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Biomedical subjects

H E Broxmeyer

Publications and source records attributed to H E Broxmeyer.

At least 415 records · Page 23Linked to original sources
PubMed1978

Isolation of a granulocyte colony inhibitory factor derived from human polymorphonuclear neutrophils.

An inhibitory factor obtained from mature human granulocytes which suppresses granulocyte and monocyte-macrophage colony formation by an action on the endogenous colony stimulating factor-producing cells has been partially purified and characterized. The methods for purification consisted of a combination of ultracentrifugation, DEAE-Sephadex chromatography, SDS polyacrylamide gel electrophoresis and isoelectric focusing. The material had a molecular weight range of 102--128 000 and an isoelectric point between pH 6.2 and 6.4. The inhibitory factor was found to be heat stable and glycoprotein in nature.

Cell Differentiation↗
PubMed1978

Limitation of excessive myelopoiesis by the intrinsic modulation of macrophage-derived prostaglandin E.

The clonal proliferation of the committed granulocyte-macrophage stem cell is controlled by a balance between mutually opposing factors, colony stimulating factor and prostaglandin E, both of monocyte-macrophage derivation. Increases beyond a critical concentration of colony stimulating factor within the local milieu of the mononuclear phagocyte induces the coincident elaboration of prostaglandin E, a self-regulated response which serves to limit the unopposed humoral stimulation of myelopoiesis.

Animals↗
PubMed1978

Immunologic functions and in vitro activation of cultured macrophage tumor lines.

Five murine monocyte of macrophage tumor lines adapted to culture were characterized for differentiated properties. They ingested zymosan and latex beads, bore receptors for immunoglobulin and complement, synthesized lysozyme (most of which was secreted), and produced granulocyte colony-stimulating activity, either spontaneously or inducibly. Some of the lines also mediated phagocytosis and exocytosis of red blood cells (RBC) and lysis of tumor targets, dependent on the presence of specific antitarget sera. All the lines were growth inhibited by zymosan and Mycobacterium bovis BCG, but not by latex beads. Other macrophage-activating agents, dextran sulfate and lipopolysaccharide (LPS), as well as tuberculin purified protein derivative (PPD), inhibited most of the lines. Except for Fc and C receptors, most of the above properties were not found with other types of hematopoietic tumors in culture. In attempts to activate the macrophage lines in vitro to the "angry" state, we found that preincubation with concentrations of LPS and PPD cytostatic to the cells stimulated antibody-dependent RBC lysis, but not antibody-independent or tumor cytolysis. A classification of monocyte-related tumors and normal cells is proposed based on functional activities and differential sensitivity to immunostimulating agents.

Animals↗
PubMed1978

Induction of myeloid colony-stimulating activity in murine monocyte tumor cell lines by macrophage activators and in a T-cell line by concanavalin A.

Certain fibrosarcoma lines in culture and the WEHI-3 myelomonocytic leukemia cell line have previously been shown to secrete myeloid colony-stimulating activity (CSA) spontaneously. We describe here other hematopoietic tumor cell lines in which CSA is either produced constitutively or inducible by immunostimulators. CSA production in macrophage and monocyte tumor lines is induced by lipopolysaccharide, zymosan, Mycobacterium strain Bacillus Calmette-Guérin, tuberculin purified protein-derivative preparation from mycobacteria, and dextran sulfate. Myeloma, mastocytoma, and T-lymphoma lines do not produce CSA with or without these agents. In contrast, the T-lymphocyte mitogen concanavalin A (but not phytohemagglutinin) induces CSA synthesis in one of seven T-lymphomas tested. In most cases induction of CSA is correlated with conditions of cell growth inhibition by the immunomodulators. However, other drugs that cause cytostasis or cytotoxicity do not lead to CSA production. Leukemic cells thus may retain sensitivity to normal regulatory events with resultant effects on host hematopoietic cell functions.

Animals↗
PubMed1978

Inhibition in vivo of mouse granulopoiesis by cell-free activity derived from human polymorphonuclear neutrophils.

Cell-free extracts from human polymorphonuclear neutrophils (PMN) inhibited rebound granulopoiesis in the bone marrow and spleen of mice pretreated with cyclophosphamide to remove endogenous PMN. Absolute numbers of granulocytemonocyte progenitor cells (CFU-C) and net endogenous colony-stimulating activity (CSA) production were found to be increased 3 days after cyclophosphamide in the bone marrow and 6 days after in the spleen. Administration of PMN extract to the drug-treated mice prior to rebound granulopoiesis substantially decreased CSA production and CFU-C but not spleen B lymphocyte colony-forming cells. In addition, mice treated with PMN extract had decreased levels of CSA in serum and in conditioned medium of marrow-free bone, heart, and lung cultures. Inhibition was reversed by injection of bacterial lipopolysaccharide. Extracts from PMN of patients with chronic myelogenous leukemia, inactive in vitro, had no effect in vivo. These results demonstrate that inhibitory activity derived from PMN can control granulopoiesis in vivo.

