Basal core promoter and precore mutations among hepatitis B virus circulating in Brazil and its association with severe forms of hepatic diseases

BACKGROUND In Brazil, few studies have investigated the prevalence of infection with the precore (PC) and basal core promoter (BCP) mutants of the hepatitis B virus (HBV). OBJECTIVES This study aimed to analyse the frequency of PC and BCP mutations among patients infected with HBV and to evaluate the association between the variants and advanced hepatic disease. METHODS A total of 161 patients infected with HBV were studied. To identify PC and BCP mutations, a 501-bp fragment of HBV DNA was amplified and sequenced. FINDINGS PC and BCP regions from HBV strains were successfully amplified and sequenced in 129 and 118 cases, respectively. PC and BCP mutations were detected in 61.0% and 80.6% of the cases, respectively. The A1762T/G1764A variant was identified in 36.7% of the patients with grade 1 and 2 liver fibrosis (29/79) and in 81.8% of the patients with grade 3 and 4 liver fibrosis (9/11) (p < 0.01); in 76.9% of the patients with cirrhosis (10/13) and in 38.1% of the patients without cirrhosis (40/105) (p = 0.01); and in 77.8% of the patients with hepatocellular carcinoma (HCC) (7/9) and in 39.4% of the patients without HCC (43/109) (p = 0.03). MAIN CONCLUSIONS A high prevalence of HBV PC and BCP mutants was found. The A1762T/G1764A variant was independently associated with advanced forms of liver fibrosis, hepatic cirrhosis, and HCC.

Hepatitis B virus genotypes and mutations in the basal core promoter and pre-core/core in chronically infected patients in southern Brazil: a cross-sectional study of HBV genotypes and mutations in chronic carriers

Introduction In Brazil, little data exist regarding the distribution of genotypes in relation to basal core promoter (BCP) and precore/core mutations among chronic hepatitis B virus (HBV) carriers from different regions of the country. The aim of this study was to identify HBV genotypes and the frequency of mutations at the BCP and precore/core region among the prevalent genotypes in chronic carriers from southern Brazil. Methods Nested-polymerase chain reaction (nested-PCR) products amplified from the S-polymerase gene, BCP and precore/core region from 54 samples were sequenced and analyzed. Results Phylogenetic analysis of the S-polymerase gene sequences showed that 66.7% (36/54) of the patients were infected with genotype D (D1, D2, D3), 25.9% (14/54) with genotype A (A1, A2), 5.6% (3/54) with subgenotype C2, and 2% (1/54) with genotype E. A comparison of virological characteristics showed significant differences between genotypes A, C and D. The comparison between HBeAg status and the G1896A stop codon mutation in patients with genotype D revealed a relationship between HBV G1896A precore mutants and genotype D and hepatitis B e antigen (HBeAg) seroconversion. Genotype D had a higher prevalence of the G1896A mutation and the presence of a thymine at position 1858. Genotype A was associated with a higher ...

Naturally Occurring Mutations of Hepatitis B virus and Hepatitis C Virus in Korean Chronic Patients by Distinct CD4 T Cell Responses

Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are among the most common causes of chronic liver disease worldwide. The host immune pressure against hepatitis viruses during the chronic infection has led to mutations in their coding genes, which could play a pivotal role in the clinical outcomes of chronic patients. Our recent molecular epidemiologic studies regarding the HBV precore/core (preC/C) regions and HCV nonstructural 5B (NS5B) protein suggest the presence of distinct CD4 T cell immune pressure against HBV and HCV in Korean chronic patients. However, induced HBV and HCV mutations seem to exert an opposite effect on Korean chronic hepatitis B (CHB) and chronic hepatitis C (CHC) patients, respectively. On the basis of two of our recent papers, we focused in this review on the relationships between the mutation patterns of HBV preC/C and HCV NS5B, which were presumed to be caused by distinct CD4 T cell pressure in the Korean population and their effect on the clinical outcomes and liver disease progression of CHB and CHC patients.

