CSF-1 Production Research Articles

Endogenous CSF-1 Production is Associated with Viral Replication in HIV-1 Infected Human Monocytes

GRUBER, MARION F.; GERRARD, THERESA L.; FAUST, KARIS; BOONE, ELIZABETH; CLOUSE, KATHLEEN A. Author Information

Relationship between macrophage colony-stimulating factor production by uterine epithelial cells and accumulation and distribution of macrophages in the uterus of pregnant mice.

Estrogen and progesterone induce production of macrophage colony-stimulating factor (CSF-1) by uterine epithelial cells, and CSF-1 is produced in the uterus during pregnancy in mice. CSF-1 is a lineage-specific stimulator of macrophage proliferation, chemotaxis, and function. High concentrations of macrophages accumulate in the uterus during pregnancy. Experiments were conducted to determine whether a relationship exists between intrauterine CSF-1 production and the number and distribution of uterine macrophages during pregnancy in mice. The study demonstrated that on day 1 of pregnancy CSF-1 bioactivity levels were high. The number of macrophages in the uterus was also high on days 1 and 2, and macrophages were concentrated at epithelial surfaces. The decrease in CSF-1 bioactivity seen between days 1 and 2 was followed by a decrease in the macrophage concentration. An increase in CSF-1 bioactivity on day 4 was followed by an increase in the concentration of intrauterine macrophages. During the immediate postimplantation period, macrophages were detected primarily in the myometrium and deep endometrium and CSF-1 bioactivity was undetectable. During the second half of pregnancy, when CSF-1 concentrations were very high, the macrophage concentration was also very high and large numbers of macrophages were detected in association with epithelia. The data confirmed the existence of a direct relationship between intrauterine CSF-1 and macrophage accumulation and suggested that macrophages are attracted to epithelial surfaces by CSF-1.

M-CSF (monocyte colony stimulating factor) and M-CSF receptor expression by breast tumour cells: M-CSF mediated recruitment of tumour infiltrating monocytes?

Infiltrating immune cells in 30 primary human epithelial breast tumours were studied using specific anti-CD3 (T cells), anti-CD68 (macrophages), anti-CD57 (NK cells), and an anti-pan-B cell antibody (L26). The majority of tumour infiltrating inflammatory cells are T cells (40-50%) and monocytes/macrophages (15-35%). The macrophage specific chemo-attractant and growth factor CSF-1 is detected by immunohistochemical techniques (IHC) at the level of invasive breast cancer cells in 46/50 tumours but not at the level of in-situ (pre-invasive) cancer. A mosaic staining pattern was usually observed, with a very high expression in areas of obvious stromal invasion (90% cells positive) and absent or trace staining in intraductal carcinoma. Macrophages and plasma cells are equally intensely positive. In-situ hybridisation experiments confirm the production of CSF-1 (mRNA) by tumour cells and show the same pattern of expression. Expression of the CSF-1 receptor protein (fms) was also observed by IHC in 41/48 invasive tumours, albeit at weaker intensities than in tumour infiltrating monocytes/macrophages. A concomitant expression of both CSF-1 and fms in in-situ carcinoma was never seen (n = 14). It is therefore proposed that the associated expression of CSF-1 and its receptor may be linked to the invasive potential of breast cancer, the monocytic infiltrate being an indication of the quantitative importance of CSF-1 production by the tumour.

Macrophage colony stimulating factor controls macrophage recruitment to the cycling mouse uterus

Progesterone and estrogen induce uterine epithelial cells to produce macrophage colony stimulating factor (CSF-1), and mouse uterine epithelial cells produce large amounts of CSF-1 during embryo development. The present study demonstrated that estrogen and progesterone each induce uterine CSF-1 gene transcription and translation detectable by Northern blotting and bioassay. Intrauterine CSF-1 production was greater in the presence of both estrogen and progesterone than following exposure to either hormone alone. CSF-1 mRNA was detectable in the uterus throughout the estrous cycle while CSF-1 bioactivity was detected only during proestrus. CSF-1 production was directly related to the presence of macrophages in the uterus. Ovariectomy, which was rapidly followed by a loss of uterine CSF-1 gene transcription, also was followed by a dramatic decrease in the number of uterine macrophages. Both uterine CSF-1 and uterine macrophages were reconstituted in a dose-dependent manner by systemic administration of estrogen or progesterone. High concentrations of circulating estrogen and progesterone increased the number of macrophages in the uterus and increased their accumulation near epithelial surfaces. Similar relationships were observed in the uterus of cycling mice. Macrophages accumulated in the uterus following intraluminal injection of recombinant human CSF-1 to ovariectomized mice, directly demonstrating the ability of CSF-1 to recruit macrophages from peripheral blood into the uterus. These studies demonstrated that estrogen and progesterone stimulation of CSF-1 production by mouse uterine epithelial cells controls recruitment and distribution of macrophages in the uterus during the estrous cycle.

