Hepatitis Delta Virus

The kinetic characteristics have been studied for noncircularly permuted variants of the human hepatitis delta virus antigenomic ribozyme to find out the cause of the two-phase kinetics of the self-cleavage reaction. Different ways of reaction initiation, suboptimal conditions, and jumpwise changes of reaction conditions have been used, and the temperature dependences have been studied. A correlation has been shown between

COS7 cells and other cell lines expressing the SV40 large T-antigen, have been of great benefit in transfection studies using transient expression vectors which contain the SV40 origin of replication, as these cells allow plasmid replication to a high copy number. As no T-antigen expressing cell line derived from a well characterised hepatocyte-like continuous cell line currently exists, the establishment of such a cell line for studies which require expression of hepatocyte-specific factors would be extremely useful. A HepG2-derived stable cell line (THT1) was therefore developed which demonstrates a high level of transfection efficiency whilst retaining hepatocyte-like features, such as the production of hepatitis B virus. The THT1 cell line displayed chromosomal integration of the SV40 T-antigen gene and nuclear expression of the antigen. The cell line also maintained the general morphological features of its parent cell line, and showed an increased rate of cell growth.

Bangladesh is a densely populated country where about 10 million people are chronically infected with hepatitis B virus (HBV). The aim of the present study was to evaluate the biochemical, virological and histological characteristics of HBeAg-negative chronic hepatitis B (CHB). Patients were included in this study if they were chronically infected with HBV with detectable DNA. The patients who were co-infected with human immunodeficiency virus, hepatitis delta virus or hepatitis C virus, and previously subjected to antiviral treatment, and those with hepatocellular carcinoma were excluded. The study was conducted during the period of January 2001 to December 2007. During this period 2617 patients with CHB were studied. HBeAg-positive cases were included to compare the characteristics. Among them, 237 cases underwent liver biopsy. 2296 patients (87.7%) were male, with a mean age of 28.9+/-13.7 years. 2375 patients (90.8%) had CHB, and 242 (9.2%) were cirrhotic. HBV DNA levels were 7....

Metode Pengasapan untuk Mengatasi dan atau Bekerja Efektif membunuh Senjata Biologi Kuman Bakteri Virus Jamur, digunakan untuk membunuh dan atau memperlambat  Penyebaran Kuman, Biological Weapon (Semisal: Senjata Biologi : Corona versi asli, varian Alpha, Beta, Gamma, Delta, dan atau Omicron dan atau Turunannya) dan atau mikroorganisme  pertumbuhan yang keluar melalui Pori Pori Tubuh Manusia dan atau pada Permukaan Kulit Tubuh Manusia, serta menghambat  Penyebaran Kanker,  Penyakit Kulit

The majority of patients with acute febrile jaundice (>95%) identified through a yellow fever surveillance program in the Democratic Republic of Congo (DRC) test negative for antibodies against yellow fever virus. However, no etiological investigation has ever been carried out on these patients. Here, we tested for hepatitis A (HAV), B (HBV), C (HCV), D (HDV) and E (HEV) viruses, all of which can cause acute febrile jaundice, in patients included in the yellow fever surveillance program in the DRC. On a total of 498 serum samples collected from suspected cases of yellow fever from January 2003 to January 2012, ELISA techniques were used to screen for antibodies against HAV (IgM) and HEV (IgM) and for antigens and antibodies against HBV (HBsAg and anti-HBc IgM), HCV and HDV. Viral loads and genotypes were determined for HBV and HVD. Viral hepatitis serological markers were diagnosed in 218 (43.7%) patients. Seroprevalence was 16.7% for HAV, 24.6% HBV, 2.3% HCV and 10.4% for HEV an...