Hepatitis B virus subgenotype C2- and B2-associated mutation patterns may be responsible for liver cirrhosis and hepatocellular carcinoma, respectively

The objective of this study was to examine hepatitis B virus (HBV) subgenotypes and mutations in enhancer II, basal core promoter, and precore regions of HBV in relation to risks of liver cirrhosis (LC) and hepatocellular carcinoma (HCC) in Southeast China. A case-control study was performed, including chronic hepatitis B (CHB; n=125), LC (n=120), and HCC (n=136). HBV was genotyped by multiplex polymerase chain reaction and subgenotyped by restriction fragment length polymorphism. HBV mutations were measured by DNA sequencing. HBV genotype C (68.2%) predominated and genotype B (30.2%) was the second most common. Of these, C2 (67.5%) was the most prevalent subgenotype, and B2 (30.2%) ranked second. Thirteen mutations with a frequency >5% were detected. Seven mutation patterns (C1653T, G1719T, G1730C, T1753C, A1762T, G1764A, and G1799C) were associated with C2, and four patterns (C1810T, A1846T, G1862T, and G1896A) were associated with B2. Six patterns (C1653T, G1730C, T1753C, A1762T, G1764A, and G1799C) were obviously associated with LC, and 10 patterns (C1653T, G1730C, T1753C, A1762T, G1764A, G1799C, C1810T, A1846T, G1862T, and G1896A) were significantly associated with HCC compared with CHB. Four patterns (C1810T, A1846T, G1862T, and G1896A) were significantly associated with HCC compared with LC. Multivariate regression analyses showed that HBV subgenotype C2 and C2-associated mutation patterns (C1653T, T1753C, A1762T, and G1764A) were independent risk factors for LC when CHB was the control, and that B2-associated mutation patterns (C1810T, A1846T, G1862T, and G1896A) were independent risk factors for HCC when LC was the control.

Hepatitis B Precore Protein: Pathogenic Potential and Therapeutic Promise

Hepatitis B virus (HBV), a small and economically packaged double-stranded DNA virus, represents an enormous global health care burden. In spite of an effective vaccine, HBV is endemic in many countries. Chronic hepatitis B (CHB) results in the development of significant clinical outcomes such as liver disease and hepatocellular carcinoma (HCC), which are associated with high mortality rates. HBV is a non-cytopathic virus, with the host's immune response responsible for the associated liver damage. Indeed, HBV appears to be a master of manipulating and modulating the immune response to achieve persistent and chronic infection. The HBV precore protein or hepatitis B e antigen (HBeAg) is a key viral protein involved in these processes, for instance though the down-regulation of the innate immune response. The development of new therapies that target viral proteins, such as HBeAg, which regulates of the immune system, may offer a new wave of potential therapeutics to circumvent progression to CHB and liver disease.

Mutations in Hepatitis B Virus Precore, Core Promoter, and "a" Determinant in Children with Chronic Hepatitis B Virus Infection / 대한소아소화기영양학회지

PURPOSE: The aim of this study was to determine the prevalence, types of variants, and clinical significance of mutations in precore, core promoter, and "a" determinant mutations in children with chronic hepatitis B virus infection. METHODS: Thirty-one patients with chronic hepatitis B virus infection who visited Seoul National University Children's Hospital in Korea between 2004 and 2005 were enrolled in this study. Serum HBV DNA was amplified by polymerase chain reaction (PCR) and the precore/core promoter and "a" determinant sequences were determined. RESULTS: Precore mutations were found in 11 of 27 patients (40.7%), and appeared more frequently in the HBeAg-negative group (p<0.05) compared to the HBeAg-positive group. G1896A was detected most frequently (81.8%). BCP mutations were found in 15 of 27 patients (55.6%) and the TA mutation (A1762T/G1764A) was the most common (86.7%). Mutations in the "a" determinant region were detected in 8 of 28 patients (28.6%), and amino acid changes were detected in 6 of 28 patients (21.4%). Of these mutations, substitutions at amino acid position 126 were found most frequently. CONCLUSION: In children with chronic hepatitis B virus infection, the most common mutations were G1896A in the precore region and TA mutation(A1762T/G1764A) in the core promoter region. Substitutions at amino acid position 126 was the most common mutation in the "a" determinant. Precore mutants were found to be significantly higher in HBeAg-negative patients. The high prevalence of mutations in the "a" determinant and low frequency of G145R were characteristic features. These mutations were not significantly associated with other clinical features except for high aminotransferase concentration in the core promoter variant group.

