5 years (range: 6.0 – 11.9 years). At baseline, 13 patients had 100mg lamivudine, 11 had 600mg telbivudine, 9 had 0.5mg entecavir, 4 had 30mg clevudine, and 3 had 10mg adefovir. At the last follow up, these patients were on 0.5-1.0mg entecavir (n=23), 600mg telbivudine (n=9), 10mg adefovir (n=4), 300mg tenofovir (n=2), or combination therapy of lamivudine plus adefovir/tenofovir (n=2). Histology of the third biopsy showed complete resolution of interface hepatitis in 60 %of patients with the remainder

showing mild-to-moderate http://www.selleckchem.com/products/3-methyladenine.html activity. Persistent immunoreactivity for HBsAg was found in 80%, the mean number of hepatocytes positive for HBsAg being 10.4 %(range 1-80%). All but 1 (2.5%) was immunoreactive for HBcAg. At baseline, the median serum HBV DNA, HBsAg, ihHBV-DNA and cccDNA levels were 6.84 logIU/ mL, 3.38 logIU/mL, 286 copies/cell, and 7.3 copies/cell, respectively. At the time

of the last biopsies, 36 (90%) patients had undetectable serum HBV DNA (<20 IU/mL), all but one patient still had detectable HBsAg (median: 2.74 logIU/mL), all had detectable ihHBV-DNA (median: 0.4 copies/cell), but 18 (45%) patients had undetectable cccDNA. There was a trend of reduction of HBsAg, ihHBV-DNA and cccDNA levels from baseline to 1 year to last follow-up (all p<0.0001). The median Venetoclax log drop of HBsAg at last biopsy was 0.55 logIU/mL. The median percentage reductions of HBsAg, ihHBV-DNA and cccDNA at last biopsies were 71.46%, 99.85 %and 99.89%, respectively. Conclusions: Long-term NA treatment significantly reduced cccDNA and ihDNA. 45 %of patients had undetect-able

cccDNA, although small amount of ihHBV-DNA were still detectable in all patients. Integrated HBV DNA may be a possible source of detectable ihHBV-DNA and HBsAg. Continuous long-term NA therapy can reduce cccDNA to undetectable levels, suggesting a possible end-point of treatment. Disclosures: Ching-Lung Lai – Advisory Committees or Review Panels: Bristol-Myers Squibb, Gilead Sciences Inc; Consulting: Bristol-Myers Squibb, Gilead Sciences, Inc; Speaking and Teaching: Bristol-Myers Squibb, Gilead Sciences, Inc Wai-Kay Seto – Advisory Committees or Review Panels: click here Gilead Science; Speaking and Teaching: Gilead Science, Bristol-Myers Squibb Man-Fung Yuen – Advisory Committees or Review Panels: GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer; Grant/Research Support: Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmith-Kline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science The following people have nothing to disclose: Danny Wong, Philip Ip, Malgor-zata Kopaniszen, James Fung, Fung-Yu Huang, Brian P. Lee, Giuseppe Cullaro, Chi Hang Wu, Charles Cheng, Chi Hang J.

5 years (range: 6.0 – 11.9 years). At baseline, 13 patients had 100mg lamivudine, 11 had 600mg telbivudine, 9 had 0.5mg entecavir, 4 had 30mg clevudine, and 3 had 10mg adefovir. At the last follow up, these patients were on 0.5-1.0mg entecavir (n=23), 600mg telbivudine (n=9), 10mg adefovir (n=4), 300mg tenofovir (n=2), or combination therapy of lamivudine plus adefovir/tenofovir (n=2). Histology of the third biopsy showed complete resolution of interface hepatitis in 60 %of patients with the remainder

showing mild-to-moderate Rapamycin in vitro activity. Persistent immunoreactivity for HBsAg was found in 80%, the mean number of hepatocytes positive for HBsAg being 10.4 %(range 1-80%). All but 1 (2.5%) was immunoreactive for HBcAg. At baseline, the median serum HBV DNA, HBsAg, ihHBV-DNA and cccDNA levels were 6.84 logIU/ mL, 3.38 logIU/mL, 286 copies/cell, and 7.3 copies/cell, respectively. At the time

