The composition of the hepatitis B surface antigen (HBsAg) in acute hepatitis B virus (HBV) infections is reproducible in an in vitro model

Patients undergoing chronic hemodialysis, as well as dialysis staff members, are at high risk of infection with hepatitis B virus (HBV). We have analyzed by PCR the presence of HBV DNA in serum and peripheral blood mononuclear cells (PBMC) from 33 hepatitis B surface antigen (HBsAg)-negative hemodialysis patients and 24 dialysis unit staff members; eight of the 24 staff members had an acute hepatitis B resolved 13 to 21 yr before. HBV DNA was detected in serum of 19 (58%) patients (12 of 17 with and 7 of 16 without anti-HBV antibodies). HBV DNA was found in PBMC of 18 (54%) patients (13 of 17 with and 5 of 16 without anti-HBV antibodies). In the staff members, serum HBV DNA was found only in the individuals who suffered a previous acute hepatitis (P < 0.005). HBV DNA was detected in PBMC of four of six staff members (all with previous acute hepatitis). In two HBV DNA-positive PBMC samples, viral RNA was detected by reverse transcription-PCR. To ascertain whether the HBV DNA detected in serum was encapsulated, seven HBV DNA-positive serum samples were digested with DNase before PCR. After treatment, HBV DNA remained detectable in four cases. In conclusion, HBV DNA in serum and PBMC is detectable in a high proportion of HBsAg-negative hemodialysis patients and may persist several years after a resolved acute hepatitis B. The viral DNA is encapsulated and remains transcriptionally active in PBMC. In the anti-HBs-negative patients, HBV DNA is, at the present time, the only means for diagnosing a past HBV hepatitis.

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Primary Tupaia Javanica Hepatocyte Culture as an In Vitro Model for Human Hepatitis B Virus Infection

The Indonesian Journal of Gastroenterology Hepatology and Digestive Endoscopy ◽

10.24871/2232021203-209 ◽

2022 ◽

Vol 22 (3) ◽

pp. 203-209

Author(s):

Kemal Fariz Kalista ◽

Maryati Surya ◽

Silmi Mariya ◽

Diah Iskandriati ◽

Irsan Hasan ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

In Vitro Model ◽

Hbv Dna ◽

Hbv Infection ◽

Primate Research ◽

Qualitative And Quantitative ◽

B Virus ◽

Vitro Model

Background: Hepatitis B virus (HBV) infection is still one of the biggest health problems in the world, which could lead to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Treatment for HBV infection has not yet achieved a functional cure. More studies are needed to investigate human HBV (HuHBV), but the scarcity of animal models for HuHBV infection became a barrier. Recently, many studies have shown that Tupaia are suitable for the study of HuHBV. The purpose of this study was to develop a primary tupaia hepatocyte (PTH) culture from T. javanica, a species of Tupaia found in Indonesia, and to prove that HuHBV can replicate in the PTH.Method: In vitro experimental study using PTH isolated from five wild adult T. javanica in Primate Research Center, IPB University. HuHBV was taken from humans with HBsAg and HBV-DNA (+). PTH cells then were infected with HuHBV after reaching 80% confluence. Observation on PTH cells was done everyday for 20 days. Qualitative and quantitative HBsAg were measured using a CMIA while HBV-DNA and cccDNA were measured by RT-PCR.Results: A cytopathic effect was seen on day post infection (DPI)-16. HBsAg and HBV-DNA were detected from DPI-2 until DPI-18, with HBV-DNA level peaked on DPI-12. cccDNA concentration was fluctuating from DPI-2 until DPI-20 with highest level on DPI-16.Conclusion: HuHBV could infect and replicate in PTH from T. javanica can be infected with HuHBV and HuHBV can replicate in the PTH from T. javanica.

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In vitro synthesis of antibody to hepatitis B virus antigen by circulating lymphocytes from chronic HBsAg carriers and patients with acute hepatitis B

Gastroenterologia Japonica ◽

10.1007/bf02774832 ◽

1986 ◽

Vol 21 (2) ◽

pp. 145-151 ◽

Cited By ~ 1

Author(s):

Kenichi Murase ◽

Shinichi Kakumu ◽

Akinori Tsubouchi ◽

Kentaro Yoshioka ◽

Nobuo Sakamoto

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Acute Hepatitis ◽

Virus Antigen ◽

In Vitro Synthesis ◽

Hbsag Carriers ◽

B Virus ◽

Acute Hepatitis B ◽

Circulating Lymphocytes

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In Vitro Systems for Studying Different Genotypes/Sub-Genotypes of Hepatitis B Virus: Strengths and Limitations

