Biomedical subjects

N Nara

Publications and source records attributed to N Nara.

At least 73 records · Page 4Linked to original sources

PubMed1990

Effects of recombinant human granulocyte colony-stimulating factor on the hematologic recovery and survival of irradiated mice.

We studied the effects of intraperitoneal injections of recombinant human granulocyte colony-stimulating factor (rhG-CSF) according to various administration schedules on the recovery of spleen colony-forming units (CFU-S) and peripheral blood counts, and on the survival of irradiated mice. The sooner and more frequently the mice were injected with rhG-CSF after irradiation, the more enhanced the recovery of CFU-S in bone marrow was obtained on day 7. Twice-daily injections of rhG-CSF from day 0 to day 2 significantly enhanced the recovery of platelets and hematocrit, but two injections of rhG-CSF on only day 0 did not. Twice-daily injections of rhG-CSF from day 0 to day 6 enhanced the recovery of platelets more effectively than twice-daily injections of rhG-CSF from day 1 to day 7, and increased the survival of irradiated mice more effectively than any other examined administration schedules. Twice-daily injections of rhG-CSF from day 0 to day 6 were significantly effective in enhancing the survival of mice irradiated with 8.5-, 9.0-, and 9.5-Gy x-rays, although not effective after irradiation of 10.5-Gy x-rays.

Animals↗

PubMed1990

Richter syndrome with two B cell clones possessing different surface immunoglobulins and immunoglobulin gene rearrangements.

A patient with two populations of B cell malignancy in the bone marrow is reported. One population consisted of mature small lymphocytes, expressing surface IgM + D, lambda and proliferating very slowly. The other population consisted of abnormal large lymphoid cells, expressing surface IgM, K and proliferating actively. We considered the former as chronic lymphocytic leukemia cells and the latter as malignant lymphoma cells. Therefore, this case was considered as Richter syndrome. The analysis of immunoglobulin heavy chain gene rearrangement showed the different patterns between the two populations. It suggested that the two populations arose from the different origins. We discussed the genetic relationship between the two clones of Richter syndrome referring to other reported cases.

Aged↗

PubMed1990

Effects of interleukin-6 and granulocyte colony-stimulating factor on the proliferation of leukemic blast progenitors from acute myeloblastic leukemia patients.

The effects of recombinant human interleukin-6 (rh IL-6), which has homology with rh granulocyte colony-stimulating factor (rh G-CSF) at the amino acid sequence level, and rh G-CSF on normal human bone marrow cells, fresh leukemic blast progenitors from 16 acute myeloblastic leukemia (AML) patients, and G-CSF-dependent human AML cell line (OCI/AML 1a) were investigated. rh G-CSF stimulated the proliferation of leukemic blast progenitors from 13 out of 16 AML patients tested. rh IL-6 stimulated the proliferation of blasts from eight AML patients and enhanced the G-CSF-dependent proliferation of the fresh AML blasts from two out of eight patients tested. On the other hand, rh IL-6 suppressed the blast colony formation from two AML patients and OCI/AML 1a cells and also reduced the G-CSF-dependent proliferation of the blast progenitors from one of the two patients and the cell line, rh IL-6 had no effect on the colony formation of normal granulocyte-macrophage colony-forming units (CFU-GM) with or without rh G-CSF. Differentiation-induction by rh IL-6 was not observed in the fresh AML blasts but was observed in OCI/AML 1a. The effect of IL-6 on the blast colony formation and G-CSF-dependent blast cell growth was complicated and heterogenous among the AML cases; IL-6 stimulated blast colony formation in some cases and suppressed it in others. The heterogeneity of the response was supposed to be derived from the heterogeneity of the characteristics of AML cells. Although G-CSF simply stimulated the blast colony formation, IL-6 had a bimodulatory effect on the proliferation of leukemic blast progenitors from AML patients. IL-6 might be involved in the regulation of the proliferation of AML cells in vivo as well as in vitro.

Adolescent↗

PubMed1990

Comparative effects of busulfan, cytosine arabinoside and adriamycin on different maturation stages of normal human bone marrow cells.

