Frequency and Persistence of Serologic Markers Following Three Different Manifestations of Hepatitis B Virus Infection

Abstract. 30 asymptomatic chronic carriers of hepatitis B surface antigen (HB_sAg), 6 asymptomatic blood donors transiently infected with hepatitis B virus, and 38 patients with acute hepatitis B were tested for HB_sAg, anti-HB_s, HB_eAg, anti-HB_e and anti-HB_c.
Comparison of these results revealed significant variation in the frequency of HB_eAg which was present in 1 (3.3%) carrier, 2 (33.3%) of the transiently positive donors, and in 24 (63.2%) of the patients with acute hepatitis. Anti-HB_e was found in 28 (93.3%) of the carriers, 4 (66.6%) of the transiently positive donors, and in 8 (21%) of the patients. Variation was also seen in the strength of anti-HB_c, with only the chronic carriers having titres which were consistently high (above 1,000). Retesting the two groups of donors after a period of approximately 2 years showed no change in the serologic status of the chronic carriers, while amongst the transient HB_sAg positives the 2 HB_eAg reactives had seroconverted, 1 of the anti-HB_e positives had become non-reactive, and 2 of the 6 had developed anti-HB_s. 6 of the patients with acute hepatitis B were serologically reexamined during convalescence and showed results similar to those seen in the transiently HB_sAg-positive donors, with clearance of HB_sAg in all, seroconversion from HB_eAg to anti-HB_e in 4, and the production of anti-HB_s in 4.     

High Prevalence of e Antigen among Healthy Blood Donors Carrying Hepatitis B Surface Antigen in Japan

Abstract. Hepatitis B surface antigen (HB_sAg) was detected by an immune adherence haemagglutination method in the serum samples of 292 voluntary, apparently healthy blood donors at four regional blood centres in Japan. Their serum samples were concentrated 3-fold and tested for e antigen (e Ag) and antibody to e (anti-e) by immunodiffusion. The e Ag was found in 41 samples (14.0%) and anti-e in 57 (18.6%). When 100 randomly selected serum samples containing HB_sAg were tested as they were (unconcentrated), and at 3- and 5-fold concentrations, e Ag was detected in 3,16 and 27, respectively, and anti-e in 10,21 and 26. Subtypes of HB_sAg were similar in carriers with e Ag and with anti-e. There is a high prevalence of e Ag in healthy individuals in Japan. There are also high rates of vertical transmission of hepatitis B virus from mothers to children, as well as a high incidence in the past of post-transfusion hepatitis. This is a further evidence that e antigen is a marker for the infectivity of hepatitis B virus in carriers.     

Subtypes of Hepatitis B Surface Antigen in Native-Born and Immigrant Israelis

Abstract. A survey of HB_sAg among Jerusalem blood donors revealed a prevalence of 0.89% and a predominance of subtype uy (85%) compared with ad (15%). A simultaneous survey of patients with HB_sAg-positive viral hepatitis revealed a similar predominance of a y (85%) compared with ad (1 5%). The uniform distribution of the dominant ay subtype among both carriers and patients, representing a diversity of ethnic and national origins, supports the premise that ad and ay subtypes are preferentially correlated with regional epidemiologic, rather than host or disease-related factors.     

Results of a Multicentre Clinical Trial of the Solid-Phase Enzyme Immunoassay for Hepatitis B Surface Antigen

Abstract. The results obtained in a multicentre clinical trial of an enzyme immunoassay (EIA) method for hepatitis B surface antigen (HB_sAg) (65,451 sera tested) have demonstrated that this new test has a significantly higher sensitivity than a reversed haemagglutination test (rHA). In a part of the trial, EIA was also compared with radioimmunoassay (RIA). Only a small number of discrepant results was obtained with these two tests, indicating similar sensitivities. No definite conclusion about a difference in sensitivities could be drawn from these results. Although the specificity of the EIA screening test is lower than that of rHA and RIA, the mean percentage of false positives was 2.2% of the total number of donor samples screened. Presumptive positives in EIA were subjected to a confirmatory test based on neutralization with human antibodies to HB_sAg. After elimination of false positives in EIA screening, there was excellent agreement between EIA and RIA results.     

