Search PubMed⌕ Search

Biomedical subjects

J Minowada

Publications and source records attributed to J Minowada.

At least 199 records · Page 11Linked to original sources
PubMed1983

Non-Hodgkin's lymphoma phenotyping: problems in the use of heterologous and monoclonal antibodies.

Cell suspensions or frozen sections of lymph node biopsies from 32 patients with non-Hodgkin's lymphoma (NHL) were studied for sheep erythrocyte (E)-binding under three conditions (Estandard, EAET, Egravity), Fc and C receptors, immunoglobulin (Ig) heavy and light chain class and reactivity with heterologous antisera to T cells (T-LCL), HLA-D (Ia-like) and common acute lymphocytic leukemia (c-ALL) antigens. Selected B and T cell lymphomas were also tested for reactivity with the monoclonal antibodies OKT 3, OKT 4, OKT 6, OKT 8, OKT 11A, Leu-1, Leu-2a, Leu-3a, Leu-4 and Leu-7. There were 26 B and 6 T lymphomas. Most B lymphomas were mu+ (81%), kappa+ (77%) and 31% were mu+ delta+. One of the T lymphomas arose in a patient with antecedent follicular small-cleaved (B) cell lymphoma. The most accurate marker for characterizing the immunologic phenotype in NHL was the clonal excess of kappa+ or lambda+ cells. Neither Estandard, EAET, Egravity or T-LCL were consistently reliable as sole reagents in identifying T-cell lymphomas, their individual scores often being lower than those of monoclonal pan-T cell reagents. HLA-D (Ia-like) antigen was noted in 89% of B and 50% of T lymphomas. The corresponding values for c-ALL antigen were 12 and 33%, respectively. The comparative scores in T-lymphomas between OKT 4 and Leu-3a for "helper-inducer" (HE) cells and OKT 8 and Leu-2a for "suppressor-cytotoxic" (SU) cells were not uniformly consistent. Four T lymphomas had a mixed HE/SU cell phenotype, one was HE, and another SU. Anti-T reactivity was detected in the neoplastic follicles of six of seven follicular lymphomas. The percentage of anti-T reactive cells within positive neoplastic follicles was usually small (5-15%) and of the same order as that noted within reactive lymphoid follicles (5-30%). High numbers (50-100%) of cells from five small lymphocytic B, three diffuse small cleaved cell B and six T cell lymphomas were also positive with one or more anti-T reagents, suggesting the presence of cross-reactive antigens that make phenotyping of lymphomas with monoclonal antibodies problematic. Reactivity with the monoclonal antibody Leu-7 (HNK-1), a putative NK-specific reagent, was seen in one of five B and three of five T lymphomas.

Antibodies, Monoclonal↗
PubMed1983

Terminal transferase immunofluorescence, enzyme markers and immunological profile of human leukemia-lymphoma cell lines representing different levels of differentiation.

We have examined alterations in terminal deoxynucleotidyl transferase (TdT) immunofluorescence (IF) in MOLT-4 cells during changes in growth conditions. Subsequently, we used cells from 48 human hematopoietic cell lines of different cell lineages and maturation stages to compare the IF and biochemical assays for expression of TdT. In addition, we have attempted to correlate the expression of TdT, adenosine deaminase (ADA), thymidine phosphorylase (TP) and immunological markers with maturation stages in these different cell lines. The results indicate that TdT positive cells remain TdT positive when assayed by either the biochemical or IF tests during growth or early plateau phase, but that cells under poor growth conditions, such as in old cultures, may give a negative TdT IF reaction. Otherwise, biochemical and IF assays for TdT gave comparable results in the 48 cell lines tested, testifying to the reliability of the IF test. Based on the comparisons of the various cell lines studied, it appears that both ADA and TdT decrease progressively as maturation of T cells from Blast I to Blast IV to mature T cells increases. TP was deficient in all T-cell lines compared to normal peripheral blood T cells, which in turn had lower activity compared to normal peripheral blood B cells. Pre-B cells, although indistinguishable from each other by immunological markers and all having low TP and ADA activity, showed heterogeneity, with TdT activity high in some and low in others. All non-T, non-B lines had high TdT activity, but low ADA and TP activity. B- and myelocytic cell lines had low ADA and TdT activity, and showed an increase in TP activity as the maturation of cells increased. These results indicate that the TdT IF test is a reliable procedure for detecting TdT positive cells, and that TdT, ADA and TP could be useful markers for studying the differentiation of human hematopoietic cells.

