Enhancement of HBsAg detection in serum of patients with chronic liver disease following removal of circulating immune complexes

Patients with chronic liver disease and hepatocellular carcinoma may lack serological evidence of previous hepatitis B virus infection. The purpose of the present study was to test the hypothesis that circulating immune complexes may interfere with the detection of low levels of HBsAg in such patients. Sera from 190 patients were initially screened for the presence of circulating immune complexes. Patients belonged to three clinical categories: asymptomatic HBsAg carriers (50 patients), chronic liver disease (30 patients) and hepatocellular carcinoma (110 patients). Forty-one of the group of 190 patients (21%) were positive for circulating immune complexes. Sera from 21 patients were selected for further evaluation. The sera of 13 chronic liver disease or hepatocellular carcinoma patients (HBsAg negative, hepatitis B virus-DNA negative, with or without evidence of previous hepatitis B virus infection) and eight HBsAg positive carriers (four asymptomatic, three with chronic liver disease and one with hepatocellular carcinoma) were passed through a Clq affinity column (first column) to remove circulating immune complexes. Unbound material was then passed through a monoclonal IgG2a anti-HBs affinity column (second column). Unbound material (following both columns) contained free HBsAg, as determined by monocolonal radio-immunoassay, in eight patients in whom HBsAg had been undetectable in the original serum. Removal of circulating immune complexes from the serum of the three HBsAg positive patients with chronic liver disease also caused a significant increase in measurable circulating HBsAg compared with the original serum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immune response to HBsAg, HBcAg and e-antigen in patients with acute hepatitis and HBsAg carriers with and without liver diseases

Humoral and/or cell-mediated (CMI) immune responses to HBAg components, human and rabbit liver specific proteins (HLP and RLP) and tuberculin were tested in patients with acute virus B and non-B-hepatitis, asymptomatic HBsAg carriers and HBsAg positive chronic active hepatitis (CAH). Furthermore, the presence of HBsAg, HBcAg and/or "e"-antigen has been studied in patients with sera and/or liver tissue. Asymptomatic HBsAg carriers are characterized by a status of immunological tolerance against HBsAg. HBcAg in liver nuclei could not be detected. All sera were positive for anti-HBc, some had anti "e". - Patients with uneventful acute virus-B-hepatitis developed CMI against HBsAg 4-6 weeks and anti-HBs 4-6 months after onset of the disease. Acute virus hepatitis without detectable HBsAg are defined as non-B-hepatitis by negative humoral and cell-mediated immune reaction against HBsAg 1-12 months after onset of the disease. - Patients with type B chronic active hepatitis are characterized by inadequate CMI against HBsAg without immune elimination of virus and virusantigens. Acute and chronic type-B-hepatitis showed temporary or constant CMI against HLP. These findings suggest an alteration or a carrier function of membrane antigens of virus infected hepatocytes or an induction of new membrane antigens by a virus. The results indicate that recovery from type B-hepatitis is associated with the ability to elicit a specific immune response to HBsAg. Furthermore immune responses to virus, virus antigens and virusinfected hepatocytes seemed to be the pathogenic principle of virus induced acute and chronic liver diseases.     

The effect of interleukin-1alpha on outflow facility in rat eyes

BACKGROUND: There are few reports in Mexico on the prevalence of infection by virus D. OBJECTIVE: The aim of the present study was to study the hepatitis D virus infection prevalence in patients entering to the University Hospital. METHODS: Seventy three HBsAg positive patients sera were studied. There were 38 patients with acute hepatitis, 28 patients with chronic liver disease and 7 were asymptomatic HBsAg carriers. Serological markers for hepatitis viruses B, D and C were detected by means of ELISA test (Abbott). RESULTS: Anti-HDV was detected in 3 cases (4%). The first two cases were men with acute hepatitis B. Both had a coinfection by viruses B and D, however IgM anticore could not be demonstrated in the first case, this patient developed hepatic cirrhosis within 13 months, in addition he had a concurrent infection by hepatitis C virus with a positive second generation ELISA antibody. The second case recovered from the acute hepatitis. The third case was a female nurse with acute hepatitis and a coinfection by viruses B and D who recovered from the acute attack. Antibody to hepatitis C was present in 3 out of 22 patients with chronic liver disease (13.6%), one of them having an hepatocellular carcinoma. CONCLUSIONS: Our results coincide with the previously reported low incidence of hepatitis D and represent the first report in Mexico of concurrent infections by viruses B-C and B-D-C.     

