Dexamethasone enhances macrophage colony stimulating factor- and granulocyte macrophage colony stimulating factor-stimulated proliferation of bone marrow-derived macrophages

Glucocorticoids are effective repressors of the immune system. We have examined the effect of glucocorticoids on the proliferation of murine macrophages. Dexamethasone by itself did not affect proliferation of differentiated or undifferentiated bone marrow-derived macrophages (BMM) and elicited peritoneal macrophages. However, dexamethasone enhanced the proliferation induced by macrophage colony stimulating factor (M-CSF) of these cells. The effect of dexamethasone was not restricted to M-CSF-dependent proliferation. Similarly, dexamethasone enhanced granulocyte macrophage colony stimulating factor (GM-CSF)- dependent proliferation of BMM. In agreement, macrophages transfected with the glucocorticoid receptor showed an enhancement of M-CSF- dependent proliferation. The enhancement of proliferation by dexamethasone or the glucocorticoid receptor was abolished by RU 486, an antagonist of the glucocorticoid receptor. Moreover, the addition of antibodies against M-CSF inhibits the effect of dexamethasone, suggesting that dexamethasone increases the autocrine production of M- CSF. This only occurs when M-CSF or GM-CSF, which induce M-CSF, are present in the media. In tissues, dexamethasone may enhance macrophage proliferation and contribute to the resolution of the inflammatory states.      

Different accessory function for TH1 cells of bone marrow derived macrophages cultured in granulocyte macrophage colony stimulating factor or macrophage colony stimulating factor.

The ability of macrophages to stimulate immune responses is heterogeneous and may have influence on the type of the developing immune response. Therefore, in an attempt to define different functional states of mouse macrophages, we made use of the two macrophage growth factors: macrophage colony stimulating factor (M-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF). Generation of macrophages from freshly isolated bone marrow cells in the presence of GM-CSF results in a population expressing profound antigen presenting function for mouse TH1 cells, resulting in strong lymphokine production and proliferation of the T cells. Furthermore, high amounts of a novel soluble cytokine active on mouse TH1 cells are generated during the interaction of TH1 cells with macrophages elicited with GM-CSF. In contrast, macrophages grown from bone marrow cells for at least 14 days in the presence of M-CSF express only minimal antigen-presenting function for TH1 cells. Treatment of such macrophages for 24 h with either IFN-gamma or GM-CSF allows the distinction between two further functional states. Those treated with IFN-gamma efficiently presented antigen towards TH1 cells. The T cells produced large amounts of lymphokines and proliferate well. However, synthesis of the novel soluble cytokine (active on TH1 cells) was not detectable. The generation of this mediator requires a short-term treatment with GM-CSF of macrophages developed in the presence of M-CSF prior to their interaction with TH1 cells.     

Comparison of bone marrow progenitors responsive to granulocyte-macrophage colony stimulating factor and macrophage colony stimulating factor-1

The responsiveness of bone marrow progenitors (BMP) from C3H mice to highly purified or recombinant preparations of Macrophage Colony-Stimulating Factor-1 (CSF-1) and Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) was compared by counting the number of colonies (greater than or equal to 50 cells) after 10 days in culture with CSF. Cells responsive to CSF-1 or GM-CSF exhibited maximum colony formation over a wide dose range, although GM-CSF supported colony formation at lower concentrations. The response of BMP to optimal concentrations of CSF-1 was greater than or equal to 5 times greater than the response of BMP to GM-CSF. Analysis of the kinetics of colony formation revealed that, at day 5, the number of BMP responsive to GM-CSF or CSF-1 was approximately equal; the number of CSF-1 colonies increased significantly through day 10, while those cultured in GM-CSF did not. The response of BMP to CSF-1 and GM-CSF was also studied in liquid culture; the differences in yield of mature macrophages was consistent with the differences observed in agar culture. Although both cell populations were shown to be 100% mononuclear by day 7, Coulter Channelyzer analysis of these mature macrophages showed marked differences in cell size distribution. By day 7, cells grown in CSF-1 resulted in a homogeneous population of large cells, whereas GM-CSF cultures showed a heterogeneous distribution. Finally, CSF-1-derived cells possessed increased nonspecific and specific phagocytic capabilities when compared to GM-CSF-derived macrophages. These findings indicate that the actions of GM-CSF and CSF-1 upon the bone marrow compartment results in the generation of mature macrophages which differ morphologically and functionally and may account for the heterogeneity in macrophage populations.     

