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Biomedical subjects

J Minowada

Publications and source records attributed to J Minowada.

At least 109 records · Page 6Linked to original sources

Expression of the human T-cell-specific tyrosine kinase YT16 (lck) message in leukemic T-cell lines.

To evaluate the role of the human T-cell-specific tyrosine kinase lck (YT16), we measured the levels of lck expression in thymocytes, peripheral T-cells, and leukemia T-cell lines which are arrested at different stages of thymic differentiation. The results indicate that higher levels of the lck message can be found in total thymocytes and T-cells arrested at Stage III (thymocytes that have rearranged their alpha, beta, and gamma chain T-cell receptor genes). A 20-fold lower level of these transcripts, however, can be found in cells derived from Stage I cells (thymocytes with germline alpha, beta, and gamma genes), 4 times less messages in Stage II cells (thymocytes with rearranged gamma and beta chain genes, but with germline alpha chain genes), and a 4-fold lower amount of RNA in Stage IV cells (mature lymphocytes). Culture of these cells with a variety of inducing reagents indicated that leukemia cell lines of only Stages I and II can be induced to increase their levels of YT16 expression by the addition of tetradecanoyl phorbol-13-acetate. These results suggest that there may be a developmental regulation of these tyrosine kinase messages during T-cell ontogeny. The high level of these transcripts in more mature T-cell leukemia lines suggests that they may primarily play a role in the proliferative controls of these cells.

Antibodies, Monoclonal↗

Enhancement of human immunodeficiency virus production by natural lymphotoxin.

The effect of natural lymphotoxin (n-LT) on CD4+ MOLT-4 cells and those cells producing human immunodeficiency virus (HIV), MOLT-4/HIVHTLV-IIIB cells, was studied. Four days after treatment with n-LT, a significant cytotoxic/cytostatic effect was observed predominantly on MOLT-4/HIVHTLV-IIIB cells. Furthermore, with regard to the production of HIV, an almost 3- to 5-fold increase of viral particles was observed in MOLT-4/HIVHTLV-IIIB cells 6 h after treatment with n-LT. These data indicate the possibility that this cytotoxic factor is one of the responsible molecules in the pathogenesis of the acquired immunodeficiency syndrome (AIDS).

Antibodies, Monoclonal↗

Routine immunophenotyping of acute leukaemias.

Recent progress in immunophenotyping includes the availability of monoclonal antibodies (MAbs), knowledge of specificity and reactivity patterns of these reagents, and the technical improvements and standardization of immunofluorescence and immunocytology staining procedures, including flow cytometry. These advances have contributed significantly to the establishment of immunophenotyping as an essential diagnostic tool in the differential diagnosis of types of acute leukaemia. Immunophenotyping allows for the objective and reproducible distinction of acute lymphoblastic leukaemia (ALL) from acute myeloblastic leukaemia (AML) and of T-lineage from B-lineage ALL. Immunologically defined ALL and AML subtypes have been found to convey prognostic significance. Using cell lineage-specific and differentiation stage-specific MAbs, cases of T- and B-lineage ALL and of AML can be further classified into a number of different subtypes. Routine immunophenotyping concentrates on the diagnostic enquiry into a few major, clinically relevant subtypes; only a limited number of crucial reagents are employed that are commercially available. The simplification and standardization of discriminatory immunomarker panels make immunophenotyping a reliable diagnostic instrument for the provision of critical data to make a differential diagnosis. An effort to identify the nature and origin of the blast cells precisely, immunological typing definitely plays an important part in the multiple-marker analysis of acute leukaemia (morphology, cytochemistry, karyotyping, genotyping) for applied diagnostic and fundamental research purposes.

Acute Disease↗

Quantitative and qualitative studies of leukaemic cell dipeptidylpeptidases II and IV.