Effect of training on general practitioners'' use of a brief intervention for excessive drinkers

During recent years the outcome of acute hepatitis A in chronic liver disease has been discussed controversially. Data from large hepatitis A epidemics and surveillance data from the United States suggest a significantly higher risk of fatal outcome in patients with chronic hepatitis B. Patients with chronic active hepatitis or liver cirrhosis seem to be at highest risk, while HBsAg carriers may exhibit a benign course of the disease. Patients with chronic hepatitis C also seem to have a significantly higher risk of fulminant hepatic failure when superinfected with hepatitis A. The recently reported unsuspected coincidence of autoimmune markers with a fulminant course of hepatitis A in those patients needs to be confirmed. Vaccination against hepatitis A in patients with chronic liver disease has been shown to be safe and effective.     

Identification of hepatitis B virus-DNA in the liver by in situ hybridization using a biotinylated probe. Relation to HBcAg expression and histology

The cellular localisation of hepatitis B virus (HBV)-DNA in liver tissue was studied by in situ hybridisation using biotinylated and radiolabelled probes on samples from HBsAg carriers with a spectrum of disease and related to the presence of HBV-DNA in serum and intrahepatic HBcAg expression. Sixteen of the 31 patients studied were seropositive for HBV-DNA; nine had chronic active hepatitis and seven had chronic persistent hepatitis. HBV-DNA was detected in the liver tissue in seven of these patients. In each, HBV-DNA was detected in both cytoplasm and nuclei. All seven also had nuclear and/or cytoplasmic HBcAg which in six was associated with chronic active hepatitis. HBcAg (without tissue HBV-DNA) was detected in the remaining nine patients with an exclusively nuclear pattern in two. Fifteen patients were seronegative for HBV-DNA. HBV-DNA was not detected in the tissue of any of these. Three of these were HBcAg positive but in each this was confined to occasional nuclei and each had inactive disease. The close association between the presence of detectable HBV-DNA in tissue, cytoplasmic HBV-DNA expression and chronic active hepatitis in one group and a failure to detect HBV-DNA in those with nuclear HBcAg and benign disease suggests that there may be two distinct patterns of HBV replication in chronic HBV carriers which may influence the development of liver damage.     

Nested polymerase chain reaction (PCR) and multiple cloned antibody capture PCR for the examination of serum hepatitis B virus DNA in negative hepatitis B surface antigen patients suffering from liver diseases

In order to find out rapidly the causes of the liver diseases suffered by patients with negative hepatitis B surface antigen (HBsAg), nested polymerase chain reaction (PCR) and multiple cloned antibody capture PCR techniques were established to examine serum hepatitis B virus (HBV) DNA. By using both techniques along with the examination of hepatitis C virus (HCV) infection, the causes of chronic liver diseases with negative HBsAg were studied. It is found that nested-PCR can increase the sensitivity of single PCR more than 1,000 fold and multiple cloned antibody capture-PCR can detect concentration of HBV DNA as low as 0.1-0.01 pg/L. HBV DNA positive patients were found in 45.5%, 30.8%, 13.3% and 100% respectively of the patients suffering from liver cirhosis with negative HBsAg (group A, 22 cases), chronic hepatitis with negative HBsAg (group B, 13 cases), normal subjects with negative HBsAg and positive hepatitis B core antibody (HBcAb, group C, 30 cases) and liver cirhosis with positive HBsAg and negative HBeAg (group D, 12 cases). HBV DNA can be also found in the serum of HBsAb positive patients and subjects supposed to be healthy, 81.8% and 53.8% of the patients were infected with HBV and/or HCV in group A and group B respectively. All these results suggest that nested-PCR and multiple cloned antibody capture-PCR are rapid and highly sensitive methods for detection of serum HBV DNA. HBV infection is an important cause of chronic liver diseases in patients with negative HBsAg. The causes of most of the HBsAg-negative chronic liver diseases are related with infection of viruses. The clinical significance of serum HBsAb in naturally infected patients should be reconsidered.     