Contributions of mesangial cells to glomerular immune functions

Mesangial cells play an important role in the maintenance of the structure and function of the glomerulus. In addition mesangial cells are capable of macromolecular uptake, and generation of autocoids and cytokines. Our studies have explored the mechanism of IgG uptake by cultured rat mesangial cells. Mesangial cells were found to express Fc receptors for IgG as demonstrated by specific binding studies, indirect immunofluorescence microscopy, immunoblotting and Northern blot analysis. The number of Fc receptors for IgG on mesangial cells was upregulated by interferon gamma, cyclic AMP and monocyte-macrophage specific colony stimulating factor--CSF-1. In addition uptake of IgG complexes was acutely increased by angiotensin II and inhibited by cAMP. These latter effects are independent of receptor number, but are mediated by changes in the cytoskeleton. These observations may be of significance for hemodynamically independent effects of vasoactive agents on mesangial function. Finally we examined the potential of mesangial cells for generation of CSF-1. Cultured mesangial cells produced radioimmunoassayable CSF-1 and expressed mRNA for CSF-1. Interferon gamma stimulated, whereas cAMP and agents increasing cAMP, such as forskolin and PGE2, inhibited CSF-1 production. This was due to decreased transcription resulting in lower levels of mRNA for CSF-1. The interactions of CSF-1, other cytokines, prostaglandins and cAMP may be important in regulating immune-like functions of mesangial cells, and especially their response to immunecomplexes.

Differential Production of Tumor Necrosis Factor, Macrophage Colony Stimulating Factor, and Interleukin 1 by Human Alveolar Macrophages

Human alveolar macrophages (AMO) have been investigated for their ability to produce three monokines, tumor necrosis factor-alpha (TNF), macrophage colony stimulating factor (CSF-1), and interleukin 1-beta (IL-1). No TNF activity was found in supernatants of unstimulated AMO cultured for 20 h, although TNF mRNA was detected in the cells by Northern blot analysis. Stimulation of the cells with lipopolysaccharide (LPS) induced production and release of high levels of TNF into the culture supernatant. Increased levels of TNF mRNA were detectable at 90 min after LPS stimulation by dot blot analysis, reaching maximum expression between 4 and 8 h and declining thereafter. TNF activity peaked at approximately 8 h in the AMO supernatants. After 24 h TNF production had ended. Compared to autologous monocytes the AMO produced 5.7 times more TNF on a per cell basis (activity accumulated in 20 h supernatants). Uncultured AMO expressed CSF-1 mRNA which was translated into active protein recovered in supernatants upon culture in the absence of stimulus. The addition of LPS to AMO slightly reduced both mRNA levels and amount of factor in the supernatants. In contrast to the AMO, monocyte production of CSF-1 was enhanced by LPS. CSF-1 production by AMO continued for at least 48 h of culture. Spontaneous production of low amounts of IL-1 was found in one-third of the AMO samples, while low levels of IL-1 mRNA were present in all tested preparations. LPS stimulation induced increase in IL-1 mRNA within 90 min; mRNA levels peaked between 12 and 20 h and stayed high for at least 42 h. However, while all LPS-stimulated AMO expressed high levels of IL-1 mRNA biologically active IL-1 was again detected only in a fraction of the AMO supernatants. These results show that the production of monokines CSF-1, TNF, and IL-1 is differentially regulated by LPS in alveolar macrophages and has different responses as compared to monocytes. The longevity of the messages for each of the factors is possible indicators of the relative contribution of these factors to the response to endotoxin-induced injury and repair processes in the lung.

Macrophage progenitor cell and colony-stimulating factor production during granulomatous schistosomiasis mansoni in mice.

