Hepatitis B Surface Antigen Titers Research Articles

Newly Licensed Hepatitis B Vaccine

THE RECENTLY licensed subunit hepatitis B vaccine (HEPATAVAX-B) is unique among vaccines in that it is manufactured solely from human plasma obtained from asymptomatic individuals with chronic hepatitis B. Plasma from donors selected for manufacturing this vaccine contains high concentrations of noninfectious hepatitis B surface antigen (HBsAg) particles and lower concentrations of infectious hepatitis B virus (HBV). The ratio of HBsAg to HBV can be as great as 10,000:1.¹From the starting plasma, 22-nm spherical HBsAg particles are separated from HBV by ultracentrifugation. The 22-nm particles are then further purified by digestion with pepsin followed by the addition of 8M urea. Finally, the vaccine is treated with formaldehyde. <h3>Vaccine Plasma Donors</h3> Vaccine plasma donors are selected for their high titers of HBsAg. By federal regulation, they must be asymptomatic and in apparent good health.²They must meet all federal requirements for acceptable plasmapheresis donors, except that their serum

Clinical aspects of delta infection.

The clinical features of delta infection were analysed retrospectively in 191 hepatitis B surface antigen (HBsAg) carriers and 592 cases of acute hepatitis B seen over 11 years in the Swedish town of Malmö (population 250 000). With a few exceptions delta infections occurred exclusively in drug addicts. In the chronic HBsAg-carriers the most common clinical manifestation was an episode of acute hepatitis, which in some individuals became severe with a pronounced rise in serum alanine aminotransferase activity for many months. During the period of delta infection the HBsAg titre was lowered and in three out of 26 cases the patient lost HBsAg altogether and developed hepatitis B surface antibodies (anti-HBs). In one patient the acute hepatitis due to delta infection was fulminant and fatal. In patients with acute hepatitis B the clinical picture did not distinguish between those with and without simultaneous delta infection. The frequency with which acute hepatitis B was succeeded by a chronic carrier state was the same whether or not the patient was infected simultaneously with the delta agent. The discovery of the delta agent has improved understanding of the natural history of chronic hepatitis B infection in drug addicts. Thus, instances of acute hepatitis in a chronic carrier, previously termed hepatitis non-A, non-B, may actually be episodes of delta infection.

Effects of Immunosuppressive Therapy on Viral Markers in Chronic Active Hepatitis B

Hepatitis B virus associated DNA polymerase activity, hepatitis b surface antigen (HBsAg), and serum aspartate aminotransferase were followed in 21 patients with chronic active hepatitis while immunosuppressive therapy (prednisone +/- azathioprine) was being withdrawn. In every case, DNA polymerase activity fell within 6-10 wk of decreasing treatment and became undetectable in 8 patients. This was usually accompanied by a fall in HbsAg titer and a transient rise in serum aspartate aminotransferase activity. Four additional patients with previously untreated HbsAg positive chronic active hepatitis were placed on prednisone for 12 wk. There was a rise in DNA polymerase activity and HBsAg titer with a fall in serum aspartate aminotransferase values during treatment. Upon discontinuing therapy, DNa polymerase activity fell dramatically in all 3 patients who completed their course of prednisone and became undetectable in 1. These findings suggest that immunosuppressive therapy has a potentiating effect on hepatitis B viral replication in patients with chronic active hepatitis.

Seroconversion from hepatitis B e antigen to antibody in chronic type B hepatitis.

Twenty-five patients with chronic type B hepatitis documented by liver biopsy were followed for 1 to 6 years with serial measurements of aminotransferase levels, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and antibody (anti-HBe), and hepatitis B virus DNA polymerase. Initially, all were positive for HBsAg and HBeAg and had elevations in serum aminotransferases. In follow-up, only one lost HBsAg reactivity. In 13, however, elevated aminotransferase levels spontaneously fell to normal and have remained normal. These 13 also had a seroconversion from HBeAg to anti-HBe, and all became negative for serum DNA polymerase. Most had a fall in HBsAg titer. This seroconversion occurred concurrently with or several months before the fall in aminotransferase levels. In contrast, the 12 persons who remained HBeAg positive continued to have elevated aminotransferase levels. All 10 of these patients who were initially positive for DNA polymerase remained positive. These data suggest that many patients with chronic type B hepatitis eventually have a spontaneous remission in clinical and biochemical evidence of active disease, usually heralded or accompanied by the disappearance of HBeAg and DNA polymerase.

