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Reduced sensitivity of HCV genotype 3 to αInterferon 2B (introna) when co-infected with genotype 1
GASTROENTEROLOGY Vol. 114, No. 4
A1314 AASLD ABSTRACTS
• L0471 REDUCED SENSITIVITY OF HCV GENOTYPE 3 TO c~INTERFERON 2B (INTRONA) WHEN CO-INFECTED WITH GENOTYPE 1. D.P. Nunes G.D. Offner, NH Afdhal. Section of Gastroenterology Boston University School of Medicine, Boston MA 02118. Introduction: Infection with more than one hepatitis C genotype has been reported with varying frequency. There are few data on the response rate or pattem of response in this subgroup of patients. The aim of the present study was to assess the pattern of genotype response both during and after treatment in a cohort of patients with mixed genotype (MG) infection. Methods: Data from patients who had previously participated in a 24 week escalating dose interferon (INTRON A) study were analysed. Genotyping in patients with MG infection was performed at baseline, months 2, 4, and 6 of treatment and at 4 months post therapy~ Genotyping was performed with biotinylated PCR products using a novel covalent reverse dot blot assay. Oligonucleotide probes were modified to include a 5' amino group which was covalently bound to a membrane support (Biodyne C). Because of the uncharged nature of this membrane no prehybridisation step or inclusion of foreign DNA during hybridisation is required. Results: 12 of 61 patients were found to have a MG infection, 2 with genotypes la and 3, and 10 with types lb and 3. Patients with MG infection were similar in age and sex distribution (mean age 41.2 yrs, M:F 8:4) to those with SG infection (mean age 40.3 M:F 39:10). There was a trend to a higher prevalence of intravenous drug abusers in the MG population 75% vs 53% but this did not reach statistical significance (p=0.2). Baseline biochemical and histological data were also similar; mean ALT 116 vs 113 iu/L, cirrhosis 2/12 vs 3/47 in the mixed and single genotype groups respectively. Response defined by either a transient or permanent loss in HCV RNA was observed in 6/12 (2 sustained) MG infected patients and in 24/47 (7/47 sustained) SG infected patients. Only one of the 12 patients lost one (type 3) but not the other genotype (lb) during therapy. He later relapsed on discontinuation of therapy. By comparison in the SG infected group of the 6 patients infected with genotype 3 alone, 4 had a sustained viral response (p = 0.02). Conelusions: The reverse dot blot assay is a convenient and robust method for HCV genotyping. Despite the small numbers, mixed geonotype infection appears to be more common in patients with multiple parenteral exposures but is not associated with more severe liver disease. Patients with mixed infections are unlikely to clear type 3 infection when dually infected with genotype 1, despite the favourable response profile for genotype 3. Supported by Schering Plough Incorporated. L0472 ISOLATION AND CHARACTERIZATION OF A NEW HEPATIC EPITHELIAL-LIKE CELL LINE (AKN-1) FROM A NORMAL LIVER. Nussler AK, Vergani G, Gollin SM, Gansauge S, Morris SM, Jr., Demetris AJ, Beger HG, Strom SC. Departments of Pathology, Molecular Genetics and Biochemistry, Human Genetics and the Pittsburgh Cancer Institute, University of Pittsburgh, Pittsburgh, USA, General Surgery, and Anatomy, University of Ulm, Ulm, Germany. Introduction/Aim: Significant knowledge has been gained on various aspects of human liver diseases. Besides clinical studies, basic research has examined hepatocellular functions, such as drug hepatoxicity, regeneration, and malignant transformation. However, isolation and characterization of human liver cell lines are difficult due to limited material and poor growth in cell culture. In this study, we present the isolation, culture conditions and characterization of a new epithelial-like liver cell line (AKN-1) with characteristics of normal liver and hepatocellular carcinoma. Methods: Cells were obtained after enzymatic digestion, followed by a Percoll gradient with various densities. Cells at the interface of 30 and 20% were placed into culture in the presence of 25 ng/ml EGF and cell growth observed within ten days. Results; AKN-1 cells stain positively with antibodies to epithelial cytokeratin polypetides CK 8, 18, and 19. In addition, the cells express the anti-human epithelial related antigen (MOC-31), the hemopoetic growth factor, stern cell factor, and its receptor, c-kit, but stained negative for albumin and factor 8. RNA derived from the cells scored positively for argininosuccinatesynthetase and tyrosine-aminotransferase. Cytoge-netic characterization revealed rare breakpoints in chromosome 2, which have not been reported in liver cells. Since these cells showed also tumor growth in nude mice, we investigated the presence of adhesion molecules. FACS-scan-analysis showed expression of ICAM-J/CD54 and CD44std./CD44v6, which may be important in tumor cell invasion and metastasis. Conclusiont Therefore, we conclude that this new cell line may permit the control and examination of various liver specific functions. • L0473 ASSESSMENT OF FATIGUE IN CHRONIC LIVER DISEASE. J. SI Obhrai and B. S. Anand. Baylor College of Medicine, and V. A. Medical Center, Houston, Texas. Background: It is a common clinical impression that fatigue is a frequent and
often times a predominant symptom in chronic hepatitis C virus (HCV) infection, and is believed to have a significant impact on the quality of life in these patients. However, despite its obvious importance very little work has been done to elucidate the relationship between fatigue, psychological
disturbances and liver disease. Methods: Three groups of subjects were included in the study: a) patients with chronic HCV infection, b) chronic systemic diseases not related to liver (non-liver controls), and c) healthy volunteers. All subjects were administered investigator-assisted questionnaires designed to assess the presence and severity of fatigue and psychological abnormalities. The person performing the interview was unaware of the clinical diagnosis of the patients. Results: A total of 117 subjects consisting of 46 patients with chronic HCV infection, 36 with nonliver diseases and 35 healthy volunteers were included in the study. Patients with HCV infection and non-liver controls had significantly greater mean ( _+ SD) fatigue scores (133 ± 27 and 130 ± 37, respectively) compared to healthy subjects (101 ± 32; p=0.001 for both), but there was no difference between patients with HCV infection and non-liver chronic diseases. However, relief from fatigue (with rest) was less effective in HCV infection compared to non-liver controls (p=0.03). Both the patient groups had higher scores for psychological disturbances compared to healthy subjects. Depression (20 ± 13 vs 7 ± 7; p<0.001), and feelings of anger and hostility (17 ± 10 vs 7 ± 7; p<0.002) were more prominent in HCV patients compared to non-liver patients. Conclusions: These results indicate that fatigue and psychological disturbances occur frequently in chronic systemic disorders, irrespective of the cause. Contrary to the general impression, HCV infection is not associated with greater fatigue compared to other chronic diseases. However, the fatigue seen in HCV infection is more intransigent, responding poorly to relieving factors such as rest. Moreover, HCV infected patients are more depressed and have greater feelings of anger and hostility compared to those with other chronic diseases. Our findings are important because proper management of these symptoms may have a favorable impact on the quality of life of HCV infected patients. L0474
EARLY DETECTION OF HEMOBILIA FOLLOWING PERCUTANEOUS ETHANOL INJECTION THERAPY FOR HEPATOCELLULAR CARCINOMA: "HEMOBILIA SIGN" OF GALLBLADDER. S Obi, S Shiina, M Imamura, T Teratani, K Hamamura, S Sato, H Yoshida, Y Koike, Y Dan, M Akamatsu, T Goto, G Okudaira, N Kato, Y Niwa, Y Shiratori and M Omata. Department of Medicine 0I), University of Tokyo, Tokyo, Japan. OBJECTIVE Hemobilia is often an iatrogenic injury caused by such procedures as ethanol injection. We report the results of a study on the detection of ethanol injection-induced hemobilia in 365 patients with hepatocellular carcinoma undergoing ethanol injection therapy. SUBJECTS AND METHODS We began to analyze the occurrence of hemobilia in January 1995 to find a better way of preventing it. Analysis of clinical, biochemical and imaging data revealed that changes in the gallbladder as imaged by ultrasonography, namely, the rapid appearance of echogenic material in the gallbladder lumen, the "hemobilia sign," was a very useful indicator for early detection of hemobilia. In January 1996 we began a prospective study