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Biomedical subjects

J Minowada

Publications and source records attributed to J Minowada.

At least 91 records · Page 5Linked to original sources
PubMed1990

CD28 molecule as a receptor-like function for accessory signals in cell-mediated augmentation of IL-2 production.

IL-2 production by PHA-stimulated MOLT 14 cells (a TcR gamma/delta-bearing human leukemic T cell line) and MOLT 16 cells (a TcR alpha/beta-bearing human leukemic T cell line) was markedly augmented by coculturing with BALL-1 cells ( a human leukemic B cell line), or with recombinant human interleukin-1 alpha (rhIL-1 alpha). We have previously shown that the augmentation of IL-2 production, induced by BALL-1 cells, requires cell to cell contact and is an IL-1-independent pathway. In this report, the expression of the CD28 molecule on MOLT 14 cells and MOLT 16 cells was examined for its role in IL-2 production augmented by BALL-1 cells. A 1-hr preincubation of MOLT 14 cells and MOLT 16 cells with anti-CD28 mAb resulted in the inhibition of BALL-1 cell-induced augmentation of IL-2 production (90 and 62% inhibition of control, respectively). The inhibition was observed in a dose-dependent manner of anti-CD28 mAb added and reached a plateau level at concentrations of 0.05 micrograms/ml of anti-CD28 mAb. This was sufficient to cover all the CD28 molecules expressed on the surface of both T cells as detected by flow cytometric analysis. Flow cytometric analysis also showed that the inhibition was not due to a modulation of CD28 molecules. In contrast, the treatment with anti-CD28 mAb did not inhibit IL-2 production which was augmented by rhIL-1 alpha costimulator. These results suggest that the CD28 molecule on the T cells is important for the interaction with BALL-1 cells which causes the augmentation of IL-2 production and further imply that the CD28 molecule is a receptor for an accessory signal provided by BALL-1 cells.

Antibodies, Monoclonal↗
PubMed1990

Polyadenylic acid polymerase activity in chronic myelogenous leukemia.

Poly(A) polymerase activity was markedly elevated in CML in the blastic phase, moderately high in the accelerated phase and low in the chronic phase. The activity was significantly higher in the myeloid crisis than in the lymphoid crisis and elevated with increasing ratio of blasts in leukemia cases. In TPA or retinoic acid-treated leukemia cells poly(A) polymerase activity was decreased. These results suggest that poly(A) polymerase activity changes, depending on the maturation of leukemic cells and the assay of this enzyme activity may be useful for the early detection of the exacerbation of CML cases.

Blast Crisis↗
PubMed1990

A novel CEA-cross-reacting antigen of molecular weight 140,000 expressed on human lymphoid cell lines.

To investigate the possible expression of the carcinoembryonic antigen (CEA) gene family products on lymphoid cells, we screened 28 human cell lines derived from malignant lymphoid cells for reactivity with monoclonal antibodies (MAbs) against CEA and nonspecific cross-reacting antigen (NCA), which is one of the CEA gene family members. Six cell lines (four B cell lines and two non-T, non-B cell lines) were found to react, by a membrane immunofluorescence test, with an MAb, F34-187, which recognizes an antigenic determinant shared between CEA and NCA. None of the 15T cell lines was reactive with any MAbs tested. A glycoprotein antigen isolated with F34-187 from the cell surface showed an apparent molecular mass of ca 140 and 70 kDa in the glycosylated and deglycosylated forms, respectively, and was unreactive with MAbs specific for CEA or NCA, suggesting that the antigen is a new member of the CEA gene family.

Antibodies, Monoclonal↗
PubMed1990

Levels of sister-chromatid exchanges in hybrids between Bloom syndrome B-lymphoblastoid cells and various cell lines with lymphoid malignancy.

The present study has been undertaken to examine the effect of cell hybridization of Bloom syndrome (BS) B-lymphoblastoid cell lines (LCLs) and various cell lines from lymphoid malignancies in order to clarify the relationship between sister-chromatoid exchange (SCE) and malignant conditions. Cell hybridization studies have shown that though BS high-SCE frequencies were completed by fusion with normal cells, fusion with various malignant cell lines did not result in complete normalization of BS SCEs, with 15-30 SCEs remaining per hybrid cell, demonstrating possibly common defects in DNA of BS and malignant cells. These findings strongly support the idea that the characteristic high SCE frequency in BS cells has some connection with the malignant condition, and that at least one step in carcinogenesis is either accompanied by the production of SCEs, or that SCEs themselves cause such a step to occur.