Cell Differentiation↗
PubMed1983

Cytochemical comparison of immunologically characterized human leukaemia/lymphoma cell lines representing different levels of maturation.

Forty-seven human leukaemia/lymphoma cell lines belonging to myelocytic, monocytic, non-T/non-B, T-, and B-lineage and representing different levels of maturation as well as fresh cells from normal and leukaemic subjects were examined for immunological markers and cytochemically for acid phosphatase, alkaline phosphatase, alpha-naphthyl acetate esterase (pH 5.8 and 8.0), alpha-naphthyl butyrate esterase (pH 5.8 and 8.0), non-specific esterase, chloroacetate esterase, chymotrypsin-like protease, deoxyribonuclease II, beta-glucuronidase, sudan black, and periodic acid Schiff's staining. Strong sudan black, nonspecific esterase, and chloroacetate esterase reaction was obtained only for myelocytic and monocytic cell lines with the reaction intensity increasing progressively in more mature cells. Focal acid phosphatase reaction like T-ALL was found in all T-ALL cell lines, whereas myeloid/monocytoid lines had semicircular distribution and B-cell lines cytoplasmic distribution of activity. Acid phosphatase activity appeared to decline with maturation along both myeloid and T-cell lineage. High activity of alpha-naphthyl acetate esterase and alpha-naphthyl butyrate esterase both at pH 5.8 and 8.0 and of beta-glucuronidase was found in myeloid/monocytoid lines although both B- and T-cell lines in contrast to peripheral blood B-cells also had significant esterase activity. alpha-Naphthyl butyrate esterase activity declined with increasing cell maturation along myeloid lineage. Except for weak activity in two B-cell lines alkaline phosphatase was not detected in any cell lines. Monocyte esterase activity was inhibited by sodium fluoride whereas acid phosphatase, only from hairy cell leukaemia line, was resistant to L-tartarate. Although periodic acid Schiff's staining could not distinguish myeloid, T-, B-, or non-T/non-B cell lines it gave characteristic reaction (large number of coarse granules against a clear background forming a ring around the nucleus) with erythroblastic leukaemia cell line and along myeloid series its intensity increased in more mature cells. Deoxyribonuclease II and chymotrypsin-like protease staining were not discriminatory. The results of this study show that cytochemical staining characteristics of various leukaemia/lymphoma cell lines are comparable to those of corresponding cells from patients and that the intensity and pattern of expression of these activities are related to cell type and degree of cell maturation. These studies give further credence to the use of these cell lines in cell differentiation, differential drug cytotoxicity, and many other studies.

Cell Line↗
PubMed1983

Studies of cultured human T lymphocytes. II. Initiation of paired T- and B-cell lines from healthy donors.

We report the sustained cultivation of both B- and T-lymphoblastoid cell lines from randomly selected healthy donors, and the results of studies defining the frequency with which these cell lines can be established. B-cell lines were initiated using the Epstein-Barr virus. Of 52 attempts, 40 B-cell lines (77% success) were obtained from 24 different donors. T-cell lines were started and propagated in long-term (greater than 100 days) cultures using the T-cell growth factor interleukin-2 (IL-2). Of 55 attempts, 54 (98%) were successful in initiating IL-2-dependent T-cell lines, and these were derived from 28 healthy adults. Likewise, of 45 attempts, 32 (71%) were successful in producing paired lines in which both the B-cell line and T-cell line were cultivated from a single blood collection (N = 22 donors). Phenotypic profiles of these lines were defined using multiple marker assays, including rosette formation, surface immunoglobulins, cytochemistry, karyotype, as well as xenoantisera and monoclonal antibodies defining different membrane antigens. This work demonstrates the feasibility of propagating paired human B and T lymphoblastoid lines suitable for many comparative immunobiological studies.

