losartan-potassium and Leukemia--Myeloid

Topics: Acute Disease; Aged; Azacitidine; Chelation Therapy; Chromosome Deletion; Chromosomes, Human, Pair 5; Clinical Trials as Topic; Decitabine; Disease Progression; DNA Methylation; Erythropoietin; Hematopoietic Cell Growth Factors; Hematopoietic Stem Cell Transplantation; Humans; Iron Chelating Agents; Lenalidomide; Leukemia, Myeloid; Myelodysplastic Syndromes; Severity of Illness Index; Thalidomide; Transplantation, Homologous

Bone marrow suppression after intensive chemotherapies in patients with myeloid leukemia is severe, resulting in the reduction of the number of white blood cells, red blood cells, and platelets. Supportive therapies are indispensable for the management of these leukemia patients. The improvement of blood cell transfusion can decrease side effects of chemotherapies and establish the safety. But we still have notable side effects of transfusion such as TRALI (transfusion-related acute lung injury), platelet immunologic refractory state, and so on. Cytokine therapy especially with G-CSF (granulocyte colony-stimulating factor) administration, changed the treatment of myeloid leukemia. G-CSF can shorten the duration of neutropenia and decrease the risk of infection. Recently the effects of Epo (erythropoietin) on chemotherapy-induced anemia have been demonstrated. We discuss here the indications of blood cell transfusion and cytokine therapies in the treatment for myeloid leukemia.

Topics: Acute Lung Injury; Antineoplastic Agents; Blood Component Transfusion; Blood Transfusion; Erythropoietin; Graft vs Host Disease; Granulocyte Colony-Stimulating Factor; Humans; Leukemia, Myeloid; Practice Guidelines as Topic; Transfusion Reaction

Topics: Anemia; Apoptosis; Clone Cells; Disease Progression; Erythropoiesis; Erythropoietin; Female; Growth Substances; Hematologic Tests; Hematopoietic Stem Cells; Humans; Iron; Leukemia, Myeloid; Male; Myeloproliferative Disorders; Phlebotomy; Phosphorus Radioisotopes; Polycythemia Vera; Pregnancy; Pregnancy Complications, Hematologic; Receptors, Erythropoietin; Receptors, Growth Factor; Thrombocytosis; Thrombosis

With the identification of recombinant production of the hematopoietic growth factors, these cytokines have been evaluated in the treatment of primary bone marrow failure states and after myelosuppressive chemo- or radiotherapy. Granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and erythropoietin have been approved for clinical use, and others including c-mpl-ligand (also called megakaryocyte growth and development factor or thrombopoietin) are in phase I and II trials. Most studies have been done with granulocyte and granulocyte-macrophage colony-stimulating factors; their beneficial effects are proven regarding acceleration of granulocyte recovery after chemo- and radiotherapy. In the majority of trials, this acceleration results in a reduction of infectious risks, a shortening of drug- and radiation-induced myelosuppression, and a higher chemotherapy dose intensity; however, an improved remission rate and improved long-term survival rates have not yet been definitively documented. Guidelines have been published to provide a rational basis for the use of these factors in clinical practice. It should be emphasized, however, that for many of the recommendations data from randomized clinical trials are lacking.

Topics: Acute Disease; Antineoplastic Agents; Clinical Trials as Topic; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Stem Cell Transplantation; Humans; Leukemia, Myeloid; Neoplasms; Recombinant Proteins; Thrombopoietin

Hematopoiesis is regulated in a very precise fashion by a balance between both positive and negative growth factor signals in the hematopoietic microenvironment. Many of these growth factor signals have been identified and are available as recombinant proteins. At the present time, the approved uses for hematopoietic growth factors involve their use to facilitate blood cell production in situations of hematopoietic suppression or in cases where endogenous growth factor production is inappropriately low. Recent studies with the hematopoietic growth factors are looking at innovative strategies for their use and new clinical situations in which to evaluate their effectiveness. This review looks at some of these new uses for hematopoietic growth factors.

Topics: Animals; Bone Marrow Diseases; Bone Marrow Transplantation; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Leukemia, Myeloid

Topics: Animals; Colony-Stimulating Factors; Erythropoietin; Growth Substances; Hematopoiesis; Hematopoietic Cell Growth Factors; Humans; Leukemia, Myeloid; Leukopenia; Mice; Myelodysplastic Syndromes; Recombinant Proteins

Topics: Animals; Bone Marrow Cells; Cell Differentiation; Cell Division; Cells, Cultured; Colony-Stimulating Factors; Dose-Response Relationship, Drug; Erythroblasts; Erythrocytes; Erythropoiesis; Erythropoietin; Hematopoiesis; Hematopoietic Stem Cells; Humans; Iron Radioisotopes; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Mice; Polycythemia

Topics: Cell Differentiation; Cell Division; Cells, Cultured; Colony-Forming Units Assay; Colony-Stimulating Factors; Erythroblasts; Erythropoiesis; Erythropoietin; Growth Substances; Hematopoietic Stem Cells; Hormones; Humans; Interleukin-3; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Lymphokines; Neoplastic Stem Cells; Polycythemia Vera; Primary Myelofibrosis; Thrombocytosis

Topics: Animals; beta 2-Microglobulin; Blood Cells; Cell Differentiation; Cell Division; Cell Membrane; Cells, Cultured; Colony-Stimulating Factors; Culture Media; Dialysis; Erythrocytes; Erythropoietin; Glycoproteins; Granulocytes; Hematopoietic Stem Cells; Histocompatibility Antigens; Humans; Lectins; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Thymidine

We report long-term results of treatment of myelodysplastic syndrome (MDS) with erythropoietin and granulocyte colony-stimulating factor (G-CSF). A total of 129 patients were followed up 45 months after last inclusion in the Nordic MDS Group studies. Erythroid response rate was 39% and median response duration 23 months (range, 3-116 months or more). Complete responders showed longer response duration than partial responders (29 versus 12 months, P = .006). The International Prognostic Scoring System (IPSS) groups Low/Intermediate-1 (Low/Int-1) had longer response duration than Int-2/High (25 versus 7 months, P = .002). The time until 25% developed acute myeloid leukemia (AML) was longer in the good and intermediate predictive groups for erythroid response compared with the poor predictive group (52 versus 13 months, P = .008). Only 1 of 20 long-term responders developed AML. We assessed the effect on long-term outcome by comparing treated patients with untreated patients selected from the IPSS database using multivariate Cox regression, adjusting for major prognostic variables. There was no difference in survival (odds ratio [OR], 0.9; 95% confidence interval [CI], 0.7-1.2; P = .55) or risk of AML evolution (OR, 1.3; 95% CI, 0.7-2.2; P = .40) between treated and untreated patients. Patients with high/intermediate probability of response and with IPSS Low/Int-1 show frequent and durable responses without adverse effects on outcome, while other patients should not be considered candidates for this treatment.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Analysis of Variance; Anemia; Cell Transformation, Neoplastic; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Humans; Leukemia, Myeloid; Male; Middle Aged; Myelodysplastic Syndromes; Predictive Value of Tests; Prognosis; Survival Analysis; Treatment Outcome

Pluripotent stem cells of hematopoiesis and lymphopoiesis are among the CD34+ cells in blood or bone marrow. After granulocyte-colony stimulating factor (G-CSF) treatment, 1% to 2% of the mononuclear cells in blood are CD34+ cells, which can be procured by leukapheresis. We investigated the potential of CD34+ blood cells for reconstituting hematopoiesis and lymphopoiesis after allogeneic transplantation. HLA-identical sibling donors of 10 patients with hematologic malignancies were treated with G-CSF (filgrastim), 5 microgram/kg subcutaneously twice daily for 5 to 7 days. CD34+ cells were selected from the apheresis concentrates by immunoadsorption, concomitantly the number of T cells was reduced 100- to 1,000-fold. After transplantation, five patients received cyclosporine A for graft-versus-host disease (GvHD) prophylaxis (group I); five patients additionally received methotrexate (group II). G-CSF and erythropoietin were given to all patients. Mean numbers of 7.45 x 10(6) CD34+ and 1.2 x 10(6) CD3+ cells per kilogram were transplanted. In group I, the median times of neutrophil recovery to 100, 500, and 1,000 per mm3 were 10, 10, and 11 days, respectively. Group II patients reached these neutrophil levels after 10, 14, and 15 days, respectively. Platelet transfusions were administered for a median of 18 days in group I and 30 days in group II, and red blood cells for 9 and 12 days, respectively. Between day 30 and 60, lymphocytes reached levels of 353 +/- 269 cells per mm3. The median grades of acute GvHD were III in group I and I in group II. Two patients in group I died from acute GvHD. Two leukemic relapses occurred in group II. Complete and stable donor hematopoiesis was shown in all patients with a median follow up of 370 (45 to 481) days. Allogeneic blood CD34+ cells can successfully reconstitute hematopoiesis and lymphopoiesis. Reduction of T cells by CD34+ blood cell enrichment and cyclosporine A alone might not be sufficient for prophylaxis of severe acute GvHD.

