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Biomedical subjects

T Morizane

Publications and source records attributed to T Morizane.

At least 55 records · Page 3Linked to original sources
PubMed1987

Transformation of NIH/3T3 cells by DNA from a human hepatoma cell line with integrated hepatitis B virus DNA.

We have studied by means of DNA-mediated gene transfer the transforming activity of the DNA of the human hepatoma cell line HCC-M, which contains genomes of hepatitis B virus (HBV) in integrated form. DNA from HCC-M induced transformed foci on transfection of NIH/3T3 cells. DNAs from primary transformants were capable of inducing secondary transformants. Most of the DNAs of these transformants were demonstrated to contain both human repetitive sequences and HBV DNA, indicating that the transformants had incorporated exogenous human DNA and HBV DNA as well. These results suggest that transformation occurs as the result of the transfer of oncogene which might be closely associated with HBV genome.

Carcinoma, Hepatocellular↗
PubMed1986

New micro-glass-tube leukocyte adherence inhibition assay assessing cell adherence of mononuclear cell subpopulations defined by monoclonal antibodies.

A new micro-glass-tube leukocyte adherence inhibition (LAI) assay which is appropriate for detecting delayed type hypersensitivity in vitro has been developed for human leukocytes. Enumeration of adherent cells is replaced by a cellular radioimmunoassay determining antibody binding of the monoclonal reagents, OKT4, OKT8 and OKM1, to glass-adherent cells, fixed by glutaraldehyde or formaldehyde. An LAI reactivity to purified protein derivative of tuberculin (PPD) was detectable in donors giving a positive PPD skin test with OKT4 reagent, but not with the other two reagents.

Antibodies, Monoclonal↗
PubMed1984

Repeated micro-monocyte adherence inhibition assay: a new technique of leukocyte adherence inhibition to detect tumor immunity in patients with hepatoma.

A microplate leukocyte adherence inhibition (micro-LAI) assay was performed with peripheral blood mononuclear cells obtained from patients with hepatoma and control subjects (including healthy donors and patients with other diseases). Cell extracts of human hepatoma cells (HCC-M) and human hepatic cells (Chang liver cell) in tissue culture were prepared by sonication followed by centrifugation. The supernatants of these two cell lines were used as a specific antigen and a nonspecific antigen, respectively. It was found that monocytes were major indicator cells and that monocytes produced an LAI reaction in the absence of lymphocytes. Therefore, a repeated microplate monocyte adherence inhibition (MAI) assay was developed, in which the monocyte population of adherent cells is increased by removing nonadherent cells after an initial assay in fetal calf serum-containing medium without test antigens, and monocytes are counted selectively as peroxidase-positive cells in a subsequent second assay with test antigens. With regard to sensitivity and reproducibility, the repeated micro-MAI assay is superior to a micro-MAI assay in which the initial assay is omitted although monocytes are selectively counted. With this simple and sensitive technique a hepatoma-associated immune response to the extract of HCC-M was detected in 16 out of 22 patients (73%) with hepatoma, whereas the false-positive rate was 7% (3/41) in all control subjects.

Carcinoma, Hepatocellular↗
PubMed1984

Detection of antibodies to Chang liver cell in sera from patients with chronic liver diseases by 125I-labelled protein A binding assay and the effect of prednisolone and 6-mercaptopurine treatment on the level of the antibodies.

Antibody binding to living Chang liver cell was measured in sera from 71 patients with various chronic liver diseases using 125I-labelled protein A binding assay. The level of antibody binding to Chang liver cell was significantly elevated in sera from patients with chronic active hepatitis (CAH), chronic persistent hepatitis (CPH) and liver cirrhosis as compared to those from healthy donors, but not in sera from patients with fatty liver. There was no detectable antibody binding to HeLa cells in those sera. The antibody binding to Chang liver cell was blocked by a human liver specific protein (LSP) preparation. The levels of antibody binding to Chang liver cell were significantly higher in patients with CAH than patients with CPH. On the other hand, the level of antibody binding to Chang liver cell was significantly decreased in sera from patients with CAH after a treatment with prednisolone (PSL) for 2 months and a subsequent combined administration of 6MP and a maintenance dose of PSL for 1 month. These results suggest that antibodies to Chang liver cell are closely correlated with the activity of chronic liver disease and that PSL and 6MP treatment can reduce the level of the antibodies.