Schistosoma mansoni worm eggs stimulate a T-cell-mediated granulomatous response in which macrophages play important inflammatory and regulatory roles. Although much has been learned about the functions of the schistosome granuloma macrophages, their origin and replicative ability are unknown. In the present sequential study, macrophage progenitor cells in the bone marrow (GM-CFC) and liver granulomas (M-CFC) were enumerated, and macrophage colony-stimulating factor (CSF-1) in the circulation and culture fluid of explanted granulomas of infected mice was assayed. During the acute phase of the infection, when the granulomatous response was vigorous (weeks 8 to 12) GM-CFC numbers were high in the bone marrow and M-CFC numbers were low within granulomas. Circulating CSF-1 levels were elevated, but the vigorous granuloma-secreted CSF-1 level was low. During the chronic phase of the infection, the number of GM-CFC within the bone marrow and levels of circulating CSF-1 returned to normal. Conversely, a sharp increase in the number of M-CFC occurred within the small immunomodulated granulomas that also secreted high levels of CSF-1. The frequency of M-CFC that proliferated under exogenously added CSF-1 within the immunomodulated granulomas was significantly higher than that of cells in the vigorous granulomas. Adherent macrophage-rich cells obtained after dispersal of the granulomas appeared to be one source of CSF-1 production. These data indicate that during the infection the macrophage supply to and replicative ability within the granulomas is influenced by systemically and/or locally produced CSF-1.

Regulation by interferon gamma of function in the acute monocytic leukemia cell line, THP-1.

THP-1 is an acute monocytic leukemia cell line which acquires phenotypic and functional monocytoid-like features following incubation with mezerein. The current study concerned the modulation of these features by rIFN gamma. rIFN gamma induces the time-dependent enhancement of HLA-DR expression in the presence or absence of mezerein but has no effect on the expression of Leu-M1, Leu-M2, or Leu-M3 antigens. CSF-1 production following mezerein activation was reduced by incubation in the presence of 10(3) and 10(4) units/ml rIFN gamma. This was confirmed through both biological assays with mouse bone marrow cells and an indirect ELISA. In contrast, the concentration of growth inhibitory activity in conditioned medium was increased by rIFN gamma. A small but significant increase in IL-1 beta concentration in conditioned medium was detected using a sensitive double-antibody ELISA and a radioimmunoassay. The results infer that the functional characteristics of this leukemia cell line are modulated by rIFN gamma in a manner qualitatively similar to that reported for IFN gamma treated normal monocytes.

Tumor Necrosis Factor (TNF)-alpha but not TNF-beta Induces Secretion of Colony Stimulating Factor for Macrophages (CSF-1) by Human Monocytes

Tumor necrosis factor (TNF)-alpha has been identified as a major inducer of colony stimulating factor (CSF)-secretion by human vascular endothelial cells and fibroblasts. In the present study we assessed the capacity of TNFs to induce release of CSF-1 from highly purified peripheral blood monocyte preparations. Whereas monocytes do not accumulate CSF-1 messenger (m)RNA constitutively and consequently do not produce CSF-1 protein, CSF-1 mRNA and protein secretion became detectable, when monocytes were cultured in the presence of TNF-alpha, that was synergistically enhanced by interferon-gamma (IFN-gamma). However, under identical experimental conditions TNF-beta failed to induce monocyte CSF-1 synthesis. Cultures of monocytes in the presence of TNF-beta before addition of TNF-alpha abolished the CSF-1 inducing capacity of TNF-alpha, suggesting that TNF-beta may act as antagonist to TNF-alpha for CSF-1 production. These data point out a previously unrecognized function of TNF-alpha to modulate CSF-1 release by monocytes and demonstrate disparate biological properties of different TNF species in hematopoiesis.

Tumor necrosis factor (TNF)-alpha but not TNF-beta induces secretion of colony stimulating factor for macrophages (CSF-1) by human monocytes

Tumor necrosis factor (TNF)-alpha has been identified as a major inducer of colony stimulating factor (CSF)-secretion by human vascular endothelial cells and fibroblasts. In the present study we assessed the capacity of TNFs to induce release of CSF-1 from highly purified peripheral blood monocyte preparations. Whereas monocytes do not accumulate CSF-1 messenger (m)RNA constitutively and consequently do not produce CSF-1 protein, CSF-1 mRNA and protein secretion became detectable, when monocytes were cultured in the presence of TNF-alpha, that was synergistically enhanced by interferon-gamma (IFN-gamma). However, under identical experimental conditions TNF-beta failed to induce monocyte CSF-1 synthesis. Cultures of monocytes in the presence of TNF-beta before addition of TNF-alpha abolished the CSF-1 inducing capacity of TNF-alpha, suggesting that TNF-beta may act as antagonist to TNF-alpha for CSF-1 production. These data point out a previously unrecognized function of TNF-alpha to modulate CSF-1 release by monocytes and demonstrate disparate biological properties of different TNF species in hematopoiesis.