Multiplexed particle-based anti-granulocyte macrophage colony stimulating factor assay used as pulmonary diagnostic test

Pulmonary alveolar proteinosis (PAP) is characterized by the accumulation of lipoproteinaceous material within the lung alveoli. Recent studies indicate that PAP is an autoimmune disease characterized by a neutralizing anti-granulocyte macrophage colony stimulating factor (GM-CSF) antibody. At present the only definitive diagnostic test for PAP is open lung biopsy. We have previously published that anti-GM-CSF is diagnostic for PAP and correlates with disease pathogenesis using a traditional serial anti-GM-CSF antibody titer format (T. L. Bonfield, M. S. Kavuru, and M. J. Thomassen, Clin. Immunol. 105:342-350, 2002). Titer analysis is a semiquantitative method, and often subtle changes in antibody titer are not detectable. In this report we present data to support anti-GM-CSF detection by a quantitative highly sensitive multiplexed particle-based assay which has the potential to be a clinical diagnostic test.     

重组人粒细胞巨噬细胞集落刺激因子对脓毒症模型小鼠的治疗作用

BACKGROUND: Granulocyte colony-stimulating factor has been shown to be used for the treatment of granulocytopenia, but its effect on sepsis is little reported. OBJECTIVE: To explore the treatment outcomes of recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) for sepsis mice. METHODS: A sepsis mouse model was established by intraperitoneal injection of lipopolysaccharide, and then given subcutaneous injection of rhGM-CSF at 6 and 30 hours after modeling. The morphological change of mouse lung tissue, CD64 expression in peripheral neutrophils, and serum levels of tumor necrosis factor-α and interleukin-10 were ovserved, respectively. RESULTS AND CONCLUISON:(1) Complete bronchial epithelial, mild stromal hyperplasia, and a few neutrophil infiltration were found after rhGM-CSF treatment. (2) CD64 expression in peripheral neutrophils, and serum level of interleukin-10 in the treatment group were significantly higher than those in the model and control groups at 1, 3 and 7 days after treatment (P < 0.05). The serum level of tumor necrosis factor-α in the treatment group was significantly higher than that in the control group, but lower than that in the model group after 1, 3 and 7 days of treatment (P < 0.05). (4) These results show that rhGM-CSF can enhance neutrophil function and the anti-inflammatory effects in sepsis mice. 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

Neurons and neuroblastoma as a source of macrophage colony-stimulating factor.

Accumulation of macrophages in brain tissue as observed in nervous system injury may be due to local production of hematopoietic colony-stimulating factors (CSF). The present work shows human neuroblastoma cells and murine neurons, namely granule cells of the cerebellum, to produce macrophage (M)-CSF which guides expansion and differentiation of macrophage lineage cells. The mRNA-encoding M-CSF but not the respective protein is present in mouse brain including cerebellum. Neither granulocyte M-CSF nor IL-3 is produced by cerebellar neurons or neuroblastoma. By their production of M-CSF, neurons may regulate the macrophage response and lead to local expansion and enhanced function of macrophages in inflammatory diseases of the central nervous system.     

Chemotactic peptides but not macrophage colony stimulating factor effect the synthesis of inositol lipids in macrophages.

Normal bone marrow derived macrophages display a wide variety of biological responses to a number of distinct agonists, for example, Macrophage Colony Stimulating Factor (M-CSF) and chemotactic peptides (such as FMLP). FMLP stimulates reactive oxygen intermediate production in these cells, whilst M-CSF stimulates DNA synthesis. We have compared the effects of these two agents on the production of novel inositol lipids in macrophages. Evidence is presented that FMLP, but not M-CSF elevate the levels of a lipid putatively identified as phosphatidylinositol-3,4-bisphosphate. The implications of this observation on proposed role of novel inositol lipids in macrophage proliferation are discussed.     