ABO Blood Groups and HBsAG Subtypes and Titers in Healthy Carriers

Abstract. The investigation covered 457 'healthy' HB_sAg carriers detected following a screening by CEP. HB_sAg in the sera of these subjects were subtyped by ID and titrated by CEP. The results obtained were analyzed according to the blood group of the cases investigated. Differences were found between the distribution of blood groups among the normal unselected population and among 'healthy' carriers, where the incidence of AB subjects was much higher. The difference was more marked for the carriers of HB_sAg/ad.     

A Pilot Study of Hepatitis B Viral Markers in Volunteer Blood Donors in Kenya, East Africa

Abstract. The occurrence of three major markers of hepatitis B viral (HBV) infection—the surface antigen (Hb_sAg), its specific antibody (HB_sAb), and antibody to the core antigen (HB_cAb)—is reported on 101 Kenyan volunteer blood donors which are tested by highly sensitive techniques.
The surface antigen carrier rate was 14% by radioimmunoassay as opposed to 0.1–1% in Northern Europe and in the USA. The prevalence of the core aad surface antibodies is 53 and 57%, respectively. The core antibody as the only indicator of HBV infection occurred in 3% of Kenyan donors as compared to 0.4 in Caucasian volunteers. The overall infectious pool of HBV in Kenyan donors is 17%. Another application of the core and surface antibody screening of the blood donors has been reported recently in relation to the control of non-A, non-B posttransfusion hepatitis. The coexistence of the two antibodies is observed in 41% of the Kenyan donors after excluding the infectious pool. Exclusion of such a large proportion of donors to control non-A, non-B posttransfusion hepatitis is impractical in Kenya though it can be considered in Europe and in the USA where their prevalence rate is 1–4%. It is relevant to observe, however, that in spite of such high transmission rate of the virus in the community in Kenya, 25% of the adult population have been spared. As the immune status in relation to the HBV of the adult recipients of the blood is likely to be similar to that of the donors, a quarter are susceptible to the virus infection. The susceptible group is of course larger still amongst the paediatric recipients.     

Guillain Barré Syndrome during the Pre-icteric Phase of Acute Type B Viral Hepatitis

Summary: A 54-year-old man presented with the Guillain Barré syndrome (GBS) during the pre-icteric phase of acute type B viral hepatitis. This neurological syndrome has been infrequently described in the clinical course of viral hepatitis but only once previously during the prodromal period. Deposition of circulating immune complexes of the HB_sAg have been implicated in the pathogenesis of arthritis, nephritis and poly-arteritis associated with type B viral hepatitis. Although the relationship of GBS to type B viral hepatitis is uncertain, a similar immunopathological mechanism may be involved.     

Follow-up Studies on Liver Disease Associated with HBS Ag Carriers

Summary: Follow-up studies on liver disease associated with HB_sAg carriers. T. D. Bolin, A. E. Davis and A. G. Liddelow, Aust. N.Z. J. Med., 1976, 6, pp. 539–542.
Eight male subjects who were initially studied in 1972 with liver biopsy because of HB_sAg carrier status were re-studied two years later with liver biopsy, clinical examination and standard liver function tests. Three of the eight subjects remained antigen positive and had continuing liver disease, this being either chronic active hepatitis or chronic persistent hepatitis. Two subjects became HB_s Ag negative and their liver biopsies returned to normal. One subject became HB_s Ag negative but his biopsy disclosed chronic active hepatitis with cirrhosis in the presence of normal liver function tests. While persistence of the antigenaemia is associated with persisting liver disease, the converse is not true in that the disappearance of the antigen does not necessarily imply an improvement in liver disease. Liver biopsy remains the only reliable means of assessing liver disease as biochemical tests of liver function and the clinical finclings may be of little value.     

Subtypes of HBsAg in Eastern and South-Eastern Europe

Abstract. A total of 481 serum samples was collected from asymptomatic carriers of H B,Ag as well as some patients with acute and chronic hepatitis, type B. The sera were obtained mainly from blood banks in the following countries: Poland, Hungary, Romania, Bulgaria, Yugoslavia, and the European part of the Soviet Union. Asymptomatic carriers of HB_sAg from Poland and Hungary showed a preponderance of adw subtype over uyw subtype (Poland -80.7% adw , Hungary -72.2% adw). Carriers from the remaining countries showed the reverse situation: Bulgaria -71.5% ayw , Yugoslavia -79.0% ayw , Romania -82.8% ayw , Soviet Union -87.5% ayw. No individuals were found among these indigenous populations to have the adr or ayr subtypes but two carriers appeared to have the unusual subtype adyw.     

