HBeAg-positive Carriers Research Articles

Investigating the Correlation between Semi quantitative HBeAg and HBV DNA Levels: Validating HBeAg Quantification as a Marker in Asymptomatic HBeAg-Positive Carriers

Investigating the Correlation between Semi quantitative HBeAg and HBV DNA Levels: Validating HBeAg Quantification as a Marker in Asymptomatic HBeAg-Positive Carriers

Common Genetic Variants of Response to Hepatitis B Vaccines Correlate with Risks of Chronic Infection of Hepatitis B Virus: A Community-Based Case-Control Study.

Hepatitis B (HB) vaccination effectively reduces the risks of chronic infection with the hepatitis B virus (HBV). It is unknown whether there is a common genetic determinant for response to the HB vaccine and susceptibility to chronic HBV infection. This case-control study, which included 193 chronic HBV carriers and 495 non-carriers, aimed to explore the effects of the most significant single nucleotide polymorphisms (SNPs) in response to the HB vaccine on the risks of chronic HBV infection. Out of 13 tested SNPs, the genotype distributions of four SNPs at the human leukocyte antigen (HLA) class II region, including rs34039593, rs614348, rs7770370, and rs9277535, were significantly different between HBV carriers and non-carriers. The age-sex-adjusted odds ratios (OR) of chronic HBV infection for rs34039593 TG, rs614348 TC, rs7770370 AA, and rs9277535 AA genotypes were 0.51 (95% confidence interval [CI], 0.33-0.79; p = 0.0028), 0.49 (95% CI, 0.32-0.75; p = 6.5 × 10-4), 0.33 (95% CI, 0.18-0.63; p = 7.4 × 10-4), and 0.31 (95% CI, 0.14-0.70; p = 0.0043), respectively. Multivariable analyses showed that rs614348 TC and rs7770370 AA genotypes were significantly independent protectors against chronic HBV infection. The multivariable-adjusted ORs for subjects with none, either one, or both of the protective genotypes were 1.00 (referent), 0.47 (95% CI: 0.32-0.71; p = 3.0 × 10-4), and 0.16 (95% CI: 0.05-0.54; p = 0.0032), respectively. Among eight HBeAg-positive carriers, only one of them carried a protective genotype. This study shows that response to the HB vaccine and susceptibility to chronic HBV infection share common genetic determinants and indicates that HLA class II members are the main responsible host genetic factors.

Epidemiology and clinical management of HIV-HBV coinfected patients in a large AIDS Reference Center in Belgium

ABSTRACTObjectives: Human immunodeficiency virus (HIV) and hepatitis B virus (HBV) are both worldwide health concerns with similar routes of transmission and no curative treatment to date. Coinfection is associated with increased morbidity and mortality. We aim to provide epidemiological data about HIV-HBV coinfected patients and asses if management of patients following European recommendation (EACS) was achieved in a large AIDS Reference Center in Belgium.Methods: Retrospective review of the HIV database of Saint-Pierre University Hospital in Brussels (Belgium) focusing on HIV-HBV coinfected patients in active follow-up. We classified patients in six serological profiles: (A) patients with active chronic HBV infection (HBsAg positive and HBeAg positive), (B) patients with persistent chronic HBV infection (HBsAg positive and HBeAg negative), (C) patients with isolated core antibody (isolated anti-HBc positive), (D) patients with resolved HBV infection (anti-HBc positive and anti-HBs positive), (E) vaccinated patients (anti-HBs positive), and (F) patients with all above markers negative. Chronic HBV infection (cHBV) was defined by two positive hepatitis B surface antigens (AgHBs positive) with at least 6-month intervals and chronic HBeAg positive group by a positive AgHBe (Ag HBe positive). We reviewed individual files of HIV-HBV chronically coinfected patients to assess if European recommendations in terms of HBV coinfection management were adequately followed in our center.Results: Among 2601 HIV-infected patients in active follow-up, 98 (3.8%) were chronically infected with HBV. Median age of chronically coinfected patients was 46 years with male predominance and heterosexual Africans representing the majority. Among the chronically coinfected patients, 33.7% were HBeAg positive carriers. Mean HBV DNA and ALT/AST were significantly higher in the chronic HBeAg positive (cHBeAg positive) patients compared to chronic HBeAg negative patients (cHBeAg negative). Nearly 95% of the cHBV patients were treated with two anti-HBV drugs (99% for the cHBeAg positive group), with 79% having Tenofovir (TDF) in their antiretroviral treatment history. 8% were screened for hepatitis D virus (HDV) antibodies. Liver fibrosis, upper endoscopy and alpha-foetoprotein were assessed at least once in the last 5 years in 32%, 31% and 32% of cHBV patients respectively. cHBeAg positive patients were not significantly monitored closer except for liver fibrosis assessment in 52% (p < 0.0017).Conclusion: The prevalence of cHBV coinfection in the Saint-Pierre HIV cohort is lower than in neighboring European countries. Hepatic monitoring should be reinforced in our cHBV and cHBeAg positive patients because of higher risk of progression to cirrhosis progression and hepatocellular carcinoma.

