Difference of subtypes of hepatitis B surface antigen between asymptomatic carriers and patients with chronic liver diseases

Subtypes of hepatitis B surface antigen (HBsAg) were studied in 20 asymptomatic HBsAg carriers and 18 HBsAg-seropositive patients with chronic hepatitis or live cirrhosis. Among them 25 were university students. In the male university students, subtypes adw was found to be significantly higher in asymptomatic HBsAg carriers than in patients with chronic hepatitis (p=0.04). The difference in subtypes of HBsAg between male asymptomatic carriers and male patients with liver disease was further confirmed by the combined results of university students and other subjects (p=0.007).     

Chronic hepatitis B and hepatitis B surface antigen carriers in university students

A study was made of latent hepatic diseases in university students. 1.4% to 1.6% of the students were positive for hepatitis B surface antigen (HBsAg), and 10.3% for anti-HBs. Of 28 students with HBs-antigenemia, 2 had chronic persistent hepatitis, and 3 minimal hepatitis, 23 being healthy carriers. Hepatitis B e-antigen (HBeAg) was detected in 44% of the students with HBsAg, and anti-HBe in 13%. Anti-HBe was significantly more frequently found in female students with HBsAg than in male students. Though most of the students with HBsAg had high titer of antibody to hepatitis B core antigen (anti-HBc), there were a small number of cases showing low titer. HBsAg and anti-HBs was detected in the same serum specimens of 2 carrier students. Liver damage was also found in 3 students without HBs-antigenemia.     

The role of IgM anti-HBc in the etiologic diagnosis of HBsAg-positive acute and chronic hepatitis

Summary HBsAg, HBeAg, anti-HBe, anti-HBc, anti-HBs, IgM anti-HAV, total and IgM anti-delta (RIA) and IgM anti-HBc (ELISA) have been determined on sera of 72 male patients with acute hepatitis 68 HBsAg-positive and 4 anti-HBc-positive) at the onset of the disease and at 2, 4 and 6 months thereafter, and of 77 patients with HBsAg-positive chronic active hepatitis. The results indicate that testing for IgM anti-HBc in acute hepatitis is necessary to identify cases of hepatitis B even in the absence of positivity for HBsAg and/or HBeAg. HBsAg-positive cases, negative for IgM anti-HBc, were due to delta superinfection of chronic carriers of HBsAg. Recovery was always associated with negativity of IgM anti-HBc; IgM anti-HBc persistence was not found in all cases evolved to chronicity. In chronic hepatitis cases positivity for IgM anti-HBc was found only in 39% of the anti-delta-negative and only in 5.5% of the anti-delta-positive cases. This work was supported in part by grant no. 82.02235.04 from theConsiglio Nazionale delle Ricerche (CNR), Roma, Italy.     

不同类型乙型肝炎患者HBV基因型的分布

目的 探讨乙型肝炎病毒（HBV）各种基因型在不同类型乙型肝炎患者中的分布.方法 用PCR结合限制性片段长度多态性（PCR-RFLP）方法检测56例HBV感染的肝癌、58例肝硬化、60例慢性乙型肝炎患者以及60例HBV无症状携带者血清HBV基因型.结果 无症状携带者中B基因型为46.7%（28/60）,C基因型为33.3%（20/60）,D基因型为20.0%（12/60）;慢性乙型肝炎组B基因型为41.7%（25/60）,C基因型为36.7%（22/60）,D基因型为21.7%（13/60）;肝癌患者中B基因型为33.9%（19/56）、C基因型为60.7%（34/56）、D基因型为5.4%（3/56）;肝硬化患者中B基因型为31.0%（18/58）、C基因型为63.8%（37/58）,D基因型为3.5%（2/58）.结论 在慢性乙型肝炎和无症状携带者中以B基因型为优势感染毒株,而肝硬化和肝癌患者则以C基因型为优势感染毒株.     

Variations in codons 84-101 in the core nucleotide sequence correlate with hepatocellular injury in chronic hepatitis B virus infection.

