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J Minowada

Publications and source records attributed to J Minowada.

At least 145 records · Page 8Linked to original sources
PubMed1986

Changes in isoenzyme profiles during induction of differentiation in human myelomonocytic leukemia cell lines.

The human leukemia cell lines HL-60, KG-1, KLM-2, ML-3, THP-1, and U-937 were treated with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). TPA partially or completely inhibited the proliferative activity of the cell cultures. The number of cells with the ability to reduce nitroblue tetrazolium increased in the TPA-treated cell lines HL-60, ML-3, THP-1, and U-937, whereas the cell lines KG-1 and KLM-2 remained nitroblue tetrazolium negative. Except for KG-1 and KLM-2, all TPA-treated cell lines showed varying degrees of strong adherence to plastic surface. The carboxylic esterase, acid phosphatase, hexosaminidase, and lactate dehydrogenase isoenzyme profiles from these cell lines were analyzed by isoelectric focusing on horizontal polyacrylamide gels. The new or stronger expression of an esterase isoenzyme which is specific for monocytes-macrophages was induced in HL-60, ML-3, THP-1, and U-937 but not in KG-1 or KLM-2. The new expression of the tartrate-resistant acid phosphatase isoenzyme was induced in ML-3, THP-1, and U-937. The number of esterase and acid phosphatase isoenzymes and the staining intensity of isoenzymes characteristic for myeloid cells were increased by TPA in all cell lines. The loss of the hexosaminidase I isoenzyme which is a marker of immature hematopoietic cells was noted in KG-1, ML-3, THP-1, and U-937. TPA triggered an increase in number and staining intensity of the lactate dehydrogenase isoenzymes in all cell lines. Some isoenzymatic changes (e.g., monocyte-specific esterase, tartrate-resistant acid phosphatase, hexosaminidase I) appear to correlate with TPA-induced differentiation while other alterations in the isoenzyme patterns do not (e.g., lactate dehydrogenase, other esterase and acid phosphatase isoenzymes). Differentiation of nonmonocytoid cells appears, at the isoenzyme level, to be quite different from that of the monocytoid cell lines.

Acid Phosphatase↗
PubMed1986

Organization of the T-cell receptor alpha-chain gene and rearrangement in human T-cell leukaemias.

The structure and rearrangement of the human T-cell receptor alpha-chain gene have been analysed. The constant region segment is comprised of four exons, with the 3' non-coding region present as a separate exon. Two J alpha segments have been identified by sequence analysis about 4 X 10(3) and 15 X 10(3) base-pairs upstream from the constant region segment. Analysis of alpha-chain rearrangements in a panel of T-cells reveals the presence of at least four more J alpha segments within 19 X 10(3) base-pairs of C alpha and suggests the existence of additional J alpha segments further upstream. This dispersed organization of the J alpha region contrasts with the clustered organization observed in other lymphoid rearranging genes. The use of J alpha probes in diagnostic purposes for T-cell leukaemia is not, therefore, convenient and will require a number of different probes to ensure complete analysis of the locus.

Base Sequence↗
PubMed1986

Monocytoid leukemia cell line CTV-1: morphological, immunological and isoenzymatic characteristics.

The human leukemia cell line CTV-1 was established from a case of acute monoblastic leukemia (AMoL). We analyzed the phenotypic marker profile of the CTV-1 cells in their original, untreated state and during induction of differentiation with the phorbolester 12-0-tetradecanoylphorbol 13-acetate (TPA). TPA led to morphological changes with signs of differentiation. Cell proliferation decreased in a dose-dependent fashion during exposure to TPA. In the surface marker analysis using a panel of 45 monoclonal antibodies (MoAbs) and several polyclonal antisera, CTV-1 cells were negative for markers of the T- and B-cell lineages, and were positive for several, but not all, myelomonocytic markers. Although the cells were reactive with the MoAb Leu-7 which identifies natural killer (NK) T-cells, no NK activity was detected. In the isoenzyme analysis of the four enzymes carboxylic esterase, acid phosphatase, hexosaminidase and lactate dehydrogenase (LDH) performed by isoelectric focusing on polyacrylamide gels, CTV-1 cells displayed isoenzyme profiles of immature myeloid cells. The overall marker profile of CTV-1 cells demonstrated cells of monocytoid origin arrested at a very early stage of differentiation, possibly close to the stage of precursor cells. As compared to other myelomonocytic cell lines, CTV-1 cells showed unusual morphological, immunological, functional and biochemical features and appeared to be relatively insensitive to treatment with TPA, although some alterations of the phenotype could be induced.