of the last biopsies, 36 (90%) patients had undetectable serum HBV DNA (<20 IU/mL), all but one patient still had detectable HBsAg (median: 2.74 logIU/mL), all had detectable ihHBV-DNA (median: 0.4 copies/cell), but 18 (45%) patients had undetectable cccDNA. There was a trend of reduction of HBsAg, ihHBV-DNA and cccDNA levels from baseline to 1 year to last follow-up (all p<0.0001). The median INCB024360 cost log drop of HBsAg at last biopsy was 0.55 logIU/mL. The median percentage reductions of HBsAg, ihHBV-DNA and cccDNA at last biopsies were 71.46%, 99.85 %and 99.89%, respectively. Conclusions: Long-term NA treatment significantly reduced cccDNA and ihDNA. 45 %of patients had undetect-able

cccDNA, although small amount of ihHBV-DNA were still detectable in all patients. Integrated HBV DNA may be a possible source of detectable ihHBV-DNA and HBsAg. Continuous long-term NA therapy can reduce cccDNA to undetectable levels, suggesting a possible end-point of treatment. Disclosures: Ching-Lung Lai – Advisory Committees or Review Panels: Bristol-Myers Squibb, Gilead Sciences Inc; Consulting: Bristol-Myers Squibb, Gilead Sciences, Inc; Speaking and Teaching: Bristol-Myers Squibb, Gilead Sciences, Inc Wai-Kay Seto – Advisory Committees or Review Panels: selleck kinase inhibitor Gilead Science; Speaking and Teaching: Gilead Science, Bristol-Myers Squibb Man-Fung Yuen – Advisory Committees or Review Panels: GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer; Grant/Research Support: Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmith-Kline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science The following people have nothing to disclose: Danny Wong, Philip Ip, Malgor-zata Kopaniszen, James Fung, Fung-Yu Huang, Brian P. Lee, Giuseppe Cullaro, Chi Hang Wu, Charles Cheng, Chi Hang J.

We speculate that triple therapy including JAK inhibitor telaprevir at the reduced dose of 1500 mg/day could maintain high levels of adherence to PEG IFN and RBV, and consequently

achieve high SVR rates. In this study, we investigated the independent predictors for SVR in the multivariate analysis (Table 3). As reported in previous studies, IL28B genotype remained the strongest predictor of SVR.[30, 31] The next strongest predictive factor was sex: women had significantly lower SVR rates than did men (Fig. 3). However, when we investigated the SVR rates of the telaprevir 2250 mg/day group and 1500 mg/day group, we found that there were significant differences in SVR rates between men and women in the telaprevir 2250 mg/day group but no differences in the telaprevir 1500 mg/day group. In the previous study, we reported that female sex was one of the factors influencing decreases in hemoglobin levels during triple therapy administrated 2250 mg/day of initial telaprevir dose.[20] In the present study, the discontinuation rates of telaprevir due to anemia were significantly higher in women in the telaprevir 2250 mg/day group as compared

with men (36.7% vs 3.3%, P = 0.002, data not shown), but there were no differences in the discontinuation rates of telaprevir due to anemia VX-809 manufacturer between men and women in the telaprevir 1500 mg/day group (0% vs 10%, P = 0.237, data not shown). Therefore, we speculate that there were significant differences in SVR rates between men click here and women because of high telaprevir discontinuation rates owing to anemia in women. In conclusion, after the completion of 24 weeks of therapy, triple therapy including telaprevir at a reduced dose of 1500 mg/day

was as effective as triple therapy including telaprevir 2250 mg/day at suppressing HCV RNA to undetectable levels and achieving SVR. Of note, we found that telaprevir 1500 mg/day was associated with lower levels of anemia and discontinuation of telaprevir owing to anemia, and higher PEG IFN and RBV adherence during triple therapy. These results suggest that the telaprevir 1500 mg/day regimen is an effective and safe alternative for the treatment of elderly and female Japanese patients. This study is a retrospective study. Prospective randomized controlled studies with longer follow-up periods are required to fully assess the efficacy and safety of an initial telaprevir dose of 1500 mg/day. THIS STUDY WAS supported in part by a Grant-in-Aid from the Ministry of Health, Labor and Welfare, Japan. “
“Tumor heterogeneity is a major obstacle for developing effective anticancer treatments. Recent studies have pointed to large stochastic genetic heterogeneity within cancer lesions, where no pattern seems to exist that would enable a more structured targeted therapy approach.

