Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Erythroid Precursor Cells”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 793 records · Page 44Linked to original sources

Granulocyte colony stimulating factor: from laboratory bench to clinical use.

A great deal has been learned about haematopoiesis since the discovery, purification and cloning of haemopoietic growth factors. The function of these growth factors is to promote survival and enhance mitosis (at higher concentrations), and influence differentiation and development of stem cells and progenitor cells. The type of mature cells produced reflects the combinations of growth factors to which the stem cells are exposed. Growth factors may also control the functional activity of mature cells. Treatment with granulocyte colony stimulating factor (G-CSF) produces neutrophils that are at least equivalent to those produced during normal haematopoiesis and does not appear to deplete the haematopoietic stem or progenitor cell populations.

Animals↗

The changing face of stem cell transplantation by the use of recombinant human granulocyte colony stimulating factor.

The use of peripheral blood progenitor cells (PBPC) for autografting following high-dose chemotherapy in a variety of malignancies has increased markedly with the advent of efficacious and safe myeloid growth factors, which have dramatically improved collection yields. There is still controversy as to whether PBPC should be harvested after a combination of chemotherapy and growth factors, or after growth factors alone. The use of powerful combinations of newer growth factors, such as interleukin-3 with granulocyte-macrophage colony stimulating factor (GM-CSF), or granulocyte colony stimulating factor (G-CSF) with stem cell factor, may obviate the need for chemotherapy. The problem of providing sufficient numbers of PBPC for successful transplantation and sustained engraftment has led to the development of a variety of in vitro expansion systems for stem cells, using cocktails of growth factors. Single blood collections may contain sufficient CD34+ cells to be expanded on a large scale for clinical transplantation, eliminating costly and labour intensive apheresis procedures. PBPC transplants may carry less risk for contamination with malignant cells; however, sensitive detection techniques have revealed that tumour cell contamination of PBPC harvests may be much more prevalent than was previously suspected and both positive and negative selection of CD34+ cells for clinical transplantation is being investigated. PBPC transplantation generally results in more rapid engraftment, with faster recovery of neutrophils and platelets, compared with autologous bone marrow transplants. In most studies, PBPC transplants also resulted in fewer septic episodes, fewer intensive care admissions, fewer transfusions of red blood cells and platelets, reduced antibacterial and parenteral nutrition use, and reduced hospital costs. Dose optimisation and intensification of chemotherapy, relying on repeated administration of growth factor-mobilised PBPC, offers interesting prospects in the treatment of a variety of tumours. However, well-controlled, randomised prospective trials will be needed to prove the real value of PBPC transplants with regard to survival and cure. In the clinical setting, PBPC transplantations are likely to replace autologous marrow transplants in the near future. Manipulation of PBPC in vitro, i.e. expansion or gene transfer, may prove to be the most exciting perspective in the treatment of both malignant and non-malignant haematological conditions.

Cell Division↗

IL-12 directly enhances in vitro murine erythropoiesis in combination with IL-4 and stem cell factor.

It has been demonstrated recently that in vivo administration of murine IL-12 to mice enhances the activity of cytotoxic NK cells and lymphocyte-activated killer cells, and that it has antitumor and antimetastatic activity. However, one side effect observed in response to systemic IL-12 treatment is anemia. In the present study, we examined for the first time the ability of IL-12 to affect directly the growth of murine erythroid progenitor cells in vitro. Whereas IL-12 alone or in combination with Erythropoietin (Epo) showed no stimulatory effect on erythroid progenitors, IL-12 potently enhanced the number of erythroid burst-forming unit (BFU-E) colonies formed in response to Epo+IL-4 by 63% and Epo+stem cell factor by 80%. The stimulatory effect of IL-12 occurred in a concentration-dependent fashion, with maximum enhancing effect observed at 50 ng/ml. Furthermore, single cell experiments suggested that the stimulatory effect of IL-12 on erythroid colony formation was directly mediated. Thus, IL-12 can directly enhance murine erythropoiesis in vitro, suggesting that IL-12-induced anemia is mediated through an indirect mechanism.

