- Email: [email protected]
HCV coinfection
146
Poster Sessions
gens did not change significantly with treatment, independently of HCV genotype. As a further control of these negative results obtained with a limited set of peptides and tetramers, 4 patients were analyzed by Elispot with a panel of 600 overlapping 15-mer peptides covering the whole HCV sequence. No significant difference in Elispot responses was observed following treatment. Our results indicate that a rapid second phase viral decline cannot be explained by a strong systemic antigen-specific T-cell response. Other immune or non-immune correlates should be identified.
495 STEATOSIS AND FIBROSIS IN CHRONIC HEPATITIS C: RELATIVE ROLE OF LIPID PEROXIDATION, LEPTINEMIA AND INSULIN RESISTANCE
A. Muzzi 1 , G. Leandro 2 , L. Rubbia-Brandt 1 , R. James 1 , O. Keiser 3 , R. Malinverni 4 , J.F. Dufour 5 , B. Helbling 6 , A. Hadengue 1 , J.J. Gonvers 3 , B. Mullhaupt 7 , A. Cerny 8 , M. Mondelli 9 , F. Negro 1 . The Swiss Hepatitis C Cohort Study; 1 University of Geneva, Geneva, Switzerland; 2 IRCCS De Bellis, Castellana Grotte, Italy; 3 Chuv, Lausanne, Switzerland; 4 Hospital De Cadolle, Neuchatel, Switzerland; 5 University Hospital, Berne, Switzerland; 6 Cantonal Hospital, St. Gallen, Switzerland; 7 University Hospital, Zurich, Switzerland; 8 City Hospital, Lugano, Switzerland; 9 University of Pavia, Pavia, Switzerland Liver steatosis is a frequent finding in chronic hepatitis C. An association has been suggested between steatosis and fibrosis progression rate, but the pathogenetic mechanisms linking fatty infiltration and collagen deposition are unknown. We assessed the levels of lipid peroxidation (as serum TBArs), leptinemia and insulin resistance (as HOMA score) in 243 chronic hepatitis C patients consecutively recruited at 7 clinical centers across Switzerland. These and several other clinical, virological and histological parameters were analysed in a multivariable logistic regression aimed at identifying independent factors associated with liver steatosis and fibrosis. When all patients were considered, steatosis was associated with genotype 3 (P<0.001), alcohol abuse (P=0.001), serum HCV RNA (P=0.002) and presence of fibrosis (P=0.026). Fibrosis was independently predicted by inflammatory activity (P<0.001), age (P<0.001) and serum level of lipid peroxidation end-products (P=0.01). In patients with genotype 3 (n=76), steatosis was strongly associated with fibrosis (P<0.001) and HCV RNA level (P=0.003), whereas in patients with genotype non-3 (n=167) the association held only for ongoing alcohol abuse (P<0.001). Fibrosis was independently associated with inflammation (P=0.003), age (P=0.017) and presence of steatosis (P=0.015) in patients with genotype 3, while in those with genotype non-3 infection the association was shown with inflammation (P=0.001), age (P<0.001) and lipid peroxidation levels (P=0.024). In conclusion, although lipid peroxidation may be linked to more rapid liver fibrosis progression, it does not seem to be influenced by the presence of liver steatosis. Neither leptin nor insulin resistance appear to play a role in fibrogenesis in chronic hepatitis C.
496 INCREASED INTRAHEPATIC CYCLOOXYGENASE-2 AND METALLOPROTEINASE-2 AND -9 EXPRESSION ASSOCIATES WITH ADVANCED LIVER DISEASE IN CHRONIC HEPATITIS C VIRUS INFECTION: ROLE OF VIRAL CORE AND NS5A PROTEINS
O. Nunez 1 , A. Fernandez-Martinez 2 , A. Apolinario 1 , R. Lorente 1 , L. Bosca 2 , G. de la Cruz 1 , G. Clemente 1 , P. Martin-Sanz 2 , C. Garcia-Monzon 1 . 1 Instituto De Hepatologia Clinica-Experimental Y Trasplante Hepatico, Hospital Universitario Gregorio Maranon, Santa Cristina, Spain; 2 Instituto De Bioquimica (CSIC-UCM) Facultad De Farmacia, Universidad Complutense, Centro Nacional De Enfermedades Cardiovasculares-CNIC*, Madrid, Spain Background & Aims: We aimed to determine the expression pattern of COX-2, MMP-2 and -9 in hepatitis C virus (HCV) infection.