Cellular concentrations of dipeptidylpeptidases (DAP) II and IV were determined by fluorimetric assay in 152 cases of leukaemia and 24 permanent T-cell lines. Whilst its estimation appears to have few diagnostic applications, it was found that DAPII levels were generally higher in prolymphocytoid CLL variants (CLL-Pro) compared with "typical" CLL cases. Qualitatively, DAPII extracted from leukaemic B- and T-cells did not differ significantly with respect to molecular weight (Mr 82 kD: FPLC gel filtration) or substrate affinity (Km). Similarly, a single molecular weight species (Mr 189 kD) of DAPIV was also detected in all leukaemic sonicates examined although, in quantitative terms, increased DAPIV levels were primarily associated with a subgroup of CD4-positive postthymic malignancies. Studies of immature T-cell proliferations however indicated that DAPIV expression was unrelated to either stage of immunological differentiation or CD4 expression. No evidence of lineage restriction was found for either enzyme and it is concluded that variations in DAP cytochemical staining patterns reflect quantitative rather than qualitative differences.

Antigens, Differentiation, T-Lymphocyte↗

Ki-ras-2 in acute lymphoblastic leukemia cells with chromosome change at 12p12.

Cytogenetic and molecular investigations were performed on a pre-B-lymphoblastic acute leukemic cell line (NALM-6). The NALM-6 cells contained a del(5)(q32) and an ins(12)(p12;?), chromosomal material of unknown origin being inserted between subbands 12p12.1 and 12p12.2. Chromosomal in situ hybridization using a 3.0-kb c-Ki-ras-2 probe showed a significant accumulation of grains on the proximal portion of the inserted chromosomal material (12p12.1), as well as on the normal chromosome #12 with a peak at 12p11p12. The signal intensity obtained after hybridization of the c-Ki-ras-2 specific probe to the NALM-6 cells DNA is comparable with the intensity of the signal after hybridization of the same probe with the control cell line (MC/B) DNA. The findings indicate that the c-Ki-ras-2 gene is neither amplified nor transposed in the NALM-6 cells.

Chromosome Banding↗

Human leukemia cell lines--clinical and theoretical significances.

Establishment and total characterization of permanent growth-factor independent human leukemia cell lines are reviewed. Among others, a few significant contributions in the utility of leukemia cell lines described include establishment of immunodiagnosis of human leukemias and lymphomas, discovery of EB virus and of human retroviruses, proposed model of normal hematopoietic cell differentiation scheme, lymphokine-monokine production, identification and characterization, studies on genes responsible for immunobiological and growth regulatory molecules, and pharmacotoxicological and kinetic research. It is conceivable that the future leads by the utility of human leukemia cell lines to the advancement of research are indeed unlimited.

Cell Differentiation↗

Quantitative and qualitative enzyme studies of Hodgkin's disease-derived cell lines.

Any extensive analysis of Hodgkin (H) and Reed-Sternberg (RS) cells is limited by the scarcity of available material and by the common contamination with "by-stander" cells. The establishment of Hodgkin's disease-derived cell lines provides the opportunity to undertake studies in which large numbers of cells are required, as these cell lines are by definition monoclonal populations with unlimited cell growth. In this study, we analyzed the enzyme profiles of eight Hodgkin's disease cell lines (Co, Ho, Fox, HDLM-2, KM-H2, L428, L540, and L591) whereby cellular alpha-naphthyl acetate esterases, acid phosphatases, and dipeptidylpeptidase IV were examined quantitatively or qualitatively by IEF or chromatographic techniques. The results indicate that all of the H-RS cell lines examined had enzymatic features typical for lymphoid cells and, in particular, a monocyte/histiocyte origin of the cell lines could be excluded. Extrapolation of the available immunological, molecular biological, and enzymological evidence gained in vitro on cultured representatives of H-RS cells suggests a lymphoid origin for in vivo H-RS cells.

Acid Phosphatase↗

Genotypes and immunophenotypes of Hodgkin's disease-derived cell lines.

