Sustained Viral Response Group Research Articles

Epigenetic Modulation in Viral Hepatitis C Infected Patients after Successful Therapy

Nowadays, hepatitis C-virus (HCV) represents a challenging liver condition for infected patients and health care systems worldwide. Thus, identifying new biomarkers for early diagnosis and therapy response prediction should be prioritized. Our study aims to identify new epigenetic markers in patients with HCV infection before and after successful direct-acting agents therapy (DAA) and correlate them with standard diagnostic tools. We analyzed blood samples from 12 healthy volunteers and 22 HCV infected patients, before and 3 months after DAA, by assessing: liver transaminases, alpha-fetoprotein, cryoglobulinemia, miR-7-1-3p, miR-21-3p, miR122-5p, miR-885-5p, miR-16-5p, and liver fibrosis (FibroScan®). The study groups were Ctrl (Control group), HCV (HCV infected patients before therapy), and SVR (sustained viral response group). miR-7-1-3p was up-regulated in SVR compared to HCV. No difference was observed between Ctrl and HCV. MiR-21-3p was up-regulated in SVR compared to Ctrl. MiR-122-5p was up-regulated in both, HCV and SVR groups, while miR-885-5p was up-regulated in HCV and down-regulated in SVR group. Moreover, miR-122-5p and miR-885-5p were correlated with liver cytolysis. Our results revealed an innovative panel of epigenetic biomarkers in the early stage of HCV infection and its variation after successful DAA therapy.

Possible Alterations in Appetite-related Molecules After the Elimination of Hepatitis C Virus.

Recent advances in antiviral treatment have achieved a sustained viral response (SVR) in over 95% of hepatitis C virus (HCV) infections. HCV elimination is suggested to improve several lifestyle-related factors; however, few studies have focused on dietary habit-/appetite-related factors. HCV-infected patients who received Daclatasvir/Asnaprevir (DCV/ASV) therapy were enrolled, and the changes in appetite-related molecules after antiviral therapy were assessed with a multiple cytokine-measuring system. Among 119 HCV-infected patients who received DCV/ASV treatment, 104 (87.3%) achieved an SVR. In the SVR group, DCV/ASV treatment improved several liver-related variables at 24 weeks after the completion of therapy. In patients with an SVR, the values of glucagon-like peptide 1 (GLP-1) and leptin were significantly increased at 24 weeks after completing direct-acting antiviral therapy. However, no significant change was observed in non-SVR patients, regardless of the receipt of direct-acting antiviral treatment. Gastrointestinal hormones related to the dietary habit and/or appetite may be influenced by HCV elimination.

Changes in serum levels of cytokeratin-18 fragments in patients with chronic hepatitis C under antiviral therapy

Background and Aims: Cytokeratin-18 is the known substrate for caspases, which are encountered during hepatic and pancreatic acinar apoptosis. Studies performed in recent years have indicated that the cleavage level of serum cytokeratin-18 (M30 antigen) is correlated with hepatic fibrosis and disease severity in both chronic hepatitis C and non-alcoholic steatohepatitis. It was shown that antiviral therapy in chronic viral hepatitis C patients significantly reduced hepatocellular apoptosis and cytokeratin-18 is accepted as a reliable marker of hepatocyte apoptosis. Our aim was to determine the correlation between the cytokeratin-18 level and treatment response in patients with chronic viral hepatitis C. Materials and Methods: Sixty patients with chronic viral hepatitis C were included in the study. A 48-week course of peginterferon- ribavirin therapy was given to appropriate patients. Hepatitis C virus RNA was measured at 0, 12, and 24 weeks at the end of therapy and 72 weeks. In addition, cytokeratin-18 levels were measured at 0, 24, and 72 weeks. Results: The mean age of 60 patients was 52±10.9 years. While 31 (51.6%) of patients were in the sustained viral response group, 29 (8.4%) of patients were in the non-sustained viral response group. It was determined that while the cytokeratin-18 level at week 0 in the sustained viral response group was 243±21, the cytokeratin-18 level at week 24 was 115±12 U/L and the difference between the level of cytokeratin-18 at weeks 0 and 24 were 127±209 U/L (p: .014). While the cytokeratin-18 level at week 0 in the non- sustained viral response group was 270±14; at week 24, the cytokeratin-18 level was 136±19 U/L and the difference between cytokeratin-18 levels at weeks 0 and 24 was 136±156 U/L (p > .5). At week 72, the cytokeratin-18 level in the sustained viral response group was 109±38 and the difference between weeks 0 and 72 was 134±215 (p < .002). Conclusion: In chronic viral hepatitis C patients, there was a correlation between sustained viral response and cytokeratin-18, which is a marker of apoptosis. During treatment, it was found that there was a relationship between sustained viral response and the decrease in cytokeratin-18 levels. Thisfinding indicates that cytokeratin-18 level monitoring may be used as a predictive marker of sustained viral response.

