Hepatitis B Treatment  11/14/2005 8:00 AM - 6:30 PM

 

 

Poster 960

Abstract ID: 66805

Category: J1O: Hepatitis B: Treatment

One-year treatment of entecavir results in reduction in intrahepatic covalently closed circular DNA level.

D. K. Wong, The University of Hong Kong, Hong Kong, Hong Kong, M. Yuen, The University of Hong Kong, Hong Kong, Hong Kong, V. Ngai, The University of Hong Kong, Hong Kong, Hong Kong, C. Kwok, The University of Hong Kong, Hong Kong, Hong Kong, C. Lai, The University of Hong Kong, Hong Kong, Hong Kong

 

Background:

Two phase 3, multicenter, double-blind trials on HBeAg-positive and HBeAg-negative patients demonstrate that, compared with lamivudine, entecavir is superior in inducing histologic improvement, serum HBV DNA suppression and transaminase normalization. Since intrahepatic HBV DNA and covalently closed circular (ccc) DNA are important for control of viral replication, the efficacy of entecavir vs. lamivudine in achieving suppression of these entities should be studied.

 

Methods and Patients:

The patients involved in this study were recruited from patients

participating in the two phase 3 entecavir trials at our center in Hong Kong. Forty chronic hepatitis B patients (14 HBeAg-positive and 26 anti-HBe-positive) were randomized to receive either entecavir (0.5 mg once daily) or lamivudine (100 mg once daily). Paired liver biopsy and serum samples were collected both at baseline and week 48 of treatment. Total intrahepatic HBV DNA and cccDNA were measured by the Invader® assay.i Serum HBV DNA was measured by the COBAS Amplicor HBV Monitor Test.

 

Results:

This is an interim report for the 14 HBeAg-positive patients. (The findings of the 26 anti-HBe-positive patients will be analyzed later.) For the 14 HBeAg-positive patients, 7 were randomized to receive entecavir, while the rest received lamivudine.

 

 

Conclusion:

The preliminary results of this study showed that 1 year of entecavir was

superior to lamivudine in suppression of total intrahepatic and ccc DNA in liver biopsies. The addition of the data from the 26 anti-HBe patients may confirm these results.

 

i Wong et al, Hepatology (2004) 40:727-737.

 


Poster 961

Abstract ID: 61589

Category: J1O: Hepatitis B: Treatment

 

High prevalence of significant fibrosis in patients with immunotolerance to chronichepatitis B infection.

C. Wang, University of Washington, Seattle, WA, H. Deubner, University of Washington, Seattle, WA, M. Shuhart, University of Washington, Seattle, WA, J. N. Manansala, University of Washington, Seattle, WA, L. Corey, University of Washington, Seattle, WA, K. V. Kowdley, University of Washington, Seattle, WA

 

OBJECTIVE:

To determine the extent of liver disease in patients with chronic hepatitis

B and normal serum ALT.

 

METHODS:

Patients were eligible with HBeAg positive CHB, serum HBV DNA > 106

copies/mL, and 2 ALT measurements within normal limits in the 2 years prior to liver biopsy. The mean value of the 2 ALT measurements was calculated. Liver biopsies were scored using the Batts and Ludwig scoring method with a 4-point scoring scale for inflammation and fibrosis.

 

RESULTS:

9 men and 4 women were enrolled. Median age was 26 (range 19-46). All

had endemically-acquired infection by history. Nine patients were born in China, 2 in Vietnam, 1 in Korea, and 1 in Cambodia. Median ALT was 28 U/L (range 13-77) and median serum HBV DNA was 5.1 x 107 (range 4.5 x 104 – 3.4 x108). On biopsy, 10 of 13 (77%) had evidence of fibrosis: None had cirrhosis or septal fibrosis; 6 of 13 (46%) had periportal fibrosis or rare portal-portal septa, resulting in fibrosis scores of 2, and 4 of 13 (31%) patients had portal fibrosis with at most mild periportal fibrosis, yielding fibrosis scores of 1 or 1-2. Mean liver biopsy grade was 1.23 (range 1-2) and mean stage was 1.30 (range 0-2). The R-square coefficient for a correlation between ALT and liver biopsy stage was .07, indicating that ALT was poorly correlated with liver biopsy fibrosis score (see graph). Four of 5 patients with a liver fibrosis stage of 2 had serum ALT < 30 IU/L.

 

CONCLUSIONS:

A high proportion of Asian immunotolerant HBeAg positive patients

have hepatic fibrosis on biopsy. Serum ALT level may not accurately predict histologic stage in these patients. These preliminary findings support recent recommendations to perform liver biopsy to guide treatment decisions in HBeAg positive patients even if serum ALT is normal. Supported by a research grant from Gilead Sciences (Wang) and by K24 DK02957 (Kowdley).

 


Poster 962

Abstract ID: 67445

Category: J1O: Hepatitis B: Treatment

Entecavir Two Year Resistance Update: No Resistance Observed in Nucleoside Naïve Patients and Low Frequency Resistance Emergence in Lamivudine Refractory Patients.

 

R. Colonno, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, R. Rose, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, S. Levine, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, J. Baldick, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, K. Pokornowski, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, M. Plym, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, C. Yu, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, C. Mazzucco, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, J. Fang, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, M. Hsu, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, A. Walsh, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, B. Eggers, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CO, A. Thiry, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, D. Tenney, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT

 

Background:

Entecavir (ETV) is a potent inhibitor of hepatitis B virus (HBV) with proven

clinical efficacy. High level ETV resistance (ETVr) requires pre-existing lamivudine (LVD)- resistance (LVDr) substitutions and additional changes at HBV RT residues rtT184, rtS202 or rtM250. Patients receiving ETV for 1 year showed no evidence of emerging ETVr in nucleoside naïve patients and virologic rebounds due to ETVr in only 1% of LVD refractory patients.

 

Methods:

Virologic rebounds (confirmed ³1 log increase from nadir by PCR) observed in patients from studies AI463-022 & AI463-027 (nucleoside naïve), AI463-014 & AI463-026 (LVD refractory), and AI463-901 (extended treatment) were analyzed for emerging ETVr. Genotypic analysis compared patient HBV RT sequences with those at study entry and with WT HBV. Phenotypes of emerging substitutions were determined using antiviral assays measuring HBV DNA yields from HepG2 cells transfected with plasmids expressing patient RT sequences.

 

Results:

Among >650 nucleoside naïve, HBeAg pos & HBeAg neg patients completing at least 24 wk of ETV therapy, 93% achieved HBV DNA reductions to <300 copies/ml. Over 200 patients completed ≥ 90 wks of therapy. Genotypic analysis of the 18 observed virologic rebounds (2 year treatment period) failed to show any evidence of emerging ETVr substitutions, with population phenotypes at the time of rebound essentially unchanged from baseline or WT. Four additional patients failing to achieve HBV DNA reductions <100,000 copies/ml on ETV also had HBV fully susceptible to ETV. Therefore, there is no evidence of ETVr emerging in nucleoside naïve patients treated with ETV. Virologic rebounds due to resistance were observed in 10% of LVD refractory patients treated with ETV for 2 years. In all cases, ETVr variants had pre-existing LVDr substitutions and emerging changes at residues rtT184 and/or rtS202. Of the 12 patients exhibiting a rebound, 2 had been on suboptimal ETV therapy (0.5 mg) and 3 others received ETV/LVD combination therapy. Subsequent analysis of baseline viral samples demonstrated selection of ETVr substitutions during prior LVD treatment.

 

Summary:

There was no evidence of emerging ETVr in nucleoside treatment naïve subjects undergoing 2 years of ETV therapy, coinciding with substantial suppression of viral DNA levels. Among LVD refractory patients,

10% experienced virologic rebounds due to emerging ETVr by the 2nd yr of therapy. Phenotypic ETVr required the presence of pre-existing LVDr substitutions, which can be selected with exposure to LVD. Therefore, LVD treatment results in frequent emergence of LVDr and may negatively impact future HBV treatment options.