Individuals with chronic hepatitis B virus (HBV) infection are generally divided into asymptomatic healthy carriers and patients with chronic liver disease. Several studies have suggested that the hepatitis B core antigen could be an immunological target of cytotoxic T lymphocytes (CTL). To investigate the possible pressure site from CTL, the entire core region of HBV DNA was sequenced in 30 subjects (10 asymptomatic healthy carriers and 20 patients with chronic liver disease). No significant changes in the nucleotide sequence and deduced amino acid residue were noted in the 10 healthy carriers. In contrast, a cluster of changes in a small segment of 18 amino acids (codons 84-101 from the start of the core gene) was found in 15 of the 20 chronic liver disease patients. All these 15 patients had advanced liver diseases (chronic active hepatitis and cirrhosis), whereas only mild liver disease (chronic persistent hepatitis) was found in the five patients without mutations. These data suggest that the region with mutation clustering is the major target of CTL, and that the mutations evolve under the pressure of immune selection.     

Synthesis of antibodies to hepatitis B virus by cultured lymphocytes from chronic hepatitis B surface antigen carriers

It has been postulated that host immune defects are responsible for the development and persistence of the hepatitis B surface antigen (HBsAg) carrier state. The nature of these defects is unknown, but the absence of a readily detectable antibody response to HBsAg (anti-HBs) may be important. The synthesis of both anti-HBs and antibody to hepatitis B core antigen (anti-HBc) in cultures containing peripheral blood mononuclear cells from chronic HBsAg carriers and from control (antibody-positive) patients was measured in the presence of pokeweed mitogen. Similar amounts of polyclonal IgG and IgM were synthesized by cultures containing lymphocytes from chronic carriers and controls. Anti-HBc was detectable in lymphocyte supernatants from 2 of 20 controls and from 21 of 29 carriers. The presence of anti-HBc synthesis in vitro correlated with high serum titers of anti-HBc. In contrast, anti-HBs was detected in lymphocyte supernatants from 6 of 20 controls (predominantly in those who had high serum titers of anti-HBs) but in none of the supernatants from 29 HBsAg carriers. In order to identify the mechanisms for the lack of detectable anti-HBs synthesis by chronic HBsAg carrier lymphocytes, co-culture experiments were performed using T and B lymphocyte fractions that had been purified by affinity chromatography. B lymphocytes from carriers co-cultured with allogeneic irradiated ("helper") T lymphocytes from controls synthesized normal amounts of IgG, IgM, and anti-HBc but still did not synthesize detectable amounts of anti-HBs. In the converse experiments, B lymphocytes from controls were co-cultured with irradiated T lymphocytes from carriers. The T lymphocytes from 16 of 24 carriers augmented anti-HBs production by control B cells normally, the remaining eight did not. Finally, mixtures of control B cells and control irradiated T lymphocytes were co-cultured with T lymphocytes from chronic HBsAg carriers. 5 of 12 carriers demonstrated active suppression of anti-HBs production, and in three this suppression was specific, as IgG and IgM production remained normal. We conclude that chronic HBsAg carriers have a specific B lymphocyte defect in anti-HBs production. In addition, defects in the function of regulatory T lymphocytes may contribute to the absence of anti-HBs synthesis in some HBsAg carriers.     

Expression of HBsAg on the membrane of hepatocytes in chronic carriers of hepatitis B virus infection.

Liver cell membrane localization of hepatitis B surface antigen (HBsAg) was investigated in 31 asymptomatic chronic carriers by a direct immunofluorescence technique. A close relationship was found between absence of inflammatory liver disease, presence of large amounts of HBsAg in the liver and expression of the antigen at the hepatocyte surface. Capping of HBsAg after the addition of anti-HBs serum could be inhibited by factors (temperature, metabolic inhibition) that are known to influence viral antigenic mobility at the cell surface. In two patients with chronic active hepatitis as well as in some cases showing histological features of focal parenchymal necrosis, HBsAg could be detected in the cytoplasm of a few scattered hepatocytes but never at the surface of the cells. Both the cases with CAH and one with focal parenchymal necrosis had IgG bound to the liver cell membrane. These findings are in agreement with the hypothesis that the absence of liver damage in HBsAg healthy chronic carriers is related to immune tolerance to the antigen. In chronic active liver disease the presence of IgG on the membrane of hepatocytes suggests a possible role of blocking antibodies directed against viral antigens expressed at the hepatocyte surface.     