Prevalence of the precore G1896A mutation in Chinese patients with e antigen negative hepatitis B virus infection and its relationship to pre-S1 antigen

This study investigated the prevalence of the precore G1896A mutation in Chinese patients with hepatitis B e antigen (HBeAg) negative HBV infection and its relation to serum HBV pre-S1 antigen. The overall prevalence of the precore G1896A mutation was 72.6 percent in HBeAg-negative Chinese patients with detectable serum HBV DNA. The prevalence of the precore G1896A is significantly higher in Chinese HBeAg-negative patients with chronic hepatitis B than that in inactive HBV carriers with detectable serum HBV DNA. Serum pre-S1 and the precore G1896A mutation were simultaneously detected in most of Chinese HBeAg-negative patients.

Distribution of hepatitis B virus genotypes and viral load levels in Brazilian chronically infected patients in São Paulo city / Distribuição dos genótipos do vírus da hepatite B e níveis de carga viral em pacientes brasileiros cronicamente infectados na cidade de São Paulo

The objective of the present study was to evaluate the serum viral load in chronically infected Hepatitis B virus (HBV) patients and to investigate the distribution of HBV genotypes in São Paulo city. Quantitative HBV-DNA assays and HBV genotyping have gained importance for predicting HBV disease progression, have been employed for assessing infectivity, for treatment monitoring and for detecting the emergence of drug resistance. Twenty-nine Brazilian patients with suspected chronic hepatitis B were studied, using real time PCR for viral load determination and direct DNA sequencing for the genotyping. The serology revealed chronic HBV infection in 22 samples. The HBV-DNA was positive in 68 percent samples (15/22). The phylogenetic analysis disclosed that eleven patients were infected with HBV genotype A, two with genotype F and two with genotype D. Thus, the genotype A was the most prevalent in our study.

O objetivo do presente estudo foi avaliar a carga viral no soro de pacientes cronicamente infectados pelo vírus da Hepatite B (HBV) e investigar a distribuição de genótipos HBV na cidade de São Paulo. PCR quantitativo do HBV e genotipagem ganharam importância para a previsão de progressão da doença, empregada para avaliar a infectividade, para tratamento e acompanhamento e para detectar o aparecimento de resistência aos anti-retrovirais. Vinte e nove pacientes brasileiros com suspeita de hepatite B crônica foram estudados, utilizando PCR em tempo real para a determinação da carga viral e seqüenciamento direto para determinação do genótipo. A sorologia revelou que 22 estavam, de fato, cronicamente infectados pelo HBV. O HBV-DNA foi positivo em 68 por cento das amostras (15/22). Em sete casos, HBV-DNA foi indetectável por PCR quantitativo. A análise filogenética mostra que onze pacientes foram infectados com hepatite B genótipo A, dois com genótipo F e dois com genótipo D. Desta forma, o genótipo A foi o mais prevalente em nosso estudo.