We speculate that triple therapy including selleck inhibitor telaprevir at the reduced dose of 1500 mg/day could maintain high levels of adherence to PEG IFN and RBV, and consequently

achieve high SVR rates. In this study, we investigated the independent predictors for SVR in the multivariate analysis (Table 3). As reported in previous studies, IL28B genotype remained the strongest predictor of SVR.[30, 31] The next strongest predictive factor was sex: women had significantly lower SVR rates than did men (Fig. 3). However, when we investigated the SVR rates of the telaprevir 2250 mg/day group and 1500 mg/day group, we found that there were significant differences in SVR rates between men and women in the telaprevir 2250 mg/day group but no differences in the telaprevir 1500 mg/day group. In the previous study, we reported that female sex was one of the factors influencing decreases in hemoglobin levels during triple therapy administrated 2250 mg/day of initial telaprevir dose.[20] In the present study, the discontinuation rates of telaprevir due to anemia were significantly higher in women in the telaprevir 2250 mg/day group as compared

with men (36.7% vs 3.3%, P = 0.002, data not shown), but there were no differences in the discontinuation rates of telaprevir due to anemia Fulvestrant solubility dmso between men and women in the telaprevir 1500 mg/day group (0% vs 10%, P = 0.237, data not shown). Therefore, we speculate that there were significant differences in SVR rates between men selleck and women because of high telaprevir discontinuation rates owing to anemia in women. In conclusion, after the completion of 24 weeks of therapy, triple therapy including telaprevir at a reduced dose of 1500 mg/day

was as effective as triple therapy including telaprevir 2250 mg/day at suppressing HCV RNA to undetectable levels and achieving SVR. Of note, we found that telaprevir 1500 mg/day was associated with lower levels of anemia and discontinuation of telaprevir owing to anemia, and higher PEG IFN and RBV adherence during triple therapy. These results suggest that the telaprevir 1500 mg/day regimen is an effective and safe alternative for the treatment of elderly and female Japanese patients. This study is a retrospective study. Prospective randomized controlled studies with longer follow-up periods are required to fully assess the efficacy and safety of an initial telaprevir dose of 1500 mg/day. THIS STUDY WAS supported in part by a Grant-in-Aid from the Ministry of Health, Labor and Welfare, Japan. “
“Tumor heterogeneity is a major obstacle for developing effective anticancer treatments. Recent studies have pointed to large stochastic genetic heterogeneity within cancer lesions, where no pattern seems to exist that would enable a more structured targeted therapy approach.

We speculate that triple therapy including GS-1101 solubility dmso telaprevir at the reduced dose of 1500 mg/day could maintain high levels of adherence to PEG IFN and RBV, and consequently

achieve high SVR rates. In this study, we investigated the independent predictors for SVR in the multivariate analysis (Table 3). As reported in previous studies, IL28B genotype remained the strongest predictor of SVR.[30, 31] The next strongest predictive factor was sex: women had significantly lower SVR rates than did men (Fig. 3). However, when we investigated the SVR rates of the telaprevir 2250 mg/day group and 1500 mg/day group, we found that there were significant differences in SVR rates between men and women in the telaprevir 2250 mg/day group but no differences in the telaprevir 1500 mg/day group. In the previous study, we reported that female sex was one of the factors influencing decreases in hemoglobin levels during triple therapy administrated 2250 mg/day of initial telaprevir dose.[20] In the present study, the discontinuation rates of telaprevir due to anemia were significantly higher in women in the telaprevir 2250 mg/day group as compared

with men (36.7% vs 3.3%, P = 0.002, data not shown), but there were no differences in the discontinuation rates of telaprevir due to anemia Protease Inhibitor Library between men and women in the telaprevir 1500 mg/day group (0% vs 10%, P = 0.237, data not shown). Therefore, we speculate that there were significant differences in SVR rates between men selleck chemicals llc and women because of high telaprevir discontinuation rates owing to anemia in women. In conclusion, after the completion of 24 weeks of therapy, triple therapy including telaprevir at a reduced dose of 1500 mg/day

was as effective as triple therapy including telaprevir 2250 mg/day at suppressing HCV RNA to undetectable levels and achieving SVR. Of note, we found that telaprevir 1500 mg/day was associated with lower levels of anemia and discontinuation of telaprevir owing to anemia, and higher PEG IFN and RBV adherence during triple therapy. These results suggest that the telaprevir 1500 mg/day regimen is an effective and safe alternative for the treatment of elderly and female Japanese patients. This study is a retrospective study. Prospective randomized controlled studies with longer follow-up periods are required to fully assess the efficacy and safety of an initial telaprevir dose of 1500 mg/day. THIS STUDY WAS supported in part by a Grant-in-Aid from the Ministry of Health, Labor and Welfare, Japan. “
“Tumor heterogeneity is a major obstacle for developing effective anticancer treatments. Recent studies have pointed to large stochastic genetic heterogeneity within cancer lesions, where no pattern seems to exist that would enable a more structured targeted therapy approach.

