- Email: [email protected]
Genotype mixtures of hepatitis B virus in patients treated with interferon
Poster Sessions
116
Conclusion: 1. Treatment with IFN alpha did not improve histological fibrosis in children with chronic hepatitis B; 2. However, IFN decreased surrogate serum markers of fibrogenesis (laminin-2, collagen IV, MMP-2) on the long term, while serum MMP-9/TIMP-1 which appears to mirror fibrolysis was increased; 3. The evolution of serum fibrosis markers may be more sensitive to detect slight antifibrotic effects than semiquantitative follow-up histology.
~-]
EVALUATIONOF LIVER FIBROSIS BY COMBINED SERUM MARKERS IN CHRONIC HEPATITIS C PATIENTS TREATED BY INTERFERON ALPHA AND RIBAVRIN
Vincent Leroy I , Candice Trocme 2, Serge Bottari 3, Nathalie Sturm 4, Francoise Morel 2, Jean-Pierre Zarski I . 1Departmentof
Hepatogastroenterology, CHU Grenoble; 2Laboratory of Enzymology, CHU Grenoble; 3Laboratory of biochemistry, CHU Grenoble; 4Laboratory of Cellular Pathology, CHU Grenoble, France Our aims were to evaluate the diagnostic ability of a panel of serum markers to stage liver fibrosis and to determine their kinetic during antiviral therapy. Methods: Among 194 naive patients with liver biopsy, 76 were treated by IFN and ribavirin for at least 24 weeks. The levels of MMP-1, 2 and 9, TIMP-1 and 2, PIIINP (ELISA) and HA (RIA) were measured from serum collected the day of the biopsy and every 3 months during the treatment. METAVIR score was used. Results: MMP-2 (r = 0.28; p < 0.01), TIMP- 1 (r = 0.42; p < 0.001), HA (r = 0.50; p < 0.001) and PIIINP (r = 0.62; p < 0.0001) were positively correlated with Metavir fibrosis, whereas MMP-1 (r = -0.34; r < 0.001) and MMP-2 (r = -0.24; r < 0.01) were negatively correlated. By multivariate analysis, PIIINP and MMP-1 were independently associated with fibrosis. A score calculated by combining these two variables (logistic regression function) was able to discriminate F0/F1 from F2/F3/F4 with a sensitivity of 60% and a specificity of 91% (ROC curves). During treatment, MMP-1 significantly increase and PIIINP as well as combined score significantly decrease in virological responders but not in non responders. Conclusion: Non invasive markers might be useful for estimating liver fibrosis. IFN and ribavirin reduce liver fibrosis in virological responders by decreasing fibrogenesis as well as increasing fibrolysis.
GRADING OF HEPATITIS C RELATED LIVER • TNON-INVASIVE • DISEASE: MICROBUBBLE-ENI'IANCED ULTRASOUND STUDIES Adrian Liml'2, Nayna Patel 1.2, Gavin Hamilton L2, Martin Blomiey 1, David Cosgrove 1, Graham Foster 2, Robert Goldin 3, Howard Thomas 2, Simon Taylor-Robinson 1.2.1Department of Imaging, Imperial College,
(4.9) respectively (Kruskal Wallis, p < 0.001). An AT < 20 s and CDT < 10 s was 100% sensitive for cirrhosis but only 69% and 72% specific respectively, as subjects with fibrosis sometimes showed early AT and CDT. 10 interferon-treated patients showed earlier arrival times than comparative untreated subjects. Conclusion: AT and CDT measurements hold promise in charaeterising HCV liver disease and are highly sensitive markers of cirrhosis. Treatment with interferon appears to prolong AT. This is important for disease monitoring and may be useful in assessing the efficacy of treatment regimes non-invasively and possibly replace repeat liver biopsy in some situations.
