non-responder patients, in whom IFN-α therapy had failed, were treated with or without low-dose IL-2 for 24 weeks. We then examined the hepatitis B virus (HBV)-specific CD8+ T-cell response and the clinical outcome, in these patients. Although the majority of the participants undergoing Peg-IFN-α-2b therapy were non-responders, we observed a decrease in CD25 expression on their CD4+ T cells, suggesting that IFN-α therapy may provide a rationale for sequential IL-2 treatment without increasing regulatory T cells (Tregs). Following sequential therapy with IL-2, we demonstrated that the non-responders experienced a decrease in the numbers of Tregs and programmed cell death protein 1 (PD-1) expression. In addition, sequential IL-2 administration rescued effective immune function, involving signal transducer and activator of transcription 1 (STAT1) activation. Importantly, IL-2 therapy significantly increased the frequency and function of HBV-specific CD8+ T cells, which translated into improved clinical outcomes, including HBeAg seroconversion, among the non-responder CHB patients. Our findings suggest that sequential IL-2 therapy shows efficacy in rescuing immune function in non-responder patients with refractory CHB.

Effects of dyslipidemia on E antigen seroconversion of patients with chronic hepatitis B treated by nucleoside (acid) analogs

Background The prevalence of dyslipidemia in China is increasing annually. Current studies suggest that dyslipidemia affects the antiviral efficacy of hepatitis C virus (HCV) therapies, while recent studies suggest that serum lipids influence the response rates of chronic hepatitis B (CHB) patients receiving PEGylated interferon-alpha (Peg IFN-α) treatment. However, the role of dyslipidemia in the efficacy of nucleoside (acid) analogues (NAs) in CHB patients remains unclear. Methods From January 2010 to December 2013, data from 179 treatment-naive patients with CHB who were hepatitis B e antigen (HBeAg)-positive and had visited the first affiliated hospital of Wenzhou Medical University were assessed. Of these patients, 68 were assigned to the dyslipidemia group (diagnosed with CHB complicated with dyslipidemia) and 111 to the normolipidemic group. The following 3 treatment strategies were performed for all CHB patients over a 5-year period: lamivudine (LAM) plus adefovir dipivoxil (ADV) combination therapy, telbivudine (LdT) monotherapy, and entecavir (ETV) monotherapy. Serum assessments, blood biochemistry, HBV serological markers, HBV DNA before treatment and HBeAg serological conversion and virological responses at different timepoints after treatment were compared between the two groups. Measurement data were compared by τ tests and enumeration data by χ2 tests. Correlation analysis was performed using binary logistic regression analysis. Results The rates of HBeAg seroconversion in the dyslipidemia group at years 1, 2, 3, and 4 were 10.3, 13.2, 17.6, and 22.1%, respectively, which were not significantly lower than those of the normolipidemic group (11.7, 16.2, 18.0 and 33.3%; χ2 = 0.085, 0.293, 0.004, and 2.601, respectively; Ρ > 0.05). However, the rates of HBeAg seroconversion in the dyslipidemia group were significantly lower than those in the normolipidemic group at year 5 (27.9% vs. 43.2%, χ2 = 4.216, Ρ < 0.05). Univariate logistic regression analysis revealed significant differences in group, gender, PTA, ALT, AST, CR, and LDL-C between groups with and without seroconversion. Multivariate regression analysis demonstrated that dyslipidemia (OR = 1.993, Ρ = 0.038) and male gender (OR = 2.317, Ρ = 0.029) were risk factors associated with HBeAg seroconversion. Conclusions During antiviral therapy, dyslipidemia affects HBeAg seroconversion in CHB patients treated with NAs, but does not affect the virological response.

Association of Dyslipidemia With Nucleoside Analogue Treatment in HBeAg-Positive Chronic Hepatitis B Patients

BackgroundThe prevalence of dyslipidemia in China is increasing annually. Current studies suggest that dyslipidemia affects the antiviral efficacy of hepatitis C virus (HCV) therapies. Recent studies have shown that serum lipids influence the response rates of chronic hepatitis B patients receiving PEGylated interferon-alpha (Peg IFN-a) treatment. However, the role of dyslipidemia in the efficacy of nucleoside (acid) analogues in chronic hepatitis B patients has not been determined. Methods From January 2010 to December 2013, data from 179 treatment-naive patients with chronic hepatitis B (CHB) who were hepatitis B e antigen (HBeAg)-positive and visited the first affiliated hospital of Wenzhou Medical University were collected. Amongst them, 68 patients were diagnosed with CHB complicated with dyslipidemia (dyslipidemia group) whilst 111 patients comprised the lipid control group. Three treatment strategies were performed amongst the 179 CHB patients over a 5 year period. Treatments included combination therapy of lamivudine (LAM) plus adefovir dipivoxil (ADV), telbivudine (LdT) monotherapy or entecavir (ETV) monotherapy. Serum assessments, blood biochemistry, HBV serological markers, HBV DNA before treatment and HBeAg serological conversion and virological responses at different time points after treatment were compared between the two groups. Measurement data were compared using τ tests, whilst enumeration data were compared using c2 tests. Correlation analysis was performed using binary Logistic regression analysis. Results The rates of HBeAg seroconversion in the dyslipidemia group at years 1, 2, 3 and 4 were 10.3%, 13.2%, 17.6% and 22.1%, respectively, which were not significantly lower than those of the lipid control group 11.7%, 16.2%, 18.0% and 33.3%, (c2 = 0.085, 0.293, 0.004 and 2.601, respectively; R > 0.05). However, the rates of HBeAg seroconversion in the dyslipidemia group were significantly lower than those of the lipid control group at year 5 (27.9% vs 43.2%, c2 =4.216, R<0.05). Univariate logistic regression analysis showed significant differences in sex PTA, ALT, AST, CR and LDL-C. Multivariate regression analysis demonstrated that dyslipidemia (OR=1.993, R=0.038), and male gender (OR=2.317, R=0.029) were risk factors associated with HBeAg seroconversion.Conclusions During antiviral therapy, dyslipidemia affects HBeAg seroconversion in CHB patients treated with nucleoside (acid) analogues but does not affect the virological response.