The correlation between interleukin-21 and the recurrence of chronic hepatitis B after entecavir discontinuation

Objective To study the correlation between interleukin (IL)-21 and the recurrence of chronic hepatitis B (CHB) in hepatitis B e antigen (HBeAg)-positive patients after discontinuation of entecavir (ETV). Methods A total of 112 HBeAg-positive CHB patients treated with ETV were enrolled. Serum levels of IL-21 at week 0, 12, 24 and 52 after ETV discontinuation were detected. The Cox regression model was used to analyze the correlation between IL-21 and the recurrence after ETV discontinuation. The receiver operating characteristic (ROC) curve was applied to determine the predictive value of IL-21 for CHB recurrence after ETV discontinuation. The expression differences of IL-21 were compared between sustained viral response (SVR) group and viral relapse (VR) group. The t test and Chi-square test were used for statistical analysis. Results The serum levels of IL-21 in SVR group at week 0, 12, 24 and 52 after ETV withdrawal were (60.0±10.8), (60.2±14.7), (60.6±19.5) and (61.2±15.4) ng/L, respectively, which were all significantly higher than those in VR group (47.5±10.7), (47.3±12.9), (46.9±12.2) and (46.4±12.8) ng/L, respectively (t=6.153, 4.926, 4.382 and 5.515, respectively, all P<0.01). The area under curve (AUC) was 0.811 (95%CI: 0.728~0.893, P<0.01) and the best cut-off value of serum IL-21 level was 49.8 ng/L. The recurrence rates of patients with serum IL-21 level ≥49.8 ng/L and <49.8 ng/L at time of ETV withdrawal was 25.4% (16/63) and 77.6% (38/49), respectively. The difference was statistically significant (χ2=30.027, P<0.01). The serum IL-21 level at the time of drug withdrawal (P=0.005), serum hepatitis B surface antigen (HBsAg) level at the time of HBeAg seroconversion (P=0.008) and age (P=0.016) were factors associated with CHB recurrence after ETV withdrawal by multivariate Cox model analysis. The serum levels of ALT, HBV DNA, HBeAg and HBsAg in SVR group were significantly lower than those of VR group at week 12, 24 and 52 after ETV withdrawal (t=-5.968, -7.691, -8.093; -3.047, -9.477, -28.900; -2.872, -10.424, -18.330; -4.633, -4.030 and -5.032, respectively; all P<0.01). Serum level of IL-21 was negatively correlated with HBsAg in SVR group after ETV withdrawal (r=-0.241, P<0.01), while positively correlated with HBsAg in VR group (r=0.286, P<0.01). Conclusions The serum IL-21 level at the time of drug withdrawal is associated with the recurrence after ETV discontinuation. IL-2l may play an important role as an immunomodulatory factor in maintaining virological responses in HBeAg-positive CHB patients after ETV withdrawal. Key words: Cytokines; Hepatitis B, chronic; Recurrence; Entecavir; Withdrawal

Serial changes in liver stiffness and controlled attenuation parameter following direct-acting antiviral therapy against hepatitis C virus genotype 1b.