Adolescent↗
PubMed1983

Monoclonal antibody-defined B-cell, T-cell and myelomonocytic antigens and other surface determinants on leukemic B cells of chronic lymphocytic leukemia.

CLL B cells were reactive with monoclonal B-cell antibody (Leu-10) in all patients tested or with monoclonal and heterologous antibodies to Ia-like antigens in all but one patient tested whereas these cells were nonreactive to monoclonal and heterologous antibodies to the cALL antigen in all 34 patients tested. Unexpectedly, of a total of 35 patients with B-CLL, we observed reactivity with monoclonal pan-T antibodies Leu-1, T-101, and OKT-1 (specific for a 65-67,000 dalton surface antigen) in 25 (93%) of 27 patients, in 14 (88%) of 16 patients and in 18 (78%) of 23 patients studied, respectively. Leukemic B cells were reactive with OKT-3 (19,000 dalton) pan-T antibody in 10 (56%) of 18 patients or with Leu-4 (28,000 dalton) pan-T antibody in only 2 (12%) of 17 patients studied. Sheep erythrocyte-rosetting T cells ranged from 0 to 24%. However, a majority of cells (39-100%) reacted with OKT-11 (E-receptor) in 8 (44%) of 18 patients but with Leu-5 (E-receptors) in only 2 (22%) of 9 patients studied. Helper OKT-4 and suppressor OKT-5/8 antibodies were reactive with leukemic B cells in 4 (29%) of 14 patients and in 7 (47%) of 15 patients studied. In contrast, the leukemic B cells from all but one of 22 patients did not react with either helper Leu-3 or suppressor Leu-2 antibody. Unexpectedly, leukemic B cells were also found to be reactive with myeloid or myelomonocytic antibodies (OKM-1 and MCS-1) in 9 (50%) of 18 patients and in 2 (17%) of 12 patients tested. These data suggest that subset of patients with B-CLL demonstrate expression of multiple T and/or myelomonocytic antigens on their leukemic cell populations. Expression of these determinants may reflect biological or clinical significance in CLL.

Antibodies, Monoclonal↗
PubMed1983

Clonal chromosome changes in stimulated lymphocytes of untreated human B-cell leukemias.

The chromosome constitution of peripheral blood lymphocytes from nine cases of B-cell chronic lymphocytic leukemia and one case of B-cell prolymphocytic leukemia were studied at diagnosis following stimulation by B- and T-cell activators. Chromosome analysis with banding techniques revealed an extra chromosome 12 (trisomy 12) in 4 B-cell chronic lymphocytic leukemia cases and complicated abnormalities, i.e., trisomy 12, 6q-, 14q+, and a translocation between chromosomes 6 and 12, [t(6; 12)] in the prolymphocytic leukemia case. These findings suggest that trisomy 12 is a prototypic change in B-cell leukemia, particularly in B-cell chronic lymphocytic leukemia. Based on sister chromatid exchange studies of unstimulated lymphocytes, it appears that some leukemia cells with a normal karyotype not only divide but also proliferate in vitro.

Aged↗
PubMed1983

Possible specific chromosome change in prolymphocytic leukemia.

The chromosomes of unstimulated and stimulated blood lymphocytes from 5 cases with B-cell prolymphocytic leukemia (PLL) were examined following the use of polyclonal B-cell activators (PBA). Banding techniques revealed a common and specific chromosome abnormality to be present in each of the cases, which was due to a translocation between chromosomes 6 and 12 (t(6;12)(q15;p13]. The fact that this specific chromosome change has not been reported in other lymphoproliferative disorders may indicate that PLL is a distinct clinical entity and different from other lymphoproliferative disorders, whether it occurs de novo or complicates chronic lymphocytic leukemia (CLL).

Aged↗
PubMed1982

Urokinase-like plasminogen activators of unusually high molecular weight secreted by a cell line derived from a human lung cancer case.

