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Results for “Granulocyte Precursor Cells”

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PubMed1992

Regulation of L-selectin expression on cultured bone marrow leukocytes and their precursors.

L-selectin (LECAM-1, LAM-1, MEL-14 antigen, Dreg antigen) is one of the molecules controlling lymphocyte homing from the blood to peripheral lymph nodes and granulocyte adhesion to inflamed endothelium. In this work, regulation of L-selectin expression on mouse bone marrow cells was studied. L-selectin-negative cells were isolated by panning technique, cultured for 1-7 days with cytokines and mitogens, and L-selectin expression was analyzed by immunofluorescence staining. When cultured for 3 days with interleukin (IL) 1, IL 2, IL 5, IL 6, phytohemagglutinin, pokeweed mitogen or in the medium alone, 75%-85% of L-selectin-negative large cells (including granulocytes, macrophages/monocytes, blasts and their precursors) became L-selectin positive. In contrast, IL 3, IL 4, granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) prevented the induction of L-selectin in a time- and dose-dependent manner. GM-CSF was the most potent inhibitor and only 10%-15% of cells became L-selectin positive after 3 days of culture. Furthermore, L-selectin was down-regulated on cultured unselected bone marrow cells by IL 3, IL 4, GM-CSF and LPS stimulation. After culture, the relative molecular mass of L-selectin was 100 kDa, similar to the size of the granulocyte form of this antigen. Cultured cells adhered to high endothelial venules (HEV) only 10%-32% as effectively as freshly isolated bone marrow cells despite high levels of L-selectin expression. The phenotypic analysis and the HEV binding data indicate that after culturing L-selectin was almost exclusively expressed on bone marrow leukocytes of myeloid series, and on these cells it was not functional in mediating peripheral lymph node HEV binding. Overall, these results show that the expression of L-selectin can be modulated by regulating the maturation and differentiation of the cells in vitro. This supports the idea that different cytokines and mitogens may also be important in controlling migrational status of leukocytes in vivo.

Animals↗

PubMed2004

Local mechanisms underlying the regulatory effect of Kropanol on hemopoiesis during paradoxical sleep deprivation.

We studied the effect of Kropanol on local regulatory mechanisms of hemopoiesis during paradoxical sleep deprivation. The regulatory effect of Kropanol on granulomonocytopoiesis was determined by increased binding capacity of stromal cells in relation to granulocyte-macrophage precursors and increased formation of mixed hemopoietic islets leading to accelerated maturation of granulocyte-macrophage precursors and hyperplasia of the granulomonocytic hemopoietic stem. Stimulation of erythropoiesis with Kropanol was associated with increased formation of erythroid hemopoietic islets, accelerated differentiation of erythroid precursors, and high content of erythroid cells in the bone marrow. Kropanol increased proliferative activity of erythroid precursors. Stimulation of these processes depended on enhanced production of short-distance humoral regulators of erythro- and granulomonocytopoiesis.

Animals↗

PubMed1992

Stem cell factor stimulates the in vitro growth of bone marrow cells from aplastic anemia patients.

Aplastic anemia (AA) is a rare human bone marrow disorder of unknown etiology manifested by a strongly impaired growth of hematopoietic precursors. In this study, we examined the ability of recombinant human stem cell factor (SCF) to stimulate proliferation in vitro of bone marrow cells from 15 AA patients. All patients had been previously treated with antilymphocyte globulin (ALG). SCF, in combination with erythropoietin (Epo), interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF), increased the number of hematopoietic colonies formed in a semisolid medium by AA marrows. Maximal colony numbers reached 30% of the numbers observed with normal bone marrow cells. Proliferation of AA cells cultured in a liquid medium containing SCF together with Epo, IL-3, GM-CSF, and G-CSF approached 70% of the control level, as measured by 3H-thymidine incorporation. The effect of the combination of SCF with the other growth factors was more than 10 times stronger than that of the growth factors alone. The most marked effect of SCF was on the generation of erythroid colonies by precursor cells. The results demonstrate synergism between CSF and other hematopoietic growth factors, resulting in the most efficient stimulation of the in vitro growth of AA bone marrow cells described to date. Use of SCF, either alone or in combination with other factors, may be of potential value in treatment of AA.

Adolescent↗

PubMed1982

Culture of macrophage cell lines from normal mouse bone marrow.

