Kusuya Nishioka

Hepatitis C in Hospital Employees with Needlestick Injuries

Hepatitis virus infection has been a problem among hospital employees. Hepatitis B virus (HBV) infection is now controlled by hospital safety practices limiting exposure to blood and other body flu...

High throughput screening of 16 million serologically negative blood donors for hepatitis B virus, hepatitis C virus and human immunodeficiency virus type-1 by nucleic acid amplification testing with specific and sensitive multiplex reagent in Japan.

Nationwide nucleic acid amplification testing (NAT) for hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) of blood donated voluntarily after serological screening was implemented on July 1st 1999 for transfusion and plasma fractionation by the Japanese Red Cross blood transfusion services. From February 1st 2000, HBV, HCV and HIV-1 NAT screening of pools of 50 negative serologically screened donated blood was started and the results were reported within 1 day after blood donation. Systems were established for rapid shipment, electronic communication, automated specimen preparation, pooling and automated amplification and detection. At present, NAT screening is carried out within 1 day after donation. This report describes the blood screening system by NAT and the results obtained from over 16 million blood samples using simultaneous screening for HBV, HCV and HIV-1 with multiplex reagent. Between February 1, 2000 and December 31, 2002, 16012175 serologically negative units were tested by NAT. 308 units with Hepatitis B virus DNA (HBV DNA) were detected. The sensitivity of 50 pool NAT screening with input volume of 0.2 ml is significantly higher than that of highly sensitive HBsAg testing. 46 cases with HCV RNA and six cases with HIV-1 RNA were detected. These cases were not detected by HCV antibody and HIV-1 antibody screening. The false positive rate was 0.18%. The NAT system was developed from serological screening test negative non-remunerated voluntary donations. We supply blood products to medical organizations after screening by NAT for HBV, HCV and HIV-1 for transfusion and source plasma for fractionation. This is the first automated integrated system for prevention of transfusion transmitted HBV, HCV and HIV-1 infections, by NAT screening.

Hepatitis B NAT virus‐positive blood donors in the early and late stages of HBV infection: analyses of the window period and kinetics of HBV DNA

Background and Objectives  The Japanese Red Cross (JRC) carries out nucleic acid amplification testing (NAT) for hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus‐1 (HIV‐1) by using a multiplex (MPX) reagent. Screening is undertaken on serologically negative units. In this study we characterized HBV NAT‐positive donations individually and analysed the window period and kinetics of HBV DNA, during acute infection, in follow‐up studies.

Glycyrrhizin inhibits the lytic pathway of complement - possible mechanism of its anti-inflammatory effect on liver cells in viral hepatitis -.

Glycyrrhizin, an aqueous extract of licorice root, has anti‐inflammatory activity and has been used for the treatment of chronic viral hepatitis. In the present study we describe the mechanism by which glycyrrhizin inhibits complement. Glycyrrhizin inhibited the cytolytic activity of complement via the activation of both the classical and alternative pathways, while it had no effect on immune adherence, suggesting that it blocks C5 or a later stage of the complement cascade. Further analysis revealed that glycyrrhizin inhibits the lytic pathway in which the membrane attack complex (MAC) is formed. This mechanism suggests that glycyrrhizin may prevent tissue injury caused by MAC not only in chronic hepatitis but in many autoimmune and inflammatory diseases.

Predictive value of screening tests for persistent hepatitis C virus infection evidenced by viraemia : Japanese experience

In November 1989, Japanese Red Cross Blood Centres started screening for heaptitis C virus (HCV) with enzyme‐linked immunosorbent assay (Elisa) for the C100‐3 viral peptide as the first such nationwide programme in the world. Thereafter post‐transfusion non‐A non‐B hepatitis (PTNANBH) was reduced by 61–80%, but this was not as complete a success as our programme to prevent post‐transfusion hepatitis B by screening for high titer hepatitis B core antibody, which we began in the same period. In order to acquire more effective control of PTNANBH, the HCV core‐related antigen (GOR, N14) and second‐generation Elisa (Ortho2, Abbott2)and second‐generation antigen agglutination (PA, PHA) tests have been employed. Among 16,500 donors in 11 blood centers, 365 were serologically positive by at least one of these tests. Among these, HCV RNA was detected in 138 units and the remaining 227 were HCV RNA negatives. The effectiveness of these serological tests to detect HCV RNA‐positive status were analyzed. Passive haemagglutination and particle agglutination (PHA and PA) tests were highly effective to predict HCV viraemia among blood donors. Also, these tests can easily determine antibody titre. By either PHA or PA, all units with ≧212 agglutination titre (120 and 122 units) were HCV RNA positive and all agglutination‐positive units with serum alanine aminotransferase level higher than 35 Karmen units were HCV RNA positive. These results have formed the basis for implementing a more effective screening for HCV viraemia in blood donors, where effectiveness is defined as enhancing the protection of patients from post‐transfusion hepatitis C and providing higher quality information to achieve more effective donor counselling.

Physiologic role of the complement system in host defense, disease, and malnutrition.