Ultrastructural effects of granulocyte colony stimulating factor (G-CSF) and macrophage colony stimulating factor (M-CSF) on rat neutrophils and monocytes

Human granulocyte colony stimulating factor (G-CSF) and macrophage colony stimulating factor (M-CSF) were administered intravenously to rats, and their effects on neutrophils and monocytes were examined by electron microscopy. G-CSF increased the number of cytoplasmic granules in neutrophils. It also enhanced maturation of the nuclear shape in the neutrophils, while chromatin condensation and peroxidase distribution remained immature. M-CSF induced proliferation of monocytes in peripheral blood and bone marrow, but did not affect morphology or distribution of peroxidase reactivity. This study was presented at the 25th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Matsumoto, September 28–30, 1993.     

Donated organ as a source of cytomegalovirus in orthotopic liver transplantation.

The importance of the donated organ as a source of CMV was assessed in 120 patients following orthotopic liver transplant and the CMV infections that developed in these patients were graded by severity. Forty-four recipients were CMV antibody negative pre-transplant. Eighteen of these received organs from CMV antibody positive donors and 15 (83%) developed primary CMV infections, 13 (87%) of which were symptomatic. Twenty-six received organs from CMV antibody negative donors and only 2 (8%) became CMV positive post transplant (P less than 0.001). These data suggest that there would be a considerable advantage in matching CMV antibody negative recipients with negative donors. Forty-five percent of secondary infections were asymptomatic compared with 12% of primary infections, and only 11% became disseminated compared with 53% of primary infections. The secondary infections that followed transplantation of an organ from a CMV antibody positive donor were more likely to be symptomatic and were more severe than those in patients who received seronegative livers.     

Granulocyte/macrophage colony stimulating factor. A potent activation signal for mature macrophages and monocytes

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a well-characterized hematopoietic growth factor. Recently, using purified recombinant-derived material, we have found that GM-CSF is also a potent activator of mature functional macrophages. Thus, we have found that exogenous GM-CSF augments the primary plaque-forming response to sheep red blood cells and that this effect is due to upregulation of Ia antigen expression and interleukin 1 production by the macrophages. We also show that GM-CSF inhibits the replication of Trypanosoma cruzi in cultured peritoneal macrophages and causes an accelerated clearance of Salmonella typhimurium from the peritoneal cavity of mice. These data indicate that GM-CSF is a multifunctional molecule stimulating both hematopoiesis and mature macrophage function.     

High colony stimulating activity in a patient with disseminated carcinoma and neutrophil leucocytosis

The case of a man with widespread bronchogenic carcinoma associated with pronounced neutrophil leucocytosis is presented. There was no evidence of infection or metastatic bone marrow infiltration. Increased levels of colony stimulating activity were shown in the patient's serum using three methods. Findings in this patient suggest that the leukaemoid blood picture was related to inappropriate tumour associated production of colony stimulating factors.     

Low serum conditions for in vitro generation of human macrophages with macrophage colony stimulating factor

