Antiviral prophylaxis for hepatitis B carriers improves the prognosis of diffuse large B‐cell lymphoma in Taiwan – a population‐based study

The prevention of hepatitis B virus (HBV) reactivation during rituximab treatment for diffuse large B‐cell lymphoma (DLBCL) is important in the HBV‐endemic area. This population‐based study examines the impact of antiviral prophylaxis for DLBCL patients with HBV infections. We identified 3702 adult patients with newly diagnosed DLBCL between 2011 and 2015 receiving R‐CHOP, R‐CVP, CHOP or CVP from the Taiwan Cancer Registry. We further stratified them into three groups: HBsAg‐negative patients (HBV‐negative, N = 2921), HBV carriers who received antiviral prophylaxis (HBV + Px, N = 711), and HBV carriers who did not receive antiviral prophylaxis (HBV + No Px, N = 70). HBV + Px patients were the youngest, and 69·4% received entecavir for antiviral prophylaxis. The median overall survival (mOS) of HBV‐negative and HBV + Px patients was similar (74·23 months and not reached, respectively). However, the mOS of HBV + No Px patients was only 35·61 months (P = 0·0028 compared with HBV + Px patients), indicating that antiviral prophylaxis improves OS in HBsAg‐positive DLBCL patients. The multivariate analysis showed that the HBV status and antiviral prophylaxis was an independent prognostic factor. In conclusion, our population‐based study illustrates the importance of antiviral prophylaxis in HBsAg‐positive DLBCL patients. Under antiviral prophylaxis, the survival of DLBCL patients with HBV infections was comparable to that of HBV‐negative patients.     

Protective efficacy and hepatitis B virus clearance in mice enhanced by cell‐mediated immunity with novel prime‐boost regimens

In this study, anti‐hepatitis B virus (HBV) immunity was evaluated in mice using several regimens of the HBV recombinant protein vaccine HBSS1 that expressed in CHO cells containing S (1‐223 aa) and preS1 (21‐47 aa) and recombinant adenovirus rAdSS1 vaccine. Further, the protective efficacy of these vaccine regimens was studied in a mouse model. High titres of antigen‐specific antibodies and neutralizing activity were elicited in mice after vaccination. However, robust multi‐antigen (preS1 and S)‐specific cell‐mediated immunity (CMI) was only detected in mice primed with HBSS1 and boosted with rAdSS1. Moreover, functional T‐cell responses with high levels of cytokines and antigen‐specific cytotoxic T‐cell responses (CD107a⁺CD8⁺) were also detected in the mice. Rapid clearance of hepatitis B surface antigen and HBV DNA in blood and significantly decreased hepatitis B envelope antigen levels were observed in mice immunized with the heterogeneous prime‐boost vaccine after hepatitis B virus challenge by hydrodynamic injection (HI) of pCS‐HBV1.3. The clearance of HBV correlated well with antigen‐specific CMI (Th1 and CTL responses) and cytokine profiles (IFN‐γ, TNF‐α, IL‐2) elicited by vaccination. Taken together, our results might contribute to the development of new human HBV vaccines and a better understanding of the mechanisms underlying immune protection and clearance of hepatitis B virus infection.     

Risk of hepatitis B virus reactivation in hepatitis B virus + hepatitis C virus‐co‐infected patients with compensated liver cirrhosis treated with ombitasvir,paritaprevir/r + dasabuvir + ribavirin

