• Journal of Preventive Medicine and Public Health
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• v.19 no.2 s.20
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• pp.281-292
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• 1986

Infection by hepatitis B virus is one of the major health problems of this nation. HBsAg positive rates of general population and school children were known to be as about 8 percent and 3.9 to 5.9 percent respectively. To study the incidence rate of hepatitis B infection in school children of rural area, author had examined 475 school children of relatively isolated agricultural area for baseline prevalence of hepatitis B virus serologic markers and followed up 415 school children during 10 months to determined the frequency of serologic conversion. The major results are summarized as followings: 1) Among the 278 susceptible children who were followed up, 26 had seroconversion for HBsAg or Anti-HBs. Therefore, the cumulative incidence rate during 10 months is estimated 9.4%. 2) The incidence rate of hepatitis B infection tends to increase with age (6-9yrs: 3.2%, 10-14yrs: 9.5%, 15-17yrs: 18.9%), and the incidence rate in male (13.0%) was higher than in female (5.7%). 3) The incidence rates of hepatitis B virus infection were not different statistically among three economic classes (The rates of lower, middle and higher class were 11.8%, 7.1% and 10.5%.). 4) The incidence rates of hepatitis B virus infection were not different statistically between visitors and non-visitors of clinic or hospital, dental clinic, persons received IV and not received IV, and persons with familial history and without familial history of liver diseases. Therefore all of these factors were not identified as risk factor of hepatitis B virus infection. And the transmissibility within the class of school was not recognized, too. 5) Among the 25 children who were HBsAg positive when enrolled, 15 (60%) were still HBsAg positive, who were identified as chronic carrier 15 of 415 school children were chronic carriers, then chronic carrier rate was estimated 3.6%, and there was no difference between sexes. 6) Of 38 children who had been Anti-HBs positive when enrolled, 5 (13.2%) lost Anti-HBs. Therefore, the loss rate of Anti-HBs per year is estimated to be 15.8%.

• Journal of Life Science
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• v.15 no.1 s.68
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• pp.38-44
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• 2005

Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV, there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-1c1c7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-a, the inducibility of TNF-a expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-a expression. Infection of Hepa-1c1c7 cells with the serum of HBV carrier also increased TNF-a mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in $\beta-actin$ mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-lc1c7 in vitro and the viral infection induces TNF-a expression, which suggests that Hepa-lc1c7, a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.

• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.639-648
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• 1997

Background : Arterial hypoxemia has been noted in patients with liver cirrhosis because of bronchial vessel dilatation. Cabenes et al. reported that bronchial hyperresponsiveness to the metacholine inhalation was observed in patients of left side heart failure, he suggested that one of the mechanism was bronchial vessel dilatation. We hypothesized that patients of liver cirrhosis might have bronchial hyperresponsiveness to metacholine inhalation due to portal hypertension. We evaluate the relationship between bronchial responsiveness and severity of liver cirrhosis, severity of portal hypertension. Methods : In the 22 patients of the liver cirrhosis with clinical portal hypertension, metacholine provocation test was done and determined $PC_{20}FEV1$. We classified liver cirrhosis according to Pugh-Child classification. Esophagogastroscopies were performed for the evaluation of the relationship between bronchial hyperresponsiveness and severity of esophageal varix. Results : In the 22 cases of the liver cirrhosis with clinical portal hypertension. The causes of liver cirrhosis, alcoholic hepatitis was 9 cases, hepatitis B virus was 12 cases, hepatitis C virus was 1 case, and 151 cases (68.18%) of total 22 cases were positive in metacholine provocation test. In positive cases. There was no significant relationship between $PC_{20}FEV1$ and severity of liver cirrhosis which were classified by Pugh-Child classification or severity of esophageal varix(p<0.05). Conclusion : we observed that bronchial responsiveness to metacholine increased in the patients of liver cirrhosis and there was no significant relationship between the severity of liver cirrhosis and the severity of esophageal varix.

• Journal of Preventive Medicine and Public Health
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• v.33 no.3
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• pp.323-333
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• 2000

Objectives : Chronic HBsAg carriers are the principal source of infection for other susceptible people, and are themselves at high risk of developing serious liver diseases. In Korea, it has been estimated that 65-75% of the HBsAg positives remained as persistent carriers. Additionally, familial clustering of MBV infection has frequently been observed among carriers. Some would become progressive, chronic hepatitis patients, and others would not. The aim of this study was to evaluate the association between various factors, such as the duration of infection, type of virus, mutation of precore/core region in HBV, major histocompatibility class-I, and developing chronic liver diseases among familial HBV carriers. Methods : Chronic carrier status was identified by repeated serological tests for HBsAg at intervals of six months or more. A familial chronic carrier was defined when the disease was observed in a family member over two generations. Two families were recruited, among which a total of 20 chronic HBsAg carriers(11 carriers in No.1, and 9 in No.2 family) were identified. Data on the general characteristics and liver disease status were collected. Identification of the HBV-DNA was successful only for 13 subjects among the 20 carriers. Analysis of viral DNA in terms of subtype, pre-core and core region mutations was carried out. The type of major histocompatibility class-1 for the 13 subjects was also analysed. Results & Conclusions : Seven of 10 chronic HBV carriers of the 1st generation and one of 10 of the 2nd generation were clinical patients with chronic hepatitis, the others, three of the 1 st and nine of the 2nd generation, were asymptomatic carriers. This data indicates that the duration of HBV carriage is one of the major factors for disease severity. The subtype of HBsAg analysed using MBV-DNA identified in 13 carriers were adr, and the pattern of precore nonsense mutation in HBV-DNA was identical among family members, which meads that the same virus strains were transmitted between the family members. The association between the precore or core mutations in HBV-DNA and the disease severity was not observed. While it was suggested that a specific type of MHC class-I may be related to disease progression.

