<b>OBJECTIVEb>To analyze the characteristic of T cell response to specific antigen proteins in patients with hepatitis B virus infection.

<b>METHODSb>76 cases were recruited, including four groups, acute hepatitis B (AHB), active phase of chronic hepatitis B (CHB), inactive HBV carriers (AsC) and past HBV infection. T cell responses stimulated by 3 antigen specific proteins of HBV were detected using enzyme linked immunospot (ELISPOT) assay.

<b>RESULTSb>(1) There were no significant difference in frequencies to HBsAg, HBcAg and HBeAg in AHB and CHB. The frequencies to HBsAg and HBcAg in AsC were lower than that to HBeAg, and the frequencies to HBsAg in group of past HBV infection were significantly lower than that to HBcAg and HBeAg. (2) The frequencies to HBsAg in AHB and CHB both were higher than in group of past HBV infection. The frequencies to HBcAg of AHB, CHB and AsC were higher than that of group of past HBV infection. (3) There were no significant difference in magnitude to HBsAg, HBcAg and HBeAg in AHB and AsC. In CHB, the magnitude to HBsAg was lower than that to HBcAg. The magnitude of in group of past HBV infection were HBcAg > HBeAg > HBsAg. (4) In four groups, the sequence of the magnitude to HBsAg from high to low was AHB, CHB, group of past HBV infection and AsC. The magnitude to HBcAg in of AsC was lower than other three groups. As to the magnitude to HBeAg, the difference was no significant between any two groups except between AHB and CHB.

<b>CONCLUSIONSb>The T cell responses in group of AsC to HBeAg were the highest, while the T cell responses to HBcAg were the highest in group of other groups.

Hepatitis B ; immunology ; virology ; Hepatitis B Antibodies ; immunology ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B e Antigens ; immunology ; Hepatitis B virus ; immunology ; isolation & purification ; Humans ; T-Lymphocytes ; immunology

Hepatitis B virus (HBV) infection has been a major global cause of morbidity and mortality. The recognition of the problem led to a worldwide effort to reduce transmission of HBV through routine infant vaccination. HBV infection is the most common cause of chronic liver diseases and hepatocellular carcinoma in Korea. After hepatitis B vaccine era, seroprevalence of hepatits B surface antigen is decreasing, particularly in children. Hepatitis B vaccine is remarkably safe and shows high immunogenicity. Universal childhood immunization with three doses of hepatitis B vaccine in the first year of life is a highly effective method for prevention and control of hepatitis B.
Hepatitis B/*epidemiology/immunology/*prevention & control ; Hepatitis B Antibodies/blood/immunology ; Hepatitis B Vaccines/immunology/therapeutic use ; Hepatitis B virus/genetics/immunology ; Humans ; Vaccination

<b>OBJECTIVEb>Through detecting the standard preparation with series of concentration to indirectly calculate the anti-HBs concentration of the serum samples, a suitable anti-HBs quantitative method for our laboratory was found after comparing the two kinds of methods.

<b>METHODSb>Detecting the anti-HBs standard preparation with series of concentration by RIA method, standard curvilinear equations were obtained by the means of fitting the detected result and the corresponding concentration by log-log model and exponential curve model respectively. Then the fitting efficiency of two curves was compared. By calculating the concentrations of the reference using two standard curvilinear equations, we can compare the accuracy of two quantitative methods.

<b>RESULTb>The error mean square of the exponential curve model is low as 1.2971 and the determinate coefficient is close to 1 with the value of 0.9904. The average concentrations (n=6) of the detected reference calculated by two curvilinear equation with the actual concentration of 30.0 mIU/mL are (32.28 +/- 1.06) and (31.91 +/- 1.06) mIU/ mL respectively. The concentration calculated by exponential curve model is only 6.37% higher than the actual concentration.

<b>CONCLUSIONb>Fitting by exponential curve model is more practical to estimate the actual concentration of the serum samples those will be detected. It can be used as an optimal quantitative method to detect anti-HBs concentration.

Hepatitis B ; blood ; immunology ; virology ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B virus ; immunology ; isolation & purification ; Humans ; Male ; Radioimmunoassay ; methods

<b>OBJECTIVEb>Screening and characterizing high affinity completely humanized single-chain antibody (ScFv) against PreS1 of hepatitis B virus.

<b>METHODSb>A combinatorial library of phage-displayed human ScFv, genes of which were derived from peripheral blood lymphocytes immunized by PreS1 of Hepatitis B Virus in vitro, was constructed. The library contained 7 10(8) clones.

<b>RESULTSb>After 3 rounds panning, a high affinity (K=10(7) to 10(8) mol/L) ScFv specific to PreS1 was obtained. The V(H) belonged to human V(H4) family, and V(1) to V(4) by sequence analysis.

<b>CONCLUSIONb>This study suggests that the method of antigen stimulation in vitro is an expeditious way for the source of human immune antibody. And the ScFv may provide a more satisfactory therapy.