This report describes the geno- and immunophenotypic analysis of the Hodgkin's disease-derived cell lines HDLM-2, KM-H2, and L-428. The lines were all positive for the antigens CD15 (Leu-M1), CD30 (Ki-1), Hefi-1 (antigen detected by a monoclonal antibody produced against L-428), HLA class I and II, and activation/proliferation markers. The cells from all 3 cell lines lacked almost all cell lineage-associated/specific markers: HDLM-2 was only CD2+, KM-H2 was only CD9+ and CD21+, and L-428 was negative for all the specific markers tested. Genomic analysis of HDLM-2 cells revealed monoclonal rearrangements of T cell receptor beta and gamma loci and germ line configuration of immunoglobulin genes. Immunoglobulin heavy chain genes were rearranged in KM-H2 and L-428. These data suggest a possible lymphoid origin for HDLM-2, KM-H2, and L-428. Although the data presented do not provide formal proof of a lymphoid nature of Hodgkin and Reed-Sternberg cells and do not unequivocally exclude a derivation from other hematopoietic cells, extrapolation of the results from the in vitro cultures to the in vivo situation suggests a lymphoid (T or B cell) origin of these cells.

Antigens, Differentiation, T-Lymphocyte↗

Loss of a Hu-ets-1 allele in human leukemia cell lines ML-1, -2, and -3 with a chromosome change at 11q24.

The ML cell lines (ML-1, -2, and -3) were derived from the cells of a patient with T-cell malignant lymphoma who developed acute myeloblastic leukemia and whose cells showed a primary chromosome change at band 11q24. Surface marker studies of the ML cells showed that they had both myeloid (MCS-1, MCS-2, and OKM-1) and some T-lymphocyte (3A1/Leu-9 and OKT-4/Leu-3a) characteristics. Molecular studies on these lines were performed in order to determine the possible involvement of the Hu-ets-1 gene, since it is located at band 11q23----q24. All ML cell lines showed half the intensity of the Hu-ets-1 DNA bands as compared to those of controls (karyotypically normal B-cell lines). In contrast, DNAs from leukemia cells with t(4;11)(q21;q23) or t(1;11)(q21;q23 or q24) showed no rearrangement, deletion, or amplification of the ets-1 gene. These findings indicate that a chromosome region (11q24----qter), including the Hu-ets-1 gene, of the ML cells is deleted as a result of the primary cytogenetic change and that heterogeneity is present in the mechanism of human leukemia involving the 11q23----q24 region.

Adult↗

A serine proteinase inhibitor produced by an HTLV I virus-transformed human T lymphocyte line.

A previous report from our laboratory indicated that a proteinase inhibitor is produced by rabbit T lymphocytes. We now report that a human T cell line, C91/PL, produces a proteinase inhibitor which inhibits the enzymatic activity of trypsin and kallikrein. This newly identified proteinase inhibitor (LPI 1) did not inhibit the enzymatic activity of four other serine proteinases (thrombin, plasmin, chymotrypsin, or pancreatic elastase), a thiol proteinase (papain), or a carboxyl proteinase (pepsin). Active synthesis of LPI 1 by the C91/PL cell line was shown by the appearance of similar levels of inhibitory activity in sequential cell supernatants, lack of appearance of inhibitor in supernatants of cells killed by heat or sodium azide or of viable cells in the presence of cyclohexamide, and incorporation of a radiolabeled amino acid into newly synthesized inhibitor. Although both the inhibitor of rabbit origin and of human origin are proteins produced by T cells and have similar inhibitory specificity, important differences were observed: LPI 1 is sensitive to boiling and the two inhibitors migrate differently upon electrophoresis in substrate-containing polyacrylamide gel. Furthermore, LPI 1 was produced by a cell line of the T4 phenotype which had been established by in vitro viral transformation of human cord blood lymphocytes with HTLV 1 whereas the inhibitor of rabbit origin was produced by normal splenic T cells. Three other human T cell lines of the T4 phenotype, MOLT-13, KE-37, and HPB-ALL, from patients with acute lymphoblastic leukemia did not produce a proteinase inhibitor. Thus, the production of proteinase inhibitors does not appear to be a general characteristic of human T cell lines nor of the T4 subset. Proteinase inhibitors produced by T cells may have an immunoregulatory role in proteinase-mediated physiological processes.

