Hepatitis C Virus Type Research Articles

Updated Clinical Guidelines on the Management of Hepatitis C Infection in Children.

Children represent only a small proportion of those infected with the hepatitis C virus (HCV) compared to adults. Nevertheless, a substantial number of children have chronic HCV infection and are at risk of complications including cirrhosis, portal hypertension, hepatic decompensation with hepatic encephalopathy, and hepatocellular carcinoma in adulthood. The overall prevalence of the HCV in children was estimated to be 0.87% worldwide. The HCV spreads through the blood. Children born to women with chronic hepatitis C should be evaluated and tested for HCV due to the known risk of infection. The course of treatment for hepatitis C depends on the type of HCV. Currently, there are two pan-genotype HCV treatments (Glecaprevir/pibrentasvir and Sofosbuvir/velpatasvir) for children. We aim to review the updated clinical guidelines on the management of HCV infection in children, including screening, diagnosis, and long-term monitoring, as well as currently published clinical trials and ongoing research on direct acting antiviral hepatitis C treatment in children.

Safety and efficacy of sofosbuvir-velpatasvir to treat chronic hepatitis C virus infection in treatment-naive patients in Rwanda (SHARED-3): a single-arm trial

Hepatitis C virus (HCV) genotype 4 is the predominant type of HCV found in sub-Saharan Africa. Various genotype 4 subtypes, such as 4r, frequently have resistance-associated substitutions that can increase rates of treatment failure with common direct-acting antiviral regimens. In-vitro studies suggest that the NS5A inhibitor velpatasvir is effective against viral isolates containing such resistance-associated substitutions, but its clinical efficacy against genotype 4 non-a/d subtypes in sub-Saharan Africa remains to be confirmed. We aimed to evaluate the safety and efficacy of sofosbuvir-velpatasvir among adults chronically infected with HCV and naive to direct-acting antiviral treatment in Rwanda, where genotype 4 non-a/d subtypes predominate. In this single-arm prospective trial, we enrolled adults (age ≥18 years) in Rwanda who had chronic HCV infection and a plasma HCV RNA titre of at least 1000 IU/mL. Patients were referred from hospitals with HCV treatment programmes throughout Rwanda and were sequentially enrolled and assessed for eligibility at a single study site. Individuals with decompensated liver disease or hepatitis B virus co-infection were excluded. Participants were given an oral fixed-dose combination tablet of sofosbuvir (400 mg) and velpatasvir (100 mg) once-daily for 12 weeks. The primary endpoint was the proportion of participants with a sustained virological response 12 weeks after completion of treatment (SVR12) in the intention-to-treat population. Viral sequencing of the NS5A and NS5B genes was done at baseline for all participants and end of follow-up (week 24) for participants who did not have SVR12. This study is registered with ClinicalTrials.gov (NCT03888729) and is completed. Between Sept 23, 2019, and Jan 10, 2020, 73 individuals were screened for eligibility, of whom 12 (16%) were excluded and 61 (84%) were enrolled. 40 (66%) participants were female, 21 (34%) were male, median age was 64 years (IQR 51-74), and median baseline HCV viral load was 5·7 log10 IU/mL (5·2-6·2). The genotypes identified among the participants were 4k (28 [46%] participants), 4r (11 [18%]), 4v (eight [13%]), 4q (five [8%]), 4l (three [5%]), 4b (one [2%]), 4c (one [2%]), and one undetermined genotype 4 subtype. Three isolates could not be sequenced and were of indeterminate genotype. Of the 55 HCV isolates that were successfully sequenced, all had at least two NS5A resistance-associated substitutions. 59 (97% [95% CI 89-99]) participants had SVR12. 18 (30%) participants had grade 3 adverse events (including 12 [20%] with hypertension), and none had grade 4 adverse events. Four (7%) participants had serious adverse events, including one asthma exacerbation, one abscess, one uterine myoma, and one pelvic fracture related to a motor vehicle accident. No serious adverse events were attributed to the study drug and no adverse event resulted in discontinuation of the study drug. A 12-week regimen of sofosbuvir-velpatasvir is safe and efficacious in treating chronic HCV genotype 4 infection in patients in Rwanda. This regimen could be an effective treatment option in regions known to have a high prevalence of HCV genotype 4 of diverse non-a/d subtypes. Gilead Sciences.

