Honey Bee Dry Venom Reduces Hepatitis B Virus Surface Antigen Secretion in PLC/PRF/5 Cell Line

2019 ◽  
Author(s):  
Hussein Khani ◽  
Mahdi Ghorbani ◽  
Farshad Nojoomi ◽  
Alireza Mohebbi
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Line ◽  
Honey Bee ◽  
Surface Antigen ◽  
Post Treatment ◽  
Bee Venom ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
B Virus

Background and Aims: Currently, many efforts are directed toward functional Hepatitis B virus (HBV) treatment. This is achievable by suppression of HBV surface antigen (HBsAg) secretion. In this regard, use of natural products has been the areas of interest by scientific communities.  Materials and Methods: Dried Honey Bee venom was extracted for assessing its anti-HBsAg secretion potential. Hepatoma cell-derived PLC/PRF/5 was propagated in complete medium. The cell line was treated by a serial dilution of Bee venom. Cell cytotoxicity (IC50) was measured by MTT colorimetric assay at three post-treatment times. HBsAg secretion was evaluated from PLC/PRF/5 supernatant treated by under-cytotoxic concentrations of Bee venom by using enzyme-linked immunosorbent assay.  Results: The results indicated that dried Bee venom extract is able to reduce secretion of HBsAg from the cell line with Selectivity Index (SI) of eight. Reduced levels of HBsAg were in dose-dependent manner and it was in its lower concentrations at 8 ppm after 12 hr post treatment. The IC50 was observed to be 63.78 ppm. Conclusions: The Bee venom has anti-HBV activity. The way we used under-cytotoxic concentration of Bee venom, the HBsAg secretion was restored after 24 hr post treatment. Furthermore, mechanism of action of Bee venom in reducing HBsAg level needs to be further investigated.

scholarly journals Development and evaluation of an in-house ELISA to detect hepatitis B virus surface antigen in resource-limited settings

2014 ◽  
Vol 39 (2) ◽  
pp. 65-68 ◽  
Author(s):  
K Fatema ◽  
S Tabassum ◽  
A Nessa ◽  
M Jahan
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Hbv Infection ◽  
Health Concern ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Surface ◽  
B Virus ◽  
Commercial Kits ◽  
Checkerboard Titration

Hepatitis B virus (HBV) infection is of global public health concern. Among various serological tests used for the diagnosis and screening of HBV infection, the enzyme-linked immunosorbent assay (ELISA) to detect hepatitis B surface antigen (HbsAg) is most widely used. The present study was designed to develop and standardize a cost effective in-house ELISA for the detection of HbsAg and compare its performance with two established commercial kits. The concentrations of coating antibody, conjugates and sera were fixed by checkerboard titration. Using known HBsAg positive and negative sera, four different concentrations (1, 0.5, 0.25 and 0.125 ?g/well) of coating anti-HBs were applied. Similarly, serial dilutions of patients’ sera (1 in 2, 1 in 3, 1 in 5 and 1 in 9) and conjugates (1 in 2, 1 in 3, 1 in 5, 1 in 9 and 1 in 17) were evaluated by checkerboard titration. The optimal concentration of coating antibody was determined at 0.25 ?g/well and 1 in 9 dilution for both conjugates and sera. The performance comparison of our in-house ELISA showed excellent correlation with two commercial kits (Pearson 0.957, P=0.001 for monoclonal antibody coated kit and Pearson 0.929, P=0.000 for polyclonal antibody coated kit) when OD values were compared. All commercial kit proven positive samples was positive while all negative samples were negative with the in-house ELISA resulting in 100% sensitivity and specificity. The results of our study demonstrated that our inhouse ELISA for detection of HBsAg was equally as sensitive and specific as two well-known commercial kits. Thus, this system may be a useful tool for diagnostic and screening purposes, as well as outbreak investigations. DOI: http://dx.doi.org/10.3329/bmrcb.v39i2.19644 Bangladesh Med Res Counc Bull 2013; 39: 65-68

scholarly journals Expression Level of Small Envelope Protein in Addition to Sequence Divergence inside Its Major Hydrophilic Region Contributes to More Efficient Surface Antigen Secretion by Hepatitis B Virus Subgenotype D2 than Subgenotype A2

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 967
Author(s):  
Qianru Wang ◽  
Shuwen Fu ◽  
Jing Zhang ◽  
Quan Yuan ◽  
Jisu Li ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Envelope Protein ◽  
Hepatitis B Surface Antigen ◽  
Site Directed Mutagenesis ◽  
Enzyme Linked Immunosorbent Assay ◽  
S Protein ◽  
Huh7 Cells ◽  
B Virus