Construction and characterisation of Salmonella typhimurium aroA simultaneously expressing the five pertussis toxin subunits

Viral hepatitis belongs to the most important infectious diseases worldwide. More than 300 million chronic HBsAg carriers and chronic HCV carriers exist, respectively. High endemic areas of viral hepatitis are Asia, Africa, Latin America and the Near, Middle and Far East. Viral hepatitis is also very important in health care workers. Today viral hepatitis can be differentiated from type A to type E (G) based on immunological and molecular assays. While enterally transmitted hepatitis type A and type E only induce acute and rare fulminant disease, hepatitis type B, C and D often induce chronic progressive disease including liver cirrhosis with typical complications due to the portal hypertension and with a high rate of association with the development of primary liver cancer (HCC). This review focusses on viral hepatitis-related surgical problems, including liver transplantation.     

Genital infections and their care

The pathogenesis of hepatitis B can be subdivided into three sequentially correlated events: (a) loss of virus tolerance, (b) liver cell necrosis mediated by virus specific inflammatory response, (c) non-specific death of functionally compromised hepatocytes mediated by inflammatory cytochines released by virus specific inflammatory response. The severity of liver damage depends on the occurrence of these events as well as other factors. The HBeAg defective mutant appears to be involved in the loss of virus tolerance and therefore in the pathogenesis of acute hepatitis B. In addition it is positively selected by antiviral immunoreaction, behaves as an escape mutant, and it also contributes to the pathogenesis of chronic hepatitis B. The combination of these characteristics explains the relative prevalence of this mutant over wild-type HBV in patients with severe acute hepatitis B and in chronic HBsAg carriers during anti-HBe seroconversion and/or hepatitis B exacerbations. However, the absence of HBeAg defective mutants in some cases of severe and fulminant hepatitis B as well as its detection in asymptomatic carriers of HBsAg should not be surprising. The severity of hepatitis is influenced by many other factors: the number of virus infected cells, the competence and genetic heterogeneity of the immune system, the vigor and extent of non-specific inflammatory response and the killing of hepatocytes endangered by other diseases or infected with other hepatotropic viruses.     

The intrahepatic expression of the hepatitis B virus in chronic delta hepatitis

In order to evaluate the interference of hepatitis delta virus (HDV) in hepatitis B viral particle (HBsAg, HBcAg) expression in the liver of chronic HDV patients, 39 and 81 liver biopsies of HBsAg carriers seropositive for anti-HDV and anti-HDV negative controls, respectively, were studied. HBcAg was positive in 16.7% of the HBeAg-positive patients with HDAg in the liver and in 91,4% of controls. In contrast, in HBeAg- and anti-HDV negative patients the intrahepatic expression of HBcAg was detected in 32.6%. In anti-HDV negative patients the HBcAg liver expression correlated significantly with the HBeAg in serum (p < 0.00001). The distribution of HBcAg was exclusively cytoplasmatic in 30% of HDV-infected patients but mixed nuclear and cytoplasmic in 38.3% of the controls. The nuclear expression of HBcAg was decreased in chronic HDV infection. HBsAg was positive in 70.3% of patients who were anti-HDV positive and in 82.3% of controls. The membranous expression of HBsAg was detected less frequently in HDV-infected patients (p < 0.05) than in controls, while associated with HBeAg in serum of HBV carriers without HDV superinfection (p < 0.00001). The prevalence and the HBsAg cytoplasmic expression was not different for the chronic HDV infection or controls. Our results show: 1) decreased intrahepatic expression of HBcAg and membranous HBsAg in HBV carriers superinfected with HDV, suggesting decreased HBV replication in the liver of these patients. 2) the changing of HBcAg and HBsAg expression in the liver of HDV-infected patients, suggest not so much a decrease but rather a modulation in HBV replication.     

