Cause Of Chronic Hepatitis Research Articles

Non-Coding RNAs and Hepatitis C Virus-Induced Hepatocellular Carcinoma.

Hepatitis C virus (HCV) infection is a worldwide health problem and is one of the main causes of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). Despite recent improvements, effective treatments for HCC are still missing and new tools for early detection are needed. Non-coding RNAs (ncRNAs) have emerged as important regulators of gene expression and key players in human carcinogenesis, including HCC. Aberrant expression of ncRNAs is associated with HCC metastasis, invasion, dissemination, and recurrence. This review will focus on the recent advances in ncRNA expression profiles, their dysregulation in HCV-related HCC, and the clinical perspective of ncRNA signatures for the early detection of HCC.

Efficacy of tenofovir in preventing perinatal transmission of HBV infection in pregnant women with high viral loads

Mother-to-child transmission is the major cause of chronic hepatitis B virus (HBV) infection. This double-blind trial tested the effect of tenofovir disoproxil fumarate (TDF) in preventing vertical transmission. Pregnant women who were HBsAg/HBeAg-positive with a HBV DNA titer ≥ 2×106 IU/mL were randomly assigned to the control (n = 60) and TDF-treated (n = 60) groups. TDF treatment (oral dose 300 mg/day) was initiated at 24 weeks of gestation and continued to 4 weeks after delivery. The subjects were followed up to 28 weeks postpartum. The effects of TDF on vertical transmission, outcomes of the mothers and infants and virological changes were monitored. TDF dynamically reduced the serum HBV DNA level of the mothers, particularly during the first 4 weeks of treatment. The lower viral loads were maintained in the pregnancies until delivery. Approximately 90% and 33.9% of the TDF-treated mothers had viral loads ≤2000 IU/mL after delivery and at 28 weeks postpartum, respectively. No cervical transmission or adverse effects were observed in the TDF-treated individuals, whereas 13.5% of the infants were infected with HBV in the control group. We conclude that TDF treatment initiated at 24 weeks of gestation in high-viremia, HBsAg/HBeAg-positive mothers efficiently prevents mother-to-child HBV transmission without adverse events in mothers and infants.

Impact of DAA-Based Therapy on Alfa-Fetoprotein Level in Chronic Hepatitis C Patients

Introduction: Backgrounds: Chronic hepatitis C virus (HCV) infection is a leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma worldwide. High AFP level has been identified as a risk factor for the development of HCC in chronic hepatitis C patients. The aim of this study is to investigate the impact of DAA-based triple therapy on serum AFP levels of patients with chronic hepatitis C. Methods: We reviewed records of 132 consecutive patients with chronic hepatitis C who received DAAbased therapy over past 16 months. Inclusion criteria: patients who completed treatment, had baseline AFP within 3 months of treatment, and AFP within 6 months post-treatment. 54 patients were excluded (52: no AFP level, and 2 patient were lost to follow). Results: 78 patients completed the treatment: age: 24-78 year, 45 males, 33 females. Of these patients: 46 genotype 1, 22 genotype 2, 10 genotypes 3 and 4. All had viral loads, and fibrosis score. 76 patients achieved SVR at 3 months and 6 months pots-treatment. 2 patients relapsed. Conclusion: Discussion: AFP is commonly used for surveillance for HCC. In this review, there was universal and lasting ( beyond 6 months posttreatment) reduction in AFP below 50% of baseline level in all patients s who received treatment even in those who relapsed, and across all genotypes, most pronounced in patients with markedly high pretreatment level and those with documented HCC. This phenomenon supports the evidence that DAA therapy may prevent HCC in hepatitis C patients. Conclusion: DAA therapy of hepatitis C-infected patient effectively reduce the level of AFP and so potentially may prevent HCC in this group of patients2851_A Figure 1. AFP: Pre- and posttreatment

Adaptive mutation F772S-enhanced p7-NS4A cooperation facilitates the assembly and release of hepatitis C virus and is associated with lipid droplet enlargement

