Eisuke Tanaka

A real-time quantitative polymerase chain reaction method for hepatitis B virus in patients with chronic hepatitis B treated with lamivudine

OBJECTIVES:During treatment of chronic hepatitis B with lamivudine, changes in the level of hepatitis B virus (HBV) DNA were investigated using a real-time polymerase chain reaction (PCR) method with a detection limit of 1.7 log copies/ml (50 copies/ml) to clarify its clinical significance, particularly the association between HBV DNA levels and the emergence of tyrosine-methionine-aspartate-aspartate (YMDD) mutants.METHODS:Twenty-four patients who had received lamivudine therapy for >1 yr were studied. HBV DNA levels were determined using transcription-mediated amplification for sera with >3.7 log genome equivalents/ml, the Roche Monitor kit for sera with ≥2.6 log copies/ml, and real-time PCR for sera with < 2.6 log copies/ml (the detection limit was 1.7 log copies/ml). Patients were classified into three groups according to the minimal HBV DNA level attained during lamivudine therapy: the <1.7 log copies/ml group (eight patients), the 1.7–2.5 log copies/ml group (five patients), and the ≥2.6 log copies/ml group (11 patients).RESULTS:Pretreatment HBV DNA levels were significantly lower in the <1.7 copies/ml group than in the other two groups (p < 0.05). Neither the emergence of YMDD mutants nor a virological breakthrough of serum HBV DNA was observed in any of the eight patients in the <1.7 copies/ml group. In contrast, in the 1.7–2.5 copies/ml and ≥2.6 copies/ml groups, virological breakthroughs resulting from the emergence of YMDD mutants were observed in two of five patients and in all 11 patients, respectively (p < 0.001). Virological breakthroughs were observed at a mean of 49.6 ± 18.4 wk in 11 the patients in the ≥2.6 copies/ml group and at wk 107 and 115 in two patients in the 1.7–2.5 copies/ml group.CONCLUSION:The real-time PCR method is useful for predicting the emergence of YMDD mutants and the estimated time of their emergence.

SERO-EPIDEMIOLOGIC STUDY OF HEPATITIS C VIRUS INFECTION IN FUKUOKA, JAPAN

Abstract: We conducted an epidemiological study of 509 residents of H town, Fukuoka, Japan, to investigate the high mortality rate from liver disease. Antibodies to hepatitis C virus (HCV) (anti-HCV) were detected in 120 residents (23.6%); HCV RNA in 91 (17.9%), and hepatitis B surface antigen (HBsAg) in 13 (2.6%). Multivariate logistic regression analyses showed that presence of anti-HCV, male gender, and history of liver disease were associated with the presence of liver dysfunction, and that age of more than 40 years and a particular district were associated with the presence of anti-HCV. HCV RNA was more frequently detected in anti-HCV-positive men than women (41, or 85.4% versus 50, or 69.4%) (P < 0.05). The incidence of liver dysfunction was significantly higher in HCV RNA-positive men than women (32, or 66.7% versus 22, or 30.6%) (P < 0.05). These findings suggest that: (1) HCV was correlated with the high mortality rate from liver diseases, (2) there were district-related differences in the incidence of HCV, and (3) the lower frequency of elimination of HCV from men may explain why they showed a high mortality from liver disease.

Interferon-γ brings additive anti-viral environment when combined with interferon-α in patients with chronic hepatitis C

Abstract T-cell hyporesponsiveness may lead to chronicity of hepatitis C virus (HCV) infection. We evaluated whether interferon (IFN)-γ injection can bring a Th1-dominant environment to patients with chronic hepatitis C. Seventeen patients with genotype 1b received natural IFN-α 5MU daily for the first 2 weeks and three times a week for the next 22 weeks followed by natural IFN-γ 1 MU daily for 2 weeks. In 4 of 17 patients (23.5%), alanine aminotransferase (ALT) was normalized and 3 of these 4 patients (75.0%) cleared HCV RNA. β2 microglobulin (BMG), neopterin and soluble (s) Fas increased with IFN-α and increased more with IFN-γ. Serum interleukin (IL)-12, CD4 and CD8 remained unchanged with IFN-α but increased after IFN-α was replaced by IFN-γ. IL-10 was not changed either with IFN-α or γ. Productions of IL-2, IFN-γ and tumor necrosis factor (TNF)-α by peripheral blood mononuclear cells did not change by IFN-α therapy, however, they were enhanced at the end of IFN-γ therapy. Productions of IL-2 and 4 were unaffected. These results show that some immune parameters become Th1-dominant by additional IFN-γ in patients with chronic hepatitis C. Combination of these two IFNs should be explored.

