11/15/2005 8:00 AM - 12:30 PM

Hepatitis B Pathogenesis

 

 

 

Poster 1285

Abstract ID: 62098

Category: JO9: Hepatitis B: Pathogenesis

Significance of 'anti-HBc only' in unselected sera referred to the clinical laboratory.

J. Masurel, Université Paris-Descartes, Hôpital Cochin, Service de Virologie, Paris,

France, O. Launay, Université Paris-Descartes, CIC de Vaccinologie Cochin-Pasteur,

Paris, France, A. Basse-Guérineau, AP-HP, Hôpital Cochin, Service de Virologie, Paris,

France, A. Servant-Delmas, INTS, Centre National de Référence des Hépatites B et C en

Transfusion, Paris, France, J. Méritet, AP-HP, Hôpital Cochin, Service de Virologie,

Paris, France, S. Laperche, INTS, Centre National de Référence des Hépatites B et C en

Transfusion, Paris, France, P. Sogni, Université Paris-Descartes, Hôpital Cochin, Service d'Hépatologie, Paris, France, A. R. Rosenberg, Université Paris-Descartes, Hôpital Cochin, Service de Virologie, Paris, France

 

Background & Aim:

Isolated detection of total anti-HBc in the absence of HBsAg and anti-HBs (‘anti-HBc only’) has been reported, particularly among patients infected with human immunodeficiency virus (HIV) or hepatitis C virus (HCV). It remains unclear how this serological profile should be interpreted and whether all individuals with such pattern need to be tested for hepatitis B virus (HBV) DNA. The aim of this study was to evaluate the significance of ‘anti-HBc only’ in unselected sera.

 

Methods & Results:

Of the 6431 patients that were tested for HbsAg, anti-HBs, and total anti-HBc in the clinical laboratory of our teaching hospital between January and

December 2003, 362 (5.6%) had ‘anti-HBc only’. Among them, 1 was in the window period following acute hepatitis B (positive for anti-HBc IgM). Anti-HBe was undetectable in 207 (57.2%) sera, suggesting that some of these might be false positive. However, only 11 of these 207 sera were negative when retested using another assay for total anti-HBc; these 11 sera had significantly lower sample/cutoff ratios when compared to those positive with both assays (p<0.001). Two sera had detectable HBsAg when retested with one of the most recent HBsAg assays. In these 2 sera, HBV DNA was

detected, and sequencing of the s gene revealed amino acid substitutions in the immunodominant epitope of HBsAg. The remaining 348 sera with true ‘anti-HBc only’ were tested for HBV DNA using the branched DNA-based VERSANT HBV DNA 3.0 assay (Bayer), a standardized test with a threshold of detection (2,000 copies/ml) that might allow the screening of clinically relevant infection. The PCR-based COBAS AMPLICOR HBV MONITOR assay (Roche) was used as confirmation test. HBV DNA was found in 8/348 (2.3%) patients, including 1 pregnant woman, with viral loads above 10,000 copies/ml in 5 cases. No mutation was detected in the s gene in these 8 cases. No factor (including HIV and HCV status) was found to be predictive of HBV DNA positivity.

 

Conclusions:

Despite the use of more sensitive HBsAg assays, ‘anti-HBc only’ is a common event in the clinical laboratory, and is rarely due to false positivity. In view of the absence of predictors of chronic HBV infection, the results suggest that all individuals with ‘anti-HBc only’ should be tested for serum HBV DNA in order to identify patients at potential risk of HBV transmission, and needing clinical and virological follow-up.

 

 


Poster 1286

Abstract ID: 63416

Category: JO9: Hepatitis B: Pathogenesis

Etiopathology of HCC in Thailand: Implication of occult HBV infection and correlation with p53 and &#946;-catenin mutations.

O. Galy, INSERM U271, LYON, France, Metropolitan, I. Chemin, INSERM U271,

LYON CEDEX 03, France, S. Villar, IARC Unit of nutrition and Cancer, LYON,

France, M. Chevallier, Pathology Laboratory, Marcel Merieux Laboratory, LYON,

France, P. Srivatanakul, National Cancer Institute Cancer Control Unit, Bangkok,

Thailand, C. Trépo, INSERM U271, Lyon, France, P. Hainault, IARC Molecular

Carcinogenesis Unit, LYON , France

 

Introduction and Aim:

Hepatocellular Carcinoma is the main primary liver cancer observed in the world representing more than 70% of the 500 000 annual new cases of liver cancer. The major risk factor is Hepatitis B virus (HBV) chronic infection that increases cancer-risk about 100 times. The existence of HBV occult infection (HBV DNA without HBsAg) is now recognized but its role in liver disease, especially cirrhosis and HCC is not clearly elucidated. Many genetic alterations were identified in HCC; pathway involved in growth control, apoptosis or DNA repair. However, the molecular basis of how they interfere with HBV and whether HBV occult infections follow the same pathways in the pathogenesis of HCC is still poorly understood.

 

Methods:

In this study we describe TP53 and &#946;-catenin mutations according to HBV infection status (occult or HBsAg+) in a high HVB prevalence zone (Thailand). 33 HCC cases were identified among 50 pairs of liver cancer tumours selected from a large study conducted in Thailand (one pair including tumour tissue and a respective surrounding non-tumorous area). Viral status of samples (HBV and Hepatitis C Virus (HCV)) was determined by PCR and RT-PCR respectively, we eventually selected 18 “HBV DNA+”

pairs. 7 out of the 18 pairs exhibited HBsAg negative status; they were categorized as “occult HBV” pairs. HCV RNA was detected in 3 out of the 18 pairs including one case with an occult HBV pair. This low incidence of HCV infection excludes hypothesis of silent hepatitis due to HCV repression effect. Determination of tumour grade (Edmondson) and Fibrosis/Activity (METAVIR) scores for surrounding area were performed. We identified TP53 gene mutations by PCR/RFLP/Sequencing in exon 5-9 and &#946;-catenin mutations by DHPLC/Sequencing in exon 3.

 

Results:

Age of patients positive for HBV DNA ranged from 34 to 73 with an average value of 48, age of patients with occult hepatitis was lower than with HBsAg+ Hepatitis. Tumour grade and METAVIR scores for non tumour areas were similar in the two types of infection. TP53 mutations were detected in 7 cases; aspecific TP53 mutation at codon 249 represented 60% of those mutations but only occurred among HBsAg+ cases. &#946;-catenin mutations distribution was similar for occult and non-occult infection.

 

Conclusion:

The study suggests that there is no significant disparity between the distributions of different molecular events in the two HBV infection types. Thus we can conclude that HBV occult infection could be implicated in HCC development according to mechanism similar to those involved in non-occult infection. The possible association of occult infection with TP53 mutation at codon 249 requires more detailed investigation.

 

 


Poster 1289

Abstract ID: 65055

Category: JO9: Hepatitis B: Pathogenesis

Adefovir treatment of chronic hepatitis B patients recovers circulating numbers and function of myeloid dendritic cells.