Chronic hepatitis B and D infections are major causes of liver disease and hepatocellular carcinoma worldwide. Efficient therapeutic approaches for cure are absent. Sharing the same envelope proteins, hepatitis B virus (HBV) and hepatitis D virus (HDV) use the sodium taurocholate co-transporting polypeptide (NTCP, a bile acid transporter) as a receptor to enter hepatocytes. However, the detailed mechanisms of the viral entry process are still poorly understood. Here, we established a high-throughput infectious cell culture model enabling functional genomics of HDV entry and infection. Using a targeted RNAi entry screen we identified glypican 5 (GPC5) as a common host cell entry factor for HBV and HDV. These findings advance our understanding of virus cell entry and open new avenues for curative therapies. Since glypicans have been shown to play a role in the control of cell division and growth regulation, virus-GPC5 interactions may also play a role in the pathogenesis of virus-indu...

No specific therapy exists for hepatitis delta virus (HDV), which can cause severe liver disease. Molecular genetic studies have implicated the prenylation site of large delta antigen as a critical determinant of HDV particle assembly. We have established a cell culture model which produces HDV-like particles, and we show that delta antigen prenylation can be pharmacologically inhibited by the prenylation inhibitor BZA-5B. Furthermore, BZA-5B specifically abolishes particle production in a dose-dependent manner. These results demonstrate that the use of such a prenylation inhibitor-based antiviral therapy may be feasible and identify a novel class of potential antiviral agents.

In this study, antibodies against blood-borne hepatitis viruses (Hepatitis B, C and D) were investigated by ELISA in 43 patients with chronic renal insufficiency, at Hemodialysis Unit of Ondokuz Mayis University, Faculty of Medicine. 32 (76.2%) of the patients were found positive Anti-HBc total antibody and 12 (27.5%) patients were HBsAg carriers. 14 (32.5%) patients had Anti-HBs which showed protective immunity. Anti-HBs antibody was found negative in 6 patients after Hepatitis B infection. 9 (75%) of the HBsAg carriers had Anti-Delta antibody. 34 (79.1%) patients showed Anti-HCV positivity, 6 (18.2%) of HCV seropositive patients were found to be positive for HBsAg. There wasn't any correlation between length of time on hemodialysis, age and seropositivity of hepatitis markers. A correlation was detected between multiple transfusion and Anti-HCV seropositivity. The prevalence of HBsAg, Anti-HBc total and Anti-HCV were sequentially found 6%, 38%, 1% among healthy blood donors taken as a control group. Hepatitis virus infection was frequently seen in hemodialysis patients because of multiple blood transfusion. Chronicity is high because of insufficiency of cellular and humoral immunity. Testing blood for hepatitis viruses markers before blood transfusion and Hepatitis B vaccination programme might control viral hepatitis infection in hemodialysis units.

Coinfection of hepatitis B virus (HBV) with hepatitis D virus (HDV) has being reported to increase severity of progression to hepatocellular carcinoma (HCC) and liver cirrhosis (LC). Beta microglobulin (2βM) which is present on the surfaces of blood cells in acceptable levels is a tumor marker which may become elevated in disease conditions. This study hence observed the prevalence of HBV and HDV coinfection in a rural population and their 2βM concentration. Of the 368 samples, 66 (17.9%) were positive to hepatitis B surface antigen (HBsAg) and 33 (50%) were coinfected with HDV, 8 (2.1%) were monoinfected with HDV. 2βM concentration increased beyond the normal level in individuals coinfected with HBV and HDV as compared with the monoinfected individuals. Coinfection resulted in the increased concentration of 2βM in HBV and HDV coinfection and the likelihood of progression to HCC and LC may not be ruled out. Monoinfection with HDV also had high 2βM concentration but this is due to ha...

Patients with dual hepatitis B (HBV) and hepatitis D (HDV) virus infection are at an increased risk of progression to liver cirrhosis and hepatocellular carcinoma than patients with a single viral infection. Treatment of viral hepatitis due to dual HBV/HDV infection represents a challenge. Currently there is no vaccine against HDV. Recombinant production of HDV antigen (HDAg) is the first step towards a potential vaccine candidate and the development of assays for HDV detection. This study demonstrates the expression of one HDAg isoform, S-HDAg, in Pichia pastoris. A recombinant vector carrying a tagged gene encoding S-HDAg under the control of the methanol-inducible promoter AOX1 was designed and integrated into P. pastoris X33. The protein, which was purified using a Ni(2+) affinity column and eluted at 100-150 mM imidazole, has potential as a recombinant antigen for further study.