Aprotinin↗

Occurrence of particular isoenzymes in fresh and cultured leukemia-lymphoma cells. III. Esterase isoenzyme in monocytes.

The expression of a particular alpha-naphthyl acetate esterase isoenzyme which is specific for monocytes was examined in a panel of cultured leukemia-lymphoma cell lines (n = 88), freshly obtained leukemia-lymphoma cells (n = 527), and in fresh (n = 10) and cultured (n = 22) leukemia cells treated with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). The sodium fluoride-sensitive isoenzyme was separated by isoelectric focusing on horizontal thin-layer polyacrylamide gels. The esterase isoenzyme was not detected in untreated or TPA-treated lymphoid, erythroid, or Hodgkin's disease-derived cell lines, but was seen in leukemia cell lines of monocytic origin. TPA induced the new expression of this marker isoenzyme in two leukemia cell lines of promyelocytic and erythroid origin that are known to differentiate along the monocytic-macrophage cell lineage; TPA stimulation increased the staining intensity of the band in monocytoid cell lines. This esterase isoenzyme was found in 92% of the cases classified morphologically as acute myelomonocytic or monocytic leukemia, but only in 3% of the non-monocytic acute myeloid leukemias. All lymphoid or erythroid leukemias or lymphomas were negative. Treatment with TPA of AML and CML cells, which commonly differentiate to monocyte/macrophage-like cells, showed de novo the monocyte-specific isoenzyme. It is concluded that this isoenzyme is a characteristic marker for monocytic leukemia cells and will be a useful tool for the discriminatory identification of the monocytic element in normal and leukemic cells.

B-Lymphocytes↗

Phenotypic changes associated with chemically induced differentiation of a hairy cell leukemia cell line.

In vitro chemically induced differentiation of the JOK-1 cell line, derived from a patient with hairy cell leukemia, has been studied. Non-induced JOK-1 cells expressed sIgM, B-1, CALLA, Ia, and C3 receptors. Following exposure to 2% DMSO, or 100 ng/ml 12-O-tetradecanoyl-phorbol-13-acetate (TPA), the cells underwent a phenotypic change from lymphoblastoid to monocytoid. In the presence of TPA, they became adherent and developed dendritic processes. Associated with these changes was a decrease in sIgM and CALLA and variable expression of C3 receptors. No monocyte/macrophage-associated antigens were detected. These results suggest that the JOK-1 cell line may have a limited ability to differentiate but is not mature enough to complete the differentiation process.

Antigens, Surface↗

T-leukemia cell lines CCRF-CEM, HPB-ALL, JM and MOLT-4: changes in isoenzyme profiles during induction of differentiation.

Biochemical analysis has been used to monitor the induction of differentiation in cultured human T-leukemia cell lines (CCRF-CEM, HPB-ALL, JM and MOLT-4) by the phorbolester 12-0-tetradecanoylphorbol 13-acetate (TPA). The isoenzymes of carboxylic esterase, acid phosphatase, hexosaminidase and lactate dehydrogenase were separated by isoelectric focusing on horizontal thin-layer polyacrylamide gels and stained by histo-cytochemical methods. TPA inhibited the proliferative activity in all four cell lines and led to aggregation of cells seen as floating clusters. TPA induced an increase in number and staining intensity of isoenzymes of all four enzymes in the cell lines studied. This corresponds to an induced isoenzymatic maturation as the progressive increase in number and staining intensity of the isoenzymes parallels the differentiation along the T-cell pathway. However, regardless of the initial stage of arrested differentiation, the cell lines could be induced only to differentiate to a certain more mature stage, but could not be triggered to differentiate terminally with regard to expression of isoenzyme patterns.

