- Email: [email protected]
S1933 The Difference of Hepatitis C Viral (HCV) Internal Ribosomal Entry Site (Ires) and Picornaviral Ires-Mediated Translation Inhibited By Oligonucleotides
mediated by the activation of p38 MAPK pathway. In contrast, when combined with IFNa, nicotine disturbed the antiviral effects of IFN on HCV replication, involving PI-3K/Akt signaling pathway. In conclusion, treatment with nicotine possesses the unique effects on innate cellular defense pathways through the activation of cellular MAPK.
HBeAg. HBV-DNA was positive in all samples. Detection of a 540bp PCR product indicated positive sample for HBV DNA. RFLP analysis and SSP results showed that all patients had genotype D and all isolates that were sequenced were classified as genotype D. So, results were consistent to sequencing. The phylogenetic tree was constructed using phylips software and its result showed that the isolates are derived from the same evolutionary tree. Conclusion: Genotype D is the most prevalent HBV genotype in north-east Iranian patents. PCR-RFLP can use to detect of HBV genotyping in clinical practice.
S1933 The Difference of Hepatitis C Viral (HCV) Internal Ribosomal Entry Site (Ires) and Picornaviral Ires-Mediated Translation Inhibited By Oligonucleotides Xiaoyu Li, Steffen Mueller, Eckard Wimmer
S1936
Background and Aims: This phase 2b, active-controlled study evaluated the efficacy, safety, and HRQOL of albinterferon alfa-2b (alb-IFN), a novel recombinant protein comprising IFN alfa-2b genetically fused to human albumin, in combination with oral ribavirin in IFN treatment-naïve patients with genotype 1, chronic hepatitis C (CHC). HRQOL was assessed by changes in SF-36 v2 and missed workdays due to CHC in patients treated with albIFN vs peginterferon alfa-2a (PEG-IFN). Methods: 458 patients were randomized to 4 subcutaneous treatment groups: PEG-IFN 180 mcg qwk, and alb-IFN 900 and 1200 mcg q2wk, and 1200 mcg q4wk. Primary efficacy endpoint: sustained virologic response (SVR). SF-36 was evaluated at baseline, wk 4, 12, 24, and 48 on treatment, and wk 4, 12, and 24 post-treatment. Patient disability was evaluated by number of missed workdays or days with impaired activity. Missing data were handled using the last observation carried forward. Results: By intention-to-treat analysis, SVR rates were 58.5% for alb-IFN 900 mcg, 55.5% and 50.9% for 1200 mcg q2wk and q4wk, respectively, and 57.9% for PEG-IFN (P = not significant). Overall, alb-IFN had less impact on the physical and mental domains of SF 36 during treatment vs PEG-IFN. Reductions in SF-36 scores occurred by wk 4-12, consistent with the occurrence of most treatment-associated adverse events. At wk 12, there was significantly less worsening in SF-36 physical (P = .04) and mental (P = .02) component scores, as well as in role-physical (P = .02), bodily pain (P = .003), vitality (P = .02), socialfunctioning (P <.001), and mental-health (P = .003) domain scores, with alb-IFN 900 mcg vs PEG-IFN. Overall, disability day assessments were significantly more favorable with albIFN 900 mcg vs PEG-IFN. The proportion of patients with ≥7 missed workdays in the month prior to assessment was significantly lower with alb-IFN 900 mcg vs PEG-IFN at wk 12 (4.2% vs 18.1%; P = .006) and 24 (5.3% vs 20.3%: P = .005). The results for albIFN 1200 mcg q2wk and q4wk were generally comparable to those for PEG IFN. Conclusions: The alb-IFN 900-mcg q2wk regimen was associated with significantly more favorable HRQOL and fewer missed workdays, while maintaining efficacy at least comparable to that of PEG-IFN.
