Research advances in persistent hepatitis B virus infection and its mechanism / 临床肝胆病杂志

Hepatitis B virus (HBV) belongs to the Hepadnaviridae family and can cause acute and chronic hepatitis, liver cirrhosis, and even liver cancer in humans. Current antiviral drugs cannot completely eliminate HBV in liver cells and thus it is difficult to achieve a curative effect. In recent years, the mechanism of persistent HBV infection has attracted wide attention, which mainly involves host and virus. This article elaborates on the research advances in persistent HBV infection from the aspect of virus, including covalently closed circular DNA, HBV particles, and HBV components.

The prevalence of occult hepatitis B infection among the blood donors in a tertiary care hospital, Puducherry

Occult hepatitis B infection (OBI) is a cause of concern while screening the blood donors to prevent transfusion-related transmission of infection. This study was conducted to assess the prevalence of OBI using total anti-HBc by ELISA and DNA detection by real time polymerase chain reaction (PCR). The samples included were negative for HBs Ag by ELISA. Out of 1102 samples tested, 156 were positive for total anti-hepatitis B core antigen and 52/156 by real-time PCR. Overall, the prevalence was found to be 4.71% (52/1102). The results indicate that nucleic acid-based testing should be an essential part of screening procedure to prevent missing of OBI.

Anti-HBc levels in patients with different natural history of chronic HBV infection and cirrhosis and its clinical significance / 重庆医学

Objective To investigate the serum anti-HBc level in patients with different natural history of chronic hepatitis B virus (HBV) infection and cirrhosis,and its clinical value for distinguishing the natural history statue.Methods A total of 160 patients with chronic HBV infection from March 2015 to June 2016 were enrolled,and they were divided into immune tolerance group (n=43),HBeAg-positive chronic hepatitis B (CHB) group (n=37),inactive carrier group (n=39) and HBeAg-negative CHB group (n=41).A total of 44 patients with HBeAg-positive cirrhosis and 46 patients with HBeAg-negative cirrhosis were enrolled too.The general conditions data were collected,and HBsAg,HBeAg,anti-HBc,HBV DNA load and HBV genotype were detected.The associations between anti-HBc level and clinical parameters were analyzed,and the diagnostic value of anti-HBc for distinguishing different natural histories was analyzed.Results The anti-HBc levels in different natural history from high to low were as following: HBeAg-positive CHB group (4.22±0.68)log10 IU/mL,HBeAg-negative CHB group (3.89±0.88)log10 IU/mL,inactive carrier group (3.07±0.68)log10 IU/mL and immune tolerance group (2.88±0.82)log10 IU/mL.The anti-HBc levels in HBeAg-positive and HBeAg-negative cirrhosis patients were (3.04±0.82) and (3.15±0.86) log10 IU/mL,respectively.In HBeAg-positive CHB group,the anti-HBc was positively associated with ALT (r=0.353,P=0.032) and AST (r=0.421,P=0.009).In HBeAg-negative CHB group,the anti-HBc was positively associated with HBV DNA (r=0.343,P=0.028),ALT (r=0.458,P=0.003) and AST (r=0.495,P=0.001).The AUC of anti-HBc used to distinguish immune tolerance from HBeAg-positive CHB was 0.903,and the AUC used to distinguish inactive carrier from HBeAg-negative CHB was 0.833.Conclusion Anti-HBc levels in different natural history of chronic HBV infection are significantly different,and anti-HBc could be used to distinguish the natural history statue of chronic HBV infection with a higher diagnostic value than HBsAg.

Hepatitis B core antigen promotes invasion of hepatocellular carcinoma cell line HepG2.2.15 via Toll-like receptor 4 / 中华肝脏病杂志