Hepatitis B surface antigen (HBsAg) promotes persistent hepatitis B virus (HBV) infection. It primarily corresponds to small (S) envelope protein secreted as subviral particles. We previously found that genotype D clones expressed less S protein than genotype A clones but showed higher extracellular/intracellular ratio of HBsAg suggesting more efficient secretion. The current study aimed to characterize the underlying mechanism(s) by comparing a subgenotype A2 clone (geno5.4) with a subgenotype D2 clone (geno1.2). Five types of full-length or subgenomic constructs were transfected to Huh7 cells at different dosage. HBsAg was quantified by enzyme linked immunosorbent assay while envelope proteins were detected by Western blot. We found that ratio of extracellular/intracellular HBsAg decreased at increasing amounts of DNA transfected. Conflicting findings from two types of subgenomic construct confirmed stronger secretion inhibitory effect of the genotype D-derived large envelope protein. Chimeric constructs followed by site-directed mutagenesis revealed geno1.2 specific V118/T127 and F161/A168 in the S protein as promoting and inhibitory of HBsAg secretion, respectively. In conclusion, more efficient HBsAg secretion by subgenotype D2 than subgenotype A2 is attributed to lower level of S protein expression in addition to V118 and T127 in S protein, although its F161 and A168 sequences rather reduce HBsAg secretion.

scholarly journals Development and Technical and Clinical Validation of a Quantitative Enzyme-Linked Immunosorbent Assay for the Detection of Human Antibodies to Hepatitis B Surface Antigen in Recipients of Recombinant Hepatitis B Virus Vaccine

2009 ◽  
Vol 16 (8) ◽  
pp. 1236-1246 ◽  
Author(s):  
Pierre Cambron ◽  
Jeanne-Marie Jacquet ◽  
Bernard Hoet ◽  
Marc Lievens
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Immunosorbent Assay ◽  
Surface Antigen ◽  
Hepatitis B Surface Antigen ◽  
Geometric Mean ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Limit Of Quantitation ◽  
B Virus

ABSTRACT Pending removal from the market of a commercial assay (the AUSAB [Abbott Laboratories] enzyme immunoassay [EIA]) for the determination of antibodies to hepatitis B surface antigen (HBsAg), a new in-house quantitative enzyme-linked immunosorbent assay (ELISA) to measure antibodies against HBsAg (anti-HBs) was developed (anti-HBs in-house). Specific anti-HBs antibodies were sandwiched between the precoated HBsAg ad and ay subtypes purified from plasma from hepatitis B virus (HBV) human carriers and the recombinant HBsAg adw2 subtype tagged with horseradish peroxidase. The assay was calibrated against the 1st International Reference Preparation for anti-hepatitis B immunoglobulin (lot 1977-W1042). Analytical sensitivity and the limit of quantitation were estimated at 0.43 mIU/ml and 2.0 mIU/ml, respectively. Overall reproducibility was 11.86%, and accuracy was estimated to be 94.89%. More than 4,000 samples from seven clinical trials were tested with the anti-HBs in-house assay and compared to results generated with AUSAB EIA and AUSAB radioimmunoassay (RIA). During the technical validation, the anti-HBs in-house assay was compared to the AUSAB RIA as a reference (n = 919). Overall assessment of concordance and Deming's regression analysis were performed. The coefficient of correlation between the AUSAB RIA and anti-HBs in-house assay was 0.9815 with a slope of 0.9187. The overall agreement between anti-HBs in-house and AUSAB RIA was 97.61%, considering the clinical cutoffs at 3.3 mIU/ml and 1.0 mIU/ml for the respective assays. From a clinical perspective, seroprotection rates and anti-HBs geometric mean antibody concentrations for individual studies calculated with either the in-house assay or the reference assays were similar. Conclusions of individual studies were confirmed. The performance characteristics of the in-house assay are acceptable. There is no evidence that use of the new assay would lead to different clinical conclusions from the reference method.

scholarly journals Sero-survey of Hepatitis B surface antigen amongst pregnant women attending Infectious Disease Hospital Bayara, Bauchi State, Nigeria

2012 ◽  
Vol 3 (1) ◽  
pp. 10
Author(s):  
James A. Ndako ◽  
Georgebest ON. Echeonwu ◽  
Obinna O. Nwankiti ◽  
Emamuzou M. Onovoh ◽  
Alloysius U. Jah ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Pregnant Women ◽  
Surface Antigen ◽  
Hepatitis B Surface Antigen ◽  
Neonatal Infection ◽  
Vaccination Schedule ◽  
Enzyme Linked Immunosorbent Assay ◽  
Public Health Importance ◽  
B Virus

