Significance of the expression of pre-S proteins in mononuclear blood cells in chronic hepatitis caused by the hepatitis B virus

In chronic viral type B hepatitis, the presence in the serum of pre-S proteins of hepatitis B virus (HBV) envelope reflects viral replication. As peripheral blood mononuclear cells (PBMC) are known to be target cells for HBV replication, the aim of our study was to investigate the clinical relevance of pre-S protein expression in PBMC. Fifty-seven patients with chronic type B hepatitis and HBs antigenemia were studied. Following separation using the Ficoll gradient, the PBMC were lysed and studied for pre-S proteins by Western blot. HBs Ag and HBc/e Ag were assayed in parallel by radioimmunoassay. HBs Ag was detected in PBMC in 86 percent of cases, HBc/e Ag in 28 percent of cases and pre-S proteins in 34 percent of cases. A statistically significant association was found between the presence of HBc/e Ag in PBMC and both the serum HBe Ag (chi 2 test, p less than 0.01) and the serum viral DNA/DNA polymerase (t test, p = 2.10(-4)). The pre-S protein expression in PBMC was significantly associated with higher levels of DNA/DNA polymerase activity (chi 2 test, p less than 0.05). The expression of pre-S proteins in PBMC appears therefore to correlate with the HBV viral replication phase. The HBc/e Ag and pre-S protein detection in PBMC therefore offers a reliable non invasive approach to tissular viral replication. The clinical relevance of pre-S testing in PBMC was illustrated by the study of 12 cases of chronic active hepatitis positive for anti-HBe but with no or low level of serum DNA polymerase activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum titers of pre-S(2) antigen in patients with acute and chronic type B hepatitis: relation to serum aminotransferase activity and other hepatitis B virus markers

The peptide which is encoded by the pre-S(2) region of hepatitis B virus DNA, the pre-S(2) antigen, was determined quantitatively by an enzyme immunoassay system employing monoclonal antibodies. The prevalence and titer of pre-S(2)Ag were 91.9% (91/99) and 10,356 +/- 19,053 units (mean +/- S.D., arbitrary units) for hepatitis B e antigen (HBeAg)-positive patients with acute and chronic HBV infection and 86.0% (74/86) and 952 +/- 1,565 units for HBeAg-negative subjects. In four patients with acute hepatitis B, pre-S(2)Ag titers changed in parallel with HBV DNA levels, and the disappearance of pre-S(2)Ag from serum was associated with a rapid fall of ALT levels into the normal range, whereas the fluctuation of pre-S(2)Ag titer correlated with persistence of ALT elevations. In all of the 19 episodes of acute exacerbation of hepatitis which occurred in nine patients with chronic active hepatitis B, a significant elevation of pre-S(2)Ag titer was observed, closely overlapping an increase or appearance of HBV DNA, and its peak preceded peaks of ALT by 1 to 11 weeks (mean +/- S.D. = 4.26 +/- 2.57 weeks). These observations suggest that quantitative measurement of pre-S(2)Ag would be useful for estimation of the magnitude of HBV replication and would help predict the prognosis of acute hepatitis B and of acute exacerbation in chronic hepatitis B.     

Hbc ag, anti-HBC, and DNA polymerase activity in transfused recipients followed prospectively.

Hepatitis B core antigen, antibody to core antigen, and DNA polymerase activity were measured in sera from a select group of post-transfusion hepatitis B patients who had been followed prospectively following blood transfusion. Preliminary results of this study have revealed (1) that RIA testing of blood would not eliminate but would reduce post-transfusion hepatitis B infections by about 50 per cent; (2) that infection with HB virus is modified or aborted in the presence of pre-existing antibody to HB surface antigen; and (3) that transfusion of blood containing anti-HBs does not increase the risk of post-transfusion hepatitis B. HBc Ag and/or DNA polymerase activity were observed in the sera of all recipients tested who developed liver enzyme abnormalities along HBs Ag and anti-HBc. DNA polymerase activity usually occurred in the early stages of incubation before the transaminase became abnormal, whereas HBc Ag was more often associated with increasing enzymatic evidence of liver damage, suggesting release of core structures from the hepatocytes. The presence of DNA polymerase without detectable HBc Ag may be due to the presence of intact Dane particles in the sera, preventing recognition of the core antigen. No serological evidence of hepatitis B was observed in the sera of 24 other recipients who developed abnormal transaminases. Immunoelectron microscopy of these same sera revealed evidence of exposure to hepatitis A antigen following transfusion in at least two recipients.     

Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA.

The hepatoblastoma cell line Hep G2 was transfected with a plasmid carrying the gene that confers resistance to G418 and four 5'-3' tandem copies of the hepatitis B virus (HBV) genome positioned such that two dimers of the genomic DNA are 3'-3' with respect to one another. Cells of one clone that grew in the presence of G418 produce high levels of hepatitis B e antigen and of hepatitis B surface antigen. HBV DNA is carried by these cells as chromosomally integrated sequences and episomally as relaxed circular, covalently closed, and incomplete copies of the HBV genome. Viral DNA was detected also in conditioned growth medium at the buoyant densities characteristic for infectious Dane and immature core particles. Finally, HBV-specific components morphologically identical to the 22-nm spherical and filamentous hepatitis B surface antigen particles as well as 42-nm Dane particles were visualized by immunoelectron microscopic analysis. Therefore, we have demonstrated that the Hep G2 cell line can support the assembly and secretion not only of several of the replicative intermediates of HBV DNA but also of Dane-like particles. This in vitro system can now be used to study the life cycle of HBV and the reaction of immunocompetent cells with cells carrying HBV.     

Pathogenesis of chronic liver disease in patients with chronic hepatitis B virus infection without serum HBeAg

Chronic hepatitis B in patients lacking hepatitis B e antigen has been attributed to a hepatitis B virus variant (G-to-A mutation at nucleotide 1896 in the precore region of the genome). We therefore assessed the frequency and significance of this variant among 43 United States patients (10 with chronic hepatitis B seropositive for e antigen, 19 seronegative for e antigen, and 14 healthy carriers). Sera were tested for HBV DNA by polymerase chain reaction and branched DNA assay. The A₁₈₉₆ variant was detected by direct sequencing and ligase chain reaction. Serum HBV DNA was more frequently found among patients with e antigenpositive than e antigen-negative chronic hepatitis B. Viral titers were generally higher in those with e antigen. None of the e antigen-positive and only 24% of e antigen-negative patients harbored the A₁₈₉₆ variant. Patients infected with the variant were more often Asian, had had hepatitis B for longer and had higher levels of viral DNA than HBeAg-negative patients with the wild-type virus. The A₁₈₉₆ variant was found exclusively in patients infected with HBV genotypes C and D. Thus, the A₁₈₉₆ variant is uncommon in the United States. The activity of liver disease appears to be more closely related to the level of HBV replication than the presence of mutations at nucleotide 1896 in the genome.     

Discordant e antigen, DNA polymerase activity, and Dane particle responses in two patients representing an index case-contact case pair with hepatitis B virus infection.

Discordant results for e antigen, DNA polymerase activity, and Dane particle frequency were noted in the sera of two patients representing an index case-contact case pair with chronic aggressive hepatitis B. The lack of correlation between presence of e antigen and the infecting viral strain, as well as the strong relationship of e antigen to persistent and significant viremia, emphasize the role of the host's immune response in the expression of this antigen.     

