Alterations in the human immune response to the hepatitis B vaccine among the elderly

The specific binding of hepatitis B (HBs) antigen by lymphocytes from old people immunized with hepatitis B vaccine was explored. For that purpose HBs antigen was combined with fluorescent microspheres, and labeled antigen was allowed to react with lymphocytes from HBs vaccine-responsive or unresponsive people. Lymphocytes from 10 responders and 14 nonresponders were tested for their antigen-binding ability. For controls, lymphocytes were incubated with microspheres bearing human albumin. Lymphocytes from 8 out of 10 responders were able to recognize HBs antigen; for the nonresponders the ratio was 9 out of 14. HBs-binding lymphocytes were B cells but not T lymphocytes. B and T cells from responders and nonresponders were combined and cultivated for 8 days in the presence of HBs antigen, and antibody-producing cells were counted. Neither B cells alone nor B cells plus T cells from nonresponders were able to produce antibody. On the other hand B cells from unresponsive old people produced antibodies when they were cultivated in the presence of HBs antigen and T cells from responsive old people. These data suggest that some elderly individuals who do not produce antibody after in vivo immunization by HBs vaccine do have antibody-producing cells. Instead of a gap in their immune repertoire, these people are suffering from immune dysfunction.     

IgE antibody against hepatitis B surface antigen in mice

Hepatitis B surface antigen (HBs antigen) was examined for elicitation of IgE production by injection into mice. The Prausnitz-Küstner (PK)-type skin test in the rat was employed for detection of IgE antibody to HBs antigen, because no sufficient purified HBs antigen was available as the challenging antigen for the passive cutaneous anaphylaxis (PCA) test in rats. The positive PK test was considered to be due to IgE antibody, since the active principle was inactivated by heating the sera at 56 C for 30 min, did not bind to protein A and was eliminated by anti-mouse IgE antisera. These data indicate that the PK-type test in rats can be used for detection of mouse IgE antibody when the amount of a test sample is not sufficient for the PCA test in rats.     

Detection of HBs antigen with latex particles coated with human antibody prepared by affinity chromatography on concanavalin A-sepharose.

Human HBs antibody was isolated by affinity chromatography on HBs antigen absorbed to concanavalin A linked to Sepharose 4B. When a human anti-HBs immunoglobulin preparation obtained by Cohn's cold ethanol fractionation method was used as a starting material, the antibody was concentrated about 10 times in terms of the passive hemagglutination titer with a recovery rate higher than 50%. Latex particles coated with human anti-HBs antibody thus prepared were proved to be useful in detecting HBs antigen in human blood samples. In its sensitivity and in rapidity of its performance, the antibody-coated latex agglutination test seems to be superior to conventional immunodiffusion techniques.     

HBs antigen and antibody in Barcelona]

The frequency of HBs antigen and antibody has been studied among three populations living in Spain: blood donors, hospital staff, renal deficient hemodialyzed patients. Total results and those given with reference to age, sex, place of birth and residence are compared to each other and to those obtained for populations living in Languedoc.     

乙肝病毒表面抗原和抗体双阳性者中病毒S区基因序列分析

为了解乙肝病毒 (HBV)表面抗原和抗体双阳性患者中病毒的基因型及其HVBS区是否有变异。用放射免疫试剂检测HBsAg阳性样品中的抗 HBs抗体 ,用聚合酶链反应法检测双阳性样品中的HBVDNA ,然后对阳性样品进行克隆和基因序列分析 ,并将所得序列与HBV不同基因型的代表株进行比较分析。结果显示 389例HBsAg阳性样品中有 10例为抗HBs抗体阳性 ;该 10例双阳性样品中有 5例为HBVDNA阳性 ;序列分析显示该 5株HBV均为B基因型 ,其中 4株为adw亚型 ,1株为adr亚型 ;其中有 2株在S区的“a”决定簇的氨基酸发生了变异     

Comparison of hepatitis B,core, HBc,and hepatitis B antibody,anti HBs,in a presumed low risk donor population

