Colony-stimulating Activity Production Research Articles

Chlorella vulgaris Modulates Immunomyelopoietic Activity and Enhances the Resistance of Tumor-Bearing Mice

We studied the effects of Chlorella vulgaris (CV) on the interaction between stromal and hematopoietic stem cells in normal and Ehrlich ascites tumor (EAT)-bearing mice. Long-term bone marrow culture (LTBMC), cytokine production, spleen mononuclear cells (SMC) proliferation (SCP), colony stimulating activity (CSA), and NK cells activity were evaluated. In tumor bearers, reduced capacity of stromal cell layer to support the growth and differentiation of granulocyte-macrophage progenitor cells (CFU-GM), concomitantly to decreased numbers of total nonadherent cells in LTBMC and reduced local production of IL-6 and IL-1α, were observed. Presence of the tumor has not altered the number of stromal adherent cells. CV treatment restored the ability of stromal cells from EAT-bearing mice to produce IL-6 and IL-1α, which was consistent with increased number of nonadherent cells and higher ability to display CFU-GM in vitro. EAT growth increased SCP, serum CSA, and IL-10 production and concurrently depressed NK cell activity and the secretion of IL-2, IFN-γ, and TNF-α. Treatment of tumor-bearing mice with CV augmented CSA, SMC proliferation, NK cell activity, and the production of IL-2, IFN-γ, and TNF-α, whereas IL-10 levels where reduced. Our results suggest that CV modulates immunehematopoietic cell activity and disengages tumor-induced suppression of these responses.

Production of colony-stimulating activity by a murine bone marrow stromal cell line on porous microcarriers

The murine bone marrow stromal cell line, LC2, cultivated on porous microcarriers, displayed cell density-dependent growth, which was alleviated by its own conditioned-medium. Volumetric production of the colony-stimulating activity in the microcarrier culture was many fold over that obtained from the T-flask culture, providing a means of scaling up for production.

Interaction of Murine Peritoneal Leukocytes and Mesothelial Cells: In Vitro Model System to Survey Cellular Events on Serosal Membranes during Inflammation

All serosal cavities including peritoneum are lined with a simple squamous mesothelium. Primary culture of murine mesothelial cells has been established to study their cellular interactions with peritoneal leukocytes. The mesothelial character was determined by the cytokeratin and vimentin expression. The mesothelial cells expressed ICAM-1 and CD44 molecules. The expression of ICAM-1, but not CD44, was significantly enhanced by the treatment with TNF-α (100 U/ml). We have also investigated possible influence of transforming growth factors, TGF-α (20 ng/ml) and TGF-β (2 ng/ml), and epidermal growth factor (20 ng/ml). These factors were not found to modulate ICAM-1 or CD44 expression in vitro. During coculture experiments unstimulated mesothelial cells were almost nonadherent for both resident and elicited peritoneal mononuclear leukocytes for several hours. TNF-α or EGF pretreatment of mesothelial cells greatly enhanced their adhesive affinity to peritoneal mononuclear leukocytes, while TGF-β pretreatment even reduced the low basal adhesion. Prolonged coculture for 3 weeks resulted in remarkable proliferation and differentiation of both resident and elicited monocytes/macrophages on the mesothelial surface. The stimulation of mesothelial cell culture with EGF resulted in the macrophage colony-stimulating activity (M-CSA) production. M-CSA was mainly due to M-CSF as confirmed with anti M-CSF monoclonal antibody; the residual M-CSA was not formed by GM-CSF. After several passages the mesothelial cells started to produce M-CSA spontaneously.

Induction of non-specific suppressor cells and myeloregulatory effects of an immunomodulatory azaspirane, SK&F 105685