 

 


Poster 963

Abstract ID: 61542

Category: J1O: Hepatitis B: Treatment

 

REAPPEARANCE OF WILD-TYPE HEPATITIS B VIRUS DURING ADEFOVIR MONOTHERAPY FOR PATIENTS WITH LAMIVUDINE RESISTANCE.

 

H. Hann, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA, J. L. Platt, Questdiagnostics Nichols Institute, San Juan Capistrano, CA Disclosures: Hie-Won Hann - Speakers Bureau(s):GlaxoSmtihkline Gilead Sciences,Inc., Consultant for: GlaxoSmtihkline Gilead Sciences,Inc., Grant / Research Support by: GlaxoSmtihkline Gilead Sciences,Inc.; Jamie Platt

 

Background?:

No relationships to disclose Adefovir Dipivoxil (ADV) is effective for both wild type (WT) and YMDD variant (YMDDv) HBV which is resistant to Lamivudine (LAM). While the majority of LAM resistant patients respond well to ADV, some patients show slow/poor antiviral responses and even have an increase in HBV DNA levels while on ADV therapy. We investigated if this poor/worsening response was the result of ADV resistance.

 

Materials and Methods:

133 patients after LAM resistance, were treated with ADV for varying periods. Good responses to ADV were seen in 87/133 patients while 46 showed slow /poor virologic responses although biochemical improvement was noted. Pre-ADV and on-ADV therapy serum samples were available from 13/46 patients. For the remaining 33, Pre-ADV and on-ADV specimens were unavailable since HBV DNA levels were measured elsewhere. To understand the nature of slow/poor responses, HBV polymerase genotypes were investigated using an YMDD PCR-RFLP assay and sequencing of the HBV pol gene. The PCR-RFLP assay examined the presence of mutations at 2 sites (rtL180 and rtM204)). The sequencing assay assessed mutations at codons, rt180, rt181, rt204, and rt236. Analytes of the sequencing assay included genotype and PMUL and PMUA, polymerase mutants for LAM and for ADV espectively. These 13 patients were Asian Americans, aged 19-67 years, 9 males and 4 females. Treatment with LAM lasted 12-60 months until viral breakthroughs were noted. Median pre-ADV HBV DNA levels were 5 x 106 copies/mL (range 1.2 x 106-2.5 x 108) while median on-ADV HBV DNA levels were 9 x 106 (range 1.7x 104 - 8 x 106). The median duration of ADV therapy for 13 patients was 13 months (range 6-30).

 

Results:

Polymerase genotypes were examined on 10 pre-ADV and 13 on-ADV samples, all of HBV genotype C. Among 10 pre-ADV samples, 8 contained YMDDv HBV and 5/8 also contained a small portion of mixed population; surprisingly 2/10 had mostly WT-HBV although they had viral breakthrough. Following ADV therapy (6-30 months), 10/13 patients had a return of WT-HBV; 3 of these10 contained a small portion of YMDD and 3/13 had YMDDv-HBV. One patient with 18months on ADV, showed WTHBV return mixed with ADV resistant HBV (N236T). It appeared that YMDDv-HBV were suppressed by ADV but in some WT-HBV was strong enough to return even under ADV effect.

 

Conclusion:

These results suggest that in patients with LAM resistance who did not

respond well to ADV, the majority of poor responses were due to the return of WT HBV and not due to ADV resistance. Continuation of LAM in combination with ADV, at least for a period of time, may therefore be the preferred management in patients with LAM

resistance.

 


Poster 964

Abstract ID: 63945

Category: J1O: Hepatitis B: Treatment

 

First line combination therapy of chronic hepatitis B with tenofovir plus lamivudine versus sequential therapy with tenofovir monotherapy after lamivudine failure.

 

S. Mauss, Center for HIV and Hepatogastroenterology, Duesseldorf, AE, Germany, M. Nelson, Kobler Clinic, Chelsea, AE, United Kingdom (Great Britain), T. Lutz, HIVPractice,Frankfurt, AE, Germany, J. Sheldon, Hospital Carlos III, Madrid, AE, Spain, R. Bruno, San Matteo Hospital, Pavia, AE, Italy, F. van Boemmel, Charite, Berlin, AE, Germany, J. Rockstroh, Med. Klinik I, Bonn, AE, Germany, E. Wolf, MUC Research, Muenchen, AE, Germany, A. Stoehr, Infektiologie, Hamburg, AE, Germany, V. Soriano, Hospital Carlos III, Madrid, AE, Spain, F. Berger, Center for HIV and Hepatogastroenterology, Duesseldorf, AE, Germany, T. Berg, Charite, Berlin, AE, Germany, A. Carlebach, HIV-practice, Frankfurt, AE, Germany, C. Schwarze-Zander, Med. Klinik I, Bonn, AE, Germany, T. Wunsche, Charite, Berlin, AE, Germany, H. Jaeger, MUC Research, Muenchen, AE, Germany, G. Schmutz, Center for HIV and Hepatogastroenterology, Duesseldorf, AE, Germany

 

Therapy with HBV-polymerase inhibitors is based on long term suppression of viral replication. At present sequential monotherapy is standard of care, however combination therapy with at least two HBV-polymerase inhibitors is considered a promising alternative approach. It is unknown to date whether combination therapy with lamivudine plus tenofovir could be superior to sequential therapy with tenofovir after the occurrence of lamivudine resistance for chronic hepatitis B treatment in coinfected patients.

 

We conducted a multicenter, 1:2 matched pair analysis comparing patients with HBV/HIV-coinfection starting an antiretroviral regimen including tenofovir plus lamivudine with patients who had highly replicative, lamivudine resistant hepatitis B (> 100,000 copies/mL) and started therapy with tenofovir as the only active HBV-polymerase inhibitor. Resistance to lamivudine was demonstrated by HBVgenotyping. The lower limit of detection of HBV-DNA was 1000 copies/mL. At baseline patients starting with tenofovir plus lamivudine (n=23) had a median HBV-DNA of 59,000,000 copies/mL compared to 120,000,000 copies/mL in the tenofovir arm (n=46) (p=0.75). After 3 months on treatment, median HBV-DNA decreased to 138,450 copies/mL in the tenofovir plus lamivudine group compared to 27,950 copies/mL in the tenofovir group (p=0.26). After 12 months and 24 months median HBV-DNA was <1000 copies/mL in patients taking either tenofovir plus lamivudine or tenofovir alone (p=0.46, p=0.24). A sustained undetectable HBV-DNA <1000 copies/mL was achieved in 19/23 (83%) patients on tenofovir plus lamivudine and in 38/46 (83%) patients on tenofovir (p = 1.00). A loss of HBe-antigen was observed in 7/22 HBe-antigen positive patients on tenofovir plus lamivudine and in 11/44 patients on tenofovir (p=0.57). HBsantigen loss was found in 1/23 and 2/46 patients, respectively.

 

In conclusion in this cohort of HBV/HIV-coinfected individuals, full virologic HBVDNA suppression was achieved in the majority of patients independent of treatment allocation. In addition loss of HBe- and HBs-antigen was not different in both arms. Over a median observational period of 24 months tenofovir alone was as effective as tenofovir plus lamivudine in HBV/HIV-coinfected patients with highly replicative chronic hepatitis B.


Poster 965

Abstract ID: 64736

Category: J1O: Hepatitis B: Treatment

 

Hepatitis B Virus with rtL80V/I Mutation Associates with Poor Response to Adefovir Dipivoxil Therapy.

