- Email: [email protected]
649 MOLECULAR EPIDEMIOLOGY OF HEPATITIS C VIRUS GENOTYPE 5A IN FRANCE (A MULTICENTRE STUDY ANRS)
POSTERS When HCVpp lacking the HVR were used (or when SRB1 was inhibited by BLT-4) serum enhancement was lost, particularly in HIV coinfected patients. In HIV patients, neutralization was positively and independently related to CD4 count (p < 0.01) and to antiretroviral therapy (p < 0.001). Selective pressure on HVR was higher in immunocompetent subjects and was related to the neutralization observed with serum immunoglobulin fraction. Conclusion: The sera of patients coinfected by HIV or liver transplanted are poorly neutralizing and facilitate the entry of HCVpp mainly through HVR and SRB1. Our findings are the first to show that low neutralization/high facilitation is related to the immune status and could be involved in the severity of HCV in immunocompromized subjects. 647 INSULIN DIRECTLY INHIBITS IFN-a SIGNALING IN HEPATIC CELLS THROUGH A SOCS3 INDEPENDENT PATHWAY L. Franceschini1 , M. Marcolongo1 , S. Mirandola1 , G. Bortoletto1 , A. Alberti2 , S. Realdon1 . 1 Venetian Institute of Molecular Medicine (VIMM), 2 Department of Histology, Microbiology and Medical Biotechnologies, University of Padua, Padova, Italy E-mail: [email protected] Background and Aims: Insulin resistance (IR) is one of the most important host factors associated with reduced response to PEGIFN plus ribavirin therapy in chronic hepatitis C (CHC). Intracellular factors, such as some members of SOCS family, associated with IR phenotype may play a role in the response to antiviral therapy as they are possibly involved in regulating IFN-a signaling. We therefore investigated “in vitro” the possible role of insulin in interfering with IFN-a pathway by analyzing gene and protein expression of some ISGs (PKR, MxA and OAS-1). Furthermore, to clarify the role of SOCS3 in cross-talking between insulin and IFN-a pathway we performed some experiments using siRNA technology. Methods: HepG2 were treated with different concentration of IFN-a and insulin alone or in combination. Time course analysis, of the expression of ISGs, was performed by Real Time PCR. PKR protein expression was also evaluated in the same settings by Westernblot analysis. Cells were then transfected with SiRNA for SOCS3 and gene expression of ISG were analyzed. Results: IFN-a alone enhanced the expression of ISGs. In HepG2 treated with IFN-a plus insulin a dose-dependent reduction in PKR, MxA and OAS-1 total mRNA levels was observed compared to untreated cells (p: 0.017, p: 0.103, p: 0.002 respectively with 100 nM insulin of stimuli; p: 0.0017, p: 0.0186, p: 0.006 respectively with 1000 nM insulin of stimuli). PKR protein expression showed the same trend of gene expression. Insulin alone had no effects on these genes and protein. Insulin mediated down-regulation of IFN-a antiviral activity was not mediated by over-expression of SOCS3 as shown in siRNA experiments. Conclusions: In HepG2 insulin significantly reduces the IFN-a mediated induction of three major ISGs, in a dose-dependent manner. Moreover SOCS3 over-expression induced by insulin does not represent the intracellular factor that can modulate the IFN-a cellular sensitivity. These experimental data indeed suggest that the control of insulin levels at baseline before initiation of IFN-a based therapy remains the best choice, in the clinical practice, to improve the antiviral response in CHC.
