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Application of a screening test for antibody the hepatitis B core antigen.

Hepatitis B core antigen (HBcAg) was prepared from human liver tissue and used in an immunoelectro-osmophoresis screening test to detect antibody to HBcAg (anti-HBc) in patients with evidence of liver disease and in blood donors. With the exception of two immunosuppressed HBsAg carriers, anti-HBc was found in all cases of hepatitis B infection even when HBsAg was detectable only by radioimmunoassay. Non-specific reactions were observed in 'non-B' hepatitis but, in spite of this problem, antib-HBc screening was considered a useful addition to routine tests in the clinical hepatitis laboratory.

Acute Disease

Antibodies to hepatitis B core antigen in hepatitis B surface antigen-positive and -negative chronic hepatitis.

Antibody to hepatitis B core antigen (anti-HBc) is a sensitive indicator of hepatitis B virus (HBV) infection. However, anti-HBc has been found in only a few patients with chronic hepatitis. Therefore, we tested for anti-HBc in 124 sera from 67 patients with histologically proven chronic hepatitis by the indirect fluorescent antibody technique. All patients, except for one with chronic hepatitis who was seropositive for hepatitis B surface antigen (HBs Ag), had anti-HBc that persisted throughout the follow-up period (three months to three years). Of 33 HBs Ag-seronegative patients, anti-HBc was detected in seven patients and persisted for six months to two years. These findings suggest that in this study 21% of patients with chronic hepatitis with undetectable amounts of HBs Ag in the serum had evidence of recent or continued HBV replication.

Adolescent

Differential distribution of hepatitis B surface antigen and hepatitis B core antigen in the liver of hepatitis B patients.

One hundred liver biopsies from 100 patients with clinical presumptive diagnosis of hepatitis were examined by immunofluorescence for the presence of hepatitis B surface antigen (HBSAg) and hepatitis B core antigen (HBcAg). Of the 60 HBsAg-positive livers, 51 were diagnosed as chronic hepatitis on histological grounds, 6 as acute hepatitis, and 3 as "near-normal liver." From the 60 tissue-positive cases, 3 subjects were HBsAg seronegative. HBcAg was detected in 44 livers, all of which also had HBcAg in the localized in the cytoplasm and the membranes of the hepatocytes, and HBcAg in the nuclei and in 4 cases also in the cytoplasm. Predominant HBsAg expression in the cytoplasm was observed in near-normal liver, chronic persistent hepatitis, and cirrhosis with little activity. This correlated with the amount of ground glass hepatocytes in the biopsies. HBcAg and membrane-localized HBsAg were minimal in those conditions. HBcAg was most prevalent in patients with chronic aggressive hepatitis and active cirrhosis treated with immunosuppressive drugs, whereas the amounts of HBsAg and HBcAg in nontreated patients of those two groups and in acute hepatitis with signs of transition to chronicity were almost equal. HBsAg expression in liver cell membranes was most prominent in active forms of chronic hepatitis (chronic aggressive hepatitis and in active cirrhosis) and in acute hepatitis with signs of transition to chronicity. This observation correlated in the presence of HBcAg in the biopsies of those patients. In acute hepatitis both HBsAg and HBcAg were detected rarely and no membrane expression of HBsAg was observed. The over-all results show a significant relationship between the different degrees of accumulation of HBsAg and HBcAg in the liver and the various histological types of hepatitis and further suggest an interplay of both hepatitis B virus and host immune response in the development and pathogenesis of hepatitis B.

Acute Disease

Antibody to hepatitis B core antigen in chronic active hepatitis.

Antibody to hepatitis B core antigen (anti-HBc), which has been assumed to be a more sensitive indicator of hepatitis B virus replication than hepatitis B surface antigen (HBsAg), was detected in the sera of 26 of our 65 patients with HBsAg-negative chronic active hepatitis. Thus despite the absence of HBsAg the liver disease could be the consequence of chronic infection with hepatitis B virus in these patients. They differed, however, from a group of 35 patients with HBsAg-positive hepatitis in being older on average and having less active liver lesions. The two groups could represent either two stages of chronic infection with hepatitis B virus or two types of response to it.