Animals↗

Differentiation-associated changes in CD44 isoform expression during normal hematopoiesis and their alteration in chronic myeloid leukemia.

CD44 is a widely expressed, multifunctional, cell-surface glycoprotein that has been implicated in the regulation of normal hematopoiesis. In addition, expression of particular isoforms of CD44 has been associated with malignant transformation and/or the acquisition of metastatic potential. In this study, we used two recently developed monoclonal anti-CD44 antibodies, one reactive with an epitope shared by many CD44 isoforms and the other with an epitope unique to CD44 isoforms containing amino acids encoded by the alternatively spliced exon v10, to compare the expression of CD44 on primitive hematopoietic cells from the marrow of normal individuals and their neoplastic counterparts present in the peripheral blood of patients with chronic myeloid leukemia (CML). Multiparameter fluorescence-activated cell sorter (FACS) analysis and cell sorting studies showed that CD44 is normally expressed at high to very high levels on both long-term culture-initiating cells (LTC-IC) and granulopoietic colony-forming cells (granulocyte-macrophage colony-forming units [CFU-GM]). In contrast, primitive erythropoietic progenitors (burst-forming units-erythroid [BFU-E]) in normal marrow were more homogeneous in their expression of CD44, and very few (less than 5%) showed the very high levels of CD44 seen on 20% to 25% of LTC-IC and CFU-GM. Antibody staining showed the expression of exon v10-containing CD44 isoforms to be restricted to a small subpopulation (4% to 8%) of morphologically recognizable mature (CD34-) myeloid cells within the light-density fraction of normal marrow cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed the presence of two exon v10-containing mRNA species. In CML, a significantly greater proportion of the circulating neoplastic CFU-GM expressed very high levels of CD44, and these CFU-GM were accompanied by an increased number of light density v10+ cells, including some that coexpressed CD34. Nonmalignant hematopoietic progenitors mobilized by prior chemotherapy and growth factor treatment of patients with Hodgkin's disease or acute myeloid leukemia in remission showed no changes in CD44 expression relative to normal marrow progenitors. These results provide evidence of early differentiation-associated changes in CD44 expression during normal hematopoiesis in vivo that may be deregulated in the neoplastic clone of patients with CML.

Base Sequence↗

[Effect of macrophage inflammatory protein-1 alpha (MIP-1 alpha) on human erythropoiesis in vitro--clinical implications].

The influence of macrophage inflammatory protein-1 alpha (MIP-1 alpha) on cloning efficiency of human erythroid progenitors was evaluated. In our hands, in performed experiments in serum supplemented as in serum free cloning culture systems we did not observe any effect of MIP-1 alpha on erythroid colony formation by human CD34+ bone marrow cells. Therefore, our data did not confirm the possibility of the direct role of MIP-1 alpha in the pathogenesis of the anaemia of chronic disease.

Bone Marrow↗

[T lymphocytes from patients with Hodgkin's disease suppress erythroid progenitor growth from autologous marrow].

The effect of peripheral blood T lymphocytes from 42 patients with advanced Hodgkin's disease (grade III and IV) on the autologous marrow erythroid colony formation was studied in diffusion chamber culture. It was found, that unfractionated T lymphocytes suppress the BFU-E--(burst forming unit erythroid) and CFU-E--(colony forming unit erythroid)--derived colony formation by releasing an inhibitory activity. The suppression of colony formation was noted already at 0.25 x 10(5) cell concentration. In experiments with 0.5 x 10(5) and 1.0 x 10(5) T cells the inhibitory effect was increased. Subsequently it was shown, that this inhibition was generated by radioresistant, CD8+ and HLA-DR- subset of T cells. In control experiments, T lymphocytes from healthy subjects had no influence on the erythroid colony formation.

Adolescent↗

Influence in vitro of IL-3/Epo fusion proteins compared with the combination of IL-3 plus Epo in enhancing the proliferation of single isolated erythroid and multipotential progenitor cells from human umbilical cord blood and adult bone marrow.