R. G. van der Molen, Erasmus MC, University Medical Center Rotterdam, Rotterdam,

Netherlands, D. Sprengers, Erasmus MC, University Medical Center Rotterdam,

Rotterdam, Netherlands, P. J. Biesta, Erasmus MC, University Medical Center

Rotterdam, Rotterdam, Netherlands, R. A. de Man, Erasmus MC, University Medical

Center Rotterdam, Rotterdam, Netherlands, H. G. Niesters, Erasmus MC, University

Medical Center Rotterdam, Rotterdam, Netherlands, J. G. Kusters, Erasmus MC,

University Medical Center Rotterdam, Rotterdam, Netherlands, H. L. Janssen, Erasmus

MC, University Medical Center Rotterdam, Rotterdam, Netherlands

 

Introduction:

Dendritic cells (DC) play an important role in the induction of T-cell responses. The myeloid DC (mDC) and plasmacytoid DC (pDC) are the two predominant precursor DC-subtypes present in peripheral blood. Recently, we have shown that mDC of chronic hepatitis B virus (HBV) infected patients are impaired in the expression of costimulatory molecules, T-cell stimulatory capacity as well as TNFa production. In addition, we showed a reduced IFNa production by pDC in chronic HBV patients compared to healthy controls. The question is whether the impaired DC function contributes to chronicity of HBV infection or whether the virus itself causes this dysfunction of DC.

 

Aim:

The aim of our study was to assess the effect of virus reduction by adefovir on function and absolute numbers of both mDC and pDC.

 

Methods:

In this study, 14 chronic HBV patients were treated with adefovir (10 mg od.) and followed during the first 6 months of treatment.

 

Results:

The median serum HBV DNA decreased significantly from 1.1x108 to 2.0x103 geq/ml within the first 3 months and remained stable thereafter. The mean ALT levels decreased from 150 to 41 U/L after 6 months. As determined by flowcytometry, a significant increase in the absolute numbers of mDC, but not of pDC was found after 6 months of treatment compared to baseline. To determine DC function, mDC and pDC were isolated using magnetic cell sorting techniques. The T-cell stimulatory capacity of mDC increased significantly in 10 out of 14 patients after 3 months of treatment. The mean IL-12 as well as the TNFa production by mDC stimulated with poly (I:C) and IFNg significantly increased after 3 months of treatment in these 10 patients. In 4 out of 14 patients, the T-cell stimulatory capacity decreased after 3 months compared to baseline. At the same time, IL- 12 and TNFa levels were reduced. Two of these patients started treatment at time of immune activation, i.e. ALT elevated 8 and 27 times the upper limit of normal. This indicates that their mDC might have been activated at baseline. The longitudinal expression of costimulatory molecules CD80, CD86 and CD40 concurred with T-cell stimulation results for all patients. The mean production of IFNa by isolated pDC stimulated with Staphylococcus aureus Cowan strain I antigen showed a trend towards reduction after 6 months of treatment compared to baseline.

 

Conclusion:

In conclusion, reducing the viral load by adefovir recovers circulating numbers and function of mDC, but not of pDC, suggesting that adefovir contributes to partial restoration of the immunological control to HBV infection.

 


Poster 1294

Abstract ID: 64494

Category: JO9: Hepatitis B: Pathogenesis

The Relationship between Hepatitis B Virus (HBV) Genotype and Hepatitis B e Antigen (HBeAg) Status in a Cohort of Alaska Natives.

S. Livingston, Liver Disease and Hepatitis Program, Anchorage, AK, J. Simonetti, Liver

Disease and Hepatitis Program, Anchorage, AK, L. Bulkow, Arctic Investigations

Program, CDC, Anchorage, AK, C. Homan, Liver Disease and Hepatitis Program,

Anchorage, AK, M. Snowball, Liver Disease and Hepatitis Program, Anchorage, AK, K.

Hurlburt, Liver Disease and Hepatitis Program, Anchorage, AK, J. Williams, Liver

Disease and Hepatitis Program, Anchorage, AK, B. McMahon, Liver Disease and

Hepatitis Program, Anchorage, AK

 

Introduction:

In Alaska Natives (AN) with chronic HBV infection, we have previously observed a 73% probability of clearing HBeAg within 10 years of the first positive hepatitis B surface antigen (HBsAg) specimen. Objective: To investigate the relationship between HBV genotype (GT) and HBeAg status in this cohort.

 

Methods:

Followed 1536 AN with chronic HBV infection for 12.6 years (median).

Sera were tested biannually for HBeAg. Using stored sera, we tested for HBV GT in 1045 persons by direct sequencing of the S gene and determined HBeAg status by GT on the initial HBsAg + specimen and changes in HBeAg status by GT over time.

 

Results:

Genotypes A, B, C, D, F and H (1 case--omitted from further analysis) were

identified. Genotype distribution and proportion HBeAg+ by GT is displayed in the Table. The median age (years) at time of initial HBeAg+ specimen by GT was A, 15.1; B, 18.9; C, 19.6; D, 12.8, and F, 11.3. Fewer persons with GT A and B were initially HBeAg+ (P<.001); this remained true after adjustment for age (P<.001). Among persons initially HBeAg+, those with GT C were significantly more likely to remain positive on subsequent visits than those with GT A and D (P<.001), who likewise were more likely to remain positive than those with GT B and F (P<.001). At time of last testing, more persons with GT C, 71% (22/31), were still HBeAg positive compared to those with GT A, B, D and F, 14% (4/28), 0% (0/5) , 41% (63/255) and 13% (14/109) respectively (P<.001). Median age at seroconversion is shown in the Table. Persons with GT C were significantly older at the time of HBeAg seroconversion (P<.001). Among those initially

HBeAg+, persons with GT C and F were more likely to revert to HBeAg+ after clearing HBeAg than those with other GT (P<.001). Of persons initially HBeAg-, those with GT B, D and F were more likely to remain HBeAg- than GT A and C (P=.002).

 

Conclusions:

Significant differences in HBeAg seroconversion and reversion back

to HBeAg positivity were observed between HBV GT among a homogeneous population of AN. Persons with GT C were significantly less likely to clear HBeAg and significantly more likely to have reversions back to HBeAg positivity than those with other GT, suggesting that HBV GT status may be of clinical importance.

 

 


Poster 1306

Abstract ID: 66121

Category: JO9: Hepatitis B: Pathogenesis

Adefovir treatment of chronic hepatitis B decreases regulatory T cells and partially restores the antiviral immune response.

J. N. Stoop, Erasmus MC, University Medical Center Rotterdam, Rotterdam,

Netherlands, R. G. Van der Molen, Erasmus MC, University Medical Center Rotterdam,

Rotterdam, Netherlands, E. J. Kuipers, Erasmus MC, University Medical Center

Rotterdam, Rotterdam, Netherlands, R. A. de Man, Erasmus MC, University Medical

Center Rotterdam, Rotterdam, Netherlands, J. G. Kusters, Erasmus MC, University

Medical Center Rotterdam, Rotterdam, Netherlands, H. L. Janssen, Erasmus MC,

University Medical Center Rotterdam, Rotterdam, Netherlands

Disclosures: The following authors have indicated they have no relationships to disclose:

Jeroen Stoop, Renate Van der Molen, Ernst Kuipers, Robert de Man, Johannes Kusters,

Harry Janssen

 

Introduction:

Adefovir is a nucleotide analogue, which effectively suppresses viral replication in the majority of chronic HBV infected patients. We determined whether adefovir can induce a restoration of the immune response in patients with chronic hepatitis B. An important mediator of impaired immune reactivity are regulatory T cells (Treg). We previously showed that Treg are capable of inhibiting the immune response against HBV core antigen (HBcAg).

 

Methods:

In the current study we assessed the effect of adefovir therapy on the percentage of Treg and the immune response against HBcAg. Peripheral blood mononuclear cells (PBMC) from 12 chronic hepatitis B patients were obtained at baseline and after 3 and 6 months of adefovir treatment (10 mg per os daily). All patients had serum HBVDNA levels above 105 copies/ml and abnormal ALT levels. Six patients were HBeAg positive and 6 anti-HBeAg positive. Samples from one patient were tested in the same experiment. The percentage of Treg of CD4+ cells was determined by flowcytometry with antibodies against CD4, CD25, CD45RO and CTLA-4. The response against HBV was tested by stimulating PBMC with HBcAg. After stimulation the IFN-g and IL10 production were determined by Elispot. The proliferation was determined by 3[H]-thymidine incorporation.