Topics: Acute Disease; Adult; Antigens, CD34; Bone Marrow; Cell Division; Cyclosporine; Erythropoietin; Female; Filgrastim; Graft vs Host Disease; Granulocyte Colony-Stimulating Factor; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells; Humans; Immunosuppressive Agents; Leukapheresis; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Leukemia, Myeloid; Male; Methotrexate; Middle Aged; Platelet Transfusion; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma; Recombinant Proteins; T-Lymphocytes; Transplantation, Homologous

Core-binding factor leukemia (CBFL) is a subgroup of acute myeloid leukemia (AML) characterized by genetic mutations involving the subunits of the core-binding factor (CBF). The leukemogenesis model for CBFL posits that one, or more, gene mutations inducing increased cell proliferation and/or inhibition of apoptosis cooperate with CBF mutations for leukemia development. One of the most common mutations associated with CBF mutations involves the KIT receptor. A high expression of KIT is a hallmark of a high proportion of CBFL. Previous studies indicate that microRNA (MIR) 222/221 targets the 3' untranslated region of the KIT messenger RNA and our observation that AML1 can bind the MIR-222/221 promoter, we hypothesized that MIR-222/221 represents the link between CBF and KIT. Here, we show that MIR-222/221 expression is upregulated after myeloid differentiation of normal bone marrow AC133(+) stem progenitor cells. CBFL blasts with either t(8;21) or inv(16) CBF rearrangements with high expression levels of KIT (CD117) display a significantly lower level of MIR-222/221 expression than non-CBFL blasts. Consistently, we found that the t(8;21) AML1-MTG8 fusion protein binds the MIR-222/221 promoter and induces transcriptional repression of a MIR-222/221-LUC reporter. Because of the highly conserved sequence homology, we demonstrated concomitant MIR-222/221 down-regulation and KIT up-regulation in the 32D/WT1 mouse cell model carrying the AML1-MTG16 fusion protein. This study provides the first hint that CBFL-associated fusion proteins may lead to up-regulation of the KIT receptor by down-regulating MIR-222/221, thus explaining the concomitant occurrence of CBF genetic rearrangements and overexpression of wild type or mutant KIT in AML.

Topics: AC133 Antigen; Acute Disease; Adolescent; Adult; Aged; Animals; Antigens, CD; Cell Differentiation; Cell Line, Tumor; Cells, Cultured; Core Binding Factor Alpha 2 Subunit; Core Binding Factor alpha Subunits; Down-Regulation; Erythropoietin; Female; Flow Cytometry; Glycoproteins; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Male; MicroRNAs; Middle Aged; Mutation; Oncogene Proteins, Fusion; Peptides; Proto-Oncogene Proteins c-kit; Reverse Transcriptase Polymerase Chain Reaction; RUNX1 Translocation Partner 1 Protein; U937 Cells

Topics: Acute Disease; Anemia, Refractory; Antineoplastic Combined Chemotherapy Protocols; Combined Modality Therapy; Cytarabine; Erythropoietin; Etoposide; Follow-Up Studies; Granulocyte Colony-Stimulating Factor; Humans; Leukemia, Myeloid; Male; Methotrexate; Middle Aged; Mitoxantrone; Peripheral Blood Stem Cell Transplantation; Postoperative Complications; Primary Myelofibrosis; Recombinant Proteins; Transplantation Conditioning; Transplantation, Autologous; Whole-Body Irradiation

Activation of the phosphoinositide 3 kinase (PI3K)/Akt signalling pathway has been linked with resistance to chemotherapeutic drugs, and its downregulation, by means of PI3K inhibitors, lowers resistance to various types of therapy in tumour cell lines. Recently, it has been reported that deguelin, a naturally occurring rotenoid, is a powerful inhibitor of PI3K. We investigated whether or not deguelin could enhance the sensitivity to chemotherapeutic drugs of human U937 leukaemia cells and acute myeloid leukaemia (AML) blasts with an activated PI3K/Akt network. Deguelin (10 nmol/l) induced S phase arrest with interference of progression to G2/M, and at 100 nmol/l significantly increased apoptotic cell death of U937. At 10-100 nmol/l concentrations, deguelin downregulated Akt phosphorylation of leukaemia cells and markedly increased sensitivity of U937 cells to etoposide or cytarabine. A 10 nmol/l concentration of deguelin did not negatively affect the survival rate of human cord blood CD34+ cells, whereas it increased sensitivity of AML blasts to cytarabine. Deguelin was less toxic than wortmannin on erythropoietin- and stem cell factor-induced erythropoiesis from CD34+ progenitor cells. Overall, our results indicate that deguelin might be used in the future for increasing sensitivity to therapeutic treatments of leukaemia cells with an active PI3K/Akt signalling network.

Topics: Acute Disease; Antigens, CD34; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cells, Cultured; Cytarabine; Drug Resistance, Neoplasm; Erythropoietin; Etoposide; HL-60 Cells; Humans; Leukemia; Leukemia, Myeloid; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Rotenone; Signal Transduction; Stem Cell Factor; Stem Cells

A 52-year-old female Jehovah's Witness presented with relapsed secondary acute myeloid leukemia. Because of chemotherapy-induced anemia, she was infused with the bovine hemoglobin (Hb)-based oxygen carrier HBOC-201 (Biopure) as the sole means of transfusion support. HBOC-201 has only been used for management of acute hemorrhage, and its utility in providing longer term transfusion support is unknown. Over a period of 18 days, a total dose of 1230 g of HBOC-201 was delivered. Although the patient succumbed to the disease after 18 days of treatment, this case documents our experience with the highest dose and duration of HBOC-201 ever used. Although possible renal toxicity could not be definitively excluded, the homogeneous extraction of oxygen by the brain in the presence of and perhaps from HBOC-201 was demonstrated.

Topics: Animals; Anti-Bacterial Agents; Antineoplastic Agents; Blood Substitutes; Blood Transfusion; Bone Marrow; Brain; Cattle; Dose-Response Relationship, Drug; Drug Administration Schedule; Drug Therapy, Combination; Erythropoietin; Fatal Outcome; Female; Hemoglobins; Hemosiderin; Humans; Infusions, Intravenous; Jehovah's Witnesses; Leukemia, Myeloid; Middle Aged; Oxygen Consumption; Polymers; Positron-Emission Tomography; Radiography

In our previous study, approximately 60% of aplastic anemia (AA) and refractory anemia (RA) patients treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) and recombinant human erythropoietin (rhEpo) showed a multilineage response. In this study, we analyzed the long-term follow-up of the multilineage responders (multi-R). In the follow-up analysis of 11 multi-R (6 AA and 5 RA), 10 patients (5 AA and 5 RA) were evaluable. The range of time from the start of treatment to the final contact was 50 to 125 months. Analysis of survival times revealed a significant difference between multi-R and non-multi-R among AA patients given this treatment (P = .016). One AA and 1 RA patient among the multi-R developed acute leukemia. Of 7 living multi-R, 3 AA and 2 RA patients did not need transfusion at final contact. Four of them maintained the target hemoglobin concentration of more than 11 g/dL for quality-of-life benefit. The findings suggested that this result is an important advantage of this treatment.

