C.-L. Jen

Chronic Hepatitis C Virus Infection Increases Mortality From Hepatic and Extrahepatic Diseases: A Community-Based Long-Term Prospective Study

BACKGROUND The study aimed to evaluate the risk of hepatitis C virus (HCV) infection on hepatic and extrahepatic deaths. METHODS A cohort of 23 820 adults aged 30-65 years old were enrolled during 1991-1992. The seromarkers hepatitis B surface antigen (HBsAg), anti-HCV, and serum HCV RNA levels at study entry were tested. The vital status was ascertained through computerized linkage with national death certification profiles from 1991 to 2008. RESULTS There were 19,636 HBsAg-seronegatives, including 18,541 anti-HCV seronegatives and 1095 anti-HCV seropositives. Among anti-HCV seropositives, 69.4% had detectable serum HCV RNA levels. There were 2394 deaths that occurred during an average follow-up period of 16.2 years. Compared with anti-HCV seronegatives, anti-HCV seropositives had higher mortality from both hepatic and extrahepatic diseases, showing multivariate-adjusted hazard ratio (95% confidence interval) of 1.89 (1.66-2.15) for all causes of death; 12.48 (9.34-16.66) for hepatic diseases; 1.35 (1.15-1.57) for extrahepatic diseases; 1.50 (1.10-2.03) for circulatory diseases; 2.77 (1.49-5.15) for nephritis, nephrotic syndrome, and nephrosis; 4.08 (1.38-12.08) for esophageal cancer; 4.19 (1.18-14.94) for prostate cancer; and 8.22 (1.36-49.66) for thyroid cancer. Anti-HCV seropositives with detectable HCV RNA levels had significantly higher mortality from hepatic and extrahepatic diseases than anti-HCV seropositives with undetectable HCV RNA. CONCLUSIONS Monitoring HCV RNA in anti-HCV seropositives is essential for the prediction of mortality associated with hepatitis C.

Development and validation of a clinical scoring system for predicting risk of HCC in asymptomatic individuals seropositive for anti-HCV antibodies.

Background The development of a risk assessment tool for long-term hepatocellular carcinoma risk would be helpful in identifying high-risk patients and providing information of clinical consultation. Methods The model derivation and validation cohorts consisted of 975 and 572 anti-HCV seropositives, respectively. The model included age, alanine aminotransferase (ALT), the ratio of aspirate aminotransferase to ALT, serum HCV RNA levels and cirrhosis status and HCV genotype. Two risk prediction models were developed: one was for all-anti-HCV seropositives, and the other was for anti-HCV seropositives with detectable HCV RNA. The Coxs proportional hazards models were utilized to estimate regression coefficients of HCC risk predictors to derive risk scores. The cumulative HCC risks in the validation cohort were estimated by Kaplan-Meier methods. The area under receiver operating curve (AUROC) was used to evaluate the performance of the risk models. Results All predictors were significantly associated with HCC. The summary risk scores of two models derived from the derivation cohort had predictability of HCC risk in the validation cohort. The summary risk score of the two risk prediction models clearly divided the validation cohort into three groups (p<0.001). The AUROC for predicting 5-year HCC risk in the validation cohort was satisfactory for the two models, with 0.73 and 0.70, respectively. Conclusion Scoring systems for predicting HCC risk of HCV-infected patients had good validity and discrimination capability, which may triage patients for alternative management strategies.