Animals↗
PubMed1978

Corticosteroids block newly induced but not constitutive functions of macrophage cell lines: myeloid colony-stimulating activity production, latex phagocytosis, and antibody-dependent lysis of RBC and tumor targets.

Hydrocortisone at 10(-4) M added to macrophage cell lines had no effect on the following activities: J774.1 antibody-dependent phagocytosis and lysis of sheep RBC, RAW 264 antibody-dependent lysis of tumor targets, latex bead phagocytosis by PU5-1.8 cells, and constitutive production of myeloid colony-stimulating activity (CSA) by WEHI-3 cells. In contrast, 10(-4) M hydrocortisone completely inhibited LPS or PPD activation of the following functions: induced production of myeloid CSA, stimulation of latex phagocytosis and stimulation of antibody-dependent lysis of tumor targets by PU5-1.8 cell line, and stimulation of antibody-dependent lysis of RBC targets by RAW 264 cell line. Lower concentrations of hydrocortisone or dexamethasone at 10(-6) M inhibited the LPS induction of CSA from 55 to 90%. Growth inhibition of PU5-1.8 cells caused by 1 microgram/ml LPS was also partially reversed by corticosteroids. Thus, steroids block these newly induced but not constitutive functions of these macrophage cells in culture.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1977

Immunostimulators induce granulocyte/macrophage colony-stimulating activity and block proliferation in a monocyte tumor cell line.

Monocyte tumor cell line PU5-1.8 does not normally produce colony-stimulating activity (CSA) required by granulocyte and macrophage progenitors to proliferate and mature in agar. However, CSA is induced in the culture line by as little as 10 ng/ml endotoxic lipopolysaccharide (LPS), with maximum CSA production and release to the medium between 2 and 3 days of incubation. Derived lipid A, but not alkali-treated LPS, is also active. Induction requires RNA and protein synthesis, but is not blocked by mitomycin C or Colcemid. Other inducers of CSA include Mycobacterium Bacillus Calmette-Guérin, tuberculin protein preparation purified protein derivative, zymosan, and phorbol myristate. All inducing agents are specific inhibitors of the monocyte tumor cell proliferation in vitro. Latex beads, another macrophage-activating agent, are rapidly phagocytosed by PU5-1.8 cells, but neither inhibit growth nor induce CSA.

Animals↗
PubMed1977

Expression of Ia-like antigen molecules on human granulocytes during early phases of differentiation.

Human B lymphocyte antigens analogous to the murine Ia determinants were found on myeloblasts and promyelocytes but not on more mature granulocytes. This was apparent by fluorescent staining with both human alloantisera and rabbit antisera to the isolated Ia-like proteins. The cells of patients with chronic myelocytic leukemia showed this difference especially clearly. Separation of the myeloblasts and promyelocytes by multistep density gradient fractionation produced a marked enrichment of the positive cells. The remaining cells from higher density fractions were more-mature neutrophils that were essentially negative. In acute myeloid leukemia, in which myeloid cells early in differentiation predominate, the vast majority of cells were strongly positive. Similar results were obtained with normal bone marrow cells. Here also, only the early forms of the myeloid series separated by gradient centrifugation had Ia antigens. Evidence was also obtained for the presence of Ia determinants on cells with the appearance of early erythroid precursors. Support for the presence of the Ia determinants on granulocyte-macrophage committed stem cells was provided by the inhibition of granulocyte colony formation in agar cultures following preincubation of normal bone marrow with antiserum and complement. Cross absorptions with purified preparations of immature cells provided evidence for the close similarity of the antigenic determinants on both myeloblasts and B cells. A 28,000-37,000-dalton bimolecular complex obtained from myeloblast membranes contained the Ia determinants and was similar to that obtained from peripheral blood B cell membranes.

B-Lymphocytes↗
PubMed1977

Cell-free granulocyte colony inhibiting activity derived from human polymorphonuclear neutrophils.

Human PMN (non-adherent: density greater than 1.070 g/cm3) were used to prepare cell-free colony inhibiting activity (CIA). Intact PMN, PMN conditioned media (serum-free or serum-containing) and supernatant from immediately lysed PMN equally reduced the numbers of spontaneous bone marrow (density less than 1.070 g/cm3; depleted of PMN) colonies and clusters. Addition of CIA reproducibly reduced colony and/or cluster numbers by 35-70%. CIA was heat labile being inactivated at 37 degrees C. Daily additions of CIA reduced colony numbers even further; inhibition was dependent on [PMN] used to prepare CIA and on the potency of stimulation produced by colony stimulating cells (CSC). PMN from some patients with leukemia were inactive as inhibitors and did not condition media with CIA. All experiments were thereafter controlled by parallel assessment of supernatant of lysed PMN not demonstrating any CIA. CIA appears to be not only stored in PMN but released from viable, as well as degranulating PMN. Although not species-specific, CIA was specific in its nontoxic action on CSC; colony forming cells and cell-free colony stimulating activity were not affected. The influence of CIA on CSC could be reversed within 10 minutes, but not within 35 minutes of incubation. CIA may have possible physiological relevance.

