Search PubMed⌕ Search

Biomedical subjects

D Metcalf

Publications and source records attributed to D Metcalf.

At least 163 records · Page 9Linked to original sources
PubMed1992

Fatal polycythemia induced in mice by dysregulated erythropoietin production by hematopoietic cells.

C57BL/6J murine bone marrow cells, infected with a retroviral vector (MP Zen) carrying a monkey erythropoietin cDNA, were transplanted into lethally irradiated syngeneic recipients to study the effect of erythropoietin production by hemopoietic cells. High levels of erythropoietin were recorded in the plasma (median value: 1.2 u/ml) and in media conditioned by peritoneal, spleen, and bone marrow cells from recipient mice. In transplanted mice, the hematocrit was elevated (90 +/- 5%) and the mice died at a mean of 71 days after transplantation. In the blood, platelet counts were usually low and nucleated blood cells slightly elevated. Spleen weight increased 5-fold and bone marrow cellularity decreased slightly. There was a 9.9-fold increase in erythroblast numbers, a 2-fold reduction of lymphocytes, and no variation of the myeloid cells when the total cellularity of bone marrow, spleen, peripheral blood, and peritoneal cells were considered. Calculation of the total numbers of progenitor cells in these organs revealed a 18-fold increase in erythroid colony-forming units (CFU-E) but no significant variation of the erythroid burst-forming units (BFU-E), and myeloid progenitor cell numbers. A variable proportion of CFU-E, (12% or 24% in bone marrow or spleen, respectively) was able to proliferate in unstimulated cultures. Erythropoietic amplification occurred in the spleen and there was a redistribution of the BFU-E and myeloid cells from the bone marrow to the spleen. No significant extramedullary erythropoiesis was seen. This study emphasizes the erythroid specificity of erythropoietin and shows that elevated dysregulated erythropoietin production by hemopoietic cells leads to a fatal polycythemia without erythroid neoplastic transformation.

Animals↗
PubMed1992

Role of hemopoietic growth factors in the development and suppression of myeloid leukemia.

Myeloid leukemia development requires the acquisition by a cell of two abnormalities: an abnormal capacity for self-replication; and a capacity for autocrine stimulation, usually involving the known growth factors for granulocyte-macrophage cells. Curiously, in human leukemia, this does not usually result in autonomous growth when assessed in clonal in vitro cultures. Depending on gene programming, in particular in human or murine myeloid leukemias, the hemopoietic growth factors can also suppress the leukemic population by inhibiting the capacity of the leukemic stem cells for self-generation. The regulator showing the highest suppressive activity varies from leukemia to leukemia, with G-CSF. GM-CSF, IL-6, or leukemia-inhibitory factor (LIF) all having high activity on appropriate target cells. Combinations of these regulators are more effective than single agents alone. Analyses of human HL60, U937 and murine M1 leukemic models indicate that the development of morphological maturation in the leukemic cells is not a necessary feature of stem-cell suppression. LIF has an anomalous action on stem-cell self-generation, being highly effective in the suppression of certain myeloid leukemic cell lines, but being necessary to maintain self-generation in normal embryonic cells. This suggests the existence of a common control medium governing self-generation decisions in cells of different lineages, but that the outcome of the decision is determined by the differentiation program operating in different cells. The colony-stimulating factors are being used in combination with chemotherapy in the treatment of patients with acute myeloid leukemia, but the above principles require caution in certain situations.

Granulocyte-Macrophage Colony-Stimulating Factor↗
PubMed1991

Lineage commitment of hemopoietic progenitor cells in developing blast cell colonies: influence of colony-stimulating factors.

In clonal cultures of normal mouse marrow cells, combination of granulocyte, granulocyte-macrophage, or multipotential colony-stimulating factor (G-CSF, GM-CSF, or multi-CSF, respectively) with stem cell factor (SCF) did not alter the number of blast colonies stimulated to develop compared with SCF alone but induced an up to 25-fold increase in their mean cell content and an up to 6-fold increase in their mean progenitor cell content. Costimulation of blast colony formation by SCF plus G-CSF did not change the relative frequency of progenitor cells of different types within the colonies compared with colonies stimulated by SCF alone. However, combination of GM-CSF or multi-CSF with SCF significantly increased the relative frequency of granulocytic progenitors and, for multi-CSF, also of eosinophil progenitor cells. These changes in the relative frequencies of progenitor cells committed to the various lineages support the hypothesis that hemopoietic regulators have some ability to induce selective lineage commitment in the progeny of multipotential cells.

