Kiyoshi Furuta

Transmission of hepatitis C in an isolated area in Japan : community-acquired infection

Abstract Background/Aims: The spread of hepatitis C virus (HCV) infection not due to drug needle sharing or transfusion is largely unknown in communities. A search for risk factors for HCV infection in an endemic area might elucidate inapparent modes of transmission. Methods: We conducted screening for hepatitis virus markers and parenteral exposures to blood among 435 inhabitants in an isolated area known for its endemicity for non-A, non-B hepatitis and in a nonendemic area with 1542 inhabitants. Results: The prevalence of hepatitis B surface antigen was the same in both areas. The prevalence of antibody to HCV verified by the recombinant immunoblot assay was 32.4% in the highly endemic area and 2.3% in the nonendemic area ( P Conclusions: Folk remedies such as acupuncture and cutting of the skin using nonsterilized knives should be considered as possible routes of HCV transmission not associated with blood transfusion or sharing of drug paraphernalia.

Fulminant hepatitis after allogenic bone marrow transplantation caused by reactivation of hepatitis B virus with gene mutations in the core promotor region

Abstract:  Under immunosuppressive conditions after hematopoietic stem cell transplantation (HSCT), even if hepatitis B virus (HBV) antigen is negative but hepatitis B surface antibody (HBsAb) or hepatitis B core antibody (HBcAb) is presented, HBV reactivates and sometimes causes fulminant hepatitis. However, it remains unclear which patients will develop fulminant hepatitis, or whether fulminant hepatitis is caused by host‐related factors or by virus‐related factors. A 30‐yr‐old man with a history of aplastic anemia since 3 yr of age underwent allogenic BMT, when HBsAb and HBcAb were positive but HBs antigen (HBsAg) was negative. The donor was negative for HBsAg, HBsAb and HBcAb. After transplantation, the patient was complicated by acute graft‐vs.‐host disease (GVHD), cytomegalovirus infection, intestinal thrombotic microangiopathy and aspergillus colitis. Chronic GVHD was well controlled by FK506 and prednisolone. Twenty months after transplantation, the patient was admitted with general fatigue and liver dysfunction and was found to be positive for HBsAg and HBeAg. His serum HBV‐DNA level was >8.8 log of the genome equivalent (LGE)/mL. Therefore, he was diagnosed as having hepatitis B caused by HBV reactivation and 100 mg/d lamivudine treatment was started. However, jaundice and hepatic failure deteriorated and became fatal. On analysis of the HBV‐DNA, two adjacent gene mutations in the core promoter region (T1762/A1764) were detected. Increased replication of the mutated HBV might have caused HBV reactivation which progressed to fulminant hepatitis.

A Case of Granulocyte-Colony Stimulating Factor-Producing Hepatocellular Carcinoma Confirmed by Immunohistochemistry

Granulocyte-colony stimulating factor (G-CSF) is a naturally occurring glycoprotein that stimulates the proliferation and maturation of precursor cells in the bone marrow into fully differentiated neutrophils. Several reports of G-CSF-producing malignant tumors have been published, but scarcely any in the hepatobiliary system, such as in hepatocellular carcinoma (HCC). Here, we encountered a 69-yr-old man with a hepatic tumor who had received right hepatic resection. He showed leukocytosis of 25,450/µL along with elevated serum G-CSF. Histological examination of surgical samples demonstrated immunohistochemical staining for G-CSF, but not for G-CSF receptor. The patient survived without recurrence for four years, but ultimately passed away with multiple bone metastases. In light of the above, clinicians may consider G-CSF-producing HCC when encountering patients with leukocytosis and a hepatic tumor. More cases are needed to clarify the clinical picture of G-CSF-producing HCC.