Acid Phosphatase↗
PubMed1986

Correlation of surface marker expression with morphologically and immunologically defined subclasses of acute myeloid leukaemias.

The expression of myeloid-associated cell surface antigens detected by monoclonal antibodies (MoAb: MCS-2, MCS-1, MY7, MY9, Leu-M1, OKM1, VIM-D5, Mol, My-1, MY8, MY4, Leu-M3, VIM-D2, Mo2) of the HLA-DR/Ia-like antigen and of the Fc-receptor was determined on the blast cells from 91 patients with acute myeloid leukaemias classified as M1-M5 in the French-American-British (FAB) system. The surface antigen analysis revealed a highly heterogeneous reaction profile. Nevertheless, distinctive patterns of marker expression referring to morphologically defined subgroups were delineated. Several MoAb (especially MCS-2 and MY7 which were positive in most cases of the five FAB subgroups) appear to be useful for the recognition of myelomonocytic cells regardless of the commitment to either the granulocytic or monocytic cell lineage whereas other Mo Ab (especially MY4, Leu-M3, VIM-D2, Mo2) react predominantly with the monocytic variants and are helpful in the identification of monocytic commitment. The 91 cases could be divided into three immunologically defined phenotypes (Types I-III) corresponding to sequential differentiation levels. Correlations of these MoAb-defined phenotypes with the FAB subtyping showed that immunological and morphological classifications are not completely concordant and that only the parameters Type I and FAB M1 were significantly related. A scheme of early myeloid differentiation sequences based on the expression of surface antigens is presented.

Adult↗
PubMed1986

Pre-B and non-T/non-B leukemia cell lines BV-173, KM-3, NALM-6, and REH: changes in isoenzyme profiles during induction of differentiation.

The ability of 12-O-tetradecanoylphorbol 13-acetate (TPA) to induce stable phenotypic changes that serve as markers of differentiation was examined in the non-T/non-B leukemia cell lines KM-3 and REH and in the pre-B leukemia cell lines BV-173 and NALM-6. Isoenzymes of the enzymes carboxylic esterase, acid phosphatase, hexosaminidase, and lactate dehydrogenase (LDH), separated by isoelectric focusing on horizontal polyacrylamide thin-layer gels, were used to monitor induced changes. TPA in different concentrations completely or partially inhibited cell proliferation, but had no drug-related cytotoxicity. No increase in the number of nitro-blue-tetrazolium-reducing cells nor adherence to plastic surface was found. In all four cell lines, TPA caused an increase in number and staining intensity of esterase, acid phosphatase, and LDH isoenzymes. The resulting isoenzyme profiles corresponded to those seen at more mature intermediate stages of B-cell proliferation, but did not indicate a terminal differentiation to mature B cells. The loss of the hexosaminidase I isoenzyme, which is a marker of immature hematopoietic cells, was a further indicator of induced maturation. These results demonstrate that while TPA is capable of inducing various immature non-T/non-B and pre-B cell lines to differentiate, the differentiation progression appears to be restricted to intermediate stages, in contrast to the terminal differentiation inducible in myeloid cells.

Acetylglucosaminidase↗
PubMed1985

Burkitt cell acute lymphoblastic leukemia with partial expression of T-cell markers and subclonal chromosome abnormalities in a man with acquired immunodeficiency syndrome.

