Tolerance to hepatitis B antigen: a hypothesis for its termination with 'immune-RNA'

Immunologic tolerance to hepatitis B antigen, evidenced by a lack of specific cellular and humoral immune response to HB_sAg, produces a chronic carrier state which serves as an epidemiologic reservoir for the transmission of viral hepatitis type B. It is proposed that cell-mediated immunity transferred with immune-RNA may serve to terminate immunologic tolerance to hepatitis B antigen and abolish the carrier state. The efficacy and safety of ‘immune-RNA’ administration to chronic carriers can be validated by leukocyte migration inhibition techniques $\text{in vitro}$ and in HB_sAg positive chimpanzees $\text{in vivo}$.     

A double-blind evaluation of transfer factor therapy of HBsAg-positive chronic aggressive hepatitis: preliminary report of efficacy.

A controlled, prospective, double-blind trial of transfer factor in chronic aggressive hepatitis was carried out. This report presents the results obtained from study of the nine HB_sAg-positive subjects who were included in the trial. Transfer factor was prepared from adults who had recovered from acute hepatitis B or from acute non-B viral hepatitis and was administered to five HB_sAg-positive subjects. Four HB_sAg-seropositive subjects received placebo. Percutaneous liver biopsy was performed at the beginning and at the conclusion of the 10-week study period; biochemical and clinical parameters were monitored throughout. Cell-mediated immunity was assessed at the beginning and at the end of the study period. Four of five transfer factor recipients showed moderate or marked improvement in hepatic histology, clinical status, and serum transaminase levels, while none of four placebo recipients demonstrated improvement. The difference in response between the two groups was significant (P = 0.048). No consistent changes in in vivo or in vitro correlates of cell-mediated immunity were demonstrated. No adverse effects were noted among transfer factor recipients. These data suggest that transfer factor may be of benefit in chronic aggressive hepatitis associated with HB_sAg. Additional studies appear to be indicated to delineate the mode of action of transfer factor as well as the role that such immunotherapy should play in the management of patients with this disorder.     

Passive Immunization against Exposure to Hepatitis B Virus in the Military: Potential and Possibilities

The value of standard γ-globulin with a low titer of antibody to hepatitis B surface antigen (anti-HB_s) vs hepatitis B immune globulin (HBIG) in prevention of icteric hepatitis B in the military is unclear. Although recent studies have shown a decrease in icteric hepatitis after administration of both types of γ-globulin in populations where acquisition of hepatitis B virus (HBV) is most likely the result of nonparenteral transmission, the data pertaining to parenteral exposure suggest that HBIG delays the incubation period of HBV and decreases the development of passive-active immunity. Since no studies have demonstrated efficacy of standard γ-globulin or HBIG in a drug-using population where multiple HBV exposures are likely, the results observed in most trials are not comparable to hepatitis B associated with drug abuse in the military. Therefore, before a recommendation for use of routine γ-globulin or HBIG can be made for drug-related hepatitis in the military, efficacy of standard γ-globulin and/or HBIG should be demonstrated in this population.     

Hepatitis B surface antigen (HBsAg) infection in a hemodialysis unit. II. Factors affecting host immune response to HBsAg.

Serum from 86 hemodialysis patients, 105 healthy hospital staff "at risk" and 160 regular hospital staff was screened for hepatitis B surface antigen (HBsAg) and antibody (anti-HBs). The combined prevalence of HBsAg and anti-HBs was higher in the staff of the artificial kidney unit (57.7%) than in the hemodialysis patients (33.7%). The healthy subjects with HBsAg infection responded significantly more often by producing anti-HBs compared with the hemodialysis patients. Twelve of 29 (41.4%) hemodialysis patients with HBsAg infection produced anti-HBs, while 17 (58.6%) remained positive for HBsAg. This differential response could not be attributed to age, sex, time spent undergoing hemodialysis, delayed cutaneous reactivity or response to phytohemagglutinin (PHA) or pokeweed mitogen (PWM). However, a much larger proportion of patients with HBsAg than with anti-HBs had previously received blood transfusions (88.2% v. 33.3%). Our results indicate that development of the chronic HBsAg carrier state or production of anti-HBs in uremic patients may be influenced by the route of immunization or the dose of antigen, or both. Although uremic patients maintain normal in vitro response to PHA and PWM, they may have depressed immunity in vivo because of a decreased total number of T-lymphocytes.     