B-Lymphocytes↗
PubMed1990

Low dose oral alpha-interferon therapy for patients seropositive for human immunodeficiency virus type-1 (HIV-1).

Thirty eight symptomatic and two asymptomatic patients seropositive for human immunodeficiency virus type-1 (HIV-1) were treated with a natural human interferon alpha (HuIFN alpha). Patients were given 2 IU/kg HuIFN alpha orally once daily in powdered maltose held in the mouth to promote mucosal absorption. This oral immunomodulating HuIFN alpha therapy resulted in an increase in CD4+ lymphocytes, an increase in weight, and a dramatic alleviation of clinical symptoms related to HIV-1 infection.

Administration, Oral↗
PubMed1990

[T cell differentiation model based on the expression of T cell receptor chains and genes--study in the human leukemia/lymphoma cell lines].

A total of 33 human leukemia/lymphoma cell lines were classified into 4 groups with respect to the pattern of cell membrane (sm) expression of the CD3 and T cell receptor (TCR) molecules; (i) smCD3+TCR alpha beta (16 cell lines), (ii) smCD3+TCR beta delta (1 cell line), (iii) smCD3+TCR gamma delta (3 cell lines) amd (iv) smCD3-TCR- (13 cell lines), respectively. Using monoclonal antibodies (MoAbs) specific to CD3 (NU-T3), TCR alpha chain (alpha F1), TCR beta chain (beta F1), and TCR gamma chain (C gamma M1), respectively, cytoplasmic (cy) expression of these molecules was determined by immunofluorescence test. Expression of cyCD3 was present in all cell lines regardless of groups. In group (i), all 16 cell lines expressed both TCR alpha and beta chains. While only TCR beta chain was expressed in group (ii), TCR gamma chain was expressed in all 3 cell lines of group (iii). One (PEER) of the three in group (iii) expressed TCR beta chain as well. In group (iv), we found 8 cell lines with cyTCR alpha expression, 11 cell lines with cyTCR beta expression, and 10 cell lines with cyTCR gamma expression, respectively. For TCR genes, except 1 cell line all cell lines were found to present rearranged C beta gene and its mRNA, including all 3 TCR gamma/delta cell lines of group (iii). One of the TCR alpha beta cell lines exhibited rearranged C delta and J delta genes as well as its mRNA. Two cell lines of the 13 CD3-TCR- of group (iv) exhibited rearranged C delta and J delta and its mRNA. An NK-like activity and IL-2 production were induced in the TCR beta delta and gamma delta cell lines [group (ii) and (iii)] by treatment with PHA and PMA.

Antibodies, Monoclonal↗
PubMed1989

Formation of multinucleated cells in a Hodgkin's-disease-derived cell line.

The proliferative potential of multinucleated Reed-Sternberg-like cells and the process of multinuclear formation were studied on the Hodgkin's-disease-derived cell line HDLM-2. No difference in surface antigen expression was found between mono- and multinucleated cells as determined by immunolabelling with characteristic markers. After sorting and reculture of purified mononucleated cells, polykaryons emerged subsequently in these cultures, indicating that mononucleated cells give rise to multinucleated variants. The morphological observation of mitotic figures and immunostaining with the cell cycle indicators Ki-67 and BrdU provided evidence of DNA synthesis and nuclear division in multinucleated cells. The presence of mitotic figures demonstrated that multinucleated cells are able to undergo synchronous nuclear division. However, while polykaryons were clearly mitotically active and capable of DNA synthesis, the absence of telophases and the failure of active replication suggest a disturbed cytokinesis. Co-cultivation of BrdU-labelled and unlabelled populations did not lead to hybrid polykaryons with negative and positive nuclei. Therefore, multinucleated giant cell formation of HDLM-2 cells appears to involve nuclear endomitosis without cell division rather than cell fusion.

Antibodies, Monoclonal↗
PubMed1989

Expression of protein kinase C subspecies in human leukemia-lymphoma cell lines.