Y. Lee, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea,Republic of, Y. Chung, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea, Republic of, S. Ryu, University of Inje College of Medicine, Seoul Paik Hospital, Seoul, Korea, Republic of, J. A. Kim, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea, Republic of, M. Choi, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea, Republic of, S. Jung, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea, Republic of, S. Kim, Korea Veterans' Hospital, Seoul, Korea, Republic of, J. Shin, University of Ulsan College of Medicine, Ulsan University Hospital, Ulsan, Korea, Republic of, N. Park, University of Ulsan College of Medicine, Ulsan University Hospital, Ulsan, Korea, Republic of, K. Kim, University of Ulsan College of Medicine, Seoul, Korea, Republic of,  Y. Lim, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea, Republic of, Y. Lee, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea, Republic of, D. Suh, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea, Republic of

 

Background/Aims:

Resistance occurs frequently during long-term lamivudine (LAM)

therapy, mostly associated with YMDD mutants (rtM204V/I). Besides rtM204V/I mutation, rtL80V/I and rtL180M mutations have been reported to be associated commonly in patients with LAM resistance. In this study, we intended to examine the effects of a certain type of mutations at polymerase domain of Hepatitis B Virus (HBV) on the antiviral efficacy following adefovir dipivoxil (ADV) therapy in patients with LAM-resistant chronic hepatitis B (CHB).

 

Methods:

One hundred and nineteen patients with LAM-resistant CHB were treated with ADV at a dose of 10mg daily for >12 weeks. The entire polymerase domain of HBV in the sera obtained just before ADV therapy was sequenced using direct sequencing method. Genotypes of HBV were determined by restriction fragment length polymorphism (RFLP) following polymerase chain reaction (PCR). Serum HBV-DNA levels were quantified by real-time PCR method. The antiviral responses, which were evaluated by the change of serum alanine aminotransferase (ALT) and HBV-DNA levels at 12 weeks following ADV therapy, were compared in relation to the preexisting mutations at HBV polymerase domain.

 

Results:

All of 119 subjects revealed to have HBV of genotype C. Out of them, 105 patients (88%) had YMDD mutations; 70 (67%) rtM204I, 28 (27%) rtM204V variant and 7 (6%) both. In addition to mutations at YMDD motif, rtL80V/I mutation was found in 70 (59%) and rtL180M in 72 (61%). The rtM204I variant was accompanied by rtL80V/I (p<0.001) and rtM204V was associated with rtL180M mutation more frequently (p<0.005). Median serum HBV DNA level at baseline was 7.82 log(10) copies/mL and the median change of serum HBV DNA levels from baseline were -2.40 log(10) copies/mL at week 12. The rate of serum ALT normalization was 42% following 12 weeks of ADV therapy. The rate of serum ALT normalization (45% vs. 35%; p>0.05) and the changes of serum HBV DNA level (median, -2.43 vs. -2.93 log(10) copies/mL; p>0.05) at 12 weeks following ADV therapy were not significantly different between patients with rtM204I and those with rtM204V variant. Very interestingly, the change of serum HBV-DNA levels was significantly greater in patients with rtL80V/I mutation compared with those without it. (Median, -2.10 vs. -2.83 log(10) copies/mL; p<0.05) However, the presence of rtL180M mutation did not affect the change of serum HBV-DNA level following ADV therapy. (Median, -2.53 vs. -2.20 log(10) copies/mL; p>0.05)

 

Conclusions:

It is suggested that rtL80V/I variants of HBV may associate with poor antiviral response to ADV therapy.


Poster 966

Abstract ID: 67006

Category: J1O: Hepatitis B: Treatment

 

Delayed viral decline is associated with highest sustained response rate during PEGinterferon treatment for HBeAg-positive chronic hepatitis B.

M. J. ter Borg, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands, H. J. Flink, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands, H. L. Janssen, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands, B. E. Hansen, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands, R. A. de Man, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands, S. W. Schalm, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands

 

Little is known about the patterns of HBV-DNA decline and their relation to response for chronic hepatitis B patients treated with alpha-interferon. To investigate different patterns in viral decline during treatment and follow-up, we analyzed 254 HBeAg-positive chronic hepatitis B patients treated with pegylated interferon alpha-2b (PEG-IFN) 100 μg/week for 52 weeks with or without lamivudine 100 mg/day. PEG-IFN dose was reduced to 50 μg/week after 32 weeks of treatment. Endpoints were HBeAg-negativity, HBV-DNA < 400 copies/ml and HBsAg negativity 26 weeks after therapy.

 

The total trial population consisted of 266 patients. From 12 patients, insufficient HBV-DNA measurements were available to assess HBV-DNA patterns.

 

In the patients treated with PEG-IFN monotherapy (n=124), 5 different patterns of viral decline could be recognized: a. early decline of at least 1 log during week 0-4 of therapy; b. delayed decline of at least 2 log from baseline HBV-DNA during week 4-32; c. late decline of at least 2 log between week 32 and 52; d. post-treatment decline of 2 log from baseline HBV-DNA after week 52; e. no substantial decline at any time point. Endpoints for these different patterns are shown in the table. A delayed rather than early viral decline was associated with highest response rates at the end of follow-up (HBeAg response 63% vs. 52%, respectively).

 

Patients with a late or post treatment decline pattern had lower response rates (HBeAg response 31% and 27%, respectively).

 

Interestingly, 7 out of 8 patients (88%) with HBsAg loss and all patients with HBV-DNA <400 copies/ml at the end of follow-up exhibited a delayed HBV-DNA decline. In the patients treated with combination therapy (n=130), a similar biphasic pattern of decline in HBV-DNA was seen in nearly all patients with a fast decline in the first four weeks and a slower decline thereafter.

 

In conclusion, different patterns in HBV-DNA decline were found during PEG-IFN monotherapy. A delayed rather than early viral decline was associated with the highest response rate. This underlines the important immunomodulatory effect of PEG-IFN and the limited predictive value of early viral kinetics in PEG-IFN therapy for HBeAg-positive chronic hepatitis B.

 

PEG Monotherapy


Poster 967

Abstract ID: 67180

Category: J1O: Hepatitis B: Treatment

 

Effect of ethnicity, genotype, gender, age and bodyweight on sustained response in a large, randomised study of peginterferon alfa-2a (40KD) (PEGASYS®) +/- lamivudine versus lamivudine alone for HBeAg-positive chronic hepatitis B.

 

W. Chow, Singapore General Hospital, Singapore, Singapore, M. Manns, Medizinischen Hochschule, Hannover, Germany, S. Paik, Samsung Medical Center, Seoul, Korea, Republic of, T. Berg, Charité Universitätsmedizin Berlin, Berlin, Germany, T. Piratvisuth, Songklanakarin Hospital, Songkla, Thailand, W. Chang, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, G. K. Lau, Queen Mary Hospital, Hong Kong, China, P. Marcellin, Hôpital Beaujon, Clichy, France, E. Gane, Middlemore Hospital, Otahuhu, New Zealand, N. Pluck, Roche, Welwyn, United Kingdom (Great Britain)

 

Background:

Recent data have shown that baseline ALT, HBV DNA and HBeAg levels are strongly associated with sustained HBeAg seroconversion in patients treated with peginterferon alfa-2a and/or lamivudine for chronic hepatitis B (CHB).

 

Objective:

To investigate the effect of ethnicity, genotype, gender, age and bodyweight on response in patients receiving peginterferon alfa-2a ± lamivudine or lamivudine alone for CHB.

 

Methods:

HBeAg-positive patients (n=814) received 48 weeks of 180mg peginterferon alfa-2a (PEGASYS®) once-weekly (qw) + placebo daily (qd), 180mg peginterferon alfa-2a qw + 100mg lamivudine qd, or 100mg lamivudine qd. For this analysis, response was defined as HBeAg seroconversion 24 weeks post-treatment (week 72). Baseline variables included in multivariate (MV) analysis: ethnicity, genotype, gender, age, bodyweight, ALT, HBV DNA and HBeAg.