Antibodies↗
PubMed1983

Decreased natural killer activity in patients with liver cirrhosis.

Natural killer (NK) activity of peripheral blood mononuclear cells from patients with non-alcoholic liver cirrhosis (LC) (30 cases), patients with other, non-malignant diseases (41 cases), and healthy subjects (36 cases) were investigated using 51Cr-labelled CCRF-CEM and K562 target cells. NK activity in patients with LC was lower than that in healthy subjects and that in patients with other, non-malignant diseases. The differences were statistically significant by Student's t-test and the Wilcoxon rank test. There was no difference of NK activity between serologically hepatitis B surface antigen (HBsAg)-positive and -negative patients.

Adult↗
PubMed1983

Establishment of a cell line (HCC-M) from a human hepatocellular carcinoma.

A continuous human cell line was established from a hepatocellular carcinoma of an HBsAg-positive Japanese male. The cell line, designated HCC-M, grows as an adhering monolayer with a doubling time of 24 h in medium RPMI-1640 supplemented with 10% FCS and grows with 30% clonal efficiency in soft agar. The cells have been shown by light and electron microscopy to be of epithelial type. When they were transplanted subcutaneously into the back of athymic nude mice (BALB/c, nu/nu), tumors developed at the sites of inoculation, which were shown to be hepatocellular carcinoma, similar in morphology to the original tumor from which they were derived. HCC-M had a chromosome mode of 63 with five identifiable marker chromosomes. HCC-M produced albumin at the 10th passage but this property was lost by the 30th passage. This cell line has not secreted alpha-fetoprotein. Hepatitis B viral particles or HBsAg have not been demonstrated in the cells from the primary culture nor in several subsequent subcultures tested.

Adult↗
PubMed1983

A leukocyte adherence inhibition (LAI) assay of anti-tumor immunity in rats using selective radioimmunological assessment of adherence of T lymphocytes and monocytes.

A new, highly sensitive micro-glass-tube leukocyte adherence inhibition (LAI) assay has been developed to detect anti-tumor sensitization in a rat colon carcinoma system. This technique requires fewer cells and smaller amounts of antigen preparations than the previous glass tube method. The microscopic enumeration of adherent cells is replaced by a cellular radioimmunoassay (CRIA) which utilizes antibodies binding to mononuclear cells (MNC) and 125I-labelled protein A. Antigen preparations were shown to be adsorbed to glass. Precoating of glass vials with fetal calf serum or antigen preparations was shown to cause a major increase in nonspecific LAI. The new LAI technique is designed to minimize such nonspecific LAI. By applying an anti-T-cell monoclonal antibody (McAb), an anti-Ig antiserum and an anti-monocyte (MC) antiserum it became possible to assess selectively the change of adherence of T cells, B cells and monocytes. The specificity of the assay was demonstrated with anti-T-cell and anti-MC reagents in criss-cross experiments using two defined antigens and different tissue extracts as test antigens. The LAI response of peripheral blood MNC was examined after the subcutaneous isografting of 1 X 10(7) X-irradiated DMH-W49 colon carcinoma cells. Six days after sensitization all the rats showed an LAI response detectable with anti-T-cell and anti-MC reagents, which gave considerably higher LAI indices than anti-MNC antiserum. After the subcutaneous isografting of 1 X 10(5) viable DMH-W49 cells, 12 of 24 rats (50%) showed an LAI response detectable with the anti-MNC antiserum during a 3-week follow-up period. On the other hand, an LAI response was detected in all the 14 tumor-inoculated rats tested when the adherence of T lymphocytes and monocytes was assessed selectively.