The role of the complement system as a system merging early-phase innate immunity with later-phase acquired immunity has been established. C3 is a key protein of the complement system. It is activated in four pathways: (1) the alternative pathway, (2) the mannan binding protein pathway, (3) the C-reactive protein pathway, and (4) the natural IgM pathway in innate immunity. It is also activated in (1) a classic pathway, i.e., through an antigen-antibody complex, and (2) by injured host cells in acquired immunity. Activation of C3 results in a variety of immunologic reactions such as immune adherence, phagocytosis, antibody response, cytolysis, inflammation, and killing of pathogenic microorganisms. Pathologic pictures of the complement system in various diseases were reviewed. Attention was focused on hypocomplementemia in the malnourished state. In humans and in experimental animals, reduced complement levels, especially of C3, were observed in relation to lowered host defense against infection. Hypocomplementemia improved after nutritional rehabilitation with a concomitant improvement of the clinical picture and recovery of host resistance. Enhancement of C3 levels in malnourished or well-nourished rats resulted in heightened resistance against bacterial infections. On the basis of these experimental and clinical observations, we obtained clues to prevent or treat a compromised host defense system in malnourished states.

Anti-hepatitis C virus antibody prevails in fulminant hepatic failure

SummarySerial serum samples obtained from 27 patients with fulminant hepatic failure (FHF) in a variety of etiology were tested for anti-hepatitis C virus antibody (anti-HCV). Seven out of 10 patients (70%) with FHF due to hepatitis B virus (HBV) infection were positive for anti-HCV, showing a significantly higher rate than that in acute HBV hepatitis (0/17, 0%): In particular, all 3 post-transfusion HBV-FHF cases were found to be positive for the antibody. The incidence of anti-HCV in sporadic non-A non-B (NANB)-FHF patients (7/11, 64%) tended to be greater than that in acute sporadic NANB hepatitis as recently surveyed in this country. In addition, anti-HCV was also detected in a patient with hepatitis A virus (HAV)-FHF and in 2 out of 4 drug-induced FHF patients. Moreover, anti-HCV appeared earlier in FHF (median; 27.5 days, n=9) regardless of etiology, when compared to acute NANB hepatitis (3 to 6 months). Hence, co-infection and/or superinfection of HCV with enhanced antibody response may play an important role in the development of this fatal disease.

The first large-scale nucleic acid amplification testing (NAT) of donated blood using multiplex reagent for simultaneous detection of HBV, HCV, and HIV-1 and significance of NAT for HBV.

The first nationwide nucleic acid amplification testing (NAT) for hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV‐1) of voluntarily donated blood after serological pre‐screening and before release of cellular components and plasma for fractionation was implemented by the Japanese Red Cross Blood Transfusion Services. From February 1, 2000 to April 30, 2001, specimens from 6,805,010 units of serologically negative donation were screened in minipools of 50 samples within 24 hr after blood donation by NAT using multiplex HBV/HCV/HIV‐1 reagent for blood transfusion including short shelf‐life platelets. Among them, 112 HBV DNA‐positives, 25 HCV RNA positives and 4 HIV‐1 RNA positives were screened out and we could prevent transfusion of these NAT positive units. Subtypes/genotypes of HBV DNA, adr/C, adw/A, adw/B, adw/C, ayr/C and ayw/D were found and adr/C was predominant. A total of 61.6% of them (69/112) were negative by overnight EIA. Sixth three of HBV NAT‐positive samples carried virus loads less than 104 copies/mL and 92.1% of them (58/63) were negative by overnight EIA. The virus growth curves of HBV in 6 cases obtained by retrospective and prospective follow‐up study showed exponential straight lines in the early stage of serological window periods and the log times of HBV growth (10 fold increase) in serological window period were between 4.6 and 7.6 days. NAT screening with highly sensitive reagents in pool of specimens is useful to exclude blood units with low level of HBV and HBV mutants from blood transfusion.

Detection and quantitation of HBV DNA by semi-nested PCR in donated blood: comparison with HBV serological markers

To detect and quantitate hepatitis B virus (HBV) DNA, semi-nested polymerase chain reaction (PCR) method was designed for amplifying the HBV core region DNA. Cloned HBV core region DNA was used as a quantitation control, and upon electrophoresis of the semi-nested PCR product, one, two, or three bands of amplified DNA were observed using a small (< 50 mol), moderate (around 200 mol), or large (> or = 1250 mol) quantity of the template DNA, respectively. Using this semi-nested PCR method, HBV DNA was quantitated in donated blood and tested for HBV serological markers. Most of the HBV surface antigen (HBsAg) high titer samples showed three bands on the electrophoresis, indicating a high level of HBV DNA, while most of the HBsAg low titer samples showed one band, indicating a low level of HBV DNA. HBV DNA was detected in 7 out of 36 HBsAg-undetectable and anti-HBc-positive samples (19.4%) but all 7 showed one band, indicating a low level of HBV DNA. In almost all of the HBV e antigen-positive samples the HBsAg titer was high, and three bands were observed indicating a high level of HBV DNA.

Hepatitis C virus infection in Japan.

SummaryBecause of the high incidence of non-A non-B post transfusion hepatitis and its relation to hepatocellular carcinoma, HCV antibody screening for blood transfusion was begun in November 1989. Seroepidemiological studies revealed the great magnitude of HCV infection in acute hepatitis, chronic hepatitis, liver cirrhosis and hepatocellular carcinoma in Japan. Approximately 1.6 million HCV antibody-positive persons are now in Japan. Major routes of infection are considered to be horizontal blood borne infection in the past and blood transfusion.