Administration of the immunosuppressive agent, SK&F 105685, has demonstrated immunosuppressive activity in several animal models of autoimmunity such as adjuvant arthritis and experimental autoimmune encephalomyelitis. The mechanism of action of SK&F 105685 in these autoimmune disease models appears to be the induction of non-specific suppressor cells (SC) detected in the spleen and bone marrow of treated animals. In this study we have examined the kinetics of SC appearance in the spleen and bone marrow following treatment with 30 mg/kg/day, p.o., for 1–6 days. SC activity was apparent following a single dose and increased with successive treatments. Treatment with SK&F 105685 also resulted in significantly enhanced myelopoiesis as measured by a 128% increase in the frequency of bone marrow myeloid progenitors (CFU-GM). Mechanistic studies indicated that in vitro treatment of bone marrow stromal cell cultures with SK&F 105685 upregulated the production of colony stimulating activity (CSA) detectable in a rat CFU-GM assay. Further, in vitro studies revealed that the SC in the bone marrow or spleens of SK&F 105685-treated rats admixed with normal marrow cells inhibited CFU-GM formation at a six-fold less cell concentration than cells obtained from control rats. These in vitro results suggest that the SK&F 105685-induced myelopoiesis is regulated by the subsequent generation of SC.

Effects of aflatoxin B 1 on myelopoiesis in vitro

The short-term cultures of mouse myeloid progenitor cells for granulocytes and monocytes, granulocyte-monocyte colony-forming units (CFU-GM) (CFU-GM assay) and mouse long-term bone marrow culture (LTBMC) were used to investigate the hemopoietic suppression caused by aflatoxin B 1 (AFB 1). A dose-related suppression of granulopoiesis in short-term bone marrow cultures was seen when the cultures were treated with 10, 5, 1, 0.5 and 0.1 μg of AFB 1/ml. Two selected doses of AFB 1 (5 and 0.5 μg/ml) considered to be highly and slightly suppressive in CFU-GM assay exerted a strong suppression of myelopoiesis in LTBMC when applied long-term. Short-term (2 h) exposure of LTBMC to 5 μg of AFB 1/ml caused a small damage to the myelopoiesis detected in the non-adherent layer. Short-term exposure to 0.5 μg AFB 1/ml was without any effect on myelopoiesis in LTBMC. The production of colony-stimulating activity (CSA) by an adherent layer of LTBMC was decreased on the second and fifth day after the short-term exposure to both doses of AFB 1 and comparable with non-treated culture on the seventh day after the exposure. Presented results indicate that both short-term culture of CFU-GM and LTBMC can be used in the definition and the prediction of host toxicity of AFB 1 to hemopoiesis. However, comparing these two in vitro systems, the LTBMC appears to be more sensitive and discriminatory in an evaluation of hemopoietic toxicity.

Production of Colony-Stimulating Activity by Circulating Leukocytes from Patients with Chronic Myeloid Leukemia at Various Phases of the Disease

The production of colony-stimulating activity (CSA) by peripheral blood leukocytes (PBL) from 107 patients with chronic myeloid leukemia (CML), 58 at diagnosis, 34 during hematological remission, 33 in relapse from hematological control, 39 in accelerated phase, and 28 in blast phase, was measured in the double-layer agar culture system using normal nonadherent bone marrow cells as the source of CFU-GM. The median CSA levels in various stages decreased to 28-60% of control, whereas in hematological remission, there was a normalization of CSA. Five fractionation experiments using monocytes as feeder layers did not show CSA production in untreated CML. The leukemic PBL of 6 patients in blast crisis had no apparently inhibitory effect on colony formation stimulated by normal PBL. There was no correlation between the CSA of PBL and the number of CFU-GM in their bone marrow or peripheral blood at different disease states. Likewise, we failed to find a relationship between the PBL CSA levels and the total WBC counts or the differential counts in the peripheral blood as well as in the feeder layers. Our study indicated that circulating leukocytes from patients with CML at various phases except in hematological remission produce less CSA, which is not attributed to a low number of monocytes or the inhibitory effects of leukemic cells.

Hematopoietic and hematologic properties of bestatin in normal and cyclophosphamide myelosuppressed mice

Bestatin is a potent inhibitor of aminopeptidase, an enzyme found in abundance in the membrane of monocytes and macrophages. Following binding of bestatin to cells in the histiocytic lineage, the production of colony stimulating activity is upregulated (both in vitro and in vivo) with subsequent increases in hematopoietic and hematologic parameters. We report that the frequency and absolute numbers of CFU-GM as well as entry of CFU-GM into S phase (a measure of progenitor cell activity) is upregulated by treatment of animals with bestatin. This results in an increase in bone marrow cellularity in cyclophosphamide suppressed mice and an increase in the absolute neutrophil count in normal and suppressed mice. The therapeutic application of this hematopoietic modulator has been demonstrated in combination cyclophosphamide and bestatin therapeutic protocols. Because this indication for bestatin has only recently been recognized, few clinical studies have been undertaken utilizing appropriate surrogates of hematopoietic activity. However, preliminary clinical evidence of hematopoietic activity by this non-toxic dipeptide, as reviewed here, suggests that this may be an appropriate strategy for the treatment of myelosuppressed patients.

