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Host genetic factors and the natural course of disease in chronic hepatitis C infection
HEPATOLOGYVol. 34, No. 4, Ft. 2, 2001
AASLD ABSTRACTS
223A
197
198
HOST GENETIC FACTORS A N D THE NATURAL COURSE OF DISEASE IN CHRONIC HEPATITIS C INFECTION. Holger M Hinriehsen, Silvia Mascheretti, Susanne Ross, Christian-Albrechts-University, K i d Germany; Peter Buggisch, Universitatsklinikum Eppendorff, H a m b u r g Germany; Ulrich R Foelsch, Stefan Schreiber, Christian-Albrechts-University, K i d Germany
PERFORMANCE OF THE N E W BAYER VERSANT TM HCV 3.0 RNA ASSAY (BDNA) FOR Q U A N T I T A T I O N OF HCV RNA IN PLASMA A N D SERUM: CONVERSION TO INTERNATIONAL UNITS A N D COMPARISON W I T H THE ROCHE COBAS AMPLICOR HCV M O N I T O R 2.0 ASSAY. Marcel BeId, Roel Sentjens, Sjoerd Rebers, Christine J Weegink, Jan Weel, Cees Sol, Rene Boom, Acad Medical Ctr, Amsterdam Netherlands
Background: Chronic hepatitis occurred in the majority of Hepatitis C infected patients. The natural course of chronic liver disease ranged from a mild asymptomatic infection without liver fibrosis up to severe liver disease with cirrhosis and hepatocellular cancer. Several factors have been reported to influence the natural course of the disease, but there are still questions unresolved. Aim: We were therefore interested to investigate host genetic factors for their influence in the natural course of hepatitis C infection. Patients: 346 HCV infected patients from the universities of Kid and H a m b u r g were enrolled. Furthermore 376 unrelated sex and age matched healthy controls were investigated. Methods: More then 25 single nucleotide polymorhisms (SNPs)in genes related to cytokine activity were determined. These includes SNPs in the TNF alpha and beta genes, TNF receptor 1 and 2 genes, Interleukin 1 alpha and beta genes as well in the chemokine receptor 2 and 5 genes. SNPS were anlysed by multivariate testing for fibrosis progression, spontaneous recovery and chronicity. Results: In almost all of the chronic infected patients a liver biopsy was performed. In a significant percentage of patients HCV infection of more than 10 or 15 years was documented. There were no associations found for fibrosis progression and specific SNP findings. Only for the rare allele fie at aa position 64 in the CCR 2 gene an assocition with the inability to spontaneous recovery could be seen. Associations of SNP findings and reponse to interferon based therapy will be presented furthermore. Conclusion: There are up to now no k n o w n mutations in host genetic genes that are influencing fibrosis progression in chronic hepatitis C. There are first hints for a CCR 2 gene polymorhism to influence the chance of spontaneous recovery in HCV infected patients.