Hepatitis C virus (HCV) infection is a major cause of chronic hepatitis and liver cancer worldwide. Adaptive mutations play important roles in the development of the HCV replicon and its infectious clones. We and others have previously identified the p7 mutation F772S and the co-presence of NS4A mutations in infectious HCV full-length clones and chimeric recombinants. However, the underlying mechanism of F772S function remains incompletely understood. Here, we investigated the functional role of F772S using an efficient JFH1-based reporter virus with Core-NS2 from genotype 2a strain J6, and we designated J6-p7/JFH1-4A according to the strain origin of the p7 and NS4A sequences. We found that replacing JFH1-4A with J6-4A (wild-type or mutated NS4A) or genotype 2b J8-4A severely attenuated the viability of J6-p7/JFH1-4A. However, passage-recovered viruses that contained J6-p7 all acquired F772S. Introduction of F772S efficiently rescued the viral spread and infectivity titers of J6-p7/J6-4A, which reached the levels of the original J6-p7/JFH1-4A and led to a concomitant increase in RNA replication, assembly and release of viruses with J6-specific p7 and NS4A. These data suggest that an isolate-specific cooperation existed between p7 and NS4A. NS4A exchange- or substitution-mediated viral attenuation was attributed to the RNA sequence, and no p7-NS4A protein interaction was detected. Moreover, we found that F772S-enhanced p7-NS4A cooperation was associated with the enlargement of intracellular lipid droplets. This study therefore provides new insights into the mechanisms of adaptive mutations and facilitates studies on the HCV life cycle and virus–host interaction.

Hepatitis C Virus Infection in a Tertiary Care Hospital in Mumbai, India: Identification of a Mixed and Novel Genotype

Purpose: Hepatitis C virus (HCV) is a leading cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC). HCV being a ribonucleic acid virus has considerable sequence variability. Assessment of viral load and genotype is necessary for designing treatment strategies and monitoring for viral resistance among HCV-infected cases. HCC is the most common form of liver cancer, often occurring in people with chronic hepatitis B or C. We undertook this study to observe genotype distribution of the virus in HCV patients from Mumbai. Materials and Methods: Between January 2017 and December 2017, the study was conducted on 120 chronic hepatitis outpatients from a tertiary care hospital, Mumbai, after obtaining ethics approval. All these diagnosed cases of HCV were subjected to molecular diagnosis in a research institute, Mumbai, by real-time polymerase chain reaction-based techniques. Results: Males were more preponderant than females with HCV infection, and the highest number of HCV-infected cases was observed in the age group of 41–50 years. Genotype 3 (n = 70; 58.3%) accounted for the highest number of cases followed by genotypes 1b (n = 29; 24.2%) and then 1a (n = 14; 11.7%). Mixed genotypes 1b + 3 and individual genotype 4 were found in two cases each (1.7%). A total of three samples (2.5%) were found with untypeable genotype. Conclusion: The major HCV genotype observed was 3 which is difficult to treat with direct-acting antivirals, owing to the more rapid progression of liver disease, increased rates of steatosis (non-alcoholic fatty liver disease), a higher risk for cancer (HCC). We believe this study is the first one to address the prevalence of mixed genotypes and untypeable genotype from India.

Diagnostic value of combined serum biomarkers for the evaluation of liver fibrosis in chronic hepatitis C infection: A multicenter, noninterventional, observational study