Little information is available on the impact of direct-acting antiviral (DAA) therapy on changes in liver fibrosis and steatosis. Liver stiffness (LS) and controlled attenuation parameter (CAP) values were evaluated using transient elastography. The study subjects were 214 elderly patients infected with HCV genotype 1b who received 24-week daclatasvir and asunaprevir dual therapy. All patients of this retrospective study had no hepatocellular carcinoma before and during DAA therapy. LS and CAP were assessed before treatment (baseline), at end of treatment (EOT), and at 24, 48, 72 weeks (W) after EOT. The rate of sustained viral response (SVR) by daclatasvir and asunaprevir therapy was 91%. LS values for the entire group correlated with Fib-4 index at baseline (r = 0.565, P < 0.001). LS in both chronic hepatitis group (Fib-4 index <3.25) and cirrhosis group (Fib-4 index ≥3.25) decreased significantly at each time point compared with baseline (P < 0.001). Especially, a larger decrease in LS from baseline to EOT was seen in the cirrhosis group than chronic hepatitis group (P < 0.001). LS was also significantly lower in the SVR group at EOT, 24W, 48W, 72W compared with baseline (P < 0.001). Even in the non-SVR group, LS tended to be lower at EOT (P = 0.039), 24W (P = 0.009), 48W (P = 0.475), 72W (P = 0.033) compared with baseline. CAP increased significantly following the treatment from baseline to 48W post-EOT (P = 0.018). Our results showed significant improvement in LS in response to daclatasvir and asunaprevir dual therapy. In the other hand, there was a tendency that CAP increased from baseline.

Impact of interleukin 28B rs12979860 C/T polymorphism on severity of disease and response to treatment in hepatitis delta.

Pegylated-interferon alpha (Peg-IFN α) is the therapy most commonly used to treat chronic hepatitis delta virus (HDV) infection. In the present study, we planned to investigate effect of IL28B polymorphism on response to Peg-IFN α therapy and disease progression in patients with chronic HDV. A total of 47 patients who received Peg-IFNα therapy for at least one year were investigated. The patients were divided into three groups based on their response to treatment: sustained viral response (SVR) (32%), unresponsive (53%), and relapse (15%). The groups were compared in terms of age, gender, blood biochemistry (albumin, total bilirubin, lactic acid dehydrogenase, ALT, AST, ALP, GGT), complete blood count, HBeAg, HBsAg, HBV-DNA, HDV-RNA, IL28B genotypes (CC, CT, TT), and results of liver biopsy. Regarding the investigation of IL28B genotype, the prevalence of CC, CT, and TT showed no difference among the three groups. In the SVR group, the prevalence of CC was 53%, CT was 47%, but there was no patient with TT. In the unresponsive group, prevalence of CC was 52%, CT was 32%, and TT was 16%. In the relapse group, prevalence of CC was 43%, CT was 57%, but there was no patient with TT genotype. No significant difference was found among the groups with sustained response, no response, and relapse in terms of CC and CT polymorphisms (p>0.05). No relationship was found between IL28B rs12979860 polymorphism and response to treatment and disease severity in patients with chronic HDV infection.

Second hepatectomy for recurrent hepatocellular carcinoma achieved sustained virological response to interferon therapy for hepatitis C.

Interferon (IFN) improves the prognosis of HCV-related hepatocellular carcinoma (HCC) in patients. However, the effects of IFN therapy for second hepatectomy (Hx) for recurrent HCC have not been established. Subjects included 96 patients who underwent a second Hx for recurrence of HCV-related HCC. Forty-four patients received IFN therapy past or postoperatively of the first Hx. Twenty of those patients attained a sustained viral response (SVR). The other 24 were non-responders (NR) and 52 patients who had not received IFN therapy (non-IFN) were classified as the NR/non-IFN group. Overall survival (SVR group vs. NR/non-IFN group: 5-yr, 91.7 vs. 51.0%; p = 0.012) and disease-free survival (SVR group vs. NR/non-IFN group: 3-yr, 64.7 vs. 25.9%; p = 0.006) rates were significantly different in both groups. By multivariate analysis, NR/non-IFN therapy, was the independent risk factor for overall survival (p = 0.025) and disease-free survival (p = 0.006) after second Hx. SVR achieved past or postoperatively of the first Hx of HCV-related HCC significantly inhibits recurrence and consequently improves patient survival after second Hx for recurrent HCC. Patients with SVR to IFN therapy would be good candidates for second Hx for recurrent HCC.

