Effect Of M-CSF Research Articles

Effect of M-CSF on the expression of endothelial progenitor cell markers in blood mononuclear cell culture in coronary heart disease

Aim. To evaluate the nature of changes in the expression of markers of endothelial progenitor cells (VEGFR2, CD34, CD14) and endothelial cells (CD146) in association with the expression of the leukocyte common antigen CD45 in the culture of blood mononuclear cells in the presence of M-CSF in patients with coronary heart disease (CHD) and healthy donors.Materials and methods. The study included 12 patients with CHD with class III–V angina pectoris and 10 healthy donors, from whom 30 ml of venous blood was taken on an empty stomach in the morning and stabilized with heparin. Blood mononuclear cells were isolated by Ficoll density gradient centrifugation (1.077 g / cm3) and subject to immunomagnetic separation using CD14-MicroBeads and CD34-MicroBead Kit (Miltenyi Biotec B.V. & Co. KG, Germany). The resulting CD14+ and CD34+ culture of mononuclear cells was incubated for 6 days in a complete nutrient medium with and without M-CSF 50 ng / ml (Cloud-Clone Corp., USA) with complete replacement of the medium and repeated application of M-CSF on day 3. After 6 days, the proportions of CD45+, CD14+, CD34+, VEGFR2+, and CD146+ cells in the culture were assessed by flow cytometry using CD14-FITC, CD34-PE, VEGFR2-Alexa Fluor 647; CD45-FITC and CD146-PerCP antibodies (BD Biosciences, USA).Results. It was shown that in healthy donors, the proportion of CD146+ cells in the co-culture of blood mononuclear cells with M-CSF exceeded their number in the sample without it, with comparable expression rates of CD45, CD14, and VEGFR2 markers between the control and stimulated cultures. In CHD patients, the number of CD146+ and VEGFR2+ cells did not change when M-CSF was added to the mononuclear cell culture; however, the proportion of CD14+ cells increased and the proportion of CD45+ cells decreased compared to the control sample. The number of CD34+ cells was comparable both between control and stimulated samples, and between the groups of examined individuals. At the same time, in patients with CHD, an increased proportion of VEGFR2+ cells was found in the control and stimulated samples compared to healthy individuals, while an increased proportion of CD14+ cells was detected only in the stimulated culture.Conclusion. The development of CHD disrupts the response of blood mononuclear cells to the effect of M-CSF, increasing the number of CD14+ and reducing the proportion of CD45+ cells in the culture in the absence of stimulating effects on the expression of endothelial cell marker CD146. At the same time, M-CSF does not affect the expression of CD34 and VEGFR2 in endothelial progenitor cells both in patients with CHD and in healthy individuals.

ASBMR 29th Annual Meeting

The critical role of M-CSF in the differentiation of osteoclasts (OC) is well established. In mature OC, M-CSF has been reported to regulate survival and motility, but no clear role in resorptive function has been established. In this study we investigated the role of M-CSF on resorbing activity of mature human OC. Human OC were generated by treatment of CFU-GM-derived precursors with soluble hRANKL (125ng/mL) and hM-CSF (25ng/mL) for 14–21d. To quantify resorbing activity, mature OC were dissociated from the plastic substrate using a non-enzymatic buffer, settled onto dentine slices and cultured for 72h. Activation of NFκB and AP-1 was assessed using a sandwich ELISA technique. IκBα and ERK were assessed by Western analysis. Neither M-CSF nor RANKL were required for mature OC survival over 72h. However, resorption function was absolutely dependent on the addition of RANKL. Co-treatment with M-CSF had a concentration-dependent biphasic action, greatly augmenting RANKL-induced resorbing activity (+181%) and actin ring formation (+131%) at 25ng/mL. RANKL alone activated NFκB (+554%). M-CSF had no effect on NFκB but potentiated RANKL-induced activation (+42%). In contrast, RANKL alone had no effect on AP-1 activation whereas M-CSF stimulated weakly (+63%) and the combination produced a synergistic effect (+171%). M-CSF activated phosphorylation of ERK 1/2, peaking at 5 min, whereas RANKL had no effect alone or in combination with M-CSF. M-CSF had no effect on IκBα phosphorylation whereas RANKL alone stimulated, with no additional effect in combination with M-CSF. Time-lapse video-microscopy showed no apparent effect of M-CSF on the motility of OC co-treated with RANKL. We have demonstrated that M-CSF is not required for survival of mature human OC, but it does potently modulate RANKL-induced resorbing activity via NFκB and AP-1 activation. The molecular mechanism of this effect remains to be determined although we propose a convergence of RANK and fms signaling (“cross-talk”) upstream of the critical OC transcription factors NFκB and AP-1. These findings have important implications for the clinical management of diseases associated with excess bone resorption.