The use of semisolid medium for the culture and cloning of haematopoietic cells has helped our understanding of their proliferation and differentiation. It has been shown that mixed colonies of granulocytes and macrophages developed, in the presence of colony-stimulating factor (CSF), from their common precursor, granulocyte-macrophage colony-forming cells (GM-CFC). Attempts at recloning these colonies in semisolid medium suggested that granulocytes and macrophages were differentiated cells incapable of further proliferation. However, our studies on cultures of larger numbers of cells demonstrate that while this may be the case for granulocytes, macrophages seem to be capable of long-term proliferation.

Animals↗

PubMed2001

Generation of dendritic cells from leukaemia cells of a patient with acute promyelocytic leukaemia by culture with GM-CSF, IL-4 and TNF-alpha.

Dendritic cells are potent antigen-presenting cells derived from CD34+ haemopoietic stem cells. Dendritic cells have been reported to be generated from cells in granulocytic lineage as well as monocytes, blood dendritic cell precursors and lymphoid progenitors. In order to explore the differentiation pathway of dendritic cells from granulocytic cells and the applicability of leukaemia-derived dendritic cells for anti-leukaemic immunotherapy in acute leukaemia of granulocytic origin, we tried to generate dendritic cells from leukaemia cells of a patient with acute promyelocytic leukaemia (APL). Leukaemia cells were cultured with GM-CSF, IL-4 and TNF-alpha for 10 days. Azurophilic granule-containing cells with marked cytoplasmic projections were generated in the culture. FACS analysis of these cultured cells revealed the generation of CD1a+, CD83+, CD80+, CD86+, CD40+ and HLA-DR+ cells. The leukaemic origin of these dendritic-like cells was demonstrated by in situ hybridization of magnetic-bead-sorted CD1a+ dendritic cells using the DNA probes of t(15;17). Cells generated by culturing leukaemia cells were demonstrated to have a potent antigen-presenting function in allogeneic mixed leucocyte cultures. These findings show the plausibility of the previously reported pathway of dendritic cell maturation through granulocytic cells and suggest the possibility of anti-leukaemic immunotherapy using leukaemia-derived dendritic cells even in patients with acute promyelocytic leukaemia.

Adult↗

PubMed2004

[Granulocyte-colony stimulating factor (G-CSF) in the plasma and activity of enzymes in granulocytes of breast cancer patients].

OBJECTIVES: G-CSF is a stimulatory factor of granulocyte colony formation. In vitro and in vivo G-CSF stimulates the proliferation of granulocyte precursors and enhances phagocytic functions of mature cells. Indirect evidence of this might be the increase of the activity of granulocyte enzymes participating in phagocytosis. DESIGN: The aim of this study was to compare in vivo the level of G-CSF in the plasma and the activity of: alkaline phosphatase (AP), acid phosphatase(AcP), peroxidase(MPO) and esterase in the granulocytes of breast cancer patients. MATERIALS AND METHODS: The activities of tested enzymes were measured by cytochemical methods in the blood of 10 patients with breast cancer before and 30 days after operation and in 10 healthy subject. GSF level was measured using a sensitive sandwich ELISA. CONCLUSIONS: 1. On the basis of the obtained results we can conclude that G-CSF level was increased in cancer patients (18.30 pg/ml before operation and 20.70 pg/ml after operation) compared to the control group (15.73 pg/ml). This results suggest that G-CSF may be synthesized by phagocytic cells and/or cancer cells. 2. AP activity was decreased after surgery (135.00 score) in comparison to the activities before operation (182.3 score). 3. AcP (213.6 score), MPO (364.6 score) and esterase (184.2 score) activities were significantly increased after surgery in comparison to the activities before operation: AcP 188.6 score, MPO 316.3 score, esterase 154.8 score. 4. These results suggest that activities of tested enzymes might be dependent on G-CSF plasma level.

Acid Phosphatase↗

PubMed1993

Microenvironmentally dependent effects on murine haemopoiesis by a prolonged interleukin-1 treatment.

We administered recombinant human IL-1 beta (400 ng/d, s.c.) for 10 d to normal C57B1 mice and determined daily granuloid and erythroid parameters in marrow, spleen and blood. In the marrow CFU-GM numbers were not affected but later granuloid cell stages were moderately enhanced (170%). In the spleen, however, CFU-GM numbers were sharply increased (1600%), whereas the granuloid precursors only doubled. Blood granulocytes increased transiently to 275% on day 5. In the marrow all erythroid parameters were severely reduced. This reduction was partially compensated by the spleen where initially only BFU-E and with some delay also more mature erythroid cells accumulated. At the end of the treatment mice were slightly anaemic. When mice were treated with IL-1 and erythropoietin (10 U/d) simultaneously, the inhibitory effects on erythropoiesis were less severe. In agreement with in vivo results, IL-1 inhibited in vitro colony growth of CFU-E from normal bone marrow and spleen but spleen CFU-E from 5 d IL-1 treated mice were insensitive. We conclude that IL-1 can induce stimulation or inhibition of haemopoietic progenitor cells depending on their microenvironment.