Deep sequencing and phylogenetic analysis of variants resistant to interferon-based protease inhibitor therapy in chronic hepatitis induced by genotype 1b hepatitis C virus.

Because of recent advances in deep sequencing technology, detailed analysis of hepatitis C virus (HCV) quasispecies and their dynamic changes in response to direct antiviral agents (DAAs) became possible, although the role of quasispecies is not fully understood. In this study, to clarify the evolution of viral quasispecies and the origin of drug-resistant mutations induced by interferon (IFN)-based protease inhibitor therapy, the nonstructural-3 (NS3) region of genotype 1b HCV in 34 chronic hepatitis patients treated with telaprevir (TVR)/pegylated interferon (PEG-IFN)/ribavirin (RBV) was subjected to a deep sequencing study coupled with phylogenetic analysis. Twenty-six patients (76.5%) achieved a sustained viral response (SVR), while 8 patients did not (non-SVR; 23.5%). When the complexity of the quasispecies was expressed as the mutation frequency or Shannon entropy value, a significant decrease in the IFNL3 (rs8099917) TT group and a marginal decrease in the SVR group were found soon (12 h) after the introduction of treatment, whereas there was no decrease in the non-SVR group and no significant decrease in mutation frequency in the IFNL3 TG/GG group. In the analysis of viral quasispecies composition in non-SVR patients, major populations greatly changed, accompanied by the appearance of resistance, and the compositions were unlikely to return to the pretreatment composition even after the end of therapy. Clinically TVR-resistant variants were observed in 5 non-SVR patients (5/8, 62.5%), all of which were suspected to have acquired resistance by mutations through phylogenetic analysis. In conclusion, results of the study have important implications for treatment response and outcome in interferon-based protease inhibitor therapy. In the host, hepatitis C virus (HCV) consists of a variety of populations (quasispecies), and it is supposed that dynamic changes in quasispecies are closely related to pathogenesis, although this is poorly understood. In this study, recently developed deep sequencing technology was introduced, and changes in quasispecies associated with telaprevir (TVR)/pegylated interferon (PEG-IFN)/ribavirin (RBV) triple therapy and their clinical significance were investigated extensively by phylogenetic tree analysis. Through this study, the associations among treatment response, changes in viral quasispecies complexity in the early stage of treatment, changes in the quasispecies composition, and origin of TVR-resistant variant HCV were elucidated.

Genomic diversity of the genotype 1b hepatitis C virus open reading frame is correlated with outcomes of combined pegylated-interferon/ribavirin therapy in patients with chronic hepatitis C from Henan Province

To determine the role of hepatitis C virus (HCV) genotype 1 b genetic variation in the open reading frame for treatment outcomes of the pegylated-interferon/ribavirin (peg-IFN/RBV) combination therapy by examining patients from Henan Province with chronic hepatitis C (CHC). Thirty-seven treatment naive patients infected with HCV genotype 1b were included in the study. Prior to initiation of a 48-week course of peg-IFN/RBV therapy, peripheral blood was drawn for sequencing of the viral ORF 5'-half. Patients were assessed at the end of the 48 weeks of treatment and at a 6-month follow-up appointment. The patient data was stratified according to the status of sustained viral response (SVR) group and non-response (NR) and statistical analysis was performed to determine the correlation between detected genetic variations and treatment response status. Genetic variability in the ORF 5'-half was significantly higher among the individuals in the SVR group than among those of the NR group. Significant differences were found in the gene regions encoding p7, NS2 and NS3. For p7, the NR group had actual and expected frequencies of special mutation of 9.0 and 17.4 and total mutation of 77.0 and 68.6, while the SVR group had actual and expected frequencies of special mutation of 42.0 and 33.6 and total mutation of 124.0 and 132.4 (x2 =7.725, P =0.05). For NS2, the NR group had actual and expected frequencies of special mutation of 36.0 and 54.3 and total mutation of 270.0 and 251.7, while the SVR group had actual and expected frequencies of special mutation of 106.0 and 87.7 and total mutation of 388.0 and 406.3 (x2 = 12.16, P less than 0.01). For NS3, the NR group had actual and expected frequencies of special mutation of 49.0 and 53.4 and total mutation of 241.0 and 236.6, while the SVR group had actual and expected frequencies of special mutation of 81.0 and 76.6 and total mutation of 335.0 and 339.4 (x2 =6.745, P =0.043). The inter-patient genetic variations in the NS3 gene were concentrated in the protease domain. Furthermore, there was a strong correlation between HCV diversity in p7 and treatment outcome. The genetic data reported here provides strong support for the role of NS2, NS3 and p7 in antagonizing the peg-IFN/RBV response during the treatment of HCV infections. We conclude that higher inter-patient viral genetic diversity correlates with successful treatment and may modulate the efficacy of antiviral therapy in CHC patients of Henan.

