HBe Antigenemia Research Articles

Diagnostic Utility of Pre-Genomic Hepatitis B RNA in the Evaluation of HBV/HIV Coinfection.

Newer biomarkers of Hepatitis B virus (HBV) infection and treatment response have not been well-characterized in individuals with HBV/HIV coinfection. Pre-genomic RNA (pgRNA) and quantitative HBsAg (qHBsAg) were used to evaluate the associations with baseline characteristics. Participants included two separate groups - 236 with HBV/HIV coinfection enrolled in a cross-sectional cohort in Ghana and 47 from an HBV nucleoside/nucleotide treatment trial comparing tenofovir to adefovir in the United States. In both cohorts, HBe antigenemia was highly associated with pgRNA and HBV DNA levels. In the treatment cohort, pre-treatment pgRNA serum concentration was 7.0 log10 U/mL, and mean qHBsAg was 201,297 IU/mL. The observed treatment-associated decrease in pgRNA was consistent with a biphasic decline curve that reached second-phase kinetics following treatment week 12. Changes from baseline were significantly correlated with changes in serum ALT (r = - 0.518; P = 0.023) but not with changes in HBV DNA (r = 0.132, P = NS). qHBsAg also correlated with ALT change (r = - 0.488, P = 0.034). pgRNA and qHBsAg represent newer biomarkers of HBV replication that may help monitor response and treatment outcomes. HBV pgRNA is highly associated with both HBeAg and ALT and may predict both active replication from the closed circular DNA (cccDNA) template as well as hepatic injury.

The HLA-G 14-base pair deletion allele and the deletion/deletion genotype are associated with persistent HBe antigenemia in chronic hepatis B infection

Background and aims: HLA-G has well-recognized immunomodulatory properties, and this molecule is frequently expressed in the livers of hepatitis B virus (HBV)-infected patients. Because the HLA-G 14 bp-insertion/deletion polymorphism (rs371194629) has been associated with the magnitude of HLA-G expression, we evaluated this polymorphism in the recognized evolutionary forms of chronic HBV infection. Methods: We studied 196 chronic HBV-infected patients (118 HBeAg-negative chronic hepatitis, 53 HBeAg-positive chronic hepatitis and 25 inactive carriers exhibiting low levels of serum HBVDNA and persistently normal ALT levels), and 202 healthy individuals. Chronic hepatitis HLA-G typing was performed using PCR-amplified DNA hybridized with specific primers. Results: The frequencies of the insertion/deletion alleles and genotypes were very similar in patients and controls. After patient stratification according to the evolutionary form of the chronic HBV infection, the frequencies of the deletion allele (P=0.0460; OR=1.26; 95%CI=1.01–1.45) and of the deletion/deletion genotype (P=0.0356; OR=2.08; 95%CI=1.05–4.09) were overrepresented in HBeAg-positive patients when compared to HBeAg-negative patients. No differences were observed when HBV inactive carriers were compared to HBeAg-negative chronic hepatitis patients. Conclusions: Because the 14-bp deletion allele has been associated with increased HLA-G production and because HLA-G may down regulate the cytotoxic activity of TCD8 and NK cells, patients exhibiting the 14-bp deletion allele at single or double doses are at increased risk for developing chronic forms of HBV associated with persistent viremia and worse prognoses.

Epidemiologic trends in HBV infections at a reference centre in Turkey: an 11-year retrospective analysis