A 45-year-old white male, bisexual, with a 2-year history of acquired immunodeficiency syndrome (AIDS) prodrome, developed a Burkitt cell-like acute lymphoblastic leukemia (ALL). Marker studies of marrow blasts show an unusual and possibly unique pattern, in that an unequivocal monoclonal B cell leukemia, having K-IgM with HLA-DR and B cell subset antigen (BA-1) expression, was superimposed with a mature suppressor T cell marker profile (pan-T, mature T, and suppressor/cytotoxic T antigens). The leukemic blasts were totally negative for terminal deoxynucleotidyl transferase (TdT), human T cell leukemia-lymphoma virus (HTLV) p19 antigen, and other immunoglobulin isotypes. Chromosome analysis of marrow cells disclosed that 70% of the cells had 47,XY, + 12,t(8;14)(q24;q32) chromosome complement, and 30% of the cells had a 47,XY, + 12,dup1q + (q22-31),t(8;14)(q24;q32). The consistent finding of the specific chromosome rearrangement (8/14 translocation) in all abnormal cells suggests that the cells were derived from a common precursor. With regard to the partial T cell marker expression, the significance of these markers in B cell leukemia is unclear, as is their relation to the additional chromosome abnormalities that apparently developed in the process of clonal evolution.

Acquired Immunodeficiency Syndrome↗
PubMed1985

Cytogenetic studies of a diffuse mixed cell lymphoma of T cell origin.

The chromosome finding obtained from a lymph node of a patient with T cell lymphoma is described. Two different abnormal clones were found; one of them had a t(14;18)(q32;q21), along with other structural and numerical abnormalities, including 1p+q-,1p-,2p+q+, der(11),t(11;?)(q13;?),+20,+21,22p+. The other clone contained der(13),t(13;?)(q22;?).

Adolescent↗
PubMed1985

High concordance between marker profiles of 22 human leukemia-lymphoma cell lines tested with the same monoclonal antibodies before and during the second international workshop on human differentiation antigens.

Our laboratory participated in the Second International Workshop and Conference on Human Leucocyte Differentiation Antigens. In this international study the reactivity profiles of monoclonal antibodies were analyzed on normal and malignant hematopoietic cells. The Workshop was divided into three categories: the T-cell, B-cell and myelomonocytic cell studies. We blindly tested 159 coded monoclonal antibodies of the panel for the T-cell study on 22 permanently established leukemia cell lines. The monoclonal antibodies were provided by the Workshop Committee and their reactivity with the target cells was visualized by standardized indirect immunofluorescence. After decoding it was recognized that 11 monoclonal antibodies had been examined on these cell lines prior to the Workshop. The reactivity of these 11 monoclonal antibodies was analyzed and compared with the earlier results. From a total of 217 paired tests done blindly in the Workshop study and prior to the Workshop, 191 tests (88%) did not show significantly different data. The possible reasons for discrepancies include nonspecific Fc-receptor-binding on some cell lines and a relatively nonspecific reactivity of some monoclonal antibodies. This analysis demonstrates the stability of the antigen expression on human leukemia-lymphoma cell lines grown at consistently optimal conditions, for the tests, using the same monoclonal antibodies as in the Workshop, had been performed 0.5-5 years prior to the Workshop study. On the other hand, nonspecific Fc-binding, wide "specificity" of monoclonal antibodies and a shift in antigen expression of the cells (due to poor growth conditions, involuntary induction of differentiation and other factors) must be taken into consideration upon immunological analysis.

Antibodies, Monoclonal↗
PubMed1985

Isoenzyme studies in human leukemia-lymphoma cell lines--1. Carboxylic esterase.