S1934 Detection of Sen Virus in Chronic HCV Genotype 4 Infected Patients By PCR and Sequencing in Egypt Ahmed A. Monis, Samia Girgis SEN virus, was recently discovered,8 different genotypes SENV(A-H) have been described, 2 of them SENV-D and -H have been assumed to be responsible for post-transfusion hepatitis in humans. The pathogenic role of SENV in liver diseases is still controversial. Among pt's with chronic HCV infection,40% is coinfected with SENV. The effect of SENV on the course of HCV infection and response to interferon(IFN) treatment,& the effect of IFN on the clearance of SENV, are unknown. Aim:to detect SENV in healthy blood donors and in chronic HCV infected pt's, to identify the influence of SENV coinfection on clinical, virological criteria and response to combination therapy. Methods:SENV-D and -H genotypes were detected in serum from 50 pt's with chronic HCV and from 60 control subjects of healthy blood donors by PCR followed by sequencing to identify specificity. HCV Ab's, reverse transcription PCR (RT-PCR) and genotyping were detected for diagnosis. Follow up by detection of HCV viral load for all pt's under IFN and ribavirin therapy. Results:80% of chronic HCV pt's were positive for SENV DNA which was highly statistically different from the 60 blood donors who were all negative for SENV(P <0.001). By sequencing, the specificity of genotype-specific PCR was confirmed. There was a highly significant association between SENV & HCV infection (P <0.001). The pt group; 52% were infected with both types D and H,26% with SENV-H and 2% with SENV-D. The most prevalent genotype in the pt group was SENV-H(78%)followed by SENV-(54%). All patients had HCV genotype 4. A highly statistically significant was found between the pt's and control groups as regards history of blood transfusion, surgical operation,HCV-RNA and SENV-DNA(P<0.001). While no statistical significance was found between the HCV infection alone & coinfected pt's(P>0.05). Hepatosplenomegaly, anaemia and higher bilirubin were more detected among coinfected pt's with HCV & SENV (P<0.05). HCV Response rate to combination therapy did not differ significantly between SENV positive(57.5%) and negative(40%)pts(p > 0.05). In addition, HCV response occurred in 19(70%)of the 26 pts with mixed SENV-D and-H, 4(31%) in the 13 SENV-H cases and 4(40%)of the 10 only HCV infected cases. Conclusion: SENV infection is frequently detected in patients with chronic HCV and is not present in healthy blood donors. The most prevalent genotype is SENV-H & SENV-D (mainly mixed SENV-H & D). There was a significant association between SENV & HCV infection and with history of blood transfusion and surgical operation. SENV coinfection does not influence the clinical course of HCV or the response rate to combination therapy.
S1937 Type of Response to Prior Pegylated Interferon Alfa-2b (12KD)/RBV Predicts Subsequent Response to Retreatment with Peginterferon Alfa-2a (40KD)/RBV Patrick Marcellin, Bradley L. Freilich, Pietro Andreone, Adrian M. Di Bisceglie, Carlos E. Brandao-Mello, K. Rajender Reddy, Antonio Craxi, Antonio Olveira, Gerlinde Teuber, Diethelm Messinger, Greg Hooper, Matei Popescu, Donald M. Jensen Introduction: In the REPEAT study, 13-16% of non-responders to PEG-IFNα-2b/RBV achieved SVR under retreatment with 72 weeks of PEG-IFNα-2a/RBV. Patients with undetectable HCV RNA at week 12 had a high probability of achieving SVR (49%). In a retrospective analysis, we investigated if the magnitude of response to prior Peg-IFNα-2b (12KD, PegIntron®)/RBV predicted week 12 responses among patients retreated with Peg-IFNα-2a (40KD, PEGASYS®)/RBV. Methods: Patients were randomized to 4 regimens, Arms A and B PegIFNα-2a (40KD; PEGASYS®) 360µg/wk for 12 weeks then 180µg/wk for a further 60 or 36 weeks, respectively; Arms C and D Peg-IFNα-2a 180µg/wk for 72 or 48 weeks, respectively. All patients received RBV (1000/1200mg/day). For patients with evaluable HCV-RNA response at week 12 in REPEAT, we correlated quantitative HCV-RNA response between week 12 and 24 during prior Peg-IFNα-2b/RBV treatment with undetectable or unquantifiable week 12 HCV RNA in REPEAT. Responses to prior Peg-IFNα-2b were categorized as unquantifiable, ≥2 log drop, <2 log drop or unknown. Results: Of 847 patients analyzed, 310 could be categorized regarding both prior response and week 12 on-study response (108, 54, 49 and 99 for A, B, C and D, respectively). Seventeen patients achieved unquantifiable HCV-RNA with prior Peg-IFNα-2b, of these, 13 (76%) achieved week 12 undetectable/ unquantifiable HCV-RNA in REPEAT. Sixty patients achieved ≥2log drop (HCV-RNA quantifiable) with prior Peg-IFNα-2b, and of these 33 (55%) achieved week 12 undetectable/ unquantifiable HCV-RNA in REPEAT (63%, 71%, 38% and 43% for arms A, B, C and D respectively). 