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Hepatitis B virus core gene sequence in patients with chronic hepatitis B from an HBV hyperendemic area
Poster Sessions
224
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HEPATITIS B VIRUS CORE GENE SEQUENCE IN PATIENTS WITH CHRONIC HEPATITIS B FROM AN HBV HYPERENDEMIC AREA
Loreta Kondili], Claudio Argentinil, Valentina La Sorsa 1, Paola Chionne ~, Angela Costantino I , Alfonso Mele 2, Mqurizia R. Brunetto 3, Maria Rapicetta I . llstituto Superiore Di Sanita ',
Laboratorio Di Virologia, Rome; 21stituto Superiore Di Sanita', Laboratorio Di Epidemiologia E Biostatistica, Rome; 30spedali Riuniti Di Santa Chiara, Pisa, Italy Background and Aims: In the Mediterranean region the m1986 mutation prevails in greater than 98% of HBeAg negative chronic hepatitis B patients (CHB). In this study the frequence of addittional mutations in the precore region was investigated in Albanian patients with CHB Methods: The hepatitis B virus precore/core gene from 40 patients with CHB was amplified by PCR and nucleotide sequences were analysed. Results and Conclussions: Sixteen HBeAg positive and 24 HBeAg negative sera were studied. All HBeAg negative sera showed m1896, and 16 (67%) had one or more addittional mutations (nucleotide: 1762, 1764, 1753, 1899 1907). Compared with wild viremia, patients with additional mutations were significantly older.. Eight of 16 I-IBeAg positive patients (50%) showed only wild-type and the remaining 8 (50%) showed heterogeneous viremia (in 2 patients wild and m 1896 and in 6, addittional mutations were observed). Although the mutations 1762T 1764A suppressed HBeAg no association was found between these and HBeAg negativity The additional mutations in both HBeAg positive and negative patients were always associated with higher HBV DNA levels, but was not found to be preferentially associated with active liver disease as evaluated by high levels of IgM anti core (>0.300), biochemical findings and/or histologic diagnosis
[ 8 ~ - ] IMMUNOMODULATION WITH INTERLEUKIN-12 IMPROVES THE EFFICACY OF THERAPEUTIC VACCINE IN PATIENTS WITH CHRONIC HEPATITIS B Hanns Loehr l, Benedikt Bubl z, Wulf Boecher 1, Sabine Herzog-Hauff I , Martin Hoffstedt 2, Peter Galle I . IL Department of Internal Medicine,
University Mainz~ Mainz; 2Genetis Institute, Martinsried, Germany Objective: The vaccination with HBV surface antigen induced cellular immune responses and viral clearance in some chronic HBV patients. IL-12 could induce virus specific T helper and eytotoxic T lymphocyte reactions (Th, TL) and have additional therapeutic efficacy. Methods: 20 chronic HBV carders (11 HBeAg+, 12 viral load > 1000 pgml) received six vaccinations (20 mcg I-IBsAg) within three months together with 300 ng/kg bw IL-12 sc. twice weekly. Liver cirrhosis and HCV coinfections were excluded. Clinical, serological and immunological data were monotored. Specific Th and CTL responses were analysed by ELISPOt and ELISA. Results: Moderate clinical therapy adverse effects included fatigue, fever, weight loss, myalgia and lymph node swelling. Eight patients showed negative HB-DNA and normal ALT levels after therapy, another patient showed >2 log decrease of viral loag and biochemical response. Two patients showed HBe seroconversion. Three months later two patients had relapsed. Male gender and high baseline viral load were associated with non-responsiveness. HBV core (HBcAg) specific Thl and peptide specific CTL were induced in all patients but their frequencies seemed to be higher in responders. Conclusions: Vaccine plus IL-12 therapy in chronic HBV infection was safe and effective even in unfavorable patients. Using IL-12 the peripheral tolerance could be broken i.e. in chronic HBV carriers with low to medium viral load.