648 HEPATITIS C VIRUS CORE PROTEIN TRIGGERS HEPATIC ANGIOGENESIS BY A MECHANISM INCLUDING MULTIPLE PATHWAYS M. Hassan1 , H. Ghozlan2 , O. Abdel-Kader3 . 1 Lab. Mol. Tumour Therapy, Clinic of Dermatology, University Hospital of Duesseldorf, Duesseldorf, Germany; 2 Microbiology, Faculty of Science, Alexandria University, 3 Microbiology, Medical Research Institute, University of Alexandria, Alexandria, Egypt E-mail: [email protected] Background: Chronic hepatitis C virus (HCV) infection is associated with the production of serum cytokines, including transforming growth factor (TGF)-beta2. Despite the occurrence of hepatic angiogenesis in liver conditions, the role of HCV proteins in this context is currently unknown. Methods: ELISA, Western blot, Immune histochemistry, Immune fluorescence, in vitro kinase assay, electrophoretic mobility shift assay. Results: We demonstrated that the development of hepatic neoangiogenesis in patients infected with HCV is associated with the expression of TGF-beta2 and vascular endothelial growth factor (VEGF) and with activation of endothelial cells, as evidenced by CD34 expression. The analysis of liver biopsies of HCV-positive and HCV-negative patients using immunostaining showed significant elevation of TGF-beta2, VEGF, and CD34 expression in patients who were HCV-positive. We confirmed further the production of both TGF-beta2 and VEGF proteins, in the hepatoma cell lines infected with JFH1 or transfected with core. Furthermore, data obtained from inhibitor experiments revealed the importance of E2F1 and ASK1 in the modulation of core-induced activation of JNK and p38 pathways and suggested an essential role for JNK, p38, and ERK pathways in the regulation of core-induced production of TGFbeta2 and VEGF proteins. Conclusion: Our data provide insight into the molecular mechanisms whereby core protein mediates the development of hepatic angiogenesis in patients with chronic HCV infection (Hepatology. 2009 May; 49(5): 1469–82). 649 MOLECULAR EPIDEMIOLOGY OF HEPATITIS C VIRUS GENOTYPE 5A IN FRANCE (A MULTICENTRE STUDY ANRS) C. Henquell1 , J. Guglielmini2 , J. Verbeeck3 , J.-L. Bailly4 , R.-J. Perez Serra5 , M. Van Ranst3 , K. Randl6 , G. Bommelaer6 , H. Peigue-Lafeuille4 , A. Abergel6 , The ANRS HCV Group. 1 Virology, Clermont-Ferrand Teaching Hospital, Clermont-Ferrand, France; 2 Genetic and Physiology Bacterial Laboratory, University of Bruxelles, Gosselies, 3 Clinical and Epidemiological Virology, Rega Institute for Medical Research, University of Leuven, Leuven, Belgium; 4 EA3843-Virology, Faculty of Medicine, University of Auvergne, Clermont-Ferrand, France; 5 Digestive, Alcoy Hospital, Alicante, Spain; 6 Hepato-gastroenterology, Clermont-Ferrand Teaching Hospital, Clermont-Ferrand, France E-mail: [email protected] HCV genotype 5a (HCV 5a) was rare all over the world except in South Africa. In contrast to the other genotypes, a single subtype is described and its epidemic history is still poorly documented. To trace the origin of a local epidemic previously described in Central France (Abergel, APT, 2007) and to determine phylogenetic relationships among HCV 5a isolates originated from France, Spain, Belgium, South Africa and Algeria, we examined E1 and NS3/4 separated and concatenated genome regions. Phylogenetic analyses revealed a general clustering according to geographical origin. Using a Bayesian evolutionary method calibrated with known dates of proved transmissions by blood transfusion, we estimated the date of the most recent common ancestor (MRCA) of 97 HCV 5a sequences originated from Central France to 1953 (95% CIs: 1938–1963) and
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POSTERS confirmed the concomitant role of transfusion, iatrogenic route and intra familial transmission in HCV 5a local amplification. The same molecular dating method was applied to 120 diverse geographical isolates and allowed to estimate the date of divergence to 1895 (95% CIs: 1856–1922). The dates of the MRCA of South African, Spanish and Belgian clusters were estimated to 1929, 1941 and 1945 respectively. The similar diversity of geographical clusters and results of phylodynamic analysis suggested a concomitant spread from the end of the 19th century in the different countries via the transmission routes involved in Central France epidemic. 