Hepatitis B core antibody screening of voluntary blood donors: an extended pilot study using a modified passive haemagglutination assay

Summary. Screening for hepatitis B surface antigen (HBsAg), even by the most sensitive techniques, fails to detect all carriers of the hepatitis B virus (HBV). The presence of hepatitis B core antibody (anti-HBc) in the absence of HBsAg is a common finding in donors implicated in cases of post-transfusion hepatitis B (PTHB), and viral DNA may also be demonstrated in many of these individuals. An extended pilot study of routine screening of all donations for anti-HBc in addition to HBsAg was introduced into the Mersey and North Wales Regional Transfusion Service in November 1991 to improve surveillance for carriers of HBV. In order to reduce costs a modified passive haemagglutination assay was evaluated and found to have a sensitivity of 99% and specificity of 98% compared with a conventional assay. In the first 6 months 60 530 donations were tested and 12 (0–02%) were found to have anti-HBc in the absence of HBsAg or adequate antibodies to HBsAg (anti-HBs). These sera will later be submitted for polymerase chain reaction (PCR) testing in order to determine their infectivity or otherwise by the detection HBV DNA sequences.     

Seropositivity of hepatitis B e antigen and hepatocellular carcinoma

Chronic infection with hepatitis B virus (HBV) is an important clinical problem due to its worldwide distribution and potential of adverse sequelae including hepatocellular carcinoma (HCC). Hepatitis B e antigen (HBeAg) is a biomarker of active viral proliferation in hepatocytes and infectivity. The prevalence of HBeAg among subjects chronically infected with HBV decreases with the increase in age. Case series studies have found a lowest seroprevalence of HBeAg in HCC patients compared with patients affected with chronic hepatitis B and liver cirrhosis. Case-control studies have shown a significantly higher seroprevalence of HBeAg in HCC cases than matched controls. A recent long-term follow-up study has shown a significantly elevated HCC risk for seropositives of both hepatitis B surface antigen (HBsAg) and HBeAg compared with seropositives of HBsAg only and seronegatives. The biological gradient remained in further stratification analyses by serum level of alanine transaminase and status of liver cirrhosis detected by ultrasonography. The cumulative HCC risk from age 30 to 70 years has been estimated to be 87% for those who were persistently seropositive on HBsAg and HBeAg, 12% for those with persistent seropositivity of HBsAg only, and 1% for those who were seronegative on HBsAg and HBeAg.     

Seroprevalence of hepatitis B and C viruses in patients with chronic kidney disease in the predialysis stage at a university hospital in Turkey

BACKGROUND: Hepatitis B (HBV) and C (HCV) viruses are the most common viruses that cause viral infections among the hemodialysis patients. OBJECTIVES: To assess the prevalence of HBV and HCV in predialytic chronic kidney disease (CKD) patients. DESIGN: A cross-sectional study. SUBJECTS: 171 consecutive predialytic CKD patients. MEASUREMENTS: Third-generation micro-ELISA assay was used for hepatitis B surface antigen (HBsAg), antibody to hepatitis B core (anti-HBc) and surface antibody (anti-HBs), secretory form of hepatitis B envelop antigen (HBeAg), antibody to secretory form of hepatitis B envelop antigen (anti-HBe), and ELISA for antibody to hepatitis C virus (anti-HCV). RESULTS: The main causes of CKD were 29.8% diabetic nephropathy, 19.9% chronic glomerulonephritis, 16.3% hypertensive nephrosclerosis, 14.0% unknown, 5.3% amyloidosis, 4.7% autosomal-dominant polycystic kidney disease, 4.1% chronic tubuluointerstitial nephritis, 3.5% malignancies, 1.7% benign prostatic hypertrophy, 0.6% Alport syndrome. The seroprevalence of hepatitis was: HBsAg 10.5%, anti-HBc 36.8%, anti-HBs 28.7%, HBeAg 5.3%, anti-HBe 32.7%, anti-HCV 7% and HBsAg+anti-HCV 0.6%. CONCLUSIONS: The seroprevalence of HBsAg and anti-HCV among predialytic CKD patients was similar to our patients in hemodialysis program.     