We have evaluated the VERSANTHCV 3.0 assay (bDNA) (VERSANTTM, Bayer Diagnostics, Berkeley, CA), which is an improved signal amplification procedure of the HCV 2.0 bDNA assay for the quantitation of HCV RNA in serum or plasma of HCV infected individuals. The VERSANTHCV 3.0 assay has a linear dynamic range of 2.5 x 103 to 4.0 x 107 HCV RNA copies per ml (c/ml). The performance of the VERSANT HCV 3.0 assay was evaluated using 3 different test panels. (i) Seronegativepanel (96 HCV negative specimens) tested in single well determinations in 4 independent runs (48 specimens per ran); (ii)A six member Dilution Panel consisting of HCV-3a virus with HCV RNA loads expressed in c/ml and tested in replicates of six in 2 independent runs; (iii) A six member HCV RNA quantitation panel (NAPTM) consisting of HCV-lb virus with HC¥ RNA loads expressed in International Units (IU/ml) and tested in replicates of six in 2 independent runs. The NAPTM panel was also used to calculate the factor for conversion of c/ml to IU/ml. To determine the quantitative correlation in IU/ml between the VERSANTHCV 3.0 assay and the HCV Monitor 2.0 assay (COBAS AMPLICORTM, Roche Diagnostic Systems, Branchburg, NJ), 102 patient samples were quantified by both assays and stratified according to the dynamic range and the thresholds of both assays. An overall specificity of 96.8% relative to the detection limit of the VERSANTHCV 3.0 assay was found. The intra and inter run reproducibility for both the Dilution Panel and NAPTM panel was consistent with coefficients of variation of less than 9%. Quautitation with the VERSANTHCV 3.0 assay was linear over the entire range of both panels (range of 4.4 x 103 to 3.5 x 106 c/ml and 5 x 103 to 2 x 106 IU/ml respectively) with correlation coefficients of 0.999, and slopes close to one and intercepts close to zero. The regression equation indicated that I IU corresponded to about 4.8 copies of HCV RNA. A correlation coefficient of 0.941 was found for HCV RNA values (in IU/ml) from the VERSANT HCV 3.0 and the HCV Monitor 2.0 assays; the regression line characterizing the values from the two assays exhibited a slope of 0.765 and an intercept of 1.109. It appeared that quantification values obtained from the HCV Monitor 2.0 assay between 5 x 102 and 1051U/ml were in general higher than those from the VERSANTHCV 3.0 assay, whereas values from the HCV Monitor 2.0 assay were underestimated for samples with HCV RNA levels above 10~ IU/ml. Quantitative results obtained close to the lower limit of the VERSANTHCV 3.0 assay might imply that its lower limit should be reconsidered. In summary, the VERSANTHCV 3.0assay allows accurate quantitation of HCV RNA levels expressed in IU/ml in plasma with a wide dynamic range (5.2 x 102 to 8.3 x 106 IU/ml corresponding to 2.5 x 10~to 4.0 x 107 copies ofHCV RNA/ml)and will be clinically relevant in the decision of duration of therapy and in tailoring during therapy for patients chronically infected with HCV.
199
200
COMPARING THE PUBLIC HEALTH BURDEN OF CHRONIC HEPATITIS C A N D HIV INFECTION IN FRANCE. Sylvie Deuffic-Burban, lnstitut Pasteur de Lille - INSERM U547, Lille France; John B Wong, New England Medical Center, Boston, MA; Thierry Poynard, Service d'Hepato Gastroenterologie, Groupe Hospitalier Pitie-Salpetriere, Paris France
INFECTION WITH MULTIPLE HEPATITIS VIRUSES: EVIDENCE FOR SUPPRESSION OF HCV REPLICATION BY HDV A N D HBV. Heiner Wedemeyer, Hans L Ttllmann, BjOrn Tegtmeyer, Markus Cornberg, Andreas Sch,aler, Heike Liermann, Christian Trautwein, Michael P Manns, Dept of Gastroneterology, Hannover Medical School, Hannover Germany
Infection wi h the bepa s C v rus (HCV) is more common than that ,ruth HIV, but HIV infection confers a poorer prognosis. PURPOSE OF STUDY:to compare the disease burden (incidence and mortality) related to HCV and to HIV. METHODS: Using mortality data and natural history esamates, we applied the backcalculatton method to make enceandmortalitv fromHCVandHIVunti11997. TheHCVmodela pliesnaturalhistorydata see Table) and hepatocellular carcinmna (HCC) mor ality da a from na ona s a ist cs. ~ e HIV model uses AIDS cases reported to the French National Instisute for Public Health Surveillance and HIV/AIDSdeaths reported to the French Institute of Health and lvledical Research It assumes median AIDS incubation times of 8 years in the absence of treatment or during mono-therapy period, 10 years during bi-therapy period and 20 years during tri-therapy period according to estimates from HtV seropositive subjects in French hospitals RESULTS:HCV incidence is estimated to be maximal in I989 with 32,800 infections [30,100;36,200] and about 20,900 [19,000;23,300] in 1997. HIV incidence is estimated to be maximal in 1987 with 9.000 infectious [7,700; 10.300] and 700 1500;900] in 1997. Related mortality is estimated to be about 2,900 [2,100;+,500] in 1997 for HCV compared to 1 300 for HIV. CONCLUSION: The public hea th burden of HCV ts on the rise, while the burden of HIV, given the fairly widespread use of effective medications, may be on the decline.