Interleukin-6 is not involved in the interleukin-1-induced production of colony-stimulating factors by human bone marrow stromal cells and fibroblasts

Interleukin-6 (IL-6) is a multifunctional cytokine that plays a role in regulation of hematopoiesis. Because IL-6 is coinduced with colony- stimulating factors (CSFs) by various cell types in response to stimulation with IL-1, we investigated whether IL-6 is involved in the IL-1-induced production of CSF by human bone marrow (BM) cells in long- term culture or human fibroblasts. We showed that IL-6 does not induce CSF production by these cells. Neither addition of exogenous IL-6 nor neutralization of endogenous production of IL-6 by an anti-IL-6 monoclonal antibody (MoAb) diminished the IL-1-induced colony- stimulating activity (CSA), indicating that IL-6 did not act synergistically with IL-1. Finally, IL-6 did not influence the kinetics of IL-1-induced CSA production by human fibroblasts. We conclude that IL-6, either alone or in combination with IL-1, does not induce CSF production by human BM stromal cells or fibroblasts.

Interleukin-6 Is Not Involved in the Interleukin-1-Induced Production of Colony-Stimulating Factors by Human Bone Marrow Stromal Cells and Fibroblasts

Interleukin-6 (IL-6) is a multifunctional cytokine that plays a role in regulation of hematopoiesis. Because IL-6 is coinduced with colony-stimulating factors (CSFs) by various cell types in response to stimulation with IL-1, we investigated whether IL-6 is involved in the IL-1-induced production of CSF by human bone marrow (BM) cells in long-term culture or human fibroblasts. We showed that IL-6 does not induce CSF production by these cells. Neither addition of exogenous IL-6 nor neutralization of endogenous production of IL-6 by an anti-IL-6 monoclonal antibody (MoAb) diminished the IL-1-induced colony-stimulating activity (CSA), indicating that IL-6 did not act synergistically with IL-1. Finally, IL-6 did not influence the kinetics of IL-1-induced CSA production by human fibroblasts. We conclude that IL-6, either alone or in combination with IL-1, does not induce CSF production by human BM stromal cells or fibroblasts.

Effect of antilymphocyte globulin (ALG) on bone marrow T/non-T cells from aplastic anaemia patients and normal controls.

The aim of this study was twofold: (a) to test the effect of antilymphocyte globulin (ALG) on bone marrow (BM) T/non-T cells, and (b) to look for a possible differential response of cells from severe aplastic anaemia (SAA) patients and controls. For this purpose bone marrow T/non-T cells from normal individuals (n = 7) or aplastic patients (SAA, n = 13) were kept in liquid culture with or without ALG. Supernatants were then tested for enhancement/suppression on colony forming unit, granulocyte-macrophage (CFU-GM) growth (in the presence of exogenous recombinant granulocyte-macrophage colony stimulating factor (rGM-CSF)), or for their ability to support CFU-GM growth (in the absence of exogenous rGM-CSF). Supernatants from SAA T cells suppressed CFU-GM growth of normal bone marrow cells in 5/12 patients (mean expected growth (EG) 71 +/- 16%), but not after incubation with ALG (mean 110 +/- 29% EG, P = 0.03). No inhibition could be obtained with the supernatants from untreated normal T cells. Significant enhancement was seen with ALG treated versus untreated SAA T cells (142 +/- 28% EG v. 105 +/- 61% EG, P = 0.01) and with ALG treated versus untreated SAA non-T cells (165 +/- 26% EG v. 105 +/- 23% EG, P = 0.01), but not in controls. Supernatants from SAA and control T/non-T cells were capable of promoting colony formation in the absence of rGM-CSF (colony-stimulating activity (CSA) production): 16 +/- 14% for SAA-T cells and 19 +/- 18% EG for non-T cells (100% = 30 ng rGM-CSF/ml). The addition of ALG increased CSA production in T cells to 37 +/- 23% EG (P = 0.04) and in non-T cells to 40 +/- 13% EG (P = 0.04). Similar results could be obtained in controls. (a) ALG interacts in vitro with bone marrow T and non-T cells from SAA patients, down-regulating the production of negative lymphokines and enhancing the release of haemopoietins; (b) the latter, but not the former effect, can be shown also with cells from normal controls. The two effects are not mutually exclusive, and are likely to provide maximal benefit in vivo.