Animals↗

PubMedDate not supplied

Immunopathogenesis of chicken anemia virus infection.

The immunopathogenesis of chicken anemia virus (CAV) infection is reviewed. The virus causes a disease in young chicks which is characterised by generalised lymphoid atrophy, increased mortality and severe anemia. The virus appears to target erythroid and lymphoid progenitor cells in the bone marrow and thymus respectively. The B cells in the chicken are not susceptible to CAV infection and are not directly affected by the virus. Destruction of erythroid progenitors in bone marrow results in severe anemia, and depletion of granulocytes and thrombocytes. Destruction of precursor T cells results in depletion of mature cytotoxic and helper T cells with consequent effects on susceptibility to, and enhancement of, the pathogenicity of secondary infectious agents, and sub-optimal antibody responses. Apoptosis appears to be a feature of the lymphocyte depletion in the thymic cortex, which may be mediated by one of the non-structural viral proteins, VP3 (apoptin).

Animals↗

PubMed1999

Differential activity of glycosaminoglycans on colony-forming cells from cord blood. Preliminary results.

Heparin, heparan sulfate and chondroitin sulfate were evaluated for their possible role on proliferation and differentiation of hematological precursor cells from cord blood. For these purposes, different concentrations of glycosaminoglycans were added to methyl-cellulose in colony assay performed with human cord blood derived cells. A volume of 10 microg/ml heparin induces a significant increase of both granulocyte-monocyte and granulocyte colonies, and a decrease of erythroid-colonies, more evident in the presence of 100 microg/ml. Heparan sulfate-treatment induces a significant increase of all granulocyte-monocyte colonies derived from CFU-granulocyte-monocyte, CFU-granulocyte and CFU-monocyte precursors. A significant decrease of multipotent cells was also observed. On the other hand, chondroitin sulfate induces an increase of granulocyte-colonies and a decrease of erythroid-colonies. Glycosaminoglycans with different structure may be useful to increase the number of specific colonies. The selective and differential binding of glycosaminoglycans with several growth factors and the regulation of their activities is discussed.

Fetal Blood↗

PubMed1994

Stem cell factor (c-kit ligand) enhances the interleukin-9-dependent proliferation of human CD34+ and CD34+CD33-DR- cells.

We have studied the effects of recombinant human interleukin-9 (IL-9), alone and combined with stem cell factor (SCF, c-kit ligand), IL-3, and granulocyte-macrophage colony-stimulating factor (GM-CSF) on the clonogenic proliferation of highly enriched human hematopoietic CD34+ and CD34+CD33-DR- progenitor cells. Colony assays were performed under serum-containing and serum-free conditions. IL-9, as a single agent, did not support colony formation. The addition of erythropoietin (Epo) to IL-9 induced the growth of erythroid progenitors (BFU-E) derived from both CD34+ and CD34+CD33-DR- cells. The IL-9-dependent growth of BFU-E derived from CD34+ cells was increased in an additive manner by SCF and, to a lesser extent, by IL-3, whereas CD34+CD33-DR- erythroid precursors were also responsive to GM-CSF in combination with IL-9. The addition of SCF to IL-9 did stimulate the development of CD34+ and CD34+CD33-DR- macroscopic, multicentered BFU-E and multilineage colonies (CFU-GEMM). When IL-9 was used in serum-free conditions, the growth of CD34+ and CD34+CD33-DR- BFU-E was observed in the presence of Epo. Moreover, a marked synergy on BFU-E colony formation was evident when IL-9 was combined with SCF, and their activity was enhanced by the addition of IL-3. IL-9 showed a negligible proliferative activity on colony-forming units-granulocyte/macrophage (CFU-GM). However, it increased the number of CD34+CD33-DR- CFU-GM responsive to IL-3 (37% of the colonies generated by phytohemagglutinin-stimulated lymphocyte conditioned medium [PHA-LCM]). The effects of IL-9 on CD34+CD33-DR- cells were also studied in a short-term suspension culture system, which evaluates the proliferation of progenitors earlier than day 14 CFU-C (Delta assay). In this system, IL-9 had a minimal activity on its own. In combination with SCF, however, it induced a nine-fold expansion of CD34+CD33-DR- cells, which generated a greater number of CFU-GM than BFU-E in secondary methylcellulose cultures. These experiments indicate that IL-9 induces the proliferation of very primitive human erythroid cells, and this effect is potentiated by SCF and other cytokines. Furthermore, IL-9 synergizes in vitro with the c-kit ligand in expanding the pool of early pluripotent hematopoietic progenitor cells.