Papel del genotipo y variantes precore/core del virus de la hepatitis B en el curso clínico y el tratamiento / Rol of Hepatitis B virus genotype and precore/core variants on antiviral treatment response and clinical outcome

Entre 500.000 y 1’000.000 de personas mueren anualmente en todo el mundo a causa de complicaciones relacionadas con la infección por el virus de la hepatitis B. Este virus presenta la tasa de mutación más alta entre los virus ADN que infectan al hombre. Aunque algunos de los genotipos del virus de la hepatitis B se han asociado al desarrollo de complicaciones durante la infección persistente, los estudios recientes sugieren un papel más importante a la aparición de ciertas variantes de este virus durante la infección. Una de las regiones del genoma del virus de la hepatitis B más analizadas y que presentan mayor frecuencia de mutaciones, es la regiónprecore/core y el promotor correspondiente; la doble mutación (A1762T/G1764A) del promotor es la de mayor impacto sobre la replicación del virus de la hepatitis B y la respuesta a la terapia antiviral. En esta revisión se muestra el papel del genotipo y las variantes de la región precore/core del virus de la hepatitis B en el contexto del curso clínico y el tratamiento antiviral.

The impact of antepartum interruption of intrauterine infection on the mutations of precore and core promoter regions of hepatitis B virus / 中华传染病杂志

Objective To investigate the impact of injecting hepatitis B immune globulin(HBIG)at third trimester of pregnancy on the nucleotide sequences of precore and basal core promoter(BCP)regions of hepatitis B virus(HBV)DNA.Methods One hundred and twenty pregnant women(67 in HBIG group and 53 in no-HBIG group)were enrolled in this study.Serum HBV DNA level was determined using quantitative real-time polymerase chain reaction(RT-PCR).Relevant serum markers (HBeAg,HBsAg)of HBV were detected by enzyme-linked immunosorbent assay(ELISA).Nucleotide fragments of HBV precore and BCP regions were amplified by nested PCR and then sequenced by automated DNA sequencer.Data were analyzed using t test and chi-square test.Results Sera of 33 women in HBIG group were collected before interruption with HBIG and at delivery.Precore and BCP regions of HBV DNA were amplified and sequenced successfully from double sera of 23 among 33 women. The rates of total nucleotide substitute in precore and BCP regions, that in precore region, and that in BCP region before and after interruption were 1.5% and 1.4%, 0.7% and 0.6%, 1.7% and 1.7%, respectively (Fisher's exact test, X2 =0.627, 0.689, 1.000, respectively,all P＞0.05). The rates of total mutations of hot points including 1896G→A,1899G→A,1762A→T,1764G→A before and after interruption were 27.2% and 13.0%, respectively (x2=5.717, P=0. 017). But the prevalences of these hot points mutations before and after interruption were 30.4%and 17.4%, 17.40/00 and 4.3%, 26.1% and 13.0%, 34.80/00 and 17.4%, respectively, which were all not significantly different (P＞0.05). The rates of nucleotide substitute in precore and BCP regions,that in precore region, and that in BCP region of 53 women in HBIG group and 47 women in no-HBIG group at delivery were 0.9% and 0.8%, 0.3% and 0.3%, 1.1% and 0.9%, respectively (Fisher's exact test, )x2=0.434, 0.839, 0.340, respectively, all P＞0. 05). The rates of total mutations of hot points of women in HBIG group and those in no-HBIG group at delivery were 5.7% and 10.1%,respectively, which was not significantly different (P＞0.05). These hot points mutations including 1896G→A,1899G→A,1762A→T, 1764G→A of women in HBIG group and those in no-HBIG group at delivery were 9.4% and 14.9%, 0 and 2. 1%, 7.5%0 and 10.6%, 5.7% and 12.8%, respectively,which were all not significantly different ( P＞0.05). Conclusions Antepartum interruption of HBV intrauterine infection with HBIG may not raise the nucleotide mutations in precore and BCP regions of HBV DNA. On the other hand, antepartum interruption may decrease mutations of hot points in the precore and BCP regions of HBV DNA.