Relationship between inosine triphosphate genotype and outcome of extended therapy in hepatitis C virus patients with a late viral response to pegylated‐interferon and ribavirin

It is not yet clear which factors are associated with the outcome of 72-week treatment with pegylated-interferon and ribavirin (RBV) in patients with chronic hepatitis C virus (HCV) infection. In 66 patients with HCV genotype 1 who had a late viral response (LVR) to 72-week treatment of pegylated-interferon and RBV, we examined the factors that determined the outcome, including single nucleotide polymorphisms of interleukin-28B and inosine triphosphatase (ITPA) genes. Thirty seven of 66 (56%) patients with LVR achieved a sustained viral response (SVR). The mean age of these 37 SVR patients was 55, compared with 61 in 29 relapsed patients (P = 0.009). Twenty six of 54 (48%) patients with the CC genotype and 11 of 12 (92%) with the CA/AA genotype of ITPA rs1127354 achieved SVR (P = 0.006). The SVR rates were 79%, 40%, 60%, and 33% in patients with undetectable HCV RNA on weeks 16, 20, 24, and 28 or later, respectively (P = 0.014). Finally, serum RBV concentration at week 44 of treatment was significantly higher in the SVR group (2651 ng/mL) than in the relapse group (1989 ng/mL, P = 0.002). In contrast, the rate of the interleukin-28B genotype was not different between the groups. Multiple regression analysis showed that age < 60 years, ITPA CA/AA genotype, and serum RBV concentration were significant independent predictive factors for SVR. Our findings elucidated the association of four factors, including ITPA genotype, with the outcome of 72-week treatment in LVR patients.

Impact of Interleukin-28B gene polymorphism [rs12979860] on Egyptian patients infected with hepatitis C virus genotype-4

Single nucleotide polymorphisms (SNPs) in the Interleukin (IL)-28B gene, namely rs12979860, could predict response to pegylated interferon-α-ribavirin (PR) therapy in hepatitis C virus genotype 1 (HCV-1)-infected patients. A similar role was investigated in a case-control study conducted on 93 Egyptian patients chronically infected with HCV-4 in comparison to 22 individuals with spontaneous HCV clearance and 70 healthy volunteers. The homozygous C allele genotype (CC) was associated with sustained viral response (SVR) to therapy compared with the homozygous T allele genotype (TT) and the heterozygous genotype (CT). In the SVR group, the response rate was statistically significantly higher in CC genotypes (58.6%) compared with CT/TT (20.3%). There was no correlation between SVR patients' genotypes and early response to therapy or HCV baseline viral load. Our findings describe how IL-28B SNP genotyping may guide appropriate selection of HCV-4-infected patients for PR therapy.

PMO-183 Role of IL28B polymorphism in prediction of response to therapy in patients with genotype 1 chronic hepatitis c infection

IntroductionPatients with chronic hepatitis C virus (HCV) infection have a variable response to antiviral therapy with pegylated interferon and ribavirin. Influences include age, gender, viral genotype, viral load, severity of...