The impact of HBV and HDV co-infection on chronic hepatitis C virus infection had been a matter of debate. While the clinical course of chronic hepatitis C is thought to be worsened by HBV and HDV, it has been suggested that HBV suppresses replication of HCV. We investigated serological data of 3170 consecutive anti-HCV positive patients collected between 1992 and 2000 at the Hannover Medical School, Germany. Liver biopsies were available in a subgroup of 303 patients. Results: 170 (5.2%) individuals were co-infected with HCV and HBV (HBsAg+ / anti-HCVRNA+). HCV-RNA was detected by PCR (detection limit 600 IU/ml) in 82% of the 1163 patients who had recovered from HBV (anti-HBc-positive), in 85 % of the 1837 patients with no apparent previous contact to HBV (anti-HBc-negative) but only in 44% of the HBsAg+/anti-HCV+ patients (p<0.0001) indicating suppression of HCV replication by active HBV co-infection. Antibodies against hepatitis D virus were found in 33% of HBsAg+/anti-HCV+ patients. As expected, anti-HDV-antibodies tested more often positive in patients with low HBV-replication (HBsAg+/ HBV-PCR-; 38% anti-HDV-positive) than in patients with high HBV replication (HBV-DNA+, 18% anti-HDV-positive). Interestingly, similar results were obtained regarding HCV replication in HBV/I-ICV-coinfected patients (46% anti-HDV-positive in HCV-RNA- patients, vs. 13% in HCV-RNA+ patients; p<0.0001). Prevalence of anti-HDV antibodies in the different subgroups of HBV/HCV co-infected patients are shown in the table. The cIinical courses of co-infected patients varied widely. Overall fibrosis scores were higher in coinfected patients than in HBsAg-negative individuals (Ishak score: 3.7 vs. 2.3, p<0.001), which might he explained by a different risk-factor profile of patients infected with multiple viruses (history of intravenous drug abuse in 45% of the patients). However, several patients presented with very mild liver disease despite being co-infected with HCV and HBV. Surprisingly, inflammatory activities were lower in patients with active HCV replication than in HCV-RNA-negative individuals (p<0.02). CONCLUSIONS: HBV co-infection as well as HDV infection is associated with decreased HCV-RNA replication. However, lower histological disease activity in HCV viremic patients suggest a beneficial effect of HCV replication on the course of some HBsAg-positive patients. Treatment guidelines for patients with multiple hepatitis virus infections have to be developed.
~rojecaonsabouttnci
Pa~ar~etersof #~eHCVmodel Parameters proportion of new i~ltectloi~s that progressto chror=tettyo~e year ~ e r iathctioi~ Transition prob~bilgy(per year}from c~ro~ic hep~tet to cirrhosis (~e and sex dependent) Transi~n preb~btllty(per ye~) from cirthes[s to deathfrom liver failure {age,dep~detR) Ttensifio~ probability ~ year}from cirrhosis to NCC (age. depe~t) Transition probability{ba~year} from HCC to death from HCC Pmpertion of HCC de~bs attdbutetdeto HCC Prevalence of HCV chronic c~triers in 1994~nFrance Propo~oti of {rooted ~ubjec~ amotlgchro~licca~le~s Propozllo~of recovering among tre~todsubjects
Model values 7~% estimat~Jby me process 0928× I 04(~e~ ) 0028,1 05e'~s~l 0,33 2D~ 590:0~0 5% ~rct~1991 to 1994 I0% ~ m 1995',o ! 9 ~ 19% flora 1991 ~o1994 20% f~om19~ D 1997
JS~
S~aO
I
Prevalence of anti-HDV-antibodies in 129 HBsAg+lanti-HCV+ individuals I-IBV-DNA-positive HBV-DNA-negative HCV-RNA-positive 0/9 (0%) 6t39 (15%) HOV-RNA-negative 8/22 (27%) 26/45 (58%)
AASLD ABSTRACTS
223A
197
198
HOST GENETIC FACTORS A N D THE NATURAL COURSE OF DISEASE IN CHRONIC HEPATITIS C INFECTION. Holger M Hinriehsen, Silvia Mascheretti, Susanne Ross, Christian-Albrechts-University, K i d Germany; Peter Buggisch, Universitatsklinikum Eppendorff, H a m b u r g Germany; Ulrich R Foelsch, Stefan Schreiber, Christian-Albrechts-University, K i d Germany
