Characterization of Novel Hepatitis B Virus PreS/S-Gene Mutations in a Patient with Occult Hepatitis B Virus Infection

Chen, Jianhong; Liu, Yan; Zhao, Jingbo; Xu, Zhihui; Chen, Rongjuan; Liu, Si; Lu, Shanshan; Li, Xiaodong; Wang, Shuai; Zhang, Kai; Li, Jin; Han, Junfeng; Xu, Da

doi:10.1371/journal.pone.0155654

Cited by 45 publications

(59 citation statements)

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“…Reduced expression levels of HBsAg because of mutations in the surface protein coding region have been reported previously [20,21]. Accordingly, all three sets of mutations resulted in a significant decrease in surface protein levels (Fig.…”

Section: Resultssupporting

confidence: 74%

Selection of the highly replicative and partially multidrug resistant rtS78T HBV polymerase mutation during TDF-ETV combination therapy

Shirvani-Dastgerdi

Winer

Celià-Terrassa

et al. 2017

Journal of Hepatology

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Background & Aims Patients chronically infected with the hepatitis B virus (HBV) that are on long-term treatment with nucleoside or nucleotide analogues are at risk of selecting HBV strains with complex mutational patterns. We herein report two cases of HBV-infected patients with insufficient viral suppression despite dual antiviral therapy with entecavir (ETV) and tenofovir (TDF), of which one patient died from aggressive hepatocellular carcinoma (HCC). Methods Serum samples of the patients from different time-points were analyzed by ultra-deep pyrosequencing analysis. Identified HBV mutations were functionally analyzed after transient transfection of replication-competent HBV vectors into hepatoma cells in vitro. We assessed replication efficacy, resistance to antivirals and potential impact on HBV secretion (viral particles, exosomes). Results Sequencing analyses revealed the selection of the rtS78T HBV polymerase mutation in both cases that simultaneously creates a premature stop codon at sC69 and thereby deletes almost the entire small HBV surface protein. One of the patients had an additional 261-bp deletion in the preS1/S2 region. Functional analyses of the mutations in vitro revealed that the rtS78T/sC69* mutation, but not the preS1/S2 deletion, significantly enhanced viral replication and conferred reduced susceptibility to ETV and TDF. The sC69* mutation caused truncation of HBs protein, leading to impaired detection by commercial HBsAg assay, without causing intracellular HBsAg retention or affecting HBV secretion. Conclusions The rtS78T/c69* HBV mutation associated with enhanced replication and insufficient response to antiviral treatment may favor long-term persistence of these isolates. Along with increased production of HBV transcripts and the sustained secretion of viral particles in the absence of antigenic domains of S protein, this HBV mutation may predispose to carcinogenic effects.

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Section: Resultssupporting

confidence: 74%

Selection of the highly replicative and partially multidrug resistant rtS78T HBV polymerase mutation during TDF-ETV combination therapy

Shirvani-Dastgerdi

Winer

Celià-Terrassa

et al. 2017

Journal of Hepatology

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“…We recently identified 60 OBI blood donors in Baoji area of North China and verified that a few mutations in the MHR of HBV S gene were more frequently detected in OBI blood donors compared to controls . In addition, multiple OBI‐related mutational patterns were identified from an OBI patient and five of them in the MHR were not documented previously . However, how these mutations contribute to undetectable HBsAg remains the need for clarification.…”

Section: Introductionmentioning

confidence: 80%

“…Noticeably, all S‐gene mutants had lower but not completely absent HBsAg, which is not fully consistent with OBI presentation of studied persons. This could be explained by existence of low‐level of anti‐HBs in their serum samples, because low‐affinity or non‐neutralizing antibodies induced by mutant HBsAg may play a role in OBI by interfering with HBsAg detection. In fact, detection of the OBI‐mutations may not definitely cause OBI presentation in clinic.…”

Section: Discussionmentioning

confidence: 99%

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Antigenicity reduction contributes mostly to poor detectability of HBsAg by hepatitis B virus (HBV) S‐gene mutants isolated from individuals with occult HBV infection