Adolescent

Dane particles-associated hepatitis B core antigen in patients with HBsAg-positive chronic hepatitis.

Dane particles-associated hepatitis B core antigen (HBcAg) was determined by radioimmunoassay in 61 patients with hepatitis B surface antigen (HGsAg)-positive chronic hepatitis. HBc antigenemia was observed in 61% of patients, especially in those with epidemiological risk factors. Patients with chronic active hepatitis as well as those with chronic persistent hepatitis may have HBc antigenemia. The highest levels of HBcAg were observed in male homosexuals. Follow-up determinations indicate the general tendency of HBcAg to decrease or disappear. HBcAg-positive patients with chronic active hepatitis had a poor prognosis, whereas HBcAg-negative patients frequently had a favorable clinical course of the disease (P less than 0.001). The assay of HBcAg in the serum of patients with HBsAg-positive chronic active hepatitis is a useful parameter with both clinical and epidemiological importance.

Adult

Detection of e antigen and antibody: correlations with hapatitis B surface and hepatitis B core antigens, liver disease, and outcome in hepatitis B infections.

Testing for e antigen and antibody (anti-e) was performed by immunodiffusion and counterelectrophoresis in patients with polyarteritis nodosa fulminant hepatitis, and chronic active hepatitis (CAH), in 59 asymptomatic carriers of hepatitis B surface antigen (HBsAg) who underwent liver biopsy, and in 150 carriers followed with sequential SGPT determinations. Counterelectrophoresis was more sensitive that immunodiffusion. Neither e antigen nor anti-e was found in the absence of HBsAg. Among HCsAg-positive patients with polyarteritis nodosa and CAH, e antigen was found in 16 of 18 and 13 of 22, respectively. It was not found in any of 43 patients with fulminant hepatitis, of whom 24 were HBsAg-positive. The e antigen was detected in none of 13 biopsied carriers with normal histology, 4 of 28 with nonspecific changes of 11 of 18 with CAH or chronic persistent hepatitis. Conversely, anti-e was present in 9 of 13 with normal biopsy, 7 of 28 with nonspecific changes, and none of 18 with CAH or chronic persistent hepatitis. The e antigen was found more commonly in nonbiopsied carriers with elevated SGPT, and anti-e in those with normal SGPT. Six carriers whose antigenemia terminated spontaneously had anti-e. The presence of e antigen correlated with a high titer of HBsAg, and with immunofluorescent detection of hepatitis B core antigen in the nuclei of hepatocytes. Conversely, anti-e was associated with significantly lower titers of serum HBsAg (P less than 0.001) and lack of detectable hepatitis B core antigen in the liver.

Alanine Transaminase

Hepatitis B core antigen immune complexes in the liver of hepatitis B patients.

Single liver biopsies from 102 clinically diagnosed hepatitis patients were examined by immunofluorescence for the presence of hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg), complement and immunoglobulin deposition, and for their capacity to fix human complement in vitro. Of the sixty-five HBsAg positive livers, fifty-three were histologically diagnosed as chronic hepatitis, three as acute hepatitis, five as acute hepatitis with signs of transition to chronicity, and four as 'near normal liver'. In the group with chronic hepatitis, HGcAg was observed in thirty-nine livers, all of which also had HBsAg. Thirty-five of these thirty-nine cases also had the ability to fix complement in vitro in the hepatocyte nuclei and/or cytoplasm. Of these thirty-five cases, twenty-nine were positive for immunoglobulin deposition on the nuclei. All of these cases had antibody to HBcAg in the blood, but only five had anti-HBs. The frequency of in vitro complement fixation and immunoglobulin deposition was higher in active forms of the disease, such as chronic aggressive hepatitis and active cirrhosis, than in non-active disease such as chronic persistent hepatitis and mild cirrhosis. By the application of double fluorescent staining techniques, complement fixation was observed in some HBcAg-positive nuclei. In the 'near normal liver' cases there was no intrahepatic accumulation of HBcAg, and despite the presence of anti-HBc in the blood, in vitro complement fixation and immunoglobulin deposition were both absent. The group of three HBsAg ositive 'acute hepatitis with signs of transition to chronicity' cases behaved similarly to those with chronic aggressive hepatitis and had circulating anti-HBc, in vitro complement fixation and immunoglobulin deposition in the hepatocytes. None had circulating anti-HBs. In the group sith HBs-positive acute hepatitis, anti-HGc in the blood was the only other evidence of hepatitis B virus infection.