Viral Hepatitis: Problems of Incidence and Control in Military Personnel

(A) Hepatitis B virus (HBV) is now the major cause of infectious viral hepatitis in U.S. military personnel and probably also in the civilian population over 15 years of age. (B) The incidence of icteric, viral hepatitis is much higher in U.S. military personnel than in comparable age groups in the civilian population. The 17-to 20-year-old enlisted men show the highest rates. (C) In parts of the world (e.g., U.S.A., Germany) where most of the inapparent infection is caused by the adw subtype of HBV, most of the acute clinical disease is caused by the ayw subtype. In the U.S.A. and Germany, 95% or more of HB_s Ag isolates from U.S. military personnel with acute hepatitis is ayw. (D) It may be many years before one can expect to have sufficient data for a decision as to the possible availability of an effective HBV vaccine. Accordingly, a decision is urgently needed regarding either the immediate use of the best practically available hepatitis immune gamma globulin, that can be prepared by modern techniques, for the prevention of hepatitis in U.S. military personnel or postponement of such use until an adequate and properly controlled trial can be carried out in active duty military personnel in an area of high incidence.     

Assessment of peripheral DNA damage by alkaline comet assay in maintenance hemodialysis subjects with hepatitis C infection

Despite the high prevalence of hepatitis C infection among hemodialysis subjects, there is no information concerning the DNA damage of hepatitis C (+) hemodialysis subjects. We aimed to find out if there is any additional effect of hepatitis C infection on peripheral DNA damage in maintenance hemodialysis subjects. Fifteen hepatitis C (+) and 22 hepatitis C (-) hemodialysis subjects, 21 hepatitis C subjects without renal disease, and 22 healthy controls were enrolled. Peripheral DNA damage was assayed using alkaline comet assay. Median DNA damage levels of the study groups were as follows: hepatitis C (+) maintenance hemodialysis subjects, 88 (0-232); hepatitis C (-) maintenance hemodialysis subjects, 58 (0-228); hepatitis C (+) subjects without renal disease, 112 (44-252); controls, 26 (0-72). DNA damage level was significantly higher among hepatitis C (+) subjects without renal disease than hepatitis C (-) maintenance hemodialysis subjects and healthy controls (both p<0.05/6). Both maintenance hemodialysis subjects with and without HCV infection had significantly higher DNA damage level than healthy controls (both p<0.05/6). DNA damage level was comparable between hepatitis C (+) subjects without renal disease and HCV (+) hemodialysis subjects, and between hemodialysis subjects with and without hepatitis C infection (all p>0.05/6). Linear regression analysis revealed that hepatitis C infection was the only independent factor in predicting the peripheral DNA damage (p<0.05, beta=0.395). Each one of end-stage renal disease and hepatitis C infection significantly increases DNA damage level. However, in hemodialysis subjects, hepatitis C infection does not cause significant additional increase in DNA damage level, and it may be partly due to protective effect of hemodialysis on hepatitis C infection.     