233 patients achieved <2log drop HCV-RNA with prior Peg-IFNα-2b and of these, 66 (28%) achieved week 12 undetectable/unquantifiable HCV-RNA in REPEAT (32%, 35%, 15%, and 27% for arms A, B, C and D respectively). 537 patients had an unknown quantitative HCV-RNA response at week 12 to 24 with prior Peg-IFNα-2b, of these, 201 (37%) achieved a week 12 undetectable/unquantifiable HCV-RNA in REPEAT. Conclusion: In non-responder patients, the type of prior response to Peg-IFNα-2b/RBV was predictive of achieving a week 12 undetectable/unquantifiable HCV-RNA with Peg-IFNα-2a/RBV retreatment, allowing physicians and patients to assess the likelihood of achieving a week 12 response with Peg-IFNα-2a and therefore the chance for SVR.
S1935 Genotype Characterization and Phylogenetic Analysis of Hepatitis B Virus in North-East Iranian Population Houshang Rafatpanah, hamid Reza Sima, Narges Valizadeh, Abulfazl Mahmood Zadeh, Narges Saed, Abbas Esmaeil Zadeh Background: Hepatitis B virus (HBV) can be classified into at least eight genotypes, A-H and four major serotypes, ayw, ayr, adw and adr, on the bases of complete genome and S gene sequence analysis. Each genotype has distinct geographic distribution. We evaluated the distribution HBV genotypes among patients with chronic infection in northeast of Iran. Methods: Seventy patients with chronic HBV infection(21 chronic hepatitis, 23 liver cirrhosis, and 26 inactive carrier) were enrolled. DNA was isolated by phenol-chloroform method. HBV genotyping was carried out by RFLP analysis (Hinf1 and Tsp509) and SSP (specific sequence primers)-PCR. The results were confirmed by sequencing. Results: and conclusion: 84.3% (69 of 70) of the patients were negative for HBeAg and 15.7% were positive for
A-787
AASLD Abstracts
AASLD Abstracts
Health-Related Quality of Life (HRQOL) with Albinterferon Alfa-2b Plus Ribavirin in IFN Treatment-NaïVe Patients with Genotype 1 Chronic Hepatitis C Stephen Pianko, Eric M. Yoshida, Stefan Zeuzem, Yves Benhamou, Vincent G. Bain, Erik Pulkstenis, John G. McHutchison, Mani Subramanian
IRES is a unique domain to mediate the viral translation. The mechanism of IRES mediated translation is not fully understood. In our study, we have designed eleven oligonucleotides according to the complementary sequences from polioviral IRES sequences. Sequence comparison showed these oligonucleotides only have complimentary sequences with poliovirus IRES, not other IRES elements. These oligonucleotides did not inhibit the translation of rabbit globin mRNA in HeLa cell extract. This data represented no inhibitory effects of oligonucleotides on cap-dependent translation. The chimeric polio viral genome containing the HCV IRES and different picornaviral IRESes sequences were used to examine the inhibitory effects of these oligonucleotides. Among these eleven oligonucleotides, one (5'CCCCCTT-3') was found to have strong inhibitory effects (>95%) on translation of type I picornaviral IRES, but not HCV IRES mediated translation. The minimal required sequence for this oligonucleotide is 5'-CCCCCTT-3'. UV cross-linking study demonstrated this oligonucleotide interacting with a 38 kd cellular protetin, poly(rC) binding protein 2 (PCBP2)(Nucleic Acids Research, 2004, Vol. 32, No. 4). Another oligonucleotide (5'-CGCGTTACG-3') showed the strongest inhibition on HCV IRES translation, but less inhibition on other IRES elements mediated translation. UV cross-linking study confirmed two cellular proteins, p68 and p70 to interact with this oligonucleotide. The identities for two cellular proteins are currently under investigation. These data showed the difference between the HCV IRES mediated translation and picornaviral IRESes mediated translation. Further investigation is necessary to explore the translational mechanism for these IRES elements as a target to develop the anti-viral chemicals.