Hepatitis B virus may reactivate in patients with chronic hepatitis C treated with direct‐acting antivirals. The aim of this study was to investigate the risk of hepatitis B virus (HBV) reactivation in HBV + hepatitis C virus (HCV)‐co‐infected patients with compensated liver cirrhosis treated with paritaprevir/ombitasvir/ritonavir, dasabuvir with ribavirin. We reviewed prospectively gathered data from a national cohort of 2070 hepatitis C virus patients with compensated liver cirrhosis who received reimbursed paritaprevir/ombitasvir/r, dasabuvir with ribavirin for 12 weeks from the Romanian National Health Agency during 2015‐2016. Twenty‐five patients in this cohort were HBs antigen positive (1.2%); 15 untreated with nucleotide analogues agreed to enter the study. These patients were followed up: ALT monthly, serology for HBV and DNA viral load at baseline, EOT and SVR at 12 weeks. Hepatitis B virus (HBV)‐co‐infected patients were all genotype 1b and 52% females, with a median age of 60 years (51 ÷ 74); 76% were pretreated with peginterferon + ribavirin; 72% were with severe necroinflammatory activity on FibroMax assessment; 40% presented comorbidities; and all were HBe antigen negative. Hepatitis C virus (HCV) SVR response rate was 100%. Hepatitis B virus (HBV)‐DNA viral load was undetectable in 7/15 (47%) before therapy, and for the other 8 patients, it varied between below 20 and 867 IU/mL. Five patients (33%) presented virological reactivation (>2 log increase in HBV‐DNA levels) during therapy. One patient presented with hepatitis associated with HBV reactivation, and two started anti‐HBV therapy with entecavir. Hepatitis B virus (HBV) virological reactivation was present in 33% in our patients. Generally, HBV‐DNA elevations were mild (<20 000 IU/mL); however, we report one case of hepatitis associated with HBV reactivation.     

Low incidence of hepatitis B virus reactivation during chemotherapy among diffuse large B‐cell lymphoma patients who are HBsAg‐negative/ HBcAb‐positive: a multicenter retrospective study

Background: Reactivation of hepatitis B virus (HBV) is less common in lymphoma patients with prior resolved HBV infection [characterized by hepatitis B surface antigen (HBsAg)‐negative/hepatitis B core antibody (HBcAb)‐positive status] compared with chronic HBV infection (HBsAg positive) when receiving chemotherapy alone. The use of rituximab in chemotherapy regimen might increase the risk of HBV reactivation in patients with prior resolved HBV infection. However, the incidence of HBV reactivation is uncertain, and prophylactic antiviral treatment for this group of patients during rituximab‐containing chemotherapy is controversial. The objective of this study was to determine the incidence of HBV reactivation in HBsAg‐negative/HBcAb‐positive patients diagnosed of diffuse large B‐cell lymphoma (DLBCL) and treated with CHOP‐like or RCHOP‐like regimen. In addition, this study also aims to explore the relationship of HBV reactivation and HBV serology. Methods: Patients were identified using data from six university hospitals collected between January 1998 and November 2008. Four hundred and thirty‐seven patients with complete data were selected based on the diagnosis of CD20+ DLBCL, availability of HBV serum markers prior to initiation of chemotherapy and during the development of hepatitis, completion of at least four cycles of chemotherapy using CHOP‐like or RCHOP‐like regimen, and follow‐up for at least 6 months after completion of treatment. The characteristics of the HBsAg‐negative/HBcAb‐positive patients treated with CHOP‐like regimen were compared to those treated with RCHOP‐like regimen. Results: Eighty‐eight patients of the total 437 patients had pretreatment serology of prior resolved hepatitis B, with a prevalence of 20.1%. Among them, 45 patients received CHOP‐like regimen while 43 patients received RCHOP‐like regimen. Five patients developed hepatitis during treatment, two from CHOP group and three from RCHOP group. Only one patient treated with RCHOP had hepatitis associated with HBV reactivation, while the other four patients did not have evidence of HBV reactivation. Those four patients also demonstrated positive HBsAb at baseline, while the only patient who suffered from HBV reactivation had negative HBsAb status. This patient was successfully treated with antiviral medications. There were no statistically significant risk factors predictive of HBV reactivation. Conclusions: The present study revealed a low HBV reactivation rate of 2.3% in prior resolved hepatitis B among DLBCL patients undergoing RCHOP‐like therapy.     