The hepatitis C virus (HCV) infection is important cause of chronic hepatitis. Liver biopsy is considered the gold standard for assessment of fibrosis but this procedure is an invasive procedure. We aimed to evaluate the diagnostic efficiency of non-invasive serum biomarkers, separately and in combinations, on liver fibrosis in treatment-naive chronic hepatitis C (CHC) patients. Two hundred and sixteen treatment-naive CHC patients were enrolled from 32 locations across Turkey in this open-labelled, non-interventional prospective observational study. FibroTest®, aspartate aminotransferase-to-platelet ratio index(APRI), aspartate aminotransferase and alanine aminotransferase ratio (AAR), fibrosis index based on four factors (FIB-4), Age-platelet(AP) index and Forns index were measured and compared with Metavir scores got from liver biopsies. Data from 182 patients with baseline liver biopsy were suitable for analysis. One hundred and twenty patients (65.9%) had F0-F1 fibrosis and 62 patients (34.1%) had F2-F4 fibrosis. APRI 0.732 area under the curve(AUC) indicated advanced fibrosis with 69% sensitivity and 77% specificity. FIB-4 0.732 AUC and FibroTest 0.715 AUC indicated advanced fibrosis with 69% and 78.4% sensitivity, and 75% and 71.4% specificity, respectively. The combined use of tests also led to an increase in AUC and specificity. Combinations of FibroTest with APRI and/or FIB-4, and FIB-4 with APRI were optimal for the evaluation of liver fibrosis. Fibrotest, FIB-4, APRI, AP index and Forns index exhibit good diagnostic performance for determining liver fibrosis in CHC patients, and the use of at least two tests together will increase their diagnostic value still further.

Structure-Guided Combinatorial Engineering Facilitates Affinity and Specificity Optimization of Anti-CD81 Antibodies

Hepatitis C viral infection is the major cause of chronic hepatitis that affects as many as 71 million people worldwide. Rather than target the rapidly shifting viruses and their numerous serotypes, four independent antibodies were made to target the host antigen CD81 and were shown to block hepatitis C viral entry. The single-chain variable fragment of each antibody was crystallized in complex with the CD81 large extracellular loop in order to guide affinity maturation of two distinct antibodies by phage display. Affinity maturation of antibodies using phage display has proven to be critical to therapeutic antibody development and typically involves modification of the paratope for increased affinity, improved specificity, enhanced stability or a combination of these traits. One antibody was engineered for increased affinity for human CD81 large extracellular loop that equated to increased efficacy, while the second antibody was engineered for cross-reactivity with cynomolgus CD81 to facilitate animal model testing. The use of structures to guide affinity maturation library design demonstrates the utility of combining structural analysis with phage display technologies.

DIAGNOSIS MOLEKUL DAN APLIKASI DALAM PENGOBATAN HEPATITIS B & C

In the whole world, up to now hepatitis B and C virus are still the main causes of chronic hepatitis, cirrhosis until hepatocellularcarcinoma. Most diagnosis is based on a serological examination such as the determination of antigen and antibody, for exampleHbsAg, HbeAg and anti HCV. Recently, for hepatitis C examination, HCV core antigen is used for the detection of HCV infection duringthe window period, chronic C hepatitis and for treatment monitoring. At present, serological assays are not sufficient to confirm thediagnosis of hepatitis B and C due to mutations of false negative HbsAg or HbeAg results. Occult hepatitis B can occur with a negativeresult of HbsAg, which cause difficulties in confirming the diagnosis and treatment as well. The success of treatment can be influencedof both hepatitis B and hepatitis C that have genotypes. By using molecular examination, such as determination of HBV-DNA and HCVRNA,it is expected that the problem of serological determination can be overcome. Molecular examination is not only useful for just thediagnosis confirmation, such as for active phase and replicative determination. This sequence is also very useful as a data base prior tothe treatment of hepatitis B and hepatitis C as well as for their following success result.

Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry.