HBeAg. HBV-DNA was positive in all samples. Detection of a 540bp PCR product indicated positive sample for HBV DNA. RFLP analysis and SSP results showed that all patients had genotype D and all isolates that were sequenced were classified as genotype D. So, results were consistent to sequencing. The phylogenetic tree was constructed using phylips software and its result showed that the isolates are derived from the same evolutionary tree. Conclusion: Genotype D is the most prevalent HBV genotype in north-east Iranian patents. PCR-RFLP can use to detect of HBV genotyping in clinical practice.
S1933 The Difference of Hepatitis C Viral (HCV) Internal Ribosomal Entry Site (Ires) and Picornaviral Ires-Mediated Translation Inhibited By Oligonucleotides Xiaoyu Li, Steffen Mueller, Eckard Wimmer
S1936
Background and Aims: This phase 2b, active-controlled study evaluated the efficacy, safety, and HRQOL of albinterferon alfa-2b (alb-IFN), a novel recombinant protein comprising IFN alfa-2b genetically fused to human albumin, in combination with oral ribavirin in IFN treatment-naïve patients with genotype 1, chronic hepatitis C (CHC). HRQOL was assessed by changes in SF-36 v2 and missed workdays due to CHC in patients treated with albIFN vs peginterferon alfa-2a (PEG-IFN). Methods: 458 patients were randomized to 4 subcutaneous treatment groups: PEG-IFN 180 mcg qwk, and alb-IFN 900 and 1200 mcg q2wk, and 1200 mcg q4wk. Primary efficacy endpoint: sustained virologic response (SVR). SF-36 was evaluated at baseline, wk 4, 12, 24, and 48 on treatment, and wk 4, 12, and 24 post-treatment. Patient disability was evaluated by number of missed workdays or days with impaired activity. Missing data were handled using the last observation carried forward. Results: By intention-to-treat analysis, SVR rates were 58.5% for alb-IFN 900 mcg, 55.5% and 50.9% for 1200 mcg q2wk and q4wk, respectively, and 57.9% for PEG-IFN (P = not significant). Overall, alb-IFN had less impact on the physical and mental domains of SF 36 during treatment vs PEG-IFN. Reductions in SF-36 scores occurred by wk 4-12, consistent with the occurrence of most treatment-associated adverse events. At wk 12, there was significantly less worsening in SF-36 physical (P = .04) and mental (P = .02) component scores, as well as in role-physical (P = .02), bodily pain (P = .003), vitality (P = .02), socialfunctioning (P <.001), and mental-health (P = .003) domain scores, with alb-IFN 900 mcg vs PEG-IFN. Overall, disability day assessments were significantly more favorable with albIFN 900 mcg vs PEG-IFN. The proportion of patients with ≥7 missed workdays in the month prior to assessment was significantly lower with alb-IFN 900 mcg vs PEG-IFN at wk 12 (4.2% vs 18.1%; P = .006) and 24 (5.3% vs 20.3%: P = .005). The results for albIFN 1200 mcg q2wk and q4wk were generally comparable to those for PEG IFN. Conclusions: The alb-IFN 900-mcg q2wk regimen was associated with significantly more favorable HRQOL and fewer missed workdays, while maintaining efficacy at least comparable to that of PEG-IFN.