HBV T1719G mutation reduced HBV replication through mutant Enh II and HBx protein in vitro

It was repeatedly reported that the hepatitis B virus (HBV) T1719G mutation was very common and related to progression and malignancy of liver disease. However, its effect on viral replication efficiency remains unclear. In this study, we aimed to evaluate the function and mechanisms of the T1719G mutation on viral replication capacity. Wild‐type and T1719G mutation‐bearing HBV1.2× plasmids were transfected into Huh7 and HepG2 cells, respectively, and HBV total RNA, 3.5 kb RNA and supernatant HBV DNA were assessed using real‐time PCR, hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels were measured by time‐resolved fluoroimmunoassay. In order to assess Enh II activity and the binding capacity of HNF3β to Enh II sequence, dual‐luciferase assay and Chromatin immunoprecipitation (ChIP)‐PCR were employed, respectively. Simultaneously, the HBx or HBx‐mut (T1719G) plasmid was co‐transfected to evaluate the effect of HBx on viral replication. Our results showed that the T1719G mutation impaired viral replication efficacy compared with the wild type both by reducing Enh II activity and binding capacity of HNF3β with Enh II. And such reduction caused by T1719G mutation could be rescued by HBx protein. Our results show that the T1719G mutation decreases HBV viral replication capacity possibly by mutant HBx protein and altered Enh II activity.     

Care quality and outcomes among US veterans with chronic hepatitis B in the hepatitis C direct‐acting antiviral era

Adherence to guideline‐recommended hepatitis B virus (HBV) care is suboptimal. We hypothesized that national hepatitis C eradication efforts during the era from 2015 to 2017 would improve the quality of care for cHBV given increased recognition and specialty referrals for liver disease. The study described herein is a retrospective cohort study of veterans with at least one positive HBsAg (HBsAg+) result from 1 January 2003 to 31 December 2017 using the VA Corporate Data Warehouse (CDW) analysed by era (2003‐2004, 2005‐2009, 2010‐2014, 2015‐2017). Relevant covariates such as HCV co‐infection, demographics, cirrhosis and baseline laboratory testing were obtained through previously validated approaches. We evaluated completion of process measures within 2 years of the index HBsAg + result: specialty care referral; testing of ALT, HBV‐DNA, HBeAg and anti‐HBe; testing for co‐infection and/or vaccination for HAV, HCV, HDV and HIV; and hepatocellular carcinoma (HCC) surveillance among those meeting criteria. We also measured use of antiviral therapy in appropriate candidates (ALT ≥ 2 × ULN, HBV‐DNA ≥ 2000 IU/mL). Of the 16 673 individuals with HBsAg + test results, 9,521 were confirmed as chronic HBV. Era‐related (Era 3:2010‐2014 vs Era 4:2015‐2017) increases in guideline‐recommended process measures included the following: outpatient visits with GI/ID specialists (78%‐89%), HBV‐DNA testing (73%‐79%), HDV testing (27%‐35%), appropriate HBV antiviral utilization (55%‐70%) and HCC surveillance (40%‐43%); all P < .0001. In the subset of HBV/HCV‐co‐infected patients, HCV DAA therapy was associated with a trend towards improved overall survival. In conclusion, the overall quality of care for HBV has significantly improved in the era of widespread HCV DAA therapy in an integrated health system possibly due to increased recognition and referral for liver disease.     

Hepatitis B virus and risk of non‐Hodgkin lymphoma: An updated meta‐analysis of 58 studies

Previous studies have focused on the relationship between hepatitis B virus (HBV) infection and non‐Hodgkin lymphoma (NHL). However, the results remain inconsistent and somehow conflicting in different subgroups. The aim of this study was to combine the findings of independent studies to comprehensively assess the association between HBV and NHL using a meta‐analysis. Relevant studies were identified through structured keyword searches in PubMed, EMBASE and the China National Knowledge Infrastructure (CNKI) database, and 58 studies with a total of 53 714 NHL cases and 1 778 591 controls were finally included. Pooled estimates indicated a significantly increased NHL risk in HBV‐infected individuals (summary odds ratio [sOR]: 2.50; 95% confidence interval [CI]: 2.20‐2.83) regardless of the study design (case–control studies: sOR: 2.47; 95% CI: 2.16‐2.82; cohort studies: sOR: 2.64; 95% CI: 1.78‐3.91). Considerable heterogeneity was observed across studies that was primarily attributed to the NHL subtypes (meta‐regression: P < .05). Overall, B‐cell NHL (sOR: 2.46; 95% CI: 1.97‐3.07) presented a stronger association with HBV infection than T‐cell NHL (sOR: 1.67; 95% CI: 1.34‐2.10). Within the B‐cell NHL subtypes, HBV infection was significantly associated with diffuse large B‐cell lymphoma (DLBCL, sOR: 2.06; 95% CI: 1.48‐2.88) and follicular lymphoma (FL, sOR: 1.54; 95% CI: 1.11‐2.12), but not with chronic lymphocytic leukaemia/small lymphocytic lymphoma (CLL/SLL) and Burkitt lymphoma. The results of this meta‐analysis support a positive link between HBV infection and NHL development. Further investigations for the mechanisms underlying HBV‐induced NHL are warranted.     