 

Results:

Most patients were of Asian origin (87%) and infected with HBV genotype C (59%) or B (28%). Response in Asian patients closely reflected overall results. Among Caucasians (n=79), the highest response rates were seen with peginterferon alfa-2a alone (50% [12/24]). Response rates were also high in patients with genotype A receiving peginterferon alfa-2a alone (52% [12/23]). Patients infected with genotype C or B had the same response to peginterferon alfa-2a alone (31% vs 30%). Response rates were higher in females than males in all treatment arms; 35% vs 31%, 37% vs 25%, and 26% vs 17% with peginterferon alfa-2a alone, combination therapy, and lamivudine alone. There was no clear relationship between patient age and response, regardless of treatment. In the two peginterferon alfa-2a arms, response rates were comparable in patients weighing ≤ 65 kg and >65 kg. Response rates in lamivudine-treated patients weighing ≤ 65 kg were slightly higher than in those >65 kg (22% vs16%). In MV analyses across all treatment arms, ethnicity, genotype, age and bodyweight were not significant predictors of response (P=0.757, ≥ 0.21, 0.716 and 0.790); gender was of borderline significance (P=0.077).

 

Conclusion:

Genotype was not a significant predictor of response by MV analysis. The rate of response was the same for patients with genotype B or C, the predominant genotypes in the study. This contrasts with previous studies of interferon-based therapy. Genotype A patients, who represented a minority of patients in our study, treated with peginterferon alfa-2a alone had the

highest rate of HBeAg seroconversion at week 72. Gender had a marginal effect on response and this was mostly seen with combination therapy and lamivudine alone. Patient bodyweight had no effect on response to peginterferon alfa-2a.


Poster 969

Abstract ID: 62802

Category: J1O: Hepatitis B: Treatment

 

A sensitive line probe assay to simultaneously detect Lamivudine and Adefovir resistant mutations.

 

M. T. Hussain, University of Michigan, Ann Arbor, MI, S. Fung, University of Michigan, Ann Arbor, MI, J. Doutreloigne, Innogenetics, Inc, Ghent, Belgium, E. Sablon, Innogenetics, Inc, Ghent, Belgium, A. S. Lok, University of Michigan, Ann Arbor, MI

 

Background:

Antiviral-resistant hepatitis B virus (HBV) mutations have become an increasing problem in the treatment of chronic hepatitis B. Development of rapid, simple, and sensitive assays that can detect antiviral-resistant HBV as they emerge may help improve patient management.

 

Aim:

To assess the accuracy of a line probe assay, INNOLiPA HBV DRv2 (Innogenetics NV, Ghent, Belgium), by comparing results of this assay with those of direct sequencing and to determine if DRv2 can detect antiviral-resistant HBV earlier.

 

Patients and Methods:

A total of 101 serum samples from 56 chronic HBV patients receiving Lamivudine and/or Adefovir were analyzed for the presence of antiviral-resistant mutations using both DRv2 and sequencing. The DRv2 involves reverse hybridization of PCR products onto strips coated with oligonucleotide probes. These probes can differentiate wild type vs. mutant sequences at codons 80, 173, 180 and 204 and at codons 181 and 236 of the HBV reverse transcriptase/polymerase, that are known to be associated with lamivudine and adefovir resistance, respectively. 

 

Results:

Complete concordance was observed for 573 (95%) of 606 analyzed codons (95% of samples analyzed for codon 80, 98% for codon 173, 87% for codon 180, 100% for codon 181, 90% for codon 204 and 97% for codon 236). Among the 33 discordant cases, DRv2 detected mutants while sequencing revealed wild type virus in 31 cases; sequencing of follow-up samples confirmed the presence of mutant sequences in all 27 cases with follow-up samples. In these 27 cases, DRv2 detected mutants earlier than sequencing by a mean of 6 months (range 2-11). In only 1 case sequencing detected mutant while DRv2 detected wild type virus; DRv2 of follow-up samples confirmed the presence of mutant sequence. Complete discordance was observed in only 1 case, where DRv2 showed wild type at position 80, whereas sequencing showed a STOP codon. The DRv2 assay is more rapid and amenable to high throughput compared to sequencing but it can only detect the presence of known drug-resistant mutations.

 

Conclusions:

Our study demonstrates that the INNO-LiPA HBV DRv2 can simultaneously detect the presence of Lamivudine and Adefovir-resistant mutations. The results of DRv2 show a high degree of concordance with sequencing and can detect mutants earlier, thus permitting prompt initiation of additional therapy.

 


Poster 970

Abstract ID: 64162

Category: J1O: Hepatitis B: Treatment

RECOGNITION OF MUTATED (G145R) HBsAg BY HEPEX-B (2 HUMAN MONOCLONAL ANTIBODIES) OFFERS A POTENTIAL TREATMENT FOR HBV IMMUNE ESCAPE PATIENTS.

R. Eren, XTL Biopharmaceuticals Ltd., Rehovot, Israel, D. Landstein, XTL Biopharmaceuticals Ltd., Rehovot, Israel, R. Kovjazin, XTL Biopharmaceuticals Ltd., Rehovot, Israel, O. Nussbaum, XTL Biopharmaceuticals Ltd., Rehovot, Israel, J. Ben- Porath, XTL Biopharmaceuticals Ltd., Rehovot, Israel, S. Shahar, XTL Biopharmaceuticals Ltd., Rehovot, Israel, T. Waisman, XTL Biopharmaceuticals Ltd., Rehovot, Israel, N. Haberman, XTL Biopharmaceuticals Ltd., Rehovot, Israel, D. Terkieltaub, XTL Biopharmaceuticals Ltd., Rehovot, Israel, S. Dagan, XTL Biopharmaceuticals Ltd., Rehovot, Israel

 

Introduction:

Long-term immunoprophylaxis with hepatitis B immunoglobulin (HBIG) is used for the prevention of recurrent hepatitis B virus (HBV) infection after liver transplantation. The most prevalent immune escape mutation from HBIG treatment is at position 145 (G145R) on the hepatitis B surface antigen (HBsAg). This mutation abolishes the neutralizing effect of HBIG, implying that neutralizing antibodies are directed to this region.

 

Purpose:

To test whether neutralizing monoclonal antibodies directed to different epitopes on HBsAg could be effective in decreasing selection pressure for the HBIGresistance mutation.

 

Methods:

HepeX-B consists of two human monoclonal antibodies (HumAbs): HBVAB17 that recognizes a conformational epitope and HBV-AB19 that recognizes a linear epitope on HBsAg. Epitope mapping was performed by reacting antibodies to overlapping 15-mer linear antigen-derived peptides. HBsAg harboring the G145R mutation was constructed and expressed in a baculovirus expression system.

 

Results:

The epitope to which HBV-AB19 binds was fully mapped. The core sequence of the epitope is CTKPTDGNC at positions 139-147. The cysteines surrounding this epitope are involved in binding, indicating that this is not a standard linear epitope. Changes at positions 141-145 resulted in a strong decrease of binding. The conformational epitope to which HBV-AB17 binds could not be mapped by this method. Western blot analysis showed that HBV-AB17 could recognize the wild type (wt) as well as the G145R mutated HBsAg whereas HBV-AB19 recognized only the wt antigen. Furthermore, FACS analysis of Hi-5 cells expressing the mutated HBsAg on their membrane demonstrated that only HBV-AB17 could bind to these cells. Cells expressing the wt HBsAg were recognized by both HumAbs. The neutralizing activity of HBV-AB17 and HBV-AB19 against virus harboring the G145R mutated HBsAg is being studied in a mouse model for HBV infection.

 

Conclusions:

The two HumAbs that comprise HepeX-B are directed against different epitopes on HBsAg. HBV-AB19 binds to a linear epitope that includes the sensitive site for the G145R immune escape mutation. Hence, HBV-AB19 did not recognize the G145R mutated HBsAg. The mutation at position 145 did not abolish the binding of HBV-AB17 to HBsAg. The binding to distinct epitopes on HBsAg and the fact that one of the HumAbs is not affected by the G145R mutation lower the probability of emergence of escape mutants due to HepeX-B therapy. Thus, HepeX-B could offer an alternative therapy to liver transplant patients who had escaped HBIG monotherapy.