The predominance of Hepatitis C Virus Genotypes and Its Association with the Viral Load in Bangladeshi Population

Determination of hepatitis C virus (HCV) genotype and viral load are two significant prognostic and assessment markers of treatment decisions. The study aimed to determine the predominant HCV types or subtypes and any association with the viral load in Bangladeshi chronic HCV infected patients. A total of 359 anti-HCV positive patients underwent investigation to estimate viral load and determination of genotype and subtype using real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR). Among 306 detectable viral loads containing individuals, 278 (90.85%) genotyped successfully, and 28 (9.15%) had unknown genotypes. Among typable genotypes, 1a accounted for 14 (5.03%), 1b for 14 (5.03%), 3 for 247 (88.85%), 4 for 2 (0.72%) and genotype 6 for 1 (0.36%). Based on pre-treatment viral load levels, study subjects classified into three categories such as low (<50000 IU/mL), intermediate (50000-500000 IU/mL), and high (>500000 IU/mL). The majority of HCV other types (1a, 1b, 4, 6) infected patients (96.4%) had intermediate to high viral load compared to those infected with genotype 3 (77.7%) and unclassified types (55.0%) (χ2 =15.41; p = 0.004). HCV type 3 was prevalent (68.4%) in the above 40 years of group compared to less than 40 years group (31.6%). HCV genotype 3 was the predominant genotype circulating in Bangladesh. Pre-treatment viral load demonstrated significant difference among individuals having HCV other types and type 3. However, sequencing the HCV genome analysis would determine the exact types and subtypes among all possible HCV strains available in Bangladesh.
 Bioresearch Commu. 7(1): 955-959, 2021 (January)

Detection of circulating HCV recombinant form RF1_2k/1b in blood serum of patients by real-time RT-PCR.

Globally, about 70 million people are infected with the hepatitis C virus (HCV), and about 400 thousand people die annually from chronic hepatitis C complications. The management of patients with chronic hepatitis C may require HCV genotyping, since the efficiency of some widely used antiviral drugs strongly depend on the viral genotype and/or subtype. The most prevalent HCV circulating recombinant form, RF1_2k/1b, is misclassified as genotype 2 by many commercial HCV genotyping kits, based on the RT-PCR analysis of the 5' untranslated region of the HCV genome. This leads to inappropriate patient treatment, since the accepted treatment schemes for HCV genotype 2 are ineffective for the RF1_2k/1b. Here we describe a method for detecting the RNA HCV RF1_2k/1b in blood samples by RT-PCR analysis of two regions in HCV genome (5'UTR and NS5b). The method was tested on 240 blood serum samples from HCV infected patients, in which HCV genotype was defined as 2 or mixed (2+1 or 2+3) by the two commercial genotyping kits "OT-Hepatogen-C genotype" ("DNA-Technology", Moscow) and "RealBest RNA HCV-1/2/3" ("Vector- Best ", Novosibirsk). 50 (20.8%) RF1_2k/1b cases were revealed, including three mixed infections: RF1_2k/1b + 1a, RF1_2k/1b + 3a, RF1_2k/1b + 1b. In all cases, the accuracy of HCV typing by the proposed method was confirmed by Sanger sequencing and phylogenetic analysis. The method is easy to implement into clinical practice and may be used in clinical settings equipped for RT-PCR analysis to correctly identify the recombinant variant RF1_2k/1b.