Epidemiological data obtained to date show that the delta agent is spread all over the world. In Rumania hepatitis B virus infection is widespread and is associated with a high incidence of chronic liver disease. To determine the prevalence of delta infection, sera from 373 patients with chronic liver disease were tested for HBV markers; 228 were HBsAg positive. Anti-HD was present in 190 sera (83.33%), which reveals a very high incidence and shows a higher frequency of the active forms of the disease. However, in contrast to other studies, we found a high incidence of HDV infection in chronic persistent hepatitis and hepatocellular carcinoma.

Summary.  Long-term changes in the frequency and outcome of hepatitis delta virus (HDV) infection have seldom been analysed. This retrospective, longitudinal study includes 398 consecutive hepatitis B surface antigen (HBsAg)-positive patients with anti-HDV antibodies who attended our institution between 1983 and 2008. At enrolment, 182 patients had acute and 216 chronic hepatitis. Patients were grouped into two periods. Those who attended between 1983 and 1995 and those between 1996 and 2008. The former group was significantly younger, mainly intravenous drugs users, and had a greater incidence of acute HDV and HIV and HCV coinfection. Patients with acute HBV/HDV coinfection cleared both infections in 90% of cases, while all patients with HDV superinfection evolved to chronic disease. One hundred and fifty-eight patients with chronic HDV were followed for a median period of 158 months. Seventy-two per cent of the patients remained stable, 18% had hepatic decompensation, 3% developed hepatocellular carcinoma, and 8% cleared HBsAg. Liver-related death was observed in 13% of patients and mainly occurred in patients from the first period (P = 0.012). These results indicate an outbreak of HDV at the end of the 1980s and the beginning of the 1990s, with a large number of acute HDV cases affecting predominately young, male intravenous drug users. Currently, patients with chronic HDV disease are older, and factors associated with worse prognosis include the presence of cirrhosis and age at the time of diagnosis.

Hepatitis delta virus (HDV) infection is associated with accelerated progression of fibrosis, early occurrence of hepatic decompensation and an increased risk for hepatocellular carcinoma. Epidemiological data on hepatitis delta virus (HDV) in Romania are still lacking. To assess the prevalence, virological, clinical and epidemiological features of HDV infection in Romanian patients. We conducted a multicenter study in 10 centers. Data on sociodemographic characteristics and potential risk factors were collected through a questionnaire. Virological markers of HBV and HDV infection, biochemical and clinical features of liver disease were evaluated. The study population comprised 2,761 HBsAg(+) patients with a mean age of 43.8+/-13.8 years, out of whom 5.2% were HBeAg(+) and 55.7% were males. Liver cirrhosis was detected in 17.9% of patients, while 80.4% had chronic hepatitis. The prevalence of IgG anti-HDV(+) patients was 23.1%, out of whom 16.4% were HDV RNA positive. The highest pr...

Hepatitis delta virus (HDV) superinfection of woodchuck chronic carriers of woodchuck hepatitis virus (WHV) results in acute and chronic disease. The different courses of disease mimicked the outcome of human HDV superinfection, making woodchucks valuable models for clinical studies of HDV. Ten of 11 woodchuck chronic carriers of WHV superinfected with HDV developed acute HDV infection with markers of viral replication in the serum and liver. One animal (DW128) had no serological markers of acute HDV infection. Nine of 11 (82%) superinfected animals developed chronic HDV infection. An unusual course of chronic HDV infection occurred in one woodchuck (DW128): no serum markers of acute or chronic HDV infection appeared but HDV RNA was detected in the liver, indicating that chronic HDV infection can occur without serological markers.

