Purpose : Although there are a lot of the reports about the persistence of anti HBs titer of plasma derives HBV vaccine, it is difficult to find the follow up studies of the recombinant HBV vaccine. We performed this study to compare the persistence of anti HBs titer by vaccination schedule and the seronegative rate of 5 years later according to Anti HBs titer after basic immunization in neonatal period by recombinant HBV vaccination. Methods : This study was performed on 420 neonates at Pusan Moon Hwa Hospital from April to December 1993, followed up for 5 years after basic immunization by recombinant HBV vaccine. The anti HBs titer test was done by radioimmunoassay(RIAAUSAB, Abbott laboratories). The positive anti HBs level that would protect against HBV infection was defined as a level equal to or greater than 10mIU/mL. Results : In this study the seronegative rate after 5 years was 5% in 2 month schedule group, 25.5% in 6 month schedule group(P>0.05). In 2 month schedule group the seronegative rate was 20% when anti HBs titer is lower than 200mIU/mL, 0% when more than 200mIU/mL(P>0.05). In 6 month schedule group the seronegative rate was 66.7% when anti HBs titer was lower than 200mIU/mL, 40% when 200~499.9mIU/mL, 23.9% when 500~999.9mIU/mL, 22.5% when more than 1000mIU/mL. Conclusion : In this study the seronegative rate after 5 years of recombinant HBV vaccination was 5~25.5%. The persistence of anti HBs titer was statistically irrelevant to schedule. The seronegative rate after 5 years was statistically irrelevant to anti HBs titer after basic immunization.
Hu, Bo;Liang, Minjian;Hong, Guoqiang;Li, Zhaoxia;Zhu, Zhenyu;Li, Lin
BMB Reports
/
v.38
no.6
/
pp.683-689
/
2005
Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.
Nam, Kyoung Hyup;Choi, Hyuk Jin;Lee, Jae Il;Ko, Jun Kyeung;Han, In Ho;Cho, Won Ho
Journal of Trauma and Injury
/
v.28
no.1
/
pp.9-14
/
2015
Purpose: The aim of this study was to estimate the seropositive prevalence of blood-borne infection in neurotrauma patients who underwent emergent surgical intervention, especially patients with hepatitis B virus (HBV), hepatitis C virus (HCV), syphilis and human immunodefIciency virus (HIV). Methods: A retrospective review identified 559 patients with traumatic brain injury and spinal trauma who underwent emergent surgery between 2007 and 2014. We reviewed the medical records and extracted data, including age, sex, location of lesion, result of serologic tests, time interval of admission and surgery after presenting to emergency room. Serologic tests for HBV, HCV, syphilis and HIV were performed and analyzed to determine whether the seropositive results were confirmed by the surgeon before surgery. Results: The majority of the patients were male (74.6%), and the mean age was $55.4{\pm}20.2years$. Most patients underwent surgery due to traumatic brain injury (90.0%). Fifty-three patients (10.0%) showed a positive result on at least one serologic test. Seropositive rates according to pathogens were 0.5% for syphilis, 5.2% for HBV and 3.9% for HCV. No positive results were noted on the serologic tests for HIV. HBV in patients with spinal cord injury and age from 40 to 49 years were associated with high serologic positive rate, and that result was statistically significant. However, no statistically significant differences were found in the other variables. Serologic results could not confirmed before surgery in the majority of the cases (62.1%), and 10.4% of these patients showed seropositive results. Conclusion: The results of this study emphasize the importance of taking precautions and conducting rapid serologic testing in preventing the occupational transmission of blood-borne viruses to health-care workers.