Antigens, CD↗

PubMed1983

Characterization of mast cell precursors by physical means: dissociation from T cells and T cell precursors.

Murine bone marrow precursors (MCP) differentiate into mast cells and proliferate in response to mast cell growth factor (MCGF). An assay system based on the incorporation of [3']thymidine by proliferating mast cells in response to substantially purified MCGF was used to titrate MCP. Murine bone marrow was separated into fractions by Percoll density gradient centrifugation or velocity sedimentation at unit gravity. Individual fractions were analyzed to determine the relative concentrations of MCP. To determine if mast cells were derived from precursors different from those for T lymphocytes and granulocytes and macrophages, separated cells were also analyzed for responsiveness to interleukin 2 (IL 2) in standard [3H] thymidine incorporation assays and to granulocyte-macrophage colony stimulating factors (CSF) in agar cloning (CFU-GM) assays. Cultured marrow cells from Dexter type long-term marrow cultures (LMC) were included as a source of marrow cells devoid of mature T cells, so as to dissociate MCP from pre-T cells. MCP were readily dissociable from T cells and/or pre-T cells present in fresh, as well as cultured, marrow cells by either density or velocity of sedimentation separation techniques. MCP were, however, not readily separable from CFU-GM. Nevertheless, MCP appeared not to share a common precursor with pure macrophage type colony-forming cells (CFU-M phi). The implications of the findings on the lineage origin of mast cells were discussed.

Animals↗

PubMed1983

Presumed myelopoiesis in spinal dura mater of normal bovine foetuses.

In the course of a study of normal bovine foetal development, the brain and spinal cord of 24 newborn calves and 81 foetuses, all of the Jersey breed and ranging in age from 60 to 280 days gestation, were examined histologically. This examination revealed cellular aggregates in the spinal dura mater of all newborn calves and all foetuses over 119 days gestation. The cells had the appearance of blood leucocytes (granulocytes, monocytes) and their precursors; and also, in the foetuses only, giant cells identified as megakaryocytes were also seen. It is suggested that in the second half of gestation and for at least 2 days after birth, the spinal dura mater of cattle, in the Jersey breed at least, acts as a site of myelopoiesis. The importance of this finding to veterinary pathologists is emphasized.

Animals↗

PubMed2006

The Integrin alpha9beta1 contributes to granulopoiesis by enhancing granulocyte colony-stimulating factor receptor signaling.

The integrin alpha9beta1 is widely expressed on neutrophils, smooth muscle, hepatocytes, endothelia, and some epithelia. We now show that mice lacking this integrin have a dramatic defect in neutrophil development, with decreased numbers of granulocyte precursors in bone marrow and impaired differentiation of bone marrow cells into granulocytes. In response to granulocyte colony-stimulating factor (G-CSF), alpha9-deficient bone marrow cells or human bone marrow cells incubated with alpha9beta1-blocking antibody demonstrated decreased phosphorylation of signal transducer and activator of transcription 3 and extracellular signal-regulated protein kinase. These effects depended on the alpha9 subunit cytoplasmic domain, which was required for formation of a physical complex between alpha9beta1 and ligated G-CSF receptor. Integrin alpha9beta1 was required for granulopoiesis and played a permissive role in the G-CSF-signaling pathway, suggesting that this integrin could play an important role in disorders of granulocyte development and other conditions characterized by defective G-CSF signaling.

Animals↗

PubMed1979

Chronic myelomonocytic leukemia: light and electron microscopy of the bone marrow.

Clinical data and light and electron microscopic findings are presented in a patient with chromic myelomonocytic leukemia of about 5 years' duration and no need for specific therapy. Cytogenetic studies failed to demonstrate a Philadelphia-chromosome. The leading clinical symptoms were anemia, moderate hepatomegaly, and leukocytosis with monocytes in the peripheral blood count. Light microscopy of bone marrow cores showed hypercellularity of neutrophil granulocytic and monocytic cell lines including some precursor forms. Electron microscopy confirmed the existence of a biphasic myelomonocytic cell proliferation with predominance of mature forms in both lineages; there were no gross cellular abnormalities and no "hiatus leukaemicus". Consupicuous were cells of an undeterminated origin apparently neither belonging to the neutrophil granulocytic nor monocytic series and large histiocytic cells, possibly corresponding to the so-called sea-blue histiocytes of light microscopy. The high degree of maturation of both cell lines in the bone marrow is in accordance with the relatively benign and prolongated course of this rare type of leukemia.

