Session Title: Parallel Session 30: Understanding and Enhancing HCV Treatment II

 

205. 5-Year Follow-Up of Clinical and Histological Changes in Patients With HCV Infection and Initially Mild Liver Disease.

M. Simonova; K. Katzarov; D. Takov; E. Nakov; N. Tomov; D. Z. Alexieva; G. N. Vasilev; E. Belokonski; N. Vladov

 

Introduction:

Many patients with HCV infection initially present with normal ALT and F0 or F1 on liver biopsy. Whether this is an early transitional stage in the course of progressive disease or a separate and stable condition is currently debated. This study aimed to assess clinical and histological changes during 5 years of follow-up in patients with HCV infection, initially mild liver disease, and normal ALT and F0/F1 on biopsy.

 

Patients and Methods:

From Jan 1999 until Jun 2001, 280 patients (aged 18-65 years) with HCV infection (HCV RNA-positive), normal ALT for 3 months, mild chronic hepatitis F0/F1 (Metavir), and no co-infections with HBV/HIV were selected for the study. Patients were separated into 3 groups according to age: Group 1, 18 to 33 years old (n=160); Group 2, 34 to 49 years old (n=105); Group 3, 50 to 65 years old (n=15). 112 of the patients in group 1 and 2 were addicted patients (80 on methadone maintenance therapy; 32 with active intravenous drug use). ALT was measured every 4 months and, if elevated, every month. Quantitative HCV PCR was conducted every year. A second liver biopsy was performed after 5 years. Antiviral therapy was considered if ALT remained elevated for more than 6 months.

 

Results:

196 patients completed the 5-year follow-up period. Most patients lost to follow-up were addicted patients. In Group 1 (n=106 finished study): ALT elevation was observed in 70/106 patients (66%); 31 patients started antiviral therapy; second liver biopsy showed fibrosis progression in 51/106 patients (48%) – F0 to F1 in 22/51, F1 to F2 in 28/51, and F1 to F3 in 1/51. In Group 2 (n=75 finished study): ALT elevation was observed in 37/75 patients (50%); 16 patients started antiviral therapy; fibrosis progression was found in 35/75 patients (46%) – F0 to F1 in 10/35, F1 to F2 in 20/35, and F1 to F3 in 5/35. In Group 3 (n=15 finished study); ALT elevation was observed 3/15 patients (20%); no antiviral therapy was required; fibrosis progression was found in 4/15 patients (26%) – F0 to F1 in 3/15, F1 to F2 in 1/15. ALT elevations and fibrosis progression were observed more frequently in addicted patients than in non-addicted patients.

 

Conclusion:

Mild chronic hepatitis C was rarely found in patients aged 50 to 65 and was associated with stable course of disease. Younger patients more frequently displayed signs associated with progressive disease (clinical and histological), especially in addicted patients, thereby suggesting the need for frequent ALT monitoring and antiviral treatment.

 


206. Identification of novel CTL epitopes in hepatitis C virus by genome-wide computational scanning and a rational design of peptide vaccine.

T. Mashiba; K. Udaka; Y. Hiasa; Y. Hirachi; Y. Satta; S. Kataoka; M. Kohara; M. Onji

 

Background/Aims:

Hepatitis C virus (HCV) remains a serious threat due to its persistence. Moreover, therapy of chronic HCV infection is difficult, especially that of genotype 1b. Targeted induction of cytotoxic T lymphocytes (CTLs) against infected cells represents a promising strategy for viral containment. However, finding HLA-binding peptides and screening these peptides for T-cell responses is a laborious, expensive and time-consuming process. Moreover, the emergence of epitope variants that not only escape antigen presentation, but also persist as a quasi-species is imminent. To overcome these problems, we have developed a query learning algorithm based on hidden Markov models (HMMs), a computational method to analyze the specificity of MHC class I-binding peptides by exploiting a data mining technique. Using this strategy, attempts were made to identify several HCV epitopes that can induce CD8+ T-cell responses effectively.

 

Methods:

The entire genome of HCV genotype 1b was examined for HLA-A*2402-binding peptides, the most frequent allele among Asians and with a frequency of around 10% in Western countries. High-scoring peptides were synthesized and subjected to HLA-binding assays. Peripheral blood mononuclear cells (PBMCs) obtained from patients infected with HCV genotype 1b and healthy individuals were cultured in the presence of peptides, and tested for cytolysis by 51Cr release assay. Peptide-specific cell lines were further examined for recognition of a HepG2 cell line stably expressing HCV genotype 1b full-genome (tamoxifen-treated RzM6).