Circulating immune complexes in HBsAg-positive and HBsAg-negative chronic liver diseases

Summary Circulating immune complexes (CIC) were investigated by the fluid-phase Clq binding test in serum samples from 131 subjects with various clinical types of HBsAg-positive and HBsAg-negative chronic hepatitis diagnosed on both clinical and laboratory criteria, including liver biopsy; serum samples from 28 asymptomatic HBsAg carriers and 75 healthy controls were also examined. The results of these findings were correlated with the liver damage and the clinical course of the disease. CIC were detected in all categories of chronic hepatitis with a significant prevalence only in patients with liver cirrhosis. No correlation was found between the leveis of CIC, the presence of circulating HBsAg, and active inflammation or necrosis in any of the different types of chronic hepatitis. In contrast, a highly significant correlation was found between the prevalence of CIC and a more severe prognosis in patients with CAH and liver cirrhosis. The findings suggest that the presence of CIC is not specific for a given type of chronic liver disease and that they do not play any role in the pathogenesis of liver damage. The correlation observed between the presence of CIC and an unfavourable course of chronic liver disease suggests that CIC may modify the host’s immune defence mechanisms. This work was supported in part by grants from theConsiglio Nazionale delle Ricerche (CNR) and from theMinistero della Pubblica Istruzione, Roma, Italy.     

Virological classification of autopsy livers with hepatic disorders

Forty six selected autopsy livers with hepatic disorders were classified histologically into three groups, i.e., hepatitis 8, liver cirrhosis 16 and hepatocellular carcinoma 22, chiefly by histological findings. These groups were subdivided into two categories after determining the presence or absence of three HBV associated markers, i.e., the hepatitis B surface antigen (HBsAg), the antibody to hepatitis B surface antigen (anti-HBs) and the antibody to hepatitis B core antigen (anti-HBc) in the sera and liver homogenates. At least one of these markers was found to be present in the sera and homogenates of 38% of the livers with hepatitis, 69% of the livers with liver cirrhosis and 77% of the livers with hepatocellular carcinoma. When the liver tissues were examined for the presence of HBV DNA using Southern blot technique, the majority of them (10 out of 11) which proved to be positive for at least one HBV associated marker were also positive for HBV DNA. However, HBV DNA could not be detected in the other 10 livers which contained no HBV associated markers. These results showed that a HBV associated serological marker was always expressed, when liver tissue HBV DNA was demonstrable. The results of the two detection methods we used in this study were found to be almost equivalent. These results showed no evidence of nucleic acid homology between DNA from the liver of patients with non-B type liver disorders and HBV DNA.     

各类慢性乙型肝炎患者HBV前S2抗原与HBV感染血清标志物及HBVDNA的关系

目的探讨乙型肝炎病毒(HBV)前S2抗原(preS2Ag)与HBV感染5项标志物(HBV-M)、HBV DNA之间的关系及对慢性乙型肝炎分类诊断的临床意义。方法对584例各类慢性乙型肝炎患者血清采用放射免疫法(R IA)检测preS2Ag,用酶联免疫吸附试验(ELISA)检测HBV-M(HBsAg、抗HBs、HBeAg、抗HBe、抗HBc),用聚合酶链反应(PCR)检测HBV DNA。结果慢性乙型肝炎轻度、中度、重度及肝炎后肝硬化患者preS2Ag的检出率分别为45.98%、67.10%、83.87%及92.86%;preS2Ag在HBeAg阳性、HBV DNA>105拷贝/mL患者中检出率明显高于HBeAg阴性、HBV DNA<105拷贝/mL患者,差异有统计学意义(P<0.001)。结论preS2Ag的检出意味着病毒有复制或有传染性;开展preS2Ag的检测有助于慢性乙型肝炎的分类、慢性乙型肝炎急性发作和预后的判断。     