PERFORMANCE OF THE N E W BAYER VERSANT TM HCV 3.0 RNA ASSAY (BDNA) FOR Q U A N T I T A T I O N OF HCV RNA IN PLASMA A N D SERUM: CONVERSION TO INTERNATIONAL UNITS A N D COMPARISON W I T H THE ROCHE COBAS AMPLICOR HCV M O N I T O R 2.0 ASSAY. Marcel BeId, Roel Sentjens, Sjoerd Rebers, Christine J Weegink, Jan Weel, Cees Sol, Rene Boom, Acad Medical Ctr, Amsterdam Netherlands
Background: Chronic hepatitis occurred in the majority of Hepatitis C infected patients. The natural course of chronic liver disease ranged from a mild asymptomatic infection without liver fibrosis up to severe liver disease with cirrhosis and hepatocellular cancer. Several factors have been reported to influence the natural course of the disease, but there are still questions unresolved. Aim: We were therefore interested to investigate host genetic factors for their influence in the natural course of hepatitis C infection. Patients: 346 HCV infected patients from the universities of Kid and H a m b u r g were enrolled. Furthermore 376 unrelated sex and age matched healthy controls were investigated. Methods: More then 25 single nucleotide polymorhisms (SNPs)in genes related to cytokine activity were determined. These includes SNPs in the TNF alpha and beta genes, TNF receptor 1 and 2 genes, Interleukin 1 alpha and beta genes as well in the chemokine receptor 2 and 5 genes. SNPS were anlysed by multivariate testing for fibrosis progression, spontaneous recovery and chronicity. Results: In almost all of the chronic infected patients a liver biopsy was performed. In a significant percentage of patients HCV infection of more than 10 or 15 years was documented. There were no associations found for fibrosis progression and specific SNP findings. Only for the rare allele fie at aa position 64 in the CCR 2 gene an assocition with the inability to spontaneous recovery could be seen. Associations of SNP findings and reponse to interferon based therapy will be presented furthermore. Conclusion: There are up to now no k n o w n mutations in host genetic genes that are influencing fibrosis progression in chronic hepatitis C. There are first hints for a CCR 2 gene polymorhism to influence the chance of spontaneous recovery in HCV infected patients.
We have evaluated the VERSANTHCV 3.0 assay (bDNA) (VERSANTTM, Bayer Diagnostics, Berkeley, CA), which is an improved signal amplification procedure of the HCV 2.0 bDNA assay for the quantitation of HCV RNA in serum or plasma of HCV infected individuals. The VERSANTHCV 3.0 assay has a linear dynamic range of 2.5 x 103 to 4.0 x 107 HCV RNA copies per ml (c/ml). The performance of the VERSANT HCV 3.0 assay was evaluated using 3 different test panels. (i) Seronegativepanel (96 HCV negative specimens) tested in single well determinations in 4 independent runs (48 specimens per ran); (ii)A six member Dilution Panel consisting of HCV-3a virus with HCV RNA loads expressed in c/ml and tested in replicates of six in 2 independent runs; (iii) A six member HCV RNA quantitation panel (NAPTM) consisting of HCV-lb virus with HC¥ RNA loads expressed in International Units (IU/ml) and tested in replicates of six in 2 independent runs. The NAPTM panel was also used to calculate the factor for conversion of c/ml to IU/ml. To determine the quantitative correlation in IU/ml between the VERSANTHCV 3.0 assay and the HCV Monitor 2.0 assay (COBAS AMPLICORTM, Roche Diagnostic Systems, Branchburg, NJ), 102 patient samples were quantified by both assays and stratified according to the dynamic range and the thresholds of both assays. An overall specificity of 