Topics: Acute Disease; Adolescent; Adult; Aged; Anemia, Aplastic; Anemia, Refractory; Blood Transfusion; Erythropoietin; Female; Follow-Up Studies; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Leukemia, Myeloid; Male; Middle Aged; Survival Analysis

We have studied paired peripheral blood progenitor cells (PBPC) and bone marrow (BM) samples from 12 acute myeloid leukaemia (AML) patients following intensive chemotherapy, and assessed direct granulocyte-macrophage colony-forming units (CFU-GM), erythroid burst-forming units (BFU-E), megakaryocyte CFU (CFU-Mk) numbers and the production of CD61+ (platelet glycoprotein IIIa) cells in suspension culture in response to various haemopoietic growth factor combinations. We found that CFU-GM and BFU-E numbers per 105 mononuclear cells were similar in both AML PBPC and BM harvests; CFU-Mk numbers, however, were significantly higher in PBPC than BM. In addition, the higher total white cell count of the PBPC harvests meant that PBPC have much higher numbers of total progenitors per collection. CD61+ cell numbers in suspension cultures of AML PBPC and BM were lower than those of harvested normal marrow. However, response to pegylated recombinant human megakaryocyte growth and development factor (PEGrHuMGDF) both alone and in combination with other growth factors was qualitatively similar to that of normal BM. As with normal BM, response to PEGrHuMGDF alone did not increase further with addition of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), interleukin 6 (IL-6) or erythropoietin (EPO) in the AML PBPC and BM. Further responses over PEGrHuMGDF alone were seen when added with stem cell factor (SCF) or with a combination of SCF + IL-3 + EPO in both AML PBPC and BM cultures; however, the magnitude of the response was greater in the PBPC cultures. Response to PEGrHuMGDF + IL-3 was seen in the PBPC cultures but not in the AML BM. These data suggest that, in AML patients, there are proportionally more megakaryocyte progenitor cells in the mobilized PBPC than in the BM harvests, which would explain the more rapid platelet recovery following PBPC autografts.

Topics: Acute Disease; Adult; Antigens, CD; Blood Platelets; Bone Marrow Cells; Cell Count; Cell Division; Cells, Cultured; Chemokine CXCL12; Chemokines, CXC; Colony-Forming Units Assay; Erythroid Precursor Cells; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Granulocytes; Growth Substances; Hematopoiesis; Humans; Integrin beta3; Interleukin-3; Interleukin-6; Leukemia, Myeloid; Macrophages; Megakaryocytes; Middle Aged; Platelet Membrane Glycoproteins; Polyethylene Glycols; Recombinant Proteins; Stem Cells; Thrombopoietin

We studied the influence of cytokine mixes on the survival of acute myeloid leukemia (AML) bone-marrow (BM) cells in a 14-day culture assay in vitro. Southern-blot analysis using a panel of different probes in combination with densitometry and flow cytometry were used to detect and compare the amount of clonal or CD34-positive BM cells before and after the culturing procedure. A significant reduction of CD34-positive cells after incubation with a cytokine mix [interleukin (IL)-1beta, IL-3, IL-6, stem cell factor (SCF), erythropoietin (EP) with granulocyte macrophage/colony-stimulating factor (GM-CSF, Cytok1) could be achieved in all 16 cases with a CD34-positive blast phenotype studied at diagnosis (P<0.001), in 3 of 10 cases at relapse, and in 8 of 18 cases in complete remission. In healthy donors, an increase of CD34-positive cells was demonstrated in 5 of 5 samples. A reduction of clonal DNA through incubation with Cytok1 was achieved in 5 of 5 (100%) cases studied at diagnosis, in 1 of 4 (25%) cases at relapse, and in 7 of 9 cases (78%) in complete remission. Cytokine cocktails with GM-CSF (Cytok1) were more efficient in reducing (clonal) CD34-positive cells than cocktails without GM-CSF (Cytok2). AML patients at diagnosis and in complete remission had a better survival probability if their CD34-positive or clonal cells could be reduced in vitro by cytokine cultivation (P<0.05). Vitality of BM cells was not influenced by 14-day cytokine treatment; however, the total cell count could be increased by Cytok1 and Cytok2 by 55-174%, but not by the control medium. Our data show that: (1) clonal cell populations can be regularly detected at diagnosis, during complete remission, and at relapse; (2) CD34-positive cells in AML can be demonstrated to be clonal, gene-rearranged cells; (3) incubation of AML BM-cells with Cytokl leads to a reduction of the CD34-positive, clonal cell load in all cases at diagnosis and in 78% of the cases in complete remission of AML, but in only 25% of the cases at relapse; (4) in all healthy BM samples, proportions of 'healthy' CD34-positive cells were increased. Moreover, absolute cell counts were increased by cytokine incubation of cells obtained at diagnosis, relapse, or complete remission of AML and from healthy donors indicating a selective stimulation of healthy, but not of leukemic CD34-positive cells; (5) cytokine cocktails containing GM-CSF are more efficient in reducing leukemic cells than cocktails without GM-CSF; and (6

Topics: Acute Disease; Antigens, CD34; Bone Marrow Cells; Cell Count; Cell Division; Clone Cells; Cytokines; Erythropoietin; Gene Rearrangement; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Stem Cells; Humans; Interleukin-3; Interleukin-6; Leukemia, Myeloid; Prognosis; Recurrence; Remission Induction; Stem Cell Factor

Topics: Acute Disease; Biomarkers; Erythropoietin; Humans; Hypoxia; Leukemia, Myeloid; Neovascularization, Pathologic

P39/Tsugane is a myelomonocytoid cell line derived from a patient with myelodysplastic syndrome (MDS). The cells readily undergo apoptosis in response to various agents, and the cell line has been suggested as a useful model to study apoptosis in MDS. The aims of the present study were to assess differentiation and apoptosis induced with all-trans retinoic acid (ATRA) and etoposide, to characterize the mode of apoptosis in these two model systems, and to assess the influence of granulocyte colony-stimulating factor (G-CSF), which in combination with erythropoietin has been shown to inhibit apoptosis in MDS. ATRA induced differentiation and apoptosis in a concentration- and time-dependent manner. Differentiated cells were partially rescued (by 50%) from apoptosis with G-CSF. Etoposide induced apoptosis in a concentration- and time-dependent manner, but no signs of preceding maturation or G-CSF rescue were detected. ATRA- and etoposide-induced apoptosis were both mediated through the caspase pathway and were partially blocked with the general caspase inhibitor zVAD-fmk. Simultaneous treatment with G-CSF and zVAD-fmk additively blocked ATRA-induced apoptosis. However, the two pathways differed in terms of substrate cleavage during apoptosis. ATRA-induced apoptosis caused actin cleavage, which was not affected by G-CSF, and Bcl-2 downregulation. Etoposide induced a caspase-dependent cleavage of Bcl-2, while actin remained intact. The Fas system did not seem to play a major role in any of these apoptotic pathways. Our results may provide new tools to study the mechanisms of apoptosis in MDS.

Topics: Actins; Acute Disease; Amino Acid Chloromethyl Ketones; Antibodies, Monoclonal; Apoptosis; Blast Crisis; Caspase Inhibitors; Caspases; Cell Differentiation; Cysteine Proteinase Inhibitors; Cytoskeleton; Erythropoietin; Etoposide; fas Receptor; Granulocyte Colony-Stimulating Factor; Humans; Leukemia, Myeloid; Myelodysplastic Syndromes; Neoplasm Proteins; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; Tretinoin; Tumor Cells, Cultured

Our aim was to test the hypothesis that, in leukemic children, serum erythropoietin (EPO) levels vary inversely with hemoglobin.. Twenty-four children (15 males, nine females) with an age range of 1-16 years (mean, 7.7 years) diagnosed with acute leukemia (22 acute lymphocytic, two acute myeloid) were studied over 4 months. Serum EPO and hemoglobin were measured simultaneously at multiple time points in the course of their disease, and a multiple regression analysis was performed to describe the EPO-hemoglobin relationship.. In a model adjusted for individual subject, there was a significant correlation between hemoglobin and logEPO in these leukemic children (r = -0.55, P < .01, n = 100). When measurements at hemoglobins less than 10.0 were analyzed the correlation increased significantly (r = -0.88, P < .01, n = 21). However, approximately 20% of the observations fell into one of two groups: an inappropriately low EPO for hemoglobin or an inappropriately elevated EPO for hemoglobin. The clinical characteristics of the children at each of these determinations were not different in any manner from the determinations which fell within the 95% confidence intervals for predicted mean EPO value: each of the outlying points came from a patient who at other times had an appropriate EPO for hemoglobin.. There existed a significant inverse relationship between hemoglobin and EPO, suggesting that the feedback mechanism for EPO is intact. Reasons for inappropriately high or low EPO, for level of hemoglobin, are not clear and may be reflective of other aspects of bone marrow or EPO metabolism.