Serum HBV RNA Predicts HBeAg Clearance and Seroconversion in Patients with Chronic Hepatitis B Treated with Nucleos(t)Ide Analogues

Background: To evaluate the predictive value of serum HBV DNA, HBV RNA, HBcrAg, HBsAg, intrahepatic HBV DNA, and cccDNA for HBeAg clearance and seroconversion during long-term treatment of nucleos(t)ide analogues (NAs) in patients with chronic hepatitis B (CHB).Method: A single center, prospective cohort of CHB patients enrolled between June 2007 and July 2008 was used for this study. Serum HBV RNA levels were retrospectively measured at baseline, 6, 12, 24, 36, 48, 60, 72, and 84 months post-NAs treatment. Serum HBsAg and HBcrAg levels were quantified at baseline, month 6, 60, and 72. Histological sample from liver biopsy at baseline and month 60 were analyzed for intrahepatic HBV DNA and cccDNA.Results: Eighty-three HBeAg patients were enrolled with an median follow-up time of 108 months (range 18-138 months). Of them, 53 (63.86%) patients achieved HBeAg clearance, and 37 (44.58%) achieved HBeAg seroconversion. Only baseline HBV RNA was independently associated with HBeAg clearance (OR=0.50, 95%CI 0.309-0.809, P=0.005) and seroconversion (OR=0.689, 95% CI 0.513-0.925, P=0.013). The independent negative association with HBeAg clearance and seroconversion remained for HBV RNA levels at month 6 (OR=0.42, 95%CI 0.248-0.714, P=0.001; OR=0.44, 95%CI 0.260-0.744, P=0.002) and month 12 (OR=0.39, 95%CI 0.253-0.592, P<0.001; OR=0.58, 95%CI 0.427-0.798, P=0.001). The AUC of baseline HBV RNA for predicting the HBeAg clearance and seroconversion were 0.81 (95%CI: 0.70-0.89) and 0.68 (95%CI: 0.56-0.78), respectively, higher than that of HBV DNA, HBsAg and HBcrAg.Conclusion: Lower serum HBV RNA at baseline, month 6 and 12 post NAs treatment could predict HBeAg clearance and seroconversion during long-term NAs treatment.

Early Serum HBV RNA Level in Combination With the HBeAg Response Can Effectively Predict the HBeAg Response in Patients on Nucleos(t)ide Analogue Therapy (ClinicalTrials.gov (NCT03909191)

Background: Serum HBV RNA level has the potential to monitor antiviral therapy in patients with chronic hepatitis B. This study aimed to explore serum HBV RNA dynamic change pattern and its predict value on the efficacy of 96 weeks nucleos(t)ide analogue (NA) treatment in HBeAg-positive and HBeAg-negative patients with chronic hepatitis B (CHB).Methods: A real-life cohort study of 78 patients with CHB on NA treatment was conducted. Dynamic change patterns of serum HBV RNA and correlation with other HBV markers in the early treatment period of 96 weeks of NA treatment in patients with CHB were determined and compared. The performance of serum HBV RNA on treatment efficacy was analyzed by receiver operating characteristic (ROC) analyses. Results: HBeAg-positive and HBeAg-negative patients with CHB had similar viral change patterns during NA treatment. Serum HBV RNA level was consistently correlated with HBeAg and HBsAg titers in HBeAg-positive patients during NA treatment, but serum HBV RNA was only moderately correlated with serum HBV DNA level in HbeAg-negative patients before treatment. Serum HBV RNA decreased more rapidly in patients with the HBeAg seroconversion (SC) response than in patients without the HBeAg SC response; it had good early discriminatory ability for the HBeAg response with area under the ROC curve (AUROC) of 0.70 and 0.730 at 12 and 24 weeks of treatment, respectively. The cutoff value of serum HBV RNA of 4.31 log cps/mL in combination with the HBeAg decrease degree of 1.55 at 24 weeks of treatment had a good two-way predictive capability for the HBeAg response (PPV%: 83.33% and NPV%: 81.25%), and the specificity was 96.30%. Conclusion: Serum HBV RNA level had early discriminatory ability for the HBeAg response. Early HBeAg response can improve the discriminatory ability of serum HBV RNA.