Genome-wide association studies identify PRKCB as a novel genetic susceptibility locus for primary biliary cholangitis in the Japanese population

&NA; A previous genome‐wide association study (GWAS) performed in 963 Japanese individuals (487 primary biliary cholangitis [PBC] cases and 476 healthy controls) identified TNFSF15 (rs4979462) and POU2AF1 (rs4938534) as strong susceptibility loci for PBC. In this study, we performed GWAS in additional 1,923 Japanese individuals (894 PBC cases and 1,029 healthy controls), and combined the results with the previous data. This GWAS, together with a subsequent replication study in an independent set of 7,024 Japanese individuals (512 PBC cases and 6,512 healthy controls), identified PRKCB (rs7404928) as a novel susceptibility locus for PBC (odds ratio [OR] = 1.26, P = 4.13 × 10‐9). Furthermore, a primary functional variant of PRKCB (rs35015313) was identified by genotype imputation using a phased panel of 1,070 Japanese individuals from a prospective, general population cohort study and subsequent in vitro functional analyses. These results may lead to improved understanding of the disease pathways involved in PBC, forming a basis for prevention of PBC and development of novel therapeutics.

Establishment of Epstein-Barr virus-associated lymphoma cell line SUBL with t(2;3)(p11;q27) from a liver transplant patient

A new lymphoma cell line, designated SUBL, was established from a Japanese patient with Epstein-Barr virus (EBV)-associated lymphoma, which developed during FK 506 therapy after liver transplantation. This cell line has undergone 80 passages over a period of 22 months. The cultured cells were positive for CD19, CD20, CD21, CD22, CD23, and HLA-DR, and negative for CD10 and surface immunoglobulins. Immunoglobulin gene analysis revealed rearrangements of JH and JK. T-cell antigens or T-cell receptor gene rearrangements were not observed on the cell line. The SUBL cells were positive for Epstein-Barr virus nuclear antigen (EBNA). The EBV genome was detected in the original tissue and the cell line by the in situ hybridization method. These data indicate that this cell line represents the B-cell lineage at a pre-B-cell stage. SUBL cells showed successful heterotransplantation to mice with severe combined immunodeficiency (SCID). Chromosomal analysis revealed the karyotype 46,XY,t(2;3)(p11;q27). Molecular studies showed that c-myc, N-myc, and bcl-2 were not rearranged. This cell line will provide a useful in vitro system to study the relationship between chromosomal abnormalities and the activation of cellular oncogenes.

Three-dimensional ultrastructure of normal rat hepatocytes by quick-freezing and deep-etching method

The three‐dimensional ultrastructure of nuclei and cell organelles including rough‐surfaced endoplasmic reticulum (RER), mitochondria, and cytoskeleton were studied in normal rat hepatocytes by the quick‐freezing and deep‐etching method. Peroxisomes and mitochondria were observed as spherical structures with granular matrices. Peroxisomes were identified by their size and matrices, which were more condensed than those of mitochondria. Ribosomes were identified as granular structures and were attached to the surface of endoplasmic reticulum. Cytoskeletal filaments were identified by their differences in diameter on the replica membranes, as well as in conventional ultrathin sections. Microfilaments were mainly localized around the bile canaliculi and adjacent to sinusoids. Intermediate filaments were observed around the bile canalicular microfilaments. Only a few filaments were observed near the lateral plasma membranes. Cross‐bridges measuring 5–7 nm in diameter were localized between the lamellae of RER and the surface of mitochondria. The quick‐freezing and deep‐etching method could be used to clarify the three‐dimensional association between the cytoskeleton and membrane‐bound organelles in hepatocytes.

Anti-c100 antibodies to hepatitis C virus in patients with chronic hepatitis C virus infection treated with interferon.

To assess the role of anti-c100 antibodies to hepatitis C virus (anti-c100 HCV) in the response to interferon (IFN) administered to patients for the treatment of chronic hepatitis C, we assayed serum anti-c100 HCV serially by means of an enzyme-linked immunosorbent assay (ELISA) kit and titrated anti-c100 HCV level by a radioimmunoassay (RIA) kit in 28 IFN-treated and 20 untreated patients with chronic hepatitis C. IFN-alpha or -beta was administered for 4 to 12 weeks, and the patients were observed for over 24 months. The IFN-treated patients were divided into 3 groups (4 sustained responders, 18 transient responders, and 6 non-responders) in accordance with their responses on the basis of the serum alanine aminotransferase levels. In three of the four sustained responders whose HCV-RNA decreased, anti-c100 HCV became negative at 13, 15, and 17 months after beginning the IFN therapy. The 18 transient responders and 6 non-responders remained positive for anti-c100 HCV throughout the observation period. In all four sustained responders the anti-c100 HCV titer decreased significantly with time after initiation of the therapy, whereas the titer fluctuated in the other groups. These findings suggest that monitoring the serum anti-c100 HCV level is useful in assessing the effects of IFN therapy on chronic hepatitis C.