Poster 971

Abstract ID: 65234

Category: J1O: Hepatitis B: Treatment

 

Impact of IFN on progression of liver disease in Hepatitis B “e” antigen (HBeAg) negative chronic hepatitis B (CHB) patients: a long term Italian multicenter A.I.S.F.study.

 

F. Oliveri, Uo Gastroenterologia e Epatologia, Az. Osp. Univ. Pisana, Pisa, Italy, M. Puoti, Clinica Malattie Infettive e Tropicali, AO Spedali Civili, Brescia, Italy, T. Santantonio, Clinica Malattie Infettive, Az. Osp. Policlinico Consorziale, Bari, Italy, P. Lampertico, Divisione di Epatologia, IRCCS Ospedale Maggiore Policlinico, Milano, Italy, G. Colloredo, Divisione Medicina, Policlinico San Pietro, Ponte San Pietro, Italy, G. Niro, Gastroenterologia Casa Sollievo della Sofferenza, San Giovanni Rotondo, Italy, G. Fattovich, Dipartimento di Gastroenterologia, Università di Verona, Verona, Italy, F. Morisco, Gastroenterologia, Dip. Scienza d. Alimenti, Univ. Napoli Federico II, Napoli, Italy, A. Marrone, Div. Medicina Interna ed Epatologia, Seconda Università di Napoli, Napoli, Italy, P. Costa, Divisione Malattie Infettive Ospedale "C. Poma", Mantova, Italy, M. Felder, Divisione di Gastroenterologia, Ospedale Centrale Bolzano, Bolzano, Italy, P. Cacciatore, Clinica Malattie Infettive, Università di Chieti, Chieti, Italy, A. Smedile, UO Gastroenterologia Ospedaliera, Osp S. Giovanni Battista "Molinette", Torino, Italy, M. R. Brunetto, Uo Gastroenterologia e Epatologia, Az Osp Univ. Pisana, Pisa, Italy

 

HBeAg negative CHB is a progressive disease with low spontaneous and drug induced resolution rates. We evaluated the Sustained Response (SR) rate to IFN and factors influencing both treatment and disease outcome in a large series of patients (pts).

 

We followed up prospectively 558 anti-HBe positive CHB pts (451 males and 107 females; mean age 41 y, range 12-66 y) from 13 Italian Centers. Patients were treated for at least 3 months (m) with alpha-IFN from 1985 to 2001: 437 (78%) pts experienced single IFN courses whereas 121 (22%) 2 or more courses. Mean follow up after end of treatment (EOT) 54 m (range 1-205 m). SR = normal ALT, HBV-DNA <10 pg/ml and IgM anti-HBc <0.2 IMx index for at least 12 m after EOT.

 

At baseline chronic hepatitis with cirrhosis was present in 150 (27%) pts, moderatesevere steatosis in 49 (9%). At first IFN treatment 319 (57,2%) pts achieved EOT response: 96 (17,2%) maintained response (SR), and 223 (40%) relapsed. The relapse occurred within 12 m after EOT in 183 (32,8%) pts, within 24 m in 24 (4,3%) pts, within 36 m in 10 (1,8%) pts and thereafter (till to 84 m) in the remaining 6. After relapse 14 (2,5%) pts developed persistently normal ALT and undetectable HBV-DNA (Persistent Remission after Relapse, PRR). The Long Term Response (LTR=SR+PRR) rate in naives (19,7%) was the same of that in retreated (19,8%) pts. At multivariate analysis (MVA) age younger than 40 y (p=0,011) and duration of treatment (p<0,000001) were independently associated with SR, that was observed in 6,8%, 15.9% and 30.7% of pts treated up to 6 m, 7-12 m and more than 1 y (mean 19 m) respectively. HBsAg clearance was observed in 47 (35%) of 134 LTR (anti-HBs seroconversion in 32). During follow up 43 (7,7%) pts developed End Stage Complications (ESC: jaundice, ascites, var.bleeding, encephalopathy) and/or Hepatocellular Carcinoma (HCC in 27, 4,8%). Four pts were transplanted because of HCC and 1 for terminal cirrhosis (t.c.); 14 (2,5%) died 15-100 m after EOT because of HCC (10), t.c.(3) and extraepatic tumor (1). Among 43 ESC pts 31 had baseline cirrhosis, only 1 was female. At MVA female sex, younger age, absent-mild steatosis, absence of cirrhosis and LTR to IFN were significantly and independently associated with a better outcome. In baseline cirrhotic pts, at MVA LTR was significantly associated with the absence of ESC except HCC.

 

Conclusions:

In HBeAg negative CHB standard IFN therapy warrants an overall LTR of 20%; a correct identification of SR requires a long term follow-up as relapse may occur late. LTR is associated with a better clinical outcome: in patients with cirrhosis IFN may avoid clinical decompensation, but not HCC.


Poster 973

Abstract ID: 66027

Category: J1O: Hepatitis B: Treatment

 

Unsuitability of Kaplan-Meier Estimates for determining long-term response rates in patients receiving treatment for chronic hepatitis B.

 

G. Cooksley, Royal Brisbane Hospital, Brisbane, Australia, G. K. Lau, Queen Mary

Hospital, Hong Kong, China, T. Piratvisuth, Songklanakarin Hospital, Songkla,

Thailand, M. Popescu, Roche, Basel, Switzerland, P. McCloud, Roche, Dee Why,

Australia

 

Background:

Kaplan-Meier (KM) estimates were originally developed to assess survival rates using death as the measured outcome. Recently, this statistical method has been used to assess the long-term efficacy of antiviral therapy for chronic hepatitis B (CHB). A basic assumption of the KM approach is that a patient who achieves an outcome (eg death) will always have that outcome whether they remain in the study or not. Since relapse occurs in CHB, we discuss the appropriateness of the KM approach in determining response to anti-HBV therapy.

 

Objective:

To compare HBeAg response rates obtained using the intent-to-treat (ITT) principle, which assumes that patients with missing data do not have a response, with rates obtained by KM estimation in patients enrolled in a large, randomized clinical trial.

 

Methods:

HBeAg-positive patients were treated for 48 weeks with 180 mg peginterferon alfa-2a (40KD) (PEGASYS®) once-weekly (qw) + oral placebo once-daily (qd), 180 mg peginterferon alfa-2a qw + 100 mg lamivudine qd, or 100 mg lamivudine qd. HBeAg seroconversion rates during treatment and up to 24 weeks post-treatment (week 72) were reanalyzed using the KM method.

 

Results:

HBeAg seroconversion rates at the end of treatment (week 48) in the ITT population were 27% [72/271] with peginterferon alfa-2a alone, 24% [64/271] with peginterferon alfa-2a + lamivudine and 20% [55/272] with lamivudine alone. Corresponding KM estimates of HBeAg seroconversion at week 48 (day 337) were 36% with peginterferon alfa-2a (+9% over ITT), 29% with combination therapy (+5% over ITT) and 24% with lamivudine (+4% over ITT). In the ITT population, HBeAg seroconversion rates 24 weeks post-treatment (week 72) were significantly higher with peginterferon alfa-2a alone (32% [87/271]; P<0.001) and peginterferon alfa-2a + lamivudine (27% [74/271]; P=0.023) than with lamivudine alone (19% [52/272]). KM estimates of HBeAg seroconversion rates at week 72 (day 504) were 43% with peginterferon alfa- 2a (+11% over ITT), 38% with the combination therapy (+11% over ITT) and 30% with lamivudine (+11% over ITT).