 

Results:

We identified 32 high-binder peptides (log Ka>5.5) from an HCV genome and its variants using HMM methods. Peptide-specific CTLs could be induced from patients and healthy individuals for 12 of the 15 peptides tested. Peptide recognition was restricted by HLA-A24 molecule. Tamoxifen-treated RzM6 cells were lysed by most peptide-specific cell lines established, indicating the recognition of naturally processed peptides on the cell surface. Once stimulated optimally, these peptide-specific cell lines could tolerate most of the single amino acid substitutions present in variant viruses. Finally, HLA-A*2402-binding epitope peptides were found to have been evolutionarily conserved among Japanese subjects, for whom the 60% bearing this allele indicate the feasibility of peptide-based immunotherapy.

 

Conclusions:

An efficient method to identify CTL epitope peptides was introduced to design a peptide-based vaccine for HCV genotype 1b. Improved ease of identifying multiple epitopes facilitates the design of peptide-based immunotherapies that are robust against highly variable targets like HCV genotype 1b.

 


207. Influence of the IL-6 gene polymorphism on response to HCV therapy in HCV- and HCV/HIV-infected patients.

J. Nattermann; M. Vogel; R. Bruno; G. Ahlenstiel; A. Baumgarten; G. Klausen; M. Schulz; A. Iwan; H. Nischalke; T. Sauerbruch; J. Rockstroh; U. Spengler; a. for the Kompetenznetz HIV/AIDS

 

Background/Aims:

Pegylated interferon/ribavirin combination treatment is the current therapeutic standard for hepatitis C in HIV infection. However, individual treatment outcome is varied with prognostic factors poorly understood in this setting. IL6 may affect interferon antiviral efficacy via shared induction of STAT (activation of signal transducers and activators of transcription) proteins and shows a functionally relevant single nucleotide polymorphism in its gene. Here, we studied whether the IL6- poly-morphism affects HCV antiviral therapy in HIV/HCV co-infection.

 

Methods:

Using the cytokine genotyping tray (One Lambda, CA, USA) we determined IL6 genotypes in 115 HCV/HIV co-infected patients comprising 36 and 79 patients with acute and chronic HCV infection, respectively. In addition, 75 patients with chronic HCV monoinfection were studied. Patients were stratified according to IL-6 genotypes as high (HP; IL6 +174G/G, and +174G/C) and low (LP; IL6 +174C/C) IL-6 producers. End of treatment (ETRs) and sustained virological responses (SVRs) to standard treatment were compared in the resulting subgroups. Furthermore, induction of total and phosphorylated STAT3 expression by interferon was determined in transiently HCV core-transfected HUH7 cells which were cultured with varied concentrations of IL6.

 

Results:  Overall, ETR and SVR were achieved in 85/115 (74%) and 56/106 (53%) of HCV/HIV co-infected patients, respectively. IL6 HP had significantly better ETR (79% vs. 50%; p=0.007) rates and SVR rates (58% vs. 32%; p=0.027) than IL6 LP. This effect was also consistently seen when patients with acute and chronic hepatitis C were analyzed separately. Multivariate analysis confirmed the IL6 HP state as an independent positive predictor for ETR (odds ratio: 4.6; p=0.003) and SVR (OR: 3.1; p=0.035). Likewise, HCV monoinfected IL-6 HP had a significantly better SVR than patients carrying the LP genotype (54% vs 27%, p=0.02). In vitro, addition of IL-6 to HCV core transfected HUH7 cells resulted in a dose-dependent activation of STAT3.

 

Conclusions:

Taken together, our data suggest that the IL6 gene polymorphism affects treatment responses in HIV/HCV co-infection, probably via improved STAT signaling in the IL6 HP subgroup of patients.

 


208. A randomized, controlled, open-label study of peginterferon alfa-2a (40KD) (PEGASYS®) plus ribavirin (COPEGUS®) for 16 vs. 24 weeks in patients with genotype 2 hepatitis C infection.

M. Yu; C. Dai; J. Huang; N. Hou; L. Lee; M. Hsieh; C. Chiu; Z. Lin; S. Chen; M. Hsieh; L. Wang; W. Chang; W. Chuang View Pres.