Methods: COX-2, MMP-2, and MMP-9 expression was assessed by Western blot, RT-PCR, and enzyme activity in HCV liver biopsies (mild chronic hepatitis (MCH)=22, severe chronic hepatitis (SCH)=5, end-stage cirrhosis (CIR)=5), and from 10 non-viral liver samples. COX-2 and MMP-9 expression was assessed by Western blot and enzyme activity in Chang Liver (CHL) cells stably transfected with core and NS5A viral proteins. Results: COX-2 protein and mRNA levels were significantly higher in MCH (2.4 and 2.6-fold, respectively), SCH (3.3 and 3.5-fold) and in CIR (3.9 and 5.3-fold) than in control samples. A progressive increase in PGE2 levels was found in agreement with COX-2 expression. There was a statistically significant increase of MMP-2 and MMP-9 proteins in MCH (3.2 and 2-fold, respectively), SCH (4.7 and 2.6-fold) and CIR (3.8 and 6.6-fold) as well as MMP-9 activity in SCH (2-fold) and CIR (2.8-fold), as compared with controls. We further observed that COX-2 protein and activity was induced in resting and activated CHL cells transfected with core and NS5A proteins. An inducer effect on MMP-9 synthesis in CHL cells was exerted by core but not NS5A, being this induction partially abrogated by a specific COX-2 inhibitor. Conclusions: The intrahepatic expression level of COX-2 and MMP-2 and -9 significantly correlated with the histological severity of HCV-related chronic liver disease. A virus-induced hepatocellular COX-2 and MMP-9 upregulation, partially abrogated by COX-2 inhibition, was clearly shown.
497 AN OPEN-LABEL, MULTICENTER STUDY OF THE EFFICACY AND SAFETY OF PEGINTERFERON ALFA-2A (40KD) (PEGASYS ), ALONE OR IN COMBINATION WITH RIBAVIRIN (COPEGUS ), IN PATIENTS WITH HIV/HCV COINFECTION
E. Ortega 1 , A. Martin 1 , C. Barros 2 , K. Aguirrebengoa 3 , J. Gonzalez 4 , A. Garcia 5 , R. Rubio 6 , F. Pulido 6 . 1 Hospital General Universitario, Valencia, Spain; 2 Hospital De Mostoles, Madrid, Spain; 3 Hospital De Cruces, Bilbao, Spain; 4 Hospital La Paz, Madrid, Spain; 5 Hospital Santa Maria Del Rosell, Cartagena, Spain; 6 Hospital 12 De Octubre, Madrid, Spain Background: Liver disease has emerged as a major cause of morbidity and mortality in patients with HIV/HVC coinfection, mainly as a result of coinfection or hepatotoxicity induced by antiretroviral drugs. Objective: To evaluate the efficacy and safety of peginterferon alfa-2a treatment alone or with ribavirin. Methods: Patients initially received peginterferon alfa-2a 180 mcg/week alone. HCV-RNA levels were monitored to assess virological response (VR). At week 4, patients with VR (<50 IU/mL HCV RNA) remained on peginterferon alfa-2a alone while those with continual viremia (≥50 IU/mL HCV RNA) had ribavirin 800 mg/day + peginterferon alfa-2a. This study is ongoing. Results: 311 patients have been recruited (227 males/82 females, mean age 38 years, 47% HCV genotype 1, mean ALT 118 U/L, 30% have bridging fibrosis or cirrhosis, mean CD4+ cells counts are 647/mm3, 82% are on HAART). 298 patients completed 4 weeks of therapy, of which 24% have had an early VR. 49% (129/260) and 65% (141/216) of patients had a VR at weeks 12 and 24, respectively. At the moment, data of week 48 is available for 104 patients and 94 of them have VR. Complete data of EOT and EOF will be available for March 2004. Both treatment regimens were generally well tolerated. Discontinuation was observed in 76 patients (25,5%), 38 of these withdrawals were due to adverse events (related o not to treatment), among those, 13 were laboratory abnormalities. Conclusion: Peginterferon alfa-2a alone or in combination with ribavirin seems to be an effective and safe treatment in patients coinfected.