Antigen-Antibody Complex

Antibody to hepatitis B core antigen in chronic hepatitis and primary biliary cirrhosis: evaluation by a new autologous solid phase radioimmunoassay.

Anti-HBc, the antibody to core antigen of hepatitis B virions, was assayed by a new solid phase sandwich radioimmunoassay inhibition method in the sera of 26 patients with HBsAg-negative chronic active hepatitis (CAH) and 31 patients with primary biliary cirrhosis (PBC). The sensitivity of the method was validated by finding anti-Hb titers averaging greater than 1:1000 in a group of 8 chronically HBsAG-positive individuals, 4 of whom had chronic persistent hepatitis and 4 of whom had no histological or biochemical evidence of liver disease. However, anti-HBc was not detectable in any of the 26 patients with HBsAg-negative CAH. Sera from 2 of the 31 PBC patients contained anti-HBc but only in low titers, a distribution of anti-HBc similar to that found among a comparison group of medical personnel. Anti-HBc testing among PBC patients and control subjects identified a few persons in whom negative tests for HBsAg and anti-HBs failed to identify previous hepatitis B infections. Nevertheless, the uniformly negative tests for anti-HBc among our HBsAg-negative patients with CAH provide additional evidence that this subgroup, typically young-middle age females, seldom derive CAH from hepatitis B infection.

Adolescent

The diagnostical significance of antibodies against hepatitis B core antigen.

The frequency of hepatitis B was examined using three serological parameters: HBsAg, antiHBs and antiHBc. All three substances were detected qualitatively by means of sensitive radioimmunological techniques. Of 1216 patients with acute hepatitis, 55 percent were HBsAg and antiHBc positive on admission to hospital. A further 17.8 percent had no HBsAg but were antiHBc positive and also partly antiHBs positive. These cases can be divided into 2 groups: In one group, in 8.6 percent of patients had a high antiHBc concentration during the acute phase. Similar antiHBc concentrations were seldom found (0.04 percent) in HBsAg negative blood donors. AntiHBs in the patients was at first mainly negative and then appeared during reconvalescence. These cases were considered to be acute hepatitis type B, although HBsAg was absent. In the second group, comprising 9.2 percent of the patients, antiHBc was present in low concentrations and in majority of cases antiHBs had been present from the beginning. The same antibody constellation was found in 3.6 percent of 2341 blood donors. In the group of patients it is supposed that the acute hepatitis present is not of type B and has a different aetiology. The low concentration of antibody is interpreted to be a sign of an earlier HBV-infection.

Acute Disease

Correlation between titer of antibody to hepatitis B core antigen and presence of viral antigens in the liver.

Liver specimens of 31 autopsied cases of liver cirrhosis who had had detectable levels of antibody to hepatitis B core antigen (anti-HBc) inthe serum were stained for hepatitis B core antigen (HBcAg) and hepatitis B surface antigen (HBSAg) by the direct immunofluorescence method. Their premortem serum samples were tested for HBSAg, antibody to HBSAg (anti-HBS) and anti-HBC. Persistent hepatitis B virus (HBV) infection as judged by circulating and/or liver HB antigens was identified in 18 patients, and all of them revealed a high titer of anti-HBC ranging from 2(11) to 2(16) by the immune adherence hemagglutination method. In contrast, anti-HBC titer of the remaining 13 patients without detectable HB antigens was less than 2(9), and the geometric mean titer of anti-HBC of the patients with persistent HBV infection was significantly higher than that of the patients without (13.9+/-1.55 versus 7.23+/-1.30; t test, P less than 0.001). A combination of circulating anti-HBS and hepatic HB antigens was found in one patient, whose serum revealed an anti-HBC titer of 2(12). On the basis of these results, a high titer of anti-HBC in the serum (immune adherence hemagglutination titer of 2(11) or more) seems to be a reliable indicator of persistent HBV infection in the liver.