Topics: Acute Disease; Biomarkers; Child; Child, Preschool; Erythropoietin; Female; Hemoglobins; Humans; Infant; Leukemia, Myeloid; Male; Precursor Cell Lymphoblastic Leukemia-Lymphoma

RAS mutations arise at high frequency (20-40%) in both acute myeloid leukemia and myelodysplastic syndrome (which is considered to be a manifestation of preleukemic disease). In each case, mutations arise predominantly at the N-RAS locus. These observations suggest a fundamental role for this oncogene in leukemogenesis. However, despite its obvious significance, little is known of how this key oncogene may subvert the process of hematopoiesis in human cells. Using CD34+ progenitor cells, we have modeled the preleukemic state by infecting these cells with amphotropic retrovirus expressing mutant N-RAS together with the selectable marker gene lacZ. Expression of the lacZ gene product, beta-galactosidase, allows direct identification and study of N-RAS-expressing cells by incubating infected cultures with a fluorogenic substrate for beta-galactosidase, which gives rise to a fluorescent signal within the infected cells. By using multiparameter flow cytometry, we have studied the ability of CD34+ cells expressing mutant N-RAS to undergo erythroid differentiation induced by erythropoietin. By this means, we have found that erythroid progenitor cells expressing mutant N-RAS exhibit a proliferative defect resulting in an increased cell doubling time and a decrease in the proportion of cells in S + G2M phase of the cell cycle. This is linked to a slowing in the rate of differentiation as determined by comparative cell-surface marker analysis and ultimate failure of the differentiation program at the late-erythroblast stage of development. The dyserythropoiesis was also linked to an increased tendency of the RAS-expressing cells to undergo programmed cell death during their differentiation program. This erythroid lineage dysplasia recapitulates one of the most common features of myelodysplastic syndrome, and for the first time provides a causative link between mutational activation of N-RAS and the pathogenesis of preleukemia.

Topics: Acute Disease; Antigens, CD34; Apoptosis; Cell Differentiation; Erythroid Precursor Cells; Erythropoiesis; Erythropoietin; Genes, ras; Genes, Reporter; Humans; Leukemia, Myeloid; Mutation; Myelodysplastic Syndromes; Preleukemia

We have established an erythropoietin-dependent human leukemia cell line, AS-E2, from a patient with acute myeloid leukemia. These cells have many characteristics of late erythroid progenitor cells, they are positive for CD36, Glycophorin A, and CD71 but negative for CD41, and positive for benzidine and PAS staining. These cells express GATA-1 and have low affinity erythropoietin (EPO) receptor on their surface. Interestingly, AS-E2 cells are strictly dependent on EPO for their growth and survival; other cytokines including GM-CSF, stem cell factor, or IL-3 cannot support the growth of this cell line. These features are similar to late erythroid lineage cells, like normal BFU-E or CFU-E, and we have demonstrated that EPO stimulation induces the tyrosine phosphorylation of several proteins in AS-E2 cells including the EPO receptor and JAK2 kinase. This new cell line is a useful reagent to study biological and molecular events during the late stages of erythropoiesis, and to understand transforming events in human erythroid cells.

Topics: Acute Disease; Cell Division; DNA Fingerprinting; Erythropoietin; Humans; Karyotyping; Leukemia, Myeloid; Male; Middle Aged; Neoplasm Proteins; Phosphorylation; Receptors, Erythropoietin; Tumor Cells, Cultured; Tumor Stem Cell Assay; Tyrosine

Recombinant human interferon-inducible protein-10 (rIP-10) has been recently identified, purified and shown to suppress the multiplication of normal marrow early hemopoietic progenitors. In the present study we investigated the effect of rIP-10 on different normal and acute myelogenous leukemia (AML) progenitor populations. We first studied hematologically normal bone marrow using the delta culture assay, in which marrow low-density cells were incubated in liquid culture with recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) for 1 week, to allow the differentiation of mature progenitors, and thereafter cultured in methylcellulose in the presence of rGM-CSF and recombinant erythropoietin (rEPO). In this assay rIP-10 significantly inhibited the proliferation of normal marrow hemopoietic progenitors in a dose-dependent fashion. However, when fresh normal marrow cells were cultured in methylcellulose without preincubation in liquid culture, rIP-10 did not affect the growth of colony-forming cells. In contrast, when recombinant c-kit ligand (rKL) was added to rGM-CSF and rEPO, an increment in colony numbers was observed that was eliminated by rIP-10. Similar experiments performed with low-density, non-adherent, T cell-depleted AML marrow cells, obtained from 12 untreated adult AML patients, revealed qualitatively similar results: rIP-10 inhibited the proliferation of AML progenitors in the AML delta assay but did not affect the growth of rGM-CSF-responsive AML colony-forming cells when plated in semisolid media in the presence of rGM-CSF. When rKL was added to rGM-CSF during plating in an effort to recruit additional AML progenitor populations, there was an increment in leukemic blast colony numbers that was eliminated by rIP-10. As observed with normal progenitors, the effect of rIP-10 on these AML progenitors was concentration-dependent, statistically significant and reversible with a rIP-10-neutralizing antiserum. To delineate the mechanism of action of rIP-10 we used the thymidine suicide assay and found that rIP-10 significantly reduced the fraction of leukemic progenitors synthesizing DNA. Our data suggest the rIP-10 inhibits the proliferation of (probably immature) AML progenitor populations by reducing the fraction of cells undergoing DNA synthesis. Additional studies are needed to further elucidate the mechanism of this inhibition and to determine the potential clinical benefits of rIP-10 in future therapies for AML.

Topics: Acute Disease; Aged; Antigens, CD; Antigens, CD34; Antigens, Differentiation, Myelomonocytic; CD13 Antigens; Cell Cycle; Cell Division; Cells, Cultured; Chemokine CXCL10; Chemokines, CXC; Colony-Forming Units Assay; Cytokines; Erythropoietin; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Growth Inhibitors; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Male; Middle Aged; Neoplastic Stem Cells; Recombinant Proteins; Sialic Acid Binding Ig-like Lectin 3; Stem Cell Factor; Tumor Stem Cell Assay

A 34-year-old man with acute myelocytic leukemia (AML: MO) and a 32-year-old woman with AML: M2 developed pure red cell aplasia (PRCA) after receiving a major ABO incompatible bone marrow transplant (BMT). The first patient responded to recombinant human erythropoietin (rhEPO) therapy, while the second did not. The second patient also received methylprednisolone (m-PSL) but developed reticulocytosis and hemolysis after the administration of m-PSL. Plasmapheresis was then performed and the patient promptly recovered from hemolysis and PRCA. We conclude that close attention must be paid when treating PRCA following major ABO-incompatible BMT with rhEPO and m-PSL, as there is always the potential for massive hemolysis.

Topics: ABO Blood-Group System; Acute Disease; Adult; Blood Group Incompatibility; Bone Marrow Transplantation; Erythropoietin; Female; Humans; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Male; Models, Immunological; Recombinant Proteins; Red-Cell Aplasia, Pure; Remission Induction; Transplantation, Homologous

Ineffective hematopoiesis with associated cytopenias and potential evolution to acute myeloid leukemia (AML) characterize patients with myelodysplastic syndrome (MDS). We evaluated levels of apoptosis and of apoptosis-related oncoproteins (c-Myc, which enhances, and Bcl-2, which diminishes apoptosis) expressed within CD34+ and CD34- marrow cell populations of MDS patients (n = 24) to determine their potential roles in the abnormal hematopoiesis of this disorder. Marrow cells were permeabilized and CD34+ and CD34- cells were separately analyzed by FACS to detect: (1) a subdiploid (sub-G1) DNA population, and (2) expression of Bcl-2 and c-Myc oncoproteins. Within the CD34+ subset, a significantly increased percentage of cells demonstrated apoptotic/sub-G1 DNA content in early (ie. refractory anemia) MDS patients compared with normal individuals and AML patients (mean values: 9.1% > 2.1% > 1.2%). Correlated with these findings, the ratio of expression of c-Myc to Bcl-2 oncoproteins among CD34+ cells was significantly increased for MDS patients compared to those from normal and AML individuals (mean values: 1.6 > 1.2 > 0.9). Bcl-2 and c-Myc oncoprotein levels were maturation stage-dependent, with high levels expressed within CD34+ marrow cells, decreasing markedly with myeloid maturation. Treatment of seven MDS patients with the cytokines granulocyte colony-stimulating factor plus erythropoietin was associated with decreased levels of apoptosis within CD34+ marrow cells and may contribute to the enhanced hematopoiesis in vivo that was shown. These findings are consistent with the hypothesis that altered balance between cell-death (eg, c-Myc) and cell-survival (eg, Bcl-2) programs were associated with the increased degrees of apoptosis present in MDS hematopoietic precursors and may contribute to the ineffective hematopoiesis in this disorder, in contrast to decreased apoptosis and enhanced leukemic cell survival in AML.

