Takashi Tsukishiro

Effect of branched‐chain amino acids on the composition and cytolytic activity of liver‐associated lymphocytes in rats

Background : Although branched‐chain aminoacids (BCAA) are reported to be effective in prolongation of the mean survival time of patients with liver cirrhosis, it is not clear whether BCAA could influence the immune function in those patients.

Evaluation of transcatheter arterial embolization with epirubicin-lipiodol emulsion for hepatocellular carcinoma*

SummaryA total of 18 patients with hepatocellular carcinoma (HCC) were treated by transcatheter arterial embolization (TAE) with a 4′-epi-doxorubicin (EDX)-lipiodol emulsion. Infusion of the EDX-lipiodol emulsion (EDX-L) via the hepatic artery was followed by the injection of gelatin sponge in 12 cases. The response and survival of these 12 patients following EDX-L treatment were compared with those of 42 subjects treated with a doxorubicinlipiodol emulsion (DX-L) and those of 23 patients treated by TAE with gelatin sponge (GS) only. In the group treated with EDX-L, nine cases were AFP-positive in sera and four showed a decrease in serum AFP values to less than 10% of the pretreatment level. Seven cases showed a partial response, and nine cases showed no change in the size of the tumor. In the group treated with EDX-L, nine cases are alive, and the oldest has survived for more than 431 days since the treatment. The half-year survival value was 57%, and the 1-year survival value was 49%. These values did not differ significantly from those calculated for the group treated with DX-L. The 1-year survival value determined for patients treated with a lipiodol emulsion (EDX-L or DX-L) followed by GS was 65%, and the 2-year survival value was 39%. These results rates are significantly better than those obtained in patients treated with GS only (1-year survival, 39%; 2-year survival, 13%).

Analyses of proliferating cell nuclear antigen‐positive cells in hepatocellular carcinoma: Comparisons with clinical findings

Abstract Proliferating tumour cells in 92 patients with hepatocellular carcinoma (HCC) were identified by an immunohistochemical method using a monoclonal antibody against proliferating cell nuclear antigen (PCNA). The rate of PCNA‐positive cells in HCC tissues was positively correlated with histological grade and the tumour size and T factor of the tumour.

Modulation by cytokines on the surface expression and shedding of intercellular adhesion molecule-1 (ICAM-1) in human liver cancer cell lines

Abstract Surface expression and shedding of intercellular adhesion molecule-1 (ICAM-1) in the absence and presence of cytokines were measured in seven human liver cancer cell lines. All cancer cell lines were positive for ICAM-1 on the surface of the cells, and the percentages of positive cells ranged from 60 to 97% as analyzed by flow cytometry. Mean fluorescence was increased in the presence of IL-1β, TNF-α, IFN-α or IFN-γ, and the effect was augmented in a dose-dependent fashion. In the culture medium of all cancer cell lines, a soluble form of ICAM-1 was detected even in the absence of cytokines by enzyme-linked immunosorbent assay, and the concentration was increased by IL-1β, TNF-α, or IFN-γ. In two of seven cancer cell lines, the patterns of modulation by cytokines on the surface expression of ICAM-1 and the shedding of the molecules were clearly distinct, suggesting that the mechanisms for these two phenomena might be different. Soluble ICAM-1 was recently reported to suppress non-MHC-restricted cytotoxicity mediated by natural killer and lymphokine-activated killer cells. Our present data suggest that human liver cancer cells may escape from host immune surveillance by shedding of ICAM-1.

Variable Region Usage in T Lymphocytes Infiltrating Liver Tissues of Chronic Liver Diseases

To analyze the T cell repertoire of liver-infiltrating lymphocytes, 35 liver biopsy specimens taken for diagnostic purposes were stained by immunohistochemical technique using monoclonal antibodies (mAb) against CD3, CD8, CD45RO, and CD45RA and seven different variable (V) regions of the T cell receptor (TCR). Major populations of the infiltrating lymphocytes were CD3- and CD45RO- positive cells. The mean percentage (±SD) of Vβ5.1-positive cells was significantly (P < 0.05) higher in chronic hepatitis C (18.2 ± 13.3%, n = 15) than that in either chronic hepatitis B (5.7 ± 6.0%, n = 12) or autoimmune hepatitis (1.3 ± 1.2%, n = 3). In cases with primary biliary cirrhosis, only one case showed a high percentage (66%) of Vβ5.1-positive cells with mean ± SD of 19.3 ± 26.2% (n = 5). The percentages of Vβ5.1-positive cells correlated inversely with the Knodell histological activity index scores in cases with chronic hepatitis C. These data suggest that Vβ5.1-positive T cells may play a regulatory role in the immuno-pathogenesis of chronic hepatitis C.