Effect of M-CSF, GM-CSF in adenovirus-mediated gene transfer into macrophages

Effect of M-CSF, GM-CSF in adenovirus-mediated gene transfer into macrophages

Does macrophage-colony stimulating factor stimulate rat haematopoietic cells?

It is known that colony stimulating factors (CSFs) stimulate the myeloid cells of bone marrow and splenic cells in rodents. The effects of macrophage (M)-CSF on the activities of thymidylate synthase and thymidine kinase, involved in de novo and salvage pathways for pyrimidine nucleotide synthesis, respectively, in haematopoietic cells of bone marrow and spleen were investigated in rats. A single M-CSF injection did not elevate the mRNA expression levels of the enzymes in bone marrow cells 6 h after treatment, but it enhanced the splenic thymidylate synthase mRNA expression. M-CSF stimulated the splenic thymidylate synthase activity without an increase of the peripheral granulocytes. The effect of M-CSF on granulocytes is considered to be weak compared with that of granulocyte (G)-CSF, because of the indirect secretion of endogenous G-CSF from the cells with M-CSF receptors stimulated by exogenous M-CSF. Since M-CSF was able temporarily to lead progenitor cells from long G1-phase into S-phase, M-CSF might accelerate the anticancer effects when used together with anticancer agents.

Changes in M-CSF-like activity during chicken embryonic development

We have established a method for separation of chicken bone-marrow cells using Percoll density gradient centrifugation, and have developed a new method for determining chicken M-CSF-like activity employing a liquid culture. Using this method, we determined M-CSF-like activities in egg yolk, chorioallantoic fluid (CAF) and amniotic fluid (AmF), and studied the effects of M-CSF on development of chicken embryos. M-CSF-like activity in egg yolk was at a high level before the incubation of the egg; it began to decrease on the third day of incubation and rapidly decreased on the fourth day, and no significant activity was detected after the tenth day of incubation. M-CSF-like activity in CAF was very low, and it exhibited almost no change during development. No M-CSF-like activity was detected in AmF throughout the experimental period.

Cytokine and growth factor regulation of macrophage scavenger receptor expression and function

Regulation of macrophage scavenger receptor (MSR) activity may be an important determinant of the extent of atherogenesis and the efficacy of host defense. The effect of M-CSF on this pathway was studied using a recently developed monoclonal antibody to murine MSR. M-CSF markedly and selectively increased MSR synthesis in murine macrophages (Mφ); post-translationally the receptor appeared more stable and shifted to a predominantly surface distribution. Functionally M-CSF enhanced modified lipoprotein uptake and increased divalent cation-independent adhesion in vitro. These results suggest a plausible mechanism whereby M-CSF production in the atheromatous plaque micro-environment could promote the recruitment and retention of mononuclear phagocytes and subsequent foam cell formation. In addition, the Th1 cytokine (γ-interferon) and Th2 cytokine (interleukin-4) had differential effects on MSR glycosylation in vitro suggesting a further possible regulatory role by these lymphokines on macrophage MSR function.

Antitumor effects of human recombinant macrophage colony-stimulating factor against rat brain tumors.