Topics: Acute Disease; Adult; Aged; Apoptosis; Bone Marrow; Cell Cycle; Cell Transformation, Neoplastic; Disease Progression; DNA, Neoplasm; Erythropoietin; Female; Gene Expression Regulation; Genes, bcl-2; Genes, myc; Granulocyte Colony-Stimulating Factor; Hematopoiesis; Humans; Leukemia, Myeloid; Male; Middle Aged; Myelodysplastic Syndromes; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-myc; Recombinant Proteins

Erythropoietin (Epo) binding to its receptor (EpoR) induces tyrosine phosphorylation in responsive cells and this ability is required for a mitogenic response. One of the substrates of tyrosine phosphorylation is the Epo receptor (EpoR). The carboxyl region of EpoR cytoplasmic domain is required for EpoR phosphorylation and has been shown to negatively affect the response to Epo both in vivo and in cell lines. Hematopoietic cell phosphatase (HCP) has also been hypothesized to negatively regulate erythropoiesis, based on the hypersensitivity to Epo of erythroid lineage cells in moth-eaten mice that genetically lack HCP. In the studies presented here, we show that HCP binds the tyrosine phosphorylated Epo receptor through the amino-terminal src-homology 2 (SH2) domain of HCP. Using a series of phosphotyrosine-containing peptides, potential HCP binding sites in the cytoplasmic domain of the EpoR are identified. The results support the concept that, after Epo stimulation, phosphorylation of EpoR provides a docking site for HCP in the receptor complex. Recruitment of HCP to the complex and its subsequent dephosphorylation of substrates and/or associated kinases may be important to mitigate the ligand-induced mitogenic response.

Topics: Amino Acid Sequence; Animals; Base Sequence; Binding Sites; Cell Line; Disulfides; Erythropoietin; Immunosorbent Techniques; Interleukin-3; Intracellular Signaling Peptides and Proteins; Leukemia, Myeloid; Mice; Molecular Sequence Data; Peptide Fragments; Phosphotyrosine; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Protein Tyrosine Phosphatases; Receptors, Erythropoietin; Recombinant Fusion Proteins; Structure-Activity Relationship; Transfection; Tumor Cells, Cultured; Tyrosine

Using clonogenic assay we investigated the effect of stem cell factor (SCF) on the in vitro growth of clonogenic precursor cells from acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) in the presence or absence of recombinant human erythropoietin (rhEpo) or recombinant human granulocyte colony-stimulating factor (rhG-CSF). SCF as a single factor did not induce significant colony formation, and even in the presence of rhEPO or rhG-CSF it very weakly stimulated erythroid colony formation and was rarely capable of inducing myeloid colony formation by clonogenic leukemic cells. In culture dishes supplemented with SCF, both myeloid and erythroid colony formations were dramatically enhanced in MDS, regarding both colony number and size. Colony-formation abilities by MDS progenitors were improved following costimulation with SCF and rhEpo. These results suggest that SCF may have a therapeutic role in restoring hematopoiesis in patients with MDS.

Topics: Acute Disease; Bone Marrow Cells; Cell Division; Cells, Cultured; Drug Synergism; Erythropoietin; Hematopoiesis; Hematopoietic Cell Growth Factors; Humans; In Vitro Techniques; Leukemia, Myeloid; Myelodysplastic Syndromes; Neoplastic Stem Cells; Stem Cell Factor

A preterm baby, whose mother received chemotherapy for acute leukaemia during pregnancy, required intensive care because of profound anaemia and neutropenia. Haemopoietic progenitor cell studies showed fetal marrow suppression. Those caring for such mothers and babies should know the possible serious effects chemotherapy for malignancies can have on a developing fetus. Long term follow up of the baby is imperative.

Topics: Acute Disease; Adult; Anemia; Antineoplastic Combined Chemotherapy Protocols; Erythropoietin; Etoposide; Female; Granulocyte Colony-Stimulating Factor; Humans; Infant, Newborn; Infant, Premature, Diseases; Leukemia, Myeloid; Neutropenia; Pregnancy; Pregnancy Complications, Neoplastic; Prenatal Exposure Delayed Effects

A novel hematopoietic growth factor, the stem cell factor (SCF), for primitive hematopoietic progenitor cells has recently been purified and its gene has been cloned. In this study, the mitogenic activity of recombinant human SCF on myeloid leukemia cells as well as the expression of its receptor was tested. The proliferation of myeloid leukemia cell lines as well as fresh myeloid leukemic blasts from patients was investigated in a 72 h 3H-thymidine uptake assay in the presence of various concentrations of rhSCF alone or in combination with saturating concentrations of G-CSF, GM-CSF, M-CSF, IL-3, or erythropoietin (EPO). Only five out of 30 lines, but fresh leukemic blasts from 75% of the AML blast samples significantly responded to SCF. To determine the SCF binding sites on leukemic cells, 125I-radiolabelled SCF was used in Scatchard analysis and cross-linking studies. Crosslinking studies demonstrated a 150 kD SCF receptor on the surface of some myeloid leukemic cell lines and all blast preparations. The response to SCF did not correlate to the receptor numbers expressed on the cell surface or to a certain subtype of myeloid leukemia. Using PCR analysis of total RNA from the myeloid leukemia lines we found coexpression of SCF-mRNA and SCF receptor-mRNA in 29% of the myeloid leukemia lines. Soluble as well as membrane bound SCF protein was found to be expressed in myeloid leukemia cells by monoclonal antibodies generated against SCF. This suggests autocrine mechanisms in the growth of a subgroup of leukemic cells by coexpression of SCF and its receptor.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Humans; Interleukin-3; Leukemia, Myeloid; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-kit; Receptors, Colony-Stimulating Factor; Stem Cell Factor

The cytoplasmic domain of the erythropoietin receptor (EpoR) contains a region, proximal to the transmembrane domain, that is essential for function and has homology with other members of the cytokine receptor family. To explore the functional significance of this region and to identify critical residues, we introduced several amino acid substitutions and examined their effects on erythropoietin-induced mitogenesis, tyrosine phosphorylation, and expression of immediate-early (c-fos, c-myc, and egr-1) and early (ornithine decarboxylase and T-cell receptor gamma) genes in interleukin-3-dependent cell lines. Amino acid substitution of W-282, which is strictly conserved at the middle portion of the homology region, completely abolished all the functions of the EpoR. Point mutation at L-306 or E-307, both of which are in a conserved LEVL motif, drastically impaired the function of the receptor in all assays. Other point mutations, introduced into less conserved amino acid residues, did not significantly impair the function of the receptor. These results demonstrate that conserved amino acid residues in this domain of the EpoR are required for mitogenesis, stimulation of tyrosine phosphorylation, and induction of immediate-early and early genes.

Topics: Amino Acid Sequence; Animals; Cell Division; Cell Line, Transformed; Cytokines; Enzyme Induction; Erythropoietin; Interleukin-3; Leukemia, Myeloid; Molecular Sequence Data; Mutagenesis, Site-Directed; Phosphorylation; Point Mutation; Protein-Tyrosine Kinases; Receptors, Cell Surface; Receptors, Erythropoietin; Sequence Homology, Amino Acid; Structure-Activity Relationship; Transcription, Genetic

Phenylalanine methylester (PME), a lysosomotropic compound can be used to deplete monocytes and myeloid cells from peripheral blood and bone marrow (BM). The potential of PME for purging leukemic cells from BM was investigated using U937 and HL-60 cell lines as models. Optimal purging conditions for U937 cells were determined using an MTT assay (3-4, 5-dimethylthiazol-2, 5-diphenyl tetrazolium biomide; Sigma). Elimination of U937 cells was time-, temperature-, and dose-dependent. PME activity was optimal at 37 degrees C for 45 minutes. Depletion of U937 was > 2.8 logs for 50 mmol/L PME. Compared with another purging agent, 100 micrograms/mL 4-hydroperoxycyclophosphamide had activity comparable to 40 mmol/L PME. HL-60 cells were even more sensitive to PME than U937 cells. To support observations made with the MTT assay, clonogenic assays were performed. PME, 50 mmol/L at 37 degrees C resulted in total depletion (> 5 logs) of U937 colonies. Progressive depletion of normal progenitor cells occurred when BM was incubated with PME at concentrations from 5 to 100 mmol/L. At 37 degrees C, 50 mmol/L PME reduced colony-forming units-granulocyte-macrophage and burst-forming units-erythroid (BFU-E) recovery by 98%. Recombinant human mast cell factor augmented BFU-E after PME treatment but had no effect on HL-60 or U937. These studies suggest that PME deserves further study as an agent for ex vivo marrow purging.