Detection of Hepatitis B Genotypes in HBsAg &amp; HBeAg Carriers

Objective:- Hepatitis B is noteworthy medical issues that may include the late continuation of liver cirrhosis and hepatocellular carcinoma. The present study aimed for the detection and diffrentiation of Hepatitis B virus HBsAg inactive non-replicative carriers, HBeAg-positive inactive replicative carriers, active carriers &amp; HBeAg-negative chronic hepatitis B by Real Time PCR and their genotyping&#x0D; Methods: This research conducted on 245 positive for HBsAg, 118 (48.16 %) were male and 127 (51.84%) were female patients, which was performed in central research station labortory of Microbiology at netaji subhash Chandra Bose subharti Medical College and Hospital, Meerut Between march 2016 to November 2017 The sera were separated and screened for HBsAg by ELISA kit. Positive samples for HBsAg were tested for HBeAg ELISA kit and DNA Viral load then sequenced for genotying&#x0D; Results:. Of the 245 HBsAg Positive case 55 (1.12%) were HBeAg positive. In 16 PCR positive and HBV genotyping, In HBsAg inactive Non-Replicative 37.5% (n=6) genotype-B and 6.25% (n=1) genotype-A, In HBeAg inactive Replicative 12.5% (n=2) genotype-B and 12.5% (n=2) genotype-A and In HBeAg Active Chronic Hepatitis B 18.75% (n=3) genotype-B and 12.5% (n=2) genotype-A were detected&#x0D; Conclusions: Management strategy, using HBsAg, HBeAg and HBV DNA viral load, seems adequate for the confirmation and diffrentiation of Hepatitis B virus inactive, active carriers &amp; HBeAg-negative chronic hepatitis B patients and genotype B was more prevalent in comparission to genotype A. Distribution of carriers &amp; genotypes, help physicians to prescribe proper antiviral/interferon therapy according to current genotyping pattern in this region&#x0D; Keywords: Hepatitis B virus, Carrier State, HBsAg, HBeAg, RT-PCR

Heparin at physiological concentration can enhance PEG-free in vitro infection with human hepatitis B virus

Hepatitis B virus (HBV) is a blood-borne pathogen responsible for chronic hepatitis, cirrhosis, and liver cancer. The mechanism of HBV entry into hepatocytes remains to be investigated. Recently, sodium taurocholate cotransporting polypeptide (NTCP) was discovered as a major HBV receptor based on an in vitro infection system using NTCP-reconstituted HepG2 cells. However, this infection system relies on the compound polyethylene glycol (4% PEG), which is not physiologically relevant to human infection. High concentration of heparin has been commonly used as an inhibitor control for in vitro infection in the field. Surprisingly, we found that heparin at physiological concentration can enhance HBV infection in a PreS1-peptide sensitive, NTCP-dependent manner in both HepaRG and HepG2-NTCP-AS cells. O-sulfation of heparin is more important for the infection enhancement than N-sulfation. This system based on the HepG2-NTCP-AS cells can support in vitro infection with HBV genotypes B and C, as well as using serum samples from HBeAg positive and negative chronic carriers. In summary, our study provides a PEG-free infection system closely resembling human natural infection. In addition, it points to a future research direction for heparin and heparin-binding host factor(s) in the blood, which are potentially involved in viral entry. To our knowledge, this is the first soluble and circulatory host factor which can enhance HBV in vitro infection.