Serial liver biopsy observations in hepatitis B antigen carriers by light and electron microscopy

Liver morphology and function were restudied after an interval of 1 1/2-2 years in 21 Chinese men with hepatitis B antigen (HBsAg). Antigenemia and mild liver histopathology disappeared in one individual who developed antibody to HBsAg(Anti-HBsAg). 20 subjects who were chronic carriers of HBsAg had mild histological abnormalities which were usually persistent and were more commonly lobular than portal. Particularly striking were focal necrosis, eosinophilic bodies, ground-glass cytoplasmic change, and, by electron microscopy, microtubular structures within cytoplasmic membranous cysternae. Interval development of chronic aggressive hepatitis was observed in only one subject. This study suggests that the great majority of HBsAg carriers have a good prognosis at least over a 1 1/2-2-year period, and that liver biopsy is most indicated in those with persistent liver function abnormalities.     

The effect of praziquantel administration on the course of hepatitis B among cases with concomitant schistosome infection

The present work was designed to study the course of HB virus infection among bilharzial and non-bilharzial patients and to assess the therapeutic effect of praziquantel administration on subsequent course of HB among individuals with concomitant infections. This study included 26 bilharzial cases, 14 cases with HB and 40 cases with both infections (HB and schistosomiasis). Praziquantel was administered to all bilharzial positive cases. Sera were collected from all groups prior to anti-bilharzial chemotherapy, and then later at three and six months post treatment. The results show improvement of both liver function tests and cell mediated immunity as estimated by increased mean value of CD3, CD4, helper/suppressor ratio among individuals who received praziquantel in the two groups with schistosomiasis and concomitant infection. Furthermore the individuals who lost their HB surface antigenaemia were found to have a higher mean pan T cells, CD4 values, normal helper/suppressor ratio and absence of Clc in their sera than those who retained their carrier state. The follow up of HB carriers demonstrated a higher cure rate (clearance of HBsAg) among the group with concomitant infection as compared to the group with virus hepatitis only. HBV type two infection was common among the study population accounting for 25.9% of HBsAg positive cases. 42.9% of them cleared their antigenemia after treatment with praziquantel.     

The effect of i.v. enalaprilat in chronically treated hypertensive patients during cardiac surgery

In areas where hepatitis B virus (HBV) is prevalent, HBV carriers negative for hepatitis B surface antigen (HbsAg) by enzyme-linked immunosorbent assay (ELISA) have been reported. Moreover, even after screening donor blood for HbsAg and hepatitis B core antibody (HBcAb), post-transfusion hepatitis B continues to occur, though with a decreasing frequency. Therefore, screening tests far more sensitive for detecting HBsAg than those currently available are needed. We developed a highly sensitive method for HBsAg detection. It is based on the recognition of peroxidase activity through measuring the formation of stable nitroxide radical with electron spin resonance (ESR) spectroscopy in the presence of hydrogen peroxide, p-acetamidophenol (p-AP), and 4-hydrazonomethyl-1-hydroxy-2,2,5,5,-tetramethyl-3-imidazoline-3-o xide (HHTIO). A cut-off value was established by testing of 186 healthy adults and 50 HBsAg-positive individuals. The signal to noise (S/N) ratio of less than 1.488 obtained by ESR spectroscopy was considered to be negative and more than 2.181, positive. The p-AP/HHTIO method was found to be 10 times more sensitive than the standard ELISA and reproducibility was excellent. Additional investigations were made on the HBsAg levels in the serum from 26 healthy subjects, in whom cut-off index levels on ELISA were negative but relatively high (range: 0.6 to 1.0); and on 15 patients with non B non C hepatitis. Three of 26 cases and 3 of 15 with non B non C hepatitis were judged to be HBsAg positive. Of these, 5 were found to be positive for HBV DNA by polymerase chain reaction (PCR). It was shown in this study that the p-AP/HHTIO method is practical and useful in screening HBV carriers because of the sensitivity in HBsAg detection, which is comparable to PCR analysis.     