Hepatitis B virus genotype E detected in Brazil in an African patient who is a frequent traveler

Genotype E of hepatitis B virus (HBV) has not been described in Brazil and is found mainly in Africa. Genotype A is the most prevalent in Brazil, and genotypes B, C, D, and F have already been reported. We report here an HBV genotype E-infected patient and some characterization of surface (S) protein, DNA polymerase (P) and precore/core (preC/C) coding regions based on the viral genome. The patient is a 31-year-old black man with chronic hepatitis B who was born and raised in Angola. He has been followed by a hepatologist in São Paulo, Brazil, since November 2003, and he is a frequent traveler to Latin America, Africa, and Europe. In 2003, he was diagnosed with HBV infection and started treatment with lamivudine with the later addition of adefovir dipivoxil. No known risk factor was identified. Serologically, he is HBsAg and anti-HBe positive, but HBeAg and anti-HBs negative. DNA sequence analysis of the S/P region confirmed that this patient is infected with genotype E, subtype ayw4. The preC/C region showed G1896A and G1899A mutations but no mutations in the basal core promoter. Nucleotide substitutions common in genotype E were also observed (C1772, T1858 and A1757). Although this is not an autochthonous case and there is no evidence of further spread, the description of this case in Brazil highlights the current risk of viral genotypes spreading with unprecedented speed due to constant travel around the world.

Hepatitis B virus genotypes and mutations affecting HBeAg production: A Malaysian perspective

Infection by hepatitis B virus (HBV) is a major global health-care problem. HBV is an accepted factor in the elevated risks for liver disease such as cirrhosis and development of hepatocellular carcinoma. This problem is particularly prevalent in the Asia-Pacific region which includes Malaysia. During infection, the hepatitis B e antigen (HBeAg) is produced in the hosts. This antigen is an important serological marker for diagnosing chronic hepatitis B. Seroconversion to anti-body (anti-HBe) corresponds to the improvement of disease prognosis. However, certain mutations such as the core promoter dual mutations (A1762G1764→T1762A1764), the codon 15 variants (C1858/ T1858) and the precore stop codon mutations (TGG→TAG) can affect the HBeAg expression. This has diagnostic and clinical implications. Besides that, the HBV can be grouped into eight genotypes (A to H). Moreover, genotypic subtypes and recombinants have been observed as well. Studies have observed that these can differ in their affiliations with the mutations above as well as with disease prognosis.

Clinical study on YMDD mutation and pre-core C region and core promotor region mutation of the patients with chronic hepatitis B virus receiving lamivudine therapy / 中国基层医药

Objective To mvesugate the Tate of YMDD mutation accompamed with pre-core(region and core promotor region mutation and the clinical significance.Methods YMDD mutation and pre-core(at 1896 nu- cleotide)region and core promotor region(at 1762.1764 nucleotide)mutation were detected from the 122 patients with chronic hepatitis B virus after receiving lamivudine treatment above 6 months.Results 40 cases were tested for YMDI)mutations in 122 HBV patients with lamivudine treatment,and the positive rate of YMDD mutation was 32.8 %.After YMDD mutation,ALT,AST and HBV DNA of the patients significantly increased(P0.05).Conclusion The patients with YMDD mutation had higher rate of pre-core region(at 1896 nucleotide)and basal core promotor region(at 1762, 1764 nucleotide)mutation than those without YMDD mutation,but there was no correlation between the mutation and the deterioration of disease condition and the bad prognosis.

Prevalência de genótipos e de mutantes pré-core A-1896 do vírus da hepatite B e suas implicações na hepatite crônica, em uma população da Amazônia oriental / Prevalence of hepatitis B virus genotypes and the occurrence of precore mutation A-1896 and to correlate them with the clinical presentation of chronic hepatitis, in a population group of the eastern Amazon region