Primary hepatocellular carcinoma with hepatitis B antigenemia: effects of chemotherapy.

Eight patients with primary hepatocellular carcinoma (PHC) whose blood was positive for hepatitis B surface antigen (HBsAg) received treatment with cyclophosphamide, adriamycin, and 5-fluorouracil. In 4 cases, a rise in serum HBsAg titer occurred after chemotherapy. In 2 cases involving drug-related leukopenia, the rise in HBsAg titer was marked and associated with a sharp rise in the serum transaminase level (SGOT), up to 1700 in 1 case. Lower-dosage chemotherapy was safely resumed after SGOT had returned to pretreatment levels. No evidence of immunodeficiency after chemotherapy was revealed by in vitro testing of lymphocyte and granulocyte function, percentage of circulating T-cells, and immunoglobulin and complement levels. All 8 cases were negative for e antigen (eAg) and 4 were anti-positive. In 3 of 4 cases, anti-e became negative after chemotherapy, but all remained eAg negative. The negative eAg tests in these cases of PHC suggest they are not highly infectious, in spite of increased HBsAg titers in blood following chemotherapy.

Hepatitis B e antigen in volunteer and paid blood donors.

Sera from 200 volunteer donors and 200 paid blood donors, all positive for hepatitis B surface antigen (HBsAg), were tested for the presence of hepatitis B e antigen (HBeAg).HBeAg was detected in 31 HBsAg-positive paid donors (15%), and in 11 HBsAg-positive volunteer donors (5%) by agar gel diffusion. The presence of HBsAg was associated with higher titers of HBsAg. No significant difference was found in the prevalence of antibody to HBeAg (anti-HBe) in the two donor groups. Rheumatoid factor was not associated with the presence or absence of HBeAg or anti-HBe, indicating that HBeAg is probably not an anti-IgG. These data support the epidemiological evidence that paid blood donors appear to be more likely than volunteer donors to transmit hepatitis B virus infection to recipients of their blood.

Hepatitis A and B virus markers in immune serum globulin

Hepatitis A and B virus markers were investigated in 62 Jots of immune serum globulin (ISG) prepared by four United States manufacturers over the years 1962–1977. All lots were positive for antibody to hepatitis A virus (anti-HAV) in moderately high titer, and there was no apparent change in anti-HAV titer over the 15-yr period. In contrast, only 48% of ISG lots contained detectable antibody to hepatitis B surface antigen (anti-HBs), and titers varied greatly. There was a marked increase around the year 1972 in both the percentage of lots positive (from 29% to 100%) and in the mean anti-HBs titer (from < 1:10 to 1:500) in ISG. All ISG lots were positive for antibody to hepatitis B core antigen (anti-HBc), but titers varied markedly. The titer of anti-HBc appeared to change around the year 1972, in this case falling from a mean of 1:8000 to 1:800.All ISG lots were negative for hepatitis B surface antigen (HBsAg) when tested directly by radioimmunoassay. However, after ultracentrifugation through an acid-sucrose gradient (a technique devised to render HBsAg complexed with anti-HBs detectable), 58% were found to contain small amounts of HBsAg. This HBsAg reactivity was present in 78% of ISG lots prepared before 1972, but in none manufactured since. The elimination of HBsAg from ISG was probably due to the introduction of routine HBsAg screening of plasma donors in 1972. The simultaneous rise in titer of anti-HBs in ISG can also be explained by this screening, since HBsAg contaminating plasma pools would complex with and absorb any free anti-HBs activity, making it undetectable. The presence of HBsAg and variable titers of anti-HBs in ISG preparations may have influenced the outcome of clinical trials of ISG in preventing type B hepatitis.

HBeAg and anti-HBe detection by radioimmunoassay: correlation with vertical transmission of hepatitis B virus in Taiwan.

