Investigating the hepatitis B virus life cycle using engineered reporter hepatitis B viruses

Nishitsuji, Hironori; Harada, Keisuke; Ujino, Saneyuki; Zhang, Jing; Kohara, Michinori; Sugiyama, Masaya; Mizokami, Masashi; Shimotohno, Kunitada

doi:10.1111/cas.13440

Cited by 17 publications

(11 citation statements)

References 23 publications

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“…170 The resulting HBV/NLuc particles entered target HepG2-NTCP cells and primary human hepatocytes in an NTCP-dependent manner and produced NLuc for at least 12 days after infection. 171 NLuc quantification theoretically evaluates the virus life cycle from the entry and cccDNA formation through core promoter-mediated transcription. In this system KX2–391 was identified as an inhibitor of the transcription mediated by a HBV precore promoter.…”

Section: Reporter Systems For Evaluating the Hbv Infection Processmentioning

confidence: 99%

Cell and Animal Models for Studying Hepatitis B Virus Infection and Drug Development

Lin

Chen

et al. 2019

Gastroenterology

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Many cell culture and animal models have been used to study hepatitis B virus (HBV) replication and its effects in the liver; these have facilitated development of strategies to control and clear chronic HBV infection. We discuss the advantages and limitations of systems for studying HBV and developing antiviral agents, along with recent advances. New and improved model systems are needed. Cell culture systems should be convenient, support efficient HBV infection, and reproduce responses of hepatocytes in the human body. We also need animals that are fully permissive to HBV infection, convenient for study, and recapitulate human immune responses to HBV and effects in the liver. High-throughput screening technologies could facilitate drug development based on findings from cell and animal models.

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Section: Reporter Systems For Evaluating the Hbv Infection Processmentioning

confidence: 99%

Cell and Animal Models for Studying Hepatitis B Virus Infection and Drug Development

Lin

Chen

et al. 2019

Gastroenterology

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“…After heat inactivation, a slight decrease in the viral load was observed ( Table 2 ), as described by others [ 59 ]. Probably, viable particles were not present in those samples, since plasma samples with no more than 4 log 10 copies/mL were selected for this study, and HepG2 cell lines were not susceptible to HBV or HAV replication [ 60 – 62 ].…”

Section: Discussionmentioning

confidence: 99%

Complement System as a Target for Therapies to Control Liver Regeneration/Damage in Acute Liver Failure Induced by Viral Hepatitis

Melgaço

Veloso

Pacheco-Moreira

et al. 2018

Journal of Immunology Research

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The complement system plays an important role in innate immunity inducing liver diseases as well as signaling immune cell activation in local inflammation regulating immunomodulatory effects such as liver damage and/or liver regeneration. Our aim is to evaluate the role of complement components in acute liver failure (ALF) caused by viral hepatitis, involving virus-induced ALF in human subjects using peripheral blood, samples of liver tissues, and ex vivo assays. Our findings displayed low levels of C3a in plasma samples with high frequency of C3a, C5a, and C5b/9 deposition in liver parenchyma. Meanwhile, laboratory assays using HepG2 (hepatocyte cell line) showed susceptibility to plasma samples from ALF patients impairing in vitro cell proliferation and an increase in apoptotic events submitting plasma samples to heat inactivation. In summary, our data suggest that the complement system may be involved in liver dysfunction in viral-induced acute liver failure cases using ex vivo assays. In extension to our findings, we provide insights into future studies using animal models for viral-induced ALF, as well as other associated soluble components, which need further investigation.

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“…This could be a better system for many purposes but essentially this approach cannot be utilized by the researchers when they need to study the viral life cycle and influence of variants of various kinds on the viral life cycle. 36,37 The approaches demonstrated in our present study could be a better approach.…”

mentioning

confidence: 84%

Impact of length of replication competent genome of hepatitis B virus over the differential antigenic secretion

Amir

Siddiqui

Farooqui

et al. 2019

J of Cellular Biochemistry

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Hepatitis B virus (HBV) genome consists of circular partially double stranded DNA of 3.2 kb size which gets converted into covalently closed circular DNA (cccDNA) during its life cycle. It then acts as a template for formation of pregenomicRNA (pgRNA) of 3.5 kb. Absence of appropriate animal models prompted a need to establish a better in vitro culture system to uncover the propagation and survival mechanisms of the virus. There is scarcity of data to represent the significance of varying length of replication competent viral genome on the secretion of viral secretory proteins/antigens and in turn on the overall effects on the accomplishment of the viral life cycle. The present study was undertaken to ascertain a suitable replication competent construct in which the viral life cycle of HBV with varying clinical relevance can be studied efficiently. Two constructs (pHBV 1.3 and pHBV 1X) of different sizes were used to transfect hepatoma cells and consequently the secretory antigens were monitored. In vector free approach (pHBV 1X), 3.2 kb viral DNA is directly transfected in the culture system whereas in vector mediated approach more than full length of viral genome is cloned in a vector (pHBV 1.3X) and transfected to obtain a 3.5 kb pgRNA intermediate. HBV secretes two important antigens; HBsAg and HBeAg. HBsAg is a hallmark of infection and is the first to be secreted in the blood stream whereas HBeAg is a secretory protein and remains associated with the viral replication. The construct pHBV 1.3X referring to as more than full length, by virtue of being capable of undergoing transcription without the synthesis of cccDNA intermediate (unlike the clinical situation where an intermediate step of cccDNA synthesis is an essential component to initiate the viral life cycle) appears to be better system for studying viral life cycle in in vitro culture system. The reasons could be assigned to the fact that as low as 100 ng of viral DNA was shown to quantify the replicative phenotypes with this construct. The better efficiency of this construct at prima facie, appears to be mediated through the significantly higher levels of pgRNA transcript during the viral life cycle.

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Investigating the hepatitis B virus life cycle using engineered reporter hepatitis B viruses

Cited by 17 publications

References 23 publications

Cell and Animal Models for Studying Hepatitis B Virus Infection and Drug Development

Cell and Animal Models for Studying Hepatitis B Virus Infection and Drug Development

Complement System as a Target for Therapies to Control Liver Regeneration/Damage in Acute Liver Failure Induced by Viral Hepatitis

Impact of length of replication competent genome of hepatitis B virus over the differential antigenic secretion

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