Serum HBV-DNA (hepatitis B virus DNA) in acute and chronic hepatitis B infection

HBV DNA was measured in the sera of 69 patients with hepatitis B virus infections. Sixteen patients had acute hepatitis B, 24 had chronic active hepatitis (CAH), 6 had chronic persistent hepatitis (CPH), 5 had cirrhosis without CAH and 18 were asymptomatic HBsAg carriers. In patients with acute hepatitis B who recovered, HBV DNA was present in the serum transiently early in the illness. HBV DNA persisted in the serum in the two patients who developed chronic hepatitis. Sera of 23 of 24 patients with CAH were persistently positive for HBV DNA. There was no relationship between the quantity of HBV DNA in the serum and the histological intensity of activity. Thirteen of the 24 patients with CAH had histological evidence of cirrhosis in addition to CAH and HBV DNA was detected in the sera of all 13. The sera of 2 of 6 patients with CPH were positive for HBV DNA. In one it was positive only where there was clinical evidence of reactivation of HBV infection. The other patient subsequently developed CAH. Sera of 5 patients with established HBsAg positive cirrhosis but without evidence of CAH were negative for HBV DNA. Two of these patients had hepatocellular carcinoma. Sera of 18 asymptomatic anti-HBe positive carriers with normal ALT were negative for HBV DNA. HBeAg and HBV DNA were not always found in the serum together. In acute hepatitis 5 patients with HBV DNA in the serum were HBeAg positive, but in 6 patients the sera were HBeAg positive inthe absenceof HBV DNA.     

Sequential accumulation of the basal core promoter and the precore mutations in the progression of hepatitis B virus-related chronic liver disease

OBJECTIVES: Despite the pathogenic role of the basal core promoter (BCP) and the precore mutations in chronic hepatitis B virus (HBV) infection, their role in the progression of liver disease is still controversial. We analyzed whether the accumulation of these mutations might enhance the progression of HBV-related chronic liver disease. METHODS: Forty consecutive patients at each clinical status were analyzed. Clinical statuses were as follows: HBeAg-positive asymptomatic carrier (HBeAg(+) ASC) (defined as HBeAg(+), anti-HBe(-), HBV-DNA(+) by hybridization, normal ALT); inactive HBsAg carrier; chronic hepatitis B; liver cirrhosis. The genotype and the BCP/precore regions were determined by PCR using genotype specific primers and direct sequencing, respectively. RESULTS: All patients except one were infected with genotype C. The A to T mutation at nucleotide 1762 and/or G to A mutation at nucleotide 1764 were found in 30% in HBeAg(+) ASC, 65.7% in inactive HBsAg carrier, 95% in chronic hepatitis B, and 90% in liver cirrhosis (p < 0.001). The prevalence of the G to A mutation at nucleotide 1896 was 5% in HBeAg(+) ASC, 22.5% in inactive HBsAg carrier, 32.5% in chronic hepatitis B, and 50% in liver cirrhosis, respectively (p < 0.001). The T to C/A mutation at nucleotide 1753 in the BCP and G to A mutation at nucleotide 1899 in the precore were more frequent in liver cirrhosis than in the other clinical statuses (p < 0.05). CONCLUSION: Sequential accumulation of mutations in the BCP/precore has an important role in the progression of HBV-related liver disease.     

Seropositivity of hepatitis B e antigen and hepatocellular carcinoma

Chronic infection with hepatitis B virus (HBV) is an important clinical problem due to its worldwide distribution and potential of adverse sequelae including hepatocellular carcinoma (HCC). Hepatitis B e antigen (HBeAg) is a biomarker of active viral proliferation in hepatocytes and infectivity. The prevalence of HBeAg among subjects chronically infected with HBV decreases with the increase in age. Case series studies have found a lowest seroprevalence of HBeAg in HCC patients compared with patients affected with chronic hepatitis B and liver cirrhosis. Case‐control studies have shown a significantly higher seroprevalence of HBeAg in HCC cases than matched controls. A recent long‐term follow‐up study has shown a significantly elevated HCC risk for seropositives of both hepatitis B surface antigen (HBsAg) and HBeAg compared with seropositives of HBsAg only and seronegatives. The biological gradient remained in further stratification analyses by serum level of alanine transaminase and status of liver cirrhosis detected by ultrasonography. The cumulative HCC risk from age 30 to 70 years has been estimated to be 87% for those who were persistently seropositive on HBsAg and HBeAg, 12% for those with persistent seropositivity of HBsAg only, and 1% for those who were seronegative on HBsAg and HBeAg.     