Objective@#To investigate the effect of hepatitis B core antigen (HBcAg) in promoting the invasion of hepatitis B virus (HBV)-related hepatocellular carcinoma cell line HepG2.2.15 and the role of Toll-like receptor 4 (TLR4) in the mechanism.@*Methods@#TLR4 mRNA and protein expression in HepG2 cells and HepG2.2.15 cells was measured by reverse transcription real-time PCR and Western blot analysis, respectively. HepG2.2.15 cells were transfected with TLR4 specific small interfering RNA (siRNA) to silence TLR4 expression, and stimulated by recombinant HBcAg in culture. The invasion of cells was measured by Transwell invasion assay. The expression of TLR4 signaling pathway-related proteins in the cultured cells and proinflammatory cytokines in the supernatant was also determined. The student’s t-test, one-way ANOVA, and SNK-q test were used for statistical analysis.@*Results@#TLR4 mRNA and protein expression in HepG2.2.15 cells was significantly higher than that in HepG2 cells. TLR4 siRNA transfection remarkably down-regulated TLR4 expression in HepG2.2.15 cells. Inhibiting TLR4 expression and/or HBcAg stimulation did not affect the proliferation of HepG2.2.15 cells. However, HBcAg stimulation significantly increased the invasion ability of HepG2.2.15 cells and the secretion of proinflammatory cytokines [including interferon (IFN)-γ, interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α]. Inhibiting TLR4 expression significantly reduced HBcAg-induced cellular invasion. Meanwhile, HBcAg stimulation elevated the expression of MyD88 and TRIF in HepG2.2.15 cells. TLR4 silencing inhibited HBcAg-induced increase in the expression of MyD88, while it showed no significant impact on TRIF expression.@*Conclusion@#HBcAg can promote the invasion of HepG2.2.15 cells. The TLR4/MyD88 signaling pathway may be involved in this process by inducing the expression of proinflammatory cytokines.

Expression intensity and clinical significance of intrahepatic hepatitis B surface antigen and hepatitis B core antigen in 994 patients with chronic hepatitis B virus infection / 中华临床感染病杂志

Objective To investigate the intensity of HBsAg and HBcAg expression in liver tissue of patients with chronic hepatitis B virus (HBV) infection and its clinical significance.Methods A total of 994 HBV infected patients underwent liver biopsy and histopathological examination.The expression of HBsAg and HBcAg in liver tissue was detected by histoimmunochemistry.Patients were divided into HBeAg (+)/HBVDNA(+), HBeAg(-)/HBV DNA(+) and HBeAg(-)/HBV DNA(-) groups according to HBeAg and HBV DNA levels;patients were divided into ＜2 × normal (ULN) group, 2-＜5 × ULN groupand ≥5 × ULN group according to the alanine aminotransferase (ALT) levels.The histologic activity (A), fibrosis (F), the expression of HBsAg and HBcAg in liver tissue and their correlations with clinical features were analyzed.Logistic regression analysis was used to study the factors affecting the expression of HBsAg and HBcAg in liver tissue.Results Among 994 HBV infected patients, 941 cases (94.67％) were intrahepatic HBsAg positive and 553 cases (55.63％) were intrahepatic HBcAg positive;403 cases (40.85％) were ≥A2 in histologic activity and 371 cases (36.09％) were ≥F2 in fibrosis.The degree of A and F was the highest in HBeAg (-) / HBV DNA (+) group, followed by HBeAg (-) / HBV DNA (-) group, and was the lowest in HBeAg (+) / HBV DNA (+) group.The intensity of intrahepatic HBsAg expression was significantly different among three groups (x2 =6.299, r =-0.760, P ＜ 0.05), however, the difference was not showed in pairwise comparisons.The difference of intrahepatic HBcAg intensity among three groups was statistically significant (x2 =282.995, r =-0.645, P ＜ 0.01), the intensity was the highest in HBeAg (+) / HBV DNA (+) group and the lowest in HBeAg (-) / HBV DNA (-) group.The constituent ratio of HBeAg positive and HBV DNA level were higher and the average age was lower in intrahepatic HBsAg positive group than those in HBsAg negative group.The constituent ratio of positive HBeAg, the levels of ALT, AST, PLT and HBV DNA were higher and the average age, the average FIB-4 level were lower in intrahepatic HBcAg positive group than those in HBcAg negative group.The HBV DNA level was an independent risk factor for intrahepatic HBsAg intensity, and the HBeAg positive and HBV DNA level were independent risk factors for intrahepatic HBcAg intensity.There were no significant differences in A and F among different groups of intrahepatic HBsAg intensity (x2 =1.943 and 2.630, both P ＞ 0.05).There was significant difference in F among different groups of intrahepatic HBcAg intensity (x2 =12.352, P ＜ 0.01), but not in A.The degree of F was the highest in intrahepatic HBcAg negative group.There was significant difference in intrahepatic HBcAg intensity among different groups of ALT level (x2 =16.349, P ＜ 0.01), but not in intrahepatic HBsAg intensity.The intrahepatic HBcAg intensity in ALT ＜ 2 × ULN group was lower than that in other two groups.Conclusions Most of patients with chronic HBV infection are intrahepatic HBsAg positive and more than half of them are intrahepatic HBcAg positive.The intrahepatic HBsAg intensity is not associated with A and F, but correlates with HBV DNA level.The intrahepatic HBcAg intensity is not associated with A, but it is negatively correlated with F and positively correlated with positive HBeAg expression, HBV DNA level and ALT level.

