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HCV genotypes: Correlation with risk factors and response to interferon in Greek patients
410A
1213
AASLD ABSTRACTS
HCV GENOTYPES: CORRELATION WITH RISK FACTORS AND RESPONSE TO INTERFERON IN GREEK PATIENTS
1214
ROLE OF TRANSFERRIN IN THE TISSUE DELIVERY OF INJECTED S'Fe2+ AND S*Mn2+IN THE HYPOTRANSFERRINEMIC MOUSE. TK Dickinson, AG Devenvi & JR Conner. Penn State University College of Medicine, Milton S Hershey Medical Center, Hershey, PA 17033. Magnetic resonance signal hyperintensity of the globus pallidus has consistently been observed in cirrhotic patients. The etiology of this signal abnormality is unknown but increased pallidal manganese (Mn) concentrations have been reported in patients who died of hepatic coma. Basal ganglia Mn accumulation may be responsible for the extrapyramidal symptoms described in patients with hepatic encephalopathy. Mn is transported in the blood by transferrin (Tf). The role of Tf in the distribution of injected Mn and Fe was studied in the hypotransferrinemie (lip) mouse. Homozygous mutant animals (hh) have <1% of normal levels of circulating Tf and require supplemental injections of purified Tf. Heterozygote (Hh) Hp animals have approximately 50% of normal circulating Tf levels, require no supplemental Tf and live a normal life span. For this study, adult wild type (HH), Hh and hh mice were injected with 59Feor 54Mn. Animals received a single intraperitoneal injection and were sacrificed at 24 hours, 1 and 4 weeks. Analysis ineluded brain, liver, spleen, heart and plasma. Over the 4 week period, plasma, brain and heart levels of Fe and Mn remained constant whereas liver and spleen levels fell. Levels of brain Mn and Fe were equivalent in all animals at all times. In contrast, at 24 hrs., liver Mn levels were elevated in hh compared to HH and Hh animals (285+53 cpm/mg vs 212+14 and 90+24, respectively). At 1 and 4 weeks liver Mn levels were equivalent in all animals. Liver Fe levels were markedly elevated in mutant hh (817+-51 cpm/mg) animals compared to wild types (237+6) and heterozygotes (215+-22) at 24 hrs. These differences persisted at 1 wk. HH and Hh animals had similar Fe and Mn arguing that 50% of Tf levels are sufficient for normal Fe and Mn delivery to tissue. Alterations in both Fe and Mn delivery to liver and spleen in mutant animals is evidence for Tfs role in proper tissue targeting. Equivalent brain Mn and Fe level in HH, Hh, and hh mice suggest the existence of noa-Tf mediated delivery of Mn and Fe to the brain. Delineation of this mechanism will be important in understanding the mechanism of Mn accumulation in the basal ganglia in chronic liver disease.
1216
QUANTITATION OF INTRA-HEPATIC HCV-RNA ACCORDING TO H I S T O L O G Y AND T O GENOTYPES AFTER LIVER TRANSPLANTATION V DiMartino, C Frray, F Saurini, D Samuel, M Gigou, M Reyn~s, H Bismuth. Hepato-Biliary Surgery and Liver Transplant Unit, Pathology*, Paul Brousse Hospital.Villejuif. France
I. Diamantis 1, I. Vafiadis 2, E. Voskandou 2, J. Delladetsims 3 N.. J~ia(:li4. K. Gvr1. M. Batte£1av1. ~Policlinie of Internal Medicine, University Hospital Basel, Switzerland. 21st Dept. Propedeutic Medicine, ZPathology Dept. University of Athens Greece, 4Endotel AG Basel Switzerland. Back,qround: Genolypes of HCV have been identified, with a variable geographic distribution, different association to various risk factors and different response to IFN-alpha therapy. Aim__.