Background:

The recommended treatment for patients with HCV genotype 2 infection is a pegylated interferon plus ribavirin for 24 weeks. However, SVR rates of >80% with peginterferon alfa-2a (40KD) plus ribavirin in genotype 2/3 patients have prompted interest in whether even shorter treatment duration can yield similarly high SVR rates. Therefore, we determined whether the efficacy of 16 weeks’ therapy was comparable to the standard 24-week treatment course in HCV genotype 2 patients.

 

Methods:

In a controlled, multicenter, open-label study in Taiwan, 150 treatment-naďve patients with HCV genotype 2 infection were randomized (1:2) to 16 weeks (n=50) or 24 weeks (n=100) peginterferon alfa-2a (40KD) 180 μg/week plus ribavirin 1000/1200 mg/day, with a follow-up period of 24 weeks. The primary endpoint was sustained virological response (SVR; undetectable HCV RNA [<50 IU/mL] after 24 weeks’ untreated follow-up). SVR was also determined in patients with and without a rapid virological response (RVR; undetectable HCV RNA after 4 weeks).

 

Results:

Baseline characteristics were similar in the two groups (Table). Overall, an SVR was achieved in 94% and 95% of patients receiving 16 and 24 weeks’ treatment (Table). Patients with an RVR had a significantly higher SVR rate than patients without an RVR in both the 16- and 24-week treatment arms. For patients without RVR, the mean dose of ribavirin from week 5 to 16 of treatment was significantly lower in non-responders than in responders (11.3 ± 2.5 vs. 16.1 ± 0.8 mg/Kg/day; p=0.034 [Mann-Whitney test]). Multivariate logistic regression analysis in all patients showed that an RVR and age were independently associated with an SVR. Both treatment arms were equally well tolerated. The incidence of alopecia was significantly higher in the 24-week group (49%) than in the 16-week group (20%, p=0.001).

 

Conclusion:

In this study, high SVR rates (>94%) were seen with both 16 and 24 weeks’ of peginterferon alfa-2a (40KD) plus ribavirin 1000/1200 mg/day in genotype 2 patients from Taiwan. This probably reflects the absence of negative prognostic factors in this patient population and the rapid rate with which patients responded (>86% RVR rate in both arms). We were unable to detect a difference in SVR rates with 16 and 24 weeks’ treatment among the 150 patients included in our trial.

 

Baseline characteristics

16 wks (n=50)

24 wks (n=100)

Male, n (%)
Age, yrs
BMI, kg/m2
HCV RNA, log IU/mL
Advanced hepatic fibrosis, F3/4, n (%)

32 (64)
50.8
25.1
4.98
11 (22)

58 (58)
49.9
24.8
4.88
20 (20)

Efficacy, n (%; 95% CI)

RVR

43 (86; 76–96)

87 (87; 80–94)

SVR - overall
SVR - Pts with an RVR, n (%)
SVR - Pts without an RVR, n (%)

47 (94; 87–100)
43/43 (100)a
4/7 (57)

95 (95; 91–99)
85/87 (98))b
10/13 (77)

 

ap=0.015, bp=0.002 vs. pts without an RVR

 


209. Improving the clinical relevance of pre-treatment viral load as a predictor of sustained virological response (SVR) in patients infected with hepatitis C genotype 1 treated with peginterferon alfa-2a (40KD) (PEGASYS®) plus ribavirin (COPEGUS®). 

S. Zeuzem; M. W. Fried; K. Reddy; P. Marcellin; M. Diago; A. Craxi; P. Pockros; M. Rizzetto; D. Berstein; M. L. Shiffman; A. Lin; S. J. Hadziyannis

 

Background:

Baseline HCVRNA is an important predictor of SVR in CHC genotype 1 (G1) pts and may be a useful tool to select an appropriate treatment strategy, particularly in pts with high viral load (VL). In the past, pts have been classified as “high” or “low” VL using a cut-off of 2x106 copies/mL. This value was based on data generated using conventional interferon-based regimens or PEG-IFN monotherapy. Furthermore, when HCVRNA assays were standardized, 2x106copies/mL translated (depending on assay) to either 600 or 800x103 IU/mL. Here, an exploratory analysis was performed to identify a baseline HCVRNA cut-off that most effectively differentiates between high and low probability of SVR in pts receiving current standard-of-care.