Animals↗
PubMed1991

Selective up-regulation of macrophage function in granulocyte-macrophage colony-stimulating factor transgenic mice.

Peritoneal and pleural cells from mice transgenic for GM-CSF were studied with regard to their phenotype and functional capacity, and compared with cells from normal littermates. Transgenic mice showed markedly elevated peritoneal and pleural cell counts compared with littermates, and a significantly higher proportion of cells in the transgenic populations were macrophage in phenotype. Transgenic macrophages were larger than the littermate cells, showing abundant foamy cytoplasm and enhanced spreading on plastic. Analysis by flow cytometry showed a more than sixfold increased expression of the macrophage activation markers MAC-2 and MAC-3, but not other markers, on transgenic macrophages. Superoxide production was measured in whole cell populations, both in their basal state and in response to particulate (zymosan) and soluble (PMA) stimuli. Both basal and stimulated superoxide production were markedly elevated in transgenic mice of 12 wk of age, with the largest differences seen in response to PMA. In younger mice, however, only PMA-stimulated superoxide production was significantly greater in transgenic macrophages than in littermate cells and levels of superoxide were generally lower than those seen in 12-wk-old mice. These findings suggest that the enhanced functional capacity of transgenic cells is a maturation-dependent event. In contrast to these findings, drug-dependent cytotoxicity assays performed on cells from 12-wk-old mice revealed no significant differences in killing capacity between the two mouse strains. Taken together these data indicate a selective rather than uniform functional up-regulation in transgenic macrophages compared with their littermates, with a time scale suggestive of a maturational rather than activation process. These findings may provide an indication of the functional macrophage phenotype resulting from long term exposure to GM-CSF in vivo, and help to explain the macrophage-associated pathology seen in GM-CSF-transgenic mice.

Animals↗
PubMed1991

Control of granulocytes and macrophages: molecular, cellular, and clinical aspects.

The production and functional activity of two important white blood cells, the granulocytes and macrophages, are regulated mainly by a group of glycoprotein colony-stimulating factors. The colony-stimulating factors have been mass-produced with recombinant technology and are now proving of value in preventing or suppressing infections in a variety of individuals with subnormal or defective formation of blood cells.

Acquired Immunodeficiency Syndrome↗
PubMed1991

The Florey Lecture, 1991. The colony-stimulating factors: discovery to clinical use.

The four colony-stimulating factors, GM-GSF, G-CSF, M-CSF and Multi-CSF, are specific glycoproteins with a likely common ancestral origin which interact to regulate the production, maturation and function of granulocytes and monocyte-macrophages. Each has been purified and produced in active recombinant form. Animal studies have shown the ability of injected CSF to increase the production and functional activity of granulocytes and macrophages in vivo and to enhance resistance to infections. These studies have led to the current extensive clinical use of CSFs to promote the formation and function of granulocytes and macrophages in a wide variety of disease situations in which there is an associated risk of serious infections. Although our knowledge of the control of haemopoiesis remains incomplete, the approaches used to develop the CSFs can be used to extend this knowledge, with the promise of the introduction into clinical medicine of additional effective therapeutic agents.

Amino Acid Sequence↗
PubMed1991

Direct proliferative actions of stem cell factor on murine bone marrow cells in vitro: effects of combination with colony-stimulating factors.

Stem cell factor (SCF), the ligand for the c-kit protooncogene product, was able to stimulate blast cell and granulocytic colony formation by precursors from normal murine bone marrow. The blast cell colonies contained a high content of progenitor cells able to form macrophage and/or granulocyte colonies. Clone transfer studies, the secondary culture of colony cells, and the culture of populations freed of accessory cells all indicated a direct proliferative action of SCF. SCF receptors were present in high numbers on blast cells and in lower numbers on immature granulocytic, monocytic, and eosinophilic cells. Combination of SCF with granulocyte, granulocyte-macrophage, or multipotential colony-stimulating factors, but not macrophage colony-stimulating factor, resulted in enhancement of colony size. Granulocyte colony-stimulating factor enhanced cell proliferation initiated by SCF, but not vice-versa, and resulted in a 10-fold increase in colony cell numbers and a 7-fold increase in progenitor cells in blast colonies. No evidence was obtained that SCF, alone or in combination with granulocyte colony-stimulating factor, could stimulate self-generation by blast colony-forming cells.