A novel in vivo model for studying conditional dual loss of BLIMP‐1 and p53 in B‐cells,leading to tumor transformation

The tumor suppressors B‐lymphocyte‐induced maturation protein‐1 (BLIMP‐1) and p53 play a crucial role in B‐cell lymphomas, and their inactivation contributes to the pathogenesis of a wide spectrum of lymphoid malignancies, including diffuse large B‐cell lymphomas (DLBCLs). Patients with activated B‐cell‐like (ABC) DLBCL may present with loss of BLIMP‐1, c‐Myc over‐expression, decreased p53, and poor prognosis. Nevertheless, there is a lack of in vivo models recapitulating the biology of high‐grade ABC DLBCL. We therefore aimed to develop an in vivo model aiming to recapitulate the phenotype observed in this cohort of patients. A Cre‐Lox approach was used to achieve inactivation of both p53 and BLIMP‐1 in murine B‐cells. Contextual ablation of BLIMP‐1 and p53 led to development of IgM‐positive B‐cell lymphoma with an aggressive phenotype, supported by c‐Myc up‐regulation, and accumulation of somatic mutations, as demonstrated by whole exome sequencing. Sensitivity of B‐tumor cells to BTK inhibition was demonstrated. This model mirrors what reported in patients with ABC DLBLC, and therefore represents a novel model for studying the biology of ABC‐DLBCL harboring the dual loss of BLIMP‐1/p53 and c‐Myc over‐expression.     

Hepatitis B virus‐specific T‐cell responses during IFN administration in a small cohort of chronic hepatitis B patients under nucleos(t)ide analogue treatment

The effect of pegylated interferon‐α (IFN) add‐on therapy on HBV‐specific T‐cell responses was evaluated in 12 patients with stable, undetectable hepatitis B virus (HBV) load under nucleos(t)ide analogue therapy. Peripheral blood mononuclear cells were isolated at week 0, 4, 8, 12, 24 and 48 of IFN add‐on therapy. Quantity and quality of circulating HBV S‐ and core‐specific CD4 and CD8 T cells were analysed ex vivo by flow cytometry. HBV S‐ and core‐specific CD4 T‐cell numbers modestly increased within 8 weeks of IFN administration (P = 0.0391 and P = 0.0195), whereas HBV‐specific CD8 T cells in general showed only minor changes under IFN add‐on therapy. Functionality of HBV‐specific CD4 but not CD8 T cells positively correlated with serum transaminase activity. In addition, we observed an increase in CD4 T cells producing tumour necrosis factor‐α (TNFα) without antigen restimulation (P = 0.0039), which correlated with elevated transaminases. During IFN add‐on therapy, two patients developed an anti‐HBs seroconversion, only one of whom showed a relevant increase in HBV‐specific T cells. In conclusion, IFN add‐on therapy of chronic hepatitis B increased HBV‐specific T‐cell responses and affected a previously unrecognized TNFα‐monofunctional CD4 T‐cell population. Although the observed T‐cell responses did not correlate with HBsAg seroconversion, we expect additional insights into the immunopathogenesis of hepatitis B, following the characterization of the newly identified TNF α‐monofunctional T‐cell population.     