• TLOW 3RISK"OF]VERTICAL HEPATITIS C VIRUS (HCV) INFECTION IN A LARGE COHORT OF HUMAN IMMUNODEFICIENCY VIRUS (HIV)-SERONEGATIVE PREGNANT WOMEN Marina Losa l, Alicia Saez 2, Oreste Lo Iacono 3, Carlos Lozano 2, Carmelo Garcia-Monzon 1. l Hepatology Unit, UniversityHospital Santa
Cristina, Madrid; 2Serologyand Biochemistry Service, University Hospital Santa Cristina, Madrid, Spain; JGastmentemlogy Department, University of Palermo, Italy Given that conflicting results are currently available on vertical HCV transmission, the aims of this study were to 'determine the prevalence of chronic HCV infection in a large cohort of HIV-negative pregnant women and to analyze the rate of vertical HCV transmission, assessing the possible related risk factors. We prospectively recruited 20,612 consecutive unselected pregnant women who were referred to the Obstetric Unit of the University Hospital Santa Cristina. HCV-RNA was assessed in blood samples from anti-HCV+ mothers at first and third trimester of pregnancy and from their newborns at birth and after 6, 12, 24, and 36 months. Viral load and genotype were determined to all viremic women at the time of delivery as well as to viremic infants. Antibodies to HCV was detected in 119 mothers (0.57%), whereas HCV-RNA tested positive in 67% of anti-HCV+ women (n = 80). The median viral load of these viremic women, at time of delivery, was 335,300 copies per milliliter (range: 11,000-2,625,200), and the vast majority of them had a low viral load. HCV genotyping revealed subtype 1 in 60 cases (75%). Only 2 newborns tested positive for HCV-RNA immediately after birth (transmission rate: 2.4%), remaining positive during the entire follow-up. These vertically-infected babies had the same HCV genotype (lb) that their respective mothers. In conclusion, vertical HCV transmission is an infrequent event among HIV-negative HCV-infected women and, in the present study, no apparent risk factor influencing the transmission rate was identified.
~-~
GENOTYPE MIXTURES OF HEPATITIS B VIRUS IN PATIENTS TREATED WITH INTERFERON
Hammersmith Hospital, Du Cane Road, London; 2Departmentof Medicine, Imperial College, St. Mary's Hospital, Praed Street, London; 3Department of Histopathology, Imperial College, St. Mary's Hospital, Praed Street, London, UK
Charles Hannoun 1, Magnus Lindh 1, Kim Krogsgaard 2. 1Dept. of Clinical
Purpose: Non-invasive assessment of the severity of liver disease is difficult. We evaluated hepatic vein arrival times (AT) of an ultrasound microbubble agent and carotid-delay times (CDT) in grading liver disease in patients with chronic hepatitis C (HCV) infection. Materials/Method: 61 patients were studied. Grading of fibrosis (F) and inflammatory activity (NI) was carded out using the modified HA1 (Ishak) system. Patients were divided into mild hepatitis, (F < 3/6, NI < 4/18); moderate/severe hepatitis, (F > 2/6, NI > 3/18) and cirrhosis, (F = 6). Timeintensity curves of hepatic vein spectral Doppler signals and audio intensity from the carotid artery were analysed after an intravenous microbubble bolus. CDT was calculated as the difference between carotid and hepatic vein arrival times. Results: There was a monotonic decrease in the mean ATs (1 s.d.) and CDTs (1 s.d.) for mild, moderate/severe hepatitis and cirrhosis: 50.5 s (24.2), 33.6 s (26.1), 14.8 s (4.5) and 36.8 s (29), 18.7 s (22.6) and 5.8 s
kground: Little is known about co-infection with several hepatitis B virus genotypes, although previous reports of recombination and genotype shifts indicate that this should occur. Aim: To evaluate genotypic co-infection in HBV infection. Method: Using a genotypic core region PCR that identifies mixtures even if one genotype predominates, serum samples from before and after interferon treatment of 30 HBeAg positive patients were analysed. Results: Co-infection with genotype A and another genotype was found in 19 (63%) of 30 HBeAg positive patients treated with interferon. In 7 patients co-infection or genotype shifts were detectable also by direct sequencing or standard preS genotyping. In most cases, genotype changes were detected after a more than 2 log decrease or increase of the HBV DNA level. The presence of genotype mixtures did not significantly influence IFN response, but this has to be further analysed.