650 ANTIVIRAL ACTIVITY, COMBINATION AND RESISTANCE OF ACH1625, A POTENT, CLINICAL STAGE HCV NS3 PROTEASE INHIBITOR M. Huang, J. Fabrycki, D. Patel, S. Podos, G. Yang, Y. Zhao, C.W. Marlor, K. Stauber, A. Agarwal, X. Wang, M. Deshpande, A. Phadke. Achillion Pharmaceuticals, New Haven, CT, USA E-mail: [email protected] Background and Aims: HCV NS3 protease is a clinically validated target. Analysis of structural information of the target and its ligands led to design and synthesis of competitive reversible inhibitors. ACH-1625 demonstrates favorable pharmacokinetic and good safety profiles in animals. ACH-1625 is very well tolerated after administration of single and multiple doses up to 2000 mg in healthy volunteers. Currently, ACH-1625 is being evaluated in chronically-infected HCV patients. This report describes in vitro profiles of ACH-1625 including the activity against NS3 proteases, HCV replicons, host proteases and other viruses. The combination of ACH-1625 with other anti-HCV agents and its resistance profile is also presented. Methods: ACH-1625 was evaluated in biochemical assays against a panel of NS3 and cellular proteases. Inhibitory activity against HCV was also examined in a variety of HCV replicons. Combined effect of ACH-1625 with other inhibitors was studied in the replicon system. Resistant variants were selected after exposing genotype-1a or 1b HCV replicons to ACH-1625. Results: ACH-1625 exhibited high potency against HCV NS3 proteases (both genotyp1a and 1b) with IC50s <1 nM. Kinetic analyses showed that ACH-1625 bound the enzyme in two steps with the formation of a second highly stable inhibitor and enzyme complex. ACH-1625 potent activity was further confirmed against HCV replicons, including ones which contained NS3 protease domain from genotype 1a and 1b clinical isolates. In addition, ACH-1625 showed no activity against HIV-1, HSV-1, RSV, influenza and BVDV; and minimal cytotoxicity in a panel of cell lines and primary cells (CC50s >32 mM). ACH-1625 was specific for inhibition of NS3 protease; with no activity against 15/16 cellular proteases (IC50 >10 mM) and only weakly active against cathepsin S (IC50 = 4.5 mM). Combination of ACH-1625 with interferon, ribavirin and other experimental STAT-C agents yielded additive to synergistic effect. Finally, different mutations emerged from genotype-1a and 1b replicons under ACH-1625 selection and reverse genetics were conducted to investigate the role of these mutations. Conclusions: ACH-1625 possesses a potent and specific activity against HCV NS3 protease and acts additively or synergistically with other classes of inhibitors and warrants continued clinical development for treatment of hepatitis-C.
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651 GERANYLGERANYLACETONE INDUCED MTOR ACTIVITY HAS ANTI-HEPATITIS C VIRUS ACTIVITY T. Ichikawa, S. Takeshita, T. Muraoka, H. Matsuzaki, H. Miyaaki, N. Taura, K. Nakao. Department of Gastroenterology and Hepatology, Nagasaki Unversity Hospital, Nagasaki, Japan E-mail: [email protected] Background and Aims: New anti-hepatiti C virus (HCV) agents have been developed to inhibit the life cycle of HCV and are used in combination with type 1 interferon (IFN) to ameliorate the salvage rate of HCV infection. It is necessary to improve the salvage rate of HCV infection by clarifying the efficacy of IFN treatment since type 1 IFN is the most basic agent for HCV treatment. Geranylgeranylacetone (GGA), an isoprenoid compound, which includes retinoids, has been used orally as an anti-ulcer drug in Japan. GGA acts as a potent inducer of anti-viral gene expression by stimulating the IFN-stimulated gene factor 3 formation in human hepatoma cells (Ichikawa T., at. al., Biochemical and Biophysical Research Communications 2001; 280, 933–939). This study verified the anti-HCV activity of GGA in a replicon system. In addition, mechanisms of anti-HCV activity were examined in the replicon cells. Methods: OR6 cells stably harboring the full-length genotype 1 replicon containing the Renilla luciferase gene, ORN/C-5B/KE, were used to examine the influence of the anti-HCV effect of IFN. siRNA transfection was used for mTOR knockdown to explore role of mTOR in the anti-HCV activity. An mTOR activity assay was conducted to confirm the activity mechanism of GGA. Results: The results showed that GGA had the anti-HCV activity. GGA induced anti-HCV replicon activity in a time- and dosedependent manner. GGA did not activate the tyrosine 701 and serine 727 on STAT-1 and did not induce HSP-70 in OR6 cells. The anti-HCV effect depended on the GGA induced mTOR activity, not STAT-1 activity and PKR. An additive effect was observed with a combination of IFN and GGA. Conclusion: GGA, a drug that can be safely administered orally, has mTOR dependent anti-HCV activity. The combination of IFN and GGA has additive effect on anti-HCV activity. There is a possibility that the GGA anti-HCV activity can be complimented by IFN. It will be necessary to examine the clinical effectiveness of the combination of GGA and IFN for HCV patients in the future. 