A sensitive radioimmunoassay technique was used to detect hepatitis B e antigen (HBeAg). A strong correlation was found between HBeAg positivity of the serum of hepatitis B surface antigen (HBsAg) carrier women in Taiwan and the subsequent development of surface antigenemia in their babies. All babies who became chronic HBsAg carriers were born to HBeAg positive women, maternal HBeAg positivity being a better prior indication of chronic antigenemia developing in the baby than the HBsAg titer in the mother's serum.

Liver enzyme concentrations as measure of possible infectivity in chronic asymptomatic carriers of hepatitis B.

Fifty-two British-born blood donors who were chronic carriers of hepatitis B surface antigen (HBsAg) were tested for the presence of hepatitis B e antigen (HBeAg) and antibody to HBeAg by an immunoradiometric assay. The presence of HBeAg was closely associated with a slight rise in serum liver enzyme concentrations, a high HBsAg titre, and male sex. We suggest that the finding of persistently raised serum liver enzyme concentrations in an asymptomatic HBsAg carrier might be useful as a likely indicator of HBeAg and high infectivity.

E-Antigen: a link between immune response and infectivity in hepatitis B?

In a study of the distribution of e-antigen and anti-e in subjects whose blood was positive for hepatitis B surface antigen (HBsAg), patients with acute hepatitis B who were tested during the incubation period were all e-antigen-positive but after the onset of illness e-antigen was detected in only 11%. Persistence, and in some instances reappearance of e-antigen in those who became long-term carriers of HBsAg was associated with high titres of HBsAg. There was a high incidence of e-antigen in those conditions in which cell-mediated immunity may be depressed, including Down's syndrome and chronic renal failure. The majority of HBsAg carries identified as sources of infection were e-antigen-positive. A postive reaction for e-antigen is evidently associated with a defective immune response to hepatitis B virus infection which permits continued replication of virus in liver cells accompanied by high titres of HBsAg, numerous Dane particles and detectable DNA polymerase in the blood with consequently a greater likelihood of transmitting infection. Although it cannot be assumed that anti-e positive carriers of HBsAg are not infective, it may be necessary, in the assessment of passive or active immunization for the control of hepatitis B, to take into account the e-antigen/antibody status of possible sources of infection.

HBsAg-positive chronic liver disease: inhibition of DNA polymerase activity by vidarabine.

Four patients who had chronic liver disease and were positive for hepatitis B surface antigen (HBsAg) were treated with vidarabine, a synthetic purine nucleoside that inhibits DNA polymerase activity in vitro and in vivo. Before treatment all had raised serum DNA polymerase concentrations. Three also had hepatitis B e (HBe) and were shown by electron microscopy to have hepatitis B virus (Dane) particles in their serum. In all patients 10 days' treatment with vidarabine resulted in an immediate loss of DNA polymerase activity. In three patients the activity returned when treatment was stopped. In those three patients Dane particles and HBe antigen persisted during and after treatment; in the fourth patient, who remained negative for DNA polymerase, HBsAg titres fell. Although vidarabine inhibited virus replication, virus particles did not disappear from the blood in these patients, presumably because the particles were cleared only slowly. Similar results with interferon suggest that the virus disappears, and HBsAg titres fall, some weeks after the fall in DNA polymerase activity. Continued treatment may therefore have a sustained effect on viral replication. Whether vidarabine can permanently clear HBsAg and so arrest chronic liver disease remains to be seen, but at the very least it could reduce the spread of infection.

Hepatitis B surface antigen-specific cell-mediated immune responses in human chronic hepatitis B surface antigen carriers.