COVID-19 Among Patients With Hepatitis B or Hepatitis C: A Systematic Review

Context: Hepatic manifestations of Coronavirus Disease 2019 (COVID-19) are common among people living with Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV). Objectives: This systematic review aimed to summarize the evidence on COVID-19 patients living with HBV or HCV co-infections. Data Sources: We searched multiple electronic databases and preprint servers from December 1, 2019, to August 9, 2020. Study Selection: Studies were included if they reported quantitative empirical data on COVID-19 patients living with HBV or HCV co-infections. Data Extraction: Descriptive analyses were reported, and data were synthesized narratively. The quality assessment was completed using the Joanna Briggs Institute critical appraisal tools. Results: Out of the 941 uniquely identified records, 27 studies were included. Of the eligible studies, 232 COVID-19 patients were living with HBV and 22 were living with HCV. Most patients were male, and the mean age was 49.8 and 62.8 years in patients living with HBV and HCV, respectively. Among the reported cases of SARS-CoV-2-HBV co-infection, the proportions of death were 4.7% and 15% in cross-sectional and case series/report studies, respectively. The death proportion was 8.3% among the reported cases of SARS-CoV-2-HCV co-infection. Among COVID-19 patients, 34.1% and 76.2% reported at least one comorbidity besides HBV and HCV infections, mainly hypertension and type 2 diabetes mellitus. The most common COVID-19-related symptoms in both HBV and HCV groups were fever, cough, dyspnea, fatigue, and gastrointestinal symptoms. Conclusions: While understanding the pathogenesis of SARS-CoV-2 requires further investigations, the careful assessment of hepatic manifestations and chronic infections, such as HBV and HCV upon the admission of COVID-19 patients could help reduce multimorbidity among HBV or HCV patients and lead to more favorable health outcomes among them.

Post-transplantation diabetes mellitus after kidney transplantation and its related factors in Iranian patients: a retrospective single-center study

Introduction: Post-transplantation diabetes mellitus (PTDM) is a metabolic complication following transplantation, which is associated with cardiovascular disease and leads to increased post-ttransplantation morbidity and mortality. Objectives: To identify the incidence of PTDM and its risk factors in kidney recipients at a single-center in Iran. Patients and Methods: This retrospective study was conducted on 379 kidney recipients with a negative history of diabetes mellitus who underwent transplant before January 2017. PTDM was defined according to the diagnostic criteria of the American Diabetes Association (ADA) and the World Health Organization (WHO). Data on demographic, clinical characteristics and laboratory parameters were collected. Kaplan-Mayer analysis was used to evaluate the cumulative incidence of PTDM. The association between risk factors and PTDM incidence was identified with stepwise Cox regression. Results: The cumulative incidence of PTDM during a 24-month follow-up was 30.1% (95% CI: 25.6-34.8). By univariate analysis, modifiable or non-modifiable risk factors for PTDM development included recipient age, body mass index (BMI), marital status, family history of diabetes, smoking, type of transplant, hepatitis C virus (HCV), cytomegalovirus (CMV), transplant rejection, TG, tacrolimus, cyclosporine and beta blocker. In this study, family history of diabetes, type of transplant, HCV, CMV, TG, tacrolimus, and beta blocker were predictors of development of PTDM in Cox proportional hazard models. Conclusion: The incidence of PTDM was high. Identification of risk factors determines appropriate strategies for PTDM incidence risk reduction.

Changes on Hepatitis C virus genotype distribution in Western Turkey: Evaluation of twelve-year data.

Hepatitis C virus (HCV) prevalence is 1% in Turkey with genotype 1 being the predominant type traditionally. However unique geographical location of Turkey and increasing human migration in the region influences the epidemiology of the infection. The aim of this study was to determine the changes in distribution of HCV genotypes and risk factors. In this retrospective single-center study, HCV genotyping results of 558 patients were evaluated in between 2005 and 2016.Three different HCV genotyping assays were used during the 12-year study period;restriction fragment length polymorphism (RFLP), Abbott Real Time HCV Genotype II and Bosphore HCV genotyping kit. The most prevalent HCV genotype was genotype 1 detected in 88.4% of the patients followed by genotype 3 (5.2%),genotype 4 (2.9%),genotype 2 (2.1%), mixed genotypes (1.1%) and genotype 5 (0.3%).Genotype 1a showed an increasing prevalence.There were 19 patients (3.4%) either of foreign nationalities or Turkish citizens living abroad. Genotype 3 was the most common type among these patients which 10.3% had intravenous drug use history.Syrian migrant population differed in terms of HCV genotypes.Genotype 5 detected in two Syrian patients, which is the first report of HCV type 5 in Western Turkey. Among the HCV genotype 4 infected patients, 31.3% were Syrians. Our study showed that although genotype 1b dominance continues, the distribution and prevalence of HCV genotypes are changing in our region mainly due to migration and increase in the frequency of patients with non-traditional risk factors such as intravenous drug use. Monitoring the epidemiology of HCV genotypes may provide guidance in treatment decisions.