Virology, Goteborg University, Goteborg, Sweden; 2Clinical Research Unit, Hvidovre Hospital, Copenhagen, Denmark
Category 6: Viral hepatitis: clinical aspects
Condusion: Genotypic co-infection appears to be frequent. The finding may be relevant for tracing source of HBV infection and possibly for treatment.
[-~
MUTATIONS IN X REGION HAVE LITTLE IMPACT ON RESPONSE TO INTERFERON THERAPY FOR CHRONIC HEPATITIS B
Charles Hannoun i, Kim Krogsgaard 2, Peter Horal 1, Magnus Lindh 1.
1Department of Clinical Virology, Gtteborg University, Gtteborg, Sweden; 2Clinical Research Unit, Hvidovre Hospital, Copenhagen, Denmark
117
+ RBV 1-1.2 g/qd + AMA 200 mg/qd Group B: IFN alpha-2a 3 MU tiw sc + RBV 1-1.2 g/qd + AMA 200 mg/qd Group 3: IFN alpha-2a 3 MU tiw sc + RBV 1-1.2 g/qd R e s d t s : Of 268 so far enrolled pts, 55.5% were male, mean age was 51 yrs (range 20-65): genotype 1 was found in 57.6% of cases. Severe fibrosis or cirrhosis were found in 23.3%. The treatments were well tolerated; no major safety concerns were noted at the moment. One hundred and thirty pts have completed 3 mos therapy: at this time the percentages of pts with normal ALT levels were 51% in group A, 55% in group B, and 51% in group C. The rate of virological response was 54%, 49% and 42%, respectively. This study is ongoing and more data are needed to prove or disprove the value of triple therapy over the current standard therapy.
Background: Therapy for chronic hepatitis B with interferon-alpha (IFN) may result in viral clearance and hepatitis B e seroconversion in 30--40% of patients. It is still unclear if viral genetic variability influences response rates. However, certain core promoter mutations were recently associated with a better response to interferon. Aim" To analyse the importance of mutations in the core promoter for IFN response. Method: The entire X region, including the core promoter, of hepatitis B virus (HBV) from 26 HBeAg positive patients treated with IFN for 12 weeks, was sequenced. Serum samples from before treatment, at end-oftreatment and at follow-up of sustained and non-sustained responders were analysed. Most patients were of European origin and showed moderate aminotransferase elevation (mean 2.4 x upper limit of normal) and genotype A infection. Results: Before treatment, 21 patients had an X region identical with a consensus sequence of the corresponding genotype, and in the remaining 9 patients a mean of 1.3 mutations were found. After treatment, 1-4 new mutations (mean 1.8) had emerged in 5 patients. There was no association between specific mutations, or the number of mutations, and response to IFN. Conclusion: Analysis of X region/core promoter is of limited clinical value in HBeAg positive patients and emerging mutations in this region are not major determinants of IFN response.