A infecção pelo virus da hepatite B apresenta amplo espectro de manifestações clínicas. Objetivando conhecer os genótipos do HBV mais prevalentes e determinar a ocorrência da mutação pré-core A-1896, em uma população da Amazônia oriental, correlacionando com o diagnóstico clínico, foram selecionados 51 pacientes portadores crônicos de HBsAg e HBV-DNA positivos e divididos em três grupos: grupo A (n=14, pacientes assintomáticos); grupo B (n=20, sintomáticos HBeAg positivos) e grupo C (n=17, sintomáticos HBeAg negativos), sendo usado o sequenciador automático ABI modelo 377 para identificação de genótipos e mutantes pré-core. Os resultados evidenciaram o genótipo A como o mais prevalente, 81,8%, 89,5% e 93,7%, nos grupos A, B e C, respectivamente. A mutação pré-core A-1896 foi encontrada em 11,5% (3/26), sendo todos assintomáticos. Concluiu-se que na população estudada o genótipo A foi o mais prevalente e houve baixa ocorrência do mutante pré-core A-1896, ambos não se constituindo fatores agravantes da doença hepática.

Hepatitis B virus (HBV) infection presents itself with a variety of clinical manifestations. The present work aims to describe the prevalence of HBV genotypes and the occurrence of precore mutation A-1896 in a population group of the Eastern Amazon region of Brazil and to correlate them with the clinical presentation of chronic HBV infection. 51 HBsAg carriers (HBV-DNA positive) were selected and divided into three groups: A (14 asymptomatic subjects), B (20 HBeAg positive symptomatic patients) and C (17 HBeAg negative symptomatic patients). Using an automa ed DNA sequencer ABI model 377 by sequencing for determined of genotypes and precore mutation. The results showed that the genotype A was the most commonly found (81,1%, 89,5% and 93,7% in groups A, B and C, respectively) and precore mutation A-1896 was described in 11,5% (3/26) of group A subjects. Genotype A of HBV was the most prevalent (89,1%) and low occurrence of precore mutation A-1896, both not associate with the worst outcome of the chronic infection of HBV.

The Prevalence and Clinical Significance of Precore and Core Promoter Mutations in Korean Patients with hronic Hepatitis B Virus Infection / 대한간학회지

BACKGROUND/AIMS: Precore and core promoter mutations of hepatitis B virus (HBV) have been reported in Korea but their prevalence and clinical significance have not been determined. The aims of this study were to determine the prevalence of precore and core promoter mutations and their relationships to hepatitis B e antigen (HBeAg) status, viral replication level, and severity of liver disease in Korea. METHODS: Among the patients who visited the Liver Diseases Clinics (Chung Ang University Hospital) between December 1998 and August 1999, 150 patients were randomly selected: 50 HBeAg-positive HBV-DNA positive patients by a branched DNA (bDNA) assay, 50 HBeAg-negative bDNA-positive patients, and 50 HBeAg-negative bDNA-negative patients. Serum HBV-DNA was amplified by a polymerase chain reaction (PCR) in these patients and the core promoter/precore HBV sequence was determined in 135 of the patients whose sera were positive for HBV-DNA by PCR. RESULTS: All of the 135 determined HBV-DNA sequences had HBV genotype with T at nucleotide 1858. Precore mutation (A1896) was detected in 95.7% of HBeAg-negative bDNA-positive patients and 94.9% of HBeAg-negative bDNA-negative patients. In HBeAg-positive patients 88% had wild type and 12% had mixture of wild type and A1896 mutant. Core promoter TA mutation (T1762/A1764) was detected in 93.5% of HBeAg-negative bDNA-positive patients, 94.9% of HBeAg-negative bDNA-negative patients and 74% of HBeAg-positive patients. No correlation was found between the presence of precore/core promoter mutations and liver disease severity or HBV-DNA levels. CONCLUSION: Precore stop codon mutation occurred almost invariably, along with HBeAg seroconversion, irrespective of subsequent viral replication levels or disease severity. Core promoter TA mutation was frequent both in the HBeAg-positive patients and HBeAg-negative patients irrespective of viral replication levels or disease severity.