2197. Hepatitis B Reactivation in Patients with Malignancies Undergoing Treatment for Hepatitis C Infection with Direct-Acting Antivirals

BackgroundReactivation of hepatitis B virus (HBV) can occur in patients after cancer therapies. Direct-acting antivirals (DAAs) are the effective therapies for hepatitis C virus (HCV) infection, and HBV reactivation in HCV/HBV co-infected patients treated with DAAs has been reported. We analyzed the risk of HBV reactivation among HCV/HBV co-infected cancer patients being treated with DAAs.MethodsWe prospectively followed patients with any type of cancer and HCV treated with DAAs between January 2014 and January 2018 at MD Anderson Cancer Center. Information on demographics, use of radiation, chemotherapy, immunotherapy, or anti-CD20 antibodies, and anti-HBV therapy were collected. All patients had the following tests at baseline, 2 and 4 weeks after initiation of DAAs, at end of treatment (EOT), and 12 weeks after completion of DAAs: alanine aminotransferase, total bilirubin, HBV surface antigen (HBsAg), hepatitis B core antibody (anti-HBc), and HBV DNA and HCV RNA levels. We defined the following outcomes by AASLD-recommended parameters: HBV reactivation (HBsAg reverse seroconversion, HBV DNA >2 log compared with baseline, HBV DNA >3 log if HBV DNA was undetectable, or >4 log if baseline was unavailable), hepatitis flare (ALT increase ≥3 times baseline and >100 U/L), and HBV-associated hepatitis (HBV reactivation and hepatitis flare). Patients were followed for 12 weeks after completion of DAAs.ResultsOf 169 cancer patients treated for HCV infection, 2.4% (n = 4) had chronic HBV infection (HBsAg+/anti-HBc+), and most (3/4) of these were on anti-HBV therapy. Past HBV infection (HBsAg-/HBcAb+) was noted in 30% (51/166), and none received anti-HBV therapy. Of these, 37% (19/51) had cancer therapy within 6 months prior to DAA treatment. HBV reactivation did not occur in any co-infected patients. Two patients had hepatitis flare, but none developed HBV-associated hepatitis.ConclusionThis is the first prospective study evaluating HBV reactivation in HCV/HBV co-infected cancer patients receiving DAAs. The risk of HBV reactivation in these patients seems to be low. Future studies with a larger cohort of co-infected cancer patients allowing personalized risk stratification are needed.Disclosures J. P. Hwang, Gilead Sciences: Investigator, Grant recipient; Merck & Co., Inc.: Investigator, Grant recipient. H. Torres, Gilead Sciences, Merck & Co., Inc.: Grant Investigator, Grant recipient; Vertex Pharmaceuticals: Grant Investigator, Grant recipient.

2228. Late Viral Relapse After Direct-Acting Antiviral Treatment in Hepatitis C Virus-Infected Cancer Patients

BackgroundAccording to professional societies, the endpoint to consider hepatitis c virus (HCV) infection cured is the achievement of a sustained virologic response 12 weeks after treatment completion (SVR12). Late recurrences (beyond SVR12) are rare. Herein, we report two cases of HCV-infected cancer patients with late relapses post direct-acting antivirals (DAAs).MethodsPatients with any type of chronic cancer and HCV treated with DAAs between January 2014 and March 2018 at MD Anderson Cancer Center were prospectively followed. All patients had HCV RNA levels at baseline; 2 and 4 weeks after initiation of DAAs; at end of treatment (EOT); and 12 weeks after completion of DAAs. No phylogenetic analyses were available for samples collected.ResultsAmong 196 HCV-infected cancer patients treated with DAAs, 20 developed viral relapse, 2 (10%) of them with late relapse (Figure 1). Both patients denied behaviors, exposures, and conditions associated with HCV reinfection. Case 1: Fifty-six-year-old male with hepatocellular carcinoma (HCC), HCV genotype 1a, interferon-experienced, with compensated cirrhosis received in 2017 ledipasvir/sofosbuvir for 12 weeks, followed by systemic chemotherapy with sorafenib. He achieved an SVR12 but developed HCV relapse 12 weeks later (24 weeks after EOT). Patient remained infected with HCV 1a. He did not receive retreatment due to HCC not amenable for curative treatment. Case 2: 57-year-old male with multiple myeloma, HCV genotype 1a, interferon-experienced without cirrhosis. He received sofosbuvir and simeprevir in 2015 for 12 weeks. Post DAAs, he received chemotherapy with carfilzomib, lenalidomide, dexamethasone, and ixazomib followed by autologous hematopoietic cell transplant pre-conditioned with melphalan. He achieved both an SVR12 and SVR 24 but had HCV relapse detected during the one year follow-up visit. Patient remained infected with HCV 1a. He has retreated with sofosbuvir, veltapasvir, voxilaprevir and ribavirin and currently with HCV RNA level at EOT.ConclusionLate HCV relapses can occur in HCV-infected cancer patients. Long-term monitoring of HCV-RNA and easy-to-use tests to differentiate relapses from reinfection in real-world practice are warranted in this population. Disclosures H. Torres, Gilead Sciences, Merck & Co., Inc.: Grant Investigator, Grant recipient. Vertex Pharmaceuticals: Grant Investigator, Grant recipient.