652 RECOMBINANT MAMMALIAN CELL DERIVED HEPATITIS C VIRUS LIKE PARTICLES INDUCE NEUTRALISING ANTIBODY AND B CELL RESPONSES TO HEPATITIS C VIRUS D.F. Johnson1,2 , B. Chua3 , R. Chin2 , L. Earnest-Silveira2 , H. Zentgraf4 , C.-T. Bock5 , W. Zeng3 , D.C. Jacskon3 , J. Torresi1,2 . 1 Department of Infectious Diseases, Austin Hospital, 2 Department of Medicine, Austin Hospital, University of Melbourne, 3 Department of Microbiology and Immunology, The University of Melbourne, Melbourne, VIC, Australia; 4 Applied Tumor Virology, German Cancer Research Center (DKFZ), Heidelberg, 5 Department Molecular Pathology, University Hospital of Tuebingen, Tuebingen, Germany E-mail: [email protected] Background and Aims: Clearance of Hepatitis C virus (HCV) requires a strong and broadly cross-reactive CD4+, CD8+ T cell and neutralising antibody (Ab) responses. Virus like particles (VLPs) resemble mature parent virus inducing protective humoral and cellular immune responses against HCV and provide a viable prophylactic vaccine candidate. Methods: Recombinant adenoviruses expressing HCV genotype 1a core-E1-E2 were used to infect Huh7 (hepatoma) cells. HCV VLPs were isolated and purified by CsCl density gradient ultracentrifugation. HCV VLPs were used to test maturation of mouse bone marrow derived Dendritic cells (DC). Mice were
Journal of Hepatology 2010 vol. 52 | S183–S317
648 HEPATITIS C VIRUS CORE PROTEIN TRIGGERS HEPATIC ANGIOGENESIS BY A MECHANISM INCLUDING MULTIPLE PATHWAYS M. Hassan1 , H. Ghozlan2 , O. Abdel-Kader3 . 1 Lab. Mol. Tumour Therapy, Clinic of Dermatology, University Hospital of Duesseldorf, Duesseldorf, Germany; 2 Microbiology, Faculty of Science, Alexandria University, 3 Microbiology, Medical Research Institute, University of Alexandria, Alexandria, Egypt E-mail: [email protected] Background: Chronic hepatitis C virus (HCV) infection is associated with the production of serum cytokines, including transforming growth factor (TGF)-beta2. Despite the occurrence of hepatic angiogenesis in liver conditions, the role of HCV proteins in this context is currently unknown. Methods: ELISA, Western blot, Immune histochemistry, Immune fluorescence, in vitro kinase assay, electrophoretic mobility shift assay. Results: We demonstrated that the development of hepatic neoangiogenesis in patients infected with HCV is associated with the expression of TGF-beta2 and vascular endothelial growth factor (VEGF) and with activation of endothelial cells, as evidenced by CD34 expression. The analysis of liver biopsies of HCV-positive and HCV-negative patients using immunostaining showed significant elevation of TGF-beta2, VEGF, and CD34 expression in patients who were HCV-positive. We confirmed further the production of both TGF-beta2 and VEGF proteins, in the hepatoma cell lines infected with JFH1 or transfected with core. Furthermore, data obtained from inhibitor experiments revealed the importance of E2F1 and ASK1 in the modulation of core-induced activation of JNK and p38 pathways and suggested an essential role for JNK, p38, and ERK pathways in the regulation of core-induced production of TGFbeta2 and VEGF proteins. Conclusion: Our data provide insight into the molecular mechanisms whereby core protein mediates the development of hepatic angiogenesis in patients with chronic HCV infection (Hepatology. 2009 May; 49(5): 1469–82). 649 MOLECULAR EPIDEMIOLOGY OF HEPATITIS C VIRUS GENOTYPE 5A IN FRANCE (A MULTICENTRE STUDY ANRS) C. Henquell1 , J. Guglielmini2 , J. Verbeeck3 , J.-L. Bailly4 , R.-J. Perez Serra5 , M. Van Ranst3 , K. Randl6 , G. Bommelaer6 , H. Peigue-Lafeuille4 , A. Abergel6 , The ANRS HCV Group. 