Human interleukin-3/erythropoietin (IL-3/Epo) fusion protein have been constructed, expressed, and tested for biological activity. These fusion proteins were previously shown to be active on erythroid progenitors (BFU-E) from unseparated human bone marrow. We evaluated if these fusion proteins could stimulate erythroid and multipotential progenitor cells directly at the single-cell level. Two IL-3/Epo fusion proteins containing short (SL-3E, two amino acids) and long (LL-3E, 23 amino acids) linker sequences as well as a short linker Epo/IL-3 sequence (SL-E3, three amino acids) were tested. Highly enriched CD34 or BFU-E enriched CD34 CD33- cells from human umbilical cord blood or CD34 HLA-DR+CD33- cells from normal adult bone marrow were sorted as single cells into single wells. The combination of Epo plus IL-3 synergized to enhance the proliferation of BFU-E and multipotential progenitors (CFU-GEMM) in comparison to the individual effects of these cytokines. The three fusion proteins also enhanced proliferation of BFU-E and CFU-GEMM at the single-cell level and were at least as active as the combination of Epo and IL-3, demonstrating that IL-3/Epo fusion proteins directly stimulate proliferation of BFU-E and CFU-GEMM and that biological activity of IL-3 and Epo in vitro can be maintained when these proteins are fused. The activity of the combination of Epo and IL-3 or the fusion proteins was partially neutralized by preincubation with monoclonal antibodies to either Epo or IL-3 and was neutralized by greater than 90% by the combination of both antibodies, suggesting that the Epo and IL-3 components of the fusion proteins were both involved in the enhancing activity of these proteins. Additionally, use of monoclonal antibody to the human Epo receptor completely blocked the stimulating/enhancing activity of Epo alone, Epo plus IL-3, or the fusion proteins for stimulation of colony formation by BFU-E and CFU-GEMM but not for granulocyte-macrophage progenitors (CFU-GM), suggesting that the enhancing effects of the fusion proteins are most likely mediated, at least in part, by the Epo receptor.

Bone Marrow Cells↗

Hydroxyurea increases fetal hemoglobin in cultured erythroid cells derived from normal individuals and patients with sickle cell anemia or beta-thalassemia.

Hydroxyurea (HU), an inhibitor of DNA synthesis, has been shown to increase fetal hemoglobin (HbF) levels in patients with sickle cell anemia and in some patients with beta-thalassemia. However, until now there have not been good in vitro model systems that simulate this effect for study of the molecular and cellular mechanism(s) involved in perturbing the normal ontogeny of the globin genes. We analyzed the cellular effects of HU using a two-phase liquid culture procedure (Fibach et al: Blood 73:100, 1989) in which human peripheral blood-derived progenitor cells undergo proliferation and differentiation. HU was found to have multiple effects on these cultured cells: (1) an increase in the proportion of HbF produced; (2) a decrease in cell number due to inhibition of cell proliferation; (3) an increase in hemoglobin content per cell (mean corpuscular hemoglobin [MCH]); and (4) an increase in cell size (mean corpuscular volume). The extent of these effects was related to the HU dose and time of addition. When added to cell cultures from normal individuals, 4 days following their exposure to erythropoietin (EPO), 100 mumol/L HU caused a 1.3- to 3.5-fold increase in the proportion of HbF, from 0.4% to 5.2% (mean 1.6) in untreated to 1.5% to 8.2% (mean 3.1) in HU-treated cultures and a 45% +/- 10% increase in MCH but only a 25% +/- 7% decrease in cell number on day 13. Cultures of cells derived from five patients with sickle cell anemia have shown a twofold to fivefold increase in the percentage of Hb F following addition of HU while four patients with beta-thalassemia showed a 1.3- to 6.2-fold increase. We believe that this primary cell culture procedure should prove useful in studying the cellular and molecular mechanisms of pharmacologic induction of HbF and might provide a valuable predictive assay system for evaluation of the response of individual patients with hemoglobinopathies to HU and similar agents.

Anemia, Sickle Cell↗

Recombinant human mast cell growth factor supports erythroid colony formation in polycythemia vera in the presence and absence of erythropoietin and serum.