Clinical study of de novo hepatitis B virus infection after pediatric living liver transplantation / 器官移植

Objective To investigate the clinical characteristics,prevention and treatment strategy of de novo hepatitis B virus (HBV)infection after pediatric living liver transplantation.Methods In total,106 pediatric recipients undergoing living liver transplantation in Organ Transplantation Center of Affiliated Beijing Friendship Hospital of Capital Medical University and Organ Transplantation Center of Tianjin First Center Hospital from July 2010 to July 2014 were enrolled in this study.All surgeries were performed by the same surgical team.According to preoperative test outcomes of donor HBV serological markers,all recipients were divided into the positive (n =45)and negative (n =61)antibody to hepatitis B core antigen (anti-HBc)donor liver groups (positive group and negative group),and the prevalence of de novo HBV infection was compared between two groups.The risk factors of de novo HBV infection in the positive group were analyzed to elucidate the clinical characteristics of de novo HBV infection in affected children.Results The incidences of de novo HBV infection in positive and negative group were 18% (8 /45 )and 2% (1 /61 )respectively.The risk factors of de novo HBV infection in recipients with positive anti-HBc were negative anti-HBs before transplantation and absence of antiviral therapy post-transplantation in recipients (both in P <0.05 ).The median interval between time of onset and time of liver transplantation was 12 months (8-48 months).Seven cases were treated with lamivudine and the remaining two cases were left untreated.All nine recipients survived.Conclusions Application of positive anti-HBc donor liver have a risk of HBV infection in recipients after pediatric liver transplantation.Absence of postoperative nucleoside analogue therapy and negative anti-HBs before transplantation acts as risk factors of de novo HBV infection in the recipients with positive anti-HBc donor liver.After liver transplantation,nucleoside analogue therapy is recommended for the pediatric recipients with positive anti-HBc donor liver to prevent the incidence of de novo HBV infection.Besides,hepatitis B vaccine should be administered prior to liver transplantation.

Hepatitis B Core Antigen Expression in Hepatocytes Reflects Viral Response to Entecavir in Chronic Hepatitis B Patients

BACKGROUND/AIMS: Hepatitis B core antigen is known to be a major target for virus-specific T cells and also reflects the progression of liver dissease and viral replication. Hepatitis B core antigen expression in hepatocytes leads to altered histological activity, viral replication, and immune response. The purpose of this study is to evaluate whether the topographical distribution of hepatitis B core antigen expression can predict the viral response to entecavir in patients with chronic hepatitis B. METHODS: We enrolled 91 patients with treatment-naive chronic hepatitis B. All the patients underwent liver biopsy, and the existence and pattern of hepatitis B core antigen evaluated by immunohistochemistry. All patients received 0.5 mg of entecavir daily following a liver biopsy. We checked the viral response at 3, 6, and 12 months during antiviral therapy. RESULTS: Of the 91 patients, 64 (70.3%) had hepatitis B core antigen expression. Of the subcellular patterns, the mixed type was dominant (n=48, 75%). The viral response was significantly higher in the hepatitis B core antigen-negative group than in the hepatitis B core antigen-positive group (88.9% and 54.7%, respectively; p=0.001) after 12 months of entecavir therapy. CONCLUSIONS: Chronic hepatitis B patients who are hepatitis B core antigen-negative have a better response to entecavir therapy than do hepatitis B core antigen-positive patients.