Poster Sessions
gens did not change significantly with treatment, independently of HCV genotype. As a further control of these negative results obtained with a limited set of peptides and tetramers, 4 patients were analyzed by Elispot with a panel of 600 overlapping 15-mer peptides covering the whole HCV sequence. No significant difference in Elispot responses was observed following treatment. Our results indicate that a rapid second phase viral decline cannot be explained by a strong systemic antigen-specific T-cell response. Other immune or non-immune correlates should be identified.
495 STEATOSIS AND FIBROSIS IN CHRONIC HEPATITIS C: RELATIVE ROLE OF LIPID PEROXIDATION, LEPTINEMIA AND INSULIN RESISTANCE
A. Muzzi 1 , G. Leandro 2 , L. Rubbia-Brandt 1 , R. James 1 , O. Keiser 3 , R. Malinverni 4 , J.F. Dufour 5 , B. Helbling 6 , A. Hadengue 1 , J.J. Gonvers 3 , B. Mullhaupt 7 , A. Cerny 8 , M. Mondelli 9 , F. Negro 1 . The Swiss Hepatitis C Cohort Study; 1 University of Geneva, Geneva, Switzerland; 2 IRCCS De Bellis, Castellana Grotte, Italy; 3 Chuv, Lausanne, Switzerland; 4 Hospital De Cadolle, Neuchatel, Switzerland; 5 University Hospital, Berne, Switzerland; 6 Cantonal Hospital, St. Gallen, Switzerland; 7 University Hospital, Zurich, Switzerland; 8 City Hospital, Lugano, Switzerland; 9 University of Pavia, Pavia, Switzerland Liver steatosis is a frequent finding in chronic hepatitis C. An association has been suggested between steatosis and fibrosis progression rate, but the pathogenetic mechanisms linking fatty infiltration and collagen deposition are unknown. We assessed the levels of lipid peroxidation (as serum TBArs), leptinemia and insulin resistance (as HOMA score) in 243 chronic hepatitis C patients consecutively recruited at 7 clinical centers across Switzerland. These and several other clinical, virological and histological parameters were analysed in a multivariable logistic regression aimed at identifying independent factors associated with liver steatosis and fibrosis. When all patients were considered, steatosis was associated with genotype 3 (P<0.001), alcohol abuse (P=0.001), serum HCV RNA (P=0.002) and presence of fibrosis (P=0.026). Fibrosis was independently predicted by inflammatory activity (P<0.001), age (P<0.001) and serum level of lipid peroxidation end-products (P=0.01). In patients with genotype 3 (n=76), steatosis was strongly associated with fibrosis (P<0.001) and HCV RNA level (P=0.003), whereas in patients with genotype non-3 (n=167) the association held only for ongoing alcohol abuse (P<0.001). Fibrosis was independently associated with inflammation (P=0.003), age (P=0.017) and presence of steatosis (P=0.015) in patients with genotype 3, while in those with genotype non-3 infection the association was shown with inflammation (P=0.001), age (P<0.001) and lipid peroxidation levels (P=0.024). In conclusion, although lipid peroxidation may be linked to more rapid liver fibrosis progression, it does not seem to be influenced by the presence of liver steatosis. Neither leptin nor insulin resistance appear to play a role in fibrogenesis in chronic hepatitis C.
496 INCREASED INTRAHEPATIC CYCLOOXYGENASE-2 AND METALLOPROTEINASE-2 AND -9 EXPRESSION ASSOCIATES WITH ADVANCED LIVER DISEASE IN CHRONIC HEPATITIS C VIRUS INFECTION: ROLE OF VIRAL CORE AND NS5A PROTEINS
O. Nunez 1 , A. Fernandez-Martinez 2 , A. Apolinario 1 , R. Lorente 1 , L. Bosca 2 , G. de la Cruz 1 , G. Clemente 1 , P. Martin-Sanz 2 , C. Garcia-Monzon 1 . 1 Instituto De Hepatologia Clinica-Experimental Y Trasplante Hepatico, Hospital Universitario Gregorio Maranon, Santa Cristina, Spain; 2 Instituto De Bioquimica (CSIC-UCM) Facultad De Farmacia, Universidad Complutense, Centro Nacional De Enfermedades Cardiovasculares-CNIC*, Madrid, Spain Background & Aims: We aimed to determine the expression pattern of COX-2, MMP-2 and -9 in hepatitis C virus (HCV) infection.