Topics: Bone Marrow; Bone Marrow Purging; Colony-Forming Units Assay; Cyclophosphamide; Erythropoietin; Hematopoietic Cell Growth Factors; Humans; Kinetics; Leukemia, Myeloid; Leukemia, Promyelocytic, Acute; Phenylalanine; Recombinant Proteins; Stem Cell Factor; Time Factors; Tumor Cells, Cultured

A proportion of fetal liver hemopoietic blast cells express Fc gamma RII, and addition of the anti-Fc gamma RII monoclonal antibody CIKM5 induces a rise in calcium in these cells in suspension. Although these cells are thus capable of mobilizing intracellular calcium in response to surface receptor mediated events, neither granulocyte-macrophage colony-stimulating factor (GM-CSF) nor erythropoietin produced detectable changes in intracellular calcium ion concentration in these cells.

Topics: Antibodies, Monoclonal; Antigens, Differentiation; Calcium; Cell Line; Erythropoietin; Fetus; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoiesis; Hematopoietic Stem Cells; Humans; Immunoenzyme Techniques; Immunophenotyping; Leukemia, Erythroblastic, Acute; Leukemia, Myeloid; Leukemia, Promyelocytic, Acute; Receptors, Fc; Receptors, IgG; Tumor Cells, Cultured

Some mouse myeloid leukemias induced by X-irradiation and serially transplanted into syngenic mice do not proliferate in vitro even in the presence of hematopoietic factors. To examine whether such leukemic cells can proliferate in response to stromal cells, we cocultured them with MC3T3-G2/PA6 (PA6) preadipocytes, cells that can support the growth of hematopoietic stem cells. All leukemias developed into in vitro cell lines, showing a dependence on contact with the PA6 cells. Two cell lines responded to none of the known hematopoietic factors including interleukin-3 (IL-3), IL-4, IL-5, IL-6, GM-CSF, G-CSF, M-CSF, and Epo. These results demonstrate that the mechanism of the action of PA6 cells is different from that of any of the known hematopoietic factors, and that, because these two leukemic cell lines retained the ability to grow in vivo, responsiveness to the known hematopoietic factors is not essential for the leukemic cell growth in vivo. Furthermore, all leukemic cell lines could respond also to the preadipocytes fixed with formalin, paraformaldehyde, or glutaraldehyde, suggesting that some molecule(s) associated with the surface of PA6 cells or with extracellular matrix secreted by the preadipocytes is responsible for the leukemic cell growth.

Topics: Adipose Tissue; Animals; Cell Communication; Cell Division; Cell Line; Erythropoietin; Female; Formaldehyde; Glutaral; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Interleukins; Leukemia, Experimental; Leukemia, Myeloid; Leukemia, Radiation-Induced; Macrophage Colony-Stimulating Factor; Male; Mice; Mice, Inbred C3H; Polymers; Recombinant Proteins

Bone marrow and peripheral blood cells from a patient with chronic myelogenous leukemia in erythroblastic transformation were studied by flow cytometry and for hemopoietic colony growth. Results demonstrated that this disorder had greatly expanded bone marrow erythroid colony (CFU-E) and myeloid colony (CFU-GM) progenitor compartments that were totally dependent upon erythropoietin and colony-stimulating factor. DNA, RNA and cell cycle analysis revealed that the bone marrow cells were diploid, had a high percentage of S phase cells (17%), and a unique bimodal RNA index of 5 and 13.8. Results are discussed and contrasted with other myeloproliferative disorders.

Topics: Adult; Bone Marrow Cells; Cell Division; Cells, Cultured; Colony-Forming Units Assay; Colony-Stimulating Factors; DNA; Erythroblasts; Erythrocytes; Erythropoietin; Flow Cytometry; Granulocytes; Hematopoiesis; Humans; Leukemia, Myeloid; Macrophages; Male; RNA

A patient with Ph1-positive chronic myelogenous leukemia (chronic phase) had a cyclic oscillation in white blood cells, platelets and percent saturation of transferrin. The cycle comprised about 70 days. The number of circulating granulocyte-macrophage colony-forming units (CFU-GM) oscillated with the same phase, while that of bone marrow CFU-GM and erythroid colony-forming units (CFU-E) oscillated in a reverse phase. At the nadir, we observed an abnormal increase in bone marrow endogenous CFU-E (e-CFU-E). An erythropoietin (Epo) dose-response curve of CFU-E showed a high Epo-sensitivity. Anti-Epo rabbit serum did not inhibit the e-CFU-E colony formation. This indicates that Epo-independent erythropoiesis occurs periodically at the nadir. It is suggested that the interactions between the abnormal stem cell and the hematopoietic regulating system cause cyclic oscillation.

Topics: Adult; Bone Marrow; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Leukocyte Count; Male; Periodicity; Platelet Count

Topics: Adult; Erythropoietin; Fetal Hemoglobin; Hematopoietic Stem Cells; Humans; In Vitro Techniques; Leukemia, Myeloid

Clone cells of K562 that are able to synthesize hemoglobin spontaneously on a relatively high level were obtained by cell-cloning technique. The clone cell proliferated 25 times by day 6 in culture, and the growth rate was not affected by changing the dose of fetal calf serum (FCS) from 5% to 30%. On the other hand, the erythroid differentiation could be linearly enhanced by increasing dosage of FCS, reaching a maximum after four days in culture. The wild-type K562 cells were also slightly stimulated to synthesize hemoglobin by adding FCS (30% final concentration). The enhancing effect of 30% FCS on the erythroid differentiation in the clone cells was greater than that of 12.5 microM hemin, while in the wild-type cells the relationship was reversed. There were no effects of erythropoietin (Epo) on the hemoglobin synthesis in either the clone cells or the wild-type cells. When various kinds of sera were added to the standard culture of the clone cells, only FCS had the enhancing effect. These results suggest that spontaneous erythroid differentiation is not induced by hemin or Epo in FCS but by FCS-specific substance(s).

Topics: Adult; Animals; Blood; Cattle; Cell Differentiation; Cell Line; Clone Cells; Erythrocytes; Erythropoietin; Fetal Blood; Hemin; Horses; Humans; Infant, Newborn; Leukemia, Myeloid; Time Factors

Peripheral blood cells from 2 patients with chronic granulocytic leukemia were separated by density centrifugation. Mononuclear cells of low density (d less than 1.062g cm-3) with blast-cell morphology were cryopreserved before culture in vitro. Upon culture in conventional colony assays, up to 20% of the cells formed hemopoietic colonies. Although the spectrum of colony types resembled that of normal bone-marrow cells, there were large differences between the patients with respect to the number, type and size of colonies that were observed. Colony formation required the addition of hemopoietic growth factors, such as colony-stimulating activity and erythropoietin to the culture medium. The cells were used to assay hemopoietic regulatory molecules. Both erythropoietin and colony-stimulating activity induced a strong proliferative response as measured by thymidine incorporation. Maximal stimulation was observed when erythropoietin and the supernatant of mixed lymphocyte cultures were added simultaneously. The difference between the cells from the 2 patients in clonal assays was reflected by the different response to individual hemopoietic regulators. The time course of maximal stimulation followed distinct patterns dependent on the source of stimulator. The stimulation was linearly dependent on the input cell number. Taken together, cryopreserved blast cells from patients with chronic granulocytic leukemia appear to be very useful for the characterization of factors regulating hemopoiesis, as well as for studies of hemopoiesis in general.

Topics: Cell Division; Cell Separation; Colony-Forming Units Assay; Colony-Stimulating Factors; Erythropoietin; Growth Substances; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Phytohemagglutinins

Serum erythropoietin (ESF) levels and the numbers of marrow and blood erythroid progenitors (CFU-Es) of patients with chronic myeloproliferative disorders (CMPD) were studied simultaneously. The numbers of marrow and blood CFU-Es per 1 x 10(5) cells were normal or greatly elevated. There was an inverse correlation between the hemoglobin concentration and the serum ESF level in patients with chronic myelogenous leukemia when the hemoglobin concentration ranged from 9.0 to 13.0 g/100 ml. The serum ESF level was closely related to the hemoglobin concentration in CMPD and it was suggested that the negative feedback mechanism might operate in anemic patients with CMPD.