Adefovir Dipivoxil as a Treatment for Hepatic Failure Caused by Lamivudine-Resistant HBV Strains

Hepatitis B virus (HBV) infection is a serious worldwide problem because it is one of the major causes of cirrhosis and hepatocellular carcinoma in endemic areas. The number of people with chronic HBV infection is 350 million globally (1). Until recently, the therapeutic option for chronic HBV infection has been limited to interferon. Seroconversion from hepatitis B e antigen (HBeAg) to anti-hepatitis B e antibody (anti-HBe) occurs in up to about 40% of patients treated with interferon monotherapy (2–4). Recently, lamivudine, a nucleoside analogue, has become the main therapeutic option for chronic HBV infection. Some clinical trials showed that lamivudine suppressed HBV replication effectively and induced histological improvement (5–7). However, the prolonged therapy with lamivudine has been associated with the emergence of drug-resistant viruses with the mutations in the polymerase gene coding for the YMDD (tyrosine, methionine, aspartate, aspartate) motif (8, 9). The lamivudine-resistant HBV strains have been observed in 14 to 32% of patients undergoing a 1-year treatment regimen of 100 mg daily (6, 7). Several new nucleoside analogues are now under development. Adefovir dipivoxil, a prodrug of the acyclic deoxyadenosine monophosphate analogue adefovir, displays potent antiviral activity against HBV (10, 11) and an in vitro study demonstrated the antiviral activity to be against both wild-type and lamivudine-resistant strains of

Severe alcoholic hepatitis successfully treated by leukocytapheresis: a case report.

BACKGROUND The prognosis of severe alcoholic hepatitis is poor, and there is no established method for a cure. METHODS A 34-year-old man was admitted to Kurume University Hospital because of severe liver dysfunction due to excess alcohol intake. He was treated with prednisolone and two sessions of granulocyte and monocyte adsorption apheresis (GCAP) using an Adacolumn, which removes leukocytes--especially granulocytes and monocytes--from the peripheral blood. We evaluated the changes in the serum levels of interleukin-6, interleukin-8, tumor necrosis factor-alpha, and soluble intercellular adhesion molecule-1, as well as the conventional liver tests and peripheral white blood cell count. RESULTS Prednisolone was effective in the short term but resulted in an increase in C-reactive protein (CRP), peripheral leukocytes, and serum total bilirubin. GCAP performed on the 34th and 41st hospital days produced decreases in the white blood cell count, total bilirubin, and intercellular adhesion molecule-1. The patient survived, despite the expected poor prognosis on admission. CONCLUSIONS GCAP is recommended as a potential therapeutic option for severe alcoholic hepatitis.

Antibody response to inactivated hepatitis A vaccine

Abstract We examined the seroconversion rate and level of hepatitis A virus antibodies induced by inactivated hepatitis A vaccine. Inactivated hepatitis A vaccine was administered intramuscularly to 127 healthy antibody negative subjects at doses of either 0.5 and 1.0 μ g two or three times. Convalescent sera from 21 adults with acute hepatitis A were also assayed. Serum antibody titers were measured by enzyme-linked immunosorbent assay. Seroconversion occurred in all 127 subjects (43 children, 84 adults). Acquired antibody levels in children in the early stage were higher than those in adults. Protective titers appeared 6 weeks after initial inoculation in 126 of 127 subjects and were maintained for at least 3 years in a 11 subjects. Mean maximum titer in subjects inoculated twice and three times after the initial vaccination were 393 mIU ml −1 at 6 weeks and 2745 mIU ml −1 at 7 months, respectively. Titers at 6 and 12 months after the initial vaccination were significantly higher in subjects with the 1.0 μ g inoculation than with 0.5 μ g. Titers in females were significantly higher than in males. The mean titer after inoculation was 1.1% of the convalescent titer in patients with acute hepatitis A. Neither adverse clinical events nor abnormal laboratory tests were observed after vaccination. Hepatitis A virus antibodies appeared reliably after inoculation with inactivated vaccine, persisting for at least 3 years. Although seroconversion rate was not age- or sex-dependent, the acquired titer was higher in children and females than in elderly males.

Analysis of serum hepatitis A virus antibody response in different courses of hepatitis A virus infection

Changes in the serum hepatitis A virus anti-body (anti-HAV) response in patients with different clinical courses of HAV infection were examined using immune adherence hemagglutination (IAHA). Anti-HAV was detected 2–6 weeks after the onset of clinical symptoms in patients with the typical course of acute hepatitis A and 1–4 weeks after the onset in those with fulminant hepatitis A. Maximal anti-HAV titers were observed 8–20 weeks after the onset of clinical symptoms, and changes in anti-HAV were similar in the typical and the prolonged course of acute hepatitis A, but maximal antibody titers were higher in the prolonged course. Maximal anti-HAV titers in patients with subclinical HAV infection were significantly lower than titers in patients with the typical and prolonged courses of acute hepatitis A, and in those with fulminant hepatitis A. High titers of anti-HAV remained positive for at least 6 years after infection in patients with clinical infection and for at least 4 years in patients with subclinical infection on follow-up. These findings suggest that the maximum anti-HAV titer correlates with the clinical severity of HAV infection; knowledge of the antibody response should be useful for analyzing the pathogenesis of HAV infection.

Clinical usefulness of serum HBV DNA measurement with a non radioactive chemiluminescence method

Abstract Aim, clinical usefulness of a non radioactive HBV DNA measuring method was evaluated. Subjects and methods, the subjects were 8 anti-HBe positive chronic hepatitis B (CH-B) patients and five CH-B patients who were orally administered reverse transcriptase inhibitor (RTI). Serum HBV DNA and DNA-P were serially determined at 1.5–2 month intervals for 1 year in CH-B with anti-HBe positive patients and before, 2, 4, 8, and 12 weeks after RTI administration. Serum HBV DNA was determined by two different non radioactive method (Viraprobe HB Lumi, Quantiplex HBV DNA). Results, the HBV DNA levels measured with two methods were highly correlated (P