Cell Aggregation↗

Isoenzyme studies in human leukemia-lymphoma cell lines--V. Induction of differentiation by T-cell derived differentiation-inducing activity.

A panel of human leukemia cell lines from various lineages (T-cell, pre B- and Non-T/Non-B cell, myelomonocytic and erythroleukemia cell lines) were utilised as model systems of the distant effects of differentiation-inducing activity (DIA) produced by the T-leukemia cell line HUT-102. DIA inhibited cell proliferation and induced distinct morphological changes which were more pronounced in the myelomonocytic and erythroleukemia cell lines than in the lymphoid cell lines. DIA triggered in the myelomonocytic and erythroleukemia cell lines an increase in the number of NBT-reducing cells and caused strong adherence to plastic surface. The T-cell lines showed aggregation of cells in floating clusters. In the isoenzyme analysis of the enzymes carboxylic esterase and acid phosphatase, it was found that DIA stimulated the new expression of isoenzymes and a stronger staining intensity of several isoenzyme bands in all cell lines, however, at varying degrees. HL-60 and HEL displayed newly a monocyte-specific isoenzyme. Several myelomonocytic and erythroleukemia cell lines were triggered to express the tartrate-resistant acid phosphatase isoenzyme. The cell kinetic, morphological, functional and isoenzymatic data demonstrated that DIA effected the development of the different blood cell types. However, it appears that the cells reached a new differentiation block after acquired expression of differentiation-linked features; the lymphoid cell lines were more limited in their response to DIA than the myeloid and erythroid cells.

Acid Phosphatase↗

Human-fms gene is retained in acute lymphoblastic leukemia cells with del(5)(q32).

Cytogenetic and molecular investigations of NALM 6 cells (a pre-B-lymphoblastic acute leukemia cell line) revealed them to contain both alleles of the c-fms gene, though the cells had chromosomal changes of 5q- and 12p+. The amount of DNA fragments hybridized to the 1.4 kb PstI/PstI v-fms probe in the NALM 6 cells was approximately the same, when compared with cells of an Epstein-Barr virus-transformed lymphoblastoid cell line with a normal karyotype. Chromosome banding analysis revealed that the breakpoint of the 5q- in the NALM 6 cells was at the proximal portion of the 5q32 band. Chromosomal in situ hybridization of NALM 6 cells showed a significant accumulation of grains on the terminal portions of the abnormal 5q- chromosomes (5q32), as well as on the normal chromosomes #5 with a peak at 5q32-q33. These findings indicate that the human c-fms gene is not deleted in the lymphoblastic leukemia cells with a 5q- studied by us and that it does not show rearrangement or amplification. Thus, the results indicate that a difference in the dosage of the c-fms gene in acute lymphoblastic leukemia cells with the 5q- versus that in cells with the 5q- change in nonlymphocytic neoplasia; in the latter a hemizgosity of the c-fms gene has been suggested.

Cell Line↗

Expression of FMC7 antigen and tartrate-resistant acid phosphatase isoenzyme in cases of B-lymphoproliferative diseases.

A panel of different B-cell malignancies representing various stages of B-cell differentiation were analyzed for the expression of an antigen labeled by the monoclonal antibody FMC7 and of tartrate-resistant acid phosphatase (TracP) activity. The FMC7 antigen and TracP were not found on early immature pre B-cell proliferations, appeared at early and intermediate B-cell stages, reached their peak of expression in terms of both incidence of positivity and staining intensity at the late B cell stage (as represented by hairy cell leukemia) and were lost at the B-cell/plasma cell transition. Although detected at similar stages of B-cell differentiation, FMC7 and TracP appear to be independently expressed and were not related to a particular Ig class. The simultaneous detection of FMC7 and TracP represents a distinguishing parameter for the identification of hairy cell leukemia.