The natural course of hepatitis B and hepatitis C virus infection

Chronic hepatitis B and hepatitis C virus infections maintain a significant risk for the development of liver cirrhosis and hepatocellular carcinoma and cause a considerable morbidity in the population. Among patients with chronic HBV infection and histologically confirmed hepatitis the annual incidence of liver cirrhosis is 2%. The risk for hepatocellular carcinoma in chronic HBsAg carriers is elevated about 40-230 fold. 20-30% of patients with chronic HCV infection will develop cirrhosis over 20-30 years. Hepatocellular carcinoma evolves yearly in about 3% of patients with chronic HCV infection and cirrhosis, whereas HCV-carriers without cirrhosis usually do not develop hepatocellular carcinoma. The high incidence of serious sequelae warrants a regular surveillance of chronic virus carriers.     

Factors associated with intrafamilial transmission of hepatitis B virus infection in Korea

Epidemiologic and serologic data on 137 household contacts of 51 chronic carriers of HBsAg and 111 household contacts of 38 controls who were negative for serologic markers of hepatitis B virus (HBV) were obtained from March 1990 to August 1991. Using this data, possible routes of intrafamilial transmission of hepatitis B virus among household contacts of chronic carriers of hepatitis B surface antigen (HBsAg) were evaluated and analyzed. The HBsAg prevalence among the household contacts of carriers was 14. 1% (95% CI 7.8-24.0) compared to 0.0% (95% CI 0.0-7.0) among those of controls (P < 0.01). The offspring of carriers showed significantly higher risk of HBV infection(relative risk; 6.6). Sharing of towels and handkerchieves, and drinking vessels was associated with an increased risk of HBV infection via intrafamilial transmission in Korea (relative risk 11.5 for towel and handkerchief, 12.1 for drinking vessels).     

The clinical olfactogram (author''s transl)

Sera from patients with acute hepatitis, cirrhosis or chronic hepatitis, as well as sera from healthy carriers and controls were examined for HBS antigen, DNA polymerase activity and for antibodies to HBS, HBC and DNA polymerase. The data presented suggest that in acute hepatitis the DNA polymerase test enabled us to diagnose at least 20% more cases of hepatitis B than with the RIA but that the DNA polymerase test is of little value for the screening of blood donors since all the healthy carriers gave negative results. As concerns the antibodies to DNA polymerase they appeared in at least 50% of the patients with acute hepatitis, they were transient and only detectable at the early beginning of the disease. These antibodies were also found to be different from the anti-HBS and anti-HBC antibodies.     

GB virus-C/hepatitis G virus infection in an area endemic for viral hepatitis, chronic liver disease, and liver cancer

BACKGROUND & AIMS: GB virus-C/hepatitis G virus (GBV-C/HGV) is a newly identified flavivirus, and little is known about its clinical significance. GBV-C/HGV was investigated in different populations, and its coinfection was investigated in patients with liver disease in Taiwan where hepatitis B and C are endemic. METHODS: Viral RNA was studied in 70 high-risk individuals, 20 patients with chronic non-B, non-C hepatitis, 13 with non-A-E fulminant hepatitis, 100 with asymptomatic hepatitis B surface antigen carriage, 120 with hepatitis B surface antigen-positive chronic liver disease and hepatocellular carcinoma, 100 patients with chronic hepatitis C, and 100 healthy adults. RESULTS: GBV-C/HGV infection was more frequent in high-risk groups (15%-30%) and hepatitis C virus carriers (10%) than in healthy adults (1%) and hepatitis B virus carriers (3.2%). Eighty-three percent of those infected had undergone blood transfusions previously. The prevalence in hepatitis B virus carriers increased with the severity of liver disease, being 1% in asymptomatic carriers and 10% in hepatocellular carcinoma. In chronic hepatitis C, clinical and virological data were comparable between those with and without coinfection. CONCLUSIONS: In Taiwan, GBV-C/HGV infection is common in high-risk groups, and its coinfection seems to not aggravate the course of chronic hepatitis B or C.     