Association between obesity, hypertriglyceridemia and low hepatitis B viral load

Objective:This study aimed to investigate the metabolic risk factors of high hepatitis B viral load.Design:Large-scale, community-based cross-sectional study.Subjects:A total of 3587 hepatitis B virus (HBV)-infected participants without liver cirrhosis at study entry were investigated. High HBV viral load was defined as a serum level ⩾104 copies per ml for hepatitis B e antigen (HBeAg) seronegatives or ⩾108 copies per ml for HBeAg seropositives.Results:Among HBeAg seropositives (n=545), high HBV viral load was reversely associated with extreme obesity (odds ratio (OR), 0.30; 95% confidence interval (CI), 0.13–0.68; P=0.004) or central obesity (OR, 0.53; 95% CI, 0.34–0.82; P=0.004) after adjustment for gender, hypertriglyceridemia, hyperuricemia and history of hypertension. High HBV viral load remained significantly inversely associated with extreme obesity (OR, 0.17; 95% CI, 0.05–0.63; P=0.008) and central obesity (OR, 0.44; 95% CI, 0.25–0.78; P=0.005) in male HBeAg-seropositive participants in stratification analyses by gender. Among HBeAg seronegatives (n=3042), however, high HBV viral load was inversely associated with hypertriglyceridemia (OR, 0.74; 95% CI, 0.61–0.89, P=0.002) after adjustment for age, gender, high serum alanine aminotransferase level, and extreme obesity or central obesity. High HBV viral load was still inversely associated with hypertriglyceridemia in both female (OR, 0.70; 95% CI, 0.50–0.97; P=0.041) and male (OR, 0.75; 95% CI, 0.60–0.94; P=0.011) HBeAg-seronegative participants.Conclusion:Extreme obesity and central obesity were associated with a low prevalence of high HBV viral load in HBeAg seropositives, especially in men; while hypertriglyceridemia was associated with a low prevalence of high viral load in HBeAg seronegatives in both women and men.

Chronic hepatitis C virus infection and the risk for diabetes: a community-based prospective study

The association between hepatitis C virus (HCV) infection and the occurrence of type II diabetes remains controversial. Prospective studies are needed to assess its causal temporality.

Human leukocyte antigen variants and risk of hepatocellular carcinoma modified by hepatitis C virus genotypes: A genome‐wide association study

We conducted a genome‐wide association study to discover genetic variants associated with hepatitis C virus (HCV)–related hepatocellular carcinoma (HCC). We genotyped 502 HCC cases and 749 non‐HCC controls using the Axiom‐CHB genome‐wide array. After identifying single‐nucleotide polymorphism clusters located in the human leukocyte antigen (HLA) region which were potentially associated with HCC, HLA‐DQB1 genotyping was performed to analyze 994 anti‐HCV seropositives collected in the period 1991‐2013 in a community‐based cohort for evaluating long‐term predictability of HLA variants for identifying the risk of HCC. Cox proportional hazards models were used to estimate the hazard ratios and 95% confidence intervals of HLA genotypes for determining the aforementioned HCC risk. Eight single‐nucleotide polymorphisms in the proximity of HLA‐DQB1 were associated with HCC (P < 8.7 × 10−8) in the genome‐wide association study. Long‐term follow‐up showed a significant association with HLA‐DQB1*03:01 and DQB1*06:02 (P < 0.05). The adjusted hazard ratios associated with HCC were 0.45 (0.30‐0.68) and 2.11 (1.34‐3.34) for DQB1*03:01 and DQB1*06:02, respectively. After stratification by HCV genotypes, DQB1*03:01 showed protective effects only in patients with HCV genotype 1, whereas DQB1*06:02 conferred risk of HCC only in patients with HCV non‐1 genotypes. HLA imputation analyses revealed that HLA‐DRB1*15:01, which is in linkage disequilibrium with DQB1*06:02, also increased the risk of HCC (odds ratio, 1.96; 95% confidence interval, 1.31‐2.93). Haplotype analysis supported that DQB1*03:01 and DQB1*06:02 are primarily protective and susceptible variants, respectively. Conclusion: HLA‐DQB1 was independently associated with HCC; HCV genotypes modified the effects of HLA‐DQB1 on the risk of HCC. (Hepatology 2018;67:651‐661).