Serum interleukin-8 levels in patients with hepatocellular carcinoma

Abstract Serum levels of interleukin-8 (IL-8) in 73 patients with hepatocellular carcinoma (HCC), 24 with liver cirrhosis (LC) and 18 with chronic hepatitis (CH), and in 20 healthy controls were measured by an enzyme-linked immunosorbent assay. The mean (± SD) level of serum IL-8 in patients with HCC was 48.4 ± 60.8 pg/ml, which were significantly (P

Induction of allogeneic tumour‐ and lymphokine‐activated lymphocytes against hepatocellular carcinoma

For clinical application of adoptive immunotherapy against hepatocellular carcinoma (HCC), it is not easy to prepare tumour specific effector cells such as cytotoxic T lymphocytes (CTL). To induce potent and broad‐spectrum effectors, allogeneic cultured hepatoma cell lines (JHH‐4 and HuH‐6) were used as stimulators of peripheral blood lymphocytes (PBL) instead of autologous HCC cells. Allogeneic tumour‐ and lymphokine‐activated killer cells (ATLAK) were generated by a mixed culture of lymphocytes and allogeneic cultured tumour cells with recombinant interleukin‐2 (rIL‐2). The tumour‐killing activity of ATLAK induced by HuH‐6 was confirmed against HuH‐6 and other different HCC cell lines (JHH‐2, HuH‐7 and PLC). These activated lymphocytes were significantly more potent than lymphokine‐activated killer cells (LAK) in [51Cr]‐releasing assay. The JHH‐4 stimulated ATLAK was reactive not only with JHH‐4 but also with JHH‐2. The lysis of allogeneic targets could be partially inhibited by anti‐CD8 and anti‐CD3 but not by anti‐CD4. Anti‐tumour cytotoxicity in these cultures might be mediated by CD3+CD56‐ and CD3+CD56+ effectors. These results imply that adoptive immunotherapy for HCC with ATLAK may be more feasible than that with LAK.

Hepatocellular carcinoma in 13 patients with hepatitis C virus-associated chronic hepatitis

Abstract Thirteen of 81 patients with chronic hepatitis and positive hepatitis C virus (HCV) antibody developed hepatocellular carcinoma (HCC) during a follow‐up period of 54 ± 38 months. The histopathological findings in HCC‐bearing liver in these patients included six cases of chronic persistent hepatitis [CPH; mean hepatitis activity index (HAI) score: 5.8] and seven cases of chronic aggressive hepatitis (CAH) 2A, or 2B (HAI) score: 13.6). Multiple biopsies of the liver in six cases revealed that five cases, including four with CPH at the time of HCC diagnosis, previously had histopathological findings identical to CAH 2A, and another case constantly had CPH during the 8‐year follow‐up. These findings suggest that HCV‐associated HCC can occur even in patients with HCV antibody positivity and inactive or mild chronic hepatitis. This is of interest in the pathogenetic mechanisms of HCV‐associated HCC.

Cytokine effect on the concentrations of interleukin-8 (IL-8) in the culture medium of human liver tumor cell lines

Abstract The concentrations of interleukin-8 (IL-8) in the culture medium of seven human liver tumor cell lines in the absence or presence of cytokines were measured by enzyme-linked immunosorbent assay. IL-8 was detected in all cultures even in the absence of cytokines, and the concentrations ranged from 270 pg/ml to 62 ng/ml. The IL-8 production was stimulated by the addition of IL-1β or TNF-α into the cultures of all cell lines, while IFN-α and IFN-γ suppressed the production in some of the tumor cell lines. IL-8, which was originally described as a chemotactic factor for neutrophils and T lymphocytes, was recently reported to have a strong angiogenic potential. Therefore, IL-8 production by liver tumor cells is thought to be necessary for their vascularization. Inhibition of IL-8 production by cytokines may have therapeutic potential by suppressing neo-vascularization.

A Case of Cytomegalovirus‐Induced Gastric Lesions

Abstract: A 26‐year‐old woman was admitted to our hospital with epigastric pain, a husky voice, high‐grade fever, hepatosplenomegaly and enlargement of the kidneys. An upper gastrointestinal endoscopy revealed scattered non‐circular erythematous erosions, up to 5mm in diameter, in the gastric body and fornix. A histological examination of the biopsied specimens stained with hematoxylin and eosin showed the presence of intranuclear inclusion bodies in the glandular epithelial cells. Cytomegalovirus (CMV) antigen was found in the inclusion bodies using the enzyme‐labeled antibody method. Subsequently, a needle biopsy taken from the kidney revealed a non‐Hodgkins lymphoma. Our review of the literature indicates that CMV‐induced gastric lesions are endoscopically characteristic and can be diagnosed from their multiple, erythematous, non‐circular depressed lesions.