Koji Yabu

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.

Type C chronic hepatitis associated with thrombocytopenia in two patients

Abstract Portal hypertension in the presence of chronic hepatitis is generally thought to develop during the progression of the chronic hepatitis to cirrhosis. Before the establishment of assays for diagnosing hepatitis C virus infection, such a case of portal hypertension without liver cirrhosis could be misdiagnosed as idiopathic portal hypertension. It had not fully determined whether portal hypertension might precede the onset of cirrhosis in type C chronic hepatitis. This report presents two cases of women with chronic hepatitis C who developed severe thrombocytopenia; each showed splenomegaly and hypersplenism due to portal hypertension. Angiographic study and histological analysis were conducted to determine the cause of the portal hypertension. Histological evaluation showed an intrahepatic presinusoidal block pattern and fibrotic changes in the periportal area, but no evidence of liver cirrhosis or of other incidental complications such as idiopathic portal hypertension. Both of these patients exhibited normal platelet counts after splenectomy. Thus, type C chronic hepatitis can lead to portal hypertension, as demonstrated in these two patients.

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.

Caroli's disease in three siblings

SummaryThree sisters with cystic dilatation of the intrahepatic bile ducts (Caroli’s disease) are reported. The index case, a 41-year-old woman with remittent high fever and right upper quadrant abdominal pain, was diagnosed as Caroli’s disease with hepatic lithiasis and cholangitis based on findings of ultrasonography, computed tomography and endoscopie retrograde cholangiography. Her two older sisters were also examined and found to have the same disease without clinical symptoms. Their symptoms, locations of the dilated ducts and complications all varied. The hereditary mode of Caroli’s disease in 13 families (32 cases) reported in the world literature including our study was examined. While Caroli’s disease is thought to be an autosomal recessive disease, a conclusion on the hereditary mode of transmission could not be made in this study because of an insufficient investigation of family members, especially the parents.

Cellular FITC-linked immunospecific assay (Cell-FLISA) for detection of monoclonal antibodies against cell-surface antigens

A cellular fluorescein isothiocyanate (FITC)-linked immunospecific assay (Cell-FLISA) has been established using the recently developed fluorophotometer for microplates. In the Cell-FLISA system, monoclonal antibodies specific for the surface antigens of live cells are detected by measuring the fluorescence intensity of an FITC-labeled second antibody: goat anti-mouse immunoglobulin antibody. It takes only 2 min to count 96 samples in microplate wells using the fluorophotometer for microplates. Moreover, by this system, the analysis is finished within 2 hr. Thus, the Cell-FLISA system has advantages in screening a large number of samples, such as hybridoma cell lines secreting monoclonal antibodies against cell-surface antigens.

Immunological responses against an autologous human hepatocellular carcinoma cell line

We performed a detailed analysis of immune responses in a hepatocellular carcinoma (HCC) cell line and effector cells obtained from a patient with HCC. We examined the cytotoxic activity of natural killer (NK) cells, lymphokine‐activated killer (LAK) cells and cytotoxic T lymphocytes (CTL) against an autologous tumour cell line (SUHC‐1) to investigate the immune mechanism of human lymphocytes against HCC cells. Cytotoxic T lymphocytes were induced by co‐culturing of peripheral blood lymphocytes (PBL) and SUHC‐1 cells, mixed lymphocyte and tumour cell culture (MLTC). The susceptibility of SUHC‐1 to NK and LAK cells was similar to that of other allogeneic cell lines, such as K562, PLC/PRF/5 and Mahlavu. Effector cells induced in the primary MLTC had high cytotoxic acitivity but were not specific for SUHC‐1. Cytotoxic T lymphocytes with specific activity against SUHC‐1 were induced after PBL were stimulated five times at 7–10 day intervals with SUHC‐1 and low‐dose recombinant interleukin‐2 (rIL‐2), suggesting that as the culture progressed, broadly reactive effector cells disappeared and specific effector cells survived. The specific effector cells were identified as CD3+/CD4+ and CD+/CD8+ T‐lymphocyte subsets. The recognition mechanisms of CD3+/CD4+ CTL remain unresolved because the cytotoxicities were not inhibited by anti‐CD4 and anti‐major histocompatibility complex (MHC) class II monoclonal antibodies (MoAb). Treatment of cells with anti‐CD3, anti‐CD8 and anti‐MHC class I MoAb partially inhibited lysis. These results demonstrated that the T‐cell receptor (TCR)/CD3 complex appeared to be involved in SUHC‐1 specific antigen recognition and antigen recognition of CD3+/CD8+ CTL was MHC class I restricted.