Reactivation of Hepatitis B Virus Following Systemic Chemotherapy for Malignant Lymphoma / 대한내과학회지

BACKGROUND/AIMS: Reactivation of hepatitis B virus (HBV) has been reported in HBV surface antigen (HBsAg)-positive patients undergoing chemotherapy, as well as HBsAg-negative patients with antibodies against HBV core antigen (HBcAg) and/or HBsAg (HBsAb). Chemotherapy-including rituximab-has recently been identified as a predictive factor for HBV reactivation in HBsAg-negative patients with malignant lymphoma. The aim of our study was to identify the factors predictive of HBV reactivation after chemotherapy in patients with malignant lymphoma. METHODS: We conducted a retrospective analysis of medical records from patients diagnosed with malignant lymphoma at Gachon University Gil Medical Center in City, County from January 2005 to December 2010. We subsequently determined HBsAg, HBsAb and anti-HBc status in the 196 patients treated with chemotherapy. RESULTS: The mean age of the patients was 57.3 +/- 14.5 years; 56.3% were male. A total of 172 of 196 (88%) patients in the study population were HBsAg (+) prior to chemotherapy. Three patients (3/11, 27.3%) in the HBsAg (+) group had confirmed HBV reactivation after chemotherapy. In addition, 26 of 196 (13%) patients in the study population tested HBcAg (+) positive prior to chemotherapy. One patient (1/15, 6.7%) in the HBsAg (-)/HBcAb (+) group had confirmed HBV reactivation. In the four patients with HBV reactivation, infection was resolved after treatment with 0.5 mg entecavir or 100 mg lamivudine. CONCLUSIONS: Reactivation of HBV after systemic chemotherapy can occur in HBsAg (-) patients. We recommend that malignant lymphoma patients undergoing chemotherapy be screened for HBV infection status, including HBcAg, and followed closely to prevent HBV reactivation.

Relation of hepatitis B core antigen expression with histological activity, serum HBeAg, and HBV DNA levels.

Background: Hepatitis B virus (HBV) is a potentially life-threatening liver infection which may progress to liver failure and cirrhosis. Intrahepatic expression patterns of viral antigens detected by immunohistochemistry may have prognostic implications in disease process. Aim: In this study, we aimed to investigate the relationship between the HBV core antigen (HBcAg) expression and histological activity index (HAI), fibrosis, serum hepatitis B e-antigen (HBeAg) status and HBV DNA levels in patients with chronic HBV infection. Materials and Methods: A total of 114 liver biopsies from patients with chronic HBV infection were included in the study. Immunohistochemical expression of HBcAg and its relation with HAI, fibrosis, serum alanine aminotransferase (ALT) levels, HBeAg status and HBV DNA levels were assessed. Results: The presence of nuclear expression of HBcAg did not show any correlations with ALT levels, HAI and fibrosis score. When the groups were categorized according to the HBeAg status, nuclear HBcAg expression was found to be high in HBeAg positive patients. However, HBcAg nuclear expression showed significant correlations with HBV DNA levels and fibrosis scores in HBeAg negative but not HBeAg positive patients. HBV DNA levels were also significantly associated with HAI and fibrosis scores in HBeAg negative patients. Conclusions: Significant differences found between HBeAg positive and negative patients suggest that HBeAg negative disease is different from HBeAg positive disease, and also point outs that in HBeAg negative disease, patients with nuclear HBcAg expression and increased levels of HBV DNA levels are at a higher risk of developing progressive liver disease.

Clinical and pathological characteristics of patients with chronic hepatitis B virus infection in immune tolerant phase / 中华传染病杂志

Objective To investigate the clinical and pathological characteristics of patients with chronic hepatitis B virus(HBV)infection in immune tolerant phase.Methods Ninety-eight chronic HBV carriers in immune tolerant phase were enrolled in this study.The age,gender,serum HBV DNA level,hepatic inflammatory activity and fibrosis,hepatic HBsAg and HBcAg expressions were analyzed.The grade of inflammatory activity and stage of fibrosis were also compared in patients with different levels of serum alanine aminotransferase(ALT).Data analysis was done by chi-square test. Results In 98 patients,83(84.7%)were＜30 years old and 15(15.3%)were≥30.Patients whose mother was HBsAg positive were 48.0%.High levels of serum HBV DNA were found in all patients, with 78.5% were＞1×107 copy/mL.Only 5 cases(5.1%)were G0 of the inflammatory grade;whereas,64 cases(65.3%)were G1,29(29.6%)were G2.There were 56 patients(57.1%)had no significant liver fibrosis;and 23 cases(23.5%)were S1,14(14.3%)were S2,5(5.1%)were S3;none of patients had cirrhosis.The HBsAg and HBcAg in liver tissues were positive in 79(80.6%)and 80(81.6%)cases,respectively.The fibrosis stages of patients with higher ALT levels were significantly greater than patients with lower ALT levels(X2=8.112 3,P=0.043 7).Conclusions Most of patients with chronic HBV infection in immune tolerant phase present mild inflammation in liver,some of them have already developed fibrosis.Therefore,liver pathology is recommended for these patients to help understand the patients' conditions and make correct therapeutic decisions.