Zhang

Liu

Chen

et al. 2017

Journal of Medical Virology

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Mutations in hepatitis B virus (HBV) S gene are one of factors contributing to occult HBV infection (OBI). The study aimed to uncover the impact of OBI-related S-gene mutations on the detectability of hepatitis B surface antigen (HBsAg). Nine representative mutations within the major hydrophilic region of the S region were investigated. These included six (M1-M6) from an OBI patient with HBV-related hepatocellular carcinoma, and three (M7-M9) from three OBI blood donors. Recombinant plasmids on the basis of pTriEx-mod-1.1 HBV and pcDNA3.1(-)/myc-His A vectors were constructed for each and transfected into HepG2 or Huh7 cells, respectively. Electrochemical luminescence, ELISA, Western blotting, and confocal immunofluorescence were used to examine HBsAg expression and antigenicity. In comparison to wild-type strain, supernatant and intracellular HBsAg levels of the nine mutants were reduced by 56.39-99.09% and 42.76-99.77% upon Roche quantitative Elecsys assay, respectively. Confocal immunofluorescence showed that relative intensity ratios of HBsAg-myc-His fusion protein detected by anti-HBs and anti-His-tag were lower by 11.87-76.27% for the nine mutants compared to the wild-type strain. Specifically, M1-M5 mutants that we firstly found recently were 33.14%, 76.27%, 57.93%, 53.37%, and 40.88% lower, respectively. Consistent results were obtained using double-antibody sandwich ELISA assays (anti-myc + anti-HBs vs anti-myc + anti-His). Antigenicity reduction played a major role for the poor detectability of HBsAg caused by the OBI-related mutations, although decreased HBsAg expression of some mutants and anti-HBs in samples might play coordinated roles. Taken together, antigenicity reduction contributes mostly to poor detectability of HBsAg caused by these OBI-related mutations.

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“…Finally, the cryptic pathogenesis of OBI has been a major focus of research over the past three decades, with the most compelling viral molecular mechanism underlying the infection suggested to be mutations occurring within the viral HBsAg‐encoding gene and the overlapping polymerase (pol) gene frame . These OBI‐associated variations have been extensively investigated . However, a drawback to the majority of previous investigations has been the cross‐sectional study designs, resulting in a knowledge gap on the genetic stability of the variations over time.…”

Section: Introductionmentioning

confidence: 99%

Evolution of the serologic and virologic course of occult HBV infection in therapy experienced HIV co‐infected patients

Amponsah-Dacosta

Selabe

Mphahlele

2017

Journal of Medical Virology

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We investigated how the natural course of occult hepatitis B virus (HBV) infection (OBI) may evolve during HIV co-infection and long term HBV-active HAART. From a cohort of 181 HIV infected patients who were consecutively recruited over a 5 year period, 28 HBV co-infected patients with sequential sera (n = 98) were identified. Iterative HBV serology and viral loads were determined before and during treatment. The viral HBsAg gene was then serially amplified, directly sequenced, and molecularly characterized. Persistent detection of anti-HBs did not result in a modification to the clinical course of OBI. In contrast, reactivation of chronic HBV infection, hepatic enzymatic flares and cases of HBV reinfection were evident among anti-HBs negative OBI patients, and this was a notable finding. Of the 14 chronic HBV infected patients, eight progressed to persistent OBI after initiation of HBV-active HAART, increasing the number of patients with OBI in the study. Long term HBV-active HAART was not found to have a notable impact on low level viremia during OBI. While the HBsAg gene sequences isolated from chronic HBV infection were genetically stable over time, OBI-associated variants (sP111R, sT127P, sY161F) were neither stable nor predominant during the course of infection. This study is the first of its kind from South Africa to show the occurrence of hepatic enzymatic flares, HBV reactivation, and reinfection in HAART-exposed HIV co-infected patients with OBI. Among the cases studied, there was further evidence that OBI-associated variants may not play a significant role in the pathogenesis of OBI.

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Characterization of Novel Hepatitis B Virus PreS/S-Gene Mutations in a Patient with Occult Hepatitis B Virus Infection

Cited by 45 publications

References 54 publications

Selection of the highly replicative and partially multidrug resistant rtS78T HBV polymerase mutation during TDF-ETV combination therapy

Selection of the highly replicative and partially multidrug resistant rtS78T HBV polymerase mutation during TDF-ETV combination therapy

Antigenicity reduction contributes mostly to poor detectability of HBsAg by hepatitis B virus (HBV) S‐gene mutants isolated from individuals with occult HBV infection

Evolution of the serologic and virologic course of occult HBV infection in therapy experienced HIV co‐infected patients

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