 

Results:

The median drop of HBVDNA was 5.16 log after 3 months and HBVDNA levels remained stable thereafter. Immunological testing has been performed for 7 patients so far. The percentage Treg of CD4+ cells at baseline was 3.60 ± 0.44 (mean ± sem) and decreased to 2.81 ± 0.22 at 3 months (P=0.028) During month 3-6 the percentage of Treg did not change significantly. During the first 3 months of treatment the HBcAg specific proliferation increased from 3736 CPM ± 984 to 5537 CPM ± 1350 and the IFN-g production increased from 7.05 spots/100.000 cells ± 2.61 to 21.48 spots/100.000 cells ± 7.96. After these 3 months the proliferation and the IFN-g production did not increase any further. Treatment with adefovir did not affect the HBcAg specific IL10 production.

 

Conclusion:

In conclusion, during the treatment with adefovir the proportion of Treg in

peripheral blood decreases while both the IFN-g production and proliferation

against HBcAg increases. However, we did not find a decrease in the production of the anti-inflammatory cytokine IL-10. Together these data suggest that adefovir contributes to a partial recovery of the immunological control of chronic HBV infection.


Poster 1308

Abstract ID: 67083

Category: JO9: Hepatitis B: Pathogenesis

Models To Predict Hepatocellular Carcinoma (HCC) In Patients With Chronic Hepatitis B Infection: The R.E.V.E.A.L.- HBV Study.

C. Chen, National Taiwan University, Taipei , Taiwan, H. Yang, National Taiwan

University, Taipei , Taiwan, U. H. Iloeje, Bristol-Myers Squibb Pharmaceutical

Research Institute, Wallingford, CT, J. Su, Bristol-Myers Squibb Pharmaceutical

Research Institute, Wallingford, CT, C. Jen, National Taiwan University, Taipei ,

Taiwan, S. You, National Taiwan University, Taipei , Taiwan, Y. Liaw, Chang Gung

Memorial Hospital and University, Taipei , Taiwan

 

Introduction:

The risk of any particular CHB patient developing HCC over a specific

period remains to be determined. Predicting patients at risk of progressing to HCC may help in patient counseling, risk stratification and identifying those most likely to benefit from intervention. Our objective was to develop such a model using readily available clinical information in CHB infected subjects.

 

Methods:

Information from 3,653 asymptomatic HBsAg-positive and anti-HCV-negative subjects recruited from 7 townships in Taiwan between 1991 and 1992 was used for these analyses. The subjects were randomized into two groups (training and validation) on a 1:1 basis. HCC cases were ascertained as part of ongoing analyses by computerized data linkage to the Taiwanese National Cancer Registry and Death Certificates Profiles or by ultrasound

screening. In addition, the medical charts of these subjects were reviewed using a structured abstraction form. A diagnosis of HCC was confirmed by any one of the following: positive cyto-histopathology; presence of two coincident, focal liver lesions by liver imaging studies (abdominal ultrasonography, CT Scan, MRI and/or angiography); presence of one focal liver lesion and an alpha fetoprotein (AFP) level >400 ng/ml. Logistic regression models were used to identify the predictive variables, and the model accuracy determined by the Area Under the Receiver Operator Characteristic curves (AUROC).

 

Results:

The mean follow-up time was 11 years in this cohort. The training set 1826/3851 (50%), was comparable to the validation set for all baseline variables and HCC events (p>0.05). Distribution of key variables in both groups were, males (62%); HBeAg-positive (16%); ALT <1x ULN (94%); current alcohol consumption (12%). Using multivariable logistic regression, in the best fitting model Age and HBV DNA level were the strongest predictors of future HCC events (AUROC = 0.87); concordant percent 95%; discordant percent 5%; model goodness of fit (p=0.33). The strongest risk

was associated with age 60-65; OR (95% CI) was 16.1 (6.3-41.4) reference group (30- 39); and HBV DNA ³ 105 copies/mL OR (95% CI) was 9.3 (4.0-21.4) [reference <300 copies/mL]. The model had a sensitivity of 78% and specificity of 78% at a predictive probability level of 4.3%.

 

 

Model characteristics in the validation group were AUROC = 0.83; concordant percent =95.6; and goodness of fit =0.3008.

 

Conclusion:

Risk stratification for HCC is possible using a model comprised of readily available non-invasive clinical information. External validation of the model will be necessary to further evaluate its characteristics.

 

 


Poster 1309

Abstract ID: 67282

Category: JO9: Hepatitis B: Pathogenesis

Serum HBV DNA levels and Liver Disease progression in patients with chronic HbeAg (-) Hepatitis B infection. A prospective cohort study.

G. Zacharakis, University of Athens, Hippokration General Hospital, Athens, Greece, I.

Koskinas, University of Athens, Hippokration General Hospital of Athens, Athens,

Greece, S. Kotsiou, Dimokrition University of Thrace, Alexandroupolis, Greece, M.

Papoutselis, Unit of Preventive Medicine, Alexandroupolis, Greece, F. Tzara, Unit of

Preventive Medicine, Alexandroupolis, Greece, N. Vafeiadis, Unit of Preventive

Medicine, Alexandroupolis, Greece, A. Archimandritis, University of Athens,

Hippokration General Hospital, Athens, Greece, K. Papoutselis, Unit of Preventive

Medicine, Alexandroupolis, Greece

 

Background:

The role of serum HBV-DNA levels in the natural history and management of chronic HBV infection is currently under investigation.

 

Aim:

To evaluate the levels of serum HBV-DNA in association with the natural

history of chronic HBV infection. A prospective cohort study sponsored by a grant from EC Project Interreg I and II.

 

Patients/Methods:

257 adults, median age 34 years (range 20-65), with chronic HBV infection, were recruited during 1992-1994 and prospectively followed up for a period up to 12 years (median 5.3 years). Patients with HCV/HDV coinfection, alcohol consumption or treated were excluded from the study. Viral markers, liver biochemistry and physical examination were performed every 6 months. Liver biopsy was done at entry and every 2-4 years. Liver histology was assessed according to Scheuer criteria. Serum HBV-DNA levels were measured by Amplicor HBV Monitor kit (Roche Diagnostics Systems, Switzerland).

 

Results:

14/257 (5%) patients were HBeAg(+) with median serum HBV DNA

levels 6.9x106 copies/ml (range 1.4x105-4x107copies/ml) At entry, 195/257 (76%) were HBeAg (-), anti-HBe (+), with a history of normal ALT levels for at least 6 months (inactive carries): a) 97/195(50%) individuals had undetectable serum HBV-DNA, no liver disease and remained so at follow-up period, b) 94/195 (48%) subjects had serum HBV-DNA levels <104 copies/ml, 91/94 (97%) normal liver histology and remained so at follow-up period except 6 patients who lost serum HBV-DNA, and, c) 4/195 (2%) individuals had serum HBV-DNA >104copies/ml, nearly normal liver histology of whom 2 exhibited abnormal ALT levels after 3 years and developed mild progression of liver disease. At entry, 48/257 (19%) patients were HBeAg (-), anti-HBe (+), with a history of elevated ALT levels for at least 6 months; 12/48 (25%) had an erratic ALT pattern, median HBV-DNA levels 3.5x105copies/ml (range 7.1x104 to 3.2x107) and 36/48 (75%) had persistent elevated ALT(>1.5xUNL), median HBV-DNA levels 1.2x107copies/ml (range 2x105 to 6.9x107), (p<0.05). In this group of patients, 22/48 (46%) had moderate/severe histology at entry and 5/48 (10%)

developed liver disease progression during the follow-up period. Overall,

HBeAg(-) patients with persistent serum HBV-DNA levels >104copies/ml were more likely to have liver disease progression than those with lower or

undetectable HBV-DNA levels (OR: 2.1).