• IMMUNE NETWORK
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• v.4 no.1
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• pp.16-22
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• 2004

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.

• Journal of Preventive Medicine and Public Health
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• v.21 no.1 s.23
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• pp.89-98
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• 1988

While there have been not a few reports on the seroepidemiological characteristics of hepatitis B virus (HBV) infection in Korea, most of them, however, have had several limitations; operational definition of HBV infection, validity of detection methods of HBV serologic markers, size of the study population, and confirmation of the vaccination history against HBV, etc. In order to avoid such limitations, authors randomly selected 1,495 healthy adults among the 217,511 insured (target population) of Korean Medical Insurance Corporation, living in seoul, and tested HBV serologic markers by RIA method and conducted direct interview to them. Although HBV serologic markers (HBsAg, anti-HBs and anti-HBc) of all the subjects were tested, 392(26.2%) of interview failure cases and 361 vaccinee were excluded from the actual population. Finally, the serologic markers tested of 742 nonvaccinee (study population) only were analysed for the seroepidemiologic observation of the natural infection of HBV. The seroepidemiological characteristics of HBV infection in Korea were as follows ; 1. Point prevalence of HBs antigenemia was 11.7(9.1{\sim}14.3)% in male, which was slightly higher than that of female, 9.5($3.7{\sim}15.3$)%. This level was one of the highest among those of Asian-Pacific countries. Decreasing tendency of HBsAg prevalence alter the age of 50 was observed, which seems to be due to selective attrition of HBV chronic carriers among the healthy adults and/or to the limited-lasting duration of the HBs antigenemia, in part. 2. Point prevalence of anti-HBc(78.8% in male,50.9% in female) was higher than that of anti-HBs(65.2% in male,46.6% in female), respectively. And both of them were higher in male than in female. Increasing tendency of the prevalence of both antibodies was observed by age, which seems to be largely due to recurrent infection in adults and to some cumulative effect, in part, of their relatively longer-lasting duration. 3. The level of HBV infection defined by positive for at least one of the 3 serologic markers of HBV by RIA method was 84.7($81.8{\sim}87.6$)% in male and 61.2($51.9{\sim}70.5$)% in female, which was also one of the highest among those of Asian-Pacific countries. The proportion of susceptible population to HBV infection among healthy adults was 15.3% in male and 38.8% in female. 4. The relative frequency of current or past infection and chronic carrier among HBV infected person was estimated. The currently or past infected was estimated 75.7% in male and 71.8% in female, and chronic carrier state, 13.8% in male and 14.1% in female. The analysis of the geometric mean of the antibody titer in anti-HBs positive sera indicated also to be compatible with the above findings, suggesting that active, even though inapparent, infection of HBV occur so frequently among healthy adults in Korea.

• Journal of the Korean Society of School Health
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• v.15 no.2
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• pp.183-191
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• 2002

This study is aimed at offering basic data about prevention of Hepatitis B virus and infection control to dental hygiene students who will work mostly at dental offices which are characteristically exposed to a high risk of HBV infection. For this study, surveyed were 188 sophomores and juniors of the dental hygiene department who volunteered to undergo the tests of HBsAg and HBsAb. The examination of the blood collected from the samples and analysis of their perception about hygiene resulted in following conclusion: Positive HBsAg Four was found in 4 testees(2.1%) while 118(62.8%) revealed positive HBsAb. Juniors showed higher positive rate in HBsAg whereas sophomores had higher positive rate in HBsAb. But such difference has statistically no significance. Generally, students seemed to pay little attention to HBV, judging from the survey of their perception about the present state of HBsAg and HBsAb(p>0.05), conduct of infection(p<0.05), completion of 3 requested vaccinations(p>0.05), and formation of antibody(p>0.05). With regard to the infection routes of HBV, most students(92.4%) replied "through blood", which is statistically insignificant, though. Next ratio goes to the reply "through contaminated injectors". This reply came more from sophomores than from juniors, a difference which is statistically significant(p<0.05). The lowest rate of possible infection(29.2%) was thought to be "through breast-feeding of a positive mother"(p<0.05). In general, it turned out that sophomores had more knowledge about the infection routes of HBV than juniors. In terms of clinical history among family members, 6(3.1%) reported that some of their family members are currently suffering from a liver complaint, 3(1.6%) replied their family members were once afflicted, and 4(2.1%) said their members died of hepatitis. Except 10(4.7%), all the surveyees replied that their states of health are better than normal. Generally, sophomores are healthier than juniors except for the very health case, a difference which is statistically of no value.