Familial clustering and immune response in family contacts of patients with HBsAg-positive liver cirrhosis

Families of 11 patients with hepatitis B surface antigen (HBsAg)-positive cirrhosis were studied to evaluate the immunologic correlates and extent of intrafamilial HBsAg clustering. Of 76 family contacts, 12 were identified to be asymptomatic carriers of HBsAg and two were diagnosed to have HSsAg-positive cirrhosis. The over-all HBsAg prevalence for the 76 contacts was 18% and that for all 87 members studied 29.0%. Serologic evidence of hepatitis B virus (HBV) infection (either HBsAg, anti-HBs, or both) was detected in 59% of all family members. HBsAg was more prevalent in males (47%) compared with females (16%), and anti-HBs was more prevalent in females (42%) compared with males (18%). Evidence of an immunologic response in clinically unaffected HBsAg-negative family contacts consisted of elevated serum IgG levels (mean 1660 mg/100 ml) and increased prevalence of anti-smooth muscle and antimitochondrial antibodies (16% and 6%, respectively). The prevalence of one or more autoantibodies in all HBsAg-negative family contacts was 20%, and it was higher in females (25%) than in males (13%). The present study demonstrates that HBsAg clustering occurs in families of patients with cirrhosis in the Jerusalem area and indicates that HBsAg-negative family contacts may have increased B-cell activity.     

The immunological diagnosis of HBsAg liver disease by combined screening for thee system in the serum and the HB core and surface antigens in liver biopsy samples

Summary Serological screening for HBeAg and anti-HBe, combined with the immunohistochemical localization of the hepatitis B core and surface antigen in 110 HBsAg carriers, established different immunological profiles indicative of the liver status and prognostic of the future absence or progression of the disease. Intrahepatic HBcAg and serum HBeAg appeared the most sensitive indexes of chronic and progressive liver disease, anti-HBe and large amounts of cytoplasmic HBsAg suggest, instead, the asymptomatic carrier state without liver damage. Only the nuclear localization of HBcAg in immunohistochemical studies was a reliable prognostic indicator of transition to chronicity; absence or presence of serum HBeAg was of no help in predicting the outcome of acute HBsAg hepatitis.     

Serological markers of hepatitis A and B virus infection in university students

Serological markers of hepatitis A and B virus infection were determined by radioimmunoassay in 162 university students. Antibody to hepatitis B e antigen (anti-HBe) was demonstrated in 11 of 15 students positive for both antibody to hepatitis B e antigen (HBsAg) and antibody to hepatitis B core antigen (anti-HBc). The presence of anti-HBc alone seemed to denote previous hepatitis B virus infection. Antibody to hepatitis A virus was demonstrated only in 3%. Study of 23 stored HBsAg-positive sera showed high titer anti-HBc significantly more frequently in sera positive for anti-HBe than in sera positive for hepatitis B e antigen. Incidence of hepatitis B e antigen was significantly higher in male students with HBsAg, and that of anti-HBe in female students.     

Hepatitis B core antigen antibody as an indicator of a low grade carrier state for hepatitis B virus in a Saudi Arabian blood donor population

Blood donor screening for anti-hepatitis B core antigen (anti-HBc) was introduced as a surrogate marker of non-A, non-B hepatitis prior to the availability of a specific test for hepatitis C. In areas endemic for hepatitis B virus (HBV), such as Saudi Arabia, earlier studies indicated that up to 30% of blood donors might disqualify if screened for anti-HBc. The issue was readdressed in a study of 6035 consecutive first-time Saudi national blood donors in an attempt to identify a subgroup of anti-HBc positive donors who might be at high risk of being low grade carriers of HBV. An isolated anti-HBc of high titer in a donor with a low or absent anti-hepatitis B surface antigen (anti-HBsAg) was taken as an indicator of increased risk of a low grade carrier state. Using this algorithm, an additional 125 (2%) donors would disqualify. HBsAg immune complex assays and polymerase chain reaction of donor samples with an isolated anti-HBc identified two donors with immune complexes and two donors with HBV DNA. All four donor samples expressed over 90% neutralization in the anti-HBc supplementary testing, indicating high titer anti-HBc. These findings seem to support the suggested policy of donor exclusion based on the anti-HBc and anti-HBsAg serology as a means to eliminate low grade carriers of HBV in endemic areas without jeopardizing the blood supply.