 

Conclusions:

The threshold of serum HBV-DNA associated with abnormalities

in liver biochemistry and liver disease seems to be the level of >104copies/ml. Close monitoring of serum HBV-DNA could be a useful clinical tool in the management of chronic HBV patients, particularly in the HBeAg (-) group.

 


Poster 1310

Abstract ID: 61212

Category: JO9: Hepatitis B: Pathogenesis

Non-invasive assessment of liver fibrosis in chronic hepatitis B using FibroScan®.

P. Marcellin, Hopital Beaujon, Clichy, France, Metropolitan, V. De Ledinghen, Hopital

Haut Leveque, Pessac, France, Metropolitan, D. Dhumeaux, Hopital Henri Mondor,

Créteil, France, Metropolitan, R. Poupon, Hopital Saint Antoine, Paris, France,

Metropolitan, M. Ziol, Hopital Jean Verdier, Bondy Cedex, France, Metropolitan, P.

Bedossa, Hopital Beaujon, Clichy, France, Metropolitan, M. Beaugrand, Hopital Jean

Verdier, Bondy Cedex, France, Metropolitan

 

Introduction:

Previous studies have shown that liver stiffness measurement (LSM) with the FibroScan® (Echosens®, Paris, France) is correlated to hepatic fibrosis in chronic hepatitis C.

 

Aim:

The aim of this prospective multi centre study was to evaluate the accuracy of this non-invasive method to assess fibrosis in patients with chronic hepatitis B.

 

Methods:

220 patients with chronic hepatitis B who underwent a liver biopsy during their routine follow-up were included in the study after giving their informed consent. LSM was performed within six months of the liver biopsy. Only LSM with at least 8 valid measurements and an IQR/median ratio >50% were considered interpretable. Fibrosis and activity were evaluated on the histological slides by two pathologists using the METAVIR and Ishak scoring systems. Steatosis was graded as follow: 0 (no steatosis), 1

(1-10%), 2 (11-30%) and 3 (>30%). Only liver samples with at least 10 portal tracts or obvious cirrhosis were considered suitable for evaluation.

38 (17%) biopsy samples were considered unsuitable for a reliable staging of fibrosis. LSM failed in 12 (5%) patients. In the 170 remaining patients, LSM ranged from 2.5 to 72.5 kPa. The distribution of METAVIR fibrosis scores was: F0 (6%), F1 (36%), F2 (30%), F3 (14%) and F4 (14%). The distribution of METAVIR activity scores was: A0 (25%), A1 (50%), A2 (18%) and A3 (7%). As well, the distribution of steatosis scores was 0 (50%), 2 (34%), 2 (8%) and 3 (8%). The spearman correlation coefficients between LSM and METAVIR or Ishak fibrosis were 0.66 and 0.67 respectively (p < 0.0001).

 

Results:

Multivariate analysis of LSM versus METAVIR activity and fibrosis and steatosis scores showed that only fibrosis stage was significantly associated with LSM. The areas under the ROC curves (95% confidence interval) were 0.81 (0.74-0.86), 0.92 (0.86-0.95) and 0.90 (0.81-0.95) for METAVIR F >= 2, F >= 3 and F=4, respectively.

 

Conclusion:

In patients with chronic hepatitis B, the accuracy (ranging from 0.81 to 0.92) of LSM to measure fibrosis stage and to detect presence of cirrhosis was not significantly different from that obtained in patients with chronic hepatitis C.

 


Poster 1312

Abstract ID: 65486

Category: JO9: Hepatitis B: Pathogenesis

Prevalence and clinical role of occult hepatitis B in a cohort of HIV-infected patients.

N. Marino, S.M.Annunziata Hospital, Firenze, Italy, S. Lo Caputo, S.M.Annunziata

Hospital, Firenze, Italy, P. Pierotti, S.M.Annunziata Hospital, Firenze, Italy, C. Blè,

S.M.Annunziata Hospital, Firenze, Italy, M. Riccardi, Misericordia Hospital, Grosseto,

Italy, M. Trezzi, Misericordia Hospital, Grosseto, Italy, M. Toti, Misericordia Hospital,

Grosseto, Italy, M. De Gennaro, Campo di Marte Hospital, Lucca, Italy, A. Scasso,

Campo di Marte Hospital, Lucca, Italy, A. Vivarelli, Del Ceppo Hospital, Pistoia, Italy,

D. Dionisio, Del Ceppo Hospital, Pistoia, Italy, F. Mazzotta, S.M.Annunziata Hospital,

Firenze, Italy

 

Background –

Hepatitis B virus is not completely eliminated after loss of hepatitis B surface antigen (HBsAg) and clinical recovery: HBV-DNA can be detected by polymerase chain reaction (PCR) assay in serum, liver and peripheral blood mononuclears cells. The HBV detection rate is highest in subjects who are anti-HBc positive/antiHBs negative (isolated anti-HBc ). Occult hepatitis B virus infection was observed in approximately 10% of HIV-infected patients with HBc alone. The screening of this subset of HIV+ patients may have clinical implications.

 

Methods-

Consecutive HIV+ patients in the year 2003 were chequed in 4 Infectious

Disease Units in Tuscany (Italy) in order to assess HIV-related parameters (CD4 count, HIV-RNA), serological markers for HBV (HBsAg, anti-HBs, anti-HBc , HBeAg, anti- HBe) and HCV (anti-HCV, HCV-RNA ). PCR for HBV-DNA (Cobas Amplicor HBV Monitor; threshold of detection: 200 HBV-DNA copies/mL) was performed in patients who showed only anti-HBc reactivity. Epidemiological, clinical and therapeutical data were collected. T.Student test was employed.

 

Results-

A total of 955 HIV-positive ( 71,3% males, median age 42,3 years, heterosexual transmission 26,7%, homosexual 30,2%, IVD 59,4%) were evaluated for HBV infection. Based on hepatic serology 581 patients (60,8%) were anti-HBc positive, 64 (6,7%) HbsAg positive, 361 (37,8%) anti-HBs positive, 190 (19,9%) anti-HBc positive alone. 402 patients (42,1%) were coinfected with HCV. Isolated antiHBc+ was not related to

sex, age, immunological status and HIV-viremia. . Patients HCV+ were more likely to have isolated anti-HBc than were subjects with HIV alone ( 33,6% vs. 9,9% p<0,0001). HBV-DNA was positive in 6,9% of anti-HBc + patients. with low level of replication (from 102 to 103 copies/mL). Patients HBV-DNA positive didn’t show either clinical or laboratory abnormalities, no evidence of relation with CD4 count, HIV-RNA, HAART, or HCV coinfection. Data on the follow-up (6-12-18 months) ) report that viraemia in

occult HBV infection tends to appear intermittently with small fluctuation of very low levels of HBV replication. 17,8% of anti-HBc+ patients was HBVDNA positive in at least 1 determination. During the follow up period a patient who has interrupted voluntarily the HIV therapy, including Lamivudine, has developed an acute hepatitis B.