Significance of hepatitis B viremia levels determined by a quantitative polymerase chain reaction assay in patients with hepatitis B e antigen-negative chronic hepatitis B virus infection

OBJECTIVES: In hepatitis B e antigen (HBeAg)-negative chronic hepatitis B virus (HBV) infection, the clinical relevance of low viremia levels remains unclear. We evaluated the clinical significance of a single baseline serum HBV DNA measurement by a quantitative polymerase chain reaction (PCR) assay in this setting. METHODS: In total, 196 patients with HBeAg-negative chronic HBV infection (62 inactive carriers, 134 with chronic hepatitis B) were studied. ALT activity was normal at baseline in 25/134 HBeAg-negative chronic hepatitis B patients (18.7%), whereas it remained normal throughout follow-up in all inactive carriers. RESULTS: HBV DNA was <30,000 copies/ml in 14 (10.5%) and <100,000 copies/ml in 17 (12.9%) HBeAg-negative chronic hepatitis B patients, whereas it was <30,000 copies/ml in all inactive carriers (undetectable in 14). In particular, HBV DNA levels were <100,000 copies/ml in eight (32%) and <30,000 copies/ml in five (20%) of the 25 patients with HBeAg-negative chronic hepatitis B and normal baseline ALT values. HBV DNA levels with a cut-off at 30,000 or 100,000 copies/ml could correctly classify 92.9% or 91.3% of patients with HBeAg-negative chronic HBV infection, whereas ALT or IgM anti-HBc (IgM class antibody to HBV core antigen) index > 0.200 could correctly classify only 87.2% and 82.1% of patients, respectively. A combined HBV DNA and IgM anti-HBc index performed better by correctly classifying 94.4% of cases. CONCLUSIONS: Serum HBV DNA levels evaluated by sensitive quantitative PCR assays can be used for differentiation between HBeAg-negative chronic hepatitis B and inactive hepatitis B surface antigen carrier state, but the cut-off level should be set at approximately 30,000 copies/ml and certainly lower than the recently suggested level of 100,000 copies/ml.     

慢性乙型肝炎患者血清前S1抗原检测的意义

目的探讨PreS1抗原检测的临床价值。方法采用酶联免疫吸附试验法检测421例慢性乙型肝炎患者血清PreS1抗原和HBV标记物;采用荧光定量PCR法检测HBVDNA。结果在421例慢性乙型肝炎患者中,HBVDNA阳性者367例,其中PreS1Ag阳性者188例(51.2%),HBeAg阳性者119例(32.4%),后两者有显著性差异(P0.01);在高HBVDNA载量(105～107copies/ml和107copies/ml)组患者中,PreS1Ag阳性率(60.2%,60.0%)显著高于HBVDNA阴性组(33.3%)和低载量(103～105copies/ml)组(41.9%,P0.01);但在421例患者中,PreS1Ag阳性率(48.9%)低于HBVDNA(87.2%,P0.01)。结论 PreS1Ag能够较HBeAg更好地反映HBV在体内的复制状态,但尚不能代替HBVDNA的检测。     

Immunologic mechanisms in hepatitis B assayed by antigen-binding lymphocytes.

Immune mechanisms in hepatitis B virus (HBV) infection were investigated in 16 persons with and without hepatitis using tests for HBS Ag, anti-HBS, anti-HBC and 125I-labeled HBS Ag binding lymphocytes (ABL) in peripheral blood. Anti-HBC, which is an evidence of a recent or current HBV infection, was detected in all HBS Ag positive sera. High counts of ABL correlated with the presence of anti-HBS in serum but not with anti-HBC or with HBS Ag. In patients with chronic hepatitis, and in asymptomatic carriers of HBS Ag, there was a trend towards low counts of ABL, which may represent partial tolerance ot HBS Ag in carriers of this particle. Further work on ABL for HBS Ag and HBC Ag should enhance our understnading of immunologic responses to the antigens of the hepatitis B virus.     