Expression of the Macrophage-Specific Colony-Stimulating Factor in Human Monocytes Treated With Granulocyte-Macrophage Colony-Stimulating Factor

The macrophage-specific colony-stimulating factor (CSF-1, M-CSF) regulates the survival, growth and differentiation of monocytes. We have recently demonstrated that phorbol ester induces expression of CSF-1 in human monocytes. These findings suggested that activated monocytes are capable of producing their own lineage-specific CSF. The present studies demonstrate that the granulocyte-macrophage colony-stimulating factor (GM-CSF) also induces CSF-1 transcripts in monocytes. Furthermore, we demonstrate that the detection of CSF-1 RNA in GM-CSF-treated monocytes is associated with synthesis of the CSF-1 gene product. The results thus suggest that GM-CSF may indirectly control specific monocyte functions through the regulation of CSF-1 production. These findings indicate another level of interaction between T cells and monocytes.

Expression of the macrophage-specific colony-stimulating factor in human monocytes treated with granulocyte-macrophage colony-stimulating factor

The macrophage-specific colony-stimulating factor (CSF-1, M-CSF) regulates the survival, growth and differentiation of monocytes. We have recently demonstrated that phorbol ester induces expression of CSF- 1 in human monocytes. These findings suggested that activated monocytes are capable of producing their own lineage-specific CSF. The present studies demonstrate that the granulocyte-macrophage colony-stimulating factor (GM-CSF) also induces CSF-1 transcripts in monocytes. Furthermore, we demonstrate that the detection of CSF-1 RNA in GM-CSF- treated monocytes is associated with synthesis of the CSF-1 gene product. The results thus suggest that GM-CSF may indirectly control specific monocyte functions through the regulation of CSF-1 production. These findings indicate another level of interaction between T cells and monocytes.

Expression of the macrophage specific colony-stimulating factor (CSF-1) during human monocytic differentiation

We and others have previously demonstrated expression of the co-fms proto-oncogene during human monocytic differentiation. The c-fms gene has since been shown to encode for the macrophage specific colony stimulating factor (CSF-1) receptor. The present results demonstrate that both CSF-1 and c-fms transcripts are induced during monocytic differentiation of human HL-60 leukemia cells. The results further demonstrate that normal human monocytes express CSF-1 RNA and that the level of these transcripts increases upon treatment with phorbol ester. Finally, the detection of CSF-1 RNA in HL-60 cells and in monocytes is associated with production of the CSF-1 gene product. These findings would suggest that monocytes are capable of regulating their own survival, growth and differentiation through CSF-1 production.

Persistent production of colony-stimulating factor (CSF-1) by cloned bone marrow stromal cell line D2XRII after X-irradiation.

The adherent stromal layer in long-term bone marrow cultures (LTBMC) provides the cellular environment necessary for the in vitro proliferation and differentiation of pluripotential hematopoietic stem cells. The role of humoral hematopoietic growth factors, colony-stimulating factors (CSF) in the regulation of hematopoietic cell production in this system is poorly understood. We have recently isolated and cloned an adherent cell line, D2XRII, derived from murine LTBMC. Plateau phase 25 cm2 cultures of 2 X 10(6) D2XRII cells in 8.0 ml produced CSF-1 (M-CSF) at around 100-150 units/0.1 ml medium. Following X-irradiation there was a dose-dependent decrease in the production of CSF-1 to a plateau of 50% of control levels at 10,000 rad. Higher doses did not produce a further decrease. The X-ray dose reducing CSF-1 production to 50% was 100-fold above the lethal dose as measured by clonagenic survival following trypsinization and replating. Trypsinized replated viable adherent but nondividing X-irradiated D2XRII cells were maintained for up to 8 weeks after irradiation and demonstrated continuous production of CSF-1. The data indicate significant divergence of two biologic effects of X-irradiation on plateau-phase marrow stromal cells: physiologic function of adherence and CSF-1 production, versus proliferative integrity. This divergence of effects may be very relevant to understanding the mechanism of X-irradiation-associated marrow suppression and leukemogenesis.