 

Methods:

Naive, HCV G1 pts(n=568) receiving PEG-IFNα-2a (180µg/week) plus RBV (1000/1200mg/day) in 2 phase III trials (NEJM,2002;347:975; Ann Int Med,2004;140:346). The likelihood of achieving an SVR (undetectable HCVRNA [<50IU/mL] at 72wks), was estimated as a function of baseline VL strata using a generalized additive multiple logistic regression model taking into account discrete (gender, race, cirrhosis status) and continuous variables (age, weight, baseline VL and pretreatment ALT quotient). Also, the receiver operating characteristic curve (ROC) was plotted.

 

Results (Table):

In the logistic regression model, which included all predictors listed above, HCVRNA was a strong independent predictor of SVR (p<0.001). However, this effect was found to be non-linear, indicating that above a cut-off of 5.6 log10 IU/ml the contribution of VL in predicting SVR is minimal. Based on this model and the ROC analyses, the baseline level that most effectively differentiated between a high and low probability of SVR was 5.6 log10 (~400x103) IU/mL. Using this cut-off, the SVR rate in pts with VL ≤400x103 IU/mL was 70%, compared with 43% in pts with baseline VL >400x103 IU/mL (Table). SVR rates in pts with high VL were similar across all three cut-off definitions, reflecting that pts in the VL category 400–800x103 IU/mL had SVR rates similar (43%) to those in pts with VL >800x103 IU/mL.

 

Conclusions:

This analysis shows that a baseline HCV RNA level of approximately 400x103 IU/mL is optimal for use as a cut-off point to best discriminate between low and high VL, based on the probability to achieve an SVR in G1 pts when treated with PEG-IFNα-2a + RBV for 48 wks. Use of this cut-off point will allow treatment optimization in G1 pts.

 

SVR in pts with HVL and LVL using different cut-offs

 

 

≤400 vs >400 x 103 IU/mL

≤600 vs >600 x 103 IU/mL

≤800 vs >800 x 103 IU/mL

n/N

90/129

189/439

112/177

167/391

156/206

156/362

SVR (%)

70

43

63

43

60

43

 


210. Platelet serotonine levels as predictor for the need of antidepressant treatment before and during chronic hepatitis C and interferon-alfa treatment. 

M. Schaefer; A. Hinzpeter; G. Janssen; M. Pich; M. Schwaiger; R. Sarkar; A. Friebe; R. Uebelhack; A. Heinz; T. Berg; L. Franke

Aims:

Severe psychiatric side effects such as depression may occur during treatment of chronic hepatitis C with interferon-alfa (IFN-alfa). Antidepressants have been shown to be effective for the acute treatment or for the prevention of IFN-alfa associated depressive symptoms. However, not all patients will develop depressive symptoms. Thus biological markers are needed that predict the need of antidepressant treatment before or during antiviral therapy of chronic hepatitis C. We therefore measured platelet serotonine (5-HT) before and during treatment of chronically HCV-infected patients with pegylated interferon-alfa and ribavirin.

 

Method:

5-HT platelet content was measured in 87 HCV-infected patients before and during treatment with pegIFN-a and ribavirin. Patients were divided in four groups:

1.     patients without depression before and during antiviral treatment

2.     patients with depression before treatment who got an antidepressant (AD) pre-treatment,

3.     patients who received AD after 4 weeks and

4.     patients who received AD after 12 weeks of antiviral therapy.

 

Findings:

Platelet serotonine concentrations decreased significantly during 6 months of IFN-alfa treatment in all groups. Patients without depression and AD treatment (group 1) had significant higher serotonine concentrations at all points of measurement. Patients who were depressed at baseline (group 2) had significant lower 5-HT concentrations compared to controls (p=0.024). In contrast 5-HT platelet concentrations in patients who developed depression after 4 or 12 weeks (group 3 and 4) did not differ significantly from controls at baseline. However, group 3 an 4 showed significant lower 5-HT concentrations if compared to controls after 4 weeks (p=0.02 and p=0.001 respectively) and after 3 months (p=0.011 and p=0.002 respectively). After 6 months only group 4 who started with AD after 3 months of treatment had significant lower 5-HT concentrations if compared to controls (p=0.013).

 

Conclusions:

Platelet serotonine concentration is significantly associated with depression during chronic hepatitis C infection and/or antiviral treatment. Monitoring platelet 5-HT might help to estimate the need and the best time point of starting antidepressant treatment in patients with CHC and antiviral therapy with peg IFN-alfa and ribavirin.