Donors screened by medical social history interview negative for high risk behavior or communicable disease history, but subsequently exhibiting reactive serological markers, emphasize importance of duel safe guarding factors for determining donor suitability. This report examines a relationship between two immunoabsorption assay tests, hepatitis B core (HBc) antibody, a required food and drug administration (FDA) test, and hepatitis B antibody (anti HBs), non-required test. Reactive serology results, 129 cases, 3,581 donors (2008–2012) for HBc as the only initially positive serological marker were subjected to anti HBs testing in this history pre-screened donor population. Enzyme linked immunoabsorption assay kits hepatitis B, core and antibody, were used in this study. All samples were initially tested for human immunodeficiency virus, hepatitis B, and hepatitis C, utilizing nucleic acid testing and antigen antibody immunoabsorption assay. Testing was performed by a FDA-registered CLEA-certified reference laboratory. Samples were deceased donor blood samples and a limited number of pre-mortem samples, separated, stored and analyzed according to manufacturer recommendation and FDA regulations. 129 reactive HBc only samples, were subsequently tested for anti HBs. Of these 129, 94 were found to be reactive for anti HBs. This represented 72 % of samples tested for antibody, a higher percentage than anticipated for a medical history negative, low risk population.     

Comparative study on subtypes of hepatitis B surface antigen (HBs Ag) in two regions of southern Japan, Okinawa and Kumamoto. Prevalence and unusual subtypes

A total of 209 hepatitis B surface antigen (HBs Ag)-positive sera from two possibly ethnologically different regions of southern Japan, Okinawa and Kumamoto, were subtyped by counterelectrophoresis using monospecific antisera to a, d, y, w, and r antigenic determinants. There was a marked difference in the prevalence of the w and r determinants in the two regions; the frequency of the HBs Ag/adr subtype was 95% in Kumamoto and 47% in Okinawa, suggesting that HBs Ag subtypes may be one of the markers reflecting an ethnological difference between the peoples of the two regions. Seven and three serum specimens in Okinawa and Kumamoto respectively showed unusual HBs Ag subtypes: four carried adwr (two specimens), aywr, or adyw determinants, five had only a, and the remaining one carried ar.     

The role of immigration in the geographic distribution of the HBs antigen]

The study of HBs antigen frequency among almost 130 000 donors,--according to their place of birth--,shows a significantly higher frequency among foreign residents. The general frequency represents the arithmetic average between that of natives and that of foreign residents. The distribution of ad and ay subtypes is analogous to that observed in the different native countries.     

e antigen in acute hepatitis B.

To examine the association between e antigen and hepatitis-B surface antigen (HBs Ag) we studied 90 inpatients with acute viral hepatitis type B. e Antigen was present in 24 of the patients; these patients had detectable levels of HBs Ag for significantly longer than the 66 with no e antigen in their serum. The HBs Ag subtypes D (adw) and Y (ayw) were similarly distributed among patients with e antigen and among those without, and no differences in the results of biochemical liver function tests were observed between the two groups during the acute phase of illness. Three of the five patients who developed clinical and histological signs of chronic liver disease were positive for e antigen, a finding which supports the hypothesis that e antigen has a prognostic value in hepatitis B.     

A simple purification procedure for hepatitis B surface antigen (HBs) by monoclonal antibody immunosorbent affinity chromatography

Using immobilized monoclonal antibodies (anti-HBs) to hepatitis B surface antigen (HBs) as an immunosorbent affinity column, a simple and effective procedure for HBs purification has been developed. A serum sample containing high titer HBs (i.e. HBe-positive serum) is passed through the column without prior treatment. The HBs is further purified by cesium chloride gradient ultracentrifugation. The recovery of HBs is greater than 70% while the purity is very comparable to those obtained from several isopycnic and rate zonal ultracentrifugation procedures (Dreesman et al., 1972). Over a period of two months, the column was used repeatedly for thirty cycles without any noticeable deterioration.     