Comparison of the clinical and immunogenetic features between patients with autoimmune hepatitis and patients with type C chronic active hepatitis

SummaryWe clarified the clinical and immunogenetical differences between patients with autoimmune hepatitis (AI-CAH), and patients with type C chronic active hepatitis (C-CAH) and type B chronic active hepatitis (B-CAH) who were positive for autoantibodies and hyperglobulinemia. While histories of blood transfusion, intravenous drug abuse and tattoo were seen frequently in patients with type C-CAH, they were rare in patients with AI-CAH. The severe subjective symptoms including anorexia, lethargy, icterus, high fever and extrahepatic manifestations, and severe abnormality of biochemical data were seen in AI-CAH predominantly. Ongoing or past infection of HCV was seen in only 14% of patients with AI-CAH. HLA-DR4 was the most frequently associated with AI-CAH (89%) and 6 DR4-negative patients were positive for DR2. HLA-DNA typing showed that there was no significant difference in the frequency of DR4-associated Dwalleles between the patients and controls who were positive for DR4. These findings suggest that the basic amino acid at position 13, which is present only on the DR2 and DR4 B1 molecules (Arg on DR2 and His on DR4), may contribute to the susceptibility to autoimmune hepatitis of Japanese. Thus, we conclude that AI-CAH is a genetically restricted, disease, and different from CCAH which is a viral infectious disease.

Ultrastructural studies of hepatocyte cytoskeleton in experimental cholestasis by quick-freezing and deep-etching method

The ultrastructural association between the cytoskeleton and other organelles was studied by the quick‐freezing and deep‐etching method in rats treated with α‐naphthylisothiocyanate (ANIT), or phalloidin, and in rats with obstructive jaundice. Cytoplasmic filaments were classified by measuring their diameters, and actin filaments were identified by specific decoration with myosin subfragment 1 (S1). S1‐positive actin filaments and S1‐negative intermediate filaments (12–14 nm in diameter) were observed to form a three‐dimensional network around bile canaliculi, and were more numerous than in controls, not only in phalloidin‐treated rats and rats with obstructive jaundice, but also in ANIT‐administered rats. In all cholestatic rats, vesicular structures were also more numerous than in controls in the pericanalicular regions, and were closely associated with the microfilaments and the intermediate filaments. Filaments of a new type were localized between the lamellae of rough‐surfaced endoplasmic reticulum and mitochondria, and between the lamellae of Golgi sacs and vesicles. Other thin filaments were also observed within the network of actin filaments. These filaments were 4–6 nm in diameter on replica membranes and were never decorated with S1. They were also directly connected with the canalicular membranes. Cytoskeletal components associated with membrane‐bound organelles, including these new filaments, were suggested to be involved in the localization and migration of organelles.

Serum auto‐antibodies in patients with hepatocellular carcinoma: The clinical significance of antinuclear antibody

Auto‐antibodies including antinuclear antibody (ANA), antismooth muscle antibody, antimitochondrial antibody, rheumatoid factor, lupus erythematosus factor, antimicrosomal antibody and antithyroglobulin antibody were assayed in sera from 84 patients with hepatocellular carcinoma, 70 with liver cirrhosis, 50 with chronic hepatitis, 30 with cancer of the alimentary tract and 100 normal subjects. A significantly higher incidence and higher titre of ANA was found in patients with hepatocellular carcinoma, and the patterns of nuclear fluorescence detected by the indirect immunofluorescent test using cultured baby hamster kidney cells were predominantly speckled and nucleolar. In 16 patients with this disease, serial assays of ANA were done in sera obtained before and after development of hepatoma and/or after resection of the hepatic tumour. Antinuclear antibodies evolved in six patients in conjunction with the progression from cirrhosis to hepatoma, and two of three ANA‐positive patients who had the hepatic tumour resected lost ANA from their sera after resection.