Autoimmune Factors in African Cirrhosis

Sera were tested for autoantibodies in 98 patients with cryptogenic cirrhosis i n Uganda and results correlated with serological tests for hepatitis B surface antigen (HB_s Ag) and antibody to HB_s Ag (HB_s Ab). Smooth muscle antibodies (SMA) were detected in 23 (24%) of the patients but there was no difference in th e incidence of SMA between HB_s Ag-positive and negative cases. Antinuclear antibodies (ANA) were detected in five cases; mitochrondrial, gastric and thyroid antibodies were not found in any patient. Unlike other geographical locations autoimmune mechanisms appear to play little part in the progression of chronic liver disease in Uganda Africans. Hepatitis B surface antigen was present in 36 (37%) and HB_s Ab in 47 (48%) of the patients. Although evidence for past exposure to hepatitis B virus (as shown b y detection of HB_s Ab) wa s present in at least 30 out of the 62 HB_s Ag-negative cases, there was no greater incidence of autoantibodies in HB_s Ag-negative patients with or without HB_s Ab. Persistent infection with hepatitis B virus an d continuing liver damage may be an important factor bu t these results do not favor a role for the virus in causing chronic liver disease by triggering off an autoimmune reaction.     

Low Level Hepatitis B Viremia Detected by Polymerase Chain Reaction Accompanies the Absence of HBe Antigenemia and Hepatitis in Hepatitis B Virus Carriers

Objectives: Because outcome of antiviral treatment in patients with chronic hepatitis (CH) B is difficult to predict, we compared the severity of hepatitis with serum hepatitis B virus (HBV) DNA concentration. Methods: We studied 40 HBV carriers with distinct stages of chronic infection, 32 HBe antigen (HBeAg) -negative or low-grade positive carriers whose HBV strains did not contain a point mutation at nucleotide 1896, 37 HbeAg-negative carriers with or without hepatitis, and 51 HBeAg-positive CH patients treated with interferon. Serum HBV DNA concentration was measured by the end-point dilution method using a polymerase chain reaction (PCR). The point mutation at nucleotide 1896 was detected by restriction fragment length polymorphism with PCR. Results: Among the stages of chronic HBV infection, the serum HBV DNA concentration was lowest (10^{0.67 ± 0.71} copies/μl) in HbeAg-negative asymptomatic carriers. A low-level viremia (10^{2.10 ± 1.45} copies/μl) of HBV strains without the mutation at nucleotide 1896 was associated with an HBeAg-negative state. In HBeAg-negative carriers, the serum HBV DNA concentration in those without hepatitis was significantly lower than in those with hepatitis (10^{1.00 ± 0.89} vs 10^{3.31 ± 1.25} copies/μl, p < 0.0001); 20 of 21 asymptomatic carriers had an HBV DNA concentration below 10² copies/μl. Patients with serum HBV DNA concentrations below 10¹ copies/μl at the end of interferon treatment maintained normal serum alanine aminotransferase concentrations. Conclusions: A serum HBV DNA concentration below 10¹ copies/μl is an important goal for successful treatment of CH-B. PCR is necessary to assess such low-level viremias.     

Recent Advances in our Understanding of Hepatitis B Infections

Abstract. The development of highly sensitive hemagglutination (HA) assays for detection of hepatitis B antibody has made possible observation of primary anamnestic antibody responses in most individuals exposed to hepatitis B virus and the development and evaluation of a hyperimmune γ-globulin preparation.
The hemagglutination inhibition assay, while more sensitive than previous techniques for the detection of hepatitis B antigen (HB-ag) carriers, does not presently appear suitable for use in routine screening of blood donors. However, use of this assay to subtype the antigenic specificities of HB-ag makes it a valuable identity test for ultrasensitive assays.
Prospective serologic follow-up of children newly admitted to an institution for the mentally retarded and studies on HB-ag carriers and their families (by hemagglutination inhibition and HA techniques) have revealed that the ratio of infection to clinical disease in hepatitis B infections is greater than 28:1.     