Animal serum is often used to generate human macrophages in vitro. Since fetal calf serum (FCS) may complicate antigen uptake, processing and presentation on HLA molecules, we tested the ability of M-CSF to generate macrophages at low fetal calf serum conditions. Peripheral blood monocytes from 12 individuals were cultured 1-4 days with 0-100 ng/ml macrophage colony stimulating factor (M-CSF) at either 1 (low) or 5% (v/v) FCS. Regardless of number of days in culture, maximal (50-100 ng/ml) M-CSF stimulation and low FCS induced 65+/-5% esterase positive cells in all individuals compared to 52+/-7% without M-CSF (P<0.001). M-CSF increased the mean proportion of esterase positive cells after 24 or 96 h by 13% (P<0.005) and 13% (P<0.005), respectively, in 1% FCS, and 8% (P<0.05) and 2% (NS), respectively, in 5% FCS, indicating a slight negative interaction between 5% FCS and M-CSF (P<0.05). All cells were positive for CD14 and HLA class II, but cell number did not increase, confirming that M-CSF promote macrophage differentiation also at low FCS. M-CSF increased the average cell size after 24 or 96 h by 5.9+/-1.0 (P<0.05) and 8.6+/-0.5 (P<0.001) microm, respectively, without an increase in 5% FCS, further demonstrating the efficiency of M-CSF to promote macrophage generation at low FCS. The culture supernatants were negative for IL-1beta and TNF-alpha, which demonstrates that M-CSF did not activate the macrophages. The generation of human macrophages by M-CSF at low FCS should prove useful in studies where higher FCS concentrations may interfere with the assay.     

Presence and production of migration inhibitory activity in the peritoneal cavity.

The presence of migration inhibitory (MI) activity was investigated in the delayed hypersensitivity (DH) reaction induced by i.p. injection of PPD into FCA-sensitized guinea-pigs. Peritoneal exudates were studied at several times before and after the induction of the DH reaction and a non-immune control inflammation. Lymphokine (LK) fractions were prepared from individual exudates and tested in a conventional macrophage migration inhibition assay at concentrations determined to occur in vivo. The results suggest a continuous local production of MI activity and an augmented production during development of a DH reaction.     

骨调素和巨噬细胞集落刺激因子在糖尿病大鼠肾组织中的表达及霉酚酸酯的干预作用

目的： 研究骨调素(OPN)和巨噬细胞集落刺激因子（M-CSF)在糖尿病大鼠肾组织中的表达及免疫抑制剂霉酚酸酯（MMF）的干预作用，旨在探讨MMF对糖尿病肾病(DN)的保护作用及机制。方法: Wistar大鼠行右肾切除术2周后,随机分为右肾切除对照组(NC)、糖尿病组(DM)、霉酚酸酯治疗组(DM+MMF)。腹腔注射链脲佐菌素(STZ,65 mg/kg ) 诱发糖尿病模型,MMF15 mg·kg^-1·d^-1灌胃。检测各组8周末的左肾重/体重比值、24 h尿蛋白（Upro）、血糖(BG) 、血肌酐( Scr)，观察肾脏形态学变化，免疫组化检测肾组织中OPN、M-CSF及CD68表达，荧光实时定量PCR测定肾组织中OPN mRNA表达。结果: 与对照组相比，DM组大鼠血糖、Upro、肾重/体重比值均显著上升(P<0.01);肾间质纤维化面积扩大(P<0.01);肾组织内OPN、M-CSF、CD68表达及OPN mRNA的表达均显著上调(P<0.01)。MMF干预后，上述指标除血糖外均被明显抑制(P<0.05或P<0.01)。结论: MMF减少糖尿病大鼠肾组织中OPN、M-CSF、CD68及OPN mRNA的表达，降低蛋白尿，预防肾损伤。MMF明显抑制DN肾小管－间质损害，可能与其抑制巨噬细胞的趋化与增殖有关。     

异型巨噬细胞集落刺激因子在人白血病细胞系中的表达及性质

目的 探讨异型巨噬细胞集落刺激因子（Ｍ－ＣＳＦ）在人白血病细胞系中的表达和性质。方法 采用ＡＢＣ免疫酶标技术,间接免疫荧光染色,流式细胞计数,蛋白免疫印迹和反向酶联ＤＮＡ蛋白质相互作用分析技术研究了４株人白血病细胞系（Ｊ６－１、Ｊ６－２、Ｋ５６２、ＨＬ６０）和正常人外周血单个核细胞Ｍ－ＣＳＦ的分布、表达及其分子大小。结果 正常人外笛血单个核细胞示见Ｍ－ＣＳＦ的表达,经植物血浆素刺激后有低水平的表达