Hepatitis C virus (HCV) is the leading cause of chronic hepatitis in humans. Several host molecules participate in HCV cell entry, but this process remains unclear. The complete unraveling of the HCV entry process is important to further understand viral pathogenesis and develop therapeutics. Human hepatitis A virus (HAV) cellular receptor 1 (HAVCR1), CD365, also known as TIM-1, functions as a phospholipid receptor involved in cell entry of several enveloped viruses. Here, we studied the role of HAVCR1 in HCV infection. HAVCR1 antibody inhibited entry in a dose-dependent manner. HAVCR1 soluble constructs neutralized HCV, which did not require the HAVCR1 mucinlike region and was abrogated by a mutation of N to A at position 94 (N94A) in the Ig variable (IgV) domain phospholipid-binding pocket, indicating a direct interaction of the HAVCR1 IgV domain with HCV virions. However, knockout of HAVCR1 in Huh7 cells reduced but did not prevent HCV growth. Interestingly, the mouse HAVCR1 ortholog, also a phospholipid receptor, did not enhance infection and a soluble form failed to neutralize HCV, although replacement of the mouse IgV domain with the human HAVCR1 IgV domain restored the enhancement of HCV infection. Mutations in the cytoplasmic tail revealed that direct HAVCR1 signaling is not required to enhance HCV infection. Our data show that the phospholipid-binding function and other determinant(s) in the IgV domain of human HAVCR1 enhance HCV infection. Although the exact mechanism is not known, it is possible that HAVCR1 facilitates entry by stabilizing or enhancing attachment, leading to direct interactions with specific receptors, such as CD81.IMPORTANCE Hepatitis C virus (HCV) enters cells through a multifaceted process. We identified the human hepatitis A virus cellular receptor 1 (HAVCR1), CD365, also known as TIM-1, as a facilitator of HCV entry. Antibody blocking and silencing or knockout of HAVCR1 in hepatoma cells reduced HCV entry. Our findings that the interaction of HAVCR1 with HCV early during infection enhances entry but is not required for infection support the hypothesis that HAVCR1 facilitates entry by stabilizing or enhancing virus binding to the cell surface membrane and allowing the correct virus-receptor positioning for interaction with the main HCV receptors. Furthermore, our data show that in addition to the phospholipid-binding function of HAVCR1, the enhancement of HCV infection involves other determinants in the IgV domain of HAVCR1. These findings expand the repertoire of molecules that HCV uses for cell entry, adding to the already complex mechanism of HCV infection and pathogenesis.

Hepatitis C: from discovery to cure

Background and ObjectivesWith a prevalence of approximately 71 million infected individuals globally, chronic hepatitis C virus (HCV) infection is one of the major causes of chronic hepatitis leading to hepatocellular carcinoma (HCC). This review aimed to summarize the general history of HCV and provide a perspective on future journey of HCV.Materials and MethodsWe searched for articles published in periodicals, monographs and edited books with the key words including ‘non‐A, non‐B hepatitis’, ‘HCV’, ‘interferon’ and ‘direct‐antiviral agents (DAAs)’. And the data emphasized HCV emergence, identification, treatment, epidemiology as well as the cellular and animal models leading to the eradication of HCV infections, were summarized.Results and ConclusionThe battle against hepatitis C is destined to be recorded in history as one of science's remarkable success. Although the revolutionary direct‐antiviral agents (DAAs) are able to cure more than 95% of HCV patients, access to diagnosis and therapy remains improved. More efforts should be made to promote HCV screening, treatment delivery and vaccine development.

The impact of treatment of hepatitis C with DAAs on the occurrence of HCC.

Hepatitis C virus infection is a major cause of chronic hepatitis resulting in cirrhosis and hepatocellular carcinoma (HCC). The recent introduction of direct acting antivirals (DAA), results in sustained virological response (SVR) rates of >90% in treated patients whatever the stage of liver fibrosis with an excellent safety profile. This major advancement has allowed treatment of a larger number of patients, some with more advanced liver dysfunction and a higher risk of HCC. An SVR is associated with a reduced risk of hepatic decompensation, the need for liver transplantation and both liver-related and overall mortality. This high rate of SVR has raised hopes that there would be a significant reduction in the incidence of HCC. However, the impact of DAA-based regimens on the occurrence of HCC in patients with cirrhosis, and in particular the recurrence of HCC following successful curative treatment is controversial. Published studies suggest that DAA does not increase the risk of de novo HCC following SVR. A more controversial topic is the effect of a DAA-based SVR on the recurrence of HCC following curative treatment of early HCC. Well-designed studies with robust comparisons are needed to determine the effect of DAA on the recurrence of HCC. At present, patients with HCV cirrhosis who have undergone resection or ablation for HCC should not be dissuaded from receiving DAA therapy to prevent the progression of liver disease. Monitoring for HCC with liver imaging and AFP should be performed twice a year indefinitely post-SVR in patients with HCV cirrhosis.