I•-'•
A 3-ARM RANDOMIZED STUDY EVALUATING THE EFFICACY OFTRIPLE THERAPY OF PEGYLATED OR RECOMBINANT INTERFERON alpha-2a 3 MU BOTH COMBINED WITH RIBAVlRIN AND AMANTADINE VERSUS DUAL THERAPY OF RECOMBINANT INTERFERON alpha-2a PLUS RIBAVlRIN IN NAIVE PATIENTS WITH CHRONIC HCV HEPATITIS
Alessandra Mangia I , Giovanni Ricci2, Nicola Minerva 3, Marcello Persico 4, Vito Carretta 5, Donato Bacca 6, Franco Vinelli 7, Giuseppe Sogafi s, Vincenzo Guadagnino 9, Giovanni Maio, Angelo Andriulli 1. t Gastroenterology Division Hospital 'CSS' San
Giovanni Rotondo; 2Internal Medicine University 'La Sapienza' Roma; 3Medicine Division Hospital Canosa; 4Medicine Division Hospital 'Cotugno' Napoli; 5Medicine Division Hospital Venosa; 6Medicine Division Hospital Casarano; 7Gastroenterology Division Ospedali Riuniti Foggia," SMedicine Division Hospital 'SS.Annunziata' Taranto; 91nternal Medicine Division University Catanzaro Infectious Disease Division Hospital 'Rummo' Benevento, Italy
Background: We have previously shown that the addition of amantadine (AMA) to recombinant IFN alpha-2a was able to increase the efficacy of IFN monotherapy in pts with chronic hepatitis C (CHC). No data are available on naive pts to document whether triple therapy of AMA + ribavirin (RBV) and pegylated IFN (PEG 1FN) (group A) or recombinant IFN alpha-2a (IFN) (group B) improves the success rate over dual therapy with recombinant IFN + RIBA. Methods: 354 naive pts with serologically and biopy-proven CHC, persistently, elevated ALT and positive HCV RNA were randomized to a 3-arrn multicentre trial, as follows: Group A: PEG IFN alpha-2a 180 mcg sc qw
[-~
IL-10 ATA HAPLOTYPE AND 1ST INTERON IFN-GAMMA POLYMORPHISMS ARE MORE FREQUENT IN PTS WITH SPONTANEOUS HCV CLEARANCE
A. Mangia, R. Santoro, O. D'Apolito, A. Lacobellis, A. Andruilli.
1Gastroenterology IRCCS 'Casa Sollievo Della Sofferenza', San Giovanni Rotondo, Italy In chronic HCV infection, cytokines and their receptors play a critical role in regulating HCV immune response. The individual capacity for cytokine secretion has a major genetic component. Three different haplotype of IL-10 promoter region are associated with different serum levels of this cytokine, ATA haplotype correlating with low serum levels. Aim: to investigate the association between allelic polymorphisms of IFNy, IL-10, TNFc~, and spontaneous or IFN-induced (LTR) HCV RNA clearance. Methods: -1117, -854, -627 single nucleotide polymorphism (SNP) of IL-10 promoter by SSP and - 3 0 8 and - 2 3 8 SNP of TNFot by PCR and restriction enzymes digestion were evaluated. Microsatellite analysis of (ca) n at the 1st intron of IFNy was evaluated by PCR/acrylamide gel. Patient: 149 HCVRNA +ve pts: 105 with abnormal and 44 with normal ALT, and 78 HCV RNA - v e , 39 with past acute hepatitis and 39 long term responders to antivirals were evaluated. As controls 150 subjects from general population were assessed. Results are shown in the table HCV R N A + v e ALT> ALT N IL-10 ATA 3.3%* IFNyI25 bp 3.3%** TNFa -308AA 1% -238AA 1% *p = 0.07; ** p = 0.01
Spontaneous RNA clearance
LTR
Controls
6.6%
10.5%* 10.2%**
7.5% 3.8%
4.8% 4.8%
0 0
0 0
0 0
0 0
In conclusion: spontaneous HCV clearance is associated with ATA haplotype of IL-10 and polymorphisms of the 1st intron of IFNy.
~']
STUDY OF SEROLOGICAL MARKERS OF HBV IN POSTRANSFUSIONAL HEPATITIS C
Mafia Dolores Martin Arranz 1, Santos Del Campo Terron 2, Javier Moreno Garcia 2, Rafael Barcena Marugan 2. 1University Hospital La Paz.