~ 0 - ~ ULTRASONIC SCALER: POSSIBLE ROUTE FOR HEPATITIS C VIRUS TRANSMISSION Michelle Martinot-Peignoux 1, Patrick Marcellin2, Vanassa Esnault t, Jean Pierre Lezy 3, Dominique Valla2, Christian Vacher 3. ~INSERM U481,
Hopital Beaujon, Clichy; 2Service D'Hepatologie Hopital Beaujon, Clichy; 3Service Chirurgie Maxillo-Faciale et de Stomatologie, Hopital Beaujon, Clichy, France The route for hepatitis C virus infection is unknown for 30% of the infected subjects. The recommendations for heat treatment for ultra scalers are quite recent. Our aim was to evaluate the presence of HCV RNA in saliva and irrigation fluid of the ultra sonic scaler before and after decontamination in patients with chronic hepatitis C. Methods: We tested the presence of HCV RNA in the saliva before scaling and in the irrigation fluid before and after 10 minutes Polyhexamide decontamination in nine patients with chronic hepatitis C. A sensitive qualitative assay for the detection of HCV RNA was used (TMA, VERSANTTM HCV RNA qualitative assay, GEN-PROBE, BAYER). The specificity and sensitivity of the assay was assessed with serial dilution of the WHO standard. Results: The assay showed 100% of specificity and a sensitivity of 5 IU/ml. HCV RNA was present in 57%, 55%, and 11% of the saliva, irrigation fluids before and after decontamination, respectively. Median serum HCV RNA level was 380 x 103 IU/ml (range 83 to 3099 x 103 IU/ml). We found no correlation between the presence of HCV RNA in saliva, irrigation fluids before and after decontamination and serum HBV DNA levels. Conclusion: Our results confirm, a high prevalence (57%) of patients with chronic hepatitis C and detectable HCV RNA in saliva. HCV RNA is found in irrigation fluids before but even after decontamination, suggesting that scaling might have been a route for hepatitis C transmission before recent the use of heat treatment for ultra sealers.
[80-~ LONGITUDINAL ANALYSIS OF CHANGES IN EARLY HCV KINETICS FOLLOWING THE ADDITION OF RIBAVIRIN TO IFN MONOTHERAPY J.E.M. Medeiros-Filho I , A.U. Neumann 2, J.R.R. Pinho 3, I.M.V.G.C. Mello s, M.E Lemos 3, L.C. Da Silva 1, A.A. Laudann 1, F.J. Carrilho 1. 1Hepatology Branch, Department of Gastroenterology,
University of Sat Paulo School of Medicine; svirology Branch, Adolfo Lutz Institute, Sat Paulo, Brazil; 2Faculty of Life Sciences, Bar llan University, Ramat Gan, Israel Background: Ribavirin improves IFN sustained response rate, but cross sectional studies didnot show an effect on early viral kinetics. Methods: RBV (1 g/day) was added to IFN monotherapy (1 dose of 9 MU, 3 MU daily for 14 days followed by 3 MU each other day for 12 months) in 3 naive genotype 1 patients with positive PCR at third month. Also, 2 IFN monotherapy relapsers (genotype 1 and 2a) were retreated with IFN as above and RBV. HCV RNA was determined by COBAS Amplicor V2 daily during the first 14 days, then monthly. Results: An additional viral decline of 0.9 log IU/ml was observed following RBV addition in 2 of 3 patients. However, only one of them showed acceleration of decline slope from flat to 0.16/day and negative PCR 3 months after. Viral decline 24 hours after IFN + RBV re-treatment was somewhat higher (>1.9 and 2.1 log IU/ml) compared to previous IFN monotherapy in same patients (1.2 and 1.7, accordingly), although baseline viral level was the same as in first treatment. In first patient viremia became undetectable at 24 hours. The second patient had a rapid second phase slope (-0.26/day) comparable to that observed during the previous IFN monotherapy treatment. Conclusions: Longitudinal analysis of first phase viral decline indicates that Ribavirin may add a small anti-virai effect in blocking virion production, in agreement with previous results of Ribavirin monotherapy anti-viral effect (Pawlotsky et al, 2000). However, its effect on second phase decline during to IFN dally treatment remains unclear.