 

Conclusions-

The serological pattern of isolated anti-HBc positivity is frequent in HIV infected patients: longitudinal evaluation of HBV-DNA is necessary for correct diagnosis of occult HBV infection. The screening of subjects with occult hepatitis B infection could have implications in terms of surveillance and therapy in the HIV infection.

 


Poster 1316

Abstract ID: 64208

Category: JO9: Hepatitis B: Pathogenesis

Host genetic and virus factors contribute to the clinical phenotype of hepatitis B: a twin study.

B. Xu, Institute of Infectious Diseases of PLA, Southwest Hospital,, Chongqing , China,

Y. Wang, Institute of Infectious Diseases of PLA, Southwest Hospital,, Chongqing ,

China, G. Deng, Institute of Infectious Diseases of PLA, Southwest Hospital,,

Chongqing, China, L. Lan, Institute of Infectious Diseases of PLA, Southwest Hospital,,

Chongqing , China, Y. Huang, Institute of Infectious Diseases of PLA, Southwest

Hospital,, Chongqing , China

 

Aim:

To analyze the effect of host genetic and virus factors on clinical phenotype of hepatitis B between HBV infected twins (20 pairs) and control groups (14 pairs of sex and age matched patients). Twins' zygosity was identified by short tandem repeat genescan. Hepatitis A, B, C, delta and E virus serological markers, HBV DNA level, ALT, AST and TBil were detected.

 

Methods:

The data was analyzed by Fisher's exact test. 6 patients with high

HBV DNA level were divided into 3 groups: monozygotic (MZ) twins, dizygotic (DZ) twins and non-twin siblings groups. The relationship of viral quasispecies, genotype and clinical phenotype was analyzed. The procedure was PCR-RFLP analysis of HBV S gene (HBV genotyping)-longer PCR of HBV 3.2kb genome-TA cloning-conformation sensitive gel electrophoresis analysis (quasispecies detection of preS1/S2 region)- sequencing.

 

Results:

The results were as follows: 1. 14 pairs of MZ twins and 6 pairs of DZ twins

were identified. 2. As to the concordant rate, diseases phenotype concordant rate and serological pattern of HBV infection, MZ twins had significant difference compared with DZ twins and control groups (P<0.05), but DZ twins and control groups had no significant difference (P>0.05). In all groups, no significant difference was observed in infection rate, HBsAg (+) rate, HBeAg (+) rate, asymptomatic carrier rate and HBV clearance rate (P>0.05). 2 pairs of MZ twins had distinct outcomes: one died of severe

hepatitis, the other suffered from chronic infection. 3 pairs of MZ twins changed from symptomless to different outcomes with various antiviral effects in their twenties. HBIg injection and inoculation right after birth might prevent high-risk twins from infection. 3. 1 pair of MZ twins infected with different HBV genotype (B and C) showed discordant phenotype, while 1 pair of DZ twins (genotype B) and 1 pair of siblings (genotype C) infected with similar HBV strains (genetic diversity=0.0%~3.3%) showed concordant

phenotype. 4. The long fragment gene deletions in preS1/S2 region of HBV genotype C within the siblings pair were found.

 

Conclusions:

The high concordance in MZ twins indicated that the host genetic background might influence the clinical phenotype, while other factors such

as age, antiviral and inoculation might also effect on clinical phenotype. The long fragment gene deletions in HBV preS1/S2 region might be a reason for HBsAg antigenicity change and lead to immune escape and onset of illness. In a word, the results suggest both host and virus factors can contribute to determining the outcome of hepatitis B, and further research is needed to decide which factor plays a dominant role.

[Key words] Hepatitis B virus; Twins; Genetics; Phenotype
Poster 1317

Abstract ID: 64481

Category: JO9: Hepatitis B: Pathogenesis

Clinical Significance of Hepatitis B Virus Genotypes in Children and Adolescents with Chronic Hepatitis B.

S. Wirth, Children's Hospital, HELIOS KLinikum Wuppertal, Witten-Herdecke Univ.,

Wuppertal, Germany, P. Oommen, Children's Hospital, HELIOS KLinikum Wuppertal,

Witten-Herdecke Univ., Wuppertal, Germany, P. Gerner, Children's Hospital, HELIOS

KLinikum Wuppertal, Witten-Herdecke Univ., Wuppertal, Germany, P. Wintermeyer,

Children's Hospital, HELIOS KLinikum Wuppertal, Witten-Herdecke Univ., Wuppertal,

Germany, A. Jenke, Children's Hospital, HELIOS KLinikum Wuppertal, Witten-

Herdecke Univ., Wuppertal, Germany, E. Coci, Children's Hospital, HELIOS KLinikum

Wuppertal, Witten-Herdecke Univ., Wuppertal, Germany

 

Background/Aims:

Until today the prevalence of different HBV genotypes and their

clinical significance have only rarely been investigated in children with chronic hepatitis B. Thus, the aim of this study was both to ascertain epidemiological data on HBV genotype distribution in childhood and to correlate the results with serological, virological, and histological data in order to provide information on the therapeutic and prognostic significance of genotypes in this age group.

 

Methods:

Hepatitis B virus genomes of 249 HBeAg-positive chronic HBV carriers were genotyped based on restriction fragment length polymorphism methodology (RFLP). Genotypes were correlated with corresponding values for alanine aminotransferase (ALT) levels, quantitative HBV DNA and histological findings.

 

Results:

162 males and 87 females at a mean age of 7.2 years were studied. 96 % of the patients could be attributed to HBV genotypes A and D (32.5 % and 63.5 %, respectively) wheras the remaining were classified as genotypes B, C, E and F. ALTlevels and HBV DNA-levels were available for 237 and 176 patients, respectively. There was no significant difference in ALT levels among different genotypes (mean 67 U/l). Similarly, no significant genotype-difference was found for histological findings. In contrast, there was a clear association between very high hepatitis B virus DNA levels

and individuals with HBV genotype D (p=0.006). Mean time follow-up of 3.6 years showed that patients with genotype D tended to seroconvert to anti-HBe later than those with genotype A (58% vs. 37 %).

 

Conclusion:

Children with HBV genotype D showed a significantly higher viral load as

compared to patients with HBV genotype A. Since children with vertical transmission and high viral replication respond significantly poorer to treatment, individuals with genotype D have to be monitored particularly careful. Long-term studies need to clarify, whether these patients are prone to develop a more unfavorable outcome.

 


Abstract ID: 65285

Category: JO9: Hepatitis B: Pathogenesis

Clearance of HBsAg in a cohort of Alaska Natives with Chronic Hepatitis B.

J. Simonetti, Liver Disease and Hepatitis Program, Anchorage, AK, L. Bulkow, Arctic

Investigations Program, Anchorage, AK, C. Homan, Liver Disease and Hepatitis

Program, Anchorage, AK, M. Snowball, Liver Disease and Hepatitis Program,

Anchorage, AK, S. Livingston, Liver Disease and Hepatitis Program, Anchorage, AK,

K. Hurlburt, Liver Disease and Hepatitis Program, Anchorage, AK, J. Williams, Liver

Disease and Hepatitis Program, Anchorage, AK, B. McMahon, Liver Disease and

Hepatitis Program, Anchorage, AK

Disclosures: The following authors have indicated they have no relationships to disclose:

 

Introduction:

Worldwide eight (A-H) Hepatitis B (HBV) genotypes have been identified. The precise role that genotypes play in the natural history of chronic Hepatitis B (HBV) remains to be elucidated. In this study we report the relationship between genotype and clearance of HBsAg in an area where 6 HBV genotypes have been found.