Seroepidemiologic study of adult T-cell leukemia virus (ATLV) and hepatitis B virus infection in Okinawa, Japan

A total of 2,283 serum samples were collected from healthy subjects in three islands of the Yaeyama district of Okinawa, Japan. These sera were tested for the presence of hepatitis B surface antigen (HBsAg), for antibody to hepatitis B core antigen (anti-HBc), and for antibody to adult T-cell leukemia-associated antigen (anti-ATLA). Correlation between hepatitis B virus infection and adult T-cell leukemia virus (ATLV) infection was determined by using the prevalence rates for three virus markers. Overall prevalence of HBsAg, anti-HBc and anti-ATLA was 6.5%, 57.4%, and 17.9%, respectively. Age-specific prevalence of anti-HBc and anti-ATLA increased with age, but that of HBsAg did not. Sex-specific prevalence of HBsAg was significantly higher in males than in females, but that of anti-ATLA was significantly higher in females than in males. Statistical analysis revealed that prevalence of anti-ATLA was significantly higher in HBsAg-positive persons and HBsAg-negative/anti-HBc-positive persons than in those negative for HBsAg and anti-HBc. These data suggest that hepatitis B virus-infected persons have a significantly higher chance of adult T-cell leukemia virus infection than those without hepatitis B virus infection in the area studied.     

General Epidemiological Parameters of Viral Hepatitis A,B, C,and E in Six Regions of China: A Cross-Sectional Study in 2007

Background

Viral hepatitis is a serious health burden worldwide. To date, few reports have addressed the prevalence of hepatitis A, B, C, and E in China. Therefore, the general epidemiological parameters of viral hepatitis remain unknown.

Principal Findings

In this cross-sectional study, we performed a serological prevalence analysis of viral hepatitis A, B, C, and E in 8,762 randomly selected Chinese subjects, which represented six areas of China. The overall prevalence of anti-Hepatitis C virus antibody (anti-HCV) was 0.58%, which was much lower than was estimated by WHO. The prevalences of Hepatitis B virus surface antigen (HBsAg), anti-Hepatitis B virus surface protein antibody (HBsAb), and anti-Hepatitis B virus core protein antibody (HBcAb) were 5.84%, 41.31%, and 35.92%, respectively, whereas in the group of subjects less than 5 years old, these prevalences were 1.16%, 46.77%, and 8.69% respectively, which suggests that the Hepatitis B virus (HBV)-carrier population is decreasing, and the nationwide HBV vaccine program has contributed to the lowered HBV prevalence in the younger generation in China. Meanwhile, a large deficit remains in coverage provided by the national HBV immune program. In addition, our data suggested the possibility that HBsAb may not last long enough to protect people from HBV infection throughout life. The overall prevalence of anti-Hepatitis A virus antibody (anti-HAV) and anti-Hepatitis E virus antibody (anti-HEV) were as high as 72.87% and 17.66%, respectively. The indices increased with age, which suggests that a large proportion of Chinese adults are protected by latent infection. Furthermore, the pattern of HEV infection was significantly different among ethnic groups in China.

Conclusions

Our study provided much important information concerning hepatitis A, B, C, and E prevalence in China and will contribute to worldwide oversight of viral hepatitis.     

Genotype and subtype analyses of hepatitis B virus (HBV) and possible co-infection of HBV and hepatitis C virus (HCV) or hepatitis D virus (HDV) in blood donors, patients with chronic liver disease and patients on hemodialysis in Surabaya, Indonesia

Four subtypes (adw, adr, ayw, and ayr ) and eight genotypes (A to H) of the hepatitis B virus (HBV) have been identified. They appear to be associated with particular geographic distribution, ethnicity, and possibly clinical outcomes. In this study, hepatitis B surface antigen (HBsAg) subtyping and HBV genotyping were carried out on sera obtained from HBsAg-positive HBV carriers, including healthy blood donors; patients with acute hepatitis, chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma; and patients on hemodialysis all located in Surabaya, Indonesia. We report here that all HBV isolates tested in Surabaya belonged to genotype B, with more than 90% of them being classified into subtype adw. Our results also revealed that prevalence of hepatitis C virus (HCV) co-infection among HBV carriers in Surabaya was approximately 10% for healthy blood donors and patients with chronic liver disease, and approximately 60% for patients on maintenance hemodialysis. Interestingly, HBsAg titers were lower in HBV carriers with HCV co-infection than in those without HCV co-infection. We also found that prevalence of hepatitis D virus (HDV) co-infection was < 0.5% among HBV carriers in Surabaya.     