Hepatitis B surface antigen in blood donors: further observations.

A survey of 128,000 volunteer blood donors from the Greater New York metropolitan area revealed that first-time male donors were positive for hepatitis B surface antigen (HBsAg) 2.5 times more frequently than were first-time female donors; Negroes and Mongols were positive four to 20 times more frequently than Caucasians. The ratio of ad to ay seemed to be higher in non-Caucasian antigen carriers than in Caucasian carriers. Among both Caucasians and non-Caucasians the rate of positivity declined after the age of 50. An excess prevalence of HBsAg was observed in donors with the lowest level of education and in those with the highest level. HBsAg was detectable nine times less frequently among repeat donors than among first-time donors (0.2 vs.1.90 per 1,000). Detection of HBsAg was unrelated to ABO-Rh blood groups. Several mechanisms for these wide variations of antigen detection are possible.     

Hepatitis B virus markers in the population of Dar es Salaam, Tanzania

A total of 542 serum samples from healthy adults (medical students and medical staff, blood donors and pregnant women) residing in or near the city of Dar es Salaam, Tanzania were examined for markers of hepatitis B virus (HBV) infection. Of these samples, 95 (17.5%) were not found to contain any HBV marker when examined by enzyme-linked immunoassay for hepatitis B surface antigen (HBsAg), antibody to hepatitis B surface antigen (anti-HBs) and antibody to hepatitis B core antigen (anti-HBc). HBsAg was demonstrated in 52 (9.6%) samples of which 7 (13.5%) were positive for hepatitis Be antigen (HBeAg) and 17 (32.7%) were positive for anti-HBc IgM. None of 9 HBsAg positive pregnant women were carriers of HBeAg. These results show that hepatitis B infection is very common in this country. The relatively low prevalence of HBeAg among HBsAg carriers may indicate that transmission of hepatitis B at birth is not of major importance.     

Preparation and Properties of a Therapeutic Factor IX Concentrate

Abstract. A method for the large-scale preparation of a coagulation factor IX concentrate from human plasma is described. The method includes absorption of the coagulation factors from cryosupernatant plasma onto DEAE-Sephadex, extensive washing of the gel and elution of the coagulation factors with 0.5 M phosphate buffer at 6.85, followed by desalting of the eluate in a column of Sephadex G-25, then by lyophilization, dissolving and sterile filtration, and finally by freeze-drying of the final product. Experiments performed with HB,Ag-positive plasma demonstrated a decrease in the HB_sAg content by a factor of 10^-5 during the process. The product is inactive in Na-PTT assay.
The process yields an about 100-fold purified factor TX concentrate containing also factors 11, VII, and X, but in relatively smaller amounts. The average yield relative to factor IX is 60%. The batch size has been from 16 to 150 litres of plasma and about 300 batches of the concentrate have been prepared.
About 5,100 bottles (about 4.0 times 10⁶ U of factor IX) of the concentrate have been used in the treatment of patients with haemophilia B. The clinical effect has always been good and the in viro response to factor IX was 1.15±0.30%/U/kg body weight. Two cases of HB_sAg-negative hepatitis that may have been caused by the concentrate, were detected. No thrombotic complications were found.     

Follow-Up of Blood Donors Positive for Hepatitis B Surface Antigen

From 1973 to 1977 in Amsterdam the incidence of hepatitis B surface antigen (HBsAg) in blood donations from new donors was 0.224 and from known donors 0.034%. 65 donors, previously found positive for HBsAg, were re-examined. Persistence of HBsAg in new donors (28 of 31) occurred significantly (p less than 0.0005) more often than in known donors (15 of 34). All carriers were classified into HBeAg (21%) or anti-HBe (79%) by a sensitive Elisa technique. Abnormal liver function tests (LFTs) were observed in 30% of the carriers and were significantly (p less than 0.005) more often found in HBeAg than in anti-HBe-positive carriers. When the LFTs remained abnormal, in almost all (8 of 9) carriers moderate to severe histological liver disease was diagnosed.     

e-Antigen and Anti-e in Two Categories of Chronic Carriers of Hepatitis B Surface Antigen