__/:To investigate the prevalence of HCV genotypes among patients (pts) with chronic hepatitis C from different risk groups and to assess the response to IFN-alpha therapy. Patients and methods: HCV genotypes ware determined in 65 pts (18 pts after kidney transplantation, 16 with lhalassemia and 31 with unknown risk factors). Patients had elevated ALT (> 6months), histologically proven chronic hepatitis and ware anti-HCV positive. Genotypes ware determined by using the Inno-Lipa strip assay (Inogenetics). Results: Eleven out of 18 pts with kidney transplantation ware infected with genotype 3a, 4 with a mixture of genotypes, 2 with genotype lb, and 1 with genotype la. In 8 out of 16 thalassemic pts genotype 3a was found. Genotype lb was found in 3 pts, genotype 1a in 2 pts and genotype 2a, genotype 4a and infection with mixture of genotypes were found in the remaining 3 pts. Finally, 15 out of 31 pts with no risk factors showed infection with genotype 3a, 5/31 infection with a mixture of genotypes, 4/31 genotype lb, 3/31 genotype la, and 3/31 genotype 2a.ln all patients with mixed infection genotype 3a was found. 36 pts from the last twa groups received INF (3 Mio U/3x week) for one year. Half a year after the end of therapy biochemical, virological and histological responses to IFN-alpha therapy ware found in 11 out of 19 pts with genotype 3a (58%), in 3/5 with mixed infection, 2/4 with genotype lb, 2/5 with genotype 2a, 1/4 with genotype la, and 1/1 with genotype 4. Conclusions: 1. HCV genotype 3a is prevalent in Greek pts (68% of all pts). 2. There is no significant difference in genotypes among pts with different risk factors, 3. Although based on small numbers, pts with genotype 3a seem to respond better to therapy and mixed infection may not be associated with failure to IFN-alpha therapy.
1215
RETREATMENTAND INTERMITTENT MAINTENANCE THERAPY WITH INTERFERON FOR RELAPSING HEPATITIS C
Dickson Re, Drlscoll CJ. Ishitani MB. Oelsner D, Caldwell SH. Internal Medicine and Surgery, University of Virginia, Charlottesville, VA. Standard interferon u2b (Ifn) for hepatitis (HCV) has a 50% relapse rate. AIM: To assess the efficacy of a short course of Ifn followed by intermittent maintenance therapy in relapsing HCV. METHODS: 10 patients (pts) with HCV and prior response to Ifn with relapse were studied. The induction phas e consisted of a 2-4 month course of Ifn (3MU/3x/wk). The maintenance phase was begun if transaminases normalized with loss of HCV RNA at 2 months, or with a biochemical response with or without loss of HCV RNA at 4 months. The maintenance dose was 3 MU/3x/wk for one week per month until relapse or one year of therapy. R E S U L T S : 9 of 10 pts had a biochemical response - 8 within 2 months. 6 of 10 pts became PeR negative. 3 pts did not enter maintenance due to thrombocytopenia (1), worsening pulmonary status (1), and failure to reach biochemical or virologic response (1). 2 pts are currently in maintenance. Of the 5 pts completing the maintenance phase, 4 relapsed; at 1 month (2), two months (i), and three months (i). One pt remains in biochemical remission at 10 months, but had recurrence of viremia at 5 months. CONCLUSION: Reinduction of remission with a short course of Ifn and low dose intermittent maintenance therapy is feasible in relapsing HCV. However, the minimal effective dose to maintain remission has yet to be determined. C
HEPATOLOGY O c t o b e r 1995
A strong relation between genotypes lb and severity of recurrent liver disease due to HCV has been described after liver transplantation. However, the mechanism of such severity remains unknown. The aim of this study was to analyse in liver transplanted patients remaining serum HCV RNA positive, the relations between the level of intrahepatic replication of HCV, the severity of liver disease and the genotypes. Samples and methods: 70 post-transplant biopsies from 45 patients were available. Intrahepatic HCV RNA was quantitated by competitive PCR in the 5' non coding part of HCV RNA. Serial dilutions of cDNA were coamplified with the same amount of competitor and using a common set of primers. For all biopsie, a similar competitive method was used for the quantification of 28S ribosomal RNA permitting to take in account the rates of extraction and reverse transcription. Quantification was expressed in arbitrary unit (AU) of HCV competitor/AU of 28 S competitor. Results : Distribution of genotypes after LT was the following: type lb: 30; type la: 10 type 2a: 5. A significative relation was found between genotype 1 b and hepatitis on liver graft (22/30 vs 6/15 (p