Lymphocyte proliferative responses to recombinant hepatitis C virus antigens in patients with chronic hepatitis C

The purpose of the present study was to analyse lymphocyte proliferative responses to recombinant hepatitis C virus (HCV) antigens in chronic hepatitis C. Four recombinant peptides derived from the NS3, core, E1 and E2/NS1 regions of the HCV genome were used as antigens in lymphocyte proliferative responses. Forty‐two patients, classified into various sub‐groups, and 17 healthy control subjects were tested and the specific response was expressed as a stimulation index. Responses were analysed with alanine aminotransferase (ALT) level and histological diagnosis. NS3‐ and core‐antigen specific responses in all patient groups were significantly higher than in the healthy control group. E1‐ and E2/NS1‐antigen‐specific responses in the patient group with ALT levels exceeding 100 IU/L were significantly higher than those in other patient groups. Histological diagnosis was not correlated to the intensity of the core‐ and NS3‐specific responses. E1‐ and E2/NS1‐antigens induced significantly elevated responses in patients with chronic active hepatitis and liver cirrhosis compared with results in the healthy control group and in patients with chronic persistent hepatitis. In conclusion, the significantly elevated responses to core‐ and NS3‐antigens may be related to HCV infection and such responses to E1‐ and E2/NS1‐antigens could be related to the severity and activity of the disease.

A case of asymptomatic primary biliary cirrhosis with an initial presenting feature of localized gastric varices.

SummaryA case of asymptomatic primary biliary cirrhosis (PBC) with an initial presenting feature of localized gastric varices is reported.The patient, 64 years old female, underwent a barium meal examination because of ill-defined abdominal complaints and was found to have gastric varices localized at the cardia but no esophageal varices. Her blood chemistry showed high values of biliary tract enzymes such as alkaline phosphatase and gamma-glutamyl transpeptidase, but the serum bilirubin level was almost normal. Serum anti-mitochondrial antibody was positive. Histological findings of the surgically biopsied liver specimen were compatible with PBC. The clinical implication of gastric varices in PBC is discussed.

In Vitro Immune Responses Specific for NS3 Regional Peptide (C7) and Core Regional Peptide (C11) of HCV in Chronic Hepatitis C

Peripheral blood lymphocyte (PBL)-proliferative responses against NS3 regional peptide (C7) and core regional peptide (C11) of hepatitis C virus were analyzed in chronic hepatitis C. The PBLs of the patients with elevated alanine aminotransferase (ALT) levels (ALT > 45U/L) responded signficantly against C11 antigen, while PBLs of all patients responded against C7 irrespective of the ALT level. Histological differences among the patients did not affect the proliferative responses against both antigens to a statistically significant degree. The production of anti-C7 and anti-C11 antibodies in vitro was also examined. These antibodies, produced by the PBLs, were detected in one-third of the patients, those with lower ALT levels and relatively weaker proliferative responses. Thus, antibody production in vitro did not correlate to the proliferative responses. These discrepancies may provide a clue to remission in chronic hepatitis C.

Studies on lymphocyte subsets for the differential diagnosis between non-A, non-B viral hepatitis and drug induced liver injury, and on the role of Ia positive cell in lymphocyte stimulation test.

薬剤性肝障害の診断上,ウイルスマーカーの確立されていない非A非B型ウイルス肝炎との鑑別が問題となる.そこで,薬剤性肝障害と非A非B型ウイルス肝炎の末梢血リンパ球亜分画を測定し,その鑑別診断的意義について検討するとともに薬剤性肝障害例におけるリンパ球刺激試験(LST)の免疫学的機序を解析した.薬剤性肝障害では,急性期OKT8陽性細胞の増加によりT4/T8比が低下し,回復期正常化している.それに対し,非A非B型ウイルス肝炎では急性期より正常範囲であった.このことから,T4/T8比の測定により両者の鑑別がある程度可能であると考えられた.また,LSTにおいてIa抗原陽性細胞が抗原呈示細胞として,薬剤特異的T細胞の増殖反応に重要な役割を演じていることが示された.