 

Methods:

A long-term study of 1536 HBsAg positive Alaska Natives identified 182

persons who subsequently cleared HBsAg. All chronic HBV carriers were tested biannually for HBsAg. Individuals were considered to have cleared HBsAg when 2 successive specimens tested negative. Genotype, by direct sequencing of the S-gene was determined for 137 of the HBsAg negative individuals for whom stored serum was available. In addition, one HBV DNA level was determined by real-time PCR for 95 individuals an average of 14.5 years after HBsAg clearance (lower limit of detection 25 IU/ml). We analyzed the resultant data to determine if clearance of HBsAg was associated with genotype, age or gender and whether DNA was detectable after HBsAg clearance.

 

Results:

Six genotypes (A, B, C, D, F and H) were identified in this cohort of Alaska

Natives with chronic HBV. The percentage of each genotype that cleared HBsAg was as follows: genotype A 10.6% (13/123), genotype B 4.7% (2/43), genotype C 11.9% (8/67), genotype D 14.7% (85/577), genotype F 13.6% (28/206) and genotype H (1/1). The single genotype H specimen was omitted from further analysis. None of the HBV genotypes found in Alaska were significantly associated with clearance of HBsAg (p = 0.318) even after adjustment for age (p=0.228) and gender (p=0.417). When analyzed by age, there was significantly more HBsAg clearance among persons ≥ 20 years at the time the first HBsAg positive specimen was found (p < 0.01).

Gender was not associated with HBsAg clearance (p=0.688). HBV DNA was found in 15.7% (15/95) of the specimens tested after clearance of HBsAg (range 28-1313 IU/ml) but was not associated with any particular genotype (p=0.469). HCC developed in 3 individuals after they had cleared HBsAg; two were genotype F and one was genotype A.

 

Conclusion:

Clearance of HBsAg was not associated with any of the six genotypes found

in a cohort of Alaska Natives with chronic HBV, nor with gender. Further investigations are required to determine if the viral persistence in those individuals with detectable HBV DNA after clearance of HBsAg is due to viral mutations. These results provide new information about the relationship between HBsAg clearance and genotype. In addition, the development of HCC after HBsAg clearance emphasizes the importance of sustained monitoring after HBsAg seroconversion.

 

 


Poster 1322

Abstract ID: 67571

Category: JO9: Hepatitis B: Pathogenesis

Role of liver biopsy in patients with normal ALT and high HBV DNA.

M. Lai, BIDMC, Boston, MA, B. Hyatt, BIDMC, Boston, MA, N. Afdhal, Beth Israel

Deaconess Medical Center, Boston, MA

 

Background:

Disease progression in hepatitis B (HBV) is very variable and the

importance of clinical factors such as ALT and HBV DNA is unclear. Some authorities (Keeffe et al, Clin Gastroenterol 2004) have suggested that patients with increased HBV DNA and normal ALT should have a liver biopsy.

 

Aim:

To determine the histological stage of fibrosis in patients with normal ALT and elevated HBV DNA.

 

Methods:

We conducted a retrospective chart review of all patients with chronic hepatitis B seen in an outpatient academic Liver Center over 5 years. All patients seen in our Liver Center with the ICD-9 code of 070.32 (chronic hepatitis B) were screened. Inclusion criteria were hepatitis B surface antigen positivity, liver biopsy or clinical cirrhosis, documented prebiopsy

viral loads of greater than 10,000 copies/mL, and lack of treatment prior to

biopsy. We allowed for patients who had been treated with interferon. We compared patients with persistently normal ALT with two other groups: ALT 1-1.5 above upper limits of normal and ALT >1.5 times upper limits of normal. We defined persistently normal ALT as 2 normal ALT measurements 6 months apart and prior to biopsy. Variables of interests were age, weight, height, eAg status, HIV, HCV, prior interferon use, duration of disease, mode of transmission, pre-biopsy viral load, pre-biopsy ALT, and biopsy results including Metavir stage, grade, presence of iron and steatosis. We used the statistical program SAS for Windows 9.0 to perform correlation matrices, univariate and multivariate analyses.

 

Results:

Of the 190 patients who met the inclusion criteria for the study, 57 patients met our criteria for normal ALT, 23 patients with ALT between 1 and 1.5 times the upper limits of normal, and 110 patients with ALT greater than 1.5 times the upper limits of normal. Overall we found three predictors of fibrosis: increasing age (> 43 years), higher ALT and higher grade of inflammation on biopsy. When compared to patients with elevated ALT, patients with normal ALT had lower stages and grades. Predictors of

fibrosis in patients with normal ALT were age (>45yrs) and grade. 23% of normal ALT patients had stage 2 or greater. Neither viral load nor e antigen positivity was predictive of stage in all patients or in the subgroup with normal ALT.

 

Conclusions:

There is significant fibrosis in 24% of chronic hepatitis B patients with normal ALT and elevated HBV DNA. Liver biopsy remains important in identification of these treatment candidates.

 

 

 


Poster 1325

Abstract ID: 64929

Category: JO9: Hepatitis B: Pathogenesis

Hepatitis B Virus - Specific Immune Response After Liver Transplantation.

Y. Luo, Faculty of Medicine, HKU, Hong Kong, China, C. Lo, Faculty of Medicine,

Hong Kong, China, K. Cheung, Faculty of Medicine, HKU, Hong Kong, China, G. Lau,

Faculty of Medicine, HKU, Hong Kong, China, S. Fan, Faculty of Medcine, HKU, Hong

Kong, China, J. Wong, Faculty of Medicine, HKU, Hong Kong, China

 

Introduction:

End-stage liver disease secondary to hepatitis B virus (HBV) has become a universally accepted indication for liver transplantation worldwide as a result of the effective prevention and treatment of recurrent hepatitis B with hepatitis B immunoglobulin and lamivudine. Kinetics of HBV-specific immune response after liver transplantation, which may be influenced by viral clearance and immunosuppression, is poorly understood.

 

Methods:

Here, we studied the serial changes of HBV-specific immune response and the immunological correlates of HBV recurrence after liver transplantation. Frequency and function of circulating HBV-specific T cells from 29 patients with HBV-associated liver disease were studied longitudinally before and after transplantation by proliferation and enzyme-linked immunospot assays. In addition, the HBV-specific immune response was analyzed in 10 patients with breakthrough recurrence of hepatitis B surface antigen (HBsAg) and 11 patients 5 years after liver transplantation for chronic hepatitis B.

 

Results:

After liver transplantation, the HBV-specific T cell reactivity dramatically decreased with a slight enhancement of T cell frequency and intensity of response in the first 3 months followed by a persistent decline. The T cell reactivity to mitogen (phytohemagglutinin) and recall antigen (tetanus toxoid), however, gradually restored after transplantation. The patients with breakthrough recurrence of HBsAg showed significant HBV-specific T immune responses against HBV antigens comparable to non-transplant patients, which were detectable more often in patients with higher level of alanine aminotransferase. In addition, 11 of 29 (37.9%) patients actively developed a transient production of antibody against HBsAg after transplantation, which was significantly associated with HBV immune status of liver donors.

 

Conclusions:

In conclusion, these findings suggest that the decreased HBV-specific immune response may be attributed to viral clearance after removal of diseased liver, and provide a rationale for the active immunoprophylaxis with HBV vaccine at the early stage after transplantation. Despite immunosuppressive treatment, the cellular immune mechanisms have a pathogenic role in graft injury associated with HBV recurrence after liver transplantation.