Animals↗
PubMed1991

Transgenic mice as models of hemopoiesis.

A useful approach to establishing the biologic actions, in vivo, of granulocyte-macrophage colony-stimulating factor (GM-CSF) is to assess the consequences of long-term elevation of the factor in transgenic mice. Two lines of transgenic GM-CSF mice were analyzed. The major abnormality exhibited was an elevation in the number of macrophages, eosinophils, and polymorphs in the peritoneal and pleural cavities. Disease states exhibited by the lines were dependent on the insertion site of the GM-CSF gene. These disease states seem best explained on the basis of CSF-mediated macrophage activation and may provide valuable clues as to the cause of comparable human diseases such as malignant histiocytosis, polymyositis, or rheumatoid arthritis.

Animals↗
PubMed1991

Leukemia inhibitory factor can potentiate murine megakaryocyte production in vitro.

Receptors for murine leukemia inhibitory factor (LIF) were demonstrated on immature and mature murine megakaryocytes. LIF alone had no effects in culture on the survival or proliferation of normal murine megakaryocytes or their precursors. However, combination of LIF with multipotential-colony-stimulating factor (Multi-CSF) (interleukin-3) enhanced the megakaryocyte colony formation able to be stimulated by Multi-CSF; the enhancement involved all types of megakaryocyte colony and resulted in the formation of increased numbers of megakaryocytes. These observations provide a possible basis for the observation that, when LIF is injected in vivo, elevations are observed in megakaryocyte numbers and platelet levels.

Animals↗
PubMed1991

Molecular cloning and chromosomal localization of the murine homolog of the human helix-loop-helix gene SCL.

The human SCL gene is a member of the family of genes that encode the helix-loop-helix (HLH) class of DNA-binding proteins. A murine SCL cDNA was isolated from a normal macrophage cDNA library by using HLH-specific oligonucleotides as hybridization probes. The coding region is 987 base pairs and encodes a predicted protein of 34 kDa. The nucleotide sequence of the coding region shows 88% identity to the human SCL gene, and the amino acid sequence is 94% identical. The HLH motif and upstream hydrophilic region are entirely conserved in the murine and human proteins. The identity between the mouse and human sequences was less marked in the 5' and 3' untranslated regions. Two murine SCL transcripts that differ in the 3' noncoding region have been detected in fetal liver and various cell lines. Variation was also observed in the 5' untranslated region. Interestingly, immediately downstream of the protein-termination codon, both the human SCL sequence and the murine homolog share an E-box element--the suggested target site for DNA binding of HLH proteins. The murine SCL homolog was mapped to the central part of chromosome 4.

Alleles↗
PubMed1991

Distribution and comparison of receptors for leukemia inhibitory factor on murine hemopoietic and hepatic cells.

Leukemia inhibitory factor (LIF) is a glycoprotein that induces the differentiation of the monocytic leukemia cell line M1 but suppresses the differentiation of totipotent embryonic stem cells. In an attempt to define the normal cellular targets for LIF, the distribution of LIF receptors within hemopoietic and hepatic tissue was analyzed by binding cells with radioiodinated LIF (125I-LIF) and subsequently carrying out autoradiography. Autoradiography demonstrated that in each hemopoietic tissue examined cells of monocyte/macrophage lineage were the primary cell type labeled with 125I-LIF. Moreover, both fetal and adult parenchymal hepatocytes displayed higher levels of labeling than either monocytes or macrophages. The number of receptors per positive cell varied from 150 for bone marrow monocytes to 2,000 for adult hepatocytes. In each case, however, binding was of high affinity, with an apparent KD of 34-100 pM, and binding was specific, since labeling was competed for by unlabeled LIF but not a range of other structurally unrelated growth and differentiation factors. It is suggested that LIF may play a role in regulating macrophage function and hepatic acute phase protein synthesis in response to infection.

Animals↗
PubMed1991

Clearance and fate of leukemia-inhibitory factor (LIF) after injection into mice.