Hepatitis B core and surface antigen expression in HBeAg and HBV DNA positive chronic hepatitis B: correlation with clinical and histological parameters

The interrelationship between hepatitis B virus (HBV) infection, hepatic injury and clinical activity in chronic HBV infection is incompletely understood. We have scored histologic activity, the expression of hepatitis B core (HBcAg) and hepatitis B surface antigen (HBsAg) and assessed HBV replication to correlate HBV antigen expression with histologic disease. Forty-seven formalin-fixed, percutaneous liver biopsies from HBeAg carriers were studied. Twenty-nine were Black, 16 Caucasian and two Oriental. Fifty-nine percent had chronic active, 35% chronic persistent hepatitis and 14% cirrhosis. None were positive for antibodies to Human Immunodeficiency Virus (HIV). HBsAg and HBcAg in tissue were detected by immunochemical staining. Diffuse HBsAg staining was observed in 10/15 patients with CPH, but there was no correlation between histologic score and HBsAg expression. Intracytoplasmic HBcAg was observed in patients seroconverting to anti-HBe, but was also detected in patients with minimal hepatitis. An inverse correlation between histologic score and HBcAg expression was observed. HBcAg expression was more widespread in patients with CPH (mean 37%) than in CAH (mean 18%). A positive correlation was observed between serum aminotransferase concentrations and histologic score. Although no consistent pattern can be discerned, HBcAg expression and hepatic injury are frequently dissociated in patients with chronic HBV infection; complex host responses may determine the variable degree of disease activity and hepatic injury.     

Relationship of clinical and virological factors with hepatitis activity in hepatitis B e antigen-negative chronic hepatitis B virus-infected patients

To investigate the factors associated with active disease among hepatitis B surface antigen (HBsAg) positive/hepatitis B e antigen (HBeAg)-negative chronic hepatitis B virus (HBV) infection we studied chronic HBV infected patients who had undetectable HBeAg at the first visit. HBV DNA was determined by the cross-linking assay (NAXCOR) and polymerase chain reaction (PCR). Mutations in the core promoter and precore regions and viral genotypes were studied. Clinical outcome of these patients were followed and categorized as: (i) relapse (ALT > 200 IU/L or three times the previous levels); (ii) active hepatitis (elevated ALT < 200 IU/L with concomitant detectable HBV DNA); or (iii) remission. A total of eighty-five patients were followed up for 5.5 ± 1.0 years. At first visit, 31 (36.5%) patients had elevated ALT levels, 12 (14.1%) had measurable HBV DNA by the cross-linking assay and 26 (30.6%) by PCR. Sixteen (18.8%) patients had hepatitis relapse, 13 (15.3%) had active hepatitis, and 56 (65.9%) remained in remission. Core promoter and precore stop codon mutants were found in 27 and 12 patients, respectively. Eleven and 20 had genotype B and C HBV, respectively. Initial elevated ALT and detectable HBV DNA were associated with active liver disease. Patient demographics, viral mutants or genotypes failed to predict disease activity. Hence, serum ALT and HBV DNA levels offer the best prediction of natural course of HBeAg-negative chronic HBV infection.     

Treatment of chronic viral hepatitis with nitazoxanideand second generation thiazolides

Nitazoxanide, the first thiazolide, was originally developed for the treatment of Cryptosporidium parvum. More recently, antiviral activity of nitazoxanide against hepatitis B virus （HBV） and hepatitis C virus was recognized in in vitro systems. These basic studies led to phase Ⅱ clinical trials that demonstrated the safety and efficacy of nitazoxanide in combination with peginterferon, with or without ribavirin, in the treatment of chronic hepatitis C genotype 4. The sustained virologic response rate was 79% and 80% in two studies, which was higher than the response rate of 50% with the standard of care with peginterferon plus ribavirin. In very preliminary studies of patients with chronic hepatitis B, nitazoxanide suppressed serum HBV DNA and led to loss of hepatitis B e antigen in the majority of patients and hepatitis B surface antigen in approximately a quarter of patients. Randomized controlled studies of naive and nonresponder patients with chronic hepatitis C genotype 1 are underway, new second generation and controlled release thiazolides are being developed, and future studies of patients with chronic hepatitis B are planned.     