Lactoferrin decreases monocyte-induced fibroblast production of myeloid colony-stimulating activity by suppressing monocyte release of interleukin-1

Lactoferrin (Lf) is a negative regulator of myelopoiesis which operates by suppressing the release from mononuclear phagocytes of GM colony- stimulating factor (GM-CSF) or monokines which can then induce the release of GM-CSA from accessory cells. In this study, endotoxin- depleted, purified iron-saturated human Lf was assessed for its effect on the production of interleukin-1 by cultured monocytes and their subsequent effect on colony-stimulating factor release from cultured fibroblasts. Monocytes were grown with or without Lf and Lf that had previously been incubated with monoclonal anti-Lf. The monocyte- conditioned medium was then either assayed for the presence of interleukin-1 (IL-1) with an enzyme-linked immunosorbent assay or for its ability to stimulate fibroblasts to release growth factors for CFU- GM, BFU-E, or CFU-Mix colonies. In the presence of Lf (10(-7) or 10(-8) mol/L), GM colony-stimulating activity (GM-CSA) was suppressed by 31% to 73%, whereas stimulating activities for BFU-E and CFU-mix colony formation were suppressed by 93% to 100%. Antibody to Lf completely abrogated the suppressive effects observed with Lf, whereas antibody to IL-1 ablated the induction by monocyte-conditioned medium of CSA release by fibroblasts. Lf at 10(-7) and 10(-8) mol/L also reduced IL-1 synthesis by cultured monocytes from 60% to 77%. The inhibitory effects of Lf were only observed when Lf was added before adherence of the monocytes for culture. If Lf was added at the time of adherence or after adherence, no suppression was observed. We conclude that the inhibition of GM-CSA production/release by Lf is mediated through inhibition of the synthesis/release of IL-1 by mononuclear phagocytes. This inhibition of IL-1 prevents accessory cells from producing and/or releasing GM-CSA.

Lactoferrin Decreases Monocyte-Induced Fibroblast Production of Myeloid Colony-Stimulating Activity by Suppressing Monocyte Release of Interleukin-l

Lactoferrin (Lf) is a negative regulator of myelopoiesis which operates by suppressing the release from mononuclear phagocytes of GM colony-stimulating factor (GM-CSF) or monokines which can then induce the release of GM-CSA from accessory cells. In this study, endotoxin-depleted, purified iron-saturated human Lf was assessed for its effect on the production of interleukin-1 by cultured monocytes and their subsequent effect on colony-stimulating factor release from cultured fibroblasts. Monocytes were grown with or without Lf and Lf that had previously been incubated with monoclonal anti-Lf. The monocyte-conditioned medium was then either assayed for the presence of interleukin-1 (IL-1) with an enzyme-linked immunosorbent assay or for its ability to stimulate fibroblasts to release growth factors for CFU-GM, BFU-E, or CFU-Mix colonies. In the presence of Lf (10(-7) or 10(-8) mol/L), GM colony-stimulating activity (GM-CSA) was suppressed by 31% to 73%, whereas stimulating activities for BFU-E and CFU-mix colony formation were suppressed by 93% to 100%. Antibody to Lf completely abrogated the suppressive effects observed with Lf, whereas antibody to IL-1 ablated the induction by monocyte-conditioned medium of CSA release by fibroblasts. Lf at 10(-7) and 10(-8) mol/L also reduced IL-1 synthesis by cultured monocytes from 60% to 77%. The inhibitory effects of Lf were only observed when Lf was added before adherence of the monocytes for culture. If Lf was added at the time of adherence or after adherence, no suppression was observed. We conclude that the inhibition of GM-CSA production/release by Lf is mediated through inhibition of the synthesis/release of IL-1 by mononuclear phagocytes. This inhibition of IL-1 prevents accessory cells from producing and/or releasing GM-CSA.