The effect of mast cell growth factor (MGF) was studied on erythropoietin (Epo)-dependent and Epo-independent ("spontaneous") erythroid colony formation in patients with polycythemia vera (PV). MGF stimulated both Epo-dependent and Epo-independent erythroid colony formation from PV peripheral blood progenitor cells in vitro at a dose similar to normal erythroid progenitor. In addition, evidence was obtained that the stimulating effect of MGF was a direct effect on the erythroid progenitor and independent of serum. Antibodies against interleukin-1 (IL-1), IL-3, granulocyte-macrophage colony-stimulating factor (GM-CSF), and Epo could not abolish the enhancing effect of MGF. This was also supported by the finding that sorted CD34+ cells could be stimulated by MGF in the presence and absence of Epo. Finally, it was demonstrated that the spontaneous erythroid colony formation could not be ascribed to spontaneous release of MGF in the culture medium since anti-MGF did not affect the colony numbers. In conclusion, MGF has a direct stimulatory effect, independent of serum, on both Epo-dependent and Epo-independent erythroid colony formation in PV.

Antibodies↗

In vitro sensitivity of human erythroid progenitors to hemopoietic growth factors: studies on primary and secondary polycythemia.

BACKGROUND: Primary proliferative polycythemia is a clonal disease characterized by excessive hemopoiesis and associated with a lower than normal erythropoietin plasma level; in vitro colony studies may reveal increased sensitivity of the abnormal clone to hemopoietic growth factors. MATERIALS AND METHODS: We studied the in vitro formation of erythroid colonies (BFU-E derived clone) in cultures set up with a serum-free medium and containing Epo, interleukin 3 (IL-3) and stem cell factor (SCF), in various combinations. The clonogenic test was performed by plating non adherent mononuclear cells from the peripheral blood of normal subjects and from patients with PPP and secondary polycythemia (SP). RESULTS: SCF is a major amplifier of erythroid colony growth, in the presence of Epo; in cultures from PPP patients, however, the presence of SCF, in addition to Epo, enhances colony formation at about the same rate as in cultures from normal subjects. When SCF is omitted, the presence of even modest amounts of Epo and IL-3 is sufficient to obtain a statistically significant difference between colony formation from PPP patients on the one side, and SP patients and normal subjects on the other. CONCLUSIONS: Our results show that in vitro culture studies may contribute an additional diagnostic criterion for distinguishing between PPP and SP in uncertain cases. It is also possible that hypersensitivity to erythropoietic factors may play a role in the pathogenetic mechanism of primary proliferative polycythemia.

Adult↗

[Erythroid stem (CFU-E) cells and erythropoietin levels in certain hematological diseases].

The aim of this paper is the trial of explanation of anaemia in certain immunological disorders. This approach consists measure of the Epo serum level and clot culture of bone marrow CFU-E cells. In the plasmocytoma patients inversely proportional dependence between number of CFU-E colonies and percentage of plasmocytes in bone marrow was observed. In low grade lymphoma patients the number of CFU-E colonies in vitro was greater than the standard value. Non proportional increase of Epo serum concentration to the level of anaemia in patients with acute leukemia was observed.

Blood Cell Count↗

The inhibitory action of aluminum on mouse bone marrow cell growth: evidence for an erythropoietin- and transferrin-mediated mechanism.

Aluminum (Al) has been associated with anemia in chronic renal failure patients under hemodialysis as well as in Al-overloaded animals. In an attempt to elucidate further the mechanism of Al toxicity we have investigated the effect of this ion on erythropoiesis in vitro. Mouse bone marrow cells were stimulated in vitro with erythropoietin (Epo) in the presence of Al3+ ion and erythroid colony-forming units were then determined. Results of this study indicate that Al compounds (chloride and citrate) at concentrations as low as 0.37 mumol Al/l inhibit erythropoiesis in vitro through a mechanism dependent upon the availability of transferrin to bind to aluminum. This process cannot be reversed by increasing Epo doses. This inhibition only occurs in the presence of Epo at early stages during the interaction of the hormone with its target cell.