Real Time PCR Based Detection of Hepatitis C Virus Major Genotypes in Chronic HCV Patients in Khyber Pakhtunkhwa

Background: Hepatitis C virus (HCV) has been classified into seven major and more than 100 quasi species. To treat hepatitis C patients, it is important to determine the type of HCV as it has been linked with treatment duration and outcome. Objectives: The objective of the study was to find out HCV major genotypes using Taqman Probe based real time PCR assay and to compare prevalence of these with previous HCV genotype distribution using other methods in Khyber Pakhtunkhwa. Methods: In this study, 100 actively infected patients were selected, referred by different physicians, and genotyping was done using Sacace real time PCR based assay. Results: Among 100 patients, the most abundant major genotype was HCV 3, followed by genotype 1 and the untypable while genotype 2 was scarcely found. Conclusions: The existing methodology confirmed the prevalence of major genotype 3, similar to that of previous methodologies and studies, however, with a slight shift of genotype 2 distribution.

IL-28B rs12979860 polymorphism affect the course of chronic hepatitis and the development of HCC in Egyptian patients with hepatitis C type 4

ABSTRACTBackground: A single nucleotide polymorphism (SNP) in the interleukin 28B (IL28B) gene may alter the trajectory of hepatitis C virus (HCV) chronic infection. Several studies have sought to determine a link between IL28B rs12979860 SNP and the development of HCV-related hepatocellular carcinoma (HCC), but with variable results, and consensus is awaited. We hypothesised that IL28B rs12979860 SNP is linked to HCC in patients with HCV type 4.Methods: IL28B genotyping of 300 patients with HCV-related fibrosis (n = 100), cirrhosis (n = 100) and HCC (n = 100) was carried out and the results were analysed to determine the association between the IL28B genotype and clinical outcome.Results: In IL28B TT genotype carriers, the proportions of moderate/severe fibrosis, advanced cirrhosis (Child B-C) and HCC (50%, 84% and 60.2%, respectively) were higher (p < 0.05) than in CC/CT (4.3%, 46% and 23%, respectively). IL-28B SNP was linked significantly (p < 0.05) with cirrhosis progression and HCC advanced stages. Moreover, HCC advanced Child, Okuda and CLIP stages were associated with T allele carriage (73.9%, 82.6% and 78.3% vs. 44.2%, 50.6% and 46.8% in CC/CT). The percentage of large tumour size (> 3cm) increased (p = 0.028) in TT genotype carriers (81.8% vs.52.6% in CC/CT).Conclusion: IL-28B rs12979860 TT genotype is more prevalent in patients with advanced fibrosis, cirrhosis and HCC stages. Thus, it seems to be associated with poor outcomes in chronic HCV patients and to augment the risk of developing HCC.

Structural and Epitope Analysis (T- and B-Cell Epitopes) of Hepatitis C Virus (HCV) Glycoproteins: An in silico Approach