The isoenzyme patterns of carboxylic esterase (E.C. 3.1.1.1) were studied in 74 proven human leukemia-lymphoma and 12 normal B-lymphoblastoid cell lines. These cell lines have been extensively phenotyped using poly- and monoclonal antibodies. Esterase isoenzymes were separated by isoelectric focusing and visualized by histo-cytochemical techniques. No leukemia-specific or (except for monocytes) blood cell type-specific isoenzyme or isoenzyme pattern could be detected. The monocytic element in some cell lines was characterized by a strong isoenzyme band which could be selectively and completely inhibited by sodium fluoride. The enzyme phenotypes were stably expressed in all subcultures of a given cell line and did not appear to have any cell cycle dependency. The leukemia-lymphoma cell lines have been subclassified into four major groups according to immunological parameters: T-cell, B-cell, myelomonocytic and non-T, non-B-cell. On the basis of immunological data the T-cell lines were assigned to five stages of differentiation. The number and staining intensity of the isoenzymes increased with differentiation of the T-cells paralleling the expression of immunological markers. The B-cell leukemia-lymphoma cell lines were divided into pre B-, B-, Burkitt lymphoma, multiple myeloma and hairy cell leukemia cell lines. Substantial variability among the isoenzyme patterns was detected ranging from immature profiles of pre B-cell lines to complete isoenzyme repertoires of multiple myeloma cell lines. No significant difference was seen between the isoenzymes of mature B-cell lines and normal B-lymphoblastoid cell lines. The most prominent feature seen in myelomonocytic cell lines was the monocytic band indicating a monocytic origin and separating the 'monocytoid' from the 'pure myeloid' cell lines. Considerable heterogeneity in the isoenzyme patterns was observed in the non-T, non-B cell groups which comprised erythroleukemia cell lines and cell lines arrested at a very early stage of lymphoid differentiation. These latter cell lines together with some T- and B-cell lines shared the common characteristics of positivity for cALLA, TdT and Ia antigens and an immature, incomplete isoenzyme profile. The results support the notions of maturation arrest and normal gene expression in leukemic cell populations. Furthermore, the importance of biochemical studies as part of the multiple marker analysis could be demonstrated.

Animals↗
PubMed1985

Catalogue of Epstein-Barr virus (EBV) receptors on human malignant and non-malignant hematopoietic cell lines.

Epstein-Barr virus (EBV) can induce a broad spectrum of hematological diseases, especially in immune deficient patients. We assayed for receptor for EBV (EBVR) using fluoresceinated viral particles on 44 human hematopoietic cell lines derived from patients with T, B, and non-T, non-B acute lymphocytic leukemia (ALL), non-lymphoid leukemia, Burkitt lymphoma, myeloma and several unique lines we and others have recently developed. All 31 EBV nuclear-associated antigen (EBNA) negative cell lines were of neoplastic origin. Seven of 13 EBNA-positive cell lines were of normal cell origin. Four of 25 non-B (surface immunoglobulin negative) EBNA-negative neoplastic cell lines were EBVR-positive. Three of six EBNA-negative B-cell (surface immunoglobulin positive) lines were EBVR-positive. Nine of 13 EBNA-positive Burkitt and non-Burkitt cell lines strongly expressed EBVR. Four EBNA-positive Burkitt lymphoma cell lines exhibited EBVR only to a limited degree. Studies of the cell lines for EBVR, complement receptors (CR) and surface immunoglobulin (SIg) revealed that presence of SIg does not obligate the presence of EBVR. Functional EBVR accompanied SIg among EBNA-negative cell lines. SIg-negative cell lines can possess EBVR. Fourteen of 16 EBVR-positive lines were also positive for CR. The EBVR assay is a useful tool for assessing the potential role of EBV in the induction of hematopoietic disorders.

Antigens, Surface↗
PubMed1985

Isoenzyme studies in human leukemia-lymphoma cells lines--II. Acid phosphatase.

This report describes the qualitative acid phosphatase (acP) isoenzyme profiles detected in permanent human hematopoietic cell lines. The acP activity was separated into its isoenzymes by isoelectric focusing on horizontal thin-layer polyacrylamide gels. The pattern of acP isoenzyme was investigated in a total of 86 cell lines. These cell lines were classified into five groups on the basis of their phenotypes characterized in the multiple marker analysis: 74 leukemia-lymphoma cell lines (26 T-, 34 B-, 6 myelomonocytic, 8 Non-T, Non-B cell lines) and 12 so-called 'normal' Epstein-Barr virus transformed B-lymphoblastoid cell lines. Their immunological features had been analysed in detail by use of a large panel of poly- and monoclonal antibodies which led to a further subclassification into stages of differentiation. A progressive increase in number and staining intensity of the isoenzymes which paralleled the expression of surface markers at different stages of differentiation along their developmental pathway was seen in the T- and B-leukemia-lymphoma cell lines. Some cell lines whose isoenzyme profiles did not correspond to the stage of differentiation as evidenced by surface antigen analysis might represent good examples of deranged gene expression in otherwise normally programmed malignant cells, i.e. in our study a mismatch between the isoenzymatic and immunological phenotypes. The tartrate-resistant isoenzyme was detected in 9 out of 74 leukemia-lymphoma cell lines (4 T-, 2 B-, 1 myelomonocytic, 2 Non-T, Non-B cell lines) and in 10 out of 12 normal B-lymphoblastoid cell lines; the only one studied hairy cell leukemia cell line did not express this isoenzyme. The relative specificity of the tartrate-resistant acP is discussed in detail. No leukemia-lymphoma specific isoenzyme or an additional isoenzyme which was not seen in normal hematopoietic cells could be observed. Nor did we find an isoenzyme or isoenzyme pattern characteristic for a certain cell lineage. This underlines the necessity of a combined analysis using markers from different disciplines in the 'multiple marker analysis' in order to accurately characterize normal and malignant blood cells. Furthermore, our results support the concept of maturation arrest at particular stages of differentiation together with the theory of normal gene expression in leukemic cells equivalent to that in their normal counterparts.