The association of hepatitis B virus precore/basic core promoter mutations with genotype and progression of liver disease / 中华消化杂志

Objective To study association of hepatitis B virus(HBV) precore (pre c)/basic core promoter(BCP) mutations with the genotype or the progression of liver disease. Methods The serum samples from 148 patients with HBV-relative diseases were collected, including 31 asymptomatic carriers, 32 with chronic hepatitis B (CHB), 40 with liver cirrhosis(LC) and 45 with hepatocellular carcinoma(HCC). The genes covering HBV pre c and BCP were amplified by nested polymerase chain reaction (nPCR). The PCR products were subjected to direct sequencing and the mutations in pre c 1896 and BCP 1762/1764 were determined by sequence analysis. HBV genotypes were also detected in the sera by restriction fragment length polymorphism based on S-gene PCR products. Results Of 148 serum samples of HBV, 128 were successfully genotyped and sequenced. There were 60 genotype B and 68 genotype C. The mutation in pre c (A1896) was significantly higher in genotype B than in genotype C (48.3% vs 29.34%, P

The influence of hepatitis B virus basic core promoter/precore mutations on adv ance of liver disease / 中国实用内科杂志

Objective To investigate the relationship between hepatitis B virus (HBV) basic core promoter(BCP)/precore(PreC) mutations and severity of liver dis ease. Methods In 113 patients chronically infected with HBV, do uble mutations in BCP(T1762/A1764) and PreC mutation(A1896) were determined by INNO-LiPA and HB V genotype was determined by S gene sequencing. Results Whether in all patients or in patients infected by single genotype C, compared with AsC, the prevalence of double mutations in BCP(T1762/A1764) was higher in patients with CHB, LC and HCC[(24.1% vs 2.8%,? 2=5.93, P0.05). Conclusions The doubl e mutations in BCP(T1762/A1764 ) may be related to progressive liver disease in patients with chronic HBV infec tion.

Mutations in Hepatitis B Virus Precore and Core Promotor in Children with Chronic Hepatitis B Infection - Comparison Between Vertical and Non-vertical Transmission

PURPOSE: The aims of this study were to investigate the frequencies and role of hepatitis B virus(HBV) precore and core promotor mutations in children with chronic hepatitis B infection. METHODS: Sera from 46 children with chronic hepatitis B infection were analyzed by direct sequencing of polymerase chain reaction product of HBV DNA. In this study, the patients were divided into vertical and non-vertical groups according to the mode of HBV transmission. Statistical analysis was performed by using Fisher's exact test. RESULTS: Forty-six adr type of HBV DNA were analyzed. The mutations in HBV precore region were observed in 12(26.1Yo) of 46 cases. The GA mutation of nucletide(nt) 1896 was observed in 5 cases(10.9Yo). The frequency of mutations in HBV precore region of the non-vertical group (6/16; 37.5Fo) was higher than that of the vertical group(6/30; 20M), but there was no statistical significance. The mutation in HBV core promotor region was observed in 40(87.0%) of 46 cases. The A-->T mutation of nt 1762 or G-->A mutation of nt 1764 were observed in 24(52.2%) of 46 cases, and 23 cases revealed combined mutation at both positions 1762 and 1764. The frequency of mutations in HBV core promotor region of the vertical group(28/30; 93.3Yo) was higher than that of the non-vertical group(12/16; 75.0M), but there was no statistical significance. The frequencies of mutations in HBV precore and core promotor regions of the HBeAg negative patients was higher than that of HBeAg positive patients, but there was no statistical significance. Also there were no significant correlations between the frequencies of mutations in HBV precore and core promotor regions and AST, ALT level or the level of HBV DNA. CONCLUSION: These observations suggest that mutations in HBV precore and core promotor regions were frequently detected in children with chronic hepatitis B infection. There were no statistical significant differences in the frequencies of mutations in HBV precore and core promotor regions between vertical and non-vertical transmission groups. (J Korean Pediatr Soc 2000; 43:779-791)

Clinicopathologic Significance of HBV X, Core promoter, Precore Mutants in Patients with Chronic HBV Infection / 대한간학회지