Chronic infection with Hepatitis C Virus (HCV) poses a major risk for liver disease like cirrhosis, liver failure and hepatocellular carcinoma. In terms of percentage, the prevalence of HCV in India was found to be low to moderate (1-1.5%), but in terms of sheer numbers, India has a significant number of global HCV patients. Presently, HCV can be treated with direct acting-antibody drugs but there is no prophylactic or therapeutic vaccine available against it. In HCV infection, T- and B-cell immunity is important for clearing the virus. In the present study immunoinformatics was used to identify potent vaccine target for HCV vaccine development. Sequence of HCV was retrieved from NCBI and their structural analysis was done by using Protpram, PSIPRED, iTASSER and PDBsum servers. T-cell and B-cell epitopes were predicted by Immune Epitope Database and ACBPRED servers. On epitope prediction, 25 and 55 potent MHC-I epitopes and 7 and 13 potent B-cell epitopes were predicted for E1 and E2 protein respectively. Their antigenicity score was also calculated. The most potent MHC-I epitopes were MMMNWSPAV and MAWDMMMNW for HLA-A*02:01 and HLA-B*53:01 and most potent B-cell epitope was TGHRMAWDMMMNWSPA for E1 protein. For E2, four MHC-I epitopes having the lowest binding energy and most potent B-cell epitope was DRPYCWHYAPRPCDTI. In the present study, most potent epitopes for HCV was determined on the basis of their antigenicity along with 3D modeling and docking. Identified B- and T-cell epitopes can be used for the development of potent vaccine against most prevalent HCV type in India to limit its infection.

Intergenotypic 2k/1b hepatitis C virus recombinants in the East Macedonia and Thrace region of Greece.

BackgroundIntergenotypic recombinant hepatitis C virus (HCV) strains emerge rarely during coinfection of the same individual with two HCV genotypes. Few recombinant HCV strains have been identified to date and only one, CRF01 2k/1b, has become a worldwide concern. This study reevaluated the genotyping of three HCV genotype 2 strains from a group of patients with an unusually low rate of sustained virological response after pegylated interferon/ribavirin treatment. In addition, genetic determinants of host interferon resistance were evaluated.MethodsThe HCV type 2 strains from the patients’ serum were subjected to partial sequencing of the core-E1, NS2, NS5A and NS5B regions by reverse transcription polymerase chain reaction. Furthermore, the IFNL3 rs12979860 and the IFNL4 rs368234815 single nucleotide polymorphisms were defined in two of the three patients.ResultsAll three strains were phylogenetically related to the Russia-derived CRF01 2k/1b while they encompassed the exact same 2k/1b junction site within NS2.ConclusionThis is the first report of HCV 2k/1b recombinants in Greece and the greater area of the Balkans.

MxA is a positive regulator of type I IFN signaling in HCV infection.

Type I interferons (IFNs) are a family of primordial cytokines that respond to various pathogen infections including Hepatitis C virus (HCV). Type I IFNs signal through Jak/STAT pathway leading to the production of a few hundred interferon stimulated genes (ISGs). The aim of this study was to explore the role of one of these ISGs, MxA in HCV infection and type I IFN production. Plasmid encoding MxA was cloned into PcDNA3.1-3×tag vector and MxA expression was confirmed both at mRNA (RT-PCR) and protein (Western blot, WB) levels. IFNα and IFNβ productions were quantified by RT-PCR from cell lysate and by ELISA kit from culture medium following MxA over-expression in Huh7.5.1 cells. The activation status of Jak/STAT signaling pathway was examined at three levels: p-STAT1 (WB), interferon sensitive response element (ISRE) activity (dual luciferase reporter gene assay), and levels of ISG expression (RT-qPCR). J6/JFH1 HCV culture system was used to study the role of MxA in HCV replication. Our findings indicated that MxA over-expression inhibited HCV replication and potentiated the IFNα-mediated anti-HCV activity; MxA stimulated the production of IFNα, IFNβ, and enhanced IFNα-induced activation of Jak-STAT signaling pathway. We concluded that MxA is a positive regulator of type I IFN signaling in HCV infection.

A Novel Human Pegivirus, HPgV-2 (HHpgV-1), Is Tightly Associated With Hepatitis C Virus (HCV) Infection and HCV/Human Immunodeficiency Virus Type 1 Coinfection