Expression of protein kinase C (PKC) subspecies was studied in various human leukemia-lymphoma cell lines. The PKC in most cell lines examined was resolved into two major fractions corresponding to type II (beta-sequence) and type III (alpha-sequence) PKC of the rat brain. The amounts of these two subspecies greatly varied among the cell lines. Type I PKC (gamma-sequence) was expressed in none of the cell lines tested, but PKCs with undefined structures were frequently detected. The differential co-expression of several PKC subspecies is presumably related to the state of cell differentiation.

Animals↗
PubMed1989

Imbalance of deoxyribonucleoside triphosphates, DNA double-strand breaks, and cell death caused by 2-chlorodeoxyadenosine in mouse FM3A cells.

The mechanism of the cytotoxic action of 2-chlorodeoxyadenosine in mouse FM3A cells was investigated. Imbalance of the dNTP pools occurred within 3 h of treatment with 20 microM 2-chlorodeoxyadenosine; the dATP and dGTP pools were depleted and the dTTP pool increased. 2-Chlorodeoxyadenosine added to the culture medium broke mature DNA strands, giving fragments of 100-200 kilobase pairs as found by orthogonal-field-alternation gel electrophoresis. DNA strand breaks, measured by this technique, were observed in the treated cells about 12 h after the addition. The cells also lost viability at about 12 h. Breaks in the single and double strands of DNA, as measured by alkaline and neutral filter elution, became evident 18 h after treatment with 20 microM 2-chlorodeoxyadenosine; there were as many single-strand breaks as would be caused by 130 rads of gamma-ray irradiation. Double-strand breaks were equivalent to those caused by 2180 rads of gamma-ray irradiation. Comparison of the ratio of single- and double-strand breaks caused by 2-chlorodeoxyadenosine to that following radiation suggested that 2-chlorodeoxyadenosine broke only double strands. Cycloheximide inhibited the breakage of DNA double strands and the cell death caused by this compound. Flow cytometric studies of cytostasis brought about by 2-chlorodeoxyadenosine in FM3A cells showed that cells accumulated in the earlier part of the S phase. 2-Chlorodeoxyadenosine decreased DNA synthesis more than RNA or protein synthesis. The breaks in double strands of DNA were probably important in the cell death caused by 2-chlorodeoxyadenosine. The intracellular dNTP imbalance may trigger these events.

Animals↗
PubMed1989

Characterization of Hodgkin's disease derived cell line HDLM-2.

The cell line HDLM-2 was established from the pleural effusion of a patient with Hodgkin's disease. Here, we describe the morphological, cytochemical, enzymological, immunological, molecular biological, and functional characteristics of the cell line. The results of this multiparameter profile show that HDLM-2 is different from other well-studied leukemia-lymphoma cell lines including other Hodgkin's disease derived cell lines. HDLM-2 cultures contain mainly mono- or binucleated cells, but also prominent giant cells with two to ten nuclei. HDLM-2 cells do not express an immunophenotype characteristic of a given cell lineage. However, the cells are positive for Ki-1, HeFi-1, Leu-M1, Tac, and HLA class II markers. Cytochemical, enzymological, and functional data are equally inconclusive, but are definitely not compatible with a monocyte/macrophage profile. Analysis of the gene status documents that T-cell receptor beta- and gamma-chain genes are rearranged while immunoglobulin heavy chain genes are in germline configuration. The combined results indicate a T-cell origin of HDLM-2 cells. The evidence available from this and other established Hodgkin's disease derived cell lines suggests a lymphoid origin of Hodgkin and Reed-Sternberg cells.

Animals↗
PubMed1989

T-cell receptor gene rearrangement and its expression in human myeloid leukemia cell lines.

ML cell lines (ML-1, -2, and -3) were derived from the cells of a patient with acute myelocytic leukemia preceded by a T-cell malignant lymphoma. A deletion of chromosome 11 (11q-) was common to the affected cells in both neoplastic phases. We report here that the three ML cell lines have DNA rearrangements of the T-cell receptor (TcR)-beta and gamma-chain genes in addition to immunoglobulin heavy-chain gene rearrangement, though they do not have TcR gene messages. The findings presented here indicate that ML cell lines could be used as models for the elucidation of the bilineal nature of hematopoietic neoplastic cells, though they have a biphenotypic (myelomonocytic/T-cell) marker expression.

Blotting, Northern↗
PubMed1989

Establishment and characterization of a clonal human T-cell line, MKB-1 derived from a patient with acute myeloblastic leukemia.