• IMMUNE NETWORK
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• v.3 no.2
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• pp.126-135
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• 2003

Background: One of the important factors in the prognosis of chronic hepatitis B patient is the degree of replication of hepatitis B virus (HBV). It has been known that HBV DNA polymerase plays the essential role in the replication of HBV. HBV DNA polymerase is composed of four domains, TP (Terminal protein), spacer, RT (Reverse transcriptase) and RNaseH. Among these domains, tyrosine, the $65^{th}$ residue of TP is an important residue in protein-priming reaction that initiates reverse transcription. If monoclonal antibody that recognizes around tyrosine residue were selected, it could be applied to further study of HBV replication. Methods: To produce TP-specific scFv (single-chain Fv) by phage display, mice were immunized using synthetic TP-peptide contains $57{\sim}80^{th}$ amino acid residues of TP domain. After isolation of mRNA of heavy-variable region ($V_H$) and light-chain variable region ($V_L$) from the spleen of the immunized mouse, DNA of $V_H$ and $V_L$ were obtained by RT-PCR and joined by a DNA linker encoding peptide (Gly4Ser)3 as a scFv DNA fragments. ScFv DNA fragments were cloned into a phagemid vector. scFv was expressed in E.coli TG1 as a fusion protein with E tag and phage gIII. To select the scFv that has specific affinity to TP-peptide from the phage-antibody library, we used two cycles of panning and colony lift assay. Results: The TP-peptide-specific scFv was isolated by selection process using TP-peptide as an antigen. Selected scFv had 30 kDa of protein size and its nucleotide sequences were analyzed. Indirect- and competitive-ELISA revealed that the selected scFv specifically recognized both TP-peptide and the HBV DNA polymerase. Conclusion: The scFv that recognizes the TP domain of the HBV DNA polymerase was isolated by phage display.

• Clinical and Experimental Pediatrics
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• v.50 no.4
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• pp.348-354
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• 2007

Purpose : Perinatal hepatitis B viral infection is decreasing; however, 10% of babies to HBeAg positive mothers still become chronic carriers despite perinatal prophylaxis. Although, the cause of prophylaxis failure is still unclear, an importance of maternal HBV-DNA level at the delivery time has been suggested. This study was established to certify if it would be a useful predictable factor for the outcome of perinatal prophylaxis. Methods : Twenty-nine HBeAg positive mothers whose babies had known outcomes of prophylaxis were selected. To determine the amount of maternal HBV-DNA, a quantitative PCR was performed with the WHO International Standard for HBV DNA NAT assays. Results : The mean logarithm HBV-DNA level of mothers with failed outcomes was significantly higher than that of mothers with succeessful outcomes (7.99 vs. 6.72, P=0.015). The predictable maternal HBV-DNA cut-off level to prophylaxis outcome was $2.83{\times}10^7copies/mL$ (100 pg/mL). None out of the case 16 (0%) who had below this level, and 5 out of 13 (38.5%) who had above this level of maternal HBV-DNA failed in perinatal prophylaxis. Conclusion : Mothers with higher levels of HBV-DNA at delivery time would be prone to a worse outcome of prophylaxis using the conventional approach. Perinatal prophylaxis failure rate can be reduced, if we try to introduce more potent prophylactic treatment into the cases with this risk factor.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.5 no.1
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• pp.39-50
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• 2002

Hepatitis B viral infection which affect about 10% of Korean population manifests asymptomatic carrier, chronic hepatitis and liver cirrhosis and even associates with hepatocellular carcinoma. Clinical manifestations induced by hepatitis B virus vary depending on the degree of immune response by cytotoxic T cells against viral epitope-presenting liver cells. Since hepatitis B virus presents high rate of mutaton that might change the presented epitope and eventually alter immune response, viral mutations, especially in promoters and enhancers, have an important implication in hepatic inflammation and viral replication. To identify mutations related to the hepatic inflammation, we investigated sequence variations of hepatitis B viral promotor regions in the presence or absence of symptoms in hepatitis B carriers. For this, sera from persistently hepatitis B virus-infected mother-child pairs were collected. After PCR amplifiation of all hepatitis B viral promoters (C promoter, S1 promoter, S2/S promoter, X promoter) using serum DNA from each pair, viral promotors were sequenced by automatic sequencer and then sequence data were analyzed by ClustalW. In most cases, the dominant type of maternal virus was transmitted to the child. However, in some children, some new host specific viral variants could be observed in Cp, S1p and S2/Sp. The mutations in C promoter did not seem to be vertically transmitted but arose in new host independently after the wild type had been transmitted. Enhancer I containing X promoter revealed high host specific variations as has been reported before. Two S promoters, S1p and S2/Sp, have shown some point mutations in children, but no deletion mutations were detected as in chronic hepatitis patients in whom deletion mutations are frequently found. In conclusion, the children with the vertically transmitted hepatitis B virus mostly retain the dominant type virus that had been transmitted. However, host specific variants tended to accumulate over time, possibly as clinical symptoms develop.