Hepatitis B in a hospital for the mentally subnormal in southern England.

The prevalence of hepatitis B viral (HBV) infection was assessed in 340 patients and 268 staff in a hospital for the mentally subnormal in Wessex. Hepatitis B surface antigen (HBsAg), antibody to hepatitis B core antigen, antibody to HBsAg, e antigen (eAg), and antibody to eAg were used as markers of such infection. Forty patients and 10 staff had evidence of recent or current infection, while 149 patients and 50 staff had evidence of past infection. HBV markers were more common in mongols, epileptics, patients with cerebral palsy, and those of lower mental grades and reached a peak after 5-15 years of hospitalisation. eAg was detected in 12 out of 26 patients with HBsAg but in none of the four staff with HBsAg. Abnormal liver function values were found in 24 (60%) of the patients with recent or current HBV infection but in only 30 (19%) of those without HBV markers. Among the staff the prevalence of HBV markers correlated with the duration of employment and degree of contact with patients. Of those with recent or current infection, 4 (40%) had abnormal liver function values compared with 25 (12%) of those without HBV markers. Despite the high prevalence of markers clinically overt hepatitis B was rare.     

Vaccine against human hepatitis B virus prepared from antigen derived from human hepatoma cells in culture

Vaccine against human hepatitis B was prepared using antigen derived from hepatitis B carrier hepatoma cells grown in the interstices of a Diaflo hollow filter unit. Hepatitis B surface antigen (HBsAg) produced by these cells was purified by immune affinity chromatography, digestion with DNase and pepsin, and Sephadex G-150 separation. The Formalin-treated antigen was formulated in 20-micrograms dose on alum adjuvant with thimerosal added as a preservative. This cell culture vaccine was as potent as human plasma-derived vaccine as measured in a mouse potency assay. The vaccine proved safe in tests in chimpanzees and in human subjects who were in late stages of cancer of the central nervous system and who were receiving therapy for their condition. None of five subjects who received the vaccine developed untoward clinical reactions. Two of the subjects who received all three doses of vaccine developed antibody against HBsAg. Three persons, two given only the primary doses and one who was given all three doses but was lost to follow-up, demonstrated no response. The slow and relatively low antibody responses to the vaccine were similar to those in other immunosuppressed persons who were given vaccine of human plasma origin.     

Evaluation of an immunoenzyme test (ELISA) for the determination of an Ag/Ab system correlated to non-A, non-B post-transfusion hepatitis

An ELISA for the detection of a Ag/Ab system related to non-A, non-B post-transfusion hepatitis (NANB-PTH) has been developed. A convalescent serum from a patient suffering, 14 month before, of NANB-PTH was used both as capture and detector antibody in a "sandwich" type immunoassay to detect an Ag (Ag-NANB). A "blocking" immunoassay was used, in the same test, to detect the corresponding antibody (Ab-NANB). All patients with NANB-PTH were positive, alternately, for Ag-NANB or Ab-NANB. The Ag was found in the acute phase sera of 3 out of 22 cases of NANB-PTH (13,6%), but in none of 29 acute hepatitis type B,26 acute hepatitis type A,14 NANB sporadic hepatitis, 5 infectious mononucleosis and 8 healthy subjects. 2/3 Ag-NANB positive subjects became chronic carriers of this Ag, whereas the third seroconverted for Ab-NANB after the first two months of their disease. Furthermore, a significant increase of the Ab-NANB titer was observed in the convalescent phase of the disease in 18/20 NANB-PTH. As expected, a relatively high prevalence for Ab-NANB was found in the other groups of patients, suggesting the high diffusion of the agent, but none of these subjects showed a progressive increase of the antibody titer during the course of their disease.     