The tumoricidal effects of M-CSF were examined using two subcutaneously-transplanted rat brain tumor cell lines, 9L and T9 gliomas. In rats treated with high-dose M-CSF (16 million U/kg administered for 4 days a week for 3 weeks), 9L glioma growth was inhibited by 81.9% following subcutaneous (s.c.) injection and by 70.5% after intraperitoneal (i.p.) injection and T9 glioma growth was inhibited by 69.2% after i.p. injection. After short-term treatment with high-dose M-CSF (32 million U/kg administered s.c. for 6 consecutive days, 9L glioma growth was inhibited by 82.1%. All these inhibitory effects differed significantly compared with the respective untreated control groups. However, treatment with low-dose M-CSF (1.6 million U/kg administered s.c. for 4 days a week for 3 weeks) showed no significant effects against 9L and T9 glioma growth compared with the untreated controls. No significant effects of M-CSF against cell proliferation, measured as PCNA expression, were observed in any group. Significant hematopoietic effects on the leukocyte counts were observed only in the groups treated with high dose M-CSF. These results suggest that M-CSF at a high dose which produces hematopoietic effects on peripheral leukocytes inhibits the growth of gliomas. This inhibitory effect may have been due to a tumoricidal mechanism of M-CSF that depended on the production or release of some hematopoietic soluble factors, but was independent of PCNA expression by the tumors.

Macrophage Colony-Stimulating Factor and Granulocyte-Macrophage Colony-Stimulating Factor Stimulate the Synthesis of Plasminogen-Activator Inhibitors by Human Monocytes

AbstractMacrophage colony-stimulating factor (M-CSF or CSF-1) and granulocyte-macrophage CSF (GM-CSF) have been shown to increase human monocyte urokinase-type plasminogen-activator (u-PA) activity with possible consequences for cell migration and tissue remodeling; because monocyte u-PA activity is likely to be controlled in part also by the PA inhibitors (PAIs) made by the cell, the effect of M-CSF and GM-CSF on human monocyte PAI-2 and PAI-1 synthesis was investigated. To this end, elutriation-purified human monocytes were treated in vitro with purified recombinant human M-CSF and GM-CSF, and PAI-2 and PAI-1 antigen and mRNA levels measured by specific enzyme-linked immunosorbent assays and Northern blot, respectively. Each CSF could enhance the protein and mRNA levels of PAI-2 and PAI-1 at similar concentrations for each product. This similar regulation of monocyte PAI expression in response to the CSFs contrasted with that found for the effects of lipopolysaccharide, transforming growth factor-β and a glucocorticoid. Therefore, PAIs may be modulating the effects of the CSFs on monocyte u-PA activity at sites of inflammation and tissue remodeling.

Macrophage colony-stimulating factor and granulocyte-macrophage colony- stimulating factor stimulate the synthesis of plasminogen-activator inhibitors by human monocytes

Macrophage colony-stimulating factor (M-CSF or CSF-1) and granulocyte- macrophage CSF (GM-CSF) have been shown to increase human monocyte urokinase-type plasminogen-activator (u-PA) activity with possible consequences for cell migration and tissue remodeling; because monocyte u-PA activity is likely to be controlled in part also by the PA inhibitors (PAIs) made by the cell, the effect of M-CSF and GM-CSF on human monocyte PAI-2 and PAI-1 synthesis was investigated. To this end, elutriation-purified human monocytes were treated in vitro with purified recombinant human M-CSF and GM-CSF, and PAI-2 and PAI-1 antigen and mRNA levels measured by specific enzyme-linked immunosorbent assays and Northern blot, respectively. Each CSF could enhance the protein and mRNA levels of PAI-2 and PAI-1 at similar concentrations for each product. This similar regulation of monocyte PAI expression in response to the CSFs contrasted with that found for the effects of lipopolysaccharide, transforming growth factor-beta and a glucocorticoid. Therefore, PAIs may be modulating the effects of the CSFs on monocyte u-PA activity at sites of inflammation and tissue remodeling.