The HPL-SK-1 cell line derived from the pleural exudate of a lung cancer patient has been shown to secrete plasminogen activators of very high molecular weights (greater than or equal to 2 and 1 million), as shown by gel filtration on Sepharose 6B or CL-6B. The size of these activators could not be reduced by chromatography in buffers containing 2% sodium dodecyl sulfate, 8 M urea, or 1 M KSCN. Goat anti-urokinase antibody inhibited these activators only partially. Trypsin digestion of the 2 million-dalton species yielded several active fragments including one of the size of urokinase, 55,000 daltons. These large activators could be purified only by a double antibody immunoadsorption technique which consisted of the formation of a soluble immune complex between the activators and goat anti-urokinase IgG, followed by the adsorption of this complex to rabbit anti-goat IgG coupled to Affi-Gel 10. The eluted activators were purified 50-fold (2 million daltons) and 130-fold (1 million daltons), respectively. Reduction of the two largest species in the presence of sodium dodecyl sulfate resulted in the appearance of smaller molecular weight active fragments of differing size, indicating that these activators are disulfide-linked oligomers. Among the fragments of the 2 million-dalton species was found a 10,000-dalton enzyme which had lost activator and antigenic specificity and retained only a non-specific protease activity. A similar fragment was also isolated from reduced, purified 55,000-dalton urinary urokinase.

Antigen-Antibody Complex↗
PubMed1982

Modulation of T leukaemic cell phenotype with phorbol ester.

A panel of monoclonal antibodies and other markers (e.g., terminal deoxynucleotidyl transferase, sheep erythrocyte rosettes, peanut agglutinin) have been used in conjunction with flow cytometry and biochemical analysis to monitor the induction of maturation in human thymic (T) leukaemic cell lines by phorbol ester (TPA). Seven cell lines underwent multiple phenotypic alterations in response to TPA but were unresponsive to synthetic thymic hormones (TP5, FTS) or to other compounds (e.g. DMSO, retinoic acid) which induce maturation in other types of leukaemia. The changes parallel those observed in normal T-cell differentiation and partly reflect alterations in glycosyl transferase activity, altered synthesis of proteins and regulation of cell surface receptors (for transferrin) associated with rapid growth and metabolism. These studies further illustrate the reversibility of maturation arrest in human leukaemia and provide support for the view that leukaemia may involve regulatory defects in the coupling of proliferation and maturation. Induction of promotion of terminal differentiation in leukaemic equivalents of T-cell precursors may provide a convenient system for the study of biochemical and molecular events involved in T-cell development and diversification.

Antibodies, Monoclonal↗
PubMed1982

Studies of cultured human T lymphocytes. I. Production of the T cell growth-promoting lymphokine interleukin-2.

Described herein is a large-scale procedure that has been successfully employed for producing 62 lots (800-3000 ml) of supernatants containing the T cell growth-promoting factor Interleukin-2 (IL-2). The efficiency of these crude, unconcentrated supernatants was documented in studies in which 70 human long-term (greater than 100 days) IL-2-dependent T cell lines were established from 50 different donors. These included lines initiated from the peripheral blood of healthy subjects (N = 54), blood of children with active acute lymphoblastic leukemia (N = 6) and the thymus of children undergoing surgery to correct congenital heart defects (N = 10). The underlying concept used in constructing this method emphasizes the requirement of the monocyte-derived macrophage and its Interleukin-1 (IL-1) product to mediate IL-2 production by activated T cells. The most salient feature of this technique is the utilization of buffy coat leukocytes that had been pooled from several blood donors and sustained in spinner cultures for several days prior to polyclonal activation with phytohemagglutinin and pooled B cells of established human lymphoblastoid lines.

Adolescent↗
PubMed1982

Analysis of human leukemia/lymphoma cell lines with monoclonal antibodies BA-1, BA-2 and BA-3.