N-Myc Downstream-Regulated Gene 1 Restricts Hepatitis C Virus Propagation by Regulating Lipid Droplet Biogenesis and Viral Assembly.

Host cells harbor various intrinsic mechanisms to restrict viral infections as a first line of antiviral defense. Viruses have evolved various countermeasures against these antiviral mechanisms. Here we show that N-Myc downstream-regulated gene 1 (NDRG1) limits productive hepatitis C virus (HCV) infection by inhibiting viral assembly. Interestingly, HCV infection downregulates NDRG1 protein and mRNA expression. The loss of NDRG1 increases the size and number of lipid droplets, which are the sites of HCV assembly. HCV suppresses NDRG1 expression by upregulating MYC, which directly inhibits the transcription of NDRG1 The upregulation of MYC also leads to the reduced expression of the NDRG1-specific kinase serum/glucocorticoid-regulated kinase 1 (SGK1), resulting in a markedly diminished phosphorylation of NDRG1. The knockdown of MYC during HCV infection rescues NDRG1 expression and phosphorylation, suggesting that MYC regulates NDRG1 at both the transcriptional and posttranslational levels. Overall, our results suggest that NDRG1 restricts HCV assembly by limiting lipid droplet formation. HCV counteracts this intrinsic antiviral mechanism by downregulating NDRG1 via a MYC-dependent mechanism.IMPORTANCE Hepatitis C virus (HCV) is an enveloped single-stranded RNA virus that targets hepatocytes in the liver. HCV is a leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma, and estimates suggest a global prevalence of 2.35%. Up to 80% of acutely infected individuals will develop chronic infection, and as many as 5% eventually progress to liver cancer. An understanding of the mechanisms behind virus-host interactions and viral carcinogenesis is still lacking. The significance of our research is that it identifies a previously unknown relationship between HCV and a known tumor-associated gene. Furthermore, our data point to a new role for this gene in the liver and in lipid metabolism. Thus, HCV infection serves as a great biological model to advance our knowledge of liver functions and the development of liver cancer.

Sofosbuvir/simeprevir versus sofosbuvir/ribavirin regimen in Egyptian patients with hepatitis C virus

Background/aimHepatitis C virus (HCV) genotype 4 is the most frequent cause of chronic hepatitis in Egypt. The aim of this study was to compare the efficacy of sofosbuvir/simeprevir for 12 weeks versus sofosbuvir/ribavirin for 24 weeks in Egyptian patients with HCV-related Child A cirrhosis.MethodsN

Epidemiology of HBV in pregnant women, South West Nigeria

Hepatitis B virus (HBV) remains a leading cause of chronic hepatitis, maternal complications, and neonatal deaths in sub-Saharan Africa. Pregnant women serve as a major reservoir for the persistence and ongoing transmission of hepatitis B virus and HIV in a generalized heterosexual epidemic. The aim of this study is to assess the epidemiology of Hepatitis B infection among pregnant women in South West-Nigeria. This is a cross-sectional study of 353 pregnant women across 10 health facilities in the region. Results showed that of the 353 pregnant women tested, 37 were positive for the HBV antigen giving a prevalence estimate of 10.5% (95% CI: 7.5%–14.2%). We found significant negative association between odds of HBV infection and knowledge of HBV transmission through sex (OR: 0.30: 95%CI–0.11–0.82) and a positive association with blood transfusion in the past three months (OR: 9.5: 95% CI-1.58–57.14). Findings strongly suggest high endemicity of HBV and the possible implication of blood transfusion as a major route of ongoing HBV transmission among pregnant women in south-western Nigeria. We recommend further study of a prospective design to investigate the possible causal link between blood transfusion and the risk of HBV infection among pregnant women in Nigeria.