Leukemic blast progenitors in acute myeloblastic leukemia (AML) undergo terminal divisions and/or self-renewal, which can be studied by the methylcellulose culture method and suspension culture, respectively. Using these methods, we have shown that busulfan (BU) was more effective against the self-renewal of blast progenitors than against the terminal divisions. As reported previously, cytosine arabinoside (Ara-C) is more effective against self-renewal and adriamycin (ADR) is more effective against terminal divisions. To determine the reason why antileukemic drugs show different sensitivity against self-renewal and terminal divisions, we studied the effects of these drugs on normal human bone marrow cells. We have shown that BU and Ara-C suppressed the colony formation induced by interleukin-3 (IL-3) more effectively than that by granulocyte colony-stimulating factor (G-CSF). On the other hand, ADR suppressed the colonies induced by G-CSF more effectively. In normal hemopoiesis, IL-3 has been shown to stimulate the growth of more primitive hemopoietic stem cells, while G-CSF has been documented to stimulate the growth of granulopoietic committed precursors at a more differentiated stage. Therefore, BU and Ara-C are considered to be more effective against the immature cells in normal bone marrow and ADR is effective against the more differentiated cells. We suppose that the different differentiation stage of target cells may explain the difference of the effects on self-renewal and terminal divisions between BU, Ara-C and ADR.

Bone Marrow↗

PubMed1990

Effect of platelet-derived growth factor on the proliferation of blast progenitors from acute myeloblastic leukemia patients.

The effect of platelet-derived growth factor (PDGF) on self-renewal and terminal divisions of blast progenitors of acute myeloblastic leukemia was studied. The terminal divisions of blast progenitors were determined as the primary blast colony formation in methylcellulose culture; self-renewal was assessed by secondary colony formation by replating in methylcellulose and the recovery of clonogenic cells in suspension culture. PDGF neither enhanced nor suppressed primary or secondary blast colony formation in methylcellulose culture nor recovery of clonogenic cells in suspension. The results show that PDGF does not affect the self-renewal or terminal divisions of leukemic blast progenitors.

Adolescent↗

PubMed1990

Role of humoral and cellular factors on the growth of blast progenitors of acute myeloblastic leukemia in serum-free culture.

To determine the growth requirement of leukemic blast progenitors in acute myeloblastic leukemia (AML), leukemic cells from the peripheral blood of eight AML patients were cultured in the serum-free culture system. Blast progenitors made colonies in methylcellulose culture and showed exponential growth in suspension culture, although the growth of blast progenitors in the absence of fetal calf serum (FCS) in some patients was inferior to that in the FCS-enhanced culture system. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) stimulated the growth of blast progenitors in a dose-responsive manner. When cells were cultured at high cell density, blast colonies were formed even in the absence of CSF. Irradiated blasts also supported the growth of intact blast progenitors. These results confirm the finding noted in the FCS-enhanced culture studies that granulopoietic factor, G-CSF, plays an important role on the leukemic growth. The importance of cell to cell interaction for the growth of blast progenitors was also confirmed.

Blast Crisis↗

PubMed1990

Terminal differentiation to mature neutrophils and eosinophils in suspension culture of the blast progenitors in acute myeloblastic leukemia.

The blasts obtained from three freshly diagnosed acute myeloblastic leukemia (AML) patients were cultured in suspension to determine whether leukemic blast progenitors can indeed differentiate to form mature granulocytes. One patient was AML M2. The other two patients were bilineal and biphenotypic leukemia, respectively. Media conditioned by human bladder carcinoma line 5637 (5637-CM) or recombinant human granulocyte colony-stimulating factor (rhG-CSF) was added to stimulate growth. In suspension, clonogenic cells grew for 1-3 weeks in two patients, while they did not increase in one patient. After repeated subculture, cells of blast morphology decreased in percentage and polymorphonuclear neutrophils, eosinophils, and monocyte-macrophages appeared. Lymphoid cell component of the patient 2, who was diagnosed as bilineal leukemia by dual-color immunofluorescence analysis, decreased in number after suspension culture and cells of myeloid phenotype became dominant. The findings show that clonogenic blast progenitors can renew themselves and can also undergo terminal differentiation to mature end cells.

Adult↗

PubMed1990

Haematopoietic suppressing activity of gamma-interferon in serum and bone marrow of aplastic anaemia patients.

To determine the mechanism by which haematopoiesis is suppressed in aplastic anaemia, the effect of the sera from 6 patients on the granulopoietic precursors (colony-forming units in culture; CFU-C) was studied in vitro. Addition of the sera from 2 patients significantly suppressed CFU-C. The suppressive effect of the sera on CFU-C was inhibited by the addition of 1.5 NU/ml of anti-gamma-IFN antibody. In another patient, anti-gamma-IFN antibody increased autologous CFU-C although the serum of the patient did not suppress CFU-C. Serum gamma-IFN levels of all patients were under 2 IU/ml. The above findings suggest that humoral factors inhibit haematopoiesis in some patients with aplastic anaemia, and that gamma-IFN plays a role as an inhibitor even at a low concentration.