12 INCIDENCE AND DETERMINANTS OF SPONTANEOUS DECLINE OF HBV DNA TO UNDETECTABLE LEVEL IN PATIENTS WITH HIGH VIRAL LOAD

thus to distinguish HBV infected hepatocytes from non-infected ones. These antibodies were used to analyze the distribution and quantification of pMHC on HBV infected cells and to test their ability to deliver compounds to infected cells. Methods: Antibody secreting B cell hybridomas were generated from mice immunized with HBVc18–27/A201 and HBVe183–91/A201 pMHC monomers. The number of pMHC complexes on individual cells was enumerated by flow cytometry based quantification technique, while HBV antigen was quantified by Western blot and Quantity One Software. Confocal microscopy was used to detect intracellular delivery of fluorescent dye. Results: Culture supernatants from numerous hybridomas were screened for specificity. Five of them were able to produce antibodies that recognized T2 cells pulsed with core 18–27 (2 clones) or env 183–91 peptides (3 clones) but not T2 cells pulsed with irrelevant HLA-A0201 binding peptide. The two TCRlike antibodies could detect pMHC complexes on the surface of HepG2 cells with sensitivity lower than that of T cells (1nM versus 1pM) but they recognized the pMHC expressed naturally after endogenous processing on HBV infected cells. The binding of the antibodies was not inhibited by the presence of serum HBV antigens. The TCR-like antibodies were used to quantify pMHC complexes expressed on different cell types. On average, 1×105 core proteins produced one pMHC on B cells, while in hepatocytes, a single pMHC requires endogenous synthesis of approximately 1.5×106 proteins, directly demonstrating the limited antigen processing ability of hepatocytes. Furthermore, TCR-like antibodies could deliver the fluorescent dye to the endosomal compartment of HBV-infected hepatocytes. Conclusions: We have produced TCR-like antibodies recognizing HBV infected cells. These antibodies open the possibilities to obtain precise quantitative analysis of the peptide/HLA-class I complexes on HBV infected hepatocytes and provide the base for new therapeutic strategies to deliver drugs specifically to HBV infected cells.

722 INCIDENCE AND DETERMINANTS OF SPONTANEOUS HEPATITIS B SURFACE ANTIGEN SEROCLEARANCE: A COMMUNITY-BASED LONG-TERM FOLLOW-UP STUDY

722 INCIDENCE AND DETERMINANTS OF SPONTANEOUS HEPATITIS B SURFACE ANTIGEN SEROCLEARANCE: A COMMUNITY-BASED LONG-TERM FOLLOW-UP STUDY J. Liu, H.-I. Yang, M.-H. Lee, S.-N. Lu, C.-L. Jen, L.-Y. Wang, S.-L. You, U.H. Iloeje, C.-J. Chen, R.E.V.E.A.L.-HBV. Genomics Research Center, Academia Sinica, Taipei, Taiwan R.O.C.; Emory University Rollins School of Public Health, Atlanta, GA, USA; Graduate Institute of Epidemiology, National Taiwan University, Taipei, Department of Gastroenterology, Kaohsiung Chang-Gung Memorial Hospital, Kaohsiung, MacKay Medical College, Taipei, Taiwan R.O.C.; Research and Development, Bristol-Myers Squibb, Wallingford, CT, USA E-mail: jessica.b.liu@gmail.com

A Glycomarker for Short-term Prediction of Hepatocellular Carcinoma: A Longitudinal Study With Serial Measurements

Objectives: Wisteria floribunda agglutinin‐positive human Mac‐2‐binding protein (WFA+‐M2BP) is a glycomarker. The present community‐based long‐term follow‐up study repeatedly determined the serum WFA+‐M2BP level and examined its short‐ and long‐term associations with hepatitis C virus (HCV)‐related hepatocellular carcinoma (HCC). Methods: A total of 921 participants with antibodies against HCV seropositive, but seronegative for hepatitis B surface antigen were enrolled from seven townships in Taiwan during 1991–1992. The participants were regularly followed and their serum WFA+‐M2BP levels were measured at baseline and follow‐up. HCC was ascertained through active follow‐up and computerized data linkage with the National Cancer Registration System until December 31, 2013. Cox proportional hazards and logistic regression models were applied to estimate the magnitude of associations between serum WFA+‐M2BP levels and HCC. Results: During a median follow‐up of 21.7 years, 122 new‐onset HCC cases were identified. Elevated serum WFA+‐M2BP levels were associated with an increased risk of HCC (p < 0.001). Patients with increasing changes in serum WFA+‐M2BP levels, relative to their baseline levels, had a 4.36‐fold risk of HCC. The areas under receiver operating curves (AUROCs) of WFA+‐M2BP for predicting HCC showed that the prediction efficacy was significantly higher while closer to HCC diagnosis (p = 0.024). The AUROC was 0.91 for predicting HCC within 1 year by including the predictors of age, sex, alanine aminotransferase, alpha‐fetoprotein (AFP) and WFA+‐M2BP. Conclusions: Serum WFA+‐M2BP level may elevate before HCC onset and is a short‐term predictor of HCC among patients infected with HCV.