Madrid; 2University Hospital Ramon y Cajal. Madrid, Spain Objetive: The aim of this study was to assess the influence of past virus B infection in the development of postransfusional chronic hepatitis C (HCV). Patients and Methods: We enrolled 184 patients with postransfusional HCV (previous transfusion without other causes of hepatitis), older than 18 years, liver biopsy at least 4 years after transfusion. None of them had HBsAg. All sera were collected at the time of liver biopsy. HBsAg, HBcAc, HBsAc were assessed by ELISA (Abbott). Serum and liver tissue
116
Conclusion: 1. Treatment with IFN alpha did not improve histological fibrosis in children with chronic hepatitis B; 2. However, IFN decreased surrogate serum markers of fibrogenesis (laminin-2, collagen IV, MMP-2) on the long term, while serum MMP-9/TIMP-1 which appears to mirror fibrolysis was increased; 3. The evolution of serum fibrosis markers may be more sensitive to detect slight antifibrotic effects than semiquantitative follow-up histology.
~-]
EVALUATIONOF LIVER FIBROSIS BY COMBINED SERUM MARKERS IN CHRONIC HEPATITIS C PATIENTS TREATED BY INTERFERON ALPHA AND RIBAVRIN
Vincent Leroy I , Candice Trocme 2, Serge Bottari 3, Nathalie Sturm 4, Francoise Morel 2, Jean-Pierre Zarski I . 1Departmentof
Hepatogastroenterology, CHU Grenoble; 2Laboratory of Enzymology, CHU Grenoble; 3Laboratory of biochemistry, CHU Grenoble; 4Laboratory of Cellular Pathology, CHU Grenoble, France Our aims were to evaluate the diagnostic ability of a panel of serum markers to stage liver fibrosis and to determine their kinetic during antiviral therapy. Methods: Among 194 naive patients with liver biopsy, 76 were treated by IFN and ribavirin for at least 24 weeks. The levels of MMP-1, 2 and 9, TIMP-1 and 2, PIIINP (ELISA) and HA (RIA) were measured from serum collected the day of the biopsy and every 3 months during the treatment. METAVIR score was used. Results: MMP-2 (r = 0.28; p < 0.01), TIMP- 1 (r = 0.42; p < 0.001), HA (r = 0.50; p < 0.001) and PIIINP (r = 0.62; p < 0.0001) were positively correlated with Metavir fibrosis, whereas MMP-1 (r = -0.34; r < 0.001) and MMP-2 (r = -0.24; r < 0.01) were negatively correlated. By multivariate analysis, PIIINP and MMP-1 were independently associated with fibrosis. A score calculated by combining these two variables (logistic regression function) was able to discriminate F0/F1 from F2/F3/F4 with a sensitivity of 60% and a specificity of 91% (ROC curves). During treatment, MMP-1 significantly increase and PIIINP as well as combined score significantly decrease in virological responders but not in non responders. Conclusion: Non invasive markers might be useful for estimating liver fibrosis. IFN and ribavirin reduce liver fibrosis in virological responders by decreasing fibrogenesis as well as increasing fibrolysis.
GRADING OF HEPATITIS C RELATED LIVER • TNON-INVASIVE • DISEASE: MICROBUBBLE-ENI'IANCED ULTRASOUND STUDIES Adrian Liml'2, Nayna Patel 1.2, Gavin Hamilton L2, Martin Blomiey 1, David Cosgrove 1, Graham Foster 2, Robert Goldin 3, Howard Thomas 2, Simon Taylor-Robinson 1.2.1Department of Imaging, Imperial College,
(4.9) respectively (Kruskal Wallis, p < 0.001). An AT < 20 s and CDT < 10 s was 100% sensitive for cirrhosis but only 69% and 72% specific respectively, as subjects with fibrosis sometimes showed early AT and CDT. 10 interferon-treated patients showed earlier arrival times than comparative untreated subjects. Conclusion: AT and CDT measurements hold promise in charaeterising HCV liver disease and are highly sensitive markers of cirrhosis. Treatment with interferon appears to prolong AT. This is important for disease monitoring and may be useful in assessing the efficacy of treatment regimes non-invasively and possibly replace repeat liver biopsy in some situations.
