IRES complexity before IFN-alpha treatment and evolution of the viral load at the early stage of treatment in peripheral blood mononuclear cells from chronic hepatitis C patients.

In the proof, we asked to correct the name of one Author, and the adddess; this was acknowledged by the Editor, but was omitted in the final published version!International audienceAt the early stage of treatment, IFN alpha-2a induces inhibition of HCV replication. The viral load reflects mainly the degradation rate of the viruses. However, differences in the behavior of the viral population depend on changes, which occurred in the HCV-IRES genome. In this study, cloning and sequencing strategies permitted the generation of a large number of IRES sequences from the PBMCs of 18 patients (5 women, 13 men) with chronic hepatitis C. The HCV IRES appeared to be highly conserved structurally. However, some variability was found between the different isolates obtained: 467 substitutions with a median of 7 variants/patients. No relationship was observed between pre-treatment IRES complexity and the viral load at the beginning. However, on review of the evolution of viral load in the PBMCs during the first 3 days of IFN alpha-2a treatment, patients could be classified into two groups: Group 1, in which the viral population continued to replicate and Group 2, in which the viral load decreased significantly (P = 0.01727). Positioning of the mutations on the predicted IRES secondary structure showed that the distribution of the mutations and their apparition frequency were different between the two groups. At the early stage of treatment, IFN alpha-2a was efficient in reducing the viral replication in a significant number of patients; mechanisms of response might affect the virus directly. However, pre-treatment genomic variations observed in the 5'NCR of HCV were not a parameter of a later response to antiviral therapy in chronic hepatitis C patients. (244

Molecular epidemiology of hepatitis C virus genotypes in Bushehr province, Iran

Background and Objectives: Molecular epidemiology of hepatitis C virus (HCV) is very important for the treatment of hepatitis C infection. The aim of this study was to determine the distribution of HCV genotypes in Bushehr province (South West of Iran). Materials and Methods: A total of 100 patients who were detected as positive for HCV antibody (by using ELISA method and RIBA test) referred to Arya Virology Laboratory between 2007-2009 in order to molecular diagnosis and furthermore virus genotyping. After detection of HCV, RNA genotyping of virus was done by using genotype specific primers. Results: Genotype 1a was found in 49% of the patients and genotype 3a was found in 40% of the patients and 1b in 5% of patients, while the genotype of the virus could not be identified in 5% of the patients. Finally, in 1% of patients coinfection due to 1a-3a genotypes was identified. Conclusion: The dominant genotype of HCV in Bushehr province, Iran, was determined as 1a.with acute hepatitis C ultimately develop chronic infection1. Only a minority of cases of acute HCV recover completely, with spontaneous virus eradication. In most cases the acute infection progresses to chronicity. Chronic HCV infection is defined as an infection that persists for more than 6 months, with or without clinical manifestations of hepatic or extrahepatic disease. Chronic type of this infection can cause cirrhosis, liver failure, and liver cancer. HCV infection is a global health problem and it is estimated that 200 million people of the world population are infected5. The global spread of chronic HCV infection coincided with the widespread use of transfused blood and blood products and with the expansion of intravenous drug use but decreased prior to the wide implementation of anti-HCV screening6. There are at least six major genotypes designated by Arabic numerals and more than 50 subtypes of HCV identified by lower case letters. The different genotypes have different geographic distributions1,4. Genotype determination of HCV is one of the most important factors in order to prediction of the viral persistency, pathogenicity and resistancy to antivirals7. The success and the treatment period of interferon and ribavirin seems to be related to the genotype of virus8. Furthermore, HCV genotyping is a useful tool to determine its molecular epidemiology, as they are indicative of transmission route of infection9,10. There is no published data about the distribution of HCV genotypes from Bushehr province (South West of Iran). Prevalence of HCV genotypes in Bushehr is an issue that is not sufficiently investigated and there is a need, therefore, to study this in detail

Using Pharmacokinetic and Viral Kinetic Modeling To Estimate the Antiviral Effectiveness of Telaprevir, Boceprevir, and Pegylated Interferon during Triple Therapy in Treatment-Experienced Hepatitis C Virus-Infected Cirrhotic Patients.: Effectiveness of triple therapy in cirrhotic patients