S1934 Detection of Sen Virus in Chronic HCV Genotype 4 Infected Patients By PCR and Sequencing in Egypt Ahmed A. Monis, Samia Girgis SEN virus, was recently discovered,8 different genotypes SENV(A-H) have been described, 2 of them SENV-D and -H have been assumed to be responsible for post-transfusion hepatitis in humans. The pathogenic role of SENV in liver diseases is still controversial. Among pt's with chronic HCV infection,40% is coinfected with SENV. The effect of SENV on the course of HCV infection and response to interferon(IFN) treatment,& the effect of IFN on the clearance of SENV, are unknown. Aim:to detect SENV in healthy blood donors and in chronic HCV infected pt's, to identify the influence of SENV coinfection on clinical, virological criteria and response to combination therapy. Methods:SENV-D and -H genotypes were detected in serum from 50 pt's with chronic HCV and from 60 control subjects of healthy blood donors by PCR followed by sequencing to identify specificity. HCV Ab's, reverse transcription PCR (RT-PCR) and genotyping were detected for diagnosis. Follow up by detection of HCV viral load for all pt's under IFN and ribavirin therapy. Results:80% of chronic HCV pt's were positive for SENV DNA which was highly statistically different from the 60 blood donors who were all negative for SENV(P <0.001). By sequencing, the specificity of genotype-specific PCR was confirmed. There was a highly significant association between SENV & HCV infection (P <0.001). The pt group; 52% were infected with both types D and H,26% with SENV-H and 2% with SENV-D. The most prevalent genotype in the pt group was SENV-H(78%)followed by SENV-(54%). All patients had HCV genotype 4. A highly statistically significant was found between the pt's and control groups as regards history of blood transfusion, surgical operation,HCV-RNA and SENV-DNA(P<0.001). While no statistical significance was found between the HCV infection alone & coinfected pt's(P>0.05). Hepatosplenomegaly, anaemia and higher bilirubin were more detected among coinfected pt's with HCV & SENV (P<0.05). HCV Response rate to combination therapy did not differ significantly between SENV positive(57.5%) and negative(40%)pts(p > 0.05). In addition, HCV response occurred in 19(70%)of the 26 pts with mixed SENV-D and-H, 4(31%) in the 13 SENV-H cases and 4(40%)of the 10 only HCV infected cases. Conclusion: SENV infection is frequently detected in patients with chronic HCV and is not present in healthy blood donors. The most prevalent genotype is SENV-H & SENV-D (mainly mixed SENV-H & D). There was a significant association between SENV & HCV infection and with history of blood transfusion and surgical operation. SENV coinfection does not influence the clinical course of HCV or the response rate to combination therapy.
S1937 Type of Response to Prior Pegylated Interferon Alfa-2b (12KD)/RBV Predicts Subsequent Response to Retreatment with Peginterferon Alfa-2a (40KD)/RBV Patrick Marcellin, Bradley L. Freilich, Pietro Andreone, Adrian M. Di Bisceglie, Carlos E. Brandao-Mello, K. Rajender Reddy, Antonio Craxi, Antonio Olveira, Gerlinde Teuber, Diethelm Messinger, Greg Hooper, Matei Popescu, Donald M. Jensen Introduction: In the REPEAT study, 13-16% of non-responders to PEG-IFNα-2b/RBV achieved SVR under retreatment with 72 weeks of PEG-IFNα-2a/RBV. Patients with undetectable HCV RNA at week 12 had a high probability of achieving SVR (49%). In a retrospective analysis, we investigated if the magnitude of response to prior Peg-IFNα-2b (12KD, PegIntron®)/RBV predicted week 12 responses among patients retreated with Peg-IFNα-2a (40KD, PEGASYS®)/RBV. Methods: Patients were randomized to 4 regimens, Arms A and B PegIFNα-2a (40KD; PEGASYS®) 360µg/wk for 12 weeks then 180µg/wk for a further 60 or 36 weeks, respectively; Arms C and D Peg-IFNα-2a 180µg/wk for 72 or 48 weeks, respectively. All patients received RBV (1000/1200mg/day). For patients with evaluable HCV-RNA response at week 12 in REPEAT, we correlated quantitative HCV-RNA response between week 12 and 24 during prior Peg-IFNα-2b/RBV treatment with undetectable or unquantifiable week 12 HCV RNA in REPEAT. Responses to prior Peg-IFNα-2b were categorized as unquantifiable, ≥2 log drop, <2 log drop or unknown. Results: Of 847 patients analyzed, 310 could be categorized regarding both prior response and week 12 on-study response (108, 54, 49 and 99 for A, B, C and D, respectively). Seventeen patients achieved unquantifiable HCV-RNA with prior Peg-IFNα-2b, of these, 13 (76%) achieved week 12 undetectable/ unquantifiable HCV-RNA in REPEAT. Sixty patients achieved ≥2log drop (HCV-RNA quantifiable) with prior Peg-IFNα-2b, and of these 33 (55%) achieved week 12 undetectable/ unquantifiable HCV-RNA in REPEAT (63%, 71%, 38% and 43% for arms A, B, C and D respectively). 233 patients achieved <2log drop HCV-RNA with prior Peg-IFNα-2b and of these, 66 (28%) achieved week 12 undetectable/unquantifiable HCV-RNA in REPEAT (32%, 35%, 15%, and 27% for arms A, B, C and D respectively). 537 patients had an unknown quantitative HCV-RNA response at week 12 to 24 with prior Peg-IFNα-2b, of these, 201 (37%) achieved a week 12 undetectable/unquantifiable HCV-RNA in REPEAT. Conclusion: In non-responder patients, the type of prior response to Peg-IFNα-2b/RBV was predictive of achieving a week 12 undetectable/unquantifiable HCV-RNA with Peg-IFNα-2a/RBV retreatment, allowing physicians and patients to assess the likelihood of achieving a week 12 response with Peg-IFNα-2a and therefore the chance for SVR.