96.8% relative to the detection limit of the VERSANTHCV 3.0 assay was found. The intra and inter run reproducibility for both the Dilution Panel and NAPTM panel was consistent with coefficients of variation of less than 9%. Quautitation with the VERSANTHCV 3.0 assay was linear over the entire range of both panels (range of 4.4 x 103 to 3.5 x 106 c/ml and 5 x 103 to 2 x 106 IU/ml respectively) with correlation coefficients of 0.999, and slopes close to one and intercepts close to zero. The regression equation indicated that I IU corresponded to about 4.8 copies of HCV RNA. A correlation coefficient of 0.941 was found for HCV RNA values (in IU/ml) from the VERSANT HCV 3.0 and the HCV Monitor 2.0 assays; the regression line characterizing the values from the two assays exhibited a slope of 0.765 and an intercept of 1.109. It appeared that quantification values obtained from the HCV Monitor 2.0 assay between 5 x 102 and 1051U/ml were in general higher than those from the VERSANTHCV 3.0 assay, whereas values from the HCV Monitor 2.0 assay were underestimated for samples with HCV RNA levels above 10~ IU/ml. Quantitative results obtained close to the lower limit of the VERSANTHCV 3.0 assay might imply that its lower limit should be reconsidered. In summary, the VERSANTHCV 3.0assay allows accurate quantitation of HCV RNA levels expressed in IU/ml in plasma with a wide dynamic range (5.2 x 102 to 8.3 x 106 IU/ml corresponding to 2.5 x 10~to 4.0 x 107 copies ofHCV RNA/ml)and will be clinically relevant in the decision of duration of therapy and in tailoring during therapy for patients chronically infected with HCV.
199
200
COMPARING THE PUBLIC HEALTH BURDEN OF CHRONIC HEPATITIS C A N D HIV INFECTION IN FRANCE. Sylvie Deuffic-Burban, lnstitut Pasteur de Lille - INSERM U547, Lille France; John B Wong, New England Medical Center, Boston, MA; Thierry Poynard, Service d'Hepato Gastroenterologie, Groupe Hospitalier Pitie-Salpetriere, Paris France
INFECTION WITH MULTIPLE HEPATITIS VIRUSES: EVIDENCE FOR SUPPRESSION OF HCV REPLICATION BY HDV A N D HBV. Heiner Wedemeyer, Hans L Ttllmann, BjOrn Tegtmeyer, Markus Cornberg, Andreas Sch,aler, Heike Liermann, Christian Trautwein, Michael P Manns, Dept of Gastroneterology, Hannover Medical School, Hannover Germany
Infection wi h the bepa s C v rus (HCV) is more common than that ,ruth HIV, but HIV infection confers a poorer prognosis. PURPOSE OF STUDY:to compare the disease burden (incidence and mortality) related to HCV and to HIV. METHODS: Using mortality data and natural history esamates, we applied the backcalculatton method to make enceandmortalitv fromHCVandHIVunti11997. TheHCVmodela pliesnaturalhistorydata see Table) and hepatocellular carcinmna (HCC) mor ality da a from na ona s a ist cs. ~ e HIV model uses AIDS cases reported to the French National Instisute for Public Health Surveillance and HIV/AIDSdeaths reported to the French Institute of Health and lvledical Research It assumes median AIDS incubation times of 8 years in the absence of treatment or during mono-therapy period, 10 years during bi-therapy period and 20 years during tri-therapy period according to estimates from HtV seropositive subjects in French hospitals RESULTS:HCV incidence is estimated to be maximal in I989 with 32,800 infections [30,100;36,200] and about 20,900 [19,000;23,300] in 1997. HIV incidence is estimated to be maximal in 1987 with 9.000 infectious [7,700; 10.300] and 700 1500;900] in 1997. Related mortality is estimated to be about 2,900 [2,100;+,500] in 1997 for HCV compared to 1 300 for HIV. CONCLUSION: The public hea th burden of HCV ts on the rise, while the burden of HIV, given the fairly widespread use of effective medications, may be on the decline.
