Molecular Human Reproduction, Vol. 5, No. 5, 459-466,
May 1999
© 1999 European Society of Human Reproduction and Embryology
The expression, activity and regulation of granulocyte macrophage-colony stimulating factor in human endometrial epithelial and stromal cells*
Department of Obstetrics and Gynecology, University of Florida, Gainesville, FL 32610
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Abstract |
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The expression of granulocyte macrophage-stimulating factor (GM-CSF) and GM-CSF receptors in the human endometrium suggests an autocrine/paracrine role for GM-CSF in this tissue. Using primary cultures of isolated endometrial glandular epithelial and stromal cells, the present study examined: (i) the cell specific expression of GM-CSF and GM-CSF receptor mRNA and protein; (ii) direct action of GM-CSF on the rate of DNA synthesis and cell proliferation; and (iii) regulation of GM-CSF expression through its interaction with transforming growth factor (TGF)-ß1 in these cells. Quantitative reverse transcription–polymerase chain reaction (RT–PCR), enzyme-linked immunosorbent assay and immunocytochemistry indicates that glandular epithelial and stromal cells express GM-CSF, GM-CSF
and GM-CSF ß receptor mRNA and protein. The epithelial cells express a significantly higher level of GM-CSF mRNA than stromal cells while both types produce low concentrations of protein. At 0.01–100 ng/ml GM-CSF did not have a significant effect on the rate of [3H]-thymidine incorporation or proliferation of epithelial and stromal cells. However, GM-CSF (1 ng/ml) up-regulates its own protein expression, but does not effect TGF-ß1 mRNA protein expression in epithelial and stromal cells, and actually inhibits the cell-associated TGF-ß1 protein in stromal cells (P < 0.05). At 1 ng/ml TGF-ß1 up-regulates its own mRNA and protein expression in epithelial and stromal cells (P < 0.05), with no significant effect on GM-CSF expression. Co-treatment of the cells with GM-CSF + TGF-ß1 resulted in an increased production of GM-CSF protein as well as TGF-ß