 

Conclusions:

Using data from this large, randomized study, estimated rates of HbeAg seroconversion generated using the KM method were substantially higher than the HBeAg seroconversion rates generated using an ITT analysis. These findings suggest that KM estimation is not an appropriate statistical method for measuring long-term efficacy of antiviral therapy in patients with CHB as it biases the data. Results generated using KM estimates should be interpreted with caution especially when the associated rates of relapse are not also analyzed.


Poster 974

Abstract ID: 66851

Category: J1O: Hepatitis B: Treatment

 

VIROLOGICAL BREAKTHROUGH IS NOT A DETRIMENTAL PHENOMENON IN PATIENTS WITH COMPENSATED CIRRHOSIS: RESULTS OF A LONG TERM LAMIVUDINE THERAPY

.

Y. Cakaloglu, Istanbul Medical Faculty Department of Gastroenterohepatology, Istanbul, NJ, Turkey, S. Kaymakoglu, Istanbul Medical Facutly Department of Gastroenterohepatology, Istanbul, NJ, Turkey, F. Akyüz, Istanbul Faculty of Medicine, Istanbul, NJ, Turkey, D. Ibrisim, Istanbul Medical Faculty Department of Gastroenterohepatology, Istanbul, NJ, Turkey, K. Demir, Istanbul Medical Faculty Department of Gastroenterohepatology, Istanbul, NJ, Turkey, D. Onel, Istanbul Medical Facutly Department of Microbiology, Istanbul, NJ, Turkey, E. Ahishali, Istanbul Medical  Facutly Department of Gastroenterohepatology, Istanbul, NJ, Turkey, B. Pinarbasi, Istanbul Medical faculty Department of Gastroenterohepatology, Istanbul, NJ, F. Besisik, Istanbul Medical Faculty Department of Gastroenterohepatology, Istanbul, NJ, Turkey, Z. Mungan, Istanbul Medical faculty Department of Gastroenterohepatology, Istanbul, NJ, Turkey, S. Badur, Istanbul Medical Faculty Department of Microbiology, Istanbul, NJ, Turkey, A. Okten, Istanbul Medical Faculty Department of Gastroenterohepatology, Istanbul, NJ, Turkey

 

Development of virological breakthrough during long-term lamivudine (LMV) therapy may result in fatal exacerbations in decompensated cirrhosis. The clinical results of virological breakthrough are still controversial in compensated or mildly decompensated cirrhosis.

 

Aim:

To evaluate the clinical significance of virologic breakthrough which develops during the long-term lamivudine therapy in patients with HBV related cirrhosis and the effects on clinical course and complications.

 

Material-Methods:

A hundred patients with HBV related cirrhosis (Child A 50, B 40, C

10) were enrolled and continuously treated with lamivudine 100 mg/day for 30±19 (12- 84) months. Baseline HBV DNA was positive (Hybridization, Digene) and ALT levels were high (>1.5XULN) in all patients. Normalization of ALT, negativity of HBV DNA (PCR<400-2000 copy/ml) and HBeAg seroconversion were accepted as on-therapy response. Reappearance of HBV DNA during the treatment was accepted as virologic breakthrough. Clinical course and complications were evaluated in patients with and without virologic breakthrough.

 

Results:

End of follow-up virologic and biochemical response rates were 56% and 58%, respectively (65.2% and 63.2% in HBeAg negative 70 patients and 32% and 46.4% in HBeAg positive 30 patients respectively- p<0.05). Virologic breakthrough developed in 25 patients (10 HBeAg positive, 15 HBeAg negative) in mean 33±15 months during LMV therapy. Of these 25 patients 11 (44%) had normal ALT. None of 14 patients with elevated ALT (2 had ALT level >10XULN) developed decompensation in liver disease. Although, CPT score did not significantly changed in total group, an important decrease was detected in patients without virological breakthrough. On the contrary virologic breakthrough was associated with a statistically significant increase in CPT scores (from 6.08±1.2 to 7.2±2.3, p<0.02). Also the frequency of complications of cirrhosis decreased with LMV treatment in comparison to pretreatment period. There was no significant difference in rate of complications between patients with and without virologic breakthrough. Thirteen patients (3 virologic breakthrough) developed hepatocellular carcinoma (HCC) in mean 29±19 months (Cumulative HCC incidence 5.6%). Seven patients (2 virologic breakthrough) died within 41±21 months (5 HCC, 1 AML-M4, 1 hepatic encephalopathy and spontanoeus bacterial peritonitis). Three patients underwent liver transplantation.

 

Conclusion:

Long-term LMV therapy results in significant improvement in laboratory and clinical parameters of cirrhosis. Virologic breakthrough is not associated with clinical deterioration in compensated and mildly decompensated cirrhotics.


Poster 976

Abstract ID: 67260

Category: J1O: Hepatitis B: Treatment

 

Factors associated with sustained virologic response 1 year after treatment with peginterferon alfa-2a (40KD) (PEGASYS®) monotherapy for HBeAg-negative chronic hepatitis B.

P. Marcellin, Hôpital Beaujon, Clichy, France, F. Bonino, IRCCS, Milan, Italy, G. K. Lau, Queen Mary Hospital, Hong Kong, China, P. Farci, Universita di Cagliari, Cagliari  Italy, C. Yurdaydin, University of Ankara, Ankara, Turkey, T. Piratvisuth, Songklanakarin Hospital, Songkla, Thailand, R. Jin, Beijing You An Hospital, Beijing, China, S. Gurel, University of Uludag, Bursa, Turkey, S. Hadziyannis, Henry Dunant Hospital, Athens, Greece, Z. Lu, Ruijin Hospital, Shanghai, China, M. Popescu, Roche, Basel, Switzerland

 

Background:

In patients with HBeAg-negative chronic hepatitis B (CHB), peginterferon alfa-2a (40KD) (PEGASYS®) alone or combined with lamivudine provides significantly higher response rates 24 weeks post-treatment compared with lamivudine alone. Multivariate analysis identified age, baseline ALT and baseline HBV DNA as significant predictors of virologic response (HBV DNA <20,000 copies/ml) to peginterferon alfa-2a when assessed 24 weeks post-treatment.

 

Objective:

To evaluate factors associated with sustained virologic response (defined as 48 weeks post-treatment response) to peginterferon alfa-2a monotherapy.

 

Methods:

HBeAg-negative patients (n=177) were treated for 48 weeks with 180 mg peginterferon alfa-2a (40KD) (PEGASYS®) once-weekly. HBV DNA was measured regularly during the 48 week post-treatment follow-up period (weeks 48, 52, 56, 60, 64, 72, 84 and 96). Analyses performed to identify baseline and on-treatment factors associated with post-treatment virologic response included the following variables: gender; race; age; bodyweight; baseline ALT; baseline HBV DNA; end of treatment HBV DNA; and HBV genotype.

 

Results:

Of 144 patients who had a virologic response (<20,000 copies/ml) to peginterferon alfa-2a monotherapy at the end of treatment, 89 had available data 48 weeks post-treatment. Of these patients, 49 (55%) sustained HBV DNA levels <100,000 cp/mL during the 48 week follow-up period [among these, 22 (25%) had HBV DNA levels between 20,000 and 100,000 copies/ml; and 27 (30%) had HBV DNA <20,000 cp/mL]. atients with sustained virologic response had higher mean ALT levels at baseline (94.7 IU/L) compared with patients with relapse (77.6 IU/L). Mean baseline HBV DNA levels were 7.18 and 6.99 log for sustained responders and relapsers, respectively. HBV DNA level at the end of treatment was not different in patients with sustained response and those with relapse (2.5 and 2.6 log, respectively). The rate of sustained virologic response according to HBV genotype was 60% [3/5], 43% [12/28], 64% [27/42] and 50% [5/10] for genotypes A, B, C and D, respectively (P=0.08 for comparison of genotype B vs C).