HBeAg Status among Chronic Hepatitis B Patients at A Referral Hospital of Bangladesh

Background: HBeAg status in chronic hepatitis B patients is important for outcome and treatmentObjective: The purpose of the present study was to see the status of HBeAg Chronic Hepatitis B (CHB) patients.Methodology: This cross sectional study was conducted in the Department of Virology at Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka between July 2010 to June 2011. Serologically diagnosed CHB patients were enrolled for the study. The HBV DNA was quantified. Samples were tested for HBeAg with ELISA kit.Results: A total of 200 serologically diagnosed CHB patients were enrolled for the study. Among the total study population, HBeAg positive CHB patients were 74(37%) cases and HBeAg negative patients were 126 (63%) cases. Among the HBeAg negative patients, viral load was less and patients were significantly older. The mean viral load of HBeAg positive and HBeAg negative was 64012042 and 2.83i2.55 respectively. HBV DNA was a more reliable indicator of the presence of virus than HBeAg, and was detected in 98.65% (73/74) HBeAg positive carriers, and in 66.67% (84/126) HBeAg negative patients.Conclusion: HBeAg negativity is more prevalent among the CHB patients in Bangladesh.J Shaheed Suhrawardy Med Coll, 2014; 6(2):60-63

A cohort study of hepatitis B virus carriers progressing to chronic hepatitis in Jiangsu province, 2012-2014

Objective: To understand epidemiological characteristics and influencing factors of hepatitis B virus (HBV) carriers progressing to chronic hepatitis B in Jiangsu, and provide evidence for regular management of HBV carriers. Methods: From 2012 to 2014, a cohort study was conducted among the HBV carriers in an area in Jiangsu province. Association between HBV carriers' demographics, HBeAg level and HBV DNA detection result and incidence of chronic hepatitis B was analyzed by Cox regression analyses. Results: In 2012, a total of 4 069 HBV carriers aged (52.0±12.8) years were surveyed. By 2014, chronic hepatitis B had developed in 1 444 cases, with the rate of 21.0/100 person-years. Cox regression analysis indicted that, in addition to gender, HBV DNA level was an important influencing factor for the incidence of chronic hepatitis B (P<0.05). In HBeAg positive carriers progressing to chronic hepatitis, 40.7% had HBeAg seroconversion, and reversion of HBeAg occurred in 1.7% of HBeAg negative carriers. Conclusion: Chronic hepatitis B developed in more than 1/5 (21.0/100 person-years) of HBV carriers in the surveyed area in Jiangsu. It is suggested to conduct regular ALT, HBV DNA detection and B ultrasonic examination in HBV carriers for the early detection of chronic hepatitis B.

Cellular immune responses in patients with hepatitis B surface antigen seroclearance induced by antiviral therapy

BackgroundThe mechanisms by which chronic hepatitis B is completely resolved through antiviral therapy are unknown, and the contribution of acquired T cell immunity to hepatitis B surface antigen (HBsAg) seroclearance has not been investigated. Therefore, we measured the T-cell responses to core and envelope antigens in patients with HBsAg seroclearance.MethodsFourteen subjects with HBsAg seroclearance following antiviral treatment for chronic hepatitis B, 7 HBeAg-positive immunotolerant HBV carriers and 9 HBeAg-negative inactive HBsAg carriers were recruited. HBV-specific T-cell responses to recombinant HBV core (rHBcAg) and envelope (rHBsAg) proteins and pools of core and envelope peptides were measured using an ELISPOT assay detecting interferon-gamma and intracellular cytokine staining (ICS) assays detecting interferon-gamma or interleukin 2.ResultsInterferon-gamma ELISPOT assays showed a low frequency of weak responses to the rHBsAg and S peptide pool in the HBsAg seroclearance group, and the response frequency to the rHBcAg and the C peptide pool was higher than to the rHBsAg (P < 0.001) and S peptide pool (P = 0.001) respectively. A higher response frequency to C than S peptide pools was confirmed in the interferon-gamma ICS assays for both CD4+ (P = 0.033) and CD8+ (P = 0.040) T cells in the HBsAg seroclearance group. The responses to C and S antigens in the inactive carriers were similar.ConclusionsThere was a low frequency of CD4+ and CD8+ T cell immune responses to envelope antigens in Chinese subjects with HBsAg seroclearance following antiviral therapy. It is unlikely that these immune responses are responsible for HBsAg seroclearance in these subjects.