The presence of hepatitis B surface antigen (HBsAg) and antibody (anti-HBs), hepatitis B e antigen (HBeAg) and antibody (anti-HBe), the nature of T-cell function, and specific cell-mediated immunity to HBsAg were determined and evaluated serially in groups of subjects with chronic HBsAg carrier states and in seronegative controls. The techniques of in vitro lymphocyte transformation, spontaneous rosette formation, radioimmunoassay, reverse passive hemagglutination, passive hemagglutination, rheophoresis, and liver function tests were employed for these studies. For the lymphocyte transformation assay, multiple concentrations of phytohemagglutinin and purified HBsAg were used as stimulants. Cell-mediated immunity to HBsAg was detectable in 50% of the chronic HBsAg carriers (responders) at one or more concentrations of HBsAg. The remaining carriers (nonresponders) and controls failed to manifest HBsAg-specific lymphocyte transformation activity. The profile of the responders was characterized by elevated serum glutamic pyruvic transaminase levels, the presence of anti-HBe, high HBsAg titers, and the conspicuous absence of HBeAg in the serum. The nonresponders were characterized by normal serum glutamic pyruvic transaminase levels, the presence of HBeAg and anti-HBe, and lower HBsAg titers. These observations demonstrate the presence of specific cell-mediated immunity to HBsAg in chronic HBsAg carriers who manifest biochemical evidence of liver disease.

Relationship of subtypes of hepatitis B surface antigen with e antigen and its corresponding antibody.

A total of 3,212 samples of blood from asymptomatic volunteer donors positive for hepatitis B surface antigen (HBsAg) by radioimmunoassay and 860 samples negative for HBsAg were tested for the presence of hepatitis e antigen (HBeAg) and its corresponding antibody (anti-HBe) using a rheophoresis technique. Neither HBeAg nor anti-HBe was detected in any sample negative for HBsAg. Of the HBsAg-positive samples, 237 (7.4%) contained HBeAg and 749 (23.3%) had anti-HBe. HBeAg was significantly (P less than or equal to 0.0005) more likely to be in samples with high titers of HBsAg. Anti-HBe had an equal chance of occurring in samples with low or high titers of HBsAg. Both HBeAg and anti-HBe were found in samples with HBsAg subtypes ad or ay. However, a significant association (P less than or equal to 0.0005) between HBsAg/ay and HBeAg as well as an association between anti-HBe and HBsAg/ad was found.

Hepatitis B Surface Antigen and e Antigen in Pleural Effusion: A Case Report

Hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) were demonstrated in pleural fluid obtained from a 40-year-old dialysis patient who had HBSAg and HBeAg in her serum. The titers of HBsAg and HBeAg were similar in serum and pleural fluid. Examination of the pleural fluid by electron microscopy revealed the presence of intact hepatitis B virus particles. These findings underline the potential hepatitis B virus infectivity of other body fluids besides blood, especially for medical and laboratory personnel.

Transmission of hepatitis B to chimpanzees by hepatitis B surface antigen-positive saliva and semen.

To assess the infectivity of hepatitis B surface antigen (HBsAg)-containing body fluids other than blood, chimpanzees were inoculated intravenously with saliva and semen obtained from HBsAg-positive individuals implicated in non-percutaneous transmission of hepatitis B. Saliva and semen samples were negative for occult blood. The titer of HBsAg in saliva was on the average only 1/3,000 that of the corresponding serum. One chimpanzee, inoculated sequentially with saliva from three individuals, developed HBsAg at 9 weeks and serum glutamic pyruvic transaminase elevation at 13 weeks after injection. HBsAg persisted for 15 weeks. This animal also developed e antigen, anti-core antibody, and anti-surface antibody. Liver biopsies showed acute hepatitis that subsequently resolved. A second chimpanzee, inoculated with HBsAg-positive semen, developed HBsAg and elevated serum glutamic pyruvic transaminase 4 weeks after inoculation and then died suddenly without explanation. HBsAg was positive in two consecutive samples and was confirmed by specific neutralization. Autopsy did not reveal evidence of hepatitis. This study demonstrates that HBsAg-positive saliva and, probably, semen contain infectious virus and suggests that saliva and/or semen may serve as important mechanisms in the transmission of type B hepatitis.

The e antigen and vertical transmission of hepatitis B surface antigen.

The relationship of e antigen (eAg) and its antibody (anti-e) to vertical transmission of hepatitis B surface antigen (HBsAg) from chronic asymptomatic HBsAg carrier women to their children was investigated in Taiwan. Sera from 20 of the 62 women studied were positive for eAg (32%); serum from only one woman was positive for anti-e (2%). A total of 85% of the babies born to eAg positive mothers became HBsAg carriers, while only 31% of the babies became carriers when the mother was eAg negative. Maternal e antigenemia correlated with a high HBsAg titer, and both parameters were equally good predictors of vertical transmission.