Association Between High Levels of Hepatitis B Core Antibody and Seroclearance of Hepatitis B e Antigen in Individuals With Chronic Hepatitis B Virus Infection

For chronic hepatitis B patients, hepatitis B e antigen (HBeAg) seroclearance signals a transition from an immunologically active phase to an inactive carrier state with a reduction in hepatitis B virus (HBV) DNA levels and a reduced risk of hepatocellular carcinoma (HCC).1 Predictors of HBeAg seroclearance include lower HBV DNA levels, viral genotype, the precore mutation, and higher serum alanine aminotransferase (ALT) levels.2.

Level of Hepatitis B (HB) Core Antibody Associates With Seroclearance of HBV DNA and HB Surface Antigen in HB e Antigen-Seronegative Patients

BACKGROUND & AIMS: Although a low level of hepatitis B surface antigen (HBsAg) is a marker of hepatitis B virus (HBV) seroclearance, additional biomarkers are needed for more accurate prediction. We investigated whether quantification of antibody against HBV core protein (anti‐HBc) can identify patients with undetectable levels of HBV DNA and HBsAg seroclearance among those who were HBV e antigen (HBeAg)‐seronegative. METHODS: We performed a retrospective analysis of data from a community‐based cohort of individuals (30–65 years old) in Taiwan who were HBsAg seropositive, anti‐HCV negative, and free of cirrhosis and/or liver cancer, recruited from 1991 through 1992, and evaluated every 6–12 months until June 30, 2004. We measured levels of anti‐HBc in blood samples from 2500 participants who were seronegative for HBeAg. The first date at which a sample tested negative for HBV DNA or HBsAg, and remained negative in subsequent tests, was designated as the date of spontaneous HBV DNA undetectability or HBsAg seroclearance. We calculated cumulative incidences of HBV DNA undetectability and HBsAg seroclearance; associations between level of anti‐HBc and undetectability of HBV DNA or HBsAg seroclearance were estimated by Cox proportional hazard regression. The effects of time on the associations between level of anti‐HBc and HBsAg seroclearance was assessed by the area under the receiver operating characteristic curve (AUROC) analysis. RESULTS: After a 12‐year follow‐up period, higher proportions of subjects with levels of anti‐HBc <3 log IU/mL had undetectable levels of HBV DNA (58%) and HBsAg seroclearance (53%) than subjects with higher levels of anti‐HBc (29.6% and 19.8%, respectively) (P < .001). For subjects with levels of HBsAg <102 IU/mL and anti‐HBc <3 log IU/mL, the adjusted rate ratio of HBV DNA undetectability was 16.45 (95% CI, 11.15–24.28) and of HBsAg seroclearance was 17.95 (95% CI, 12.49–25.81), compared to subjects with higher levels of HBsAg and anti‐HBc. A model that included level of anti‐HBc as a parameter identified subjects with HBsAg seroclearance within 10 years with an AUROC of 82%; this value was significantly higher than that from models that include only level of HBV DNA and HBsAg (P < .0001). CONCLUSIONS: In a retrospective analysis of a large cohort of patients with chronic HBV infection in Taiwan, we associated levels of anti‐HBc <3 log IU/mL with undetectable HBV DNA and HBsAg seroclearance occurred within 10 years; patients who also have levels of HBsAg <102 IU/mL have greater odds. Combining data on levels of HBsAg, HBV DNA, and anti‐HBc is able to identify HBeAg‐seronegative patients who can achieve HBsAg seroclearance with an AUROC value of 82%.