To assess the contribution of tumor necrosis factor $(TNF){\beta}$ +252 polymorphisms to risk and prognosis of hepatocellular carcinoma (HCC), we enrolled 150 pairs of sex- and age-matched patients with HCC, patients with cirrhosis alone, and unrelated healthy controls. $TNF{\beta}$ +252 genotypes were determined by polymerase chain reaction with restriction fragment length polymorphism. Multivariate analysis indicated that $TNF{\beta}$ G/G genotype [odds ratio (OR), 3.64; 95%CI, 1.49-8.91], hepatitis B surface antigen (OR, 16.38; 95%CI, 8.30-32.33), and antibodies to hepatitis C virus (HCV) (OR, 39.11; 95%CI, 14.83-103.14) were independent risk factors for HCC. There was an additive interaction between $TNF{\beta}$ G/G genotype and chronic hepatitis B virus (HBV)/HCV infection (synergy index=1.15). Multivariate analysis indicated that factors associated with $TNF{\beta}$ G/G genotype included cirrhosis with Child-Pugh C (OR, 4.06; 95%CI, 1.34-12.29), thrombocytopenia (OR, 6.55; 95%CI, 1.46-29.43), and higher serum ${\alpha}$-fetoprotein concentration (OR, 2.53; 95%CI, 1.14-5.62). Patients with $TNF{\beta}$ G/G genotype had poor cumulative survival (p=0.005). Cox proportional hazard model indicated that $TNF{\beta}$ G/G genotype was a biomarker for poor HCC survival (hazard ratio, 1.70; 95%CI, 1.07-2.69). In conclusion, there are independent and additive effects between $TNF{\beta}$ G/G genotype and chronic HBV/HCV infection on risk for HCC. It is a biomarker for poor HCC survival. Carriage of this genotype correlates with disease severity and advanced hepatic fibrosis, which may contribute to a higher risk and poor survival of HCC. Chronic HBV/HCV infected subjects with this genotype should receive more intensive surveillance for early detection of HCC.
Choi, Yujung;Bae, Kil Seoung;Kim, Ki Hwan;Koh, Dae Kyun;Kim, Jong-Hyun
Pediatric Infection and Vaccine
/
v.25
no.2
/
pp.72-81
/
2018
Purpose: This prospective study aimed to investigate the therapeutic efficacy of lamivudine in children with chronic hepatitis B virus (HBV) infection. Methods: During July 2003 through October 2015, children with chronic hepatitis B who visited our institution were included in this study. Fifty-five patients, who received first-line treatment of lamivudine (3 mg/kg, 100 mg maximum) for over three months, were enrolled. After initiating lamivudine, alanine aminotransferase (ALT), HBV-DNA, and HBV markers were followed up at 1 month, 3 months, and every 3 months, thereafter. The treatment endpoint was determined as 1) normalization of ALT, 2) HBeAg seroconversion, and 3) anti-HBe positivity for twelve consecutive months. Results: Thirty-one male (56.4%) and 24 female (43.6%) patients were included. The mean age at treatment initiation was 8.1 years. The mean duration of treatment was 23.4 months. ALT normalization was found in 98.2% (54 of 55). Anti-HBe seroconversion was found in 70.6% (36/51). Loss of HBsAg was found in 10.9% (6/55). All biochemical responses occurred under age seven. The rate of virologic response (defined as HBV-DNA <2,000 IU/mL) at six months after treatment initiation was 78.7% (37/47). At twelve months after reaching treatment endpoint, 87.2% (34/39) maintained their virologic response. Resistance to lamivudine was found in 16.4% (9/55). Conclusions: Lamivudine treatment in Korean pediatric patients with chronic hepatitis B showed better outcomes compared with other studies that implemented similar protocols in foreign populations. Further studies are needed to investigate the efficacy of newly recommended antiviral drugs on the Korean pediatric population.
To estimate the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection and to determine associated risk factors, a population-based seroepidemiologic study was carried out. In 1993, a health examination survey of the population was carried out in rural area known to have a high incidence of liver cancer. The study population were those who volunteered to participate in a health survey over 10 years of age. Examinees were interviewed by specially trained staffs. Sera from 1,033 study subjects were tested for hepatitis B surface antigen (HBsAg) by .everse passive hemagglutinin (RPHA) estimation and for hepatitis C virus antibody (anti-HCV) by 2nd generation passive hemagglutinin (PHA) estimation. The age and sex standardized prevalence of HBsAg was 6.3% which was similar to national average, but that of anti-HCV was 5.1% which was 4 to 5 times higher than that of blood donors or other health examinees in Korea. In a multivariate analysis, transfusion history, surgical operative history, and acupuncture history were not associated with HBsAg positivity. In contrast, acupuncture history (adjusted odds ratio[OR]=2.2 : 95% Confidence interval[CI] 1.0-4.7) and surgical operative history(adjusted OR=2.0 : 95% CI 1.0-4.1) were associated with anti-HCV positivity. The present study suggest that there is an highly endemic area of HCV infection in Korea and probably this endemicity is associated with a parenteral source of HCV infection other than blood transfusion.
Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.
Background: Patients with chronic liver diseases (CLD) may have compromised health related quality of life (HRQoL). Hepatitis B virus (HBV) infection has long been the leading cause of CLD including liver cancer and cirrhosis. Knowledge on different symptom profiles of CLD should help in development of comprehensive treatment and patient care plans. Objective: To access the facets of HRQoL in chronic liver diseases throughout their spectrum of severity. Materials and Methods: A cross-sectional study was conducted in the First Affiliated Hospital of Kunming Medical University in Yunnan Province of China. Both out- and inpatients undergoing treatment protocols for different HBV related liver disease states were consecutively collected from December 2012 to June 2013. ANOVA was used to compare the mean scores of EQ-5D and chronic liver disease questionnaire (CLDQ) among 5 disease groups. The relationship between demographic variables predicting global CLDQ scores and the domains of CLDQ was analysed. Results: A total of 1040 patients including 520 without complications, 91 with compensated cirrhosis, 198 with decompensated cirrhosis, 131 with HCC and 100 with liver failure were recruited. All domains of CLDQ, the means of EQ-5D value and EQ VAS exhibited significant decline with worsening of disease severity from uncomplicated HBV to liver failure. The multivariate regression demonstrated the reduction of mean scores of CLDQ domain at advanced stage. Patients with liver failure and HCC had more HRQoL impairment than other disease states. No effect of patient gender was found. Patient age was associated with 'fatigue' and 'worry' domains (p=0.006; p=0.004) but not with other domains and global scores of CLDQ and ED-5D. Conclusions: The HRQoL in chronic hepatitis B patients is greatly affected by disease states. Care for HBV-related diseases should consider not only the outcomes of treatment strategies but also improvement in patient wellbeing.
Chronic hepatitis B virus (HBV) infection and dietary exposure to aflatoxin B1 (AFB1) are major risk factors for hepatocellular carcinoma (HCC). The aim of this study was to evaluate the role of HBV genetic variation and the R249S mutation of the p53 gene, a marker of AFB1-induced HCC, in Thai patients chronically infected with HBV. Sixty-five patients with and 89 patients without HCC were included. Viral mutations and R249S mutation were characterized by direct sequencing and restriction fragment length polymorphism (RFLP) in serum samples, respectively. The prevalences of T1753C/A/G and A1762T/G1764A mutations in the basal core promotor (BCP) region were significantly higher in the HCC group compared to the non-HCC group. R249S mutation was detected in 6.2% and 3.4% of the HCC and non-HCC groups, respectively, which was not significantly different. By multiple logistic regression analysis, the presence of A1762T/G1764A mutations was independently associated with the risk of HCC in Thai patients.
Dane particle was prepared from the plasma of chronic HBsAg carrier with high levels of HBsAg activity. DNA extracted front Dane particle core after a DNA polymerase reaction with $\alpha$-($^{32}$P) dNTP, was identified as HBV DNA by liquid scintillation counter and agarose gel electrophoresis-G.M. counting. To produce Hepatitis B surface antigen for use as a vaccine against Hepatitis B virus infection, yeast strains harboring recombinant plasmid with Apase promoter was used. Recombinant plasmid was construced from pHBV 130 and pAN 82, transformed into E coli, and then transferred into yeast strains. HBsAg was produced by derepression in Burkholder minimal medium with controlled inorganic phosphate concentration. The kinetics of HBsAg production was also investigated. Total HBsAg activity increased rapidly between 3 and 6 hours after transfer to phosphate-free medium and reached a maximum at around 9th hour. The transfer into phosphate-free medium after 6 hours in high phosphate cell growth medium gave maximum activity.
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