[28-30] Park et al.’s group reported that steatohepatitis, a common clinical condition, is a significant risk factor for HCC in a mouse model. In this case, the carcinogenic effect was mediated by interleukin (IL)−6 and TNF-α.[29] Furthermore, TNF-α was previously shown to be a significant contributor to inflammation Tyrosine Kinase Inhibitor Library screening in Mdr2-KO mice.[15] Considering that both TNF-α and IL-6 are secreted by macrophages, it is likely that these findings are very relevant to our results. The involvement of the CCR5 ligand, RANTES, in cancer has been studied mainly in breast cancer. In this disease, the majority of investigations claim a tumor-promoting role for RANTES.[31] RANTES levels were highly correlated with

advanced and progressed disease in breast cancer, and suggested

that the chemokine is directly involved in disease course. This hypothesis was proven correct in several studies that have manipulated the activities or expression of RANTES in animal model systems of breast cancer in mice. Different approaches—including the use of small iinterfering RNA to RANTES, the CCR5 antagonist, met-RANTES, and maraviroc, expression of the Δ32 CCR5, and overexpression of RANTES—have demonstrated that RANTES promotes tumor growth and disease progression.[32-35] Our results not only bolster the evidence that macrophages are indeed critical in inflammation-induced tumorigenesis, Selleck ABT 263 but also suggest that CCR5/RANTES axis is pivotal in their recruitment to the liver. It is conceivable that CCR5 is involved in several pathways of tumor development, including in both the inflammatory response that

induces oncogenic stress and the recruitment of cells that facilitates tumor progression and selleck products maintenance. Consequently, antagonists for CCR5 and CCR1, currently in clinical development, may prove useful in the prevention and treatment of liver inflammation, fibrosis, and HCC. Additional Supporting Information may be found in the online version of this article. “
“Rectal bleeding is a common complaint among adults presenting to doctors and emergency rooms. While the severity of bleeding can range from occult to massive, the patient is always worried. The clinical approach to the patient commences with an assessment of severity and type: occult, external, small volume, melena or maroon stool, large volume, or massive. Each type has a characteristic differential diagnosis. While colonoscopy is the mainstay of diagnosis, other testing that might be appropriate includes upper endoscopy, wireless capsule enteroscopy, push enteroscopy, balloon enteroscopy, red blood cell scintigraphy, and angiography, among others. “
“We read with interest the article by Sebastiani and colleagues regarding the use of SAFE biopsy in patients with chronic hepatitis C infection.1 Despite being a large and well-conducted study, there remain problems with the algorithm that may limit its use.

[28-30] Park et al.’s group reported that steatohepatitis, a common clinical condition, is a significant risk factor for HCC in a mouse model. In this case, the carcinogenic effect was mediated by interleukin (IL)−6 and TNF-α.[29] Furthermore, TNF-α was previously shown to be a significant contributor to inflammation this website in Mdr2-KO mice.[15] Considering that both TNF-α and IL-6 are secreted by macrophages, it is likely that these findings are very relevant to our results. The involvement of the CCR5 ligand, RANTES, in cancer has been studied mainly in breast cancer. In this disease, the majority of investigations claim a tumor-promoting role for RANTES.[31] RANTES levels were highly correlated with

advanced and progressed disease in breast cancer, and suggested

that the chemokine is directly involved in disease course. This hypothesis was proven correct in several studies that have manipulated the activities or expression of RANTES in animal model systems of breast cancer in mice. Different approaches—including the use of small iinterfering RNA to RANTES, the CCR5 antagonist, met-RANTES, and maraviroc, expression of the Δ32 CCR5, and overexpression of RANTES—have demonstrated that RANTES promotes tumor growth and disease progression.[32-35] Our results not only bolster the evidence that macrophages are indeed critical in inflammation-induced tumorigenesis, IWR-1 datasheet but also suggest that CCR5/RANTES axis is pivotal in their recruitment to the liver. It is conceivable that CCR5 is involved in several pathways of tumor development, including in both the inflammatory response that