Relation of pretreatment sequence diversity in NS5A region of HCV genotype 1 with immune response between pegylated-INF/ribavirin therapy outcomes

Hepatitis C virus (HCV) infection frequently persists despite substantial virus-specific immune responses and the combination of pegylated interferon (INF)-α and ribavirin therapy. Major histocompatibility complex class I restricted CD8(+) T cells are responsible for the control of viraemia in HCV infection, and several studies suggest protection against viral infection associated with specific HLAs. The reason for low rates of sustained viral response (SVR) in HCV patients remains unknown. Escape mutations in response to cytotoxic T lymphocyte are widely described; however, its influence in the treatment outcome is ill understood. Here, we investigate the differences in CD8 epitopes frequencies from the Los Alamos database between groups of patients that showed distinct response to pegylated α-INF with ribavirin therapy and test evidence of natural selection on the virus in those who failed treatment, using five maximum likelihood evolutionary models from PAML package. The group of sustained virological responders showed three epitopes with frequencies higher than Non-responders group, all had statistical support, and we observed evidence of selection pressure in the last group. No escape mutation was observed. Interestingly, the epitope VLSDFKTWL was 100% conserved in SVR group. These results suggest that the response to treatment can be explained by the increase in immune pressure, induced by interferon therapy, and the presence of those epitopes may represent an important factor in determining the outcome of therapy.

Hepatitis C Treatment Response in Patients with a Tobacco History: An Inner City Hospital Experience

Purpose: Tobacco smoking can acclerate liver fibrosis in those with viral hepatitis, alcohol abuse or obesity (1). Additionally, El-Zayadi et al. (2) showed reduced response to interferon monotherapy in smokers compared to non smokers. Pauly et al. (3) demonstrated reduced Sustained Viral Response (SVR) in smokers to combined interferon/ribavarin treatment in genotype 2/3 but not genotype 1 patients. Neither study commented on tobacco dose amounts or ex smoking history effect on SVR. We wished to ascertain if current or prior tobacco exposure impacted SVR rates in patients with genotype 1 or genotype 2/3 when treated with pegylated interferon/ribavarin. Methods: A Retrospective chart analysis of electronic records matching inclusion criteria was performed and patients divided into smokers, ex smokers, and non smokers at treatment time. Non SVR and SVR was evaluated in each group. Amount of tobacco smoked was measured in pack years (packs per day * years smoked). Results: Of 311 medical charts reviewed, 229 met the inclusion criteria. Pack year smoking amounts was documented in 158 charts. Figure 1 reveals genotype 2/3 had better treatment response than genotype 1. There was no real difference amongst groups when smoking history was taken into account. Amongst smokers the mean pack year smoking amount was 21 in the SVR group and 18.8 in the non SVR group (p=0.24).[254] Figure 1: SVR between G1 and G2/3 groups.Conclusion: This study suggests that current or prior tobacco smoking does not have a significant effect on SVR rates in hepatitis C patients treated with interferon/ribavarin. Additionally, response rates do not appear to depend on amount of cigarettes smoked. Larger studies need to be done to validate these findings. Smoking cessation should still be encouraged as part of health promotion efforts.Table 1: DemographicsTable 2: Odds ratio estimates for SVR vs. Non SVR rates for selected variables

PEG-IFNα/RBV Combination Therapy for Chronic Hepatitis C Patients Increases Serum Ferritin Level while It Improves Sustained Viral Response Rate

Objectives: We investigated the significance of serum ferritin levels in pegylated interferon (PEG-IFN) and ribavirin (RBV) combination therapy for chronic hepatitis C (CHC) and examined its correlation with serum alanine aminotransferase (ALT) levels during therapy and response to the therapy. Methods: A total of 175 patients with CHC received the combination therapy. Correlations between serum ferritin levels and serum ALT levels at 12 and 24 weeks of therapy were examined. Differences in serum ferritin levels during therapy between patients with sustained viral response (SVR) and non-SVR were also examined. Results: Only 24 (13.7%) and 20 (11.4%) patients showed elevated serum ALT levels (≧70 IU/l) at 12 and 24 weeks of therapy, respectively. There was no correlation between serum ferritin levels and ALT levels. Ninety-five (54.3%) of 175 patients achieved SVR. Serum ferritin levels increased dramatically in both SVR and non-SVR groups after starting the therapy and were significantly higher in the SVR group throughout the therapy. Conclusions: Serum ferritin level increases during PEG-IFN and RBV combination therapy; however, it did not correlate with either serum ALT level or the total dose of RBV. Higher serum ferritin levels during combination therapy appear to be associated with favorable therapeutic response.