Antibodies

Acute hepatitis: significance of antibody to hepatitis B core antigen.

The value of testing for core antibody (anti HBc) in acute hepatitis was assessed in 503 patients. All hepatitis B surface antigen (HBsAg) positive patients tested were also anti HBc positive. Of the 110 HBsAg negative, anti HBc positive patients, 32 were found to have surface antibody, indicating previous infection with hepatitis B virus (HBV). Of the remaining 78 patients in whom anti HBc alone was detectable, follow-up specimens were received from 28 and, of these, 21 were then found to be anti HBc negative. Thus in acute hepatitis non-specific transient reactions to core antigen may appear, and the presence of anti HBc alone cannot be considered adequate evidence for a diagnosis of HBV infection.

Acute Disease

Antibody to hepatitis-B core antigen in patients with primary hepatic carcinoma.

Antibody to hepatitis-B core antigen (anti-HBc) was assayed in the serum of patients with primary hepatic carcinoma (P.H.C.) and controls from Hong Kong, West Africa, and the United States. In each region the prevalence of anti-HBc was higher in P.H.C. patients than in controls, ranging from 70 to 95% in the patients and from 20 to 68% in the controls from Asia and Africa; 24% of P.H.C. patients and 4% of controls from the U.S. had anti-HBc. These data support the hypothesis that chronic infection with hepatitis-B virus is aetiologically related to P.H.C., especially in Asia and Africa, although other factors must also be involved.

Antibodies, Viral

Radioimmunoassay and some properties of human antibodies to hepatitis B core antigen.

A solid-phase radioimmunoassay for antibodies to hepatitis B core antigen (anti-HBc) is described. Polystyrene beads coated with anti-HBc, hepatitis B core antigen prepared from pooled sera of humans infected with hepatitis B virus (HBV) and 125I-labelled anti-HBc were used for the test. Distinct patterns of development and changes of anti-HBc and their immunologic properties are all related to variations of other markers specific for HBV infections. Knowledge concerning the detailed features of the immune response to hepatitis B core antigen may provide deeper insight into the pathogenesis of HBV infections.

Antibodies, Viral

Antibody to hepatitis B core antigen in patients with hepatocellular carcinoma.

Hepatitis B surface antigen (HBsAg), anti-HBs, and anti-HB core (HBc) were measured in 124 patients with hepatocellular carcinoma (HCC) in comparison with 299 control subjects of comparable ages, and in 48 cases of chronic hepatitis and 52 cases of hepatic cirrhosis. It was found that 72.6% of the HCC patients were positive for anti-HBc, and 80.6% were positive for at least one test, whereas in the control, anti-HBc was positive in 30.1% and 34.1% were positive for at least one test, the differences between the two groups being significant (P less than 0.01). The frequencies of positive tests for HBsAg and anti-HBc were the highest in HCC followed in decreasing order by cirrhosis, chronic hepatitis and the control group. A possible role of HB virus infection in hepatocellular carcinoma is discussed in relation to other factors.

Adult

Complement fixing hepatitis B core antigen immune complexes in the liver of patients with HBs antigen positive chronic disease.