Hepatitis-B virus and schistosomiasis infections in childhood proteinuria

Hepatitis-B surface antigen (HBsAg), circulating anti-schistosomal IgG (CSAb) and circulating specific schistosomal immune complexes (CIC) were detected, using ELISA, in sera of 40 active nephrotic children, 40 active S. mansoni infected cases and 20 apparently normal age-matched controls. The presence of HBsAg cases was significantly higher among nephrotic cases (20%), active S. mansoni cases (17.5%) than controls. Moreover, HBsAg cases were significantly higher in positive CIC S. mansoni cases than negative CIC ones. The mean O.D. readings of CSAb was significantly higher in positive HBsAg nephrotic cases than negatives. At the same time, the anti-schistosomal antibodies were higher in S. mansoni cases with proteinuria than those without. Specific CIC level was significantly higher among nephrotic and schistosomiasis cases than controls. The CIC were significantly higher in schistosomiasis cases with positive HBsAg than those with negative HBsAg and were detected in 80% of cases with proteinuria compared to 37% of cases without proteinuria with a statistically significant difference. On the other hand, CIC level was not influenced, in nephrotic cases, by the presence or absence of HBsAg. It was concluded that the presence of proteinuria was considered as a good monitor of the kidney affection either with schistosomiasis or the nephrotic syndrome or the HBsAg. The detection of CIC can be used as a good monitor too and could be included in methods of early diagnosis and/or following the disease prognosis.     

The relationship between hepatitis C virus and schistosomiasis: histopathologic evaluation of liver biopsy specimens

The reported high incidence of anti-HCV seropositivity in the Egyptian population seems surprising. Some suggest that schistosomiasis is the responsible factor, either by producing false positivity for HCV antibodies or by predisposing to actual HCV infection in some way. In an attempt to investigate this unclear relationship on a histological level, we performed a thorough semiquantitative morphological study of liver biopsy specimens from 44 anti-HCV-positive Egyptian patients with chronic liver disease. More than half of these patients (23) had serological evidence of schistosomiasis. The results have shown that all 44 liver biopsy specimens demonstrated the histopathological features known to be characteristic of chronic HCV hepatitis. Statistical analysis showed no significant difference between the schistosomal and nonschistosomal groups regarding the semiquantitative histological scores of these features. This study confirms the presence of definite HCV-induced hepatic pathology in all anti-HCV seropositive cases. More importantly, it shows the lack of enhancement of this pathology in the schistosomal patients.     

Seroepidemiological study of hepatitis delta virus infection in Okinawa, Japan

A seroepidemiological study on hepatitis delta virus (HDV) infection was conducted in the Okinawan islands, the area of Japan where hepatitis B virus infection is most prevalent. The subjects of this study included 116 asymptomatic hepatitis B surface antigen (HBsAg) carriers, 48 patients with chronic hepatitis (CH), 19 with liver cirrhosis (LC), and 11 with hepatocellular carcinoma (HCC). Among the 194 serum samples examined, a total of 10 (5.2%) were anti-HDV seropositive. Anti-HDV was detected in 2 (1.7%) of the 116 asymptomatic HBsAg carriers, in 3 (6.3%) of the 48 patients with CH, and in 5 (26.3%) of the 19 with LC. However, none of the patients with HCC had detectable anti-HDV. Eight of the 10 were born in the Miyako island group and the remaining 2 on the main island of Okinawa. Since the subjects included 34 individuals who were living and/or born in the Miyako islands, the positive rate of anti-HDV in the islands was 23.5%. This study demonstrates the existence of an endemic area of HDV infection in Japan.     

Berylliosis as an auto-immune disorder

Four random samples representing populations at low (volunteer blood donors), intermediate, (VD clinic patients), high (family contacts of chronic antigen carriers) and very high (male homosexuals) risk of exposure to HBV were surveyed. Among HBsAg and anti-HBs negative individuals an average of 3.3% were found to be anti-HBc positive, and among those with anti-HBs, 19.4% were anti-HBc positive. Anti-HBc, with concurrent anti-HBs and without, was detected more frequently in the high risk samples than in the low risk. Individuals was a past history of acute viral hepatitis were more frequently anti-HBc positive than those without such a history, and anti-HBc positivity was frequently accompanied by serum transaminase elevation. Anti-HBc may persist for many years after an episode of acute hepatitis. In households of carriers, the highest frequency of anti-HBc was observed among spouses, which would argue for the possibility of sexual transmission. A significant excess of females with both types of antibody was observed in families of carriers. Anti-HBc determinations in conjunction with other HBV markers, provide a useful new tool for epidemiologic studies.