224
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HEPATITIS B VIRUS CORE GENE SEQUENCE IN PATIENTS WITH CHRONIC HEPATITIS B FROM AN HBV HYPERENDEMIC AREA
Loreta Kondili], Claudio Argentinil, Valentina La Sorsa 1, Paola Chionne ~, Angela Costantino I , Alfonso Mele 2, Mqurizia R. Brunetto 3, Maria Rapicetta I . llstituto Superiore Di Sanita ',
Laboratorio Di Virologia, Rome; 21stituto Superiore Di Sanita', Laboratorio Di Epidemiologia E Biostatistica, Rome; 30spedali Riuniti Di Santa Chiara, Pisa, Italy Background and Aims: In the Mediterranean region the m1986 mutation prevails in greater than 98% of HBeAg negative chronic hepatitis B patients (CHB). In this study the frequence of addittional mutations in the precore region was investigated in Albanian patients with CHB Methods: The hepatitis B virus precore/core gene from 40 patients with CHB was amplified by PCR and nucleotide sequences were analysed. Results and Conclussions: Sixteen HBeAg positive and 24 HBeAg negative sera were studied. All HBeAg negative sera showed m1896, and 16 (67%) had one or more addittional mutations (nucleotide: 1762, 1764, 1753, 1899 1907). Compared with wild viremia, patients with additional mutations were significantly older.. Eight of 16 I-IBeAg positive patients (50%) showed only wild-type and the remaining 8 (50%) showed heterogeneous viremia (in 2 patients wild and m 1896 and in 6, addittional mutations were observed). Although the mutations 1762T 1764A suppressed HBeAg no association was found between these and HBeAg negativity The additional mutations in both HBeAg positive and negative patients were always associated with higher HBV DNA levels, but was not found to be preferentially associated with active liver disease as evaluated by high levels of IgM anti core (>0.300), biochemical findings and/or histologic diagnosis
[ 8 ~ - ] IMMUNOMODULATION WITH INTERLEUKIN-12 IMPROVES THE EFFICACY OF THERAPEUTIC VACCINE IN PATIENTS WITH CHRONIC HEPATITIS B Hanns Loehr l, Benedikt Bubl z, Wulf Boecher 1, Sabine Herzog-Hauff I , Martin Hoffstedt 2, Peter Galle I . IL Department of Internal Medicine,
University Mainz~ Mainz; 2Genetis Institute, Martinsried, Germany Objective: The vaccination with HBV surface antigen induced cellular immune responses and viral clearance in some chronic HBV patients. IL-12 could induce virus specific T helper and eytotoxic T lymphocyte reactions (Th, TL) and have additional therapeutic efficacy. Methods: 20 chronic HBV carders (11 HBeAg+, 12 viral load > 1000 pgml) received six vaccinations (20 mcg I-IBsAg) within three months together with 300 ng/kg bw IL-12 sc. twice weekly. Liver cirrhosis and HCV coinfections were excluded. Clinical, serological and immunological data were monotored. Specific Th and CTL responses were analysed by ELISPOt and ELISA. Results: Moderate clinical therapy adverse effects included fatigue, fever, weight loss, myalgia and lymph node swelling. Eight patients showed negative HB-DNA and normal ALT levels after therapy, another patient showed >2 log decrease of viral loag and biochemical response. Two patients showed HBe seroconversion. Three months later two patients had relapsed. Male gender and high baseline viral load were associated with non-responsiveness. HBV core (HBcAg) specific Thl and peptide specific CTL were induced in all patients but their frequencies seemed to be higher in responders. Conclusions: Vaccine plus IL-12 therapy in chronic HBV infection was safe and effective even in unfavorable patients. Using IL-12 the peripheral tolerance could be broken i.e. in chronic HBV carriers with low to medium viral load.