One hundred and fifty-two biopsies from serologically HBsAg positive and negative patients with liver disease were studied in immunofluorescence: for the presence of the surface (HBs) and the core (HBc) antigenic determinants foeterminants of the hepatitis B virus, of immunoglobulins and complement (C) deposits, and for the capacity to fix human C. Circumstantial evidence is presented suggesting that HBc immune-complexes are a relevant feature in the establishment and progression of chronic HBSAg liver disease. C fixation by liver cells was shown in all HBC positive patients with chronic hepatitis; an active form was present in every case, except two with a persistent hepatitis, an inverse ratio of HBc to C binding fluorescence being noted between active chronic hepatitis and cirrhotic patients. HBc without C fixation was observed in only three patients in the incubation phase of infectious hepatitis. IgG deposits were often found in HBc containing, C fixing nuclei. No C binding or IgG deposits were observed in acute self-limited type B hepatitis, in serologically positive patients with normal liver or minimal histological lesions, with and without HBs cytoplasmic fluorescence in their biopsy, or in serologically negative individuals.

Adolescent

Fatal Hepatitis B Reactivation in Absence of Antibody to Hepatitis B Core Antigen in a Lymphoma Patient.

Reactivations of hepatitis B virus (HBV) infection in severely immunocompromised patients with serological profiles of past hepatitis B are non-exceptional and potentially severe or fatal events. The preventive and pre-emptive detection of this serological status is compromised in the absence of antibodies to hepatitis B core antigen (anti-HBc), observed in a very small number of cases of HBV infection. Here, we describe the case of a patient with a serological profile indicating a HBV vaccination although the patient did not report previous vaccination, following lymphoma and chemotherapy including rituximab. This patient presented HBV reactivation with appearance of hepatitis B surface antigen (HBsAg) but without appearance of anti-HBc, suggesting the absence of detectable anti-HBc in this patient at the stage of past infection. Next-generation sequencing provided the complete HBV genome, showing the presence of a mutation in HBsAg associated with immune escape but without the detection of mutations in the core gene previously described as significantly associated with the absence of anti-HBc. Nonetheless, W28* pre-core mutation was found, which was reported to enhance replication capacity in case of weak immune responses and suspected to promote HBV reactivation in association with immune escape mutations. Overall, this case highlights that negativity of anti-HBc cannot definitely rule out past HBV infection and the risk of HBV reactivation in immunocompromised patients in the context of treatment of hemopathies, and that it is difficult to assess such a risk and to prevent or monitor HBV reactivation in patients with past HBV infection but no detectable anti-HBc.

Female

Immunochemical and morphological studies of hepatitis B core antigen isolated from the nuclei of hepatocytes.

Immunochemical and morphological properties of hepatitis B core antigen (HBc Ag) were studied in intranuclear particles isolated from human liver. Immunochemical integrity of the purified particles was indicated in the production by guinea pigs of antibody to HBc Ag (anti-HBc) that was immunochemically identical to human anti-HBc. The HBc Ag particles were 27-30 nm in diameter, displayed apparent icosahedral symmetry, and consisted of distinct subunits. The susceptibility of HBc Ag particles to proteolytic and glycolytic enzymes indicated the presence of proteins and glycoproteins. The structural integrity of core particles depended on disulfide, hydrophobic, and hydrogen bonds, and immunological activity relied on intact sulfhydryl groups. Agents active against lipids did not affect immunological reactivity or core structure, as seen by electron microscopy.

Cell Nucleus

Persistence of serum antibody to hepatitis B core antigen.

The purpose of the present study was to measure the amount of antibody to hepatitis B core antigen (anti-HBc) in different populations by the immunoelectroosmophoresis method. High titers of anti-HBc, up to 1/4,096, were found in the acute stage of hepatitis B virus infections and in the chronic carrier state of hepatitis B surface antigen. In cases of acute hepatitis the anti-HBc titers gradually declined to low levels but persisted for the observation time of 5 to 6 years. Individuals positive for antibodies to hepatitis B surface and core antigens selected from a Swedish "normal" population showed still lower anti-HBc titers, indicating that the hepatitis B infection had occurred earlier. The anti-HBc titers in sera drawn at intervals of 4 years from a group of hemophilia patients may indicate previous infection with replicating hepatitis B virus rather than immunization with noninfectious hepatitis B core antigen material.

Antibodies, Viral