Human pegivirus type 2 (HPgV-2) is a novel blood-borne human pegivirus that mainly infects hepatitis C virus (HCV)-infected subjects. We have investigated the prevalence of HPgV-2 in China, its association with HCV and human immunodeficiency virus type 1 (HIV-1), and the impact on HCV viral load and liver damage. A cross-sectional study was conducted with both blood donors and HCV- and HIV-1-infected patients in Guangzhou, China. All subjects were screened for anti-HPgV-2 and HPgV-2 RNA. Demographic and clinical information were obtained from electronic medical records. We tested 8198 serum or plasma samples. Only 0.15% (6/4017) of healthy blood donors were positive for anti-HPgV-2 and negative for HPgV-2 RNA. No HPgV-2 viremia was detected in hepatitis B virus- or HIV-1-monoinfected individuals. The relatively high frequency of HPgV-2 infection was observed in 1.23% (30/2440) and 0.29% (7/2440) of HCV-infected persons by serological assay and reverse-transcription polymerase chain reaction, respectively. Furthermore, anti-HPgV-2 and HPgV-2 RNA were detected in 8.91% (18/202) and 3.47% (7/202), respectively, of HCV/HIV-1-coinfected subjects. HPgV-2 persistent infection was documented in about 30% of anti-HPgV-2-positive individuals. In addition, HPgV-2 infection may not affect HCV-related liver injury and HCV viral load. Our results indicate the rarity of HPgV-2 infection in the general population and tight association with HCV, in particular with HCV/HIV-1 coinfection. HPgV-2 appears not to worsen HCV-related liver damage. Our study provides new findings about the association of HPgV-2 and HCV/HIV-1 and the impact of HPgV-2 infection on HCV replication and pathogenesis.

Identification of Genotype 2 HCV in Serotype-1 Hepatitis C Patients Unresponsive to Daclatasvir plus Asunaprevir Treatment.

It is important to determine the genotypes or serotypes of hepatitis C virus (HCV) in patients before treatment with direct-acting antiviral agents (DAAs), because the effects of DAAs differ among genotypes. In Japan, two tests for HCV typing are available clinically, but only serotyping, not genotyping, is approved by the public health insurance. Although most serotype-1 Japanese patients are infected with genotype 1b HCV, it is known that a small proportion of patients show different results from two typing methods. This study focused on such patients and the effectiveness of treatment with daclatasvir plus asunaprevir (DCV/ASV) was evaluated. We analyzed 644 DCV/ASV-treated patients with serotype 1 or genotype 1b, and among them, 166 serotype-1 patients received a commercial-based direct sequencing (DS) test for resistant-associated variants of genotype 1b HCV. We found four patients (2.4%) with DS test failure, suggesting that the PCR primers targeting genotype 1b may not match. Importantly, none of the four patients achieved a sustained virological response. Our in-house DS test analyzing the 5'-untranslated region and coding regions for NS4 and NS5B of HCV showed that three of the four patients were infected with genotype 2 HCV, and one patient was infected with genotype 1a HCV. No recombinant virus of different genotypes was found. This study indicates that a subset of serotype-1 hepatitis C patients is infected with HCV of genotype 2 or 1a in Japan and that DCV/ASV is not effective for such patients. Thus, attention should be paid to DAA treatment without HCV genotyping.

Hepatitis C virus E2 protein encapsulation into poly d, l-lactic-co-glycolide microspheres could induce mice cytotoxic T-cell response.

Hepatitis C virus (HCV) is known to cause hepatitis and hepatocellular carcinoma. E2 envelope glycoprotein of HCV type (HCV-E2) has been reported to bind human host cells and is a major target for developing anti-HCV vaccines. However, the therapeutic vaccine for infected patients still needs further development. The vaccine aims to provide cytotoxic T-cells to eliminate infected cells and hepatocellular carcinoma. Currently, there is no effective HCV therapeutic vaccine because most chronically infected patients rarely generate cytotoxic T-cells, even though they have high levels of neutralizing antibodies. Therefore, the adjuvant must be applied to enhance the efficacy of the therapeutic vaccine. In this study, we constructed HCV1b-E2 recombinant protein, a truncated form of peptide, to combine with an effective vaccine adjuvant and delivery system by using poly d,l-lactic-co-glycolide (PLGA) microspheres. HCV1b-E2 protein was effectively encapsulated into PLGA microspheres (HCV1b-E2-PLGA) as a strategy to deliver an insoluble form of HCV1b-E2 protein. The size and shape of PLGA microspheres were generated properly to carry an insoluble form of viral peptide in vivo. The encapsulated viral protein was slowly and continuously released from PLGA microspheres, which indicated the property of the adjuvant. HCV1b-E2-PLGA can trigger a cell-mediated immune response by inducing an expression of mice CD8+ T-cells. Our results demonstrated that HCV1b-E2-PLGA-immunized mice have a significantly increased CD8+ T-cell number, whereas HCV1b-E2-immunized mice have a lower number of CD8+ T-cells. Moreover, HCV1b-E2-PLGA could induce a specific antibody to viral protein, and the immune cells could secrete IFN-γ, which is a significant cytokine for viral response. Thus, HCV1b-E2-PLGA is shown to have adjuvant property and efficacy in the murine model, which is a good strategy to develop HCV prophylactic and therapeutic vaccines.