Antibodies, Monoclonal ; biosynthesis ; Antibody Affinity ; Hepatitis B Antibodies ; biosynthesis ; Hepatitis B Surface Antigens ; immunology ; Humans ; Peptide Library ; Protein Precursors ; immunology

<b>OBJECTIVEb>Establish a kind of confirmation method based on ELISA, and use to verify authenticity of HBsAb + in HBsAg + HBsAb + serum, pick and get rid of the false masculine gender the result, and avoid the mistake diagnosis.

<b>METHODb>Collect 60 pieces of serum whose thick degree of HBsAg at 1000 COI above tested by ECLIA as confirm serum, mixed the confirm serum of different dilution with HBsAb positive serum to screen and verify best thick degree of HBsAg. Collected 40 pieces of HBsAg + HBsAb + serum, ELISA tested the descend rate of HBsAb COI after neutralized with confirm serum in order to confirm authenticity of HBsAb + in pieces of HBsAg + HBsAb + serum.

<b>RESULTb>When thick degree of HBsAg is 2000 COI, the performance of neutralization to HBsAb is best. The ELISA confirmatory test is fully consistent with the ECLIA method with true positive of 37 pieces of HBsAg + HBsAb + serum while false-positive of 3 pieces of serum.

<b>CONCLUSIONb>The ELISA confirm method is a simple, accurate and low cost initial validation method.

Enzyme-Linked Immunosorbent Assay ; methods ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; immunology ; Humans

<b>OBJECTIVEb>To investigate the positive ratio and clinical significance of PreS1Ag and anti-PreS1 in patients with chronic hepatitis B.

<b>METHODSb>428 patients with chronic HBV infection were collected, these patients were divided into e antigen-positive CHB group, e antigen-negative CHB group, inactive HBsAg carrier group and HBsAg serum conversion group. The difference of positive ratio of PreS1Ag and anti-PreS1 among all groups or between every two groups were analyzed; The relationship of PreS1Ag and anti-PreS1 with HBV M and HBV DNA were also analyzed. SPSS13.0 software was used for statistical treatment. Fourfold table chi-square test or matched-pairs chi-square test was used for enumeration data, and independent sampler t test or rank-sum test was used for measurement data.

<b>RESULTSb>The differences of PreS1Ag among four groups were statistically significant (X2=141.7, P<0.05). The positive ratio of PreS1Ag in e antigen-positive CHB group was 95.7%, followed by 82.8% in e antigen-negative CHB group, 13.2% in inactive HBsAg carrier group and 2.2% in HBsAg serum conversion group. The difference of positive ratio of anti-PreS1 between HBsAg seroconversion group and HBsAg positive group was statistically significant (X2=6.919, P<0.05), which indicated that anti-PreS1 had good correlation with HBsAg seroconversion. The average absorbance ratio of PreS1Ag in high viral replication group (179.30) was higher than that in low viral replication group (133.87), statistical significance appeared (Z=-3.86, P<0.05). Though the difference of absorbance ratio of anti-PreS1 between two groups had no statistical significance (P>0.05), descent trend was apparent with virus replication level ascending. We analyzed the concordance of anti-HBs and anti-PreS1 by matched-pairs chi-square test, result showed no statistical significance of detection rate between them, X2=0.262, P>0.05. Serum PreS1Ag, HBeAg or HBcAg in liver tissue in reflecting hepatitis B replication had correlation with HBV DNA (X2=33.840, 24.159, 4.854 in order, P<0.05). Correlation coefficient between PreS1Ag and HBV DNA was higher (r=0.628) than that between HBeAg and HBV DNA (r=0.563).

<b>CONCLUSIONb>PreS1Ag was more sensitive than HBeAg in diagnosing viral replication in patients with chronic hepatitis B. Anti-PreS1 as protective antibody may be involved in clearance of hepatitis B, positive result indicated recovery of chronic hepatitis B.

Adult ; Female ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Male ; Middle Aged ; Protein Precursors ; blood ; immunology

<b>OBJECTIVEb>The aim of the study was to evaluate the anti-HBs persistence and the long term preventive efficacy after vaccination 23 years with plasma-derived hepatitis B vaccine.

<b>METHODSb>The study consisted of 261 children who were 5 - 9 years aged, from two primary schools in two townships of Xi'an. 126 children were randomly selected as vaccine group, and 135 children in control group. These children were followed up again in 2009. Excluding self-inoculation, the vaccine and control groups were 81 and 75, who was used to ask to recall details of their experience for vaccination and liver-related illnesses during past twelve years. Individuals who had anti-HBs titers less 10 mIU/ml, HBsAg, anti-HBc and HBV-DNA all were negative, were given a booster dose vaccine and retest for anti-HBs titer after one month.