International audienceTriple therapy combining a protease inhibitor (PI) (telaprevir or boceprevir), pegylated interferon (PEG-IFN), and ribavirin (RBV) has dramatically increased the chance of eradicating hepatitis C virus (HCV). However, the efficacy of this treatment remains suboptimal in cirrhotic treatment-experienced patients. Here, we aimed to better understand the origin of this impaired response by estimating the antiviral effectiveness of each drug. Fifteen HCV genotype 1-infected patients with compensated cirrhosis, who were nonresponders to prior PEG-IFN/RBV therapy, were enrolled in a nonrandomized study. HCV RNA and concentrations of PIs, PEG-IFN, and RBV were frequently assessed in the first 12 weeks of treatment and were analyzed using a pharmacokinetic/viral kinetic model. The two PIs achieved similar levels of molar concentrations (P = 0.5), but there was a significant difference in the 50% effective concentrations (EC50) (P = 0.008), leading to greater effectiveness for telaprevir than for boceprevir in blocking viral production (99.8% versus 99.0%, respectively, P = 0.002). In all patients, the antiviral effectiveness of PEG-IFN was modest (43.4%), and there was no significant contribution of RBV exposure to the total antiviral effectiveness. The second phase of viral decline, which is attributed to the loss rate of infected cells, was slow (0.19 day(-1)) and was higher in patients who subsequently eradicated HCV (P = 0.03). The two PIs achieved high levels of antiviral effectiveness. However, the suboptimal antiviral effectiveness of PEG-IFN/RBV and the low loss of infected cells suggest that a longer treatment duration might be needed in cirrhotic treatment-experienced patients and that a future IFN-free regimen may be particularly beneficial in these patients

Characterization of antibody-mediated neutralization directed against the hypervariable region 1 of hepatitis C virus E2 glycoprotein

The hypervariable region 1 (HVR1) comprising the first 27 aa of E2 glycoprotein is a target for neutralizing antibodies against hepatitis C virus (HCV), but the mechanisms of this neutralization in the cell-culture-infectious genotype 2a strain JFH1 HCV virus (HCVcc) system are unknown. Two rabbit polyclonal sera, R1020 and R140, recognizing the HVR1 of the genotype 1a isolates H77c and Glasgow (Gla), respectively, and a Gla HVR1-specific mouse mAb AP213 have been described previously. However, attempts to generate of antibodies to the JFH1 HVR1 were unsuccessful. Therefore, this study produced chimeric JFH1 HCVcc viruses harbouring the H77c or Gla HVR1 to assess the reactivity of antibodies to this region and their effects on virus infectivity. The inter-genotypic HVR1 swap did not significantly affect virus infectivity. The genotype 1a HVR1-specific antibodies neutralized chimeric viruses in an isolate-dependent manner, underlining the role of HVR1 in HCV infection. The neutralizing antibodies reacted mainly with the C-terminal portion of HVR1, and detailed mapping identified A17, F20 and Q21 in the Gla HVR1 sequence and T21 (and possibly L20) in the corresponding H77c sequence as key epitope residues for AP213 and R140, and R1020, respectively. Importantly, none of the antibodies inhibited in vitro binding of viral envelope glycoproteins to the best-characterized HCV receptor, CD81, or to the glycosaminoglycan attachment factors. However, the HVR1 antibodies were capable of post-attachment neutralization. Overall, this study emphasizes the role of HVR1 in HCVcc entry and provides new tools to study this region further in the context of complete virions

The Hepatitis C Virus Glycan Shield and Evasion of the Humoral Immune Response

Despite the induction of effective immune responses, 80% of hepatitis C virus (HCV)-infected individuals progress from acute to chronic hepatitis. In contrast to the cellular immune response, the role of the humoral immune response in HCV clearance is still subject to debate. Indeed, HCV escapes neutralizing antibodies in chronically infected patients and reinfection has been described in human and chimpanzee. Studies of antibody-mediated HCV neutralization have long been hampered by the lack of cell-culture-derived virus and the absence of a small animal model. However, the development of surrogate models and recent progress in HCV propagation in vitro now enable robust neutralization assays to be performed. These advances are beginning to shed some light on the mechanisms of HCV neutralization. This review summarizes the current state of knowledge of the viral targets of anti-HCV-neutralizing antibodies and the mechanisms that enable HCV to evade the humoral immune response. The recent description of the HCV glycan shield that reduces the immunogenicity of envelope proteins and masks conserved neutralizing epitopes at their surface constitutes the major focus of this review