Objective. To identify changes in hepatitis B epidemiology after the implementation of the nationwide vaccination program in Turkey, hepatitis B virus (HBV) and related tests performed over a period of 11 years (2000-2010) at a reference centre were retrospectively overviewed and statistically analysed for trends.Results. Assay results for Hepatitis B surface antigen (HBsAg) and e antigen (HBeAg), Anti-HBs, Anti-HBe, Anti-HBc immunoglobulins and HBV DNA as well as aspartate aminotransferase (AST), alanine aminotransferase (ALT), gama-glutamyl transpeptidase (GGT) and alkaline phosphatase (AP) levels, obtained via standardized commercial assays were included in the analysis. Overall, a stable anti-HBs incidence (43.6%) and male predominance in infected individuals were noted. Total Anti-HBc was detected in 43.3% of the Anti-HBs reactive population, demonstrating that the immunity against HBV has still been acquired through virus exposure. An intermediate HBsAg seroprevalence of 6.0% was observed with a significant decrease from 12.3% to 5.0% from 2000 to 2010. Anti-HBe positive infections were more frequent than those with HBe antigenemia (77.1% vs. 18.5%) with a notable increase from 2000 to 2003. HBV DNA was detected in 23.6-25.6% with serological markers of viral replication and was more prevalent in HBeAg positive individuals in parallel with AST, ALT and GGT levels. Evidence for horizontal transfer as the major transmission route was revealed with a reduction of childhood HBV infections, attributable to the ongoing vaccination efforts.

High HBV Viral Loads in HIV-Infected Pregnant Women at a Tertiary Hospital, South Africa

High HBV Viral Loads in HIV-Infected Pregnant Women at a Tertiary Hospital, South Africa

Impact of Universal Hepatitis B Vaccination on the Prevalence of HBs and HBe Antigenemia Among Pregnant Women in Taiwan

Impact of Universal Hepatitis B Vaccination on the Prevalence of HBs and HBe Antigenemia Among Pregnant Women in Taiwan

Epidemiology, diagnosis and treatment of chronic hepatitis B in HIV-infected patients (EPIB 2005 STUDY)

To describe the characteristics of hepatitis B (HBV) infection in HIV-infected patients and the impact of anti-HBV treatments. All the patients with past or present chronic HBV infection seen in October 2005 in 17 French hospitals were included. Data were retrospectively collected from their first visit in a time-dependent manner, through a detailed standardized questionnaire. Among 477 HBV-infected patients, 261 (55%) were co-infected with HIV. The HBV-HIV co-infected patients underwent fewer serological, virological and histological evaluations. Initial positive HBe antigenemia (HBe Ag) was more frequent in these patients (57.9 versus 28.6%; P < 10), as was cirrhosis on the initial liver biopsy (17.9 versus 7.6%; P = 0.05). Throughout the mean 5-year follow-up, HBe Ag loss was less frequent (P = 0.04), as was HBe seroconversion (incidence rate 2.6 versus 10/100 patient-years; P < 10). HBe Ag loss was associated with fibrosis improvement (METAVIR score -0.5 +/- 0.4 versus +0.2 +/- 0.6 if persistent positive HBe Ag, P = 0.01). In co-infected patients on tenofovir, adefovir or interferon, HBe seroconversions were seen in patients on combined HBV treatment, the use of which is increasing (58% in 2005). Nevertheless, no significant difference in virological, immunological or biochemical evolution was observed between these different treatments. In HBV-HIV co-infected patients, the assessment of HBV infection still needs to be improved, the HBV wild-type remains predominant, and HBe Ag loss is rare and associated with a better histological evolution. There is insufficient evidence of the superiority of combined HBV treatment, and this still needs be demonstrated in long term studies.

Polyarteritis Nodosa and Extrahepatic Manifestations of HBV Infection: The Case Against Autoimmune Intervention in Pathogenesis