1 Virology, Clermont-Ferrand Teaching Hospital, Clermont-Ferrand, France; 2 Genetic and Physiology Bacterial Laboratory, University of Bruxelles, Gosselies, 3 Clinical and Epidemiological Virology, Rega Institute for Medical Research, University of Leuven, Leuven, Belgium; 4 EA3843-Virology, Faculty of Medicine, University of Auvergne, Clermont-Ferrand, France; 5 Digestive, Alcoy Hospital, Alicante, Spain; 6 Hepato-gastroenterology, Clermont-Ferrand Teaching Hospital, Clermont-Ferrand, France E-mail: [email protected] HCV genotype 5a (HCV 5a) was rare all over the world except in South Africa. In contrast to the other genotypes, a single subtype is described and its epidemic history is still poorly documented. To trace the origin of a local epidemic previously described in Central France (Abergel, APT, 2007) and to determine phylogenetic relationships among HCV 5a isolates originated from France, Spain, Belgium, South Africa and Algeria, we examined E1 and NS3/4 separated and concatenated genome regions. Phylogenetic analyses revealed a general clustering according to geographical origin. Using a Bayesian evolutionary method calibrated with known dates of proved transmissions by blood transfusion, we estimated the date of the most recent common ancestor (MRCA) of 97 HCV 5a sequences originated from Central France to 1953 (95% CIs: 1938–1963) and
Journal of Hepatology 2010 vol. 52 | S183–S317
S253
POSTERS confirmed the concomitant role of transfusion, iatrogenic route and intra familial transmission in HCV 5a local amplification. The same molecular dating method was applied to 120 diverse geographical isolates and allowed to estimate the date of divergence to 1895 (95% CIs: 1856–1922). The dates of the MRCA of South African, Spanish and Belgian clusters were estimated to 1929, 1941 and 1945 respectively. The similar diversity of geographical clusters and results of phylodynamic analysis suggested a concomitant spread from the end of the 19th century in the different countries via the transmission routes involved in Central France epidemic. 650 ANTIVIRAL ACTIVITY, COMBINATION AND RESISTANCE OF ACH1625, A POTENT, CLINICAL STAGE HCV NS3 PROTEASE INHIBITOR M. Huang, J. Fabrycki, D. Patel, S. Podos, G. Yang, Y. Zhao, C.W. Marlor, K. Stauber, A. Agarwal, X. Wang, M. Deshpande, A. Phadke. Achillion Pharmaceuticals, New Haven, CT, USA E-mail: [email protected] Background and Aims: HCV NS3 protease is a clinically validated target. Analysis of structural information of the target and its ligands led to design and synthesis of competitive reversible inhibitors. ACH-1625 demonstrates favorable pharmacokinetic and good safety profiles in animals. ACH-1625 is very well tolerated after administration of single and multiple doses up to 2000 mg in healthy volunteers. Currently, ACH-1625 is being evaluated in chronically-infected HCV patients. This report describes in vitro profiles of ACH-1625 including the activity against NS3 proteases, HCV replicons, host proteases and other viruses. The combination of ACH-1625 with other anti-HCV agents and its resistance profile is also presented. Methods: ACH-1625 was evaluated in biochemical assays against a panel of NS3 and cellular proteases. Inhibitory activity against HCV was also examined in a variety of HCV replicons. Combined effect of ACH-1625 with other inhibitors was studied in the replicon system. Resistant variants were selected after exposing genotype-1a or 1b HCV replicons to ACH-1625. Results: ACH-1625 exhibited high potency against HCV NS3 proteases (both genotyp1a and 1b) with IC50s <1 nM. Kinetic analyses showed that ACH-1625 bound the enzyme in two steps with the formation of a second highly stable inhibitor and enzyme complex. ACH-1625 potent activity was further confirmed against HCV replicons, including ones which contained NS3 protease domain from genotype 1a and 1b clinical isolates. In addition, ACH-1625 showed no activity against HIV-1, HSV-1, RSV, influenza and BVDV; and minimal cytotoxicity in a panel of cell lines and primary cells (CC50s >32 mM). ACH-1625 was specific for inhibition of NS3 protease; with no activity against 15/16 cellular proteases (IC50 >10 mM) and only weakly active against cathepsin S (IC50 = 4.5 mM). Combination of ACH-1625 with interferon, ribavirin and other experimental STAT-C agents yielded additive to synergistic effect. Finally, different mutations emerged from genotype-1a and 1b replicons under ACH-1625 selection and reverse genetics were conducted to investigate the role of these mutations. Conclusions: ACH-1625 possesses a potent and specific activity against HCV NS3 protease and acts additively or synergistically with other classes of inhibitors and warrants continued clinical development for treatment of hepatitis-C.