乙型肝炎病毒X基因克隆及表达的研究

目的克隆、表达并纯化乙型肝炎病毒Ｘ抗原,应用 ｗｅｓｔｅｒｎ印迹分析该抗原与乙型肝炎患者体内抗Ｘ抗体的特异性反应。方法应用基因扩增技术分离此蛋白的结构基因,采用基因工程的方法,应用融和表达载体ｐＧＥＸ－２Ｔ,重组、克隆得到带有ＨＢＸ特异抗原基因的重组菌株,并应用ｗｅｓｔｅｒｎ印迹的方法检测乙型肝炎患者体内的抗Ｘ抗体。结果ｗｅｓｔｅｒｎ印迹分析表明,该表达产物可与ＨＢＶ感染患者体内的抗Ｘ抗体特异性结合。结论纯化后的ＨＢＸ抗原可作为检测乙型肝炎患者抗Ｘ抗体的特异性抗原。     

HBV‐specific T‐cell responses in healthy seronegative sexual partners of patients with chronic HBV infection

Summary. The hepatitis B virus (HBV) is frequently transmitted by sexual intercourse. Thus, HBV‐guidelines recommend vaccination. However, we have identified healthy hepatitis B surface antigen and anti‐HBc‐negative unvaccinated sexual partners of patients with chronic hepatitis B. We investigated whether HBV‐specific cellular immune responses were present that could explain the apparent protection against HBV infection. In six anti‐HBc‐negative HBV‐exposed sexual partners, HBV‐specific T‐cell responses were studied by proliferation assay and cytometric bead array after stimulation with 74 overlapping peptides spanning the HBV core, pre‐S and S‐encoding regions. Eleven HBV‐unexposed individuals served as negative controls. HBV‐DNA was undetectable in serum and peripheral blood mononuclear cells in all cases. HBV‐specific cytokine secretion was observed in 4/6 seronegative partners, but only in 1/11 controls. Proliferative responses were detectable in 5/6 partners and 0/11 controls. HBV‐specific cytokine secretion exists in healthy seronegative virus‐exposed individuals. HBV core‐directed immune responses indicate past, but controlled viral replication. T‐cell immunity may prevent clinical manifestation of HBV infection in the absence of humoral immunity.     

CD30 expression in de novo diffuse large B‐cell lymphoma: a population‐based study from British Columbia

Diffuse large B‐cell lymphoma (DLBCL) is a heterogeneous disease with variable therapeutic responses and alternative therapies are needed for patients with unfavourable treatment outcomes after standard treatment with R‐CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone). One promising candidate is brentuximab vedotin, an antibody‐drug conjugate targeting CD30‐expressing cells. However, CD30 (TNFRSF8) expression patterns in DLBCL are not well described thus far. Here, we examined CD30 expression in a population‐based cohort of immunocompetent patients from British Columbia with de novo DLBCL using immunohistochemistry. 385 cases of formalin‐fixed paraffin‐embedded DLBCL in tissue microarrays were evaluated. 95 cases (25%) harboured CD30+ tumour cells. Using a > 0% cut‐off, CD30 expression was predictive of superior 5‐year progression‐free survival within R‐CHOP treated germinal centre B‐cell‐like (GCB) DLBCL (86% vs. 64%, P = 0·020), which was independent of the International Prognostic Index. Epstein‐Barr virus (EBV) was identified in 11 (3%) cases, all of which were non‐GCB (P = 0·001) and almost exclusively positive for CD30 expression (10/11) (P < 0·001). We conclude CD30 is expressed in a substantial proportion of DLBCL and CD30 immunohistochemistry may be a useful prognostic marker in R‐CHOP treated GCB‐DLBCL. The significant association of CD30 with EBV‐positive non‐GCB DLBCL suggests a distinct pathobiology for these cases.     

