Masakazu Kobayashi

Anti‐gp210 and anti‐centromere antibodies are different risk factors for the progression of primary biliary cirrhosis

The predictive role of antinuclear antibodies (ANAs) remains elusive in the long‐term outcome of primary biliary cirrhosis (PBC). The progression of PBC was evaluated in association with ANAs using stepwise Cox proportional hazard regression and an unconditional stepwise logistic regression model based on the data of 276 biopsy‐proven, definite PBC patients who have been registered to the National Hospital Organization Study Group for Liver Disease in Japan (NHOSLJ). When death of hepatic failure/liver transplantation (LT) was defined as an end‐point, positive anti‐gp210 antibodies (Hazard ratio (HR) = 6.742, 95% confidence interval (CI): 2.408, 18.877), the late stage (Scheuers stage 3, 4) (HR = 4.285, 95% CI:1.682,10.913) and male sex (HR = 3.266, 95% CI: 1.321,8.075) were significant risk factors at the time of initial liver biopsy. When clinical progression to death of hepatic failure/LT (i.e., hepatic failure type progression) or to the development of esophageal varices or hepatocellular carcinoma without developing jaundice (Total bilirubin < 1.5 mg/dL) (i.e., portal hypertension type progression) was defined as an end‐point in the early stage (Scheuers stage 1, 2) PBC patients, positive anti‐gp210 antibodies was a significant risk factor for hepatic failure type progression [odds ratio (OR) = 33.777, 95% CI: 5.930, 636.745], whereas positive anti‐centromere antibodies was a significant risk factor for portal hypertension type progression (OR = 4.202, 95% CI: 1.307, 14.763). Histologically, positive anti‐gp210 antibodies was most significantly associated with more severe interface hepatitis and lobular inflammation, whereas positive anticentromere antibodies was most significantly associated with more severe ductular reaction. Conclusion: These results indicate 2 different progression types in PBC, hepatic failure type and portal hypertension type progression, which may be represented by positive‐anti‐gp210 and positive‐anticentromere antibodies, respectively. (HEPATOLOGY 2007;45:118–127.)

Antibody response to E2/NS1 hepatitis C virus protein in patients with acute hepatitis C

Antibody response to the E2/NS1 protein of the hepatitis C virus (HCV) was studied in 26 patients with post‐transfusion acute hepatitis C. Second‐generation HCV (HCV‐2) antibody, E2/NS1 antibody and HCV‐RNA were measured in serial serum samples taken within 1 month and 3, 6 and 12 months after the onset of acute hepatitis C. The HCV genotype was also tested to study its clinical significance. Of 26 patients, eight showed normalization of alanine aminotransferase (ALT) and clearance of HCV‐RNA (resolved group). In the remaining 18 patients, HCV viraemia and ALT abnormality (except one patient) continued for more than 3 years (unresolved group). Both HCV‐2 and E2/NS1 antibodies were positive at least once in all patients. The prevalence of E2/NS1 antibody was significantly (P < 0.05) higher in the resolved group (88%) than in the unresolved group (39%) in the period within 1 month of onset; the prevalence was similar between the two groups thereafter. The prevalence of HCV‐2 antibody did not differ between the two groups at any point. The HCV genotype was not related to the chronicity of acute hepatitis C. In conclusion, the E2/NS1 antibody appeared in all patients with acute hepatitis C and was associated with the clearance of HCV.

Detection of GB virus-C/hepatitis G virus genome in peripheral blood mononuclear cells and liver tissue.

The replication site for the GB virus‐C/hepatitis G virus (GBV‐C/HGV) was investigated by using polymerase chain reaction (PCR)‐based assays and in situ hybridisation. A total of 28 patients with consecutive GBV‐C/HGV infection were enrolled in this study: Nine patients were being treated with immunosuppressive therapy after liver transplantation, and the remaining 19 patients were not receiving such treatment. GBV‐C/HGV RNA was detected by using reverse transcriptase‐polymerase chain reaction (RT‐PCR) and was quantitated by using competitive RT‐PCR in all patients. Positive and negative strands of GBV‐C/HGV RNA in liver tissue were detected with in situ hybridisation by using RNA probes that were specific for the GBV‐C/HGV genome. Concentrations of GBV‐C/HGV RNA in serum were significantly higher (P = 0.003) in the nine patients who were receiving immunosuppression (median, 107 copy/ml; range, 105–107) than in the 19 patients who were not receiving immunosuppressive therapy (median, 104 copy/ml; range, 102–107). In situ hybridisation of GBV‐C/HGV RNA was performed on paraffin‐embedded liver tissue that was obtained from six patients with GBV‐C/HGV infection. Two of those six patients were receiving immunosuppressive therapy, and four were not. Significant positive signals were observed in the samples from two of the six patients who were infected with GBV‐C/HGV, but such signals were not observed in any of the six patients who were without the infection. The two patients with positive signals (both were undergoing immunosuppressive therapy) showed both positive and negative strands of GBV‐C/HGV RNA in mononuclear cells that infiltrated into portal areas, but neither of the strands was observed in hepatocytes. Moreover, the GBV‐C/HGV replication was analysed in peripheral blood mononuclear cells by using strand‐specific PCR (conventional RT‐PCR and rTth method). Two of the six patients were positive for negative‐strand GBV‐C/HGV RNA by using conventional RT‐PCR. In conclusion, GBV‐C/HGV replication was active under an immunosuppressive state, and it is suggested that GBV‐C/HGV replicates in mononuclear cells. J. Med. Virol. 57:114–121, 1999.