Comparison of Real Time PCR, Chemilluminicensce, ELISA and Rapid Immunochromatographic Tests for Detection of Hepatitis B Virus Infection

Background: Hepatitis B virus (HBV) is one of the main causes of chronic hepatitis in developing countries. The current study was done to evaluate the efficacy of the chemiluminescence, ELISA and immune chromatographic (ICT) test to Real Time-PCR (RT_PCR) for establishing the diagnosis of hepatitis B virus (HBV) in indoor and outdoor patients of Department of Medicine, Rajendra Institute of Medical Sciences. Methods and Material: This descriptive, cross-sectional study was carried out in 475 patients. The demographic data and patient’s serum samples of the subjects was collected. The samples were analysed by chemiluminescence, ELISA, ICT & real RT-PCR for HBV. Result: In the present study, of total 70 by chemiluminescence, 64 by ELISA, 61 by ICT and 63 by RT PCR were found positive for HBV. By both RT PCR and chemiluminescence 63 samples were positive. Total 2 patients were positive for HBV by RT PCR and negative by chemiluminescence while 7 patients were negative for RT-PCR and positive by chemiluminescence. Total 8 patients were positive for HBV by RTPCR and negative by ELISA while 5 patients were negative for RT- PCR and positive for ELISA. Total 11 patients were positive for HBV by RT PCR and negative by ICT while 3 patients were negative for RT- PCR and positive for ICT. On the basis of above findings chemiluminescence, ELISA and ICT specificity was 98.32%,98.79%,99.27%, sensitivity was 97.01%,89.04%,52% , positive predictive value was 90.27%, 92.85 %,95.58% and negative predictive value was 99.51%,98.08%,97.38% . Conclusion: The HBsAg negative patients should also be screened with HBc total antibody for suspecting the exposure which could be later confirmed by RT PCR .RT-PCR method is accredited as a specific and reliable method suitable for screening of HBV and should be recommended for establishing exact and final diagnosis of these patients.

Genotype 4 hepatitis E virus is a cause of chronic hepatitis in renal transplant recipients in Hong Kong

Genotype 4 hepatitis E virus is a cause of chronic hepatitis in renal transplant recipients in Hong Kong

Nghiên cứu sự lưu hành và kiểu gen của vi rút viêm gan B ở một số địa phương giai đoạn 2012 – 2013

The hepatitis B virus (HBV) is among the leading causes of chronic hepatitis, cirrhosis and hepatocellular car¬cinoma. Screening of 664 serum samples from Hoabinh, Quangtri, Kontum provinces and Ho Chi Minh city showed that the prevalence of HBsAg varied by geographic regions and gender. The highest one was in Hoabinh (18.27% in general, 21.95% in male, and 13.38% in female), and the lowest one was in Kontum (3.1% in general, 2.32% in male, and 4.32% in female). In Vietnam, the most common genotype is B, followed by genotypes C and G. HBV phylogenetic analysis of S sequences revealed that genotype B was highly prevalent (83.3%), and genotype C (8.3%) and G (8.3%) were also detected as a minor distribution. Further studies are recommended to determine the HBsAg prevalence and genotypes and main modes of hepatitis B virus transmission in Vietnam.

A model to predict the onset of non-alcoholic fatty liver disease within 2years in elderly adults.