乙型肝炎患者血清Pre-S-2抗原的意义

目的研究ＰｒｅＳ２抗原与乙型肝炎患者ＨＢＶ标记的关系．方法血清ＨＢｓＡｇ（＋），ＨＢｅＡｇ（＋），ＨＢｃＡｂ（＋）的乙型肝炎患者２６例，血清ＨＢｓＡｇ（＋），ＨＢｅＡｂ（＋），ＨＢｃＡｂ（＋）的乙型肝炎患者４７例及健康献血者２０例，血清用ＲＩＡ法检测ＰｒｅＳ２抗原及用ＰＣＲ法检测ＨＢＶＤＮＡ．结果血清ＨＢｓＡｇ（＋），ＨＢｅＡｇ（＋），ＨＢｃＡｂ（＋）的乙型肝炎患者２６例，ＰｒｅＳ２抗原与ＨＢＶＤＮＡ均阳性（１００％）；血清ＨＢｓＡｇ（＋），ＨＢｅＡｂ（＋），ＨＢｃＡｂ（＋）的乙型肝炎患者４７例，ＰｒｅＳ２抗原３０例阳性（６３８％），１７例阴性（３６２％），ＨＢＶＤＮＡ３２例阳性（６８１％），１５例阴性（３１９％），ＰｒｅＳ２抗原与ＨＢＶＤＮＡ均阳性２８例（５９６％），均阴性１４例（３００％）．健康献血者２０例，ＰｒｅＳ２抗原阳性１例（５０％），阴性１９例（９５０％），ＨＢＶＤＮＡ阳性２例（１００％），阴性１８例（８００％），ＰｒｅＳ２抗原及ＨＢＶＤＮＡ均阳性０例（０％），均阴性１８例（８００％）．结论ＰｒｅＳ２抗原可作为预测慢性乙型肝炎患者病情活动与传染的标志．     

Hepatitis B virus‐related hepatocellular carcinoma in the era of antiviral therapy: The emerging role of non‐viral risk factors

Hepatocellular carcinoma (HCC), one of the major malignant lethal tumours, is most prevalent in Asian patients with chronic hepatitis B virus (HBV) infection. Both viral and non‐viral factors contribute to the development of HCC. It is established that viral factors associated with HBV DNA level, HBV genotype, designated gene mutation, HBV DNA integration, HBx protein, hepatitis B surface antigen (HBsAg), hepatitis B core‐related antigen (HBcrAg) and HBV RNA are correlated with hepatocarcinogenesis. Before the introduction of antiviral therapy, viral factors once attracted more attention during the development of HCC. With the widespread use of antiviral therapy, predominantly nucleos(t)ide analogues (NAs), most patients with chronic hepatitis B (CHB) have achieved sustained viral control. The role of non‐viral factors, especially modifiable factors, is anticipated to be reinforced in the future. Herein, we reviewed the modifiable non‐viral risk factors of HBV‐related HCC, in the hope of providing substantial evidence for further development of novel precautionary measures for HCC. In addition, the therapeutic interventions for reducing the risk of HCC, like potential conventional pharmaceutical interventions and lifestyle modification are also discussed in this review. Future studies that would explore the specific mechanism of HBV‐related HCC development in patients with satisfactory viral control and related precision treatment are warranted.     

Efficacy and safety of telbivudine and tenofovir disoproxil fumarate in preventing hepatitis B vertical transmission: A real‐life practice

Mother‐to‐child transmission (MTCT) is a major obstacle in the elimination of hepatitis B virus (HBV) infection. Telbivudine (LdT) and tenofovir disoproxil fumarate (TDF) are the two most common antiviral medicines for preventing MTCT. However, the efficacy and safety of LdT and TDF in preventing HBV vertical transmission during the second to third trimester have not been compared rigorously. Therefore, we carried out a prospective multicentre cohort study of chronic hepatitis B in mothers with HBV DNA > 10⁶ IU/mL, receiving LdT or TDF during the second to third trimester. Among the 893 mothers enrolled, 857 (LdT/TDF/untreated group (NTx) = 396/325/136) completed consecutive follow‐up with 854 infants (LdT/TDF/NTx = 395/323/136). LdT and TDF treatment resulted in a similar decrease of HBV DNA in mothers at delivery. Multivariate analysis indicated that only HBsAg titre at the baseline correlated with viral DNA decrease (P = 0.015). With intention‐to‐treat analysis, MTCT rates in the LdT, TDF and NTx group were 4.41%, 2.42% and 22.08%, respectively. An increasing vertical transmission rate was found to be closely associated with higher HBsAg titre, 5.32% and 17.65% infection rate was estimated in infants born to mothers with HBsAg > 4 and >5 log₁₀ IU/mL, respectively. No serious side effects were reported in either mothers or infants. LdT and TDF treatments were well tolerated and showed comparable efficacy in reducing MTCT. Higher risk of MTCT was shown in pregnant women with HBsAg > 4 log₁₀IU/mL.     

