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Mutations of the core promoter and response to interferon treatment in chronic replicative hepatitis B
General session 5
MUTATIONS OF THE CORE PROMmER AND RESPONSE TO INTERFERON TREATMENT IN CHRONIC REPLICATIVE HEPATITIS B A. Erhardt’. II. Reineke’, D. Blondin’. W.H. Gerlicha, 0. Adams3. T. Heintges’. C. Niederau, D. Haussinseri ‘Division of Gastroenterology, Hepatology and Infectious Diseases, Heinrich-Heine-University, Dttssledorf, Germany. ‘Institute of Medical Virology, Justus-Leibig-University, Giessen, Germany. ‘Institute of Medical Virology, Heinrich-Heine-University, Dttsseldorf, Germany. Background: In chronic replicative hepatitis B the signiIlcance of mutations in the basic core promoter (BCP), core upstream regulatory sequences (CURS) and negative regulatory element (NRE) for response to interferon (IFN) is unknown Methods: A sequence analysis of the NRE. CURS, BCP and precore region was performed from sera of 96 patients with chronic replicative hepatitis B (64 HBeAg-positive, 32 HBeAgnegative) treated with alpha-IFN. Results: The overall sustained response (SR) rate to IFN was 30% with no significant dierence between HBeAgpositive and HBeAg-negative patients. IFN responsiveness correlated to HBV DNA levels, HBsAg levels, the number of mutations in the complete BCP, especially nt region 1753 to 1766 and mutations at nt 1762 and 1764. In HBeAg-positive hepatitis SR to IFN was associated with a high number of mutations in the BCP (pQ.04) and nucleotide region 17531766 (pCO.015) as well as mutations at nucleotide 1764 (p
36
IS T(14;18)(q32;q21)TRANSLOCATION A HALLMARK OF HCV-RE LATED MIXED CRYOGLOBULINEMI.4? E Giannelli. C. Ferri’. M.E. Marrocchi. L. La Civita’. G. Loneombardo’. C. Giannini. A. Puliti. A.L. Giallombardo. M. Monti. P Gentilini. A.L. Zieneao Department of Internal Medicine, University of Florence, Florence, Italy. *Department of Internal Medicine, University of Pisa, Pisa, Italy. A pathogenetic roleof HCV infection has been definitively cstablishcd for mixed cryoglobulinemia (MC). HCV may b-e responsiblefor cloncl B-cell expansion underlying tbe MC. Preliminary data we obtained suggest c role of t(14:18) in the pathopnesis of HCV-r&ted B-cell lymphoprolifcmtivc disordm (LPDs). T(14;lS) tmnslocation (bcl-2 recombination) is interpreted as an error during V(D)1 gene rcanangement proin Ig gents and Icads to anti-cpoptotic bcl-2 oncoprotcin overexpression in B-cell. The aim of this study was to evahmtc the oworrcncc of both T(14;lS) and bcl-2 expression in peripheral blood mononuclear cells(PBMC) from HCV-positive MC patients. Patients and Methcds. Four groups of patients wcrc consecutively cvsluated: a) 25 HCV-rclatcd MC (M/F: 10/15. mean age 64+9SD yrr); 13 with type 11and I2 type III MC; 3 cases cvolti into a fmnk B-cell lymphoma; b) 25 pts with cbmnic HCV infection without MC, MiF IS/lo, meao age 51+11SD yrs; c) 25 pts with chronic HEN cod/or HDV infection, M/F 16/9, mean age 56 +1 ISD ITS; and d) 25 pts with HCV-negative chronic rheumatic disordas or chronic infection by non hepctotxopic agents, MiF 14/l 1, mean age 54.2+9SD yrs. Both t(14;18) and bcl-2 expression were evabmted in PBMC by “ncskd” bcl-2/JH PCR with diict sequencing of amplification products and by Westan blot analysis and immonoprccipitation. Results. T(14;lS) wcs deteoted in 18 (72%)).37 (32%), 1 (4%) cod 2 (8%) pts from groups a, b, c, and d rcsptxtivcly (fl.01). All PBMC samples with t(14;lS) ovcrcxpressed b&2, these findings did not corrclctc with the prcscnc-ckbscnce of monoclonal Ifl component in the clyoprccipitatcs (MC type II/III), as well as with various cliiica-epidcmiological pwamctcrs; however, a trend toward a higher frequency of bcl-2 recanbination and ovcrcxpressioa was obscrvcd in tvw II MC (83%) es well cs in older ots and more.scvcre liver dkasc. In addition t($lS) wcs d&t& in 2/3 MC pts &b complicating malignant B-cell lymphoma Discussion. The significantly high frequency of t(14;18) in HCV-n&ted MC pts, suggests that HCV may play c role in the multistep benign end mcligoant lymphopmlifemtion. Tbe inhibition of B-cell apoptosis by bcl-2 oncogcnc should rcprcsent the early step of such pathogenctic pmccss. The detection of t(14;lS) also in polyclonal, type III MC and in c rclcvcnt proportion of patients with chronic HCV infection without MC further supports this hypothesis. Finally, the detection of t(14; 18) in HCV infected patients may be a predictive factor for the development of MC and/or other lymphoprolifcmtive disorders.