Non-alcoholic fatty liver disease (NAFLD) is a common cause of chronic hepatitis, which leads to cirrhosis and hepatocellular carcinoma. However, it is difficult to identify subjects at high risk for NAFLD onset. This study aims to construct a model to predict the onset of NAFLD within 2years in elderly adults. This study included and followed 3378 initial NAFLD-free subjects aged 60years or over for 2years, which were randomly divided into a training set and a validation set. NAFLD was diagnosed on ultrasound. Clinical and laboratory data were recorded at baseline. A model was constructed in the training set to predict the onset of NAFLD and validated in the validation set. Body mass index, hemoglobin, fasting blood glucose, and triglycerides were identified as predictors for the onset of NAFLD. A risk score (R) was calculated by them. It classified the subjects into low-risk group (R≤-2.88), moderate-risk group (-2.88<R≤-1.26), and high-risk group (R>-1.26). In the training set, 4.68% of the participants in the low-risk group, 11.59% of the participants in the moderate-risk group, and 31.02% of the participants in the high-risk group developed NAFLD. In the validation set, 5.84% of the participants in the low-risk group, 10.57% of the participants in the moderate-risk group, and 29.44% of the participants in the high-risk group developed NAFLD. This study developed a model to predict the onset of NAFLD in elderly adults, which might provide indications for intervention to these subjects.

Association of human platelet antigens polymorphisms with susceptibility to hepatitis C virus infection in Chinese population.

Hepatitis C virus (HCV) is a major cause of chronic hepatitis. Previous studies have identified a number of single nucleotide polymorphisms that are associated with HCV infection. Human platelet antigens (HPAs) polymorphisms play an important role in several diseases. Here, we demonstrated the association of the HPA-2, HPA-3, HPA-5 and HPA-15 polymorphisms with susceptibility to HCV infection in Chinese population. Overall, 118 patients with HCV and 167 controls were genotyped for HPAs. There were no significant differences in the allele and genotype frequency distribution for the HPA-3, HPA-5 and HPA-15 systems between the patients with chronic HCV infection and the healthy controls (p>.05). However, the genotype frequency of HPA-2aa was significantly lower, while HPA-2ab/bb was significantly higher in patients than that in the controls (p=.006). The allele frequency of HPA-2a in patients was significantly lower than that in the control group (p=.005). In contrast, HPA-2b in patients was significantly higher than that in the control group (p=.005). We conclude that HPA-2 polymorphism is associated with susceptibility to HCV infection, and individuals carrying the HPA-2b allele may have a higher risk of HCV infection compared with individuals carrying HPA-2a.

Role of Cell-Penetrating Peptides in Intracellular Delivery of Peptide Nucleic Acids Targeting Hepadnaviral Replication.

Peptide nucleic acids (PNAs) are potentially attractive antisense agents against hepatitis B virus (HBV), although poor cellular uptake limits their therapeutic application. In the duck HBV (DHBV) model, we evaluated different cell-penetrating peptides (CPPs) for delivery to hepatocytes of a PNA-targeting hepadnaviral encapsidation signal (ε). This anti-ε PNA exhibited sequence-specific inhibition of DHBV RT in a cell-free system. Investigation of the best in vivo route of delivery of PNA conjugated to (D-Arg)8 (P1) showed that intraperitoneal injection to ducklings was ineffective, whereas intravenously (i.v.) injected fluorescein-P1-PNA reached the hepatocytes. Treatment of virus carriers with i.v.-administered P1-PNA resulted in a decrease in viral DNA compared to untreated controls. Surprisingly, a similar inhibition of viral replication was observed in vivo as well as in vitro in primary hepatocyte cultures for a control 2 nt mismatched PNA conjugated to P1. By contrast, the same PNA coupled to (D-Lys)4 (P2) inhibited DHBV replication in a sequence-specific manner. Interestingly, only P1, but not P2, displayed anti-DHBV activity in the absence of PNA cargo. Hence, we provide new evidence that CPP-PNA conjugates inhibit DHBV replication following low-dose administration. Importantly, our results demonstrate the key role of CPPs used as vehicles in antiviral specificity of CPP-PNA conjugates.