Hepatitis B Virus Induces Microtubule Stabilization to Promote Productive Infection through Upregulating Microtubule-associated Protein 1S

Background and AimsContinuous release and transmission of hepatitis B virus (HBV) is one of the main factors leading to chronic hepatitis B (CHB) infection. However, the mechanism of HBV-host interaction for optimal viral transport is unclear. Hence, we aimed to explore how HBV manipulates microtubule-associated protein 1S (MAP1S) and microtubule (MT) to facilitate its transport and release.MethodsThe expression of MAP1S or acetylated MT was investigated by immunofluorescence, RT-PCR, immunoblotting, and plasmid transfection. MAP1S overexpression or knockdown was performed by lentiviral infection or sh-RNA transfection, respectively. HBV DNA was quantified using q-PCR.ResultsSignificantly higher level of MAP1S in HepG2215 cells compared with HepG2 cells was detected using RT-PCR (p<0.01) and immunoblotting (p<0.001). Notably, stronger MAP1S expression was observed in the liver tissues of patients with CHB than in healthy controls. MAP1S overexpression or knockdown demonstrated that MAP1S promoted MT acetylation and reduced the ratio of HBV DNA copies inside to outside cells. Further, transfection with the hepatitis B virus X protein (HBx)-expressing plasmids induced significantly higher level of MAP1S than that in controls (p<0.0001), whereas HBVX⁻ mutant-encoding HBV proteins (surface antigen, core protein, and viral DNA polymerase) hardly affected its expression.ConclusionsThese results demonstrate that HBx induces the formation of stable MTs to promote the release of HBV particles through upregulating MAP1S. Thus, our studies delineate a unique molecular pathway through which HBV manipulates the cytoskeleton to facilitate its own transportation, and indicate the possibility of targeting MAP1S pathway for treatment of patients with CHB.     

Pre-S2抗原在乙肝病毒隐性感染中的地位与应用研究

探讨Pre-S2抗原在乙型肝炎病毒的各类隐性感染中的地位及科研、临床应用价值。以乙肝病毒隐性感染者作为研究对象,采用ELISA法,同时检测感染者的Pre-S2抗原和“乙肝血清标志物(HBV M)”,然后进行各类感染者之间的比较、分析。在乙肝病毒隐性感染者人群中,HBeAg阳性者、HBeAb阳性者的Pre-S2抗原阳性率分别为93．10％、60．00％;“乙肝大三阳”、“小三阳”者的Pre-S2抗原阳性率分别为92．31％、62．50％。Pre-S2抗原在判断乙肝病毒复制、隐性感染者血清的传染性强度方面,有很高使用价值和补缺作用;在判断隐性感染者的发展转化方面,亦有重大参考意义。     

Reactivation of hepatitis B virus during interferon‐free therapy with daclatasvir and asunaprevir in patient with hepatitis B virus/hepatitis C virus co‐infection

Direct‐acting antiviral agents (DAA) for hepatitis C virus (HCV) are not effective for hepatitis B virus (HBV), which may be suggestive of reactivation of anti‐HBe hepatitis during interferon (IFN)‐free DAA therapy in HBV/HCV co‐infected patients with inactive HBV. A 69‐year‐old male patient was diagnosed with chronic hepatitis due to HBV/HCV co‐infection with serum levels of alanine aminotransferase (ALT) of 94 U/L, HCV RNA of 4.2 log IU/mL and HBV DNA of 2.5 log copies/mL. HCV was thought to be responsible for the hepatitis activity because of low level of HBV core‐related antigen (3.1 log U/mL). He was treated with combination therapy of daclatasvir and asunaprevir. Serum ALT gradually increased, and reached 237 U/L on day 43 in spite of undetectable HCV RNA. Serum HBV DNA was increasing to 7.0 log copies/mL at that time. The treatment was stopped due to suspicion of drug‐induced liver injury and/or HBV reactivation. Administration of entecavir reduced HBV DNA levels, followed by improvement in ALT levels. This report proposes that close monitoring of HBV DNA during the anti‐HCV DAA therapy and the commencement of anti‐HBV therapy with nucleoside analogs after the increase of HBV DNA should be considered in patients with HBV/HCV co‐infection.