Numerous extrahepatic manifestations have been reported in patients with both acute and chronic hepatitis B (arthralgias or arthritis, skin rashes, glomerulonephritis and neuritis), all of which are present in polyarteritis nodosa (PAN) which is the most unique and spectacular extrahepatic manifestation. In the 1970s, the frequency of PAN due to the hepatitis B (HBV) reached 30%. Immunization programs explain the decrease and it is now down to 7%. PAN usually occurs within 6 months of infection. Clinical manifestations reflect this most classic form of PAN, Hepatic manifestations including, ALT/AST elevations are mild and usually overlooked. Besides HBV, other viruses may be responsible for cases of vasculitides including PAN, HIV, Parvovirus B19, and EBV. Different pathogenic mechanisms have been identified but immune complexes are mainly thought to be responsible. In glomerulonephritis, detailed immunostaining and ultrastructural findings indicate that HBe antigen (Ag) is more likely to be the responsible antigen. In PAN, fewer reports are available and early studies with poorly defined antibodies need to be revisited. Interestingly almost all cases of HBV/PAN are associated with wild-type HBV infection, characterised by HBe antigenemia and high HBV replication, supporting the concept that lesions could result from the deposit of viral Ag/Ab complexes soluble in Ag excess, possibly involving HBe Ag. The recent observation of PAN cases associated with precore mutation which abrogates the formation of HBe Ag challenges this view. It may suggest that other, still undefined, circulating HBV-related Ag(s) distinct from HBe Ag could be involved. Remarkably, none of the HBV/PAN cases or glomerulonephritis exhibit antineutrophil cycoplasmic antibodies (ANCA) reactivity. Viral PAN can now be completely separated from other form of vasculitis mostly autoimmune in nature. Based on the efficacy of antiviral agents in chronic hepatitis B and of plasma exchanges in PAN we combined both therapies to treat HBV PAN. This was associated with swift recovery, even in the most severe forms. The perfect time correlation between inhibition of virus replication and resolution of all bioclinical signs suggest a direct pathogenic role of the virus possibly via immune complexes. Traditional immunosuppressive and steroid therapy should no longer be used for HBV PAN cases.

Remission of Nephrotic Syndrome of HBV-Associated Membranous Glomerulopathy following Treatment with Interferon

We report a 26-year-old homosexual man who developed membranous glomerulopathy with nephrotic syndrome secondary to hepatitis B virus infection and HBe antigenemia. Aminotransferase levels were minimally abnormal, and a liver biopsy showed mild chronic 'persistent' hepatitis. He was initially treated for 4 weeks with human lymphoblastoid alpha-interferon by subcutaneous injection without effect. A second 4-week course of interferon in combination with acyclovir also failed to eradicate HBeAg from the circulation or to reduce the proteinuria. Four years later, he developed new symptomatic hepatitis, with plasma aminotransferases elevated to 200-300 IU/l for more than 4 months. Treatment with interferon was again initiated, and by the 4th month of therapy, he had seroconverted to anti-e status, and cleared the HBeAg from circulation. At the same time, proteinuria significantly dropped from an average of 7 g/day to less than 0.5 g/day. Four years after completion of interferon treatment, he became HBsAg negative and anti-HBsAg reactive while remaining persistently HBeAg negative and anti-HBe positive. He has been free of edema, with normal renal and hepatic function, and his 24-hour protein excretion was less than 0.12 g/day.

Infection with hepatitis A, B, delta, and human immunodeficiency viruses in children receiving cycled cancer chemotherapy.

Serological markers of hepatitis A, B, and Delta and human immunodeficiency viruses were studied in 25 children receiving cancer chemotherapy. Eighty-eight percent had pre-existing HAV immunity which was unaltered by chemotherapy. HDV infection was observed in 8% while HIV was conspicuous by its absence. Active HBV infection, observed in 76% of the children, was asymptomatic in the majority and was accompanied by a high incidence of HBe antigenaemia (57.9%) and its persistence. Pre-existing anti-HBs failed to prevent HBV infection recurrence, which was, however, transient and self-limiting. Multiple blood transfusions and repeated parenteral exposures appeared to be the possible sources of HBV acquisition. Transmission to close contacts was also observed. The study suggests that although HBV vaccine might not be protective against HBV infection in patients receiving cancer chemotherapy, it may prevent its persistence and thereby help in reducing chronic liver disease-related morbidity and a highly infectious reservoir. Strict HBV screening of blood donors, exclusive use of disposable equipment, and vaccination of close contacts of cancer patients is recommended, particularly in HBV endemic third-world countries.