1213
AASLD ABSTRACTS
HCV GENOTYPES: CORRELATION WITH RISK FACTORS AND RESPONSE TO INTERFERON IN GREEK PATIENTS
1214
ROLE OF TRANSFERRIN IN THE TISSUE DELIVERY OF INJECTED S'Fe2+ AND S*Mn2+IN THE HYPOTRANSFERRINEMIC MOUSE. TK Dickinson, AG Devenvi & JR Conner. Penn State University College of Medicine, Milton S Hershey Medical Center, Hershey, PA 17033. Magnetic resonance signal hyperintensity of the globus pallidus has consistently been observed in cirrhotic patients. The etiology of this signal abnormality is unknown but increased pallidal manganese (Mn) concentrations have been reported in patients who died of hepatic coma. Basal ganglia Mn accumulation may be responsible for the extrapyramidal symptoms described in patients with hepatic encephalopathy. Mn is transported in the blood by transferrin (Tf). The role of Tf in the distribution of injected Mn and Fe was studied in the hypotransferrinemie (lip) mouse. Homozygous mutant animals (hh) have <1% of normal levels of circulating Tf and require supplemental injections of purified Tf. Heterozygote (Hh) Hp animals have approximately 50% of normal circulating Tf levels, require no supplemental Tf and live a normal life span. For this study, adult wild type (HH), Hh and hh mice were injected with 59Feor 54Mn. Animals received a single intraperitoneal injection and were sacrificed at 24 hours, 1 and 4 weeks. Analysis ineluded brain, liver, spleen, heart and plasma. Over the 4 week period, plasma, brain and heart levels of Fe and Mn remained constant whereas liver and spleen levels fell. Levels of brain Mn and Fe were equivalent in all animals at all times. In contrast, at 24 hrs., liver Mn levels were elevated in hh compared to HH and Hh animals (285+53 cpm/mg vs 212+14 and 90+24, respectively). At 1 and 4 weeks liver Mn levels were equivalent in all animals. Liver Fe levels were markedly elevated in mutant hh (817+-51 cpm/mg) animals compared to wild types (237+6) and heterozygotes (215+-22) at 24 hrs. These differences persisted at 1 wk. HH and Hh animals had similar Fe and Mn arguing that 50% of Tf levels are sufficient for normal Fe and Mn delivery to tissue. Alterations in both Fe and Mn delivery to liver and spleen in mutant animals is evidence for Tfs role in proper tissue targeting. Equivalent brain Mn and Fe level in HH, Hh, and hh mice suggest the existence of noa-Tf mediated delivery of Mn and Fe to the brain. Delineation of this mechanism will be important in understanding the mechanism of Mn accumulation in the basal ganglia in chronic liver disease.
1216
QUANTITATION OF INTRA-HEPATIC HCV-RNA ACCORDING TO H I S T O L O G Y AND T O GENOTYPES AFTER LIVER TRANSPLANTATION V DiMartino, C Frray, F Saurini, D Samuel, M Gigou, M Reyn~s, H Bismuth. Hepato-Biliary Surgery and Liver Transplant Unit, Pathology*, Paul Brousse Hospital.Villejuif. France
I. Diamantis 1, I. Vafiadis 2, E. Voskandou 2, J. Delladetsims 3 N.. J~ia(:li4. K. Gvr1. M. Batte£1av1. ~Policlinie of Internal Medicine, University Hospital Basel, Switzerland. 21st Dept. Propedeutic Medicine, ZPathology Dept. University of Athens Greece, 4Endotel AG Basel Switzerland. Back,qround: Genolypes of HCV have been identified, with a variable geographic distribution, different association to various risk factors and different response to IFN-alpha therapy. Aim__.__/:To investigate the prevalence of HCV genotypes among patients (pts) with chronic hepatitis C from different risk groups and to assess the response to IFN-alpha therapy. Patients and methods: HCV genotypes ware determined in 65 pts (18 pts after kidney transplantation, 16 with lhalassemia and 31 with unknown risk factors). Patients had elevated ALT (> 6months), histologically proven chronic hepatitis and ware anti-HCV positive. Genotypes ware determined by using the Inno-Lipa strip assay (Inogenetics). Results: Eleven out of 18 pts with kidney transplantation ware infected with genotype 3a, 4 with a mixture of genotypes, 2 with genotype lb, and 1 with genotype la. In 8 out of 16 thalassemic pts genotype 3a was found. Genotype lb was found in 3 pts, genotype 1a in 2 pts and genotype 2a, genotype 4a and infection with mixture of genotypes were found in the remaining 3 pts. Finally, 15 out of 31 pts with no risk factors showed infection with genotype 3a, 5/31 infection with a mixture of genotypes, 4/31 genotype lb, 3/31 genotype la, and 3/31 genotype 2a.ln all patients with mixed infection genotype 3a was found. 