Animals↗
PubMed1977

In vitro regulation of a mouse myelomonocytic leukemia line adapted to culture.

Regulation of the growth and production of granulocyte colony-stimulating activity (CSA) by WEHI-3 cells, a lysozyme-secreting mouse cell line adapted to culture, was investigated in vitro. WEHI-3 cloning efficiency is not enhanced by exogenously added CSA. However, WEHI-3 cloning efficiency in agar was suppressed by an activity in human polymorphonuclear neutrophil extract (colony-inhibiting activity) which inhibits endogenous WEHI-3 CSA production. The addition of increasing concentrations of WEHI-3- or L cell-conditioned medium containing CSA to CIA-depressed WEHI-3 agar cultures resulted in graded increases of cloning efficiency to that of the untreated sample. Testosterone, Deca-Durabolin, and bacterial lipopolysaccharide increased production of CSA by WEHI-3 cells and overcame colony-inhibiting activity-mediated suppression of CSA production, even when activating agents were added 1 day after the addition of colony-inhibiting activity. The activating agents had no direct stimulatory effect on normal mouse marrow colony-forming cells and did not enhance CSA activity. WEHI-3 cells respond to growth inhibitory and stimulatory activities and can serve as an in vitro model for studying the regulation of neoplastic cells.

Animals↗
PubMed1976

In vitro regulation of granulopoiesis in human leukemia: application of an assay for colony-inhibiting cells.

We describe two assays to detect the action of colony-inhibiting cells. In the first assay, we used a simple density separation technique to remove dense neutrophils (PMN) from suspensions of blood and of bone marrow cells prior to culture in semisolid agar. Conditions were arranged to ensure that control suspensions of unseparated cells and test suspensions of buoyant mononuclear cells differed only in their content of neutrophils. The control and test suspensions contained equal numbers of mononuclear cells (and granulocyte precursors). Colony (and cluster) formation was invariably enhanced in neutrophil-depleted cultures of normal cells. In the second assay, dense PMN, treated by an adherence separation procedure, were recovered, and the non-adherent dense PMN were added back to PMN-depleted cultures. A reproducible dose-related decrease in colony (and cluster) formation to basal levels resulted. The inhibitory effect was identical when the PMN were added directly to the culture (overlayer) or to the underlayer. In PMN-depleted cultures obtained from patients with leukemia and other hemopoietic disorders, neither colony nor cluster formation was enhanced, and sometimes it was reduced. When we compared the effect of adding patient and normal non-adherent PMN to target cultures of normal and patient PMN-depleted cells, some leukemic PMN were noninhibitory. Our results suggest that abnormalities of cellular interactions in vitro detected in the first assay may have more than one explanation, as shown when they are subjected to the closer scrutiny possible with the second assay.

Bone Marrow↗
PubMed1976

Relationship of colony-stimulating activity to apparent kill of human colony-forming cells by irradiation and hydroxyurea.

Suspensions of human bone marrow cells were subjected to 137Cs irradiation in vitro and then cultured in semisolid agar medium. Cultures of irradiated cells were stimulated with colony-stimulating activity (CSA) of different potencies, and it was found that the amount of stimulation applied to cultures influenced the apparent kill of colony-forming cells (CFC). It was also found that the effects of irradiation on colony formation were not confined to CFC kill since medium conditioned by cells during irradiation exhibited stimulatory and inhibitory properties after treatment by 600 and 1000 rads, respectively. Studies in which irradiated cells were pretreated with hydroxyurea indicated that CFC in the DNA synthetic phase of the cell cycle were particularly sensitive to low doses of irradiation. The proliferative capacity of CFC surviving 1000 rads was undiminished as judged by their ability to form large colonies. Estimates of CFC kill by hydroxyurea were also affected by the level of CSA.

Bone Marrow↗
PubMed1975

Control of granulopoiesis in man.III. inhibition of colony formation by dense leukocytes.

Cellular feeder layers, prepared from normal blood leukocytes, usually stimulate human marrow to form colonies. A significant increase in the stimulating activity of unseparated leukocyte feeder layers is brought about following the removal of dense leukocytes in a manner which avoids enrichment of any remaining cell type. Restoration of dense leukocytes to a dense leukocyte depleted leukocyte feeder layer results in the reduction of stimulating activity to that of an unseparated leukocyte feeder; however, addition of dense leukocytes to unseparated leukocyte feeder layers has no effect on the stimulatory activity, over the range of concentrations used in this study.

Bone Marrow Cells↗