on the early detection of hemobilia based on the "bemobilia sign." RESULTS The incidence of hemobilia in 1996, 8 of 968 sessions (0.7 %), was not different from that in 1995, 7 of 1066 (0.8 %). However the mean period of time from ethanol injection to the diagnosis was only 0.3 ± 0.4 days in the1996 group as compared to 2.8 ± 2.1 days in the 1995 group (p<0.001). The mean duration of jaundice in the 1996 group (4.3 days) was significantly shorter than in the 1995 group (40.0 days) (p<0.05). CONCLUSION Early diagnosis of hemobilia by simply monitoring the gallbladder lumen by ultrasonography to detect the "hemobilia sign" may minimize morbidity due to hemobilia related interventional treatment. L0475
ULTRASOUND DETECTED PERIHEPATIC LYMPHADENOPATHY AND RESPONSE TO INTERFERON ALPHA IN CHRONIC HEPATITIS C. J. Ocken~a. MD: H. Wedemeyer, MD; H. Frank, HL. Tillmann, MD; A. Schiller, MD; M Caselitz, MD; JS Bleck, MD; M Gebel, MD; K Btiker, MD; C Trautwein, MD; MP Manns; MD. Department of Gastroenterologyand Hepatology, Medizinische Hochschule Hannover; Germany. Recently it was shown that perihepatic lymphadenopathy (PHL) correlates with the histological activity in chronic hepatitis C. However, it is unclear whether there is a correlation between the response to interferon alpha and the PHL. METHODS: We examined 69 patients who had been treated with interferon alpha for hepatitis C. Prior to treatment all patients had been examined by high resolution ultrasonography (5 Mhz). Twenty-five patients had follow up ultrasound scans during the course of the treatment, and 16 of these after completion of treatment. We introduced a grading-system of the PHL based on size and number of lymph nodes (grade I, II, III). RESULTS: Classification of PHL prior to treatment revealed 22 patients with PHL I, 25 with grade II and 22 with grade IlL There were no significant correlation between PHL and HCV-genotype, viremia, liver function tests or histological grading and staging. In patients with PHL grade I primary response was more often observed compared to PHL grade II and III: 55%, 40%, and 41%, respectively. Increase of PHL during treatment was significantly associated with non-response and was detected in 7/14 non-responders compared to 1/11 primary responders (p<0,05).
A1314 AASLD ABSTRACTS
• L0471 REDUCED SENSITIVITY OF HCV GENOTYPE 3 TO c~INTERFERON 2B (INTRONA) WHEN CO-INFECTED WITH GENOTYPE 1. D.P. Nunes G.D. Offner, NH Afdhal. Section of Gastroenterology Boston University School of Medicine, Boston MA 02118. Introduction: Infection with more than one hepatitis C genotype has been reported with varying frequency. There are few data on the response rate or pattem of response in this subgroup of patients. The aim of the present study was to assess the pattern of genotype response both during and after treatment in a cohort of patients with mixed genotype (MG) infection. Methods: Data from patients who had previously participated in a 24 week escalating dose interferon (INTRON A) study were analysed. Genotyping in patients with MG infection was performed at baseline, months 2, 4, and 6 of treatment and at 4 months post therapy~ Genotyping was performed with biotinylated PCR products using a novel covalent reverse dot blot assay. Oligonucleotide probes were modified to include a 5' amino group which was covalently bound to a membrane support (Biodyne C). Because of the uncharged nature of this membrane no prehybridisation step or inclusion of foreign DNA during hybridisation is required. Results: 12 of 61 patients were found to have a MG infection, 2 with genotypes la and 3, and 10 with types lb and 3. Patients with MG infection were similar in age and sex distribution (mean age 41.2 yrs, M:F 8:4) to those with SG infection (mean age 40.3 M:F 39:10). There was a trend to a higher prevalence of intravenous drug abusers in the MG population 75% vs 53% but this did not reach statistical significance (p=0.2). Baseline biochemical and histological data were also similar; mean ALT 116 vs 113 iu/L, cirrhosis 2/12 vs 3/47 in the mixed and single genotype groups respectively. Response defined by either a transient or permanent loss in HCV RNA was observed in 6/12 (2 sustained) MG infected patients and in 24/47 (7/47 sustained) SG infected patients. Only one of the 12 patients lost one (type 3) but not the other genotype (lb) during therapy. He