In vivo and in vitro production of haemopoietic colony-stimulating activity by murine cell lines of different origin: a frequent finding

The production of colony-stimulating factor (CSF) by murine transformed cells was investigated in 10 cell lines derived from spontaneous or chemically induced tumours and from cells transformed by SV 40 or Moloney-MSV; histologic types included carcinomas, sarcomas and melanoma. Nine of 10 supernatants contained CSF activity as judged by in vitro proliferation and differentiation of normal murine monocytic and granulocytic progenitors in agar cultures. Tumours induced with CSF-producing cells caused alterations of haemopoiesis which can include leukocytosis, granulocytosis and splenomegaly. Haemopoietic alterations were also evident in the absence of a local tumour in mice bearing large experimental lung metastases. Production of CSF seems to be a frequent finding among murine cell lines, and its biological and immunological consequences on host-tumour relationships should be taken into account.

Production of colony-stimulating activity by human natural killer cells: analysis of the conditions that influence the release and detection of colony-stimulating activity

Highly purified natural killer (NK) cell suspensions were tested for their capacity to release colony-stimulating activity (CSA) in vitro. NK cell suspensions comprised primarily CD16+ cells and were devoid of CD3+ T cells, CD15+ monocytes, and of B cells. CSA was detected in the NK cell supernatants and sustained the growth of myeloid colonies from both normal peripheral blood and bone marrow. CSA could be in part inhibited by pretreating NK cell culture supernatants with a specific goat anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) antiserum. The inhibition, however, was never complete, a finding that suggests that additional factors were responsible for CSA. Incubation of NK cells with K562 cells (an NK-sensitive target) or with normal bone marrow cells resulted in the appearance of a strong colony- inhibiting activity (CIA) in the culture supernatants. Such CIA was demonstrable in an experimental system where bone marrow or peripheral blood progenitors were induced to form myeloid colonies in the presence of conditioned medium by CSA-producing giant cell tumor (GCT) cells. Stimulation of NK cells with NK-insensitive targets failed to induce CIA production. Neutralizing antitumor necrosis factor (TNF) monoclonal antibodies (MoAbs) were found capable of inhibiting CIA present in the supernatants of NK cells stimulated with K562 cells. Following treatment with anti-TNF antibodies, CSA was again detectable in the same supernatants. This finding indicates that induction of TNF production did not concomitantly switch off CSA production by NK cells. Pretreatment of NK cells with recombinant interleukin-2 (rIL-2) or gamma interferon (r gamma IFN) did not change the amount of CSA released. However, treatment with rIL-2 caused the appearance of a factor in the NK cell supernatants capable of sustaining the formation of colonies of a larger size.

Production of Colony-Stimulating Activity by Human Natural Killer Cells: Analysis of the Conditions That Influence the Release and Detection of Colony-Stimulating Activity

AbstractHighly purified natural killer (NK) cell suspensions were tested for their capacity to release colony-stimulating activity (CSA) in vitro. NK cell suspensions comprised primarily CD16+ cells and were devoid of CD3+ T cells, CD15+ monocytes, and of B cells. CSA was detected in the NK cell supernatants and sustained the growth of myeloid colonies from both normal peripheral blood and bone marrow. CSA could be in part inhibited by pretreating NK cell culture supernatants with a specific goat anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) antiserum. The inhibition, however, was never complete, a finding that suggests that additional factors were responsible for CSA. Incubation of NK cells with K562 cells (an NK-sensitive target) or with normal bone marrow cells resulted in the appearance of a strong colony-inhibiting activity (CIA) in the culture supernatants. Such CIA was demonstrable in an experimental system where bone marrow or peripheral blood progenitors were induced to form myeloid colonies in the presence of conditioned medium by CSA-producing giant cell tumor (GCT) cells. Stimulation of NK cells with NK-insensitive targets failed to induce CIA production. Neutralizing antitumor necrosis factor (TNF) monoclonal antibodies (MoAbs) were found capable of inhibiting CIA present in the supernatants of NK cells stimulated with K562 cells. Following treatment with anti- TNF antibodies, CSA was again detectable in the same supernatants. This finding indicates that induction of TNF production did not concomitantly switch off CSA production by NK cells. Pretreatment of NK cells with recombinant interleukin-2 (rlL-2) or γ interferon (rγlFN) did not change the amount of CSA released. However, treatment with rlL-2 caused the appearance of a factor in the NK cell supernatants capable of sustaining the formation of colonies of a larger size.