Clinical utility of a simple quantitative determination of hepatitis B e antigen

The titre of serum hepatitis B e antigen (HBeAg) was determined by using a simple quantitative system which is a slight modification of a commercially available enzyme immunoassay, and the relationship of serum HBeAg titre, hepatitis B virus (HBV)-associated DNA polymerase activity and HBV DNA level was evaluated. The HBeAg titre was found to have a significant correlation with DNA polymerase activity (r= 0.8153) and HBV DNA level (r= 0.8001). In 12 acute exacerbations of hepatitis in 10 patients with chronic type B hepatitis, these three parameters were found to be elevated either prior to or concurrent with the elevation of serum alanine aminotransferase, and the HBeAg titre peaked either simultaneously with or 1–4 weeks after the peaks of DNA polymerase activity and HBV DNA level, with average time lags of 1.00 weeks (s.d. = 1.29) and 1.08 weeks (s.d. = 1.25), respectively. Quantified HBeAg can be considered as a serum marker indicative of HBV replication, as is the case with DNA polymerase and HBV DNA. Furthermore, the HBeAg assay has the advantages of simplicity and low cost and it does not require special equipment. Therefore, quantification of HBeAg should be employed widely in clinical practice.     

Characteristics of hepatitis B X antigen, antibodies to X antigen, and antibodies to the viral polymerase during hepatitis B virus infection

The characteristics of hepatitis B virus (HBV) X antigen (HBxAg) and antibodies against the X antigen (anti-HBx) and the viral polymerase (anti-pol) were determined in 85 HBV-infected patients. HBxAg was detected in sera positive for HBV e antigen (HBeAg) and HBV DNA in patients with acute and chronic hepatitis, while anti-HBx appeared when markers of viral replication became undetectable. HBxAg was common in the liver among patients with chronic hepatitis independent of HBV replication markers but was closely correlated with elevated alanine aminotransferase, implying that HBxAg in liver may be important in the pathogenesis of chronic infection. Anti-pol was detected in many samples positive for HBeAg and HBV DNA and less often in serum samples without markers of HBV replication, suggesting that this marker could reflect ongoing viral replication in the liver, even though such markers were absent from sera.     

Hepatitis B virus DNA in Kaposi sarcoma.

DNA extracted from Kaposi sarcoma biopsies was analyzed for the presence of hepatitis B viral (HBV) DNA sequences by Southern blot hybridization. The probes used were (i) molecularly cloned subgenomic fragments of HBV DNA and (ii) HBV DNA extracted from Dane particles that included DNA sequences that encode hepatitis B surface antigen and core antigen. The limited study reported here demonstrates that HBV DNA exists mainly in an episomal state in the Kaposi sarcoma tissue and that these DNA forms are different from those found in hepatitis B virion. The presence of various DNA forms suggests that HBV DNA is engaged in DNA replication, which implies that the tissue is infected with HBV.     

乙型肝炎病毒表面抗原阳性患者前-S1抗原和前-S2抗原与HBV DNA和HBV-M相关性分析

目的检测乙型肝炎病毒患者乙型肝炎病毒前-S1抗原（HBV pre-S1-Ag）、前-S2抗原（HBV pre-S2-Ag）、乙型肝炎病毒DNA（HBV DNA）和乙型肝炎病毒E抗原（HBeAg）,探讨其相关性和临床应用价值。方法应用酶联免疫测定法（ELISA）分别检测HBV pre-S1-Ag、HBV pre-S2-Ag和乙型肝炎病毒标志物（HBV-M）,荧光定量聚合酶链反应法（FQ-PCR）检测HBV DNA,并对检测结果进行统计学分析。结果 HBsAg阳性者中,pre-S1-Ag、pre-S2-Ag、HBV DNA阳性者分别为594例、541例、629例,其阳性率分别为66.29%、60.38%、70.20%。HBeAg阳性组pre-S1-Ag、pre-S2-Ag、HBV DNA的阳性率分别为90.21%、74.46%、93.32%,显著高于HBeAg阴性组的45.28%、48.01%、49.89%,差异有统计学意义（P〈0.01）。随着HBV DNA载量的增高,pre-S1-Ag、pre-S2-Ag、HBeAg阳性率随之增高。结论 pre-S1-Ag、pre-S2-Ag与HBV DNA和HBeAg阳性检出率具有显著相关性。联合检测pre-S1-Ag、pre-S2-Ag、HBV DNA及HBV-M,有助于HBV早期诊断、疗效观察和预后判断。     

The effect of recombinant hepatitis B vaccine therapy in chronic hepatitis B infection.