Methods: COX-2, MMP-2, and MMP-9 expression was assessed by Western blot, RT-PCR, and enzyme activity in HCV liver biopsies (mild chronic hepatitis (MCH)=22, severe chronic hepatitis (SCH)=5, end-stage cirrhosis (CIR)=5), and from 10 non-viral liver samples. COX-2 and MMP-9 expression was assessed by Western blot and enzyme activity in Chang Liver (CHL) cells stably transfected with core and NS5A viral proteins. Results: COX-2 protein and mRNA levels were significantly higher in MCH (2.4 and 2.6-fold, respectively), SCH (3.3 and 3.5-fold) and in CIR (3.9 and 5.3-fold) than in control samples. A progressive increase in PGE2 levels was found in agreement with COX-2 expression. There was a statistically significant increase of MMP-2 and MMP-9 proteins in MCH (3.2 and 2-fold, respectively), SCH (4.7 and 2.6-fold) and CIR (3.8 and 6.6-fold) as well as MMP-9 activity in SCH (2-fold) and CIR (2.8-fold), as compared with controls. We further observed that COX-2 protein and activity was induced in resting and activated CHL cells transfected with core and NS5A proteins. An inducer effect on MMP-9 synthesis in CHL cells was exerted by core but not NS5A, being this induction partially abrogated by a specific COX-2 inhibitor. Conclusions: The intrahepatic expression level of COX-2 and MMP-2 and -9 significantly correlated with the histological severity of HCV-related chronic liver disease. A virus-induced hepatocellular COX-2 and MMP-9 upregulation, partially abrogated by COX-2 inhibition, was clearly shown.
497 AN OPEN-LABEL, MULTICENTER STUDY OF THE EFFICACY AND SAFETY OF PEGINTERFERON ALFA-2A (40KD) (PEGASYS ), ALONE OR IN COMBINATION WITH RIBAVIRIN (COPEGUS ), IN PATIENTS WITH HIV/HCV COINFECTION
E. Ortega 1 , A. Martin 1 , C. Barros 2 , K. Aguirrebengoa 3 , J. Gonzalez 4 , A. Garcia 5 , R. Rubio 6 , F. Pulido 6 . 1 Hospital General Universitario, Valencia, Spain; 2 Hospital De Mostoles, Madrid, Spain; 3 Hospital De Cruces, Bilbao, Spain; 4 Hospital La Paz, Madrid, Spain; 5 Hospital Santa Maria Del Rosell, Cartagena, Spain; 6 Hospital 12 De Octubre, Madrid, Spain Background: Liver disease has emerged as a major cause of morbidity and mortality in patients with HIV/HVC coinfection, mainly as a result of coinfection or hepatotoxicity induced by antiretroviral drugs. Objective: To evaluate the efficacy and safety of peginterferon alfa-2a treatment alone or with ribavirin. Methods: Patients initially received peginterferon alfa-2a 180 mcg/week alone. HCV-RNA levels were monitored to assess virological response (VR). At week 4, patients with VR (<50 IU/mL HCV RNA) remained on peginterferon alfa-2a alone while those with continual viremia (≥50 IU/mL HCV RNA) had ribavirin 800 mg/day + peginterferon alfa-2a. This study is ongoing. Results: 311 patients have been recruited (227 males/82 females, mean age 38 years, 47% HCV genotype 1, mean ALT 118 U/L, 30% have bridging fibrosis or cirrhosis, mean CD4+ cells counts are 647/mm3, 82% are on HAART). 298 patients completed 4 weeks of therapy, of which 24% have had an early VR. 49% (129/260) and 65% (141/216) of patients had a VR at weeks 12 and 24, respectively. At the moment, data of week 48 is available for 104 patients and 94 of them have VR. Complete data of EOT and EOF will be available for March 2004. Both treatment regimens were generally well tolerated. Discontinuation was observed in 76 patients (25,5%), 38 of these withdrawals were due to adverse events (related o not to treatment), among those, 13 were laboratory abnormalities. Conclusion: Peginterferon alfa-2a alone or in combination with ribavirin seems to be an effective and safe treatment in patients coinfected.