A panel of monoclonal antibodies (BA-1, BA-2 and BA-3) were made against the pre-B acute lymphoblastic leukemia (ALL) cell line NALM-6. BA-3 binds to the 100,000 mol. wt common ALL antigen (gp100/CALLA), BA-2 binds to a 24,000 mol, wt (p24) cell surface structure and BA-1 binds to a currently undefined antigen. These antibodies were analysed for their reactivity with 67 human leukemia/lymphoma cell lines. The cell lines studied included T ALL, non-T ALL, surface immunoglobulin+ leukemias/lymphomas, and myeloid/monocytoid leukemias. The results indicate that all three antibodies react primarily (but not exclusively) with malignant cells early in B cell development. This pattern of reactivity was similar to previous results obtained with fresh, non-cultured malignant cells. BA-1, BA-2 and BA-3 are useful tools for analysing the developmental options of normal lymphoid cells and for "cleaning up" leukemia/lymphoma cells in selected cases of autologous bone marrow transplantation.

Antibodies, Monoclonal↗
PubMed1982

Chromosomes and causation of human cancer and leukemia. XLVIII. T-cell acute leukemia in ataxia telangiectasia.

Cytogenetic and immunologic studies were performed on the cells of an 18-year-old female with ataxia telangiectasia (AT) associated with acute lymphocytic leukemia (ALL). At the onset of the leukemia 15.4% of peripheral blood cells stimulated with phytohemagglutinin (PHA) contained a tandem translocation of the long arm of chromosome #14, i.e., t(14;14). To ascertain if these karyotypically abnormal cells and the leukemic cells had a common lineage, chromosome analyses were performed on bone marrow cells. Examination of the marrow cells on the seven occasions when leukemic cells were present in the marrow, including times when they were predominant, showed only a normal karyotype without the presence of t(14;14). However, an abnormal clone, which had the karyotype 45,XX,-9,t(9;6)(q12;q13), was identified in the marrow cells on the last examination during the terminal phase of the leukemia. Immunologically, the ALL was classified as an atypical type which had characteristics in common with certain T-cell subsets. We suggest that the malignant cells did not originate from the preexisting cells with a tandem duplication of the 14q.

Adolescent↗
PubMed1982

Purification of a lymphoid cell line product with leukocyte inhibitory factor activity.

Leukocyte inhibitory factor (LIF), a lymphokine that inhibits the random and directed migration of polymorphonuclear (PMN) leukocytes, was purified from a human non-T, non-B leukemia cell line (Reh). From 10 liters of serum-free supernatant, 1.3 microgram of protein with LIF activity was obtained by the sequential use of affinity chromatography with concanavalin A-Sepharose, hydrophobic chromatography with hexylagarose, and gel filtration chromatography. The specific activity of LIF recovered represented an 80,000-fold purification over that of the initial crude serum-free supernatants, and the preparation at that point was estimated to be 80--90% pure. To both assess the purity of the preparation and provide a further purification step, Reh LIF activity recovered by the above procedures was subjected to isoelectric focusing. One major stainable protein band was identified; its isoelectric point was pH 5.4--5.5. Gels run in parallel for recovery of biologic activity revealed only one region (pH 5.4--5.5) with ability to inhibit PMN leukocyte migration. Iodination of Reh LIF resulted in a loss of biologic activity, but isoelectric focusing of this material revealed one major 125I-labeled band (pH 5.1) and several minor bands. The coincidence of biologic LIF activity with one stainable protein band as identified by isoelectric focusing implies that the final product may be homogeneous.

Cell Line↗
PubMed1982

Effect of dexamethasone on the growth of human lymphoblastoid cell lines.

Effect of dexamethasone of the growth of two T-cell acute lymphoblastic leukemia (T-ALL) cell lines, JM and TALL-1 and an American Burkitt lymphoma cell line, Ramos, of B-cell type was examined. While the growth of both TALL-1 and Ramos cell lines was significantly inhibited by dexamethasone at the concentration of 10(-7) M, JM cell line was markedly resistant at the high concentration of 10(-5) M of dexamethasone. The assay of glucocorticoid receptors showed that a small amount of the binding site was found in both T-cell leukemia cell lines, JM and TALL-1 and that a moderate amount of the binding site was found in Ramos cell line. Thus, glucocorticoid sensitivity (growth inhibition of these leukemia-lymphoma cell lines by dexamethasone) did not correlate with either the number of glucocorticoid receptor or with surface membrane markers of the target cells. This result suggests further marked heterogeneity of leukemic cells.

Burkitt Lymphoma↗