A human T-cell line, designated as MKB-1, was established by cloning procedures in a suspension culture from a peripheral blood of a 17-year-old female patient with acute myeloblastic leukemia. The immunological marker profile of MKB-1 indicated that unlike a myeloid phenotype of the original leukemic cells, the cells were positive for CD3 (both cell surface and cytoplasm), T cell receptor (TcR) alpha/beta heterodimer, CD4, CD5, CD7, CD10, CD57 (Leu7), SN-1 and the cytoplasmic TcR beta chain. These findings indicate the T cell nature of the established cells. Terminal deoxynucleotidyl transferase (TdT) was also detected in 60%. We did not detect markers of human myeloid and B cell associated antigens, HLA-class II or immunoglobulin chains. Cytogenetic study revealed that the MKB-1 cells had a female hypo-tetraploid karyotype with chromosomal abnormalities including a translocation between chromosomes 10 and 14. The breakpoint of chromosome 14 of this translocation, 14q11.2, is known to be the location of TcR alpha and delta genes; t(10; 14) (q26; q11.2) is a variant type of a T cell neoplasm-associated translocation, t (10; 14) (q24; q11.2). The MKB-1 cell line is unusual in that its T cell characteristics are phenotypically and cytogenetically distinct from the original myeloid leukemia cells.

Adolescent↗
PubMed1989

Two pathways for inducing interleukin 2 production in MOLT 16 cells, a human leukemic T-cell line: interleukin 1 pathway and interleukin 1-independent pathway.

Phytohemagglutinin (PHA)-stimulated MOLT 16 cells (a human leukemic T cell line, at a concentration of 2 x 10(6) cells/ml) secreted 6 U/ml of interleukin 2 (IL 2) into the culture supernatant during 24 h culture. When interleukin 1 (IL 1) (5 U/ml) or IL 1-producing cells such as human T cell leukemia virus-1 (HTLV-1)-transformed T cell line, C5/MJ, and myelomonocytic cell line, THP-1-O, were added to the MOLT 16 culture at a concentration of 4 x 10(5) cells/ml, far greater IL 2 production (greater than 65 U/ml) was observed. The activity of soluble IL 1 and membrane-associated IL 1 produced by these accessory (A) cells was completely neutralized by the treatment with anti-human IL 1 antibody. In sharp contrast, MOLT 16 cells co-cultured with BALL-1 cells (a human leukemic B cell line) resulted in comparable increases in IL 2 production (175 U/ml), although no IL 1 or IL 1-like activity was detected either in the supernatants or in the cell lysates of BALL-1 cells. An augmentation of IL 2 production was also induced with paraformaldehyde-fixed BALL-1 cells, and this augmentation could not be inhibited by anti-human IL 1 antibody. These data indicate that MOLT 16 provides a model for determining pathways for activating T cells leading to IL 2 production in response to mitogens. Two pathways have been described in the report: one involving secreted or membrane-associated IL 1, and a second independent pathway which involves contact of surface membranes in the presence of PHA.

Antigens, Surface↗
PubMed1988

myb oncogene in human hematopoietic neoplasia with 6q- anomaly.

Molecular and cytogenetic analyses were performed on human T-cell leukemia cell lines (PEER and MOLT-4) with the 6q- anomaly. The PEER cells contained an interstitial deletion of the long arm of chromosome 6, that is, del(6)(q13q21), as well as other changes. The MOLT-4 cells showed a terminal deletion of the long arm of chromosome 6, that is, del(6)(q24). The 700-bp BamHI/XbaI-digested c-myb probe hybridized to a 4.3-kb fragment in EcoRI digested DNAs from these two cell lines, showing no deletion, rearrangement, or amplification. On the other hand, ML cells [ML-1, -2 and -3; human myeloid/T-cell biphenotypic leukemia cell lines with del(6)(q24)] showed an amplification of the c-myb gene and had a high level of the c-myb-related mRNA at 3.5 kb. Though no amplification of the c-myb at the DNA level was noted in the PEER or MOLT-4 cell lines, apparent high expression of the c-myb was detected in these human T-cell neoplastic lines. These results indicate that high c-myb expression is related to lineage of hematopoietic neoplasia rather than to the 6q- change.

Chromosome Banding↗