induces oncogenic stress and the recruitment of cells that facilitates tumor progression and see more maintenance. Consequently, antagonists for CCR5 and CCR1, currently in clinical development, may prove useful in the prevention and treatment of liver inflammation, fibrosis, and HCC. Additional Supporting Information may be found in the online version of this article. “
“Rectal bleeding is a common complaint among adults presenting to doctors and emergency rooms. While the severity of bleeding can range from occult to massive, the patient is always worried. The clinical approach to the patient commences with an assessment of severity and type: occult, external, small volume, melena or maroon stool, large volume, or massive. Each type has a characteristic differential diagnosis. While colonoscopy is the mainstay of diagnosis, other testing that might be appropriate includes upper endoscopy, wireless capsule enteroscopy, push enteroscopy, balloon enteroscopy, red blood cell scintigraphy, and angiography, among others. “
“We read with interest the article by Sebastiani and colleagues regarding the use of SAFE biopsy in patients with chronic hepatitis C infection.1 Despite being a large and well-conducted study, there remain problems with the algorithm that may limit its use.

22 among rs8099917 GT/GG carriers versus 0.37 among TT carriers in the French cohort, P = 0.16, 0.52 versus 0.69 in the SCCS, P = 0.006; Fig. S1C). Similarly, the proportion of rapid fibrosis progressors differed according to IL28B rs8099917 in both cohorts, although significance was not

reached in the French cohort (proportion of rapid progressors 0.51 among GT/GG carriers versus 0.42 among TT carriers in the French cohort, P = 0.08; 0.56 versus 0.49 in the SCCS, P = 0.003; Fig. S1D). However, the effect differences in patients infected by HCV genotype 1 (Fig. S1E) versus genotype 3 (Fig. S1F) were less striking than those observed for the fibrosis stage. Similar but less significant results were found for rs12979860 (Fig. S2). Elevated ALT levels tended to be less frequent among patients BMS-777607 carrying the minor alleles of rs8099917 and rs12979860, but none of these differences were significant (Table S2), even after stratification by viral genotypes (Fig. 1B). Finally, no association was detected between IL28B SNPs and the development of HCC (Fig. S1).

Among patients with assessable response to treatment, a fibrosis stage ≥F2 was associated with a reduced sustained virologic response (SVR) (OR = 0.553, 95% CI 0.351-0.872, P = 0.01), but necroinflammatory activity (P = 0.7) and FPR (P = 0.7) were not. Using well-characterized chronic hepatitis C patients from two large cohorts, we showed that IL28B polymorphisms linked to a poor virological response to therapy are protective against liver necroinflammation and fibrosis progression, especially Selleckchem Sirolimus check details in patients with HCV genotypes other than 1. Previous observations on the role of IL28B polymorphisms with regard to necroinflammatory activity, fibrosis stage, transaminases, or gamma-glutamyl transpeptidase levels were reported from Japan,29 and, in abstract form, from the IDEAL trial performed in the U.S.30, 31 However, these studies were largely29 or exclusively30, 31 limited to patients infected with HCV genotype 1. A more recent work on a limited series of patients failed to show any association between IL28B genotype and

FPR.32 In the present study we analyzed the association of IL28B polymorphisms with necroinflammatory activity, ALT, fibrosis stage, and FPR in two large, well-pedigreed series of patients infected with the four most frequent HCV genotypes. Our data show a clear association between the poor treatment-response associated alleles of IL28B and low fibrosis stages as well as slow FPR in patients infected with HCV non-1 genotypes, but not in those infected with genotype 1, in agreement with the IDEAL and the Milan studies.32 A weak association between the poor treatment response allele of rs8099917 and low fibrosis stages was observed in the Japanese study, but the discrepancy may be explained by the contribution of genotype 2-infected patients, who were analyzed together with those infected with genotype 1.

22 among rs8099917 GT/GG carriers versus 0.37 among TT carriers in the French cohort, P = 0.16, 0.52 versus 0.69 in the SCCS, P = 0.006; Fig. S1C). Similarly, the proportion of rapid fibrosis progressors differed according to IL28B rs8099917 in both cohorts, although significance was not

reached in the French cohort (proportion of rapid progressors 0.51 among GT/GG carriers versus 0.42 among TT carriers in the French cohort, P = 0.08; 0.56 versus 0.49 in the SCCS, P = 0.003; Fig. S1D). However, the effect differences in patients infected by HCV genotype 1 (Fig. S1E) versus genotype 3 (Fig. S1F) were less striking than those observed for the fibrosis stage. Similar but less significant results were found for rs12979860 (Fig. S2). Elevated ALT levels tended to be less frequent among patients selleck inhibitor carrying the minor alleles of rs8099917 and rs12979860, but none of these differences were significant (Table S2), even after stratification by viral genotypes (Fig. 1B). Finally, no association was detected between IL28B SNPs and the development of HCC (Fig. S1).

