Replicative and Transcriptional Activities of Hepatitis B Virus in Patients Coinfected with Hepatitis B and Hepatitis Delta Viruses (original) (raw)
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Journal of Hepatology, 2010
Background & Aims: This study presents a real-time reversetranscription PCR (rt-RT-PCR) assay for hepatitis delta virus (HDV) RNA quantification, designed to clarify the interplay between HDV and hepatitis B virus (HBV) in chronic infection. Methods: Serum HDV-RNA and HBV-DNA were analysed by rt-RT-PCR in a cross-sectional study of 37 untreated chronic HDV patients, 25 of whom were also longitudinally studied. Results: In the cross-sectional study, both viruses were active in 15 (40.5%) patients and inactive in 4 (10.8%); HDV alone was active in 12 (32.4%) and HBV in 6 (16.2%). The longitudinal study showed seven replication profiles, with considerable fluctuating activity of one or both viruses, including alternating predominance. In 20% of cases, longitudinal HBV/HDV viral loads differed from cross-sectional results, indicating a risk of misinterpreting HBV/HDV interactions when assessing a single determination. Fluctuating HBV replication only increased in the presence of fluctuating HDV activity. HBsAg levels, stable in HBV single infection, fluctuated in HDV chronic infection. The results of both the cross-sectional and longitudinal study call into question the major suppressor effect of HDV over HBV, revealing an important role of HBV. Conclusions: Longitudinal evaluation of viremia shows a complex interaction between HBV and HDV and is essential to understand the pathophysiology of chronic HDV infection. Ó consecutive Spanish Caucasian patients with chronic HBV/HDV infection admitted to our hospital were included in this study. All patients had histological evidence of chronic hepatitis, serum alanine amino-Journal of Hepatology 2010 vol. 52 j 658-664
Quantification of intrahepatic hepatitis B virus (HBV) DNA in patients with chronic HBV infection
Hepatology, 2000
No data are available about the amount of hepatitis B virus (HBV) genomes in liver of patients with chronic HBV infection. The aim of this study was to quantify the intrahepatic HBV DNA in hepatitis B surface antigen (HBsAg)-positive patients with either active or suppressed viral replication and in HBsAg-negative subjects with occult HBV infection. We optimized the Roche ''Amplicor HBV Monitor'' kit for quantifying liver HBV DNA and analyzed hepatic DNA extracts and serum samples from 19 HBs-Ag-positive and 43 HBsAg-negative individuals. Eight of the HBsAg carriers had active HBV replication, and for 3 of them we analyzed samples obtained before and at the end of 1 year of lamivudine treatment. Five hepatitis Delta virus (HDV) coinfected patients and 6 healthy HBsAg carriers had inhibited HBV activity. Among the HBsAg-negative subjects 21 had occult HBV infection and 22 had no evidence of HBV infection. The median of HBV genomes per microgram of liver DNA milliliter of serum was 34,500 to 2,620,000 in patients with active viral replication, 20,000 to 3,900,000 before and 10,000 to 2,800 at the end of therapy in lamivudine-treated individuals, 9,800 to 600 in HDVinfected individuals, and 7,450 to 17,400 in healthy HBsAg carriers. These data indicate that cases with suppressed HBV activity, despite the very low levels of viremia, maintain a relatively high amount of intrahepatic viral genomes. This virus reservoir is likely involved in HBV reactivation, which is usually observed after stopping lamivudine treatment. Finally, the analysis of cases with occult HBV infection showed that the assay we used was able to specifically detect and quantify as few as 100 copies of viral genomes per microgram of liver DNA. (HEPATOLOGY 2000;31: 507-512.)
The Journal of Infectious Diseases, 2021
Hepatitis B virus (HBV) DNA and RNA were quantified by digital PCR assays in 20–30 tissue pieces from each of 4 liver explants with cirrhosis caused by HBV. The within-patient variability of HBV RNA levels between pieces was up to a 1000-fold. Core RNA and S RNA levels were similar and correlated strongly when replication was high, supporting that transcription was from covalently closed circular DNA (cccDNA). By contrast, enhanced expression of S RNA relative to cccDNA and core RNA in patients with medium-high or low replication supports that HBV surface antigen (HBsAg) can be expressed mainly from integrated HBV DNA in such patients.
Detection of hepatitis B virus transcripts in patients with chronic liver disease
Journal of Hepatology, 1990
Notthem blot aoalyxis o" total cellular RNA pwitied from liver biopsies in 70 patients with chronic liver disease (24 HBsAg positive. 15 autiHE% andlor a"tiHRc positive, 31 HBV aeg-at&). No transcripts were found in the HBV negative and in the autiHBr and/or antiHBc positive patients. I" the others. three major RNA species were identified: i. a 3.5 kb transcript correspooding to the RNA pregenomc; ii. 2.4-2.1 kbtmoscript corresponding to the s ar.d preS1 gene RNA; iii. lower mokdar weight species. All three forms were pnscnt simulraneously only in patients with sctive viral replication, with a strict relation between the presz%ce of the 3.5 Lb RNA ia the liver and serum HBV-DNA. II. conclusion, Northern blot analysis ca" easily be performed to study viral replication and it can contribute to a better understanding of the molecular processes uoderiying HBV infcetion and leading to liver disease in ma".