BACKGROUND/AIMS: Immune-modulator and antiviral treatments for carriers of hepatitis B virus are known to have poor efficacy with a high cost and frequent side effects, which has led to investigation of new treatment modalities. The aim of this study was to determine the efficacy of recombinant hepatitis B vaccine in the treatment of patients with chronic hepatitis B (HBV DNA >5 pg/ml, ALT>60) infection diagnosed histopathologically and asymptomatic carriers (HBV DNA<5 pg/ml, ALT <40, Anti Hbe positive) of the virus. METHODS: The vaccine (Gen Hevac B Pasteur) was administered at baseline and at months one and six to patients with chronic hepatitis and asymptomatic carriers. Ten cases with chronic hepatitis B infection were assigned to a control group to whom no treatment was given. Biochemical and microbiological investigations were performed at baseline and at months three, six and twelve in all cases. Seroconversion of Hbe Ag, loss of HBV DNA and normalization of ALT were considered as a positive response. RESULTS: Patients with chronic hepatitis B who were given the vaccine were found to have significantly low levels of HBV DNA at 12 months (63.2+/-20pg/ml) compared to baseline values (174.4+/-36.9pg/ml), while controls were found to have high levels of HBV DNA at 12 months (223.1+/-33pg/ml) compared to baseline values (165.2+/-33.2pg/ml) (p<0.05). At 12 months, HBV DNA had become negative in seven of 19 patients given the vaccine (36.8%) Four patients with chronic hepatitis (36.35%) were observed to have HBeAg seroconversion and one patient (5.2%) HBsAg seroconversion at the end of 12 months and there were four (21.05%) patients who responded positively to vaccine therapy in this group. Asymptomatic carriers and controls did not have seroconversion of HBs Ag. Also, HBV DNA did not become negative in controls. CONCLUSION: It may be concluded that recombinant hepatitis B vaccine is effective in the treatment of chronic hepatitis B.     

慢性乙型肝炎病毒感染者前S1抗原和抗体检测的临床意义

目的 探讨前S1抗原(PreS1Ag)与前S1抗体（抗-PreS1）在慢性HBV感染者中的检出情况及其临床意义。方法 收集慢性HBV感染者428例,检测HBV血清学标志物、HBV DNA及PreS1Ag、抗-PreS1,分析PreS1Ag、抗-PreS1在慢性HBV感染不同转归人群[即e抗原阳性慢性乙型肝炎(CHB)组、e抗原阴性CHB组、非活动性HBsAg携带组、HBsAg血清学转换组]的检出情况及临床意义;同时分析PreS1Ag、抗-PreS1与HBV标志物、HBV DNA的关系。用SPSS13.0软件进行统计学处理。计数资料采用四格表或配对x2检验,计量资料采用独立样本t检验或秩和检验进行数据分析。结果 PreS1Ag阳性率e抗原阳性CHB组为95.7％(67/70)、e抗原阴性CHB组为82.8％ (24/29)、非活动性HBsAg携带组为13.2％(7/53)、HBsAg转换组为2.2％ (1/46),四组人群PreS1Ag阳性率依次下降,x2=141.7,P＜0.05,总体比较差异有统计学意义,可见PreS1Ag阳性率随病毒复制活跃程度下降而降低。抗-PreS1在四组间检出率差异无统计学意义。将HBsAg阳性的前三组（即e抗原阳性CHB组、e抗原阴性CHB组、非活动性HBsAg携带组）合并为一组,再与HBsAg血清学转换组比较,抗-PreS1阳性率分别为0.9％(14/152)和23.9％0 (14/46),x2 =6.919,P＜0.05,差异有统计学意义。PreSl Ag的吸光度值的平均秩次在高复制组为179.30,低复制组为133.80,高复制组明显高于低复制组,Z=-3.86,P＜0.05,差异有统计学意义;虽两组抗-PreS1的吸光度值平均秩次差异无统计学意义,但低复制组(23.86)较高复制组(21.08)有升高趋势。通过配对计数X2检验分析抗-HBs与抗-PreS1的吻合性,x2=0.262,P＞0.05,两者检出率差异无统计学意义。血清PreS1Ag、HBeAg、肝组织内HBcAg与HBV DNA有关联性,x2值分别为33.840、24.159、4.854,P值均＜0.05。血清PreS1Ag与HBV DNA的关联程度(r=0.628)高于HBeAg(r=0.563)。 结论PreS1Ag较HBeAg更敏感的反映了HBV复制的情况,抗-PreS1可能参与了HBV的清除,预示着慢性乙型肝炎的恢复。     