Age and sex of chronic carriers of HBs antigen among 130,000 blood donors]

The frequency of HBs antigen, using radioimmunology is studied in blood donors from Languedoc (70.894 men and 60.760 women are tested, between 18 and 60 years old). The frequency in man is found to be 0,6 %, except between 20 and 30 years whereas it reaches 1,0 %; the frequency in woman is 0,4 % except between 18 and 36 years where it is 0,4 %.     

Effect of ploidy and the status of the MAT locus in Saccharomyces cerevisiae yeast cells on the mitotic stability of episomal plasmids and the level of expression of the HBsAg gene of hepatitis B virus]

The frequency of the YEp13, pEF91, YEp13 + HBs, pNMVG3954 plasmids' elimination in a series of isogenic strains n. 2n and 3n was studied. Plasmid stability and the level of expression of the gene of hepatitis B surface antigen (HBsAg) increased in polyploid transformants as a result of increase in plasmid copy number. Heterozygotes MATa/MAT differed from the homozygotes in higher stability of YEp13, pEF91, and YEp13 + HBs plasmids having the same quantity of the HBsAg antigen. The appearance of negative properties--destabilization of episomal plasmids or decrease in synthesis of heterologous protein have been discovered in certain diploid cell. These results point to limitations in constructing a polyploid producer on the basis of the similar type of genome.     

Hepatocellular carcinoma in Japan and its linkage to infection with hepatitis C virus

Hepatitis B and C viruses are closely related to the development of liver cirrhosis and hepatocellular carcinoma. Recently these two viral agents were found to be the major causative agents of hepatocellular carcinoma in Japan. An increase in the number of HCV antibody-positive patients, but a decrease in the number of HBs antigen-positive patients with hepato cellular carcinoma has been noted over the last 15 years. In the late 1980s about 70% of the patients with hepatocellular carcinoma were found to be positive for HCV antibody and 24% for HBs antigen. Presence of several subtypes of HCV was reported. Hypervariable region (HVR) in a putative envelope, gp70, was detected within the same subtype. Variability of HVR seemed to be the result of spontaneous mutation caused after infection. Such changes with time in the sequence of the HCV genome in the blood of patients with type C hepatitis are likely to be due to immunological surveillance by the host.     

Pre-s1 antigen-dependent infection of Tupaia hepatocyte cultures with human hepatitis B virus

The susceptibility of the tree shrew Tupaia belangeri to human hepatitis B virus (HBV) has been demonstrated both in vivo and in vitro. In this study, we show that purified HBV infects primary T. belangeri hepatocyte cultures in a very specific manner, as detected by HBV covalently closed circular DNA, mRNA, HBV e antigen, and HBsAg production. A monoclonal antibody (MAb), MA18/7, directed against the pre-S1 domain of the large HBs protein, which has been shown to neutralize infectivity of HBV for primary human hepatocytes, also blocked infection of primary Tupaia hepatocytes. MAbs against the pre-S2 domain of HBs inhibited infection only partially, whereas an S MAb and polyvalent anti-HBs antibodies neutralized infection completely. Thus, both pre-S1 and S antigens are necessary for infection in the tupaia. Using subviral particles, >70% of primary Tupaia hepatocytes are capable of specific binding of pre-S1-rich HBsAg, showing localization in distinct membrane areas. The data show that the early steps of HBV infection in Tupaia hepatocyte cultures are comparable to those in the human system.     

Hepatitis B core and surface antigens and delta agent in chronic liver disease in Kuwait

Hepatitis B surface antigen was detected immunohistochemically in 25 out of 85 liver biopsies (29.4%) of chronic liver disease. Core antigen was also demonstrated in 9 of the 25 Hepatitis HBs Ag positive biopsies (36%). Delta agent however, was found in only one case of HBs positive chronic active hepatitis. The number of hepatocytes staining positively for HBc antigen was greater in those biopsies with the strongest staining for HBs antigen. The only case of chronic active hepatitis positive for delta agent showed that the positive staining was confined to the nuclei of few hepatocytes. The routine histology showed chronic active hepatitis with a moderate degree of inflammation. The present results confirm our previous reports that almost one third of chronic liver disease in Kuwait is associated with hepatitis B infection. Previous serological studies suggest that delta agent infection is also common; however, the present study suggest that delta agent may be a transient, and not a major, contributing factor in the progression of liver disease.     