Immune response of peripheral blood mononuclear cells to antigenic determinants within hepatitis B core antigen in HB virus-infected man

Residues 72-146 within hepatitis B core Ag (HBcAg) represent T-cell recognition site in HB-virus-infected man. This study was undertaken to define critical residues involved in the immunogenicity of dominant T-cell determinants of HBcAg. For this purpose, p120-131 and its analog (p120-131 [A] containing alanine substitutions at residues 122 and 125, which were identified as epitopic residues in mice, were synthesized. These peptides and recombinant HBcAg were analyzed for their ability to stimulate peripheral blood mononuclear cells (PBMC) from 25 patients with chronic HBV infection and three patients with acute hepatitis B. PBMC from 18 out of 28 patients showed significantly increased IFN-gamma production and proliferative response in the presence of recombinant HBcAg. Eight patients responded to the two peptides, while 12 patients did not. Four patients responded only to p120-131, and four displayed a response only to p120-131 [A]. The responses to the two peptides were similar among HBeAg-positive and anti-HBe-positive patients, and did not depend on disease activity, except for HBeAg-positive asymptomatic carriers in whom there was no response to any additive. These results indicate that immune responses to p120-131 and its analog were similar in our patient groups. The dominant epitopic residues in this region of HBcAg may differ between man and mouse.

A new clade of hepatitis B virus subgenotype F1 from Peru with unusual properties

There are eight genotypes A-H of hepatitis B virus (HBV). Most genotypes are further divided into subgenotypes. Genotypes and subgenotypes influence the natural course of infection and therapy. We analysed nine sera from HBV carriers from Peru. Using the small hepatitis B surface protein HBs, all samples could be grouped to genotype F. Sequencing of three complete Peruvian genomes showed that HBV from Peru belongs to subgenotype F1. Two of the genomes from HBeAg positive carriers coded surprisingly for a stop codon in the polymerase-ORF leading to a translational stop after 213 and 214 aa, respectively. The third isolate from an HBe Ag positive carrier had three deletions: aa 1-53 and aa 111-142 in preS. In addition nt. 2002-2087 in the HBc-ORF were deleted, leading to an HBc starting at aa 66.

Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia

Background/AimThe study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials.MethodQuantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers.ResultsThe qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range (1.5 × 103 to 1.5 × 108 copies/mL) and correlated well with those from a commercial assay (r = 0.91, (p < 0.001). Viral load levels dropped dramatically but temporarily during and after a short course of lamivudine therapy. HBV DNA was a more reliable indicator of the presence of virus than HBe antigen and was detected in 77.0% (161/209) of HBeAg negative and in all HBeAg positive carriers.ConclusionThis method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.

Epidemiology of precore mutants of hepatitis B in the United Kingdom.

A point mutation assay was used to study the codon 28 and codon 1 precore mutant status of 310 chronic hepatitis B carriers (82 HBeAg positive and 228 HBeAg negative). Fourteen of 228 (6%) of HBeAg negative carriers had high levels of serum HBV DNA. Nine of these were explained by precore variants, three by core promoter variants, and two were not explained by recognised precore changes. Nested PCR detected serum HBV DNA in 36% (82/228) of HBeAg negative carriers and 63% (52/82) of these had precore variants. Four of 82 (4%) of the HBeAg positive carriers had precore variants, all as mixed mutant/wild type populations and evidence indicated that these carriers were seroconverting. Overall 23% (52/228) of HBeAg negative carriers had both serum HBV DNA and codon 1 or 28 precore mutations. A sexual transmission event from an HBeAg negative carrier with a relatively low serum HBV DNA level (10(4)-10(6) genome copies/ml) and only core promoter mutations was observed. Despite high rates of variant carriage in the antenatal sub-group perinatal transmission was not observed. The results of direct sequencing on 45 carriers validated the point mutation assay and also showed that codon 28 mutations were only seen in carriers with the genotype CCT at codon 15. For the Caucasian population a higher prevalence of codon 28 mutations (13/25 or 52%) than expected was seen. Liver biopsy data indicated that there was no link between the presence or absence of precore mutants and the severity of liver disease.