Acid Phosphatase↗
PubMed1985

Isoenzyme studies in human leukemia-lymphoma cell lines--III. beta-Hexosaminidase (E.C. 3.2.1.30).

The hexosaminidase (beta-N-acetylglucosaminidase) isoenzyme profiles of 86 human hematopoietic cell lines grown actively in suspension culture were analysed by isoelectric focusing and by conventional disc electrophoresis on horizontal thin-layer polyacrylamide gels. A maximum of three hexosaminidase (Hex) isoenzymes (A = anodic, I = intermediate, B = basic) could be demonstrated. The immunological phenotyping of 74 leukemia-lymphoma derived cell lines had led to a categorization into four groups with a subclassification of the T- and B-cell lines into several stages of differentiation: 26 T-cell, 34 B-cell, 6 myelomonocytic and 8 Non-T, Non-B cell leukemia-lymphoma cell lines. Twelve so-called 'normal' B-lymphoblastoid cell lines were also available. Distinct isoenzyme profiles were seen in the different stages of differentiation in the T- and B-leukemia-lymphoma cell lines. Among the 12 normal B-lymphoblastoid cell lines heterogeneity in the isoenzymatic phenotypes was detected. Hex isoenzyme expression in normal and neoplastic lymphoid cell lines represents hypothetically sequential stages of T- and B-cell differentiation. Myelomonocytic cell lines displayed strongly stained bands of all three isoenzymes. Heterogeneity was seen in the group of Non-T, Non-B cell lines. Four out of 5 pre B-cell lines and 4 out of 4 Non-T, Non-B cell lines which are comparable to cases of pre B- and common ALL revealed a high Hex I/Hex A ratio in terms of intensity of the isoenzyme bands. The analysis of Hex isoenzymes is useful for characterizing lymphoid and myeloid populations (both normal and malignant, cultured or fresh), particularly with regard to their stage of differentiation. But this enzyme should be part of a multiple enzyme study where the information obtained is complementary. In turn, enzyme marker analysis should be included in the multiple marker analysis for an optimized characterization of leukemic cells.

Antigens, Surface↗
PubMed1985

Isoenzyme studies in human leukemia-lymphoma cell lines--IV. Lactate dehydrogenase.

Isoelectric focusing (IEF) in horizontal polyacrylamide gels has been used to separate lactate dehydrogenase (LDH) isoenzymes in 97 human permanent hematopoietic cell lines (85 leukemia-lymphoma cell lines and 12 'normal' B-lymphoblastoid cell lines). Maximally 8 LDH bands were seen; the electrophoretically detectable bands 4 and 5 could be separated by IEF into 2 and 3 isoenzymes, respectively. The LDH patterns have been found to vary both in number of isoenzymes and in relative intensity in different cell lines depending upon the stage at which arrest of differentiation occurred. These differences can be used to analyse and distinguish different cell lines. The method should provide a valuable supplement to the enzymatic phenotyping and complete characterization of fresh and cultured leukemias and for the monitoring of phenotypic changes occurring during induction of differentiation.

Antigens, Surface↗
PubMed1985

Expression of a monocyte-specific esterase isoenzyme in cases of acute myeloid leukemias.