In situ detection of mutated hepatitis B virus in microdissected, formalin-fixed liver tissues from patients with chronic hepatitis B

BACKGROUND/AIMS: Hepatitis B virus (HBV) quasispecies have been detected in patients with chronic hepatitis B. In order to elucidate the relationship between HBV mutation and liver cell necrosis in situ, we analyzed sublobule-sized specimens microdissected from formalin-fixed paraffin-embedded liver biopsy tissues taken from patients with chronic hepatitis B. METHODS: The subjects were 20 patients with chronic hepatitis B. We extracted HBV-DNA from two sublobular regions of HBV-infected liver biopsy tissue, those with the most severe and the mildest hepatitis activity, demonstrated microscopically. The DNA coding sequence of the precore-core region of HBV was determined by amplifying the DNA by the polymerase chain reaction, followed by direct sequencing. RESULTS: In all seven patients with minimal to mild hepatitis activity, but only 4 of 13 with moderate to severe activity, the amino acid sequence of the precore-core region of HBV obtained from the region with the most severe hepatitis activity showed over 99% homology with the corresponding sequence of HBV obtained from region with the mildest hepatitis activity (p<0.05). CONCLUSION: The differences between intrahepatic HBVs observed in patients with highly active hepatitis suggest that exacerbation of hepatitis in vivo is related to the appearance of variants in the precore-core region of HBV.     

Changes in serum albumin during treatment of chronic hepatitis B with lamivudine: effects of response and emergence of drug resistance

OBJECTIVES: In chronic hepatitis B patients treated with lamivudine, the incidence of drug resistance increases with the duration of therapy. The effect of drug resistance on hepatic synthetic function is not well defined. The aim of the present study was to assess the effect of lamivudine therapy on hepatic synthetic function in patients with moderately severe chronic hepatitis B and, particularly, to determine the effect of drug resistance. METHODS: Hepatic synthetic function was assessed using serial measurements of serum albumin in 38 patients (26 with cirrhosis) in an open-label treatment program. RESULTS: An initial antiviral response (hepatitis B virus [HBV] DNA undetectable by hybridization assay) occurred in all patients, and nine of 22 (41%) hepatitis B e antigen-positive cases underwent hepatitis B e antigen seroconversion. Among 29 patients with undetectable serum HBV DNA at the end of observation, the mean serum albumin concentration rose from 39.9 +/- 0.7 to 43.2 +/- 0.6 g/L, corresponding to a yearly increase of 1.85 g/L (p < 0.001). This was largely attributable to an increase among cirrhotic patients. Nine patients (24%) developed resistance to lamivudine, all after 12 months of treatment. Among them, the mean serum albumin concentration had increased from 39.6 +/- 1.2 to 42.9 +/- 0.8 g/L before resistance emerged, but then decreased to 39.3 +/- 1.7 g/L (p = 0.01) at the time of reappearance of HBV DNA. CONCLUSION: Suppression of viral replication by lamivudine improves hepatic synthetic function in chronic hepatitis B patients, but emergence of drug resistance is associated with a rapid decline in serum albumin, at least to pretreatment levels.