Adolescent↗

PubMed1990

Mechanism of action of interleukin 1 on the progenitors of blast cells in acute myeloblastic leukemia.

We tested the effect of interleukin 1 (IL-1) on the growth of leukemic blast progenitors from patients with acute myeloblastic leukemia (AML). A purified blast cell fraction depleted of both T cells and phagocytic cells was tested at different cell densities. Addition of 1 ng/ml of IL-1 alpha alone enhanced blast colony formation in 10 of 13 cases tested, and the enhancement was prominent when plated cell densities were lowered. The conditioned media (CM) from AML patients contained varied levels of IL-1 activity, and following depletion of phagocytic cells, the levels decreased markedly in all cases tested. Addition of either antiserum against IL-1 alpha or IL-1 beta reduced the IL-1 activity in CM, suggesting that AML blasts produce both IL-1 alpha and IL-1 beta. Addition of IL-1 alpha or IL-1 beta antiserum inhibited blast colony formation in a dose-dependent manner, and a combination of both antisera showed the most marked inhibition. However, the augmentation of blast colony formation was almost completely inhibited by addition of anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) serum in all three cases tested. IL-1 is also devoid of this activity when tested in the presence of a combination of granulocyte CSF (G-CSF), GM-CSF, and interleukin 3 (IL-3) at an optimal concentration. These results suggest that blast cells could produce and secrete CSF(s) and/or IL-1, and that the growth-enhancing effect of IL-1 on AML blasts is indirect, via production of CSFs by leukemic cells.

Colony-Stimulating Factors↗

PubMed1989

Antiproliferative and differentiative effect of granulocyte-macrophage colony-stimulating factor on a variant human small cell lung cancer cell line.

A variant clone was adopted during passages of a small cell lung cancer cell line, GKT3-1.3. The variant clone exhibited distinct characteristics with alterations in morphology, positive staining with nonspecific esterase stain, and an increase in surface specific markers OKM5, HLA-DR, Mo1, and My7, usually found on monocytes or their precursors. However, it exerted a very rapid proliferation just like immature cells. This new clone, GKT3-1.3V, was shown to have specific binding capacity to granulocyte-macrophage colony-stimulating factor (GM-CSF), with a number of binding sites comparable to that of myelomonocytes or monocytic cell lines. Thus its proliferation was inhibited by GM-CSF in clonogenic assay and suspension culture. Increase in the percentage of cells with surface marker Mo1 by the addition of GM-CSF suggested its differentiative effect. Cell cycle analysis showed that the antiproliferative effect of GM-CSF was due to a block in G0 or G1. The antiproliferative effect of GM-CSF was abolished by the addition of anti-GM-CSF antibody.

Carcinoma, Small Cell↗

PubMed1989

Expression of interleukin 1 beta receptors on blast cells in acute myeloblastic leukemia: comparison with interleukin 1 beta proliferative activity.

We describe here the presence of a single class of interleukin 1 beta (IL-1 beta) receptors on the surface of blast cells freshly obtained from eight acute myeloblastic leukemia patients and one chronic myelocytic leukemia patient in blast crisis. Blast cells possessed a low number of high-affinity receptors (range, less than 10-173 receptors/cell) with a Kd of 1.8-12.8 x 10(-10) M. At the same time, we have investigated the effects of IL-1 on the growth of leukemic blast progenitors, and a significant heterogeneity of responsiveness was observed. IL-1 beta (1 ng/ml) enhanced blast colony formation in six patients. No significant effect was observed by addition of up to 100 ng/ml of IL-1 beta in the remaining three patients. No significant correlation was observed between the receptor number, receptor affinity, and the cellular responsiveness to IL-1; in some acute myeloblastic leukemia cases with apparent IL-1 receptors, no proliferation response to added IL-1 was observed. Our data show that IL-1 alone can enhance blast colony formation and that lack of responsiveness to IL-1 in some acute myeloblastic leukemia patients is not related to the absence of IL-1 receptors on blast cells.

Adult↗

PubMed1989

Autoantibody against platelet glycoprotein IIb/IIIa in a patient with non-Hodgkin's lymphoma.