36 pts from the last twa groups received INF (3 Mio U/3x week) for one year. Half a year after the end of therapy biochemical, virological and histological responses to IFN-alpha therapy ware found in 11 out of 19 pts with genotype 3a (58%), in 3/5 with mixed infection, 2/4 with genotype lb, 2/5 with genotype 2a, 1/4 with genotype la, and 1/1 with genotype 4. Conclusions: 1. HCV genotype 3a is prevalent in Greek pts (68% of all pts). 2. There is no significant difference in genotypes among pts with different risk factors, 3. Although based on small numbers, pts with genotype 3a seem to respond better to therapy and mixed infection may not be associated with failure to IFN-alpha therapy.
1215
RETREATMENTAND INTERMITTENT MAINTENANCE THERAPY WITH INTERFERON FOR RELAPSING HEPATITIS C
Dickson Re, Drlscoll CJ. Ishitani MB. Oelsner D, Caldwell SH. Internal Medicine and Surgery, University of Virginia, Charlottesville, VA. Standard interferon u2b (Ifn) for hepatitis (HCV) has a 50% relapse rate. AIM: To assess the efficacy of a short course of Ifn followed by intermittent maintenance therapy in relapsing HCV. METHODS: 10 patients (pts) with HCV and prior response to Ifn with relapse were studied. The induction phas e consisted of a 2-4 month course of Ifn (3MU/3x/wk). The maintenance phase was begun if transaminases normalized with loss of HCV RNA at 2 months, or with a biochemical response with or without loss of HCV RNA at 4 months. The maintenance dose was 3 MU/3x/wk for one week per month until relapse or one year of therapy. R E S U L T S : 9 of 10 pts had a biochemical response - 8 within 2 months. 6 of 10 pts became PeR negative. 3 pts did not enter maintenance due to thrombocytopenia (1), worsening pulmonary status (1), and failure to reach biochemical or virologic response (1). 2 pts are currently in maintenance. Of the 5 pts completing the maintenance phase, 4 relapsed; at 1 month (2), two months (i), and three months (i). One pt remains in biochemical remission at 10 months, but had recurrence of viremia at 5 months. CONCLUSION: Reinduction of remission with a short course of Ifn and low dose intermittent maintenance therapy is feasible in relapsing HCV. However, the minimal effective dose to maintain remission has yet to be determined. C
HEPATOLOGY O c t o b e r 1995
A strong relation between genotypes lb and severity of recurrent liver disease due to HCV has been described after liver transplantation. However, the mechanism of such severity remains unknown. The aim of this study was to analyse in liver transplanted patients remaining serum HCV RNA positive, the relations between the level of intrahepatic replication of HCV, the severity of liver disease and the genotypes. Samples and methods: 70 post-transplant biopsies from 45 patients were available. Intrahepatic HCV RNA was quantitated by competitive PCR in the 5' non coding part of HCV RNA. Serial dilutions of cDNA were coamplified with the same amount of competitor and using a common set of primers. For all biopsie, a similar competitive method was used for the quantification of 28S ribosomal RNA permitting to take in account the rates of extraction and reverse transcription. Quantification was expressed in arbitrary unit (AU) of HCV competitor/AU of 28 S competitor. Results : Distribution of genotypes after LT was the following: type lb: 30; type la: 10 type 2a: 5. A significative relation was found between genotype 1 b and hepatitis on liver graft (22/30 vs 6/15 (p