New Drug Targets All 6 HCV Types

Joining the wave of recently approved hepatitis C virus (HCV) drugs that act directly on the virus is a tablet containing sofosbuvir, which was approved in 2013, and velpatasvir—the first drug to treat all 6 major forms of HCV infection (http://bit.ly/290Hwgp).

Antiretroviral therapy does not affect response to chronic hepatitis C therapy in HIV-coinfected patients.

Many patients coinfected with the human immunodeficiency virus (HIV) and hepatitis C virus (HCV) are using highly active antiretroviral therapy (HAART) and HCV therapy with peginterferon (PEG-IFN) and ribavirina (RBV) because the use of direct-acting antivirals is not a reality in some countries. To know the impact of such medications in the sustained virological response (SVR) during HCV treatment is of great importance. This was a retrospective cohort study of 215 coinfected HIV/HCV patients. The patients were treated with PEG-IFN and RBV between 2007 and 2013 and analyzed by intention to treat. Treatment-experienced patients to HCV and carriers of hepatitis B were excluded. Demographic data (gender, age), mode of infection, HCV genotype, HCV viral load, hepatic fibrosis, HIV status, and type of PEG were evaluated. One hundred eighty-eight (87.4%) patients were using HAART. SVR was achieved in 55 (29.3%) patients using HAART and in 9 (33.3%) patients not using HAART (p = 0.86). There was no difference in SVR between different HAART medications and regimens using two reverse transcriptase inhibitor nucleosides (NRTIs) or the use of protease inhibitors and non-NRTIs (27.1% versus 31.5%; p = 0.61). The predictive factors for obtaining SVR were low HCV viral load, non-1 genotype, and the use of peginterferon-α2a. The use of HAART does not influence the SVR of HCV under PEG-IFN and RBV therapy in HIV/HCV coinfected patients.

Hepatitis C viral dynamics during ribavirin priming differ according to prior treatment response and HCV type

The mode of action of ribavirin is not completely understood. Ribavirin monotherapy has a measurable antiviral effect, which shows great variability. It might lead to an earlier steady state of plasma concentration and therefore enhance the effect of following combination treatment. The aim of this study was to evaluate the antiviral effect of ribavirin priming and its influence on sustained virologic response after combination treatment in a group of patients with different hepatitis C virus (HCV) types with or without prior treatment experience. Retrospective analysis of 75 patients (37 treatment naïve, 20 prior relapse, 16 prior nonresponse, genotype 1 present in 60 patients) from five centres who received ribavirin priming as part of an individual strategy in order to improve treatment outcome. All patients received ribavirin monotherapy with a mean dose of 14.5mgkg-1 body weight for a mean of 28days. After ribavirin priming, dual combination treatment with pegylated interferon alfa and ribavirin was started. The mean HCV RNA decline after ribavirin priming was 0.6 log10 IUmL-1 (P<.001). The initial viral decline depended on HCV type and previous treatment status being highest among prior relapsers (0.8 log10 IUmL-1 ; P=.002) and HCV type 2/3 (1.2 log10 IUmL-1 ; P=.05) and lowest among those with prior nonresponse (0.3 log10 IUmL-1 , P=.01). IFNL4 (formerly IL28B) genotype for rs12979860 and IFNL3 genotype rs8099917 did not influence the initial viral decline. The study demonstrates a significant variability in the viral dynamics and antiviral efficacy of ribavirin monotherapy, which is mainly influenced by prior treatment status. The fact that the lowest response pattern was observed in prior nonresponder patients to pegylated interferon alfa plus ribavirin combination therapy can be taken as a hint that not only the individual interferon, but also the ribavirin sensitivity contributes significantly to the nonresponsive state.