Aged↗

PubMed1995

Progenitor cells in the blood and marrow of patients with chronic phase chronic myeloid leukaemia respond differently to macrophage inflammatory protein-1 alpha.

Macrophage inflammatory protein-1 alpha (MIP-1 alpha) is a negative regulator of normal haemopoietic stem cell proliferation. Insensitivity to MIP-1 alpha of progenitor cells in chronic myeloid leukaemia (CML) could, therefore, explain myeloid expansion in this disease. We compared the effects of MIP-1 alpha on progenitor cells in normal marrow and in the blood and marrow of patients with chronic phase CML. Plastic-adherent precursors of granulocyte-macrophage colony-forming cells (P delta progenitors) are very primitive progenitor cells and are detected by incubating them for 1 week in liquid culture and assaying the CFU-GM released into the supernatant. Direct CFU-GM assays were also used in this study. Daily addition of 300 ng/ml/day of MIP-1 alpha to P delta progenitor assays of normal marrow cells suppressed CFU-GM production by 50% and in CML bone marrow P delta cultures by 20-30%. The response of CML blood P delta progenitors was heterogeneous. In five of nine cases, CFU-GM production was doubled in the presence of MIP-1 alpha and in four of nine cases, it was reduced. Addition of 100-500 ng MIP-1 alpha to direct assays of CFU-GM stimulated colony formation by normal marrow and CML blood cells to a similar extent.

Bone Marrow Cells↗

PubMedDate not supplied

Misonidazole enhances cyclophosphamide toxicity to bone marrow.

Normal bone marrow function was surveyed in animals that received cyclophosphamide after treatment with small, multiple doses of misonidazole. Peripheral blood white cell counts, cell differentials, hematocrits, bone marrow white cell counts and committed granulocyte-macrophage precursors (CFUc) were monitored for several days following treatment with misonidazole and cyclophosphamide. When results obtained from animals treated with physiological saline or misonidazole in advance of cyclophosphamide were compared, no significant differences were noted in routine assays of white blood cell count, cell morphology or hematocrit. Pre-treatment with misonidazole, however, caused a significant reduction in survival and delay in recovery of bone marrow CFUc (p less than .01). The combined use of misonidazole and cyclophosphamide also reduced animal survival (DMF = 1.2), with the majority of deaths being attributable to failure of normal hematopoietic function. These results suggest that misonidazole enhances the myelotoxicity associated with cyclophosphamide use. This additional damage is not detectable using routine hematological assays, but is demonstrable in assays of bone marrow stem cells and animal survival studies.

Animals↗

PubMed1998

Leukemic cells with 11q23 translocations express granulocyte colony-stimulating factor (G-CSF) receptor and their proliferation is stimulated with G-CSF.

We report a 20-month-old boy with acute lymphoblastic leukemia with the 11q23 translocation whose blasts markedly increased in peripheral blood after intravenous granulocyte colony-stimulating factor (G-CSF) administration, but disappeared after stopping G-CSF. The in vitro study showed that the leukemic cells separated from this patient expressed G-CSF receptor (G-CSFR) and an addition of G-CSF stimulated their proliferation by 3H-thymidine incorporation assay (stimulation index, 4.9). To clarify whether or not leukemic cells with 11q23 translocations generally express G-CSFR and show proliferative response to G-CSF, we performed the similar in vitro experiments using eight leukemic cell lines with 11q23 translocations. We found that all cell lines examined expressed G-CSFR (20-98%) and proliferation of seven leukemic cell lines was significantly enhanced in response to G-CSF (stimulation index >1.5 in five cell lines), suggesting a possible participation of the G-CSF/G-CSFR interaction in the process of growth regulation of leukemic cells with 11q23 translocations.

Antineoplastic Combined Chemotherapy Protocols↗

PubMed1980

Effect of daily administration of cytotoxic drugs on the erythroid and granulocytic repopulating ability of rat bone marrow.

The erythroid and granulocytic repopulating ability of rat bone marrow has been measured after daily administration of three alkylating agents: dimethylmyleran, busulphan and cyclophosphamide. Three different patterns of response were observed. The results emphasize the variability of the response of haemopoietic tissue to repeated administration of different cytotoxic drugs and provide further evidence that erythroid and granulocytic repopulating assays measure different populations of precursor cells.

Animals↗