 

Conclusions:

In patients with HBeAg-negative CHB, a finite 48-week course of peginterferon alfa-2a was able to induce virologic response that was sustained 48 weeks post-treatment in more than half of the patients. There was no clear predictor of sustained response to peginterferon alfa-2a. However, there was a trend toward better response in patients with high ALT at baseline or who were infected with HBV genotype C. Baseline or end-of-treatment HBV DNA levels were not indicative of sustained virologic response.


Poster 977

Abstract ID: 61484

Category: J1O: Hepatitis B: Treatment

 

Efficacy of Tenofovir against HBV in HIV/HBV coinfected patients.

G. Klausen, Praxiszentrum Kaiserdamm, Berlin, Germany, A. Moll, Praxiszentrum Kaiserdamm, Berlin, Germany, J. Gölz, Praxiszentrum Kaiserdamm, Berlin, Germany, D. Schleehauf, Praxiszentrum Kaiserdamm, Berlin, D. Prziwara, Praxiszentrum Kaiserdamm, Berlin, S. Nzimegne-Gölz, Praxiszentrum Kaiserdamm, Berlin

 

Backround:

Tenofovir (TDF) is licensed for the treatment of HIV and is known to be active and potent as well against hepatitis B virus. Anyhow, there is still a lack of data about the efficacy of TDF against HBV in HIV/HBV coinfected patients. In our center we treated 37 HIV/HBV coinfected patients with TDF containing antiretroviral therapies since January 2002. 21 of those had a viral load of hepatitis B of more than 100000 IE/ml bevor the start of TDF. 10 of these patients had previously shown a clinically diagnosed resistance of HBV against Lamivudine (LAM).

 

Methods:

Retrospective analysis of the virological effectiveness of TDF against HBV in 21 HIV patients with highly replicative HBV coinfection, who were treated with a TDF containing antiretroviral therapy. Previous LAM resistance of HBV was defined as an increase of HBV of 2 log or more under treatment with LAM.

 

Results:

The 21 patients were all male and were in average 41 years old (range: 37-49). The average time on TDF was 27 month (range: 12-39) and the average viral load of HBV (VL) at baseline was 1.7x10log12 IE/ml (range: 2x10log5-1x10log13; N=21). At month 6 after start of treatment, the average VL was 743042 IE/ml(n=19), at month 12 it was 3769 IE/ml (n=17) and at month 24 the average VL was 230 IE/ml (n=15). The 10 patients with HBV resistance against LAM showed the following results: Baseline: VL 1,2x10log12 IE/ml (n=10), month 6: 33425 IE/ml (n=8), month 12: 7019 IE/ml (n=9), month 24: 386 IE/ml (n=8). In the reported patients we did not see any viral break through of HBV of more than 1 log under therapy with TDF so fare.

 

Conclusion:

According to our experience TDF is a long-term effective drug against HBV in HIV/HBV coinfeted patients with or without HBV resistance against LAM.


Poster 978

Abstract ID: 62282

Category: J1O: Hepatitis B: Treatment

Hepatitis B Virus Genotype B Is Associated With Better Response to Thymosin alfa-1 Therapy Than Genotype C.

 

R. Chien, Chang Gung Memorial Hospital and University, Keelung, Taiwan, Keelung,

Taiwan, Y. Liaw, Liver Research Unit, Chang Gung Memorial Hospital, Taipei, Taiwan

 

Background:

Hepatitis B virus (HBV) genotype has been reported to correlate with response to interferon (IFN) treatment in several studies. The relationship between HBV genotype and thymosin α 1 (T 1) treatment is unknown. We retrospectively examine HBV genotypes, precore and core promoter mutations in patients treated by T α 1 and analyze the correlation between complete response (CR; ALT normalization plus seroclearance of HBeAg and HBV-DNA by solution hybridization) of T α 1 therapy and HBV genotype.

 

Patients and Methods:

98 patients with clinicopathologically proven chronic hepatitis B were randomized allocated to three groups: 1) T6 group (n=32) received a 26-week course of T α 1 with a 1.6 mg subcutaneous injection two times a week; 2) T12 group (n=34) received the same regimen as T6 group, but T α 1 therapy extended for 52 weeks; 3) T0 group (n=32) served as a control and was followed up for 18 months without specific treatment. Retrospectively analyze the HBV genotype, precore and core promoter mutation using stored serum and correlated with CR.

 

Results:

There were 90 stored serum available for HBV genotyping from patients completing the treatment and follow-up study (29 in T6 group; 31 in T12 group and 30 in T0 group). Of them, 49 (54%) were genotype B and 41 (46%) were genotype C. Genotype C had a higher frequency of core promoter mutation. Stepwise logistic regression analysis showed that genotype (odds ratio [1] OR, 3.747; 95% confidence interval [1] CI,1.066- 13.170; P=0.039), precore mutation (OR, 6.285; 95% CI, 1.874-21.086; P=0.003) and T α -1 treatment (OR, 12.045; 95% CI, 2.220-65.354; P=0.004) as independent factors associated with CR. The CR of T α -1 was higher in patients with genotype B compared to patients with genotype C (52% of genotype B vs 24% of genotype C; P=0.036) and in patients with precore mutation (14/22 or 64% in responder vs 6/31 or 19% in nonresponder; P=0.002).

 

Conclusion:

Genotype, presence of precore mutation and T α-1 therapy were independent predictors to CR. Genotype B, compared to genotype C, is associated with a higher response rate to T α 1 therapy.


Poster 979

Abstract ID: 63873

Category: J1O: Hepatitis B: Treatment

 

Long-term experience of adefovir dipivoxil add-on therapy in chronic hepatitis B patients co-infected with HIV and lamivudine-resistant HBV : Absence of adefovir resistance mutation selection.

V. THIBAULT, Virology lab. CERVI - GH PITIE SALPETRIERE, PARIS, France, Y. BENHAMOU, GH PITIE SALPETRIERE, PARIS, France, M. VALANTIN, GH PITIE SALPETRIERE, PARIS, France, C. L. BROSGART, GILEAD, Foster city, CA, S. XIONG, GILEAD, Foster city, CA

 

Sequential monotherapy with lamivudine (LAM) and adefovir dipivoxyl (ADV) for chronic hepatitis B may quickly lead to the selection of resistant HBV. By contrast, a strategy based on add-on ADV therapy after LAM resistance emergence may be more effective.

 

We have analyzed the incidence of ADV resistance in HIV patients who have been treated for up to 5 years with a combination of ADV and LAM after development of LAM-resistance (LAM-R). In 2000, 35 HIV-patients were enrolled in an open label study of ADV add-on therapy after emergence of LAM-R HBV. In this follow-up study, all patients who presented residual HBV replication by PCR between year 4 and 5 of treatment were studied for the presence of HBV polymerase resistance mutations on serum samples. The HBV-RT was sequenced and compared to the sequences obtained before ADV introduction.

 

Among the 35 patients included in the initial cohort, 29 were still followed at year 4 and 14 had an undetectable viral load by PCR (<200 copies/mL). 15 had available samples with a viral load high enough to perform sequencing analysis. Median duration of add-on ADV of these 15 patients was 235 weeks and the median HBVDNA was 3.55 log10 cop./mL. 14/15 patients had constant fluctuating low viral loads between 2.3 (LLD) and 5 log10 cop./mL with no evidence of viral breakthrough. 10/15 patients had the same LAM-resistant pattern as their baseline sequence rtV173L/L180M/M204V (n=2); rtL180M/M204V (n=8) and no noticeable change was observed in their RT sequences along time. 2/10 of them had ADV replaced by tenofovir (TDF) when the background HIV treatment required adjustment. 4/15 patients lost their LAM-R mutations. For 2 of them failure of compliance was documented. For the 2 remaining, analysis of their HIV and HBV viral load kinetics clearly suggested lack of compliance. 1/15 patient had a viral breakthrough with a HBVDNA rise over 1 log10 on two consecutive samples. At that time, LAM-R mutations rtV173L/L180M/M204V were still present but no ADV mutation (rtA181V/T or rtN236T) or new conserved site mutations were identified; ADV was replaced by tenofovir and HBV DNA started to decline. Sequential monotherapy, after LAM–R breakthrough, could potentially increase the risk of subsequent HBV resistance selection. A strategy based on ADV add-on therapy seems very effective and is very unlikely to select for double ADV and LAM resistant variants, even in difficult to treat patients such those with HIV coinfection. The persistence of initially present LAM-R mutations, the low variability of the RT sequence and the absence of selection of further compensatory mutation indicate a strong pressure on a replication impaired virus.