An autoantibody to platelet glycoprotein (GP) II b/III a was produced in a 38 year-old woman who had a previous history of the malignant lymphoma of the stomach. The aggregations of the patient's platelets showed losses of the primary waves in response to ADP and epinephrine and marked hypoaggregation in response to collagen, while agglutination by ristocetin was normal. Crossed immuno-electrophoresis (CIE) of her platelets solubilized by 1% Triton X-100 revealed an abnormal biphasic precipitate line of GP II b/III a complex. Nine months later, she developed severe thrombocytopenia along with a relapse of the lymphoma in the cervical lymph nodes. The patient's IgG, which was collected during her thrombocytopenic period and purified, inhibited ADP-, epinephrine- and collagen-induced aggregations of normal platelets. In CIE, the 125I-labelled IgG of the patient, inserted into the intermediate gel, was incorporated into the precipitation line of the GP II b/III a complex of normal platelets. Radiation treatment to the cervical lymph nodes dramatically normalized both the function and the count of the patient's platelets. From these findings, it is suggested that an autoantibody to the GP II b and/or III a was produced by the lymphoma cells.

Adult↗

PubMed1989

Specific binding of radioiodinated human GM-CSF to the blast cells of acute myeloblastic leukemia.

We describe here the presence of two classes of binding sites for GM-CSF expressed on blasts freshly isolated from five AML patients and one patient with CML in blastic phase: one of high-affinity (38-177 per cell, KD 8-150 pM) and one of low-affinity (121-806 per cell, KD 503-2683 pM). No correlation is observed between the receptor number, receptor affinity, and the growth stimulatory effect of GM-CSF on leukemic blast progenitors. Blasts from two cases showed no or negligible response to GM-CSF but expressed comparable numbers of receptors when compared with the numbers expressed by the sensitive blasts. Our data suggest that significant proliferative effects of GM-CSF can occur at low levels of high-affinity receptor occupancy. Lack of responsiveness to GM-CSF in some AML patients is not correlated to the absence of GM-CSF receptors on leukemic cells. Reduction in the growth of blast progenitors at high concentrations of GM-CSF may be attributed to the differentiating activity of GM-CSF via low-affinity receptors on leukemic cells.

Binding, Competitive↗

PubMed1989

Binding properties and proliferative effects of human recombinant granulocyte-macrophage colony-stimulating factor in primary leukemia and lymphoma.

Binding of radiolabeled human granulocyte-macrophage colony-stimulating factor (GM-CSF) was studied with blast cells from eight patients with acute myeloblastic leukemia (AML), and neoplastic lymphoid cells from one patient with acute lymphoblastic leukemia (ALL), two patients with chronic lymphocytic leukemia (CLL) and one patient with undiagnosed B cell neoplasia. In all AML cases studied, Scatchard graphs of the direct binding data were curvilinear, and were best fitted by curves derived from a two-binding-site model; one site with high affinity (Kd1 = 12-71 pM; 174-602 sites/cell) and the other with low affinity (Kd2 = 0.5-2.7 nM; 1137-6020 sites/cell). A cross-linking study on blast cells from one AML patient demonstrated specific bands which were similar to those reported for peripheral blood neutrophils. Furthermore, blast colony assays for the same preparations showed remarkable proliferative response to GM-CSF in the concentration range from 0.3 nM to 7.0 nM (ED50 greater than 0.7 nM). This concentration range is approximately one order of magnitude higher than that which is effective for colony formation from normal bone marrow progenitors (ED50 in equilibrium 0.1 nM). No significant correlation could be observed between the responsiveness of blast progenitors to GM-CSF, and the numbers or affinities of GM-CSF binding sites demonstrated on blast cells. In studies with neoplastic lymphoid cells from four patients, 125I-GM-CSF also specifically bound in two cases, while response to GM-CSF was not observed in these cases. These results indicate that the expression of GM-CSF receptor is not restricted to the GM-CSF-responsive AML blast cells, but can be observed in other AML blast cells and even in neoplastic lymphoid cells.

Binding Sites↗

PubMed1989

The effects of vincristine and doxorubicin on the clonogenic cells of a human lung cancer cell line in methylcellulose and suspension culture.