MUTATIONS OF THE CORE PROMmER AND RESPONSE TO INTERFERON TREATMENT IN CHRONIC REPLICATIVE HEPATITIS B A. Erhardt’. II. Reineke’, D. Blondin’. W.H. Gerlicha, 0. Adams3. T. Heintges’. C. Niederau, D. Haussinseri ‘Division of Gastroenterology, Hepatology and Infectious Diseases, Heinrich-Heine-University, Dttssledorf, Germany. ‘Institute of Medical Virology, Justus-Leibig-University, Giessen, Germany. ‘Institute of Medical Virology, Heinrich-Heine-University, Dttsseldorf, Germany. Background: In chronic replicative hepatitis B the signiIlcance of mutations in the basic core promoter (BCP), core upstream regulatory sequences (CURS) and negative regulatory element (NRE) for response to interferon (IFN) is unknown Methods: A sequence analysis of the NRE. CURS, BCP and precore region was performed from sera of 96 patients with chronic replicative hepatitis B (64 HBeAg-positive, 32 HBeAgnegative) treated with alpha-IFN. Results: The overall sustained response (SR) rate to IFN was 30% with no significant dierence between HBeAgpositive and HBeAg-negative patients. IFN responsiveness correlated to HBV DNA levels, HBsAg levels, the number of mutations in the complete BCP, especially nt region 1753 to 1766 and mutations at nt 1762 and 1764. In HBeAg-positive hepatitis SR to IFN was associated with a high number of mutations in the BCP (pQ.04) and nucleotide region 17531766 (pCO.015) as well as mutations at nucleotide 1764 (p
36
IS T(14;18)(q32;q21)TRANSLOCATION A HALLMARK OF HCV-RE LATED MIXED CRYOGLOBULINEMI.4? E Giannelli. C. Ferri’. M.E. Marrocchi. L. La Civita’. G. Loneombardo’. C. Giannini. A. Puliti. A.L. Giallombardo. M. Monti. P Gentilini. A.L. Zieneao Department of Internal Medicine, University of Florence, Florence, Italy. *Department of Internal Medicine, University of Pisa, Pisa, Italy. A pathogenetic roleof HCV infection has been definitively cstablishcd for mixed cryoglobulinemia (MC). HCV may b-e responsiblefor cloncl B-cell expansion underlying tbe MC. Preliminary data we obtained suggest c role of t(14:18) in the pathopnesis of HCV-r&ted B-cell lymphoprolifcmtivc disordm (LPDs). T(14;lS) tmnslocation (bcl-2 recombination) is interpreted as an error during V(D)1 gene rcanangement proin Ig gents and Icads to anti-cpoptotic bcl-2 oncoprotcin overexpression in B-cell. The aim of this study was to evahmtc the oworrcncc of both T(14;lS) and bcl-2 expression in peripheral blood mononuclear cells(PBMC) from HCV-positive MC patients. Patients and Methcds. Four groups of patients wcrc consecutively cvsluated: a) 25 HCV-rclatcd MC (M/F: 10/15. mean age 64+9SD yrr); 13 with type 11and I2 type III MC; 3 cases cvolti into a fmnk B-cell lymphoma; b) 25 pts with cbmnic HCV infection without MC, MiF IS/lo, meao age 51+11SD yrs; c) 25 pts with chronic HEN cod/or HDV infection, M/F 16/9, mean age 56 +1 ISD ITS; and d) 25 pts with HCV-negative chronic rheumatic disordas or chronic infection by non hepctotxopic agents, MiF 14/l 1, mean age 54.2+9SD yrs. Both t(14;18) and bcl-2 expression were evabmted in PBMC by “ncskd” bcl-2/JH PCR with diict sequencing of amplification products and by Westan blot analysis and immonoprccipitation. Results. T(14;lS) wcs deteoted in 18 (72%)).37 (32%), 1 (4%) cod 2 (8%) pts from groups a, b, c, and d rcsptxtivcly (fl.01). All PBMC samples with t(14;lS) ovcrcxpressed b&2, these findings did not corrclctc with the prcscnc-ckbscnce of monoclonal Ifl component in the clyoprccipitatcs (MC type II/III), as well as with various cliiica-epidcmiological pwamctcrs; however, a trend toward a higher frequency of bcl-2 recanbination and ovcrcxpressioa was obscrvcd in tvw II MC (83%) es well cs in older ots and more.scvcre liver dkasc. In addition t($lS) wcs d&t& in 2/3 MC pts &b complicating malignant B-cell lymphoma Discussion. The significantly high frequency of t(14;18) in HCV-n&ted MC pts, suggests that HCV may play c role in the multistep benign end mcligoant lymphopmlifemtion. Tbe inhibition of B-cell apoptosis by bcl-2 oncogcnc should rcprcsent the early step of such pathogenctic pmccss. The detection of t(14;lS) also in polyclonal, type III MC and in c rclcvcnt proportion of patients with chronic HCV infection without MC further supports this hypothesis. Finally, the detection of t(14; 18) in HCV infected patients may be a predictive factor for the development of MC and/or other lymphoprolifcmtive disorders.