Leukemia-inhibitory factor (LIF) elicits effects on a broad range of cell types, including cells of the monocytic and megakaryocytic series, embryonal stem cells, hepatocytes, adipocytes, and osteoblasts. Native and recombinant LIF, injected intravenously into adult mice, had an initial half-life of 6-8 min and a more prolonged second clearance phase. Clearance of 125I-LIF from the circulation was paralleled by a rapid accumulation in the kidneys, liver, lungs, and spleen and a more gradual accumulation in the thyroid gland. Labeling of the renal glomerular tufts, parenchymal hepatocytes, splenic red pulp, alveolar pneumocytes, and thyroid follicular cells as well as of megakaryocytes and osteoblasts in the bone cavities, placental trophoblasts, and cells of the choroid plexus was demonstrable autoradiographically. The appearance of a large amount of nonprecipitable 125I in the urine suggested that the kidneys were the major route of LIF clearance from the body.

Animals↗
PubMed1991

The leukemia inhibitory factor (LIF).

Leukemia inhibitory factor (LIF) is a glycoprotein able to enforce differentiation and/or suppress clonogenic self-renewal in a number of myeloid leukemic cell lines. When acting on normal embryonic stem cells, it has the opposite action of preventing differentiation commitment. LIF is not a proliferative factor when acting alone on normal hemopoietic cells, but can potentiate the action of interleukin 3 on blast cell and megakaryocyte precursors. When injected in vivo, LIF stimulates rises in megakaryocyte numbers and platelet levels. LIF also exhibits striking functional effects on a wide range of other cells including hepatic parenchymal cells, neurones, adipocytes, osteoblasts and gonadal cells. The polyfunctionality of LIF suggests strongly that it is normally intended to be produced locally and act as a local regulator. Despite its wide range of actions, LIF remains a promising candidate for clinical use in thrombocytopenia and myeloid leukemia.

Animals↗
PubMed1991

Segmentation of brain CT images using the concept of region growing.

A method is described for extracting and isolating cerebrospinal fluid and tissue areas of brain images obtained with computed tomography. The classification of the pixels into components is based on region growing and nearest neighbor principles. To aid the performance of this method, the algorithm utilizes a priori information on the anatomic composition of the brain, and reduces the 'cupping effect' in the CT image that is attributed to beam hardening artifacts. In order to avoid subjectivity, the performance of the algorithm was tested by superimposing five computer-simulated circular lesions on different areas of the original CT scans, 8 mm thick. These images were taken at different levels in the brain, thereby accommodating different anatomy as well as the apical artifact of CT scanning. In this exploratory investigation, the false negative error of segmentation for lesions having diameter of 20 pixels was found in the order of 25% at an estimated partial volume (PV) effect of 50% that decrease further to about 5% for a PV of 80%. At that point the false positive error becomes the dominant error in the analysis.

Algorithms↗
PubMed1991

Application of automated MRI volumetric measurement techniques to the ventricular system in schizophrenics and normal controls.

As an initial approach to computer-automated segmentation of cerebral spinal fluid (CSF) vs. brain parenchyma in MR scans, and the transformation of these data sets into volumetric information and 3D display, we examined the ventricular system in a sample of ten chronic schizophrenics with primarily positive symptoms and 12 normal subjects. While no significant differences were noted between groups on volumetric measures of ventricular brain ratio or lateral ventricle size, normals showed a pattern of left greater than right lateral ventricular volume asymmetry not present in the schizophrenics. Within the schizophrenic group, departure from the normal left greater than right pattern was highly correlated with thought disorder.

Adult↗
PubMed1991

Murine erythroid cell lines derived with c-myc retroviruses respond to leukemia-inhibitory factor, erythropoietin, and interleukin 3.

The transforming potential of the c-myc gene is shown here, for the first time, to include murine erythroid cells. Continuously growing cell lines were reproducibly generated by infection of day 13 CBA fetal liver cells with novel recombinant c-myc retroviruses. By cytostaining, most cells resembled early erythroblasts, but certain lines also contained significant numbers of hemoglobinized cells. RNA analysis revealed substantial expression of the genes encoding beta-globin and the erythroid-specific transcription factor GF-1. Although apparently immortal, the lines were not initially transplantable. Thus, constitutive myc expression in early erythroid cells can enhance their self-renewal capacity but is insufficient to fully transform them. The cell lines proliferated without the addition of exogenous factors, but their clonogenicity in semisolid medium was enhanced in the presence of erythropoietin, interleukin 3, and/or leukemia-inhibitory factor. In combination with either interleukin 3 or erythropoietin, leukemia-inhibitory factor also facilitated differentiation of certain lines. These results suggest that leukemia-inhibitory factor may have a previously unsuspected role in the regulation of erythropoiesis and could be considered as a possible therapeutic agent for the clinical management of erythroleukemia.

Animals↗