BACKGROUND/AIMS: There have been much debates on the effects of HBV mutants on the clinical course of HBV-associated chronic liver diseases. The purpose of this study was to define the relationship among HBV mutants, severity of hepatitis and expression patterns of HBcAg METHODS: HBV DNA was extracted from the liver tissue of 31 patients who had been HBsAg positive for more than 6 months. The amplification of X was performed as well as core promoter/precore region of HBV DNA by polymerase chain reaction and then direct sequencing of them. Pathologic severity was classified utilizing Scheuer's scoring system and immunohistochemical staing for HBcAg in hepatocytes was performed. The expression patterns of HBcAg were divided into four types according to expression location of HBcAg: Type I as a nuclear predominant expression of HBcAg; Type II as mixed patterns, combined expression of cytoplasmic and nuclear localization of HBcAg; Type III as a diffuse cytoplasmic expression of HBcAg; and Type II as an inclusive body-like expression in cytoplasm. RESULTS: In investigating the relationship between HBV mutants and clinical findings, ALT, HBV DNA and hepatitis activity index (HAI) in hepatitis with wild HBV were normal to high but those in hepatitis with core promoter or precore mutants were high . There were no statistically significant differences (p=0.062). In terms of the relationship between HBV mutants and the expression pattern of HBcAg, type I, II, IV were noticed in hepatitis with wild HBV but in almost all mutants cases type III, II were noticed (p<0.01). The score of HAI increased as the number of the expression pattern of HBcAg increased from type I to type III or IV(p<0.05). No relationships among the mutation in X region, the mutations in other regions and clinicopathological severity could be found. CONCLUSION: The mutation in X, core promoter and precore region had little association with the severity of hepatitis. And the relationship did not exist between precore mutants and X mutants. The expression pattern of HBcAg could be a useful indicator in determining what stage of chronic hepatitis B is in and whether mutant strains exist.

Hepatitis B Virus DNA Mutation, Pattern of Major Histocompatibility Class-I among Familial Clustered HBV Carriers in Relation to Disease Progression / 예방의학회지

OBJECTIVES: Chronic HBsAg carriers are the principal source of infection for other susceptible people, and are themselves at high risk of developing serious liver diseases. In Korea, it has been estimated that 65-75% of the HBsAg positives remained as persistent carriers. Additionally, familial clustering of HBV infection has frequently been observed among carriers. Some would become progressive, chronic hepatitis patients, and others would not. The aim of this study was to evaluate the association between various factors, such as the duration of infection, type of virus, mutation of precore/core region in HBV, major histocompatibility class-I, and developing chronic liver diseases among familial HBV carriers. METHODS: Chronic carrier status was identified by repeated serological tests for HBsAg at intervals of six months or more. A familial chronic carrier was defined when the disease was observed in a family member over two generations. Two families were recruited, among which a total of 20 chronic HBsAg carriers(11 carriers in No.1, and 9 in No.2 family) were identified. Data on the general characteristics and liver disease status were collected. Identification of the HBV-DNA was successful only for 13 subjects among the 20 carriers. Analysis of viral DNA in terms of subtype, pre-core and core region mutations was carried out. The type of major histocompatibility class-I for the 13 subjects was also analysed. RESULTS & CONCLUSIONS: Seven of 10 chronic HBV carriers of the 1st generation and one of 10 of the 2nd generation were clinical patients with chronic hepatitis, the others, three of the 1st and nine of the 2nd generation, were asymptomatic carriers. This data indicates that the duration of HBV carriage is one of the major factors for disease severity. The subtype of HBsAg analysed using HBV-DNA identified in 13 carriers were adr, and the pattern of precore nonsense mutation in HBV-DNA was identical among family members, which means that the same virus strains were transmitted between the family members. The association between the precore or core mutations in HBV-DNA and the disease severity was not observed. While it was suggested that a specific type of MHC class-I may be related to disease progression.