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

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

Autocrine Growth Mechanisms of the Progenitors of Blast Cells in Acute Myeloblastic Leukemia

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

Hematopoietic growth factor production by cultured cells of human nasal polyp epithelial scrapings: Kinetics, cell source, and relationship to clinical status

The conditions and cell sources for colony stimulating activity (CSA) production by nasal polyp epithelial scrapings were examined. Epithelial scrapings removed from patients were grown to confluence during 7 days as monolayers of epithelial cells in media supplemented with fetal calf serum (FCS) on collagen-coated microwell plates. Growth kinetics of nasal polyp epithelial cells (NPECs) were determined, and CSA in NPEC conditioned medium (CM) was assessed with density-gradient separated, nonadherent peripheral blood mononuclear cells in standard 14-day methylcellulose assays. Nasal polyp cultures in the presence of 5% or 15% FCS (vol/vol) demonstrated significantly more epithelial cell proliferation than cultures at 0% and 1% FCS. There were comparable metachromatic cell counts in polyp epithelial scrapings from allergic and nonallergic donors. Similarly, NPEC CM from allergic and nonallergic donors had equivalent CSA for basophill mast cell (BMC) and eosinophil (EO) lineages, respectively. CSA production was enhanced under conditions of higher FCS concentration and NPEC proliferation. These studies confirm an epithelial cell origin of BMC and EO growth and differentiation factors derived from nasal polyps and point to the existence of a unique microenvironment for BMC and EO development provided by polyp epithelium that appears to be independent of the presence of an allergic diathesis.

Bone Marrow Stromal Cells in Myeloproliferative Disorders

The number of bone marrow-derived fibroblastoid colony-forming cells (CFU-F) and the production of colony-stimulating activity (CSA) by bone marrow stromal cells were studied in 71 patients with myeloproliferative disorders (MPD). The numbers of CFU-F in chronic-phase chronic myelogenous leukemia (CML), polycythemia vera (PV) and essential thrombocythemia (ET) were not different from those in normal subjects. However, the number of CFU-F in acute-phase CML was markedly decreased. Bone marrow adipocyte colony-forming capacity (adipo-CFC), which was previously shown to reflect both the number of preadipocytes and the stromal cell function in vivo, was increased in patients with chronic-phase CML, PV and ET, but was absent in acute-phase CML patients. The production of CSA by marrow stromal cells of MPD patients, however, was not different from that of normal subjects. These results suggest that the characteristics of marrow stromal and its precursor cells of chronic-phase MPD patients were not different from those of normal subjects, however, they became changed in acute-phase CML patients.

Factors influencing release of granulocyte-macrophage colony-stimulating activity from human mononuclear phagocytes.

Mononuclear phagocytes play an important role in the regulation of hematopoiesis, not only by producing regulatory monokines such as prostaglandins, tumor necrosis factor and interleukin-1 (IL-1), but also by the production of colony-stimulating activity (CSA). Previously, we have demonstrated that granulocyte-macrophage CSA (GM-CSA) production by mononuclear phagocytes can be induced by IL-1. In the present study, the influence of culture conditions on the production of GM-CSA was studied. It was found that both human sera and fetal bovine sera contain constituents - at present undefined - that induce GM-CSA production. These factors are distinct from IL-1 and lipopolysaccharide. In selected experiments, no GM-CSA-inducing effect of serum was found, suggesting that the effect may be donor-related. GM-CSA release in the presence of serum could be reduced by 40% after incubation of mononuclear phagocytes at low cell concentrations in methylcellulose, indicating that intimate cell-cell contact is an additional factor that enhances GM-CSA release.