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651 GERANYLGERANYLACETONE INDUCED MTOR ACTIVITY HAS ANTI-HEPATITIS C VIRUS ACTIVITY T. Ichikawa, S. Takeshita, T. Muraoka, H. Matsuzaki, H. Miyaaki, N. Taura, K. Nakao. Department of Gastroenterology and Hepatology, Nagasaki Unversity Hospital, Nagasaki, Japan E-mail: [email protected] Background and Aims: New anti-hepatiti C virus (HCV) agents have been developed to inhibit the life cycle of HCV and are used in combination with type 1 interferon (IFN) to ameliorate the salvage rate of HCV infection. It is necessary to improve the salvage rate of HCV infection by clarifying the efficacy of IFN treatment since type 1 IFN is the most basic agent for HCV treatment. Geranylgeranylacetone (GGA), an isoprenoid compound, which includes retinoids, has been used orally as an anti-ulcer drug in Japan. GGA acts as a potent inducer of anti-viral gene expression by stimulating the IFN-stimulated gene factor 3 formation in human hepatoma cells (Ichikawa T., at. al., Biochemical and Biophysical Research Communications 2001; 280, 933–939). This study verified the anti-HCV activity of GGA in a replicon system. In addition, mechanisms of anti-HCV activity were examined in the replicon cells. Methods: OR6 cells stably harboring the full-length genotype 1 replicon containing the Renilla luciferase gene, ORN/C-5B/KE, were used to examine the influence of the anti-HCV effect of IFN. siRNA transfection was used for mTOR knockdown to explore role of mTOR in the anti-HCV activity. An mTOR activity assay was conducted to confirm the activity mechanism of GGA. Results: The results showed that GGA had the anti-HCV activity. GGA induced anti-HCV replicon activity in a time- and dosedependent manner. GGA did not activate the tyrosine 701 and serine 727 on STAT-1 and did not induce HSP-70 in OR6 cells. The anti-HCV effect depended on the GGA induced mTOR activity, not STAT-1 activity and PKR. An additive effect was observed with a combination of IFN and GGA. Conclusion: GGA, a drug that can be safely administered orally, has mTOR dependent anti-HCV activity. The combination of IFN and GGA has additive effect on anti-HCV activity. There is a possibility that the GGA anti-HCV activity can be complimented by IFN. It will be necessary to examine the clinical effectiveness of the combination of GGA and IFN for HCV patients in the future. 652 RECOMBINANT MAMMALIAN CELL DERIVED HEPATITIS C VIRUS LIKE PARTICLES INDUCE NEUTRALISING ANTIBODY AND B CELL RESPONSES TO HEPATITIS C VIRUS D.F. Johnson1,2 , B. Chua3 , R. Chin2 , L. Earnest-Silveira2 , H. Zentgraf4 , C.-T. Bock5 , W. Zeng3 , D.C. Jacskon3 , J. Torresi1,2 . 1 Department of Infectious Diseases, Austin Hospital, 2 Department of Medicine, Austin Hospital, University of Melbourne, 3 Department of Microbiology and Immunology, The University of Melbourne, Melbourne, VIC, Australia; 4 Applied Tumor Virology, German Cancer Research Center (DKFZ), Heidelberg, 5 Department Molecular Pathology, University Hospital of Tuebingen, Tuebingen, Germany E-mail: [email protected] Background and Aims: Clearance of Hepatitis C virus (HCV) requires a strong and broadly cross-reactive CD4+, CD8+ T cell and neutralising antibody (Ab) responses. Virus like particles (VLPs) resemble mature parent virus inducing protective humoral and cellular immune responses against HCV and provide a viable prophylactic vaccine candidate. Methods: Recombinant adenoviruses expressing HCV genotype 1a core-E1-E2 were used to infect Huh7 (hepatoma) cells. HCV VLPs were isolated and purified by CsCl density gradient ultracentrifugation. HCV VLPs were used to test maturation of mouse bone marrow derived Dendritic cells (DC). Mice were
Journal of Hepatology 2010 vol. 52 | S183–S317