~ 0 - ~ ULTRASONIC SCALER: POSSIBLE ROUTE FOR HEPATITIS C VIRUS TRANSMISSION Michelle Martinot-Peignoux 1, Patrick Marcellin2, Vanassa Esnault t, Jean Pierre Lezy 3, Dominique Valla2, Christian Vacher 3. ~INSERM U481,
Hopital Beaujon, Clichy; 2Service D'Hepatologie Hopital Beaujon, Clichy; 3Service Chirurgie Maxillo-Faciale et de Stomatologie, Hopital Beaujon, Clichy, France The route for hepatitis C virus infection is unknown for 30% of the infected subjects. The recommendations for heat treatment for ultra scalers are quite recent. Our aim was to evaluate the presence of HCV RNA in saliva and irrigation fluid of the ultra sonic scaler before and after decontamination in patients with chronic hepatitis C. Methods: We tested the presence of HCV RNA in the saliva before scaling and in the irrigation fluid before and after 10 minutes Polyhexamide decontamination in nine patients with chronic hepatitis C. A sensitive qualitative assay for the detection of HCV RNA was used (TMA, VERSANTTM HCV RNA qualitative assay, GEN-PROBE, BAYER). The specificity and sensitivity of the assay was assessed with serial dilution of the WHO standard. Results: The assay showed 100% of specificity and a sensitivity of 5 IU/ml. HCV RNA was present in 57%, 55%, and 11% of the saliva, irrigation fluids before and after decontamination, respectively. Median serum HCV RNA level was 380 x 103 IU/ml (range 83 to 3099 x 103 IU/ml). We found no correlation between the presence of HCV RNA in saliva, irrigation fluids before and after decontamination and serum HBV DNA levels. Conclusion: Our results confirm, a high prevalence (57%) of patients with chronic hepatitis C and detectable HCV RNA in saliva. HCV RNA is found in irrigation fluids before but even after decontamination, suggesting that scaling might have been a route for hepatitis C transmission before recent the use of heat treatment for ultra sealers.
[80-~ LONGITUDINAL ANALYSIS OF CHANGES IN EARLY HCV KINETICS FOLLOWING THE ADDITION OF RIBAVIRIN TO IFN MONOTHERAPY J.E.M. Medeiros-Filho I , A.U. Neumann 2, J.R.R. Pinho 3, I.M.V.G.C. Mello s, M.E Lemos 3, L.C. Da Silva 1, A.A. Laudann 1, F.J. Carrilho 1. 1Hepatology Branch, Department of Gastroenterology,
University of Sat Paulo School of Medicine; svirology Branch, Adolfo Lutz Institute, Sat Paulo, Brazil; 2Faculty of Life Sciences, Bar llan University, Ramat Gan, Israel Background: Ribavirin improves IFN sustained response rate, but cross sectional studies didnot show an effect on early viral kinetics. Methods: RBV (1 g/day) was added to IFN monotherapy (1 dose of 9 MU, 3 MU daily for 14 days followed by 3 MU each other day for 12 months) in 3 naive genotype 1 patients with positive PCR at third month. Also, 2 IFN monotherapy relapsers (genotype 1 and 2a) were retreated with IFN as above and RBV. HCV RNA was determined by COBAS Amplicor V2 daily during the first 14 days, then monthly. Results: An additional viral decline of 0.9 log IU/ml was observed following RBV addition in 2 of 3 patients. However, only one of them showed acceleration of decline slope from flat to 0.16/day and negative PCR 3 months after. Viral decline 24 hours after IFN + RBV re-treatment was somewhat higher (>1.9 and 2.1 log IU/ml) compared to previous IFN monotherapy in same patients (1.2 and 1.7, accordingly), although baseline viral level was the same as in first treatment. In first patient viremia became undetectable at 24 hours. The second patient had a rapid second phase slope (-0.26/day) comparable to that observed during the previous IFN monotherapy treatment. Conclusions: Longitudinal analysis of first phase viral decline indicates that Ribavirin may add a small anti-virai effect in blocking virion production, in agreement with previous results of Ribavirin monotherapy anti-viral effect (Pawlotsky et al, 2000). However, its effect on second phase decline during to IFN dally treatment remains unclear.