Prevalence of HBsAg and anti HBs in hospital personnel

Of the 473 employees of a Prague hospital, examined for the presence of hepatitis B surface antigen (HBsAg) and the antibody to it (anti HBs), 23 (4.9%) were shown to have the antigen and 88 (18.6%) to have the antibody. No differences were found in the prevalence of HBsAg and anti HB according to sex and age, but the percentage of persons with antibodies increased with increasing length of occupation. Significantly higher levels of antibodies were found in persons having direct contact with the patients or their excretions (nurses, laboratory assistants). Positive serological findings were more frequent in employees with a history of falling ill with hepatitis after entering employment in the health services than in persons without this history. In the serological evidence of hepatitis B according to work place, those working in biochemical and haematological laboratories and in the blood transfusion department took first place, followed by persons employed in medical and surgical departments.     

多聚嘧啶序列结合蛋白（PTB）与HBV转录后调节元件（HPRE）结合抑制HBV表面抗原的基因表达

采用RT-PCR验证PTB与HPRE在体外的特异性结合。用HepG2.2.15细胞系、HBs-HPRE瞬时转染Hela细胞探讨PTB对HBV基因表达的影响。结果显示PTB能与HPRE特异性地结合。功能性研究证明PTB可以抑制HepG2.2.15细胞的HBsAg表达量,并呈浓度依赖性。由HPRE引起的HBsAg表达的增加也能被PTB所抑制。实验数据证明PTB通过与HPRE相互作用抑制HBsAg的基因表达。     

Occurrence of HBs antigen and anti-HBs antibodies in analytical laboratory workers in Warsaw

The study aimed at answering the question whether markers of the viral hepatitis, namely HBs antigen and anti-HBs antibodies, are significantly more frequent in the personnel of the analytical laboratories than in blood donors of the City Blood Donation Centre. Together 1,284 persons employed at 88 analytical laboratories were examined. These persons were divided into the groups according to the occupation, age and duration of the employment. HBs antigen was detected with EIA technique in 13 subjects making 1,025% of all examined individuals whereas anti-HBs antibodies were detected with EIP technique in 20 subjects, i.e. 1,560%. Detectability of HBs antigen and anti-HBS antibodies in blood donors was 0.443% and 0.04% respectively. The obtained results indicate significantly more frequent occurrence of both markers in the employees of the analytical laboratories.     

Comparative biophysical studies of hepatitis B antigen, subtypes adw and ayw.

Comparative biophysical and biochemical analyses were performed on purified preparations of hepatitis B antigen (HBs Ag) subtypes adw and ayw, including isoelectric pH evaluations, analysis of the different morphological forms, molecular weight determinations, and analysis of the polypeptides by polyacrylamide gel electrophoresis, Both HBs Ag-positive plasma and purified HBs Ag were analyzed by electrofocusing in a sucrose ampholyte gradient. Four distinct populations of HBs Ag with a pH range of 4.5 plus or minus 0.1 to 5.4 plus or minus 0.1 for unfractionated plasma samples and 3.9 plus or minus 0.05 to 4.9 plus or minus 0.05 for purified samples were detected in both adw and ayw preparations. Electron microscopic studies of each population of purified HBs Ag revealed 19- to 27-nm spheres in each fraction. Purified material labeled with 125I by the chloramine-T method behaved as one major population with an isoelectric pH value of 3.9 plus or minus 0.1. Purified adw preparations revealed a major population with a molecular weight of 3.7 times 10-6 and a second one of 4.6 times 10-6. Purified preparations of ayw contained one population with a molecular weight of 4.6 times 10-6. Polyacrylamide gel electrophoretic analysis of purified HBs Ag revealed nine polypeptides for ayw and seven for adw particles. These studies indicate that purified preparations of HBs Ag are heterogeneous and that distinct differences can be detected between the two subtypes.