Topics: Adult; Animals; Bone Marrow Cells; Cell Count; Chronic Disease; Erythropoietin; Female; Hematopoietic Stem Cells; Hemoglobins; Humans; Leukemia, Myeloid; Male; Mice; Middle Aged; Myeloproliferative Disorders; Polycythemia Vera; Primary Myelofibrosis; Thrombocytosis

Topics: Child, Preschool; Erythroblasts; Erythropoiesis; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; In Vitro Techniques; Leukemia, Myeloid

Early erythroid progenitors (BFUE) form colonies of mature progeny in culture. The development of hemoglobinized red cells within multilineage colonies (CFUGEMM) and erythroid bursts is dependent upon exogenously added erythropoietin and molecules released by hemopoietic subpopulations. Mixed colonies and erythroid bursts were grown from 3 patients with Ph' chronic myelogenous leukemia (CML). It was found that some mixed hemopoietic colonies and erythroid bursts did not require exogenously added erythropoietin. An increase of the plating efficiency of BFUE could be observed when erythropoietin was added. Erythroid bursts grown without added Ep from samples of the patients with chronic myelogenous leukemia have a higher probability to contain HbF than clones grown in the presence of Ep. The data support the view of a phenotypical heterogeneity among clonal descendents of a common ancestor as previously postulated for CML.

Topics: Adult; Bone Marrow Cells; Erythrocytes; Erythropoietin; Fetal Hemoglobin; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid

Human hemopoietic pluripotent progenitors form multilineage colonies when cultured in methylcellulose with medium conditioned by leukocytes in the presence of phytohemagglutinin (PHA-LCM) and erythropoietin (EPO). We have examined their frequency, culture requirements and proliferative activity in 20 peripheral blood and 29 bone marrow specimens from patients with CML in chronic phase. Multilineage colonies developed under regular culture conditions in approximately 50% of all samples. The frequency ranged from 1-36 per 2 X 10(5) mononuclear cells of density less than 1.077 gm/ml. The requirements for PHA-LCM and EPO varied for patients with CML when compared to normal individuals; i.e., cells from some patients gave rise to mixed colonies with substantial erythroid components in the absence of PHA-LCM or without addition of the usually required EPO concentrations. The proliferative activity of CFU-GEMM was assessed using a short-term exposure to tritiated thymidine (3HTdR) prior to plating. The plating efficiency in all bone marrow and peripheral blood samples was reduced to 40-70% of the unexposed controls. In contrast, the plating efficiency after exposure to 3HTdR in normal individuals usually ranged from 70-90% of controls for bone marrow samples and from 85-100% of controls for peripheral blood samples. Thus, an increased proliferative rate of pluripotent hemopoietic progenitors is a consistent feature of CML patients. In addition, at least in some patients, different requirements for erythropoietin or PHA-LCM were observed when compared to normal culture conditions.

Topics: Cell Division; Cells, Cultured; Colony-Forming Units Assay; Culture Media; Erythropoietin; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Leukocytes; Phytohemagglutinins

Topics: Bone Marrow Cells; Cells, Cultured; Chromosomes, Human, 21-22 and Y; Colony-Forming Units Assay; Demecolcine; Erythrocytes; Erythropoietin; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Monocytes

The requirement of erythropoiesis for erythropoietin were studied in a patient with Ph chronic myelocytic leukemia who had undergone an erythroblastic transformation. Transfusions resulted in a suppression of erythropoiesis. Plasma clot culture studies indicated that both the CFU-E and BFU-E in the peripheral blood of this patient were dependent upon erythropoietin for their differentiation and proliferation. Neither of these committed erythroid stem cells was cloned in the absence of erythropoietin. These studies suggest that the proliferation and differentiation of erythroid stem cells during the erythroblastic crisis of this disorder remain dependent upon physiologic regulators.

Topics: Adult; Blood Transfusion; Chromosomes, Human, 21-22 and Y; Erythrocyte Transfusion; Erythropoiesis; Erythropoietin; Female; Humans; Leukemia, Myeloid

Myeloproliferative virus, derived from Moloney sarcoma virus, causes erythroleukemia and myeloid leukemia in adult mice. This virus is also capable of fibroblast transformation in vitro. The virus consists of two separable biological entities which have been cloned. The helper virus component caused no visible changes in adult mice, whereas the defective virus induced both spleen focus formation and a large increase in erythroid precursor cells but retained the sarcoma virus property of transforming fibroblasts in vitro. Thus, myeloproliferative virus is the first murine sarcoma virus which induces erythroleukemia in adult animals.

Topics: Animals; Bone Marrow Cells; Cell Transformation, Viral; Cells, Cultured; Defective Viruses; Erythropoietin; Fibroblasts; Helper Viruses; Leukemia, Erythroblastic, Acute; Leukemia, Experimental; Leukemia, Myeloid; Mice; Mice, Inbred Strains; Rats; Sarcoma Viruses, Murine; Spleen; Virus Cultivation

A patient with Ph1-positive chronic myelogenous leukemia (CML) who entered an erythroblastic transformation prior to the development of a typical myeloblastic crisis is described. In vitro methylcellulose cultures obtained at the time of erythroblastic transformation revealed that the erythroid progenitors were responsive to erythropoietin. Thus, similar to the findings in polycythemia vera, the erythroid progenitors in this case of erythroblastic transformation of CML retained responsiveness to erythropoietin in vitro.

Topics: Bone Marrow; Bone Marrow Cells; Cell Transformation, Neoplastic; Cells, Cultured; Culture Media; Erythroblasts; Erythrocytes; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Methylcellulose; Middle Aged

Peripheral blood and bone marrow specimens from patients with polycythemia vera (PV) and chronic myelogenous leukemia (CML) were assayed for erythroid and granulopoietic progenitor cells. All compartments were increased in CML patients in relapse although the ratio of BFU-E to CFU-C numbers remained constant in all CML patients where values ranged over several orders of magnitude. By comparison with normal ratios there was only a slight shift towards increased CFU-C numbers. No quantitative changes in any progenitor compartment was found in PV except for a marginal increase in marrow CFU-E. Erythropoietin (epo)-independent colony formation has been documented in all 61 cases of PV studied to date, and the proportion of progenitors classified as abnormal on this basis increases on average 3- to 5-fold as they differentiate in vivo from primitive BFU-E to CFU-E. Preliminary replating studies suggest that when this occurs in vitro individual BFU-E produce both normal and abnormal phenotypes. Epo-independent erythropoiesis has also been commonly observed in assays of CML cells, although its expression is more variable and in the absence of epo progenitors in CML usually make fewer erythroblasts containing even less hemoglobin than do their counterparts in PV. Expression of a common regulatory defect in erythroid cells in PV and CML suggests a possible relationship to the initial transformation event(s).

Topics: Antibody Formation; Colony-Forming Units Assay; Erythropoiesis; Erythropoietin; Hematopoietic Stem Cells; Humans; Immunity, Cellular; Leukemia, Myeloid; Polycythemia Vera

Serum of patients suffering from a chronic myeloproliferative disorder (polycythaemia, era, osteomyelofibrosis, chronic myeloid leukaemia) and serum of lethally irradiated rats injected before application of a single doses of erythropoietin did not enhance the effect of erythropoietin -- measured with the iron incorporation rate of polycythemic mice. The rationale for these experiments is to try to find a "myeloproliferative factor", which augments the number of stem cells as described in sera of patients with polycythaemia vera, osteomyelofibrosis, and lethally irradiated mice.

Topics: Adult; Aged; Drug Synergism; Erythropoietin; Humans; Leukemia, Myeloid; Middle Aged; Myeloproliferative Disorders; Polycythemia Vera; Primary Myelofibrosis

The ability of cells derived from the K562 cell line to generate erythropoietic colonies was studied. The K562 cell line was derived from a patient with chronic myelogenous leukemia 8 yr ago by Lozzio and Lozzio. Rare benzidine-positive colonies formed when these cells were cloned in plasma clots (3 +/- 1/10(4) cells), and their number was not substantially increased by the addition of erythropoietin (9.5 +/- 1/10(4) cells). Sodium butyrate was capable of markedly enhancing the number of benzidine-positive colonies (19.5 +/- 1/10(4) cells) formed, while the combination of sodium butyrate plus erythropoietin exerted a synergistic effect on erythropoietic colony formation (57 +/- 4/10(4) cells). The K562 cell line is a long-term culture system that contains human erythropoietic stem cells. This cell line should be useful in future studies on the cellular and molecular events associated with human erythroid cell differentiation.