The effects of vincristine (VCR) and doxorubicin (DOX) on the growth of an established line of human lung cancer cells, PC9, were studied in methylcellulose and suspension cultures. The secondary colony formation in methylcellulose and recovery of clonogenic cells in suspension were considered to reflect well the self-renewal of the clonogenic cells. When dose-response curves were obtained for VCR and DOX, the primary clonogenic cells (PE1) were more sensitive than secondary clonogenic cells (PE2) or clonogenic cells in suspension. Repeated exposure to VCR in suspension did not inhibit the exponential growth of the clonogenic cells. These data indicated that both drugs were relatively ineffective in specifically suppressing the self-renewal of the clonogenic cells.

Cell Division↗

PubMed1989

Effects of verapamil on the cellular accumulation of daunorubicin in blast cells and on the chemosensitivity of leukaemic blast progenitors in acute myelogenous leukaemia.

Verapamil, a calcium channel blocker, was studied for its effects on the cellular daunorubicin (DNR) accumulation in blast cells and on the sensitivity of the blast progenitors to DNR in 30 acute myelogenous leukaemia (AML) patients. Using flow cytometry, verapamil was shown to increase the accumulation of DNR in blast cells. The effect was more prominent in the patients who showed poorer response to chemotherapy including DNR. The per cent increases of DNR content by verapamil were 6.4 +/- 6.3% and 19.5 +/- 23.1% in the 16 responders and the 14 nonresponders, respectively (P less than 0.05). The data suggest the presence of enhanced efflux of DNR in nonresponders. Marked variation in the effects of verapamil among nonresponders suggests the heterogeneity of the mechanisms of drug resistance involved. Verapamil also enhanced the sensitivity of blast progenitors to DNR. The degree of increase of cellular DNR accumulation by verapamil correlated with the degree of increase in chemosensitivity of blast progenitors (nonresponders, P less than 0.005; responders, P less than 0.05). We conclude that enhanced efflux of DNR in blast progenitors may be related to remission induction failure in at least some of resistant AML patients.

Adult↗

PubMed1989

Inhibition of the in vitro growth of blast progenitors from acute myeloblastic leukemia patients by transforming growth factor-beta (TGF-beta).

The effects of transforming growth factor-beta (TGF-beta) on the blast progenitors from nine acute myeloblastic leukemia patients were studied in methylcellulose and suspension cultures. Leukemic blast progenitors undergo terminal divisions with a limited differentiation in methylcellulose culture, making blast colonies. Blast progenitors can renew themselves. The self-renewal can be reflected by secondary colony formation after replating primary blast colonies in fresh methylcellulose media and by the exponential growth of clonogenic cells in suspension culture. TGF-beta suppressed primary and secondary colony-formation in methylcellulose culture. Furthermore, TGF-beta suppressed the recovery of clonogenic cells in suspension. The results indicate that TGF-beta is effective in inhibiting not only terminal divisions but also self-renewal of leukemic blast progenitors.

Adolescent↗

PubMed1989

Autocrine growth mechanisms of the progenitors of blast cells in acute myeloblastic leukemia.

Autocrine growth mechanisms of leukemic blast progenitors in acute myeloblastic leukemia (AML) were investigated. Colony formation of leukemic blast progenitors was observed in 14 of 14 patients tested when purified blast cell fraction depleted of both T cells and monocytes was plated in methylcellulose without any colony-stimulating factor (CSF). However, there existed a minimal cell density required to initiate blast progenitor growth with marked patient-to-patient variation. To clarify the role of cell density on the spontaneous growth of blast progenitors, we tested whether leukemic cells produced and secreted some stimulatory humoral factor(s). Production of colony-stimulating activity (CSA) by blast cells was observed in 17 of 18 patients tested. Following further depletion of monocytes, the CSA levels decreased markedly in 14 patients, indicating that blast cells with monocytoid differentiation were responsible for CSA production. We also confirmed granulocyte colony-stimulating factor (G-CSF) and/or granulocyte macrophage-colony-stimulating factor (GM-CSF) production by leukemic blasts using specific immunologic assays. When leukemic cells were divided into nonadherent nonphagocytic cell fraction and adherent cell fraction, only nonadherent nonphagocytic cells showed clonogenecity and adherent blast cells lacked the colony-forming capacity. The results indicate that there are at least two blast cell subpopulations in AML: one is proliferating subpopulation with self-renewal capacity and the other is supporting subpopulation with functions such as CSF production. The quite intimate relationship between these two blast cell subpopulations in AML may play an important role on the growth of leukemic blast progenitors in vitro.

Adult↗