Quantitative measurement of HCV RNA in the serum: A comparison of three assays based on different principles

Quantitative measurement of hepatitis C virus (HCV) RNA is useful in patients with chronic hepatitis C, especially with interferon treatment. We examined the clinical usefulness of the AMPLICOR monitor assay, a newly developed assay for quantitative measurement, by comparing it with two other assays with different principles. A total of 48 patients with chronic hepatitis C who were treated with interferon‐α (IFN‐α) were studied: 19 were complete responders and 29 were non‐responders. Hepatitis C virus RNA was measured quantitatively by AMPLICOR, branched DNA (bDNA) probe, and competitive polymerase chain reaction (C‐PCR) assays. An internal quantification standard was used in the AMPLICOR assay. A cDNA competitor with a deletion of 15 base pairs in the middle portion was used in the C‐PCR method. The concentration of HCV RNA was significantly correlated between the three assays adopted in this study. Sensitivity of assays was 100% by C‐PCR, 90% by AMPLICOR and 69% by bDNA assays. The active quantitative range was best with the C‐PCR assay and worst with the bDNA assay. The bDNA assay had a tendency to exhibit lower values for patients with serotype 2 than did the other two assays. The predictive rate of the long‐term response to IFN‐α therapy, before its initiation, was over 75% in all three assays. The predictive rate just after completing IFN‐α therapy was as high as 80% by C‐PCR and the AMPLICOR assays, but was low (58%) with the bDNA assay. The handling of the bDNA and AMPLICOR assays was much easier than the C‐PCR assay, which required time and skill. These results indicate that the AMPLICOR assay is a simple and reliable method for measuring the serum concentrations of HCV RNA, and thus is suitable for clinical application.

Clinical application of hepatitis C virus core protein in early diagnosis of acute hepatitis C

Abstract: A fluorescence enzyme immunoassay (FEIA) for the quantitative measurement of hepatitis C virus (HCV) core protein has recently been developed. In this study, we studied the clinical usefulness of this measurement in patients with acute hepatitis C. Eighteen patients with post-transfusion acute hepatitis C were enrolled in the study; 5 patients showed resolution of hepatitis with disappearance of HCV viremia, while the remaining 13 patients did not. A second generation HCV antibody, HCV RNA, and HCV core protein were measured in serial serum samples taken within 1 month of the onset of acute hepatitis and 3, 6, 12, 24, and 36 months after onset. Within the first month after disese onset, the positivity rates of HCV RNA (100%; P = 0.0014) and HCV core protein (89%; P = 0.0300) were both significantly higher than that of HCV antibody (56%). Six months after disease onset, the positivity rate of HCV antibody had increased, to 100%, and the pasitivity rates of HCV RNA and HCV core protein began to decrease. HCV core protein levels did not differ between patients with resolved and unresolved disease in the first month after disease onset. These findings indicate that FEIA, a simple assay, for the measurement of HCV core protein was useful for the early diagnosis of acute hepatitis C.

A case of glycogen storage disease type Ia with multiple hepatic adenomas and G727T mutation in the glucose-6-phosphatase gene, and a comparison with other mutations previously reported.

We report a case of 23-yr-old man with glycogen storage disease (GSD) type Ia complicated by multiple hepatic adenomas. Analysis of the G-6-Pase gene using peripheral blood sample showed this patient to be homozygous for a G-to-T transversion at nucleotide 727 in exon 5. This mutation is prevalent among Japanese patients, suggesting that specific genotypes may correlate with different clinical courses or outcomes.