Aluminum↗

Effects of activin A/erythroid differentiation factor on erythroid and megakaryocytic differentiations of mouse erythroleukemia (Friend) cells: evidence for two distinct modes of cell response.

To further characterize activin A/erythroid differentiation factor (EDF) action on hematopoietic cell differentiation, we examined the effects of activin A/EDF on megakaryocytic and erythroid differentiation by determining acetylcholinesterase (AchE) activity and hemoglobin production in the mouse erythroleukemia (MEL) cell line F55. Activin A/EDF induced AchE activity of F55 cells in a dose-dependent manner. Erythroid differentiation of F55 cells, which was characterized by an increase in dianisidine-positive cells, was also induced by activin A/EDF. The effect of activin A/EDF on hemoglobin synthesis appeared more slowly compared with the effect on AchE activity. Erythroid differentiation induced by activin A/EDF was affected by the initial cell density, but AchE activity was not. Sodium orthovanadate, a tyrosine phosphatase inhibitor, markedly inhibited activin A/EDF-induced erythroid differentiation but not activin A/EDF-induced AchE activity. Other erythroid differentiation inducers, sodium butyrate and butyrylcholine chloride, mildly increased AchE activity in F55 cells, but N,N'-hexamethylene-bis-acetamide (HMBA), dimethyl sulfoxide (DMSO), and genistein did not. Dexamethasone inhibited HMBA-induced erythroid differentiation but did not affect activin A/EDF or sodium butyrate action. These results suggest that F55 cells potentially can differentiate into cells of a megakaryocytic lineage in addition to an erythroid lineage, and that activin A/EDF further potentiates the cell differentiation of this cell line. In addition, our results suggest that the mode of activin A/EDF effects on megakaryocytic differentiation is distinct from that on erythroid differentiation.

Acetylcholinesterase↗

Inducible expression of a neo gene integrated into the human alpha-globin gene cluster.

We replaced the 3' flanking region of the human alpha 1-globin gene that binds in vitro the specific transcription factors GATA-1 and AP1/NF-E2, by a neo marker gene using homologous recombination in a MEL (mouse erythroleukemia line) hybrid cell line harbouring a single human chromosome 16. Using an improved method of the neo-positive and HSV-tk negative selection, one correctly targeted clone was obtained out of 164 clones analyzed. In contrast to non-targeted clones, the expression of teh neo gene in the targeted clone acquired the erythroid differentiation-dependent inducibility normally characteristic of the alpha-globin genes. No difference was observed in the expression of the human zeta, alpha 2, alpha 1, or theta-globin genes before and after induction of differentiation between the targeted clone and parental cells. These results indicate that, at least in the experimental system used, the 3' flanking region of the human alpha 1-globin gene can be replaced by an exogenous non-erythroid gene without affecting the regulation of the globin genes contained in the alpha-globin cluster.

Cell Differentiation↗

An endogenous signal triggering erythroid differentiation: identification as thyroid hormone.

The identification of thyroid hormone as an endogenous signal for erythroid differentiation began with our studies on the spontaneously differentiating murine erythroleukemia clone 3-1. We observed that the spontaneous differentiation frequency was dependent on a heat stable factor present in fetal calf serum or calf bone marrow. We also noted that the bone marrow extract stimulated erythroid colony-forming units in mouse bone marrow cells, suggesting the relevance of this factor in normal erythroid differentiation. The bone marrow extract did not supplant the requirement of erythropoietin but was synergistic. Purification of the bone marrow extract indicated that the differentiation-inducing activity for clone 3-1 cells cochromatographed with a low-molecular-weight, UV (280 nm)-absorbing component(s). These observations and previous reports identifying the avian erythroblastosis virus oncogene v-erbA as a mutated thyroid hormone receptor which blocked erythroid differentiation led us to test thyroid hormone in our assay. Both triiodothyronine and thyroxine were highly active, and the active constituents in the chromatographically purified fraction were identified as triiodothyronine and thyroxine. Although thyroid hormone action has been associated with both in vivo and in vitro erythroid differentiation, its role has been often relegated to a secondary status. We suggest that thyroid hormone is required for the commitment of erythroid cells to terminal differentiation.