Viruses ◽

10.3390/v12030353 ◽

2020 ◽

Vol 12 (3) ◽

pp. 353 ◽

Cited By ~ 1

Author(s):

Constance N. Wose Kinge ◽

Nimisha H. Bhoola ◽

Anna Kramvis

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

In Vitro Model ◽

Antiviral Agents ◽

Model Systems ◽

Effective Vaccine ◽

B Virus ◽

In Vitro Systems ◽

Vitro Model

Hepatitis B virus (HBV) infects the liver resulting in end stage liver disease, cirrhosis, and hepatocellular carcinoma. Despite an effective vaccine, HBV poses a serious health problem globally, accounting for 257 million chronic carriers. Unique features of HBV, including its narrow virus–host range and its hepatocyte tropism, have led to major challenges in the development of suitable in vivo and in vitro model systems to recapitulate the HBV replication cycle and to test various antiviral strategies. Moreover, HBV is classified into at least nine genotypes and 35 sub-genotypes with distinct geographical distributions and prevalence, which have different natural histories of infection, clinical manifestation, and response to current antiviral agents. Here, we review various in vitro systems used to study the molecular biology of the different (sub)genotypes of HBV and their response to antiviral agents, and we discuss their strengths and limitations. Despite the advances made, no system is ideal for pan-genotypic HBV research or drug development and therefore further improvement is required. It is necessary to establish a centralized repository of HBV-related generated materials, which are readily accessible to HBV researchers, with international collaboration toward advancement and development of in vitro model systems for testing new HBV antivirals to ensure their pan-genotypic and/or customized activity.

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Specific expression of human interferon-gamma controls hepatitis B virus replication in vitro in secreting hepatitis B surface antigen hepatocytes

Journal of Virological Methods ◽

10.1016/j.jviromet.2011.12.016 ◽

2012 ◽

Vol 180 (1-2) ◽

pp. 84-90 ◽

Cited By ~ 3

Author(s):

Quancheng Kan ◽

Duolu Li ◽

Zujiang Yu

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Interferon Gamma ◽

Virus Replication ◽

Surface Antigen ◽

Hepatitis B Surface Antigen ◽

Human Interferon ◽

Specific Expression ◽

B Virus

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Improved Detection of Hepatitis B Virus Surface Antigen by a New Rapid Automated Assay

Journal of Clinical Microbiology ◽

10.1128/jcm.37.8.2639-2647.1999 ◽

1999 ◽

Vol 37 (8) ◽

pp. 2639-2647 ◽

Cited By ~ 35

Author(s):

Bernard Weber ◽

Anja Bayer ◽

Peter Kirch ◽

Volker Schlüter ◽

Dietmar Schlieper ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Acute Hepatitis ◽

Sensitivity And Specificity ◽

Surface Antigen ◽

Core Antigen ◽

B Virus ◽

Acute Hepatitis B ◽

Screening Assays ◽

Hbsag Screening

The performance of hepatitis B virus (HBV) surface antigen (HBsAg) screening assays is continuously improved in order to reduce the residual risk of transfusion-associated hepatitis B. In a multicenter study, a new automated rapid screening assay, Elecsys HBsAg (Roche Diagnostics), was compared to well-established tests (Auszyme Monoclonal [overnight incubation] version B and IMx HBsAg [Abbott]). Included in the evaluation were 23 seroconversion panels; sera from the acute and chronic phases of infection; dilution series of various HBsAg standards, HBV subtypes, and S gene mutants; and isolated anti-HBV core antigen-positive samples. To challenge the specificity of the new assay, sera from HBsAg-negative blood donors, pregnant women, and dialysis and hospitalized patients and potentially cross-reactive samples were investigated. Elecsys HBsAg showed a higher sensitivity for HBsAg subtypes ad, ay, adw2, adw4, ayw1, ayw2, ayw4, and adr detection in dilution series of different standards or sera than Auszyme Monoclonal version B and/or IMx HBsAg. Acute hepatitis B was detected in 11 to 16 of 23 seroconversion panels between 2 and 16 days earlier with Elecsys HBsAg than with the alternative assays. Elecsys HBsAg and Auszyme Monoclonal version B detected HBsAg surface mutants with equal sensitivity. The sensitivity and specificity of Elecsys HBsAg were 100%. Auszyme Monoclonal version B had a 99.9% specificity, and its sensitivity was 96.6%. IMx HBsAg showed a poorer sensitivity and specificity than the other assays. In conclusion, Elecsys HBsAg permits earlier detection of acute hepatitis B and different HBV subtypes than the alternative assays. By using highly sensitive HBsAg screening assays, low-level HBsAg carriers among isolated anti-HBV core antigen-positive individuals can be detected.

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