Hepatitis G virus/GB virus C infection in patients with chronic non-B, non-C hepatitis

Abstract A new virus named hepatitis G virus or GB virus C (HGV/GBV-C) was recently identified as a causative agent for acute and chronic hepatitis. In this study, we tested HGV/eGBV-C RNA in serum in addition to hepatitis B virus (HBV) DNA and hepatitis C virus (HCV) RNA to study the causative agents involved in chronic non-B, non-C hepatitis. Of the 1089 patients with histological evidence of chronic viral hepatitis, 25 (2.3%) were negative for both hepatitis B surface antigen and second generation HCV antibody in serum. The 25 patients diagnosed as having chronic non-B, non-C hepatitis, were investigated in this study. HGV/GBV-C a RNA was detected by nested reverse transcription polymerase chain reaction using primers in 5′-untranslated, and the nonstructural (NS) 3 and NS5 regions. Of the 25 patients with chronic non-B, non-C hepatitis, four (16%) were positive for HGV/GBV-C RNA, only one (4%) was positive for HBV DNA, and none were positive for HCV RNA. Of the four patients with HGV/GBV-C RNA, two histologically had mild fibrosis, and the remaining two had cirrhosis. One patient with cirrhosis also had hepatocellular carcinoma, HBV DNA was positive in this patient. All three patients with only the HGV/GBV-C infection had a mild histological grade. In conclusion, HGV/GBV-C infection was involved in 16% of Japanese patients with chronic non-B, non-C hepatitis. Chronic hepatitis G seemed to exhibit mild hepatitis activity.

KIR3DL1-HLA-Bw4 combination and IL28B polymorphism predict response to Peg-IFN and ribavirin with and without telaprevir in chronic hepatitis C.

Natural killer cells play a key role in the immune control of viral infections. Killer immunoglobulin-like receptors (KIRs) regulate natural killer cell activation and inhibition through the recognition of their cognate HLA class I ligands. We assessed the predictive factors of a sustained virological response (SVR) in 200 Japanese patients with chronic genotype 1b hepatitis C who were treated with telaprevir (TVR), pegylated-interferon-α2b (PEG-IFN), and ribavirin (RBV) triple therapy (92 patients) or PEG-IFN/RBV therapy alone (108 patients). Sixteen KIR genotypes, HLA-A, -B and -C ligands, and an interleukin (IL) 28B polymorphism (rs8099917) were analyzed. We observed that triple therapy, white blood cell count, hemoglobin value, hepatitis C viral load, a rapid virological response (RVR), IL28B TT genotype, and KIR3DL1-HLA-Bw4 genotype were associated with an SVR. In multivariate regression analysis, we identified an RVR (P < 0.000001; odds ratio [OR] = 20.95), the IL28B TT genotype (P = 0.00014; OR = 5.53), and KIR3DL1-HLA-Bw4 (P = 0.004, OR = 3.42) as significant independent predictive factors of an SVR. In conclusion, IL28B and KIR3DL1/HLA-Bw4 are independent predictors of an SVR in Japanese patients infected with genotype 1b HCV receiving TVR/PEG-IFN/RBV or PEG-IFN/RBV therapy.

Long-term outcomes of add-on adefovir dipivoxil therapy to ongoing lamivudine in patients with lamivudine-resistant chronic hepatitis B.

Aim:  Add‐on adefovir dipivoxil (ADV) therapy has been a standard rescue treatment for patients with lamivudine (LAM)‐resistant chronic hepatitis B, but the overall benefits of long‐term add‐on ADV therapy are still limited. The aim of this study was to evaluate the long‐term efficiency of add‐on ADV treatment and to explore predictive factors associated with it.

Malignant insulinoma presenting a non-functioning metastatic liver tumor 14 years after resection of the primary tumor

Abstract: A 58-year-old woman who had undergone resection of insulinoma 14 year earlier visited our clinic complaining of abdominal discomfort. Computerized tomographic scan showed multiple liver tumors, and a diagnosis of metastatic tumor of malignant islet cell tumor was confirmed histologically. No oversecretion of hormones or hypoglycemic episode was observed on readmission. Thus, the insulinoma seemed to have transformed to a clinically non-functioning tumor. The patient was treated with transcatheter arterial embolization, resulting in clinical improvement with marked reduction in tumor size. Features of interest in this case included; (1) transformation to non-functioning metastatic liver tumor 14 years after resection of insulinoma, (2) the usefulness of transcatheter arterial embolization for multiple metastatic tumor of malignant islet cell tumor.