Poster 1326

Abstract ID: 65789

Category: JO9: Hepatitis B: Pathogenesis

Measurement of Intrahepatic HBV DNA in HBsAg-Negative Patients.

Y. Kim, Digestive Disease Center, Soon Chun Hyang University Hospital, Seoul, Korea,

Republic of, J. Jang, Digestive Disease Center, Soon Chun Hyang University Hospital,

Seoul, Korea, Republic of, Y. Kim, Digestive Disease Center, Soon Chun Hyang

University Hospital, Seoul, Korea, Republic of, C. Han, Digestive Disease Center, Seoul,

Korea, Republic of, S. Eun, Digestive Disease Center, Soon Chun Hyang University

Hospital, Seoul, Korea, Republic of, Y. Cheon, Digestive Disease Center, Soon Chun

Hyang University Hospital, Seoul, Korea, Republic of, J. Moon, Digestive Disease

Center, Soon Chun Hyang University Hospital, Seoul, Korea, Republic of, Y. Cho,

Digestive Disease Center, Seoul, Korea, Republic of, C. Shim, Digestive Disease center,

Soon Chun Hyang University Hospital, Seoul, Korea, Republic of, B. Kim, Digestive

Disease Center, Soon Chun Hyang University Hospital, Seoul, Korea, Republic of

Disclosures: The following authors have indicated they have no relationships to disclose:

Yun Soo Kim, Jae Young Jang, Young Seok Kim, Chang Hee Han, Soo Hoon Eun,

Young Koog Cheon, Jong Ho Moon, Young Deok Cho, Chan Sup Shim, Boo Sung Kim

 

Background:

Occult HBV infection is characterized by presence of HBV infection with

undetectable HBsAg. It has potential risk of HBV transmission through blood transfusion or organ transplantation, and is potentially associated with hepatocellular carcinoma. This study was carried out to find out the frequency HBV infection in patients who lack HBsAg and to identify if there is difference of intrahepatic HBV DNA level according to the serological status.

 

Materials and Methods:

56 HBsAg-negative patients were

included(CH(C) 17, metastatic liver cancer 6, alcoholic liver disease 6, NASH 4, autoimmune liver diseases 4, etc.) Paitents were grouped according to their serological status; group A(cAb+,sAb-, n=16), B(cAb+,sAb+, n=26), and C(cAb-, sAb+/- n=14). Percutaneous liver biopsy were performed using ACECUT¢ç Automatic Biopsy System (TSK lab., Japan, 18G, 15 mm), and liver specimens were immediately frozen and

stored at - 70 C until DNA extration. After DNA extration, HBV DNA quantification was performed using RealArt¢â HBV LC PCR Reagent (Artus, Germany) with Lightcycler¢ç Instrument (Roche Diagnostics, Germany).

 

Results:

Overall frequency of detectable intrahepatic HBV DNA was 34%(19/56). The frequency was 56%(9/16) in Gr A, 31%(8/26) in Gr B and 14%(2/14) in Gr C(p =0.01). Levels of intrahepatic HBV DNA were as follows; 201 ¡¾ 666 copies/mg in Gr A, 618 ¡¾ 2953 copies/mg in Gr B and 35 ¡¾ 122 copies/mg in Gr C, which were statistically insignificant. HCV infection was not associated with high prevalence of occult HBV infection.

 

Conclusion:

Low level of intrahepatic HBV DNA is frequently found in anti-HBc positive patients, even though anti-HBs is positive.

 


Poster 1327

Abstract ID: 66569

Category: JO9: Hepatitis B: Pathogenesis

High Prevalence of Occult Hepatitis B in Patients with Advanced Liver Disease and Patients with Hepatitis C.

A. A. Raouf, National Liver Institute, Shebeen EL Kom, Egypt, H. El Saeed, National

Liver Institute, Shebeen El Kom, Egypt, N. Ghnaiem, Faculty of Medicine, Shebeen El

Kom, Egypt, A. Bassiouny, National Liver Institute, Shebeen El Kom, Egypt, H. El

Ghazzawy, National Liver Institute, Shebeen El Kom, Egypt, I. Waked, National Liver

Institute, Cairo, Egypt

 

Introduction:

Several reports indicated the higher prevalence of occult hepatitis B infection in patients with chronic hepatitis C, and that occult HBV might influence the clinical and biochemical features, and the severity of disease. This has not been studied among Egyptian patients with chronic HCV infection due to HCV genotype 4.

 

Aim:

To evaluate the presence of HBV-DNA in HBsAg negative patients with chronic liver disease, and to correlate its presence with HCV infection, and with severity of liver disease.

 

Patients and Methods:

224 HBsAg negative patients with liver disease and 227 HBsAg

negative and HCV negative individuals without liver disease had serum HBV-DNA tested using nested PCR by specific primers of surface antigen. All procedures were performed in duplicate. Patients were: 185 HCV genotype 4 positive (86 with liver cirrhosis, 58 with chronic hepatitis, 41 with persistently normal ALT) and 39 HCV negative (14 with liver cirrhosis and 25 with chronic active hepatitis).

 

Results:

HBV-DNA was positive among 53 of 100 patients with liver cirrhosis (54% in HCV+ve vs 43% in HCV-ve, ns). Of these, 5 were serongative for all HBV markers. Similarly, in CAH, the prevalence of HBV-DNA did not differ among patients with and without HCV infection.

 

HBV was more prevalent among patients with liver disease than controls (p<0.001, OR45, 95%CI: 11.59-382), among patients with HCV infection than controls (P<0.001, OR50.1, 95%CI: 12.8-427)), and among patients with advanced than mild liver disease (cirrhosis vs CAH p<0.005, OR8.78, 95%CI 3.4-25.2). No significant biochemical differences were observed among HBV-DNA positive or negative patients with similar stages of liver disease, and the Child-Pugh score and clinical indications of severity were

similar among HBV-DNA positive and negative patients with cirrhosis.

 

Conclusion:

Occult hepatitis B is highly prevalent among patients with hepatitis C and

with liver disease compared to control, and the prevalence increases with advanced liver disease. Among patients with similar degree of liver disease the presence of HBV-DNA is not associated with increased markers of disease severity. The role of occult hepatitis B in the progression of liver disease remains to be identified in future studies.

 

 


Poster 1328

Abstract ID: 66636

Category: JO9: Hepatitis B: Pathogenesis

Viral factors which predicted liver-related deaths and development of hepatocellular carcinoma in patients with chronic hepatitis B.

M. J. Tong, Liver Center, Huntington Medical Research Institutes, Pasadena, CA, L. M.

Blatt, Intermune, Inc., Brisbane, CA, J. Kao, Hepatitis Research Center, National Taiwan

University Hospital, Taipei, Taiwan, J. T. Cheng, Liver Center, Huntington Medical

Research Institutes, Pasadena, CA, W. G. Corey, Liver Center, Huntington Medical

Research Institutes, Pasadena, CA

 

Background:

Chronic hepatitis B patients are at risk for death from complications of cirrhosis and for progression to Hepatocellular carcinoma (HCC). To identify risk factors which are associated with these complications, we conducted a prospective study in HBsAg positive patients followed in our Liver Center.

 

Patients and Methods:

From January 1989 to March 1998, we enrolled 440 patients who were followed for a mean of 117±58 SD months. Serum samples were obtained during the initial visit, stored at −70°C, and then sent to the Hepatitis Research Center at National Taiwan University Hospital in Taipei, Taiwan in 2004. Sera were tested for HBV-DNA, HBV genotype, precore (PC) mutation and basal core promoter (BCP) mutation. Also, baseline liver tests, platelet counts, HBeAg and anti-HBe were obtained. Patients were divided into inactive carriers (IC), mild chronic hepatitis (MCH), chronic active hepatitis (CAH), cirrhosis and HCC. Thereafter, patients without HCC were followed for death from cirrhosis or development of HCC.