• TLOW 3RISK"OF]VERTICAL HEPATITIS C VIRUS (HCV) INFECTION IN A LARGE COHORT OF HUMAN IMMUNODEFICIENCY VIRUS (HIV)-SERONEGATIVE PREGNANT WOMEN Marina Losa l, Alicia Saez 2, Oreste Lo Iacono 3, Carlos Lozano 2, Carmelo Garcia-Monzon 1. l Hepatology Unit, UniversityHospital Santa
Cristina, Madrid; 2Serologyand Biochemistry Service, University Hospital Santa Cristina, Madrid, Spain; JGastmentemlogy Department, University of Palermo, Italy Given that conflicting results are currently available on vertical HCV transmission, the aims of this study were to 'determine the prevalence of chronic HCV infection in a large cohort of HIV-negative pregnant women and to analyze the rate of vertical HCV transmission, assessing the possible related risk factors. We prospectively recruited 20,612 consecutive unselected pregnant women who were referred to the Obstetric Unit of the University Hospital Santa Cristina. HCV-RNA was assessed in blood samples from anti-HCV+ mothers at first and third trimester of pregnancy and from their newborns at birth and after 6, 12, 24, and 36 months. Viral load and genotype were determined to all viremic women at the time of delivery as well as to viremic infants. Antibodies to HCV was detected in 119 mothers (0.57%), whereas HCV-RNA tested positive in 67% of anti-HCV+ women (n = 80). The median viral load of these viremic women, at time of delivery, was 335,300 copies per milliliter (range: 11,000-2,625,200), and the vast majority of them had a low viral load. HCV genotyping revealed subtype 1 in 60 cases (75%). Only 2 newborns tested positive for HCV-RNA immediately after birth (transmission rate: 2.4%), remaining positive during the entire follow-up. These vertically-infected babies had the same HCV genotype (lb) that their respective mothers. In conclusion, vertical HCV transmission is an infrequent event among HIV-negative HCV-infected women and, in the present study, no apparent risk factor influencing the transmission rate was identified.
~-~
GENOTYPE MIXTURES OF HEPATITIS B VIRUS IN PATIENTS TREATED WITH INTERFERON
Hammersmith Hospital, Du Cane Road, London; 2Departmentof Medicine, Imperial College, St. Mary's Hospital, Praed Street, London; 3Department of Histopathology, Imperial College, St. Mary's Hospital, Praed Street, London, UK
Charles Hannoun 1, Magnus Lindh 1, Kim Krogsgaard 2. 1Dept. of Clinical
Purpose: Non-invasive assessment of the severity of liver disease is difficult. We evaluated hepatic vein arrival times (AT) of an ultrasound microbubble agent and carotid-delay times (CDT) in grading liver disease in patients with chronic hepatitis C (HCV) infection. Materials/Method: 61 patients were studied. Grading of fibrosis (F) and inflammatory activity (NI) was carded out using the modified HA1 (Ishak) system. Patients were divided into mild hepatitis, (F < 3/6, NI < 4/18); moderate/severe hepatitis, (F > 2/6, NI > 3/18) and cirrhosis, (F = 6). Timeintensity curves of hepatic vein spectral Doppler signals and audio intensity from the carotid artery were analysed after an intravenous microbubble bolus. CDT was calculated as the difference between carotid and hepatic vein arrival times. Results: There was a monotonic decrease in the mean ATs (1 s.d.) and CDTs (1 s.d.) for mild, moderate/severe hepatitis and cirrhosis: 50.5 s (24.2), 33.6 s (26.1), 14.8 s (4.5) and 36.8 s (29), 18.7 s (22.6) and 5.8 s
kground: Little is known about co-infection with several hepatitis B virus genotypes, although previous reports of recombination and genotype shifts indicate that this should occur. Aim: To evaluate genotypic co-infection in HBV infection. Method: Using a genotypic core region PCR that identifies mixtures even if one genotype predominates, serum samples from before and after interferon treatment of 30 HBeAg positive patients were analysed. Results: Co-infection with genotype A and another genotype was found in 19 (63%) of 30 HBeAg positive patients treated with interferon. In 7 patients co-infection or genotype shifts were detectable also by direct sequencing or standard preS genotyping. In most cases, genotype changes were detected after a more than 2 log decrease or increase of the HBV DNA level. The presence of genotype mixtures did not significantly influence IFN response, but this has to be further analysed.