<b>RESULTSb>After eliminated the interference of an early booster dose and vaccination outside the study, the positive rate of anti-HBs was 48.1% (39/81) in the vaccine group at year 23, higher than 34.7% (26/75) in control group. At year 23 after primary vaccination, 84.0% (21/25) individuals in the vaccine group whose anti-HBs and anti-HBc both are negative showed a stronger anamnestic response after received a booster dose, while 7.5% (3/40) in the control group. At year 23 after primary vaccination, none clinical case of hepatitis B was found among 194 individuals. However, anti-HBc positive rate in the vaccine group was 16.0% (13/81), while the rate in the control group was 30.7% (23/75) (χ(2) = 4.687, P < 0.05).

<b>CONCLUSIONb>At 23 years after implemented a full course of plasma-derived hepatitis B vaccine, the recipients of vaccine were maintained anti-HBs at a high level or strong immunological memory.

Child ; Child, Preschool ; Follow-Up Studies ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; immunology ; Humans ; Immunization, Secondary ; Immunologic Memory ; immunology ; Plasma ; immunology

<b>OBJECTIVEb>To analyze the information of the supplementary card for hepatitis B and the laboratory confirmed result of immunoglobulin M antibody to hepatitis B virus (HBV) Core Antigen (anti-HBc IgM) for the suspected acute hepatitis B to evaluate the hepatitis B report data on pilot surveillance.

<b>METHODSb>200 counties were established in China for hepatitis B pilot surveillance and 63 641 cases were reported. We added a supplementary card in National Notificable Disease Reporting System (NNDRS) and all the reported hepatitis B cases in NNDRS were required to fill the supplementary card. Venous blood 5 ml was collected and a confirmed test of anti-HBc IgM was made for suspected acute hepatitis B. We made confirmed diagnosis for the suspected acute hepatitis B according to the supplementary card information of the reporting card and the confirmed test result of anti-HBc IgM.

<b>RESULTSb>63 641 hepatitis B cases were reported in 200 hepatitis B pilot surveillance counties in 2013. Among 1 723 cases which were filled with the HBsAg positive within six months in supplementary card, 735 cases were reported as chronic hepatitis B, the proportion was 42.66%. Among 4 582 cases which were filled with anti-HBc IgM positive in supplementary card, 2 436 cases were reported as acute hepatitis B, the proportion was 53.16%. 1 829 cases were reported as chronic hepatitis B, the proportion was 39.92%. The validity cases of the information for liver puncture and the HBV surface antigen (HBsAg) transform during the recovery period in supplementary cards for all the reporting cases were 579 and 4 961, and the rate were 0.91% and 7.80%, respectively. 4 302 suspected acute cases were made confirmed diagnosis, and 1 197 cases (27.82%) were confirmed acute and 2 590 cases (60.20%) were confirmed chronic.

<b>CONCLUSIONb>Clinical doctors failed to make full use of the information of supplementary cards to make classification diagnose for hepatitis B. Suspected acute hepatitis B with anti-HBc IgM positive should be pay attention to follow up and further distinguish acute or chronic hepatitis B according to the HBsAg transform.

China ; epidemiology ; Hepatitis B ; epidemiology ; Hepatitis B Antibodies ; blood ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Humans ; Immunoglobulin M ; blood ; Sentinel Surveillance

<b>OBJECTIVEb>To see the HBV DNA detection instance in the HBsAg negative people and to study the serological method detection strategy for detecting hepatitis B virus large surface protein (HBLP) to filtrate the occult HBV infection.

<b>METHODSb>The HBsAg negative serum samples were divided into HBsAb negative and positive two species according to the hepatitis B virus markers (HBVM) in daily work excepting the special HBVM modes. Total 2000 stochastic serum samples with 1000 HBsAb negative results and 1000 HBsAb positive results were collected from the copy tubes to detect HBVM with national ELISA reagent kits and put them -20 degrees C frostily. Mixed samples (8 x 30 microl) were analyzed with fluorescence quantitative PCR (FQ-PCR) and filtrated the individual positive samples. The filtrated samples were doubly tested again with American MONOLISA HBsAg ULTRA reagents.

<b>RESULTSb>No HBV DNA positive results were found out from the 1000 HBsAb positive samples and 19 cases HBV DNA positive results were found out from the 1000 HBsAb negative samples. On these 19 samples, the HBsAg results from the American MONOLISA HBsAg ULTRA reagents were all positive and the HBLP results were all positive, too. The 19 HBV DNA quantitative results were divided into 2 cases more than 500 copies/ml, 3 cases between 400-500 copies/ ml, 3 cases between 300-400 copies/ml, 7 cases between 200-300 copies/ml and 4 cases between 100-200 copies/ml.

<b>CONCLUSIONb>The leaked samples tested HBsAg with national reagents are mostly from the HBsAb negative people. HBLP results may be positive on these samples and detecting HBLP marker is propitious to filtrate the occult HBV infection. This study provided a kind of serological reference for actively searching for the detecting strategy in occult HBV infection field.

Antibody Specificity ; Hepatitis B ; diagnosis ; Hepatitis B Antibodies ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B virus ; isolation & purification ; Humans ; Laboratories ; Polymerase Chain Reaction

Hepatitis B ; immunology ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; immunology ; physiology ; Humans ; Immunologic Tests ; methods ; Virus Replication