 

Results:

At baseline, 49% of patients were HBeAg positive and 51% were anti-HBe

positive. HBV genotype B was more common in IC, and genotype C was predominant in patients with CAH, cirrhosis and HCC (P=0.004). Precore mutation was more often detected in genotype B (P<0.0001) and in HCC patients (P=0.0005), while BCP mutation was more common in genotype C patients (P<0.0001) and in HCC patients (P=0.005). Inactive carriers and HCC patients had the lowest mean HBV-DNA levels than patients in other categories (P<0.0001). Also, genotype C patients had the highest mean HBVDNA titers (P<0.009). Compared to their wild-type counterparts, PC mutants and BCP mutants had lower mean HBV-DNA values (P<0.0007 for both). During follow-up, 30 patients developed HCC and 38 patients died from non-HCC related liver deaths. Stepwise logistic regression analysis identified male sex, baseline bilirubin and platelet values and higher HBV-DNA levels as independent predictors of non-HCC related liver deaths, while age, PC mutation and BCP mutation were independent predictors for HCC development.

 

Conclusions:

Hepatitis B PC mutation and BCP mutation were highly associated with

development of HCC. On the other hand, tests indicating … of liver disease and high circulating levels of HBV-DNA identified patients who died from non-HCC related liver deaths.

 


Poster 1329

Abstract ID: 66978

Category: JO9: Hepatitis B: Pathogenesis

Time-Dependent Relative Risk Of Hepatocellular Carcinoma (HCC) For Markers Of Chronic Hepatitis B (CHB): The R.E.V.E.A.L.-HBV Study.

C. Chen, National Taiwan University, Taipei, Taiwan, H. Yang, National Taiwan

University, Taipei, Taiwan, U. H. Iloeje, Bristol-Myers Squibb Pharmaceutical Research

Institute, Wallingford, CT, J. Su, Bristol-Myers Squibb Pharmaceutical Research

Institute, Wallingford, CT, C. Jen, National Taiwan University, Taipei, Taiwan, S. You,

National Taiwan University, Taipei, Taiwan, Y. Liaw, Chang Gung Memorial Hospital

and University, Taipei , Taiwan

 

Introduction:

Progression of CHB infection to HCC is a long-term process. The impact

of predictors may change over time. Studies have estimated relative risk of HCC predictors assuming they had fixed effect on hepatocarcinogenesis. We assessed relative risk associated with different risk factors for HCC at different times over a long period of follow-up.

 

Methods:

A cohort of 3,653 HBsAg-seropositive and anti-HCV-seronegative

residents enrolled from 7 townships in Taiwan between 1991 and 1992 was followed for newly occurring HCC through June 30, 2004. HBsAg and HBeAg status, serum alanine aminotransferase (ALT), HBV DNA levels, and liver cirrhosis (detected by abdominal ultrasonography) were evaluated at cohort entry. Newly developed HCC was ascertained through follow-up health examinations by serological tests and abdominal ultrasonography as well as by computerized data linkage to the national cancer registry and death certification system. Cox’s proportional hazards regression analyses with stratification of HCC diagnosis date into three strata (<5, >5 -9 and >9 years after enrollment) were used to assess the changing relative risk (RR) with its 95% confidence interval (CI) for various risk predictors after adjustment for age and gender.

 

Results:

In the follow-up of 41,779 person-years, 58, 50, 56 new HCC cases occurred during the three consecutive periods of <5, >5-9 and >9 years after enrollment respectively. The pattern of predictive risk factors varied for HCC occurring during each period of followup time. The age-gender-adjusted RRs (95% CI) for habitual alcohol consumption was significant only for HCC occurring after 9 years; 1.6 (0.9-3.1), 1.3 (0.6-2.6) and 3.0 (1.6- 5.5), but increased over time for HBeAg-seropositivity; 5.6 (3.2-9.7), 7.0 (3.9-12.5) and 9.0 (5.2-15.6). The risk associated with elevated ALT (>45 IU) was steady for HCC occurring over time; 3.8 (2.0-7.3), 5.4 (2.8-10.4), 3.2 (1.5-6.8). There were decreasing RRs of 20.3 (11.4-36.1), 15.0 (7.4-30.5), and 6.4 (2.3-17.9) for liver cirrhosis detected by ultrasonography. A significant dose-response relationship was observed between elevated

serum HBV DNA level and risk of newly developed HCC. For HBV DNA >106 copies/mL, the age- and gender-adjusted RRs for HCC increased over time from 7.7 (3.1- 19.1), and 17.4 (5.2-57.9), to 35.0 (8.3-147.7).

 

Conclusion:

These data suggest that liver cirrhosis is associated with increased near term risk of HCC. Elevated ALT level conferred a steady risk over time. Habitual alcohol consumption, HBeAg-seropositivity and elevated HBV DNA level strongly predict risk over time, and the risk associated with elevated HBV DNA increases over time.
Abstract ID: 67077

Category: JO9: Hepatitis B: Pathogenesis

Prevalence and impact of occult HBV infection on severity and response to therapy of chronic hepatitis C in France.

S. Mrani, INSERM, LYON, France, I. Chemin, INSERM U271, LYON, France, K.

Menouar, INSERM U271, LYON, France, O. Guillaud, INSERM U271, LYON, France,

G. Borghi, Liver Unit, Hôtel Dieu Hospital, LYON, France, P. Pradat, Liver Unit, Hôtel

Dieu Hospital, LYON, France, M. Trabaud, Laboratoire de virologie, Hôpital de la

Croix-Rousse, LYON, France, P. Chevallier, Laboratoire de virologie, Hôpital de la

Croix-Rousse, LYON, France, F. Zoulim, INSERM U271, LYON, France, C. Trépo,

INSERM U271, LYON, France

 

Introduction:

Frequent co-infection of occult HBV infection in chronic hepatitis C patients has been reported. However, its prevalence and its impact on outcome are still a matter of controversy.

 

Aim:

The aim of this study was to establish the frequency of this phenomenon among French HCV chronic carriers and secondly to better understand the role of “occult” HBV infection during the natural history of HCV and its implications. A cohort of 203 patients with chronic hepatitis C and without HBsAg has been examined. Serum HBV DNA was detected using a highly sensitive PCR with two sets of primers located in the S and X genes respectively. Furthermore, HBV viremia levels were evaluated by a real time PCR in the C gene of HBV.

 

Results:

The results showed that 47 patients (23, 15%) had occult HBV infection all with a low HBV titer (102-104 copies/ml). No significant difference in age, gender, serum ALT level, HCV genotypes and the presence of anti-HBc was observed in patients with or without HBV DNA. However, the histological lesions were more severe in HBV DNA positive cases than among negative ones both regarding Metavir activity (53 vs 38%, P<0.01) and fibrosis scores (60 vs 33%, P<0.001). In addition, a sustained virological response to classical IFN/Ribavirin combination therapy against chronic hepatitis C was

achieved in 11 (28%) of 40 HBV DNA positive cases, compared with 65 (45%) of the 144 HBV DNA negative cases (P<0.05).

 

Conclusion:

In conclusion, HBV DNA is found in ¼ of French HCV chronic carriers without HBsAg irrespective of the presence of anti-HBc. Preliminary findings do suggest that occult HBV infection increases the severity of liver disease and favors the resistance to antiviral therapy in chronic hepatitis C.