Association between frequency of amino acid changes in core region of hepatitis B virus (HBV) and the presence of precore mutation in Japanese HBV carriers.

To elucidate the relationship between the frequency of core mutations and precore mutation of hepatitis B virus (HBV) in Japanese HBV carriers, we investigated the nucleotide sequence of the precore/core region of HBV in 26 Japanese HBV carriers [15 who were HBe antigen-negative (HBeAg-) and 11 who were HBeAg-positive (HBeAg+)]. The number of amino acid changes (5.9 +/- 3.8) in the core region of HBV in HBeAg-carriers was significantly greater than that in the HBeAg+ carriers (1.5 +/- 1.0; P < 0.005). The precore stop codon mutation was found in 93.3% of HBeAg-negative HBV carriers, while no precore mutation was found in the HBeAg-positive HBV carriers, suggesting that the frequency of core mutations may be associated with the presence of the precore stop codon mutation. However, there was no significant difference in the frequency of amino acid changes among HBeAg-HBV carriers. The mean number of core amino acid changes of liver cirrhosis patients, chronic active hepatitis patients, chronic persistent hepatitis patients, and asymptomatic carriers were 2.7 +/- 1.5, 6.0 +/- 2.2, 4.7 +/- 1.2, and 8.4 +/- 5.3, respectively. We detected hot spots for core mutations, which showed characteristic localizations and specific substitutions: Gly-87, Leu-97, and Thr-130 were detected exclusively in patients with chronic liver disease with or without HBeAg. To address further the relationship between frequency of core mutations and the presence of the precore stop codon mutation, we investigated the precore/core nucleotide sequence serially along with seroconversion in three patients with chronic hepatitis B in whom the hepatitis either became inactive or remained active after the seroconversion. Emergence of the precore stop codon mutation and a significant increase in core amino-acid changes after seroconversion were noted in all three patients. Our results suggest a close association between the frequency of core amino acid changes and the presence of the precore stop codon mutation; some characteristic core mutations may be associated with the clinical course of chronic hepatitis B in Japanese patients.

Detection of Serum Hepatitis B Virus DNA According to HBV Markers in Chronic Hepatitis B Liver Disease

The identification of serum HBV DNA is very important for the assessment of the disease activity in persistent infection, for the evaluation of the infectivity of an individuals blood. The dot blot, however, has limited sensitivity and sometimes inconsistent with other serological markers and clinical settings. Using the most important recent advance in molecular biology, the polymerase chain reaction(PCR), specific DNA sequences can be amplified more than a million-fold in a few hours and with this technique the detection of the extreme low level of DNA is possible. This study was to determine sensitivity of the PCR for the detection of serum HBV DNA in comparison with dot blot analysis and to investigate the serum HBV DNA status and clinical significance of PCR in patients with chronic HBsAg positive liver disease. The subjects of this study were 17 patients with asymptomatic HBsAg carriers(9 HBeAg positive patients, 8 anti-HBe positive patients), 91 chronic hepatitis B(50 HBeAg positive patients, 41 anti-HBe positive patients), 57 liver cirrhosis(21 HBeAg positive patients, 36 anti-HBe positive patients), 27 hepatocellular carcinoma(10 HBeAg positive patients, 17 anti-HBe positive patients). The results were summerized as following; The detection rates of HBV DNA by dot blot, PCR were 58.9%, 72.2% in HBeAg positive patients, 34.3%, 53.9% in anti-HBe positive patients. The detection rates of HBV DNA by PCR in HBeAg negative patients were 25.0% in asymptomatic HBsAg carriers, 61.0% in chronic hepatitis B, 52.8% in liver cirrhosis, 52.9% in hepatocellular carcinoma. The positive rate for HBV DNA is a significant difference between HBeAg positive and negative asymptomatic HBsAg carriers, but not significantly difference in other groups. In conclusions, this study confirmed that the PCR is much more sensitive than the dot blot analysis in detecting the HBV DNA in the sera of patients with chronic liver disease. The presence of HBV DNA in the serum was detected by PCR with higher sensitivity and it suggested that active viral replication is still going on in most patients with chronic HBsAg positive liver disease irrespective of HBeAg/anti-HBe status, and PCR may be used as a prognostic factor in asymptomatic HBsAg carriers.