Hepatitis A and B: serologic survey of human and nonhuman primate sera

Sera of humans and seven species of nonhuman primates were tested by radioimmunoassay and enzyme immunoassay for the presence of hepatitis A antibody, hepatitis B surface antigen and antibody to hepatitis B surface antigen. The outcome of testing a total of 276 serum or plasma specimens was as follows: with the exception of squirrel monkeys (0%) and cotton-top marmosets (0%), a considerable percentage of all other species tested had detectable antibodies to hepatitis A virus: humans 45.9%, chimpanzees 36.6%, baboons 38.2%, vervets 57.9%, cebus monkeys 40.0% and common marmosets 50.0%. Only one human and two chimpanzees were carriers of hepatitis B surface antigen. Antibodies to hepatitis B surface antigen were detected in human (11.3%), chimpanzees (29.9%), baboons (36.2%) and squirrel monkeys (5%). Chimpanzees showed an increasing prevalence of antibodies to hepatitis A virus and hepatitis B surface antigen with age.     

Prevalence of hepatitis B viral markers in Vietnamese blood donors

Serum samples were assayed using radioimmunoassay in 573 Vietnamese blood donors living in Hano? (North Viet Nam). 66 (11.5%) subjects were HBsAg-positive. Of these 66 HBsAg carriers, 17 (25,8%) were positive for hepatitis B e antigen (HBeAg) and 43 (65.1%) for antibody to HBeAg (anti-HBe). 22 (3.8%) subjects were positive for antibody to hepatitis B core antigen (anti-HBc) alone. 402 (70.2%) subjects were positive for antibody to HBsAg (anti-HBs). This anti-HBs percentage increased with age. Only 83 (14.5%) subjects were negative for all hepatitis B viral (HBV) markers. This no HBV markers percentage decreased with age. The chi 2 test showed a non significant difference for frequencies of HBsAg, anti-HBc alone, anti-HBs but a significant one for frequencies of no HBV markers in men and women.     

A microtiter solid-phase radioimmunoassay for hepatitis A antigen and antibody.

A microtiter solid phase radioimmunoassy for hepatitis A antigen (HA Ag) and antibody (anti-HA) was developed. The test was more sensitive than immune adherence hemagglutination for detecting HA Ag and almost as sensitive for detecting anti-HA. The specificity and sensitivity of reagents were examined and optimum conditions for the test were determined. Radioimmunoassay, immune adherence hemagglutination, and immune electron microscopy were compared for detecting anti-HA. A serologic response to HA Ag was detected in paired sera from patients with type A hepatitis but not from patients with type B or non-A, non-B hepatitis by all three techniques.     

Serologic markers for hepatitis B among Marshallese accidentally exposed to fallout radiation in 1954

At least one serologic marker of prior hepatitis B infection (hepatitis B surface antigen, antibody to surface antigen, or antibody to core antigen) was found in 91.7% of 314 Marshallese tested. The prevalence of hepatitis B surface antigenemia (3.3%) in a subpopulation that had resided on Rongelap Atoll at the time of accidental exposure to radioactive fallout from a thermonuclear test in 1954 did not differ significantly from the prevalence in a selected unexposed population (10.5%).     

Hepatitis A antigen isolated from liver and stool: immunologic comparison of antisera prepared in guinea pigs.