Hepatitis B virus (HBV) continues to cause serious health problems in developing countries. Neonatal infection with HBV, which is often acquired during delivery, carries a high risk resulting in persistent infection. This research aims to detect the prevalence of Hepatitis B surface Antigen (HBsAg) among pregnant women in our location of study. One hundred and eighty (180) sera samples were screened among pregnant women aged 13-49, using standard enzyme-linked immunosorbent assay (ELISA) method. Structured questionnaire were administered to the subjects to obtain demographic and other relevant data. Overall result showed that 31 (17.2%) were found to be positive for HBsAg among the total subjects screened. The highest prevalence was found among those aged 20-29 with 11 (6.1%) seropositivity (x2=7.902; P=0.048). Considering occupational distribution of volunteer subjects, a high prevalence of 12 (6.7%); P<0.05 was recorded among house wives, which shows a measure of significance compared to other women screened. Furthermore, based on various risk factors subjects with history of surgery and use of unsterilized sharp instruments recorded 15 (8.3%) prevalence (P=0.233; P>0.05). How ever, women in their second trimester of pregnancy recorded a higher prevalence of 23 (12.8%):(P=0.080; P<0.05). This study therefore emphasizes the public health importance of HBV among pregnant women and equally suggests that children born to women with Hepatitis B Virus, be closely monitored for infection beyond the one and the half years of age, this also calls for a proper enlightenment on the dangers posed by the virus, while a well designed vaccination schedule is advocated among the general population.

Enhancement of hepatitis-B surface-antigen expression by 5-azacytidine in a hepatitis-B-virus-transfected cell line

1992 ◽  
Vol 52 (1) ◽  
pp. 137-140 ◽  
Author(s):  
Eiji Miyoshi ◽  
Junichi Fujii ◽  
Norio Hayashi ◽  
Keiji Ueda ◽  
Takahiro Towata ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Line ◽  
Surface Antigen ◽  
Hepatitis B Surface Antigen ◽  
Antigen Expression ◽  
Transfected Cell Line ◽  
B Virus ◽  
Surface Antigen Expression

Heat-regulated expression of the hepatitis B virus surface antigen in the human Wish cell line

1987 ◽  
Vol 8 (1) ◽  
pp. 43-59 ◽  
Author(s):  
Michel Dreano ◽  
Xavier Fouillet ◽  
Jean Brochot ◽  
Jean-Marie Vallet ◽  
Marie-Louise Michel ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Line ◽  
Surface Antigen ◽  
Virus Surface ◽  
Virus Surface Antigen ◽  
B Virus ◽  
Regulated Expression

scholarly journals Endocytosis of Hepatitis B Immune Globulin into Hepatocytes Inhibits the Secretion of Hepatitis B Virus Surface Antigen and Virions

2003 ◽  
Vol 77 (16) ◽  
pp. 8882-8892 ◽  
Author(s):  
Ralf Schilling ◽  
Samreen Ijaz ◽  
Michail Davidoff ◽  
Jia Yee Lee ◽  
Stephen Locarnini ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Immune Escape ◽  
Dependent Manner ◽  
Wild Type ◽  
Virus Surface ◽  
Virus Surface Antigen ◽  
B Virus ◽  
Specific Igg

ABSTRACT Hepatitis B immunoglobulin is used for prophylaxis against hepatitis B virus (HBV) and is thought to act by neutralization of virions and hepatitis B virus surface antigen (HBsAg)-containing particles in circulation. Using a panel of hepatocyte-derived cell lines, the present study investigated in vitro whether HBs-specific immunoglobulin G (IgG) is internalized in hepatocytes and whether it interacts with HBsAg in the cells. By immunoelectron microscopy and immunoblotting, human IgG and FcRn receptor for IgG were demonstrated on cellular membranes and in cytoplasmic extracts, irrespective of the HBsAg status of the cells. Furthermore, HBsAg and anti-HBs were shown to be colocalized in the same cellular compartment by two-color confocal microscopy. Endocytosis of HBs-specific IgG caused intracellular accumulation of HBsAg in a dose-dependent manner and inhibited the secretion of HBsAg and HBV virions from the cells. These effects were not observed with F(ab)2 fragments or nonimmune IgG as controls. The specificity of intracellular HBsAg- anti-HBs interaction was further investigated in cells transfected with HBV genomes expressing wild-type HBsAg or immune escape HBsAg (with a G145R mutation). Monoclonal anti-HBs markedly reduced the secretion of wild-type HBsAg, while the secretion of mutant HBsAg was not affected. These results suggest that HBs-specific IgG binds to hepatocytes and interacts with HBsAg within the cells. This may be relevant for the selection of surface antibody escape mutations.

Polypeptides of hepatitis B virus surface antigen produced by a hepatoma cell line.