S1935 Genotype Characterization and Phylogenetic Analysis of Hepatitis B Virus in North-East Iranian Population Houshang Rafatpanah, hamid Reza Sima, Narges Valizadeh, Abulfazl Mahmood Zadeh, Narges Saed, Abbas Esmaeil Zadeh Background: Hepatitis B virus (HBV) can be classified into at least eight genotypes, A-H and four major serotypes, ayw, ayr, adw and adr, on the bases of complete genome and S gene sequence analysis. Each genotype has distinct geographic distribution. We evaluated the distribution HBV genotypes among patients with chronic infection in northeast of Iran. Methods: Seventy patients with chronic HBV infection(21 chronic hepatitis, 23 liver cirrhosis, and 26 inactive carrier) were enrolled. DNA was isolated by phenol-chloroform method. HBV genotyping was carried out by RFLP analysis (Hinf1 and Tsp509) and SSP (specific sequence primers)-PCR. The results were confirmed by sequencing. Results: and conclusion: 84.3% (69 of 70) of the patients were negative for HBeAg and 15.7% were positive for
A-787
AASLD Abstracts
AASLD Abstracts
Health-Related Quality of Life (HRQOL) with Albinterferon Alfa-2b Plus Ribavirin in IFN Treatment-NaïVe Patients with Genotype 1 Chronic Hepatitis C Stephen Pianko, Eric M. Yoshida, Stefan Zeuzem, Yves Benhamou, Vincent G. Bain, Erik Pulkstenis, John G. McHutchison, Mani Subramanian
IRES is a unique domain to mediate the viral translation. The mechanism of IRES mediated translation is not fully understood. In our study, we have designed eleven oligonucleotides according to the complementary sequences from polioviral IRES sequences. Sequence comparison showed these oligonucleotides only have complimentary sequences with poliovirus IRES, not other IRES elements. These oligonucleotides did not inhibit the translation of rabbit globin mRNA in HeLa cell extract. This data represented no inhibitory effects of oligonucleotides on cap-dependent translation. The chimeric polio viral genome containing the HCV IRES and different picornaviral IRESes sequences were used to examine the inhibitory effects of these oligonucleotides. Among these eleven oligonucleotides, one (5'CCCCCTT-3') was found to have strong inhibitory effects (>95%) on translation of type I picornaviral IRES, but not HCV IRES mediated translation. The minimal required sequence for this oligonucleotide is 5'-CCCCCTT-3'. UV cross-linking study demonstrated this oligonucleotide interacting with a 38 kd cellular protetin, poly(rC) binding protein 2 (PCBP2)(Nucleic Acids Research, 2004, Vol. 32, No. 4). Another oligonucleotide (5'-CGCGTTACG-3') showed the strongest inhibition on HCV IRES translation, but less inhibition on other IRES elements mediated translation. UV cross-linking study confirmed two cellular proteins, p68 and p70 to interact with this oligonucleotide. The identities for two cellular proteins are currently under investigation. These data showed the difference between the HCV IRES mediated translation and picornaviral IRESes mediated translation. Further investigation is necessary to explore the translational mechanism for these IRES elements as a target to develop the anti-viral chemicals.