Rapid detection and further characterization of infection with hepatitis B virus variants containing a stop codon in the distal pre-C region

Recently, hepatitis B virus (HBV) replication in the absence of HBe antigenaemia has been attributed to HBV variants with a TAG stop codon in the distal pre-C region associated with one or two point mutations. We describe here a rapid detection method for the diagnosis of such HBeAg-negative HBV variants using selective oligonucleotide hybridization. The entire pre-C region was amplified by the polymerase chain reaction and hybridized under stringent conditions with non-mutated (M0), one (M1) and two (M2) point-mutated oligonucleotide probes. Of the 15 HBeAg-positive (group I) and 20 HBeAg-negative (group II) serum samples studied, 14 samples in group I and one sample in group II hybridized with M0 only and 18 samples in group II hybridized with M1 or M2, or both. The remaining two samples (from groups I and II, respectively) failed to hybridize with any of the three probes. DNA sequencing confirmed mixed distal pre-C sequences in samples hybridizing with more than one probe and also revealed novel mutations in the distal pre-C region of the two samples which failed to hybridize with any of the probes. The latter sample had a +2 frameshift and hence represented a new type of HBeAg-negative HBV variant. This method may therefore prove useful in the diagnosis of infections by HBeAg-negative HBV variants resulting from common mutations in the pre-C region, as well as for the identification of less common variants with novel mutations in the same region.

Elimination of circulatory IgM anti-HSA precedes anti-HBe seroconversion in patients with CAH type B.

The presence of IgM and IgA antibodies with specificity for human serum albumin (HSA) were consecutively analyzed in serum samples from four patients with biopsy verified CAH type B during seroconversion in the HBe/anti-HBe antigen system. Circulatory IgM anti-HSA antibodies were present during HBe antigenemia. The antibody titers fluctuated, decreased, and were finally lost from the circulation. After the disappearance of IgM anti-HSA antibodies, seroconversion to anti-HBe reactivity occurred and a quiescent phase of the disease was established, as judged by normalization of transaminases and absence of circulatory HBV-DNA. IgA anti-HSA antibodies persisted in the circulation after the elimination of IgM anti-HSA and seroconversion to anti-HBe reactivity. For one of the patients, a dramatic increase in titers was followed by elimination of IgA anti-HSA and seroconversion to anti-HBs. The data indicate that the host "self"-component HSA, when associated with "foreign" HBe or HBs antigenic structures, elicit immune responses to HSA, preventing the adequate development of anti-HBe and anti-HBs. The cessation of anti-HSA reactivity, however, seemed to permit subsequent sensitization to HBe and HBs antigenic determinants, as detected by the presence of circulatory antibodies.

Chronic persistent hepatitis. Long-term prospective study on the natural course of the disease.

Ninety-nine patients with chronic persistent hepatitis were followed up in a prospective study for an average of 6.7 years. Fifty-six patients had positive HBV markers; 29 of these patients had persistent HBs antigenemia, and 4 had persistent HBe antigenemia. No patient developed chronic active hepatitis or cirrhosis of the liver. It can be concluded that chronic persistent hepatitis is a benign non-progressive disease that does not require therapy.

A Case of Hepatitis B Virus Associated Nephropathy

Since Combes' first description of hepatitis B virus associated membranous glomerulonephritis, many reports have shown a strong association of hepatitis B virus infection with various types of nephropathies, especially membranous glomerulonephritis and membranoproliferative glomerulonephritis. Recently, the authors experienced a case of membraoproliferative glomerulonephritis, type I in a 16-year-old male patient with persistent HBs and HBe antigenemia. One year prior to admission the patient was told of hepatitis at routine check, but he remained asymptomatic throughout. He was admitted to our hospital with chief complaints of proteinuria and microscopic hematuria found on routine urinalysis. Liver and kidney biopsy revealed chronic persistent hepatitis and membranoproliferative glomerulonephritis type I, respectively. We report a case of hepatitis B virus associated nephropathy with review of the literatures. Considering the endemic nature of hepatitis B virus infection in Korea, the incidence of hepatitis B viurs associated nephropathy would be proportionally high in comparison with those of other countries. So, the importance of this entity merits special consideration in our country. Further study concerning pathogenesis, epidemiology, and treatment may be needed.