The Degrees of Hepatocyte Cytoplasmic Expression of Hepatitis B Core Antigen correlate with Histologic Activity of Liver Disease in the Young Patients with Chronic Hepatitis B Infection

Subcellular localizaton of HBcAg have been found to be related to the activity of liver disease and HBV replication. The aim of this study was to determine whether the degree of expression of HBcAg in the hepatocyte nucleus and cytoplasm reflects the level of viral replication and histological activity in chronic HBV infection. A total of 102 patients with biopsy proven chronic hepatitis B were included. There was a highly significant correlation between the levels of HBV DNA in serum and the degree of expression of HBcAg in the nucleus for HBeAg-positive(p=0.000) and negative patients(p=0.04). There was a highly significant, correlation between the degrees of expression of HBcAg in hepatocyte cytoplasm and histologic activities (p<0.01) for HBeAg-positive patients. The degrees of expression of HBcAg in the hepatocyte cytoplasm correlated positively with the lobular activities (p<0.01), but not correlated with the portal activity and fibrosis for HBeAg-negative patients. In conclusion, in the young patients with chronic B viral hepatitis, the degree of expression of HBcAg in the hepatocyte nucleus may affect viral load, and the degree of expression of HBcAg in the hepatocyte cytoplasm may affect histologic activities of liver disease.

Development of hepatitis virus C therapeutic vaccine with hepatitis virus B core antigen us the carrier / 中华传染病杂志

Objective To invesgate hepatitis B c antigen(HBeAg)as the carrier to construct mixed hepatitis C virus(HCV)therapeutic vaccine.Methods Fused the pTrc-core gene with two synthetic T-epitope antigen gene of HCV,expressed the plasmids pTrc-core-T1 and pTrc-core-T2, applied sucrose density gradient centrifugation to get the fusion protein HBcAg-T1 and HBcAg-T2, dialysised and concentrated the protein,mixed and immunized them in mice using the protein HBcAg (expressed by pTrc-core)as control.The tumor regression trial in mice was evaluated at appropriate time.After immunized four times,got the blood and spleen of mice.Interleukin(IL)-12 in serum were detected by enzyme-linked immunosorbent assay(ELISA),nonspecific T lymphocytes prolifera- tion response of splenic lymphocytes was respectively examined by thiazolyl blue(MTT)assay.T cell subset of splenic lymphoeytes,IL-5 in serum,IL-4 and interferon(IFN)-?in lymphocyte were evalu- ated by FACS.Results Tumor regression trial showed the experimental group formed only one tumor(diameter=0.1 cm),smaller than the T_1T_2 peptides group(diameter=0.9 cm)and blank group(diameter=1.3 cm).FACS indicated that CD8~+ T cell percentage of spleen cells from HBcAg- T_1T_2 group(20.21?2.01)% was higher than T_1T_2 peptides group(15.33?1.45)% and blank group(5.09?1.66)%,the percentage of IFN-?positive cells in these three groups were(1.58?0.05)%,(0.88?0.02)% and(0.53?0.03)%.The ELISA discovered that the level of IL-12 in the experimental group was the highest.Different from above,the IL-4 and IL-5 were lower in the exper- imental group.The detection of eytotoxic T lymphocyte(CTL)activity showed that the quantity of Hela cells infected by HCV in HBcAg-T1T2 stimulated group was different obviously from T1T2 peptides group. Conclusion The mixed protein HBcAg-T1 and HBcAg-T2 can induce stronger cellular immunity and it is able to serve as a therapeutic vaccines candidate specific for HCV.