The impact of HBV and HDV co-infection on chronic hepatitis C virus infection had been a matter of debate. While the clinical course of chronic hepatitis C is thought to be worsened by HBV and HDV, it has been suggested that HBV suppresses replication of HCV. We investigated serological data of 3170 consecutive anti-HCV positive patients collected between 1992 and 2000 at the Hannover Medical School, Germany. Liver biopsies were available in a subgroup of 303 patients. Results: 170 (5.2%) individuals were co-infected with HCV and HBV (HBsAg+ / anti-HCVRNA+). HCV-RNA was detected by PCR (detection limit 600 IU/ml) in 82% of the 1163 patients who had recovered from HBV (anti-HBc-positive), in 85 % of the 1837 patients with no apparent previous contact to HBV (anti-HBc-negative) but only in 44% of the HBsAg+/anti-HCV+ patients (p<0.0001) indicating suppression of HCV replication by active HBV co-infection. Antibodies against hepatitis D virus were found in 33% of HBsAg+/anti-HCV+ patients. As expected, anti-HDV-antibodies tested more often positive in patients with low HBV-replication (HBsAg+/ HBV-PCR-; 38% anti-HDV-positive) than in patients with high HBV replication (HBV-DNA+, 18% anti-HDV-positive). Interestingly, similar results were obtained regarding HCV replication in HBV/I-ICV-coinfected patients (46% anti-HDV-positive in HCV-RNA- patients, vs. 13% in HCV-RNA+ patients; p<0.0001). Prevalence of anti-HDV antibodies in the different subgroups of HBV/HCV co-infected patients are shown in the table. The cIinical courses of co-infected patients varied widely. Overall fibrosis scores were higher in coinfected patients than in HBsAg-negative individuals (Ishak score: 3.7 vs. 2.3, p<0.001), which might he explained by a different risk-factor profile of patients infected with multiple viruses (history of intravenous drug abuse in 45% of the patients). However, several patients presented with very mild liver disease despite being co-infected with HCV and HBV. Surprisingly, inflammatory activities were lower in patients with active HCV replication than in HCV-RNA-negative individuals (p<0.02). CONCLUSIONS: HBV co-infection as well as HDV infection is associated with decreased HCV-RNA replication. However, lower histological disease activity in HCV viremic patients suggest a beneficial effect of HCV replication on the course of some HBsAg-positive patients. Treatment guidelines for patients with multiple hepatitis virus infections have to be developed.
~rojecaonsabouttnci
Pa~ar~etersof #~eHCVmodel Parameters proportion of new i~ltectloi~s that progressto chror=tettyo~e year ~ e r iathctioi~ Transition prob~bilgy(per year}from c~ro~ic hep~tet to cirrhosis (~e and sex dependent) Transi~n preb~btllty(per ye~) from cirthes[s to deathfrom liver failure {age,dep~detR) Ttensifio~ probability ~ year}from cirrhosis to NCC (age. depe~t) Transition probability{ba~year} from HCC to death from HCC Pmpertion of HCC de~bs attdbutetdeto HCC Prevalence of HCV chronic c~triers in 1994~nFrance Propo~oti of {rooted ~ubjec~ amotlgchro~licca~le~s Propozllo~of recovering among tre~todsubjects
Model values 7~% estimat~Jby me process 0928× I 04(~e~ ) 0028,1 05e'~s~l 0,33 2D~ 590:0~0 5% ~rct~1991 to 1994 I0% ~ m 1995',o ! 9 ~ 19% flora 1991 ~o1994 20% f~om19~ D 1997
JS~
S~aO
I
Prevalence of anti-HDV-antibodies in 129 HBsAg+lanti-HCV+ individuals I-IBV-DNA-positive HBV-DNA-negative HCV-RNA-positive 0/9 (0%) 6t39 (15%) HOV-RNA-negative 8/22 (27%) 26/45 (58%)