TREATMENT OF CHRONIC HEPATITIS DUE TO HEPATITIS B VIRUS

A clearer view of the natural history of chronic hepatitis B virus (HBV) infection has permitted recognition of a phase of viral replication associated with progressive liver damage, and one of absent replication when the disease is inactive and when continued presence of hepatitis B surface antigen (HBs) is due to the integration of viral genes with the host genome. These two phases can be identified by HBe antigenaemia and anti-HBe, respectively. Several active antiviral drugs are available and may significantly benefit certain HBe Ag-positive groups. The antiviral activity of vidarabine and its analogues and of α-interferons is established, and insight is being gained into factors that predict response. In general, results depend on duration of infection, and integrity of the patient's immune response. Anti-HBe positive carriers usually need no treatment, but in those with continuing low-level HBV replication or delta superinfection antiviral therapy, although of unproven value, may be tried. In patients without HBV or hepatitis delta virus (HDV) replication who have signs of active disease, immunosuppressants may be tried with benefit.

Detection of two forms of HBeAg (free‐and IgG‐bound HBeAg) in patients with HBe antigenemia using staphylococcus bearing protein A

Protein A-bearing Staphylococcus aureus organisms (STA) were used to separate free HBeAg from IgG-bound HBeAg. Free HBeAg was detected in the supernate while IgG-bound HBeAg could be liberated from the pellets using MgCl2 or a glycine buffer. HBeAg was determined by radioimmunoassay and the results expressed as patient's cpm/normal control's cpm ratio (S/N ratio). This ratio was demonstrated to be proportionate to the antigen concentration and used as a titer of HBeAg. Sera of 40 HBsAg-negative healthy volunteers and 82 HBeAg-positive patients who were either asymptomatic HBsAg carriers or had various diseases including chronic persistent hepatitis (CPH), chronic active hepatitis (CAH), and renal disease undergoing hemodialysis, were tested for free HBeAg and IgG-bound HBeAg. Patients with CAH from two different countries were compared. Free HBeAg was detected in all patients but one, IgG-bound HBeAg was detected with similar prevalences (from 56% to 67%) in HBsAg asymptomatic carriers, hemodialysis patients, CPH and Italian CAH patients. In contrast, all CAH patients from New York, who had frequently been exposed to HBV infection, had detectable levels of IgG-bound HBeAg, with the highest S/N ratios observed in the study, and frequently showed an unfavorable outcome. In AVH due to HBV and delta agent co-infection, IgG-bound HBeAg was detected in two of four patients only in the initial stage of the disease. The data reported indicate that a separate determination of free- and IgG-bound HBeAg may have clinical value.

Detection of serum HBV-DNA by molecular hybridisation: Correlation with HBeAg/Anti-HBe status, racial origin, liver histology and hepatocellular carcinoma

The relationship of the presence of hepatitis B virus (HBV) DNA in serum, a measure of HBV-replication, to HBeAg/anti-HBe status has been examined. In Northern Europe, there is a strong positive correlation between the presence of HBV-DNA and HBe antigenaemia and a negative correlation with the presence of anti-HBe. These associations are less marked in patients from Southern Europe, Africa, the Middle and Far East. When HBV-DNA is present in the serum of anti-HBe carriers, it is usually associated with the presence of severe liver disease or carcinoma. Forty percent of patients with hepatocellular carcinoma had evidence of continuing HBV replication.

Analysis of the molecular state of HBV-DNA in the liver and serum of patients with chronic hepatitis or primary liver cell carcinoma and the effect of therapy with adenine arabinoside.

The pattern of replicative intermediates seen in the liver of HBe antigen and antibody positive patients was determined. During the phase of HBe antigenaemia the 3.2 Kb species of HBV-DNA (complete HBV genome) is present in the liver but not in the serum. When HBe antigen to antibody seroconversion occurs, either spontaneously or during antiviral therapy, the 3.2 Kb and lower molecular weight intermediates disappear from the liver and the 3.2 Kb band appears transiently in the serum. Integrated HBV-DNA was found in one of 15 patients during the period of HBe antigenaemia and in three of seven patients in the HBe antibody positive phase of the chronic infection before detection of primary liver cell carcinoma. Integrated sequences were found in tumour tissue of two patients with primary liver cell carcinoma who were anti-HBc positive but were absent from the tissues of two patients developing primary liver cell carcinoma at a late stage of autoimmune liver disease. These studies suggest that integration of the HBV genome occurs rarely or in only a small proportion of hepatocytes during the early (HBe antigen positive) phase of infection in Caucasians. They also show that not all primary liver cell carcinomas necessarily contain HBV-DNA.