Abstract. Screening for e antigen and anti- e was performed in 517 HBs antigen chronic carriers, 329 blood donors and 188 hemodialyzed patients, e antigen was detected in 35 blood donors (10.6%): 88% had a high titre of HBs Ag, and 80% had a disturbed liver function. No difference was noted regarding sex and age of the carrier. Anti- e was detected in 33.7% of the blood donors. A significant difference (p < 0.01) was noted between males (29%) and females (46.6%). Anti- e was found more prevalent in young people between 21 and 34 than in older people (p < 0.05). Anti- e was rarely found when the titre of HBs Ag was low or high. 88% of the anti- e carriers had an intermediate titre from 1/1 to 1/32 by CEP.
Both e Ag and anti- e were more prevalent in ad subtype than in ay subtype (p < 0.02 for both). Inside the ad subtype, anti- e is less frequent in adr (6%) than in adw ₂ (42%) and in adw ₄ (57%); p < 0.01. The difference between the prevalence of e Ag in adr (31%) and in adw ₂ (17%) is not significant but the healthy carriers with e antigen are more numerous in adr than in adw ₂ subtype.
e antigen was detected in 53.7% of HBs Ag-hemodialyzed patients and anti- e in 3.7% of these patients. These results are neither correlated with the liver function nor with the state of chronic carrier (27% of e Ag in transient HBs Ag carrier).     

Hepatitis B viral DNA in liver and serum of asymptomatic carriers.

Cloned DNA probes were used to test for hepatitis B virus (HBV) DNA in the liver and serum of 14 asymptomatic hepatitis B surface antigen (HBsAg) carriers and two former carriers. The results were compared with serological markers of HBV infection and liver histopathology. Three groups of carriers were distinguished. In group I, HBV DNA was present in both liver and serum. Hepatitis B e antigen (HBeAg), a marker of viral replication, was uniformly positive in serum. In one individual in this group, viral DNA was also integrated into liver genomic DNA. In group II, lower levels of nonintegrated HBV DNA were detected in the liver, but no HBV DNA was found in the serum. HBeAg was negative and with one exception there were antibodies to HBeAg. Integrated viral DNA was present in each case. In group III, there was no detectable nonintegrated viral DNA in serum or liver, but one individual had integrated sequences. All carriers lacked antibodies to HBsAg and had antibodies to hepatitis B core antigen and demonstrated nonspecific histological abnormalities in the liver. These findings indicate significant quantitative and qualitative differences among asymptomatic HBsAg carriers and suggest that their infectivity may be highly variable.     

Hepatitis B virus DNA in hepatitis B surface antigen-positive blood donors: relation to the hepatitis B e system and outcome in recipients

The presence of hepatitis B virus (HBV) DNA was investigated in 26 hepatitis B surface antigen-positive blood donors. Three donors (12%) were concordantly positive for HBV DNA and hepatitis B e antigen (HBeAg) and had IgM antibody to hepatitis B core antigen (anti-HBc). Two donors (8%) had HBV DNA without HBeAg; both were positive for antibody to HBeAg and lacked IgM anti-HBc. Twenty-one HBV DNA-negative donors had antibody to HBeAg, and all were negative for HBeAg and IgM anti-HBc. Blood units from 16 donors were transfused. A sufficient serological and clinical follow-up was available for 10 HBV-susceptible recipients. Three recipients of HBV DNA-positive blood units were infected irrespective of HBeAg status or presence of IgM anti-HBc. Six (86%) of seven recipients of HBV DNA-negative blood units developed HBV infection. Thus all hepatitis B surface antigen-positive blood donors should still be considered infectious irrespective of status with regard to HBeAg, HBV DNA, and IgM anti-HBc.     

Current topics in hepatitis B

Over two billion people around the world have been infected with hepatitis B virus, of whom over 350 million are chronic carriers. Some 25% of carriers develop progressive liver disease. The annual mortality from hepatitis B infection and its sequelae is 1-2 million people worldwide.The following current topics are reviewed: immunization strategies against hepatitis B and the kinetics and antibody response; the controversy on screening blood donors for anti-core antibodies; mutations of hepatitis B surface antigen, including evidence that not all such mutants are detectable by current laboratory tests and, finally, the introduction of second generation nucleoside analogues for treatment of chronic hepatitis B infection, including treatment of patients with decompensated liver disease and liver transplantation.