In vitro transcripts of bacteriophage RNA polymerases (RNAPs), such as T7 RNAP, often suffer from a considerable degree of 3′-end heterogeneity and, with certain promoter sequences, 5′-end heterogeneity. For some applications, this transcript heterogeneity poses a significant problem. A potential solution is to incorporate ribozymes into the transcripts at the 5′- and/or 3′-end of the target RNA sequence. This approach has been used quite widely but has required the generation of new transcription vectors or PCR-derived templates for each new RNA to be studied. To overcome this limitation, we have created two general plasmids for producing homogeneous RNA transcripts: one encodes a 3′- hepatitis delta virus (HDV) ribozyme and the other, used in combination with a two-step PCR, allows the production of double [5′-hammerhead (HH) and 3′-HDV] ribozyme constructs. A choice of cloning and run-off transcription linearisation restriction enzyme sites ensures that virtually any RNA sequence can be cloned and transcribed from these plasmids. For all the RNA sequences tested, good yields of transcript were obtained. These plasmids provide the tools for the simple, rapid creation of new RNA-coding plasmids to produce milligram quantities of homogeneous in vitro transcripts for all applications.

Hepatitis B and D viruses (HBV and HDV) are human pathogens with restricted host ranges and high selectivity for hepatocytes; the HBV L-envelope protein interacts specifically with a receptor on these cells. We aimed to identify this receptor and analyze whether it is the recently described sodium-taurocholate co-transporter polypeptide (NTCP), encoded by the SLC10A1 gene. To identify receptor candidates, we compared gene expression patterns between differentiated HepaRG cells, which express the receptor, and naïve cells, which do not. Receptor candidates were evaluated by small hairpin RNA silencing in HepaRG cells; the ability of receptor expression to confer binding and infection were tested in transduced hepatoma cell lines. We used interspecies domain swapping to identify motifs for receptor-mediated host discrimination of HBV and HDV binding and infection. Bioinformatic analyses of comparative expression arrays confirmed that NTCP, which was previously identified through a biochemical approach is a bona fide receptor for HBV and HDV. NTCPs from rat, mouse, and human bound Myrcludex B, a peptide ligand derived from the HBV L-protein. Myrcludex B blocked NTCP transport of bile salts; small hairpin RNA-mediated knockdown of NTCP in HepaRG cells prevented their infection by HBV or HDV. Expression of human but not mouse NTCP in HepG2 and HuH7 cells conferred a limited cell-type-related and virus-dependent susceptibility to infection; these limitations were overcome when cells were cultured with dimethyl sulfoxide. We identified 2 short-sequence motifs in human NTCP that were required for species-specific binding and infection by HBV and HDV. Human NTCP is a specific receptor for HBV and HDV. NTCP-expressing cell lines can be efficiently infected with these viruses, and might be used in basic research and high-throughput screening studies. Mapping of motifs in NTCPs have increased our understanding of the species specificities of HBV and HDV, and could lead to small animal models for studies of viral infection and replication.

Liver diseases linked to Hepatitis B-Hepatitis Delta virus co- or superinfections are more severe than during HBV mono-infection. Diagnosis of HDV infection therefore remains crucial in monitoring patients, but is often overlooked. To integrate delta markers into high throughput viral hepatitis diagnostic, we studied the binding of anti-delta antibodies (Abs) using surface plasmon resonance imaging (SPRi). We focused on the ubiquitous HDV genotype 1 (HDV1) and the more uncommon African-HDV6 and HDV8 genotypes to define an array with recombinant proteins or peptides. Full-length and truncated S-HDAg recombinant proteins of HDV1 and 11 HDV peptides of HDV1, 6 and 8, REPRESENTING VARIOUS PORTION OF THE DELTA ANTIGEN: were grafted onto biochips, allowing SPRi measurements. seventeen sera from patients infected with different HDV-genotypes were injected onto protein or peptide chips, RESPECTIVELY: . In all, Abs against HDV proteins and/or peptides were detected in 16 out of 17 infected p...