Analysis of the RNA chaperoning activity of the hepatitis C virus core protein on the conserved 3′X region of the viral genome

The core protein of hepatitis c virus (HCV) is a structural protein with potent RNA chaperoning activities mediated by its hydrophilic N-terminal domain D1, which is thought to play a key role in HCV replication. To further characterize the core chaperoning properties, we studied the interactions between core D1 and the conserved HCV 3′X genomic region required for genome replication. To this end, we monitored the real-time annealing kinetics of native and mutated fluorescently labelled 16-nt palindromic sequence (DLS) and 27-nt Stem Loop II (SL2) from X with their respective complementary sequences. Core D1 and peptides consisting of the core basic domains were found to promote both annealing reactions and partly switch the loop–loop interaction pathway, which predominates in the absence of peptide, towards a pathway involving the stem termini. The chaperone properties of the core D1 peptides were found to be mediated through interaction of their basic clusters with the oligonucleotide phosphate groups, in line with the absence of high affinity site for core on HCV genomic RNA. The core ability to facilitate the interconversion between different RNA structures may explain how this protein regulates RNA structural transitions during HCV replication

Efficacy of octreotide in diarrhoea due to Vibrio cholerae: a randomized, controlled trial

Although octreotide, a long-acting analogue of somatostatin, is currently used in the treatment of chronic secretory diarrhoea due to various causes, its role in the management of acute secretory diarrhoea is not well established. In the present study, therefore, the therapeutic value of octreotide in the management of cholera, a classical example of acute secretory diarrhoea, was investigated. During an outbreak of cholera, patients admitted with acute secretory diarrhoea of \u3c or = 24 h duration and a purging rate \u3e 100 ml/h were enrolled on the study and randomly assigned to octreotide (N = 17) and control (N = 16) groups. All 33 patients received intravenous fluid replacement and antibiotic treatment (200 mg ofloxacin twice daily for 3 days, by mouth). Each patient in the octreotide group was also given a subcutaneous injection containing 100 micrograms octreotide every 8 h for a maximum of six doses. The stool output of each patient was recorded every hour until there had been none for an hour, which was taken as the endpoint. Mean (S.D.) total stool output was lower [6.56 (3.7) v. 9.7 (6.5) litres] and the mean (S.D.) duration of diarrhoea after admission was shorter [32.9 (15.6) v. 47.8 (22.3); P \u3c 0.05] in the octreotide group than in the control group. However, as both groups generally had similar purging rates, the higher volume of stools from the control group was simply the result of the longer period of diarrhoea in this group. Octreotide therefore only decreased the duration of diarrhoea in the cholera patients

A set of reference sequences for the hepatitis C genotypes 4d, 4f, and 4k covering the full open reading frame

Infection with genotype 4 of the Hepatitis C virus is common in Africa and the Mediterranean area, but has also been found at increasing frequencies in injection drug users in Europe and North America. Full length viral sequences to characterize viral diversity and structure have recently become available mostly for subtype 4a, and studies in Egypt and Saudi Arabia, where high proportions of subtype 4a infected patients exist, have begun to establish optimized treatment regimens. However knowledge about other subtype variants of genotype 4 present in less developed African states is lacking. In this study the full coding region from so far poorly characterized variants of HCV genotype 4 was amplified and sequenced using a long range PCR technique. Sequences were analyzed with respect to phylogenetic relationship, possible recombination and prominent sequence characteristics compared to other known HCV strains. We present for the first time two full-length sequences from the HCV genotype 4k, in addition to five strains from HCV genotypes 4d and 4f. Reference sequences for accurate HCV genotyping are required for optimized treatment, and a better knowledge of the global viral sequence diversity is needed to guide vaccines or new drugs effective in the world wide epidemic