Virology, Goteborg University, Goteborg, Sweden; 2Clinical Research Unit, Hvidovre Hospital, Copenhagen, Denmark
Category 6: Viral hepatitis: clinical aspects
Condusion: Genotypic co-infection appears to be frequent. The finding may be relevant for tracing source of HBV infection and possibly for treatment.
[-~
MUTATIONS IN X REGION HAVE LITTLE IMPACT ON RESPONSE TO INTERFERON THERAPY FOR CHRONIC HEPATITIS B
Charles Hannoun i, Kim Krogsgaard 2, Peter Horal 1, Magnus Lindh 1.
1Department of Clinical Virology, Gtteborg University, Gtteborg, Sweden; 2Clinical Research Unit, Hvidovre Hospital, Copenhagen, Denmark
117
+ RBV 1-1.2 g/qd + AMA 200 mg/qd Group B: IFN alpha-2a 3 MU tiw sc + RBV 1-1.2 g/qd + AMA 200 mg/qd Group 3: IFN alpha-2a 3 MU tiw sc + RBV 1-1.2 g/qd R e s d t s : Of 268 so far enrolled pts, 55.5% were male, mean age was 51 yrs (range 20-65): genotype 1 was found in 57.6% of cases. Severe fibrosis or cirrhosis were found in 23.3%. The treatments were well tolerated; no major safety concerns were noted at the moment. One hundred and thirty pts have completed 3 mos therapy: at this time the percentages of pts with normal ALT levels were 51% in group A, 55% in group B, and 51% in group C. The rate of virological response was 54%, 49% and 42%, respectively. This study is ongoing and more data are needed to prove or disprove the value of triple therapy over the current standard therapy.
Background: Therapy for chronic hepatitis B with interferon-alpha (IFN) may result in viral clearance and hepatitis B e seroconversion in 30--40% of patients. It is still unclear if viral genetic variability influences response rates. However, certain core promoter mutations were recently associated with a better response to interferon. Aim" To analyse the importance of mutations in the core promoter for IFN response. Method: The entire X region, including the core promoter, of hepatitis B virus (HBV) from 26 HBeAg positive patients treated with IFN for 12 weeks, was sequenced. Serum samples from before treatment, at end-oftreatment and at follow-up of sustained and non-sustained responders were analysed. Most patients were of European origin and showed moderate aminotransferase elevation (mean 2.4 x upper limit of normal) and genotype A infection. Results: Before treatment, 21 patients had an X region identical with a consensus sequence of the corresponding genotype, and in the remaining 9 patients a mean of 1.3 mutations were found. After treatment, 1-4 new mutations (mean 1.8) had emerged in 5 patients. There was no association between specific mutations, or the number of mutations, and response to IFN. Conclusion: Analysis of X region/core promoter is of limited clinical value in HBeAg positive patients and emerging mutations in this region are not major determinants of IFN response.