Morphologically similar hepatitis A antigen particles (HA Ag)3 have been detected in the stools of patients with type A hepatitis and in the livers of marmosets experimentally infected with hepatitis A virus. To investigate the humoral antibody responses to these antigens and to compare the immunologic properties of HA Ag from these two sources, we immunized guinea pigs with either marmoset liver-derived HA Ag or with human stool-derived HA Ag in complete Freund's adjuvant and measured their antibody responses by immune electron microscopy (IEM) and immune adherence hemagglutination (IAHA). Antibodies reacting with both hepatitis A antigens were elicited in both groups. As determined by IEM, no distinction was seen between the reaction of guinea pig antiserum to each HA Ag tested under code when reacted against either liver-derived or stool-derived HA Ag. Antibodies elicited to marmoset liver-derived HA Ag and human stool-derived HA Ag had similar end point dilution titers by IAHA when tested against either "light" density (1.34 g/cm3) or "heavy" density (1.40 g/cm3) stool-derived HA Ag or liver-derived HA Ag. Low levels of antibody to normal liver or stool control antigens were observed transiently but did not obscure the specific response to HA Ag. These data suggest that morphologically similar HA Ag particles from different sources and with different densities are immunologically similar and may be identical. In contrast to the heterogeneity of surface antigens of hepatitis B virus, the comparable immunogenicity and apparent antigenic homogeneity of HA Ags derived from various sources may simplify the approach to development of a vaccine against viral hepatitis, type A.     

Immune-indian ink method for detection of hepatitis A associated antigen and antibody.

TIndian-ink grains coated with commercial gamma globulin (immune-Indian-ink) were agglutinated by 3 percent of sera from healthy volunteer blood donors; by 4 percent of those from hospital staff in contact with patients suffering from hepatitis; and by 10 percent of those from patients with viral diseases other than hepatitis, In contrast, the rate of positive reactions was 86 percent in the case of sera taken from patients in the acute phase of an illness diagnosed as hepatitis A on the basis of epidemiological and clinical data. Investigation of serum samples taken serially from patients positive in the acute phase of illness revealed that the immune-Indian-ink agglutinating factor does not persist for long in majority of cases. Two months after discharge from the hospital it was present in 18 percent of the patients only. The reaction proved negative when a limited number of cases diagnosed as hepatitis B were investigated. The immune-Indian-ink agglutinating factor was inhibited by all but one of 36 sera taken in the convalescent phase from patients with a diagnosis of hepatitis A. Some sera displaying agglutination with immune-Indian-ink gave a reaction with uncoated Indian-ink, too. Efforts to free the sera from non-specific agglutinating factor by starch-block electrophoresis have led to partial success. Fractionation on Sephadex G-200 columns suggested that in molecular weight (or particle size) the immune-Indian-ink agglutinating factor is smaller than HBsAg and larger than the non-specific agglutinating factor. On the basis of these results it is assumed that the immune-tindian-ink reaction is suitable for detecting an antigen tentatively called IH chi Ag and its antibody (IH chi Ab) specific to hepatitis A.     

Hepatitis B and C in the hemodialysis unit of Tocantins,Brazil: serological and molecular profiles

A survey was conducted in the hemodialysis population of the state of Tocantins, Brazil, aiming to assess the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections, to analyze associated risk factors, and also to investigate these viruses genotypes distribution. During January and March 2001, all patients (n = 100) were interviewed at the unique dialysis unit in Tocantins. Blood samples were collected and serum samples were screened for HBV serological markers. Hepatitis B surface antigen positive samples were tested for HBV DNA. All samples were also tested for anti-HCV antibodies and HCV RNA. An overall prevalence of 45% was found for HBV infection (4% were HBsAg/anti-HBc positive, 2% were anti-HBc only and 39% had anti-HBc/anti-HBs markers). Concerning HCV infection, anti-HCV and HCV RNA were detected in 13% and 14% of the subjects, respectively. Three patients were HCV RNA positive and anti-HCV negative, resulting in an overall HCV prevalence of 16%. Univariate analysis of risk factors showed that only shift and length of tile on hemodialysis were associated with HBV and HCV positivity respectively. Among the four HBsAg-positive samples, HBV DNA was detected in three of them, which were identified as genotype A by restriction fragment length polymorphism (RFLP) analysis. All 14HCV RNA-positive samples were genotyped by INNO-LiPA. Genotypes la and 3a were found in 85% and 15%, respectively. The present data show low HBsAg and HCV prevalence rates. The risk factors associated with HBV and HCV positivity suggest that nosocomial transmission may influence in spreading these viruses in the dialysis unit studied.