Adenocarcinoma↗
PubMed1983

Detection of different states of immunity to colon cancer in rats by a leukocyte adherence inhibition (LAI) assay assessing the adherence of T lymphocytes and monocytes selectively.

Peripheral blood mononuclear cells (MNC) of rats, which had received a subcutaneous inoculation of either viable or X-irradiated syngeneic DMH-W49 colon carcinoma cells, were investigated sequentially for sensitization against tumor-associated antigens by a new micro-glass-tube leukocyte adherence inhibition (LAI) assay. The number of adherent cells of MNC subpopulations was estimated by a cellular radioimmunoassay (CRIA), which utilizes anti-monocyte (MC) antiserum or anti-T-cell monoclonal antibody (McAb) and 125I-labelled protein A. LAI reactivity was demonstrated in rats sensitized with X-irradiated tumor cells when assessing the adherence of both T lymphocytes and monocytes. It was found that reactivity with the two different types of indicator cells did not usually coincide in time. The same phenomenon was also observed in tumor-bearing rats although with lower levels of LAI reactivity. In both groups the LAI response detected by anti-T-cell reagent mostly appeared earlier after tumor-cell inoculation than the response demonstrated by anti-MC serum. The T-lymphocyte-associated reactivity also disappeared more rapidly. These results suggest that different states of immune reactivity are reflected in LAI responses involving T lymphocytes and monocytes as indicator cells.

Adenocarcinoma↗
PubMed1980

Impaired T cell function and decreased natural killer activity in patients with liver cirrhosis and their significance in the development of hepatocellular carcinoma.

It is well known that the incidence of hepatocellular carcinoma (HCC) in patients with liver cirrhosis (LC) is very high. We investigated the immunological state in patients with LC and HCC. T cell population of peripheral blood lymphocytes (PBL) and blast transformation of PBL by phytohaemagglutinin (PHA) were significantly decreased in patients with LC. Natural killer activity against HeLa cell was also significantly decreased in these patients. These results suggest that immunological surveillance is impaired in patients with LC and this may be one of the aetiological factors in genesis of hepatocellular carcinoma in patients with LC.

Carcinoma, Hepatocellular↗
PubMed1980

Specific immunodiagnosis of hepatoma by tube leukocyte adherence inhibition assay and a modified method of repeated tube leukocyte adherence inhibition assay.

Tube leukocyte adherence inhibition (LAI) assays were performed with normal liver extract as nonspecific antigen and with hepatoma extract as specific antigen in patients with hepatoma. Titration experiments revealed that the optimal extract concentration was 400 micrograms/ml when expressing the results in terms of a nonadherence index. The results of tube LAI assays were positive in 26 of 40 cases (65%) of hepatoma. The results were negative in all cases of other liver diseases and other cancers. The tube LAI assay was repeated after discarding the non-adherent cells in the initial tube LAI assay with normal liver extract. The nonadherence index of the repeated tube LAI assay we devised was significantly higher than that of the original tube LAI assay (p less than 0.001) in patients with hepatoma. Ten of 12 patients with hepatoma in whom the results of the original tube LAI assay were negative showed positive results in the repeated tube LAI assay. The present study suggests that the problem of false-negative results in tube LAI assay can be solved by repeating the tube LAI assay.

Antigens, Neoplasm↗
PubMed1978

Leukocyte migration inhibition induced by the combination of drug and a liver constituent in patients with drug-induced hepatitis.

The leukocyte migration inhibition test in agarose medium was performed in 23 cases of clinically diagnosed drug-induced hepatitis. When the test antigen was the combination of soluble phase of a liver homogenate fractionated by Sephadex G-100 which should have contained liver specific antigen and the offending drug the leukocyte migration was inhibited in 86% of cases. Whereas none of 12 cases of drug allergy without hepatic injury showed a positive result with the same combination of antigens. Other organ homogenate-muscle and kidney-never gave positive results when mixed with the offending drugs in cases of drug-induced hepatitis. It was concluded that in hypersensitivity type drug-induced hepatitis cell-mediated immunity might be established to the complex of liver specific antigen and the drug.

Adolescent↗