1979 ◽  
Vol 32 (3) ◽  
pp. 796-802 ◽  
Author(s):  
P L Marion ◽  
F H Salazar ◽  
J J Alexander ◽  
W S Robinson
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Line ◽  
Surface Antigen ◽  
Hepatoma Cell ◽  
Hepatoma Cell Line ◽  
Virus Surface ◽  
Virus Surface Antigen ◽  
B Virus

scholarly journals Havachoobe (Onosma dichroanthum Boiss) Root Extract Decreases the Hepatitis B Virus Surface Antigen Secretion in the PLC/PRF/5 Cell Line

Intervirology ◽  
2020 ◽  
pp. 1-5
Author(s):  
Alireza Mohebbi ◽  
Fahimeh Azadi ◽  
Mohammad Mostakhdem Hashemi ◽  
Fatemeh Sana Askari ◽  
Nazanin Razzaghi
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Line ◽  
Real Time ◽  
Real Time Pcr ◽  
Time Dependent ◽  
Root Extract ◽  
Dependent Manner ◽  
B Virus ◽  
Dose Dependent

<b><i>Background:</i></b> Many efforts are currently focused on functional treatment of the hepatitis B virus (HBV). This can be done by suppressing the secretion of HBV surface antigen (HBsAg). Scientific communities are very interested in natural products in that respect. <b><i>Objective:</i></b> Use of root extract of Havachoobe (<i>Onosma dichroanthum BoissI</i>), a Northern Iranian native medical herb, for assessment of its anti-HBsAg secretion activity. <b><i>Methods:</i></b> Havachoobe had been bought at a nearby apothecary store. Plant root extract was obtained using a hydroalcoholic process. Cytotoxic activity of the extract was examined on PLC/PRF/5 cells using MTT assay. ELISA has been used to measure HBsAg in the treated cell line supernatants. In addition, real-time PCR analysis was performed to evaluate the expression of HBsAg before and after treatment of Onosma in vitro. <b><i>Results:</i></b> The results showed very low root extract cytotoxicity at concentrations under 8 μg/mL. Tissue culture infectious dose 50 was obtained at 63.78 μg/mL. In a dose-dependent and time-dependent manner, a significantly reduced HBsAg secretion was observed at a concentration of 8 ppm at 12 h post-treatment. The real-time PCR result showed relative decreased HBsAg expression at all doses at 12 h post-treatment time. <b><i>Discussion:</i></b> In this study, we first reported anti-HBsAg activity on an Iranian herbal medicine. Havachoobe root extract was shown to be able to inhibit HBsAg in a dose-dependent and time-dependent manner. We find the extract exerts its inhibitory effect of HBsAg by targeting transcription of HBsAg.

scholarly journals Agreement Between Rapid Hepatitis B Surface Antigen Test Kit and ELISA for the Diagnosis of Hepatitis B Virus Infection in Gondar, North West Ethiopia.

2021 ◽  
Author(s):  
Mulualem Lemma ◽  
Gueshay Tsegay ◽  
Kasaw Adane
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Hepatitis B Surface Antigen ◽  
Diagnostic Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Kit ◽  
B Virus ◽  
Kappa Value ◽  
Good Agreement

Abstract Background: Hepatitis B Virus is one of the major causes of global public health problems. Diagnosis of HBV is done by using ELISA and immune-chromatographic assays for detecting different serologic markers. Hepatitis B surface antigen (HBsAg) is the most frequently used serological marker employed to diagnosis HBV using Rapid Diagnostic Test (RDT) and Enzyme Linked Immunosorbent Assay (ELISA) methods. The RDT method is the most commonly used diagnostic tool in Ethiopia as well as in most African countries. Therefore assessing and knowing the diagnostic performance of the RDT is important. Method: A prospective cross sectional study was done at University of Gondar comprehensive specialized Hospital from Feb, 2017- May, 2017. Ethical clearance was obtained from School of Biomedical and Laboratory Sciences Research Ethics Committee. Permission was obtained from Blood Bank office to use blood sample collected from voluntary blood donors. HBsAg was diagnosed using RDT and ELISA methods. ELISA considered as gold-standard to the performance of RDT. Data were entered into statistical package for social sciences (SPSS) version 20 software and analysis was done by using SPSS and for sensitivity, specificity, predictive value, and likelihood ratio Medi calc software. The agreement of the methods presented assessed using kappa value. Result: Each 161 specimen was diagnosed using both ELISA (ADVANCED®) and RDT (Ecotest®). A total ELISA confirmed 41 positive specimens and 120 ELISA confirmed negative specimens were re-analyzed using RDT. The results of RDT showed only 35 positives from the total tested specimens including 29 which already positive by ELISA and RDT showing a sensitivity of 70.7% and specificity 95% with its kappa value 0.69 indicating a good agreement with ELISA.Conclusion and recommendation: Rapid Ecotest® test of HBsAg has good agreement with ELISA but low sensitivity and high false negative value, so there should be an algorithm for HBV diagnostic methods when we used a rapid test kits.

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