Poster 981

Abstract ID: 64190

Category: J1O: Hepatitis B: Treatment

 

SEQUENTIAL ANTIVIRAL THERAPY LEADS TO THE EMERGENCE OF MULTIPLE DRUG RESISTANT HEPATITIS B VIRUS.

 

S. Villet, INSERM unit 271, Lyon, France, C. Pichoud, INSERM unit 271, Lyon, France, A. Ollivet, INSERM unit 271, Lyon, France, J. Villeneuve, Hôpital Saint-Luc , Division of Hepatology, Montreal, Canada, C. Trepo, INSERM unit 271, Lyon, France, F. Zoulim, INSERM unit 271, Lyon, France

 

We analysed the genotypic and phenotypic evolution of the viral quasi-species of 2 patients with chronic hepatitis B who failed antiviral therapy: one received successively lamivudine, add-on adefovir + lamivudine, followed by a lamivudine + adefovir + Hepatitis B immunoglobulins (HBIg) after liver transplantation, and the 2nd received IFN, lamivudine then entecavir. For genotypic analysis, a 1142 bp region of the polymerase gene encompassing the rt domain and overlapping the S gene was amplified by PCR for each sample, cloned and sequenced. For phenotypic analysis, an HBV replication-competent plasmid was constructed from HBV DNA isolated from patient serum (Durantel et al. Hepatology 2004). The rt gene from the selected clones previously sequenced was then subcloned in this vector. Transfection of these HBV mutants in hepatoma cell line led us to determine their replication efficiency and drug susceptibility.At baseline, all HBV genomes carried a wild-type (wt) rt gene but, for the first patient, 36 % harbored the P120S mutation within the S gene associated with vaccine escape. Following viral breakthrough to LAM monotherapy, a complex mixture of LAM-resistant HBV strains emerged, with the prevalence of the rtL180M+M204V mutant for the first patient and of the rtV173L+L180M+M204V mutant for the second. In vitro, all the tested mutants showed a 1000 fold resistance to LAM relative to wt HBV. Following the switch to ADV for the first patient, or to ETV for the second, the viral load dropped but rose again after 3 and 2 years of therapy respectively. During this rebound, we observed a complex mixture of LAM + ADV or ETV resistant strains. For the first patient, the selection of only one HBV mutant was finally observed. This mutant harbored the rtV173L+L180M+A181V+N236T mutations, a combination of resistance mutations to both drugs, and the sP120S mutation escaping to HBIg. Interestingly, the main LAM resistance mutation rtM204V disappeared from the viral population. The dominance of this complex rt mutant may be explained by our in vitro findings of a greater resistance to ADV+ LAM, and a level of replication equivalent to wt HBV. For the second patient, we observed a mixture of 3 mutants during rebound. All these mutants had the rtS202G mutation, not yet described, but also the rtL180M+M204V LAM resistance mutations despite the absence of LAM during ETV therapy. All these mutants were resistant to both LAM and ETV in vitro. Our results show the evolution of viral quasi-species towards the selection of mutants escaping to multiple selective pressures. This suggests that de novo combination therapy should be further evaluated to prevent drug resistance.


Poster 982

Abstract ID: 64393

Category: J1O: Hepatitis B: Treatment

 

HBV DNA levels at week 9 and number of HBV core gene mutations predict outcome of lamivudine/interferon á treatment in infancy-acquired chronic hepatitis B.

I. Kraslova-Carey, Institute of Liver Studies, London, United Kingdom (Great Britain), L. D'Antiga, Institute of Liver Studies, London, United Kingdom (Great Britain), M. Cavers, Institute of Liver Studies, London, United Kingdom (Great Britain), Y. Ma, Institute of Liver Studies, London, United Kingdom (Great Britain), S. Bansal, Institute of Liver Studies, London, United Kingdom (Great Britain), J. A. Byrne, Institute of Liver Studies, London, United Kingdom (Great Britain), G. Mieli-Vergani, Institute of Liver Studies, London, United Kingdom (Great Britain), D. Vergani, Institute of Liver Studies, London, United Kingdom (Great Britain)

 

Background:

During the immune tolerance phase that characterizes infancy-acquired chronic hepatitis B, hepatitis B virus (HBV) wild-type and variant strains coexist, although the wild-type dominates. During combination antiviral treatment most wild-type strains are eliminated and stable HBV core antigen variants become prevalent. The emergence of mutations within HBV core immunodominant epitopes may interfere with effective immune response and viral control.

 

Aim:

To evaluate the dynamic changes in serum HBV DNA levels during combined antiviral treatment in relation to the emergence of mutations within the HBV core gene and to study their contribution to treatment outcome.

 

Patients:

Twenty-three children (8 males, median age 10.2 years, range 2.9-16.8) with infancy-acquired hepatitis B (all HBeAg positive) were treated with lamivudine (3mg/kg/d) for 52 weeks and interferon α (5MU/m2 TIW) from week 9 for 44 weeks. According to treatment outcome, patients were divided into 5 responders (R) and 18 nonresponders (NR). Seven untreated HBeAg positive paediatric patients (3 males, median age 9.8 years, range 3.8-14.1) served as controls (C).

 

Methods:

HBV DNA levels were measured in serial serum samples by real-time TaqMan PCR before (treatment week 0– TW0), during (TW2, TW9, TW28, TW52) and after (follow-up week– FUW24) antiviral treatment. The number of HBV DNA copies per ng of genomic liver DNA was evaluated at baseline by the same method. Mutations within the HBV core gene were determined at baseline within the liver and in sequential serum samples by restriction fragment length polymorphism after nested PCR, and by direct sequencing at the same time-points as HBV DNA levels.

 

Results:

HBV DNA viral load in liver (log10 copies/ng of liver genomic DNA) and in serum (log10 copies/ml) is shown in the table (mean±SEM).

 

 

The decrease in the number of HBV DNA copies/ml in serum was significantly higher among responders (p=0.003) from TW9. The number of mutations within the HBV core gene was significantly higher in non-responders (4.38±0.6) than responders (1.25±0.5, p=0.023) at TW28. There was a strong correlation between serum HBV DNA levels and number of mutations in HBV core gene was found (p=0.002) among treated patients.

 

Conclusion:

A decrease of HBV DNA levels > 3.5 log10 copies/ml at TW9 predicts therapy outcome. The strong correlation between HBV DNA levels and the number of mutations within the HBV core gene suggests that variant strains contribute to virus control failure.


Poster 983

Abstract ID: 64955

Category: J1O: Hepatitis B: Treatment

 

FIVE YEARS OF SEQUENTIAL LAM TO LAM+ADV THERAPY SUPPRESSES HBV REPLICATION IN MOST HBEAg-NEGATIVE CIRRHOTICS, PREVENTING DECOMPENSATION BUT NOT HEPATOCELLULAR CARCINOMA.

 

P. Lampertico, IRCCS Maggiore Hospital, Milan, M. Vigano, IRCCS Maggiore Hospital, Milan, Italy, E. Manenti, IRCCS Maggiore Hospital, Milan, Italy, M. Iavarone, IRCCS Maggiore Hospital, Milan, Italy, G. Lunghi, IRCCS Maggiore Hospital, Milan, Italy, M. Colombo, IRCCS Maggiore Hospital, Milan, Italy