I•-'•
A 3-ARM RANDOMIZED STUDY EVALUATING THE EFFICACY OFTRIPLE THERAPY OF PEGYLATED OR RECOMBINANT INTERFERON alpha-2a 3 MU BOTH COMBINED WITH RIBAVlRIN AND AMANTADINE VERSUS DUAL THERAPY OF RECOMBINANT INTERFERON alpha-2a PLUS RIBAVlRIN IN NAIVE PATIENTS WITH CHRONIC HCV HEPATITIS
Alessandra Mangia I , Giovanni Ricci2, Nicola Minerva 3, Marcello Persico 4, Vito Carretta 5, Donato Bacca 6, Franco Vinelli 7, Giuseppe Sogafi s, Vincenzo Guadagnino 9, Giovanni Maio, Angelo Andriulli 1. t Gastroenterology Division Hospital 'CSS' San
Giovanni Rotondo; 2Internal Medicine University 'La Sapienza' Roma; 3Medicine Division Hospital Canosa; 4Medicine Division Hospital 'Cotugno' Napoli; 5Medicine Division Hospital Venosa; 6Medicine Division Hospital Casarano; 7Gastroenterology Division Ospedali Riuniti Foggia," SMedicine Division Hospital 'SS.Annunziata' Taranto; 91nternal Medicine Division University Catanzaro Infectious Disease Division Hospital 'Rummo' Benevento, Italy
Background: We have previously shown that the addition of amantadine (AMA) to recombinant IFN alpha-2a was able to increase the efficacy of IFN monotherapy in pts with chronic hepatitis C (CHC). No data are available on naive pts to document whether triple therapy of AMA + ribavirin (RBV) and pegylated IFN (PEG 1FN) (group A) or recombinant IFN alpha-2a (IFN) (group B) improves the success rate over dual therapy with recombinant IFN + RIBA. Methods: 354 naive pts with serologically and biopy-proven CHC, persistently, elevated ALT and positive HCV RNA were randomized to a 3-arrn multicentre trial, as follows: Group A: PEG IFN alpha-2a 180 mcg sc qw
[-~
IL-10 ATA HAPLOTYPE AND 1ST INTERON IFN-GAMMA POLYMORPHISMS ARE MORE FREQUENT IN PTS WITH SPONTANEOUS HCV CLEARANCE
A. Mangia, R. Santoro, O. D'Apolito, A. Lacobellis, A. Andruilli.
1Gastroenterology IRCCS 'Casa Sollievo Della Sofferenza', San Giovanni Rotondo, Italy In chronic HCV infection, cytokines and their receptors play a critical role in regulating HCV immune response. The individual capacity for cytokine secretion has a major genetic component. Three different haplotype of IL-10 promoter region are associated with different serum levels of this cytokine, ATA haplotype correlating with low serum levels. Aim: to investigate the association between allelic polymorphisms of IFNy, IL-10, TNFc~, and spontaneous or IFN-induced (LTR) HCV RNA clearance. Methods: -1117, -854, -627 single nucleotide polymorphism (SNP) of IL-10 promoter by SSP and - 3 0 8 and - 2 3 8 SNP of TNFot by PCR and restriction enzymes digestion were evaluated. Microsatellite analysis of (ca) n at the 1st intron of IFNy was evaluated by PCR/acrylamide gel. Patient: 149 HCVRNA +ve pts: 105 with abnormal and 44 with normal ALT, and 78 HCV RNA - v e , 39 with past acute hepatitis and 39 long term responders to antivirals were evaluated. As controls 150 subjects from general population were assessed. Results are shown in the table HCV R N A + v e ALT> ALT N IL-10 ATA 3.3%* IFNyI25 bp 3.3%** TNFa -308AA 1% -238AA 1% *p = 0.07; ** p = 0.01
Spontaneous RNA clearance
LTR
Controls
6.6%
10.5%* 10.2%**
7.5% 3.8%
4.8% 4.8%
0 0
0 0
0 0
0 0
In conclusion: spontaneous HCV clearance is associated with ATA haplotype of IL-10 and polymorphisms of the 1st intron of IFNy.
~']
STUDY OF SEROLOGICAL MARKERS OF HBV IN POSTRANSFUSIONAL HEPATITIS C
Mafia Dolores Martin Arranz 1, Santos Del Campo Terron 2, Javier Moreno Garcia 2, Rafael Barcena Marugan 2. 1University Hospital La Paz.
Madrid; 2University Hospital Ramon y Cajal. Madrid, Spain Objetive: The aim of this study was to assess the influence of past virus B infection in the development of postransfusional chronic hepatitis C (HCV). Patients and Methods: We enrolled 184 patients with postransfusional HCV (previous transfusion without other causes of hepatitis), older than 18 years, liver biopsy at least 4 years after transfusion. None of them had HBsAg. All sera were collected at the time of liver biopsy. HBsAg, HBcAc, HBsAc were assessed by ELISA (Abbott). Serum and liver tissue