Topics: Benzidines; Butyrates; Cell Line; Cells, Cultured; Dimethyl Sulfoxide; Erythropoiesis; Erythropoietin; Humans; Leukemia, Myeloid; Time Factors

Peripheral blood and bone marrow specimens from 6 patients with Ph1-positive CML were evaluated for their content of erythropoietic and granulopoietic colony-forming progenitor cells. Specimens obtained from untreated patients showed marked increases in all compartments the most dramatic of which were for CFU-E and circulating CFU-C. This increased cell flow down the early stages of the red cell pathway in CML suggests that heightened proliferation and differentiation of primitive hemopoietic cells may be a more general phenomenon than previously suspected in this disease. In 5 of 6 patients, abnormal erythroid progenitors capable of proliferation and differentiation into hemoglobinized erythroblasts in cultures containing less than 0.002 units of erythropoietin/ml were regularly detected. In the 6th patient abnormal growth was not seen in cultures of the initial marrow obtained but was detected in cultures set up with peripheral blood taken 7 months later. The unexplained amplification of the erythropoietic compartment and the ability of some of these cells to mature in vitro in the virtual absence of erythropoietin is at variance with the anemia characteristic of untreated patients. This suggests the possibility of a major differentiation block at the level of CFU-E. Further studies of the properties of erythroid progenitors in these patients should help to provide new insights into the pathogenesis of CML and may provide useful markers for monitoring engrafted cell function after autotransplantation of patients in blast crisis.

Topics: Adult; Aged; Colony-Forming Units Assay; Erythropoiesis; Erythropoietin; Female; Granulocytes; Hematopoiesis; Hematopoietic Stem Cells; Humans; Leukemia, Myeloid; Male; Middle Aged

Fetal mouse liver cultures capable of producing both erythropoietin (Ep) and granulocyte-macrophage colony stimulating activity (GM-CSA) were used to study the specificity of lactoferrin as an inhibitor of the production of GM-CSA. Both a granulocyte-derived colony-inhibiting activity (CIA) and lactoferrin inhibited GM-CSA production while having no effect on Ep production. These results demonstrate the specificity of lactoferrin for GM-CSA production.

Topics: Animals; Colony-Stimulating Factors; Erythropoietin; Female; Fetus; Granulocytes; Lactoferrin; Lactoglobulins; Leukemia, Myeloid; Liver; Macrophages; Mice; Neutrophils; Transferrin

Erythropoietin (Ep) levels were measured in Shay chloroleukemic rats at various stages of anemia. Serum Ep was shown to increase logarithmically as the anemia became more severe. This increase in Ep levels was similar to that observed in normal rats subjected to acute blood loss. Significant levels of Ep were also demonstrated in ascitic fluid extracted from the peritoneal cavity of leukemic rats. These results indicate that the anemia of this disease is not due to a diminished production of Ep.

Topics: Anemia; Animals; Ascitic Fluid; Erythropoietin; Leukemia, Experimental; Leukemia, Myeloid; Male; Rats

A child aged 3 1/2 years with typical juvenile chronic granulocytic leukaemia whose course terminated in an erythroblastic phase is described. Both in-vivo transfusion studies and in-vitro bone marrow culture studies indicate that red cell production remains erythropoietin dependent in this situation. The relationship of erythropoietin to the reversion to fetal haematopoiesis which characterizes juvenile chronic granulocytic leukaemia is discussed.

Topics: Child, Preschool; Erythroblasts; Erythropoiesis; Erythropoietin; Female; Humans; In Vitro Techniques; Leukemia, Myeloid

A spontaneous oscillation of the white blood cell count was observed in a 58 year old man with chronic myelogenous leukemia (CML). Similar cyclic variations were noted in the platelet and reticulocyte counts with no apparent alterations in marrow cellularity to account for such changes. Since direct correlation was noted between white blood cells, platelets, and reticulocyte counts versus spleen size, it suggests that splenic hemopoiesis may be responsible for these cyclic changes. A possible inverse relationship between colony-stimulating factor (CSF) activity and the white blood cell count was noted, suggesting that CSF may be the humoral agent controlling granulocyte production. A direct correlation between the white blood cell count and serum unsaturated vitamin B12 binding capacity (UBBC) and lysozyme was also noted and further supports the concept that the latter two are measures of the granulocyte pool and metabolism. An inverse relationship between CSF activity and the UBBC suggests that these may be two different entities. Finally a modified form of standard chemotherapy may be effective in inducing remission in cases of CML with marked cyclic leukocytosis-leukopenia.

Topics: Alkaline Phosphatase; Blood Cell Count; Blood Platelets; Bone Marrow Examination; Colony-Stimulating Factors; Erythropoietin; Hemoglobins; Humans; Karyotyping; Leukemia, Myeloid; Leukocyte Count; Leukocytosis; Leukopenia; Male; Middle Aged; Muramidase; Periodicity; Reticulocytes; Spleen; Vitamin B 12

Topics: Animals; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Erythropoietin; Leukemia, Experimental; Leukemia, Myeloid; Lymphocytes; Male; Rats; RNA; Spleen; Tritium; Uridine

Topics: Adult; Aged; Animals; Blood Cell Count; Blood Platelets; Blood Transfusion; Erythropoietin; Female; Hematocrit; Hematopoietic Stem Cells; Humans; Injections, Subcutaneous; Iron; Iron Radioisotopes; Leukemia, Myeloid; Leukocyte Count; Leukocytosis; Male; Mice; Middle Aged; Polycythemia Vera; Primary Myelofibrosis

Two patients with chronic myelocytic leukemia who developed an erythroblastic rather than a myeloblastic phase were studied with respect to whether or not the megaloblastic erythropoiesis was subject to normal control mechanisms. After transfusion, no significant reduction was observed in the percentage of nucleated erythroid precursors or of proerythroblasts in marrow or in blood reticulocytes. In one of the two patients, ferrokinetics and urinary erythropoietin levels were studied and were also compatible with the conclusions that erythropoiesis was autonomous in this rare syndrome. Three patients with clinical pictures compatible with Di Guglielmo's syndrome were studied as controls. As has been reported previously, erythropoiesis in this syndrome appeared to be responsive to normal control mechanisms. These data suggest that these two clinically similar syndromes, erythroblastic crisis of chronic myelocytic leukemia and Di Guglielmo's syndrome may represent qualitatively different defects in hematopoietic stem cells.

Topics: Adult; Aged; Blood Transfusion; Erythrocytes; Erythropoiesis; Erythropoietin; Female; Humans; Iron; Leukemia, Myeloid; Male; Middle Aged

Topics: Adult; Aged; Animals; Erythropoiesis; Erythropoietin; Female; Humans; Leukemia, Myeloid; Male; Mice; Middle Aged

Topics: Acute Disease; Adolescent; Child; Child, Preschool; Erythropoiesis; Erythropoietin; Hodgkin Disease; Humans; Infant; Leukemia; Leukemia, Myeloid

Topics: Binding Sites, Antibody; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Culture Techniques; Erythropoietin; Hematopoiesis; Hematopoietic Stem Cells; Humans; Leukemia, Lymphoid; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Macrophages; Methods; Microscopy, Electron; Mitosis; Multiple Myeloma; Neoplasms; Phagocytosis; Thymidine; Tritium; Waldenstrom Macroglobulinemia

Topics: Anemia, Aplastic; Bone Marrow; Bone Marrow Cells; Erythropoiesis; Erythropoietin; Humans; Leukemia, Myeloid

Topics: Adult; Aged; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Erythropoiesis; Erythropoietin; Female; Hematocrit; Heme; Hemoglobinometry; Humans; Iron; Iron Isotopes; Leukemia, Myeloid; Male; Middle Aged; Polycythemia Vera; Reticulocytes

Topics: Adolescent; Adult; Aged; Blood Platelets; Bone Marrow Diseases; Diagnosis, Differential; Erythrocyte Count; Erythropoietin; Female; Hematocrit; Hemoglobinometry; Humans; Leukemia, Myeloid; Leukocyte Count; Male; Middle Aged; Myeloproliferative Disorders; Polycythemia Vera; Primary Myelofibrosis; Retrospective Studies

Topics: Animals; Erythropoietin; Hemoglobinometry; Humans; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid; Mice

Topics: Bone Marrow; Epoetin Alfa; Erythropoietin; In Vitro Techniques; Leukemia; Leukemia, Myeloid; Monocytes; Pharmacology; Research; Synovial Membrane; Tissue Culture Techniques