Quantitative HBsAg and HDV-RNA levels in chronic delta hepatitis
Liver International, 2010
Background: Hepatitis delta virus (HDV) causes severe liver disease. Aims: To investigate the quantitative HDV-RNA, HBsAg and hepatitis B virus (HBV)DNA levels in correlation to histological, biochemical and demographical parameters in patients with chronic HDV infection as similar data in a large series of HDV patients are missing. Methods: Eighty HDV patients were recruited in Germany, Turkey and Greece; quantitative determination of HDV-RNA, HBsAg and HBV-DNA was performed by real-time polymerase chain reaction, the Architect HBsAg assay and Cobas TaqMan HBV test respectively. Results: All patients were infected with HDV-genotype 1. Thirtyfive patients (48%) had significant fibrosis (Ishak 3-4) and 15 (20.5%) had cirrhosis. HDV viraemia ranged from 1.
The Journal of general virology, 1998
Hepatitis B virus (HBV) isolates with A-1762 to T and G-1764 to A mutations in the core promoter have been associated with active hepatitis, severe liver disease following liver transplantation, hepatocellular carcinoma and acute fulminant courses--in the latter case combined with a C-1653 to T mutation. In this study, a mutant core promoter region containing the T-1653, T-1762 and A-1764 mutations was placed into the context of a wild-type HBV genome and analysed by transfection. The mutations reduced the level of pre-C mRNA (by 55%) and e-antigen secretion. In contrast, no significant effects on the levels of pregenome/C and pre-S/S mRNAs, intracellular core, polymerase, and pre-S /S2 proteins and secreted S-antigen were observed. The amount of progeny virus DNA in the cells and in the culture medium was increased marginally, if at all.
HBV DNA viral load and chronic hepatitis infection in different stages
Hepatitis Monthly, 2005
Although most studies addressing this question have shown a correlation between virus serum titer and severity of liver damage, some others have failed to show any correlation between them. The aim of the present study was to determine any correlation between HBV viral load and the severity of liver disease. Methods: 200 patients, including inactive carriers (n=104), patients with chronic active hepatitis (n=74: 55 HBeAg negative and 19 HBeAg positive) and with cirrhosis (n=22) entered the study. Quantitative serum HBV RNA assay was carried out using the PCR Amplicore technique.
Viruses, 2022
A comprehensive characterization of chronic HBV (CHB) patients is required to guide therapeutic decisions. The cumulative impact of classical and novel biomarkers on the clinical categorization of these patients has not been rigorously assessed. We determined plasma HBV-RNA and HBsAg levels, HBV in peripheral lymphocytes (PBMCs) and HBV mutation profiles in CHB patients. Patient demographics (n = 139) and classical HBV biomarkers were determined during a clinical routine. HBV-RNA in plasma and HBV-DNA in PBMCs were determined by RT-PCR. HBsAg levels were determined using Architect. In samples with HBV-DNA viral load >1000 IU/mL, genotype mutations in precore (PC), basal core promoter (BCP), HBsAg and Pol regions were determined by sequencing. Most patients (n= 126) were HBeAg-negative (HBeAgNeg) with significantly lower levels of HBV-RNA, HBV-DNA and HBsAg compared to HBeAg-positive (HBeAgPos) patients (p < 0.05). HBV genotype D prevailed (61/68), and >95% had BCP/PC mutati...
Journal of virology, 1997
Hepatitis B virus (HBV) has been reported to exist in peripheral blood mononuclear cells (PBMC), but it is not clear whether it replicates there. A precondition for replication should be the formation of covalently closed viral DNA and transcription of all essential viral mRNAs. The mRNAs of HBV form a nested box with common 3' ends. In order to detect even low levels of potential replication, we developed a quantitative reverse transcription-PCR method for detection of a smaller HBV mRNA species in the presence of the larger ones. All three highly viremic patients tested so far had mRNAs for the large and the small surface proteins and the X protein of the virus within PBMC but not in the virus from their sera. Furthermore, we detected by PCR covalently closed viral DNA in their PBMC. These data suggest that HBV may be not only taken up but also replicated by mononuclear blood cells and that these cells may be an extrahepatic site of viral persistence. X mRNA was detected in th...
The Southeast Asian journal of tropical medicine and public health, 2010
Covalently closed circular DNA (cccDNA) is a unique episomal replicative intermediate molecule of hepatitis B virus (HBV) which plays a key role in viral persistence. The aim of this study was to prove cccDNA persistence in the liver tissue of patients negative for hepatitis B surface antigen (HBsAg) and positive for antibody to hepatitis core antigen (anti-HBc). Intrahepatic HBV DNA and cccDNA were determined using real-time and semi-nested PCR assays on the liver tissues of 35 patients who were negative for HBsAg and positive for anti-HBc with or without anti-HBs. HBV DNA was detected in the liver tissue of 4 out of 35 patients who were positive for anti-HBc. None of the samples harbored cccDNA. In this study population, which is of Asian origin, very low levels of HBV DNA were detected in a small percentage of patients with anti-HBc. Even using the highly sensitive semi-nested PCR assay, HBV cccDNA was not detectable in any anti-HBc positive patients either with or without anti-HBs.