Detection, by three techniques, of hepatitis B surface antigen (HBsAg) and determination of HBsAg and anti-HBs titres in patients with chronic liver disease.

The sensitivities of three technqiues used to detect serum hepatitis B surface antigen (HBsAg) were compared in 411 patients with various types of chronic liver disease. Counterimmunoelectrophoresis proved an unreliable test. Two haemagglutination technqiues were slightly less sensitive than radioimmunoassay but were more rapidly performed. Less sensitive techniques were particularly unreliable in active liver disease where HBsAg titres were low. HBsAg was detected in patients with chronic persistent hepatitis, alcoholic liver disease, chronic active liver disease with or without cirrhosis, and primary liver cell carcinoma. Forty-six of the 68 (68%) HBsAg positive subjects were males coming from outside the United Kingdom. The HBsAg titres in 13 subjects with chronic persistent hepatitis were significantly higher (P less than 0-001) than those in 43 subjects with chronic active liver disease. Corticosteroid therapy did not alter the HBsAg titre significantly. None of the 28 HBsAg positive subjects studied serially for up to two years cleared HBsAg from the serum. Anti-HBs was examined by passive haemagglutination and found in 35 subjects, 26 of whom had no evidence of liver disease, 80% came from abroad. Anti-HBs was believed to be of epidemiological rather than of pathological importance.

Induction of Antibody to the “y” Determinant of HBsAg in a Chimpanzee Carrier of HBsAg Subtype “adw”

Antibody to the y determinant of hepatitis B surface antigen (HBsAg) was induced in a chimpanzee chronically infected with hepatitis B virus and circulating HBsAg subtype adw. The chimpanzee was immunized with purified preparations of HBsAg subtypes adw and ayw. Six weeks after immunization, antibody to HBsAg (anti-HBs) specific for the y determinant, appeared. No change occurred in titers of HBsAg or antibody to hepatitis B core antigen (anti-HBc) and "e" antigen remained detectable. The circulating HBsAg subtype adw remained present despite the persistence of anti-y for greater than 8 months.

Association of Dane Particles with e Antigen in the Serum of Asymptomatic Carriers of Hepatitis B Surface Antigen

Asymptomatic carriers of hepatitis B surface antigen (HBsAg) were surveyed for serum e antigen (e Ag) and antibody to e (anti-e) by immunodiffusion. Seventeen serum samples containing e Ag and 17 containing anti-e, but with similar HBsAg titers, were tested for the presence of hepatitis B core antigen (HBcAg) by means of immune adherence hemagglutination method. HBcAg activity was detected in all of the samples containing e Ag, and the titer of HBcAg paralleled that of HBsAg in each serum. In remarkable contrast, no HBcAg activity was found in any of the serum samples containing anti-e. Dane particles were demonstrated in 14 of 17 serum samples with e Ag by electron microscopy, but in none of 17 with anti-e. Although both e Ag and HBcAg were closely associated with each other, the non-identity of e Ag and HBcAg was clearly demonstrated by a two-dimensional immunodiffusion test.

EVIDENCE AGAINST BREAST-FEEDING AS A MECHANISM FOR VERTICAL TRANSMISSION OF HEPATITIS B

A follow-up study of 147 babies born to mothers known to be carriers of hepatitis B surface antigen (HBsAg) revealed no evidence for a relationship between breast-feeding and the subsequent development of antigenæmia in the babies. All babies were tested at three or more months of age, and the mean age at last follow-up was eleven months with a mean of three serum specimens per baby (not counting cord-blood specimens). The frequency of acquisition of HBsAg and anti-HBs was almost identical among breast-fed and non breast-fed infants. The frequency of other variables known to influence the development of antigenæmia among infants of carrier mothers did not vary according to breast-feeding status: mother's HBsAg titre, HBsAg positivity rate in cord-blood specimens, and HBsAg prevalence among siblings. 32 breast-milk specimens from carrier mothers were mothers were all HBsAg negative.