later relapsed on discontinuation of therapy. By comparison in the SG infected group of the 6 patients infected with genotype 3 alone, 4 had a sustained viral response (p = 0.02). Conelusions: The reverse dot blot assay is a convenient and robust method for HCV genotyping. Despite the small numbers, mixed geonotype infection appears to be more common in patients with multiple parenteral exposures but is not associated with more severe liver disease. Patients with mixed infections are unlikely to clear type 3 infection when dually infected with genotype 1, despite the favourable response profile for genotype 3. Supported by Schering Plough Incorporated. L0472 ISOLATION AND CHARACTERIZATION OF A NEW HEPATIC EPITHELIAL-LIKE CELL LINE (AKN-1) FROM A NORMAL LIVER. Nussler AK, Vergani G, Gollin SM, Gansauge S, Morris SM, Jr., Demetris AJ, Beger HG, Strom SC. Departments of Pathology, Molecular Genetics and Biochemistry, Human Genetics and the Pittsburgh Cancer Institute, University of Pittsburgh, Pittsburgh, USA, General Surgery, and Anatomy, University of Ulm, Ulm, Germany. Introduction/Aim: Significant knowledge has been gained on various aspects of human liver diseases. Besides clinical studies, basic research has examined hepatocellular functions, such as drug hepatoxicity, regeneration, and malignant transformation. However, isolation and characterization of human liver cell lines are difficult due to limited material and poor growth in cell culture. In this study, we present the isolation, culture conditions and characterization of a new epithelial-like liver cell line (AKN-1) with characteristics of normal liver and hepatocellular carcinoma. Methods: Cells were obtained after enzymatic digestion, followed by a Percoll gradient with various densities. Cells at the interface of 30 and 20% were placed into culture in the presence of 25 ng/ml EGF and cell growth observed within ten days. Results; AKN-1 cells stain positively with antibodies to epithelial cytokeratin polypetides CK 8, 18, and 19. In addition, the cells express the anti-human epithelial related antigen (MOC-31), the hemopoetic growth factor, stern cell factor, and its receptor, c-kit, but stained negative for albumin and factor 8. RNA derived from the cells scored positively for argininosuccinatesynthetase and tyrosine-aminotransferase. Cytoge-netic characterization revealed rare breakpoints in chromosome 2, which have not been reported in liver cells. Since these cells showed also tumor growth in nude mice, we investigated the presence of adhesion molecules. FACS-scan-analysis showed expression of ICAM-J/CD54 and CD44std./CD44v6, which may be important in tumor cell invasion and metastasis. Conclusiont Therefore, we conclude that this new cell line may permit the control and examination of various liver specific functions. • L0473 ASSESSMENT OF FATIGUE IN CHRONIC LIVER DISEASE. J. SI Obhrai and B. S. Anand. Baylor College of Medicine, and V. A. Medical Center, Houston, Texas. Background: It is a common clinical impression that fatigue is a frequent and
often times a predominant symptom in chronic hepatitis C virus (HCV) infection, and is believed to have a significant impact on the quality of life in these patients. However, despite its obvious importance very little work has been done to elucidate the relationship between fatigue, psychological
disturbances and liver disease. Methods: Three groups of subjects were included in the study: a) patients with chronic HCV infection, b) chronic systemic diseases not related to liver (non-liver controls), and c) healthy volunteers. All subjects were administered investigator-assisted questionnaires designed to assess the presence and severity of fatigue and psychological abnormalities. The person performing the interview was unaware of the clinical diagnosis of the patients. Results: A total of 117 subjects consisting of 46 patients with chronic HCV infection, 36 with nonliver diseases and 35 healthy volunteers were included in the study. Patients with HCV infection and non-liver controls had significantly greater mean ( _+ SD) fatigue scores (133 ± 27 and 130 ± 37, respectively) compared to healthy subjects (101 ± 32; p=0.001 for both), but there was no difference between patients with HCV infection and non-liver chronic diseases. However, relief from fatigue (with rest) was less effective in HCV infection