Animals↗

Hematologic effects of recombinant human interleukin-6 in dogs exposed to a total-body radiation dose of 2.4 Gy.

The hematologic effects of recombinant human interleukin-6 (rhIL-6) were studied in dogs exposed to a total-body irradiation (TBI) of 2.4 Gy. IL-6 was administered over a period of 14 days at a daily dose of 18 micrograms/kg by single subcutaneous injection. Treatment was started 1 day after TBI. The data obtained for the different hematologic parameters of the irradiated IL-6-treated dogs were compared with the data obtained from dogs who received TBI of 2.4 Gy and were treated with the carrier (control). No clear influence of IL-6 treatment on the pattern of recovery of lymphocytes could be detected in comparison to the irradiated control animals. The thrombocyte counts in the period from day 1 to 16 after TBI were similar for both groups of dogs, showing a sharp decrease in counts between days 6 and 12 with a stabilization thereafter at approximately 30 x 10(3)/microL. In three of the four IL-6-treated dogs, however, thrombocyte counts increased at day 18 after the beginning of treatment. This increase occurred 7 days earlier than in the controls. In two of the three dogs showing an accelerated recovery of platelet counts, however, treatment with IL-6 caused a strong decrease in the erythrocyte counts associated with a prolonged depression in reticulocyte concentration. There was no influence on the recovery of blood granulocytes. In one of the animals responding with an accelerated thrombocyte recovery, IL-6 had no adverse effect on erythropoiesis. However, IL-6 forced the recovery of blood granulocytes in the period beyond day 10 after TBI. Another animal showed no influence of IL-6 on thrombocyte recovery but a strong depressive effect on erythrocyte and reticulocyte counts. The results show that for standardized conditions of radiation-induced bone marrow damage, the pattern of response to IL-6 in different hematopoietic lineages may show considerable variations between individuals, in contrast to what has been observed in irradiated animals treated with granulocyte-macrophage or granulocyte colony-stimulating factor (GM- or G-CSF).

Animals↗

Anti-erythropoietin (EPO) receptor monoclonal antibodies distinguish EPO-dependent and EPO-independent erythroid progenitors in polycythemia vera.

Erythroid progenitor cells isolated from patients with polycythemia vera (PV) proliferate and differentiate in methylcellulose in the absence of exogenous erythropoietin (EPO). To investigate the potential role of the erythropoietin receptor (EPO-R) in the pathogenesis of PV, we cultured bone marrow-derived or peripheral blood-derived erythroid progenitors in the presence of neutralizing monoclonal antibodies (MoAbs) specific for EPO or EPO-R. Mononuclear cells were obtained from 9 healthy adults and 9 PV patients by Ficoll-Hypaque gradients and cultured with or without EPO in methylcellulose for 12 days under standard or serum-free conditions. Neutralizing anti-EPO and anti-EPO-R MoAbs, added to cultures at day 0, caused dose-dependent growth inhibition of all normal burst-forming units-erythroid (BFU-E) derived from health adult controls. The MoAbs had no effect on the growth of nonerythroid progenitor cells under the same culture conditions. In contrast, neutralizing antibodies distinguished two classes of BFU-E derived from PV patients. Class I BFU-E from PV patients were EPO-dependent. These progenitors, like those derived from healthy adults, had normal EPO dose-dependent growth characteristics and showed a normal period of EPO requirement in vitro that extended 6 days after the initiation of culture. These results indicate that EPO exerts its critical effect early during erythroid differentiation; the addition of neutralizing antibodies to normal progenitors after 6 days had no effect on the subsequent size or maturation of the colonies. Class II BFU-E from PV patients were EPO-independent. They proliferated and differentiated even in the presence of high concentrations of neutralizing anti-EPO or anti-EPO-R MoAbs. We conclude that the class II BFU-E from PV patients are independent of free EPO.

Antibodies, Monoclonal↗