Prevalence of precore‐defective mutant of hepatitis B virus in HBV carriers

Two hundred and seventy-three serum specimens from hepatitis B virus (HBV) carriers were examined for the presence of a characteristic one point mutation at nucleotide (nt) 1896 from the EcoRI site of the HBV genome in the precore region (the preC mutant) using restriction fragment length polymorphism (RFLP) analysis. This assay approach could detect preC mutants or wild-type sequences when either form constituted more than 10% of the total sample. Overall, 65.5% (76/116) of HBeAg-positive carriers had only the preC wild-type. All HBeAg-positive asymptomatic carriers (n = 14) had only the preC wild-type. In patients with chronic hepatitis B and in anti-HBe-positive asymptomatic carriers, increased prevalence of the preC mutant was associated with the development of anti-HBe antibodies and normalization of the serum alanine aminotransferase concentration. Furthermore, 27 (29.0%) of 93 HBeAg-negative carriers had unexpectedly preC wild-type sequences only. Direct sequencing of the HBV precore region of HBV specimens from 24 patients revealed no mutation at nt 1896, supporting the specificity of the RFLP analysis. These results suggest that RFLP analysis was accurate for the detection of the preC mutation and that the absence of serum HBeAg cannot be explained solely by the dominance of the preC mutant.

Recent results in the treatment of chronic B virus hepatitis

Of the drugs evaluated to date for the therapy of chronic hepatitis B, only alpha-interferons have gained wide acceptance as single-agent therapy. In HBeAg-positive carriers, treatment must be carried out for 4-6 months on an alternate-day basis and dosage should be not less than 5 million U/m2 of body surface. Oriental patients, children, immunodeficient and highly viremic patients are less likely to respond. Patients given combination therapy (with steroids, antivirals, stimulators of the immune system) do not appear to benefit from the association in comparison with treatment with interferon alone. In most patients (60-80%) with atypical chronic type B infections (anti-HBe-positive, HBV-DNA-positive, intrahepatic HBcAg), HBV-DNA becomes negative and transaminases normalize during interferon treatment; however, many (80%) experience a relapse of viremia and disease during follow-up. Side effects are usually minor (flu-like symptoms), but in a minority of patients, major adverse events have also been reported. alpha-Interferon is effective in inhibiting viral replication in a significant number of patients with chronic type B hepatitis, but new therapeutic regimens and a better selection of patients are needed in order to induce persistent remissions and reduce the cost/benefit ratio.

Ｂ型肝炎ウイルス（ＨＢＶ）キャリアクリニック 血液センターにおける１０年間の経験から

We established a special clinic (in 1985) for asymptomatic HBV carriers detected through blood donation. The main purpose of this clinic was the early detection and treatment of chronic hepatitis. In 10 years 3, 367 notified carriers detected from over two million blood donors visited our clinic after attending an orientation seminar.The HBsAg-positive rate among first-time donors decreased from 3.0% to 1.2% during these 10 years. The overall HBeAg-positive rate in clinic visitors was 13.2%, and decreased with age. Elevated GPT levels were seen in 6.0% of clinic visitors, with a higher rate in HBeAg-positive carriers. HBsAg as tested with EIA disappeared in 40 cases (1.2%). HBeAg disappeared in 23 cases with elevated GPT. HCV antibody was detected in 16 cases.Carriers with elevated GPT or space-occupying lesions in the liver as detected by echography were referred to specialized hospitals. Of these, 166 (4.9%) cases were diagnosed as HBV-related liver disease, including 5 cases of hepatoma. Thus, regular follow-up in our HBV carrier clinic strongly contributed to the medical care of asymptomatic HBV carriers.