Acid Phosphatase↗

Hodgkin's disease-derived cell lines--conflicting clues for the origin of Hodgkin's disease?

The origin of Hodgkin (H) and Reed-Sternberg (RS) cells remains a highly controversial issue. Studies on noncultured, freshly disaggregated biopsy material or of histological sections have not definitely established the cell lineage of HD cells, although evidence has been gathered that has allowed interpretation to favor one or the other hematopoietic cell type. In recent years a number of cell lines have been established from patients with Hodgkin's disease (HD) which are claimed to represent in vitro counterparts of H-RS cells. The phenotypic and functional properties of 7 HD-derived cell lines are reviewed here. The cell lines are not identical; they show many common features, but also a number of important differences in their phenotypes as well as in their in vitro behavior. The cell lines differ with some having characteristics of T cells, others of B cells, and yet others of the monocyte/macrophage cell lineage. This heterogeneity of H-RS established cell lines could be explained in a variety of ways: (a) the heterogeneity of HD itself might result from a disease process which leads to fusion of different cell types resulting in different phenotypic forms of H-RS cells depending on the cells involved; (b) the different cell types might reflect differences in the malignant cells present in the separate subtypes of HD; (c) the difficulty in successfully establishing H-RS cell lines might mean that unrepresentative cell types adapt to in vitro conditions. Rather than resolve the origin of H-RS cells, the established HD-derived cell lines have been consistent with the heterogeneity described in histochemical, immunohistological, and immunocytochemical studies.

Adult↗

Occurrence of particular isoenzymes in fresh and cultured leukemia-lymphoma cells. II. Hexosaminidase I isoenzyme.

The isoenzyme profiles of hexosaminidase (N-acetyl-beta-D-glucosaminidase) were analyzed by isoelectric focusing on horizontal polyacrylamide thin-layer gel with special emphasis on the intermediate isoenzyme (Hex I). The expression of Hex I was examined in 87 leukemia-lymphoma cell lines, in 14 B-lymphoblastoid cell lines, in 441 cases of leukemia-lymphoma (specimens containing 80% or more tumor cells), in 22 leukemia cell lines and in 14 cases of leukemia that had been treated with phorbolesters (TPA) for induction of differentiation, and in the mononuclear cell preparations separated from peripheral blood, lymph node, thymus, bone marrow, tonsil, liver, and spleen specimens from normal donors. Hex I was detected in the leukemia cell lines arrested at early, immature or at late, mature stages of B- and T-cell differentiation, but not in cell lines blocked at intermediate stages of maturation. Most myelomonocytic leukemia cell lines and the erythroleukemia cell lines showed Hex I, whereas the B-lymphoblastoid cell lines were negative for this marker. During induction of differentiation, the expression of Hex I was lost in 13 of 15 leukemia cell lines that were originally Hex I-positive. Among the panel of the "fresh" leukemia-lymphoma cells, Hex I was found predominantly in cases of acute lymphoblastic leukemia and acute myeloblastic/monoblastic leukemia, but rarely or not at all in the mature T-, B- or myeloid malignancies. However, two out of two cases of multiple myeloma were Hex I-positive, and the Hex I expression could be induced by TPA in three of six B-cell chronic lymphocytic leukemia cases. Chronic myelocytic leukemia cells remained Hex I-negative during induction of differentiation. Hex I-positivity was not detected in the cell preparations from normal tissues, and peripheral blood indicating that the normal cellular counterpart of the Hex I-positive tumor cells are present at only low percentages within the respective cell populations. It is suggested that Hex I is a marker of early lymphoid and myeloid hematopoiesis that is no longer expressed in intermediate stages of lymphoid differentiation and in later or terminal stages of myeloid differentiation, but that is again detectable in terminally differentiated B-cells. Further studies will focus on identification and isolation of normal Hex I-positive cells.

Acetylglucosaminidase↗