Among patients with assessable response to treatment, a fibrosis stage ≥F2 was associated with a reduced sustained virologic response (SVR) (OR = 0.553, 95% CI 0.351-0.872, P = 0.01), but necroinflammatory activity (P = 0.7) and FPR (P = 0.7) were not. Using well-characterized chronic hepatitis C patients from two large cohorts, we showed that IL28B polymorphisms linked to a poor virological response to therapy are protective against liver necroinflammation and fibrosis progression, especially ABT 263 find more in patients with HCV genotypes other than 1. Previous observations on the role of IL28B polymorphisms with regard to necroinflammatory activity, fibrosis stage, transaminases, or gamma-glutamyl transpeptidase levels were reported from Japan,29 and, in abstract form, from the IDEAL trial performed in the U.S.30, 31 However, these studies were largely29 or exclusively30, 31 limited to patients infected with HCV genotype 1. A more recent work on a limited series of patients failed to show any association between IL28B genotype and

FPR.32 In the present study we analyzed the association of IL28B polymorphisms with necroinflammatory activity, ALT, fibrosis stage, and FPR in two large, well-pedigreed series of patients infected with the four most frequent HCV genotypes. Our data show a clear association between the poor treatment-response associated alleles of IL28B and low fibrosis stages as well as slow FPR in patients infected with HCV non-1 genotypes, but not in those infected with genotype 1, in agreement with the IDEAL and the Milan studies.32 A weak association between the poor treatment response allele of rs8099917 and low fibrosis stages was observed in the Japanese study, but the discrepancy may be explained by the contribution of genotype 2-infected patients, who were analyzed together with those infected with genotype 1.

Moreover, quantification of intracellular Akt inhibitor HBV DNA at an MOI of 50 demonstrated that the magnitude of HBV replication in PMHs was 106 HBV DNA copies per well; it reached a peak around day 3 PT and

lasted more than 9 days (Fig. 1E). In comparison, the transduction of HepG2 cells was more efficient because intracellular HBV DNA quantification was 2 log higher than that in PMHs (Fig. 1E). Moreover, by using a recombinant baculovirus for green fluorescent protein, we found that the transduction efficiency at an MOI of 50 was clearly higher in HepG2 cells versus PMHs, with more than 70% and 50% of the cells transduced, respectively (data not shown). When we looked at extracellular HBV DNA after PMH transduction, we observed a peak of secretion on day 6 PT with 8 × 105 copies per well (Fig. 1F). Finally, the results also showed that the amount of the intracellular baculovirus genome was maximum on day 1 PT, and it decreased thereafter but was still detectable by Southern blot analysis on day 9 PT (Fig. 1C). The differences in the kinetics of accumulation and clearance of both baculoviral and HBV nucleic acids suggest that

the half-life of encapsidated HBV DNA is longer than that of the baculoviral genome, as previously described.12 It has previously been shown that HepG2 cells transduced with HBV baculoviruses produce infectious HBV particles.12, selleck 18 The supernatant from HBV baculovirus–transduced PMHs was first characterized by CsCl gradient analyses. Most HBV DNA was found in fractions with a density range of 1.24 and 1.20 g/cm3 (fractions 9-12; Fig. 2A). This indicates that most HBV DNA was released within Dane particles, which have been shown to peak at 1.21 g/cm3 in isopycnic density analysis.18 Electron microscopy analyses confirmed the presence of Dane particles as well as spheres and filamentous particles (Fig. 2B). Altogether, these data showed that HBV-transduced

PMHs secreted newly produced HBV particles in 3 typical forms.19 Concentrated supernatant from HBV baculovirus–transduced PMHs was then used to inoculate freshly prepared PMHs or HepaRG cells,20 but none of these cells showed convincing signals of HBV infection within 15 days after inoculation (data not shown). The last step was the evaluation of the potentiality of our system learn more for antiviral therapy testing. First, the antiviral activity of lamivudine was tested. Nucleos(t)ide analogues such as lamivudine are able to specifically inhibit HBV viral polymerase activity and thus prevent the secretion of mature HBV particles, whereas other steps of the viral life cycle, such as entry or antigen secretion, are not targeted by such a treatment.21 As expected, HBV DNA secretion (Fig. 3A), but not HBsAg (Fig. 3B), was inhibited by lamivudine treatment in a dose-dependent manner in PMHs transduced with Bac-HBV-1.