Predictors of alpha‐fetoprotein elevation in patients with chronic hepatitis C, but not hepatocellular carcinoma, and its normalization after pegylated interferon alfa 2a–ribavirin combination therapy

Elevated serum alpha-fetoprotein (AFP) levels are noted in patients with chronic hepatitis C (CHC) without hepatocellular carcinoma (HCC). The change in AFP levels after treatment with pegylated interferon and ribavirin (Peg-IFN/RBV) combination therapy is still unknown. The aim of this study was to investigate the predictors of elevated serum AFP in patients with CHC, and its change after Peg-IFN/RBV therapy. A total of 123 patients, intended to receive pegylated interferon alfa-2a plus ribavirin therapy, were enrolled. Eighty-three patients had complete treatment and received follow up for and additional 24 weeks. The factors that may affect the elevation of pretreatment AFP and the normalization of post-treatment AFP were determined. The mean AFP level was 18.5 +/- 63.0 ng/mL (range, 1.3-676.0 ng/mL); 41 (33.3%) of the 123 patients had elevated serum AFP (more than 10 ng/mL) at baseline. A multivariate logistic regression analysis disclosed that older age (odds ratio [OR], 1.093; 95% confidence interval [CI], 1.015-1.177; P = 0.018), more advanced METAVIR fibrosis stage (OR, 5.237; 95% CI, 1.244-22.037; P = 0.024), a higher aspartate aminotransferase (AST) level (IU/L) (OR, 1.020; 95% CI, 1.008-1.033; P = 0.001), and lower platelet count (x10(9)/L, OR, 0.985; 95% CI, 0.968-0.994; P = 0.003) were independent determinants of pretreatment AFP elevation. After treatment, 72 of 83 (86.7%) cases were found to have normal post-treatment AFP levels (<10 ng/mL) at the end of follow up (EOF). Post-treatment negativity of the chronic hepatitis C virus (HCV)-RNA (OR, 10.014; 95% CI, 1.000-100.329; P = 0.050) and the post-treatment platelet count (x10(9)/L) (OR, 1.025; 95% CI, 1.001-1.050; P = 0.040) were associated with normal AFP at EOF. AFP progressively decreased with significant differences starting from the 12th week after treatment to the end of treatment, and was lowest at the EOF date for the sustained viral response (SVR) group. On the contrary, the non-SVR group did not have an AFP change during and after treatment. Older age, low platelet count, higher AST levels, and advanced fibrosis predisposed chronic hepatitis C patients without HCC to have elevated serum AFP levels. After Peg-IFN/RBV combination therapy, a higher platelet count and HCV viral eradication were determinants of normal AFP at EOF. Serial AFP levels decreased after treatment, presenting in a time-dependent manner, specifically for the SVR group.

Mutations in the NS5B region of the hepatitis C virus genome correlate with clinical outcomes of interferon‐alpha plus ribavirin combination therapy

Combination treatments of interferon-alpha (IFN) and ribavirin (RBV) are more effective than those of IFN alone in hepatitis C virus (HCV) infection. However, mechanisms of the action of the combination regimen are not well understood. To elucidate the viral genetic basis of IFN plus RBV combination therapy, genetic variabilities of HCV-1b were analyzed. We performed pair-wise comparisons of full-length HCV genomic sequences in three patients' sera before and after initiation of IFN plus RBV treatment. Subsequently, we analyzed amino acid sequences of the NS5B region, which codes for the viral RNA-dependent RNA polymerase, and compared these with the outcomes of the therapy in 81 patients. Analysis of the entire HCV sequence in patients who received IFN plus RBV therapy did not show consistent amino acid changes between before and after the initiation of the therapy. NS5B sequence analyses revealed that mutations at positions 300-358 of NS5B, including polymerase motif B to E, occurred more frequently in a group of patients exhibiting a sustained viral response (SVR) or an end-of-treatment response (ETR) compared with a group of patients exhibiting a non-response (NR). Closer examination revealed that mutations at aa 309, 333, 338 and 355 of NS5B occurred significantly more frequently in the SVR plus ETR group than in the NR group (P = 0.0004). Multivariate analysis showed that the number of mutations at these four sites was an independent predictor of SVR plus ETR versus NR. Particular amino acid changes in the NS5B region of HCV may correlate with outcomes of IFN plus RBV combination therapy.