Mutations in the core promoter/enhancer II regions of naturally occurring hepatitis B virus variants and analysis of the effects on transcription activities.

The regulatory regions for transcription and replication of several hepatitis B virus (HBV) genomes from 19 patients having various forms of HBV infection were sequenced. Predominant mutations were found to occur naturally in nucleotide positions 1762 (A to T) and 1764 (G to A) in chronic hepatitis patients and in asymptomatic carriers after seroconversion, but were not observed in HBeAg-positive healthy carriers. Since these positions were located in the basic core promoter and the overlapping enhancer II regions situated within the core upstream region, transcriptional activity was examined by chloramphenicol acetyltransferase assay to determine if there was a possible difference between the mutant and wild-type HBV. However, no significant difference was detected upon comparison of the promoter and enhancer activities between mutant and wild-type HBV.

HBsAg subtypes among HBsAg carriers in Okinawa, Japan. Evidence of an important relationship in seroconversion from HBeAg to anti-HBe

In Okinawa, Japan 1261 hepatitis B surface antigen (HBsAg) carriers were investigated for clinical differences among HBsAg subtypes. Among the 854 for whom they could be determined, subtype adw was found in 604 (70.7%), adr in 232 (27.2%) and others in 18 (2.1%). The overall prevalence of hepatitis B e antigen (HBeAg) was significantly lower in subtype adw (11.9%) than in adr (17.7%) (P < 0.01). The mean age of HBeAg-positive carriers was significantly lower in adw (20.4 years) than in adr (26.9 years) (P < 0.05). Seroconversion from HBeAg to antibody to HBeAg (anti-HBe) occurred in 43 carriers. The seroconversion rate was higher in adw (43.1%) than in adr (29.3%) and the seroconversion age was younger in adw (22.7 years) than in adr (27.9 years). These results suggest that carriers with subtype adw tend to seroconvert from HBeAg to anti-HBe at a higher rate and a younger age than do those with adr. HBsAg subtypes may be closely associated with the HBeAg/anti-HBe status.

Immune recognition of linear antigenic regions within the hepatitis B pre-C and C-gene translation products using synthetic peptides.

The antibody recognition of linear regions within the amino acid (aa) sequence of hepatitis B (HB) core antigen (HBcAg), e antigen (HBeAg), and pre-C region was investigated in 46 patients infected with hepatitis B virus (HBV), and one immunized rabbit. Peptide analogues were synthesized to cover the complete product of the C-gene, including the pre-C region using various synthetic methods. Two carriers of hepatitis B surface antigen (HBsAg) with anti-HBe, recognized pin-bound decapeptides covering amino acid (aa) 76-83 of HBc/eAg, and the most essential residues were found to be Asp78, Pro79, Arg82, and Asp83. Pre-C peptides were recognized by IgG1 or IgG3 in sera from two out of ten cases with acute HB, in four out of twelve sera from HBeAg-positive carriers of HBsAg, and in two out of twelve sera from anti-HBe-positive carriers of HBsAg. Two sera from the cases of acute HB showed strong reactivity of the IgG3 isotype with HBc/eAg peptides 61-85. Five of the sera from HBeAg-positive carriers of HBsAg were weakly reactive with peptides 41-60, 61-85, 121-140, and/or 141-160. Eight of the sera from anti-HBe-positive carriers of HBsAg recognized aa 121-140 of HBc/e with IgG1, IgG3, and/or IgG4 isotypes. IgG from one immunized rabbit recognized peptides 1-20, 61-85, and 71-90, and the T-cells recognized peptides 1-20 and 71-90. Thus, human and rabbit antibodies recognize linear antigenic regions within the pre-C, and within regions 1-20, 41-60, 61-85, 121-140, and 141-160 of HBcAg.(ABSTRACT TRUNCATED AT 250 WORDS)