The carboxylic esterase (E.C. 3.1.1.1) isoenzymes from cases of acute myeloid leukemias were separated by analytical isoelectric focusing on horizontal thin-layer gels. One isoenzyme consisting of one or two components (bands) could be completely and selectively inhibited by addition of 40 mM sodium fluoride (NaF) to the staining bath. The 105 cases were classified into the groups M1-M6 according to the FAB proposals. The NaF-sensitive isoenzyme was not detected in cases of FAB groups M1/2 (acute myeloblastic leukemia without or with maturation), group M3 (acute promyelocytic leukemia) or group M6 (erythroleukemia). Thirty-one out of 33 cases in the FAB group M4 (acute myelomonocytic leukemia) and 9/9 cases in FAB group M5 (acute monocytic leukemia) expressed the NaF-sensitive isoenzyme. The NaF-sensitive isoenzyme was found at different staining intensities; all M5 cases showed the isoenzyme at strong or very strong intensity, whereas most of the M4 cases displayed the isoenzyme at weak, medium or strong staining intensity. The data presented are further evidence that the presence of the NaF-sensitive esterase isoenzyme indicates monocytic involvement or differentiation in cases of myeloid leukemias. The easy and fast to perform method of isoelectric focusing can be used to distinguish the monocytic variants among the acute myeloid leukemias and can supplement the morphological analysis of these cases.

Carboxylic Ester Hydrolases↗
PubMed1985

Biochemical enzyme analysis in acute leukaemia.

This report summarises the current knowledge regarding the clinical utility of biochemical enzyme markers for both diagnostic and therapeutic purposes in acute leukaemia. The enzymes studied most extensively in this field are terminal deoxynucleotidyl transferase, adenosine deaminase, 5'-nucleotidase, purine nucleoside phosphorylase, and acid phosphatase, esterase, hexosaminidase isoenzymes. For each enzyme, the quantitative and qualitative characteristics in various immunologically defined subclasses of acute leukaemia are described. The quantitative evaluation of enzyme activities represents an adjunctive classification technique which should be incorporated into the multivariate analysis, the "multiple marker analysis." By qualitative characterisation pronounced heterogeneity of leukaemia subsets is uncovered. The application of 2'-deoxycoformycin, a specific inhibitor of adenosine deaminase, and the potential usefulness of two other enzymes as targets for treatment with selective agents is discussed. The concept that gene products expressed at certain developmental stages of normal cells can similarly be detected in leukaemic cells (which therefore seem to be "frozen" or "arrested" at this particular maturation/differentiation stage) is supported by the results obtained in enzyme studies. Besides their practical clinical importance for classification and treatment of acute leukaemias, biochemical enzyme markers constitute a valuable research tool to disclose biological properties of leukaemic cells.

5'-Nucleotidase↗
PubMed1985

Subsets of normal mononuclear cells in the peripheral blood defined by monoclonal antibodies.

We studied the peripheral blood of 20 healthy adult individuals for the distribution of subsets within the mononuclear cell population using indirect immunofluorescence staining. Normal ranges are given for several cell populations reacting with a panel of monoclonal antibodies. The analyzed immunological reagents included the monoclonal antibodies Leu-1 (Pan-T), Leu-2a (Suppressor-T), Leu-3a (Helper-T), Leu-7 (Natural Killer-T), MCS-2 (Pan-myeloid), NU-B1 (Pan-B), FMC-7 (B-subset), Tac (IL-2-receptor) and sheep erythrocyte rosetting (E). Cells were examined for positivity under an epi-immunofluorescence microscope and by flow cytometry. The data obtained by these two different approaches were in close agreement. The majority of the cells from the total mononuclear cell fraction in the peripheral blood were Leu-1+ (mean value of 64% by microscope/59% by flow cytometry) and E+ (61%). The T-populations could be broken down into Leu-3a+ (48%/43%), Leu-2a+ (31%/31%) and Leu-7+ (13%/14%). Between 2 and 5% of the cells showed double expression of antigens detected by Leu-2a and Leu-3a. The pan-myeloid reagent MCS-2 was positive on 25%/18% of the total cell population; the B-cells accounted for 5%/8%. Tac stained 7%/6% of the cells.

Adult↗