compared to non-liver controls (p=0.03). Both the patient groups had higher scores for psychological disturbances compared to healthy subjects. Depression (20 ± 13 vs 7 ± 7; p<0.001), and feelings of anger and hostility (17 ± 10 vs 7 ± 7; p<0.002) were more prominent in HCV patients compared to non-liver patients. Conclusions: These results indicate that fatigue and psychological disturbances occur frequently in chronic systemic disorders, irrespective of the cause. Contrary to the general impression, HCV infection is not associated with greater fatigue compared to other chronic diseases. However, the fatigue seen in HCV infection is more intransigent, responding poorly to relieving factors such as rest. Moreover, HCV infected patients are more depressed and have greater feelings of anger and hostility compared to those with other chronic diseases. Our findings are important because proper management of these symptoms may have a favorable impact on the quality of life of HCV infected patients. L0474
EARLY DETECTION OF HEMOBILIA FOLLOWING PERCUTANEOUS ETHANOL INJECTION THERAPY FOR HEPATOCELLULAR CARCINOMA: "HEMOBILIA SIGN" OF GALLBLADDER. S Obi, S Shiina, M Imamura, T Teratani, K Hamamura, S Sato, H Yoshida, Y Koike, Y Dan, M Akamatsu, T Goto, G Okudaira, N Kato, Y Niwa, Y Shiratori and M Omata. Department of Medicine 0I), University of Tokyo, Tokyo, Japan. OBJECTIVE Hemobilia is often an iatrogenic injury caused by such procedures as ethanol injection. We report the results of a study on the detection of ethanol injection-induced hemobilia in 365 patients with hepatocellular carcinoma undergoing ethanol injection therapy. SUBJECTS AND METHODS We began to analyze the occurrence of hemobilia in January 1995 to find a better way of preventing it. Analysis of clinical, biochemical and imaging data revealed that changes in the gallbladder as imaged by ultrasonography, namely, the rapid appearance of echogenic material in the gallbladder lumen, the "hemobilia sign," was a very useful indicator for early detection of hemobilia. In January 1996 we began a prospective study on the early detection of hemobilia based on the "bemobilia sign." RESULTS The incidence of hemobilia in 1996, 8 of 968 sessions (0.7 %), was not different from that in 1995, 7 of 1066 (0.8 %). However the mean period of time from ethanol injection to the diagnosis was only 0.3 ± 0.4 days in the1996 group as compared to 2.8 ± 2.1 days in the 1995 group (p<0.001). The mean duration of jaundice in the 1996 group (4.3 days) was significantly shorter than in the 1995 group (40.0 days) (p<0.05). CONCLUSION Early diagnosis of hemobilia by simply monitoring the gallbladder lumen by ultrasonography to detect the "hemobilia sign" may minimize morbidity due to hemobilia related interventional treatment. L0475
ULTRASOUND DETECTED PERIHEPATIC LYMPHADENOPATHY AND RESPONSE TO INTERFERON ALPHA IN CHRONIC HEPATITIS C. J. Ocken~a. MD: H. Wedemeyer, MD; H. Frank, HL. Tillmann, MD; A. Schiller, MD; M Caselitz, MD; JS Bleck, MD; M Gebel, MD; K Btiker, MD; C Trautwein, MD; MP Manns; MD. Department of Gastroenterologyand Hepatology, Medizinische Hochschule Hannover; Germany. Recently it was shown that perihepatic lymphadenopathy (PHL) correlates with the histological activity in chronic hepatitis C. However, it is unclear whether there is a correlation between the response to interferon alpha and the PHL. METHODS: We examined 69 patients who had been treated with interferon alpha for hepatitis C. Prior to treatment all patients had been examined by high resolution ultrasonography (5 Mhz). Twenty-five patients had follow up ultrasound scans during the course of the treatment, and 16 of these after completion of treatment. We introduced a grading-system of the PHL based on size and number of lymph nodes (grade I, II, III). RESULTS: Classification of PHL prior to treatment revealed 22 patients with PHL I, 25 with grade II and 22 with grade IlL There were no significant correlation between PHL and HCV-genotype, viremia, liver function tests or histological grading and staging. In patients with PHL grade I primary response was more often observed compared to PHL grade II and III: 55%, 40%, and 41%, respectively. Increase of PHL during treatment was significantly associated with non-response and was detected in 7/14 non-responders compared to 1/11 primary responders (p<0,05).