More confined indications for use of combined passive and active immunization for preventing perinatal development of hepatitis B virus carrier-state - based on the natural history of hepatitis B virus-vertical transmission

To find more confined criteria for use of passive and/or active immunization for preventing perinatal development of hepatitis B virus (HBV) carrier-state than maternal HBe antigenemia, maternal HBsAg-titers (R-PHA) around delivery and infantile HBeAg-titers-(EIA) are discussed. No children whose maternal HBsAg-titers around delivery were lower than 3 6 developed carrier-state in spite of maternal HBe antigenemia. In addition, at age 2 months serum HBeAg-titers of 6 children who had acquired persistent HBsAb were lower than 25, while those of 5 children who had developed carrier-state were higher than 70. These findings may contribute to the establishment of more confined indications for the administration of HBIG and/or HB vaccine to the children born to HBeAg-positive carrier women, saving not only HBIG and HB vaccine but all accompanied efforts of both patients and medical staff as well.

Virus receptors for polymerized human albumin: a prognostic marker in HBeAg-positive chronic hepatitis type B?

Seventeen out of 30 patients with chronic hepatitis type B with hepatitis B e antigen (HBeAg) in serum remained persistently positive for e antigen, while 13 seroconverted to antibody (anti-HBe) when followed over a period of one to five years. Initial levels of serum hepatitis B virus (HBV) markers, such as the hepatitis B surface antigen (HBsAg), HBeAg, and HBV-DNA polymerase (HBV-DNAP) were similar in the two groups of patients, while initial titres of the HBsAg-associated receptor for polymerized human serum albumin (pHSA), recently identified on HBV particles, were significantly higher in the patients who remained HBeAg positive (mean titre +/- SD = 2(-7.00) +/- 2(-3.2)) compared to the cases who eventually seroconverted to anti-HBe during the follow-up (2(-2.54) +/- 2(-2.14) P less than 0.001). A receptor titre above 1:64 by haemagglutination was highly predictive of persistence of HBeAg, suggesting that in patients with HBeAg-positive chronic hepatitis testing for the HBsAg-associated pHSA receptor may be useful in predicting the duration of HBe antigenaemia, with relevant clinical and prognostic implications.

THE ROLE OF HBe ANTIGEN IN THE PATHOGENESIS OF HB ASSOCIATED MEMBRANOUS GLOMERULONEPHRITIS

The purpose of the present study is to investigate the role of HBe antigen in the pathogenesis of membranous glomerulonephritis (MGN) in Japanese children. Of 364 children who underwent renal biopsy from 1976 through 1978 in our institution, 11(10 male, 1 female, ages 1 - 14 years) were found to be HBs antigen carriers and comprise the subject of the present study. All of them exhibited some abnormality on a routine urinalysis at one time during the course. Serum HB associated antigens were studied in all. A routine light, fluorescent and electron microscopic study was performed on 14 biopsied specimens from the 11children. In addition, direct immunofluorescence for HBsAg, HBeAg & HBcAg was done in the majority of the specimens. The results are summerized in the table. The 14 specimens were divided into 2 groups depending upon clinical manifestations at the time of renal biopsy (Group A: 9 specimens from 9 children with nephrotic syndrome or heavy proteinuria, Group B: 5 specimens from 4 children with minimal hematuria). All of the Group A specimens showed MGN stage II or III and the Group B showed either minimal changes or MGN stage IV. In all of the 5 Group A specimens studied HBeAg immune complex was present along the glomerular capillary wall, but not present in any of the 4 Group B specimens. However, HBsAg was demonstrable in only one of the 11 specimens tested. At the time of renal biopsy, circulating HBeAg was studied in 8 children (Group A: 4, Group B: 4). It is of interest that 2 of the 4 Group B children were found to have HBe antigenemia. The above data suggests there is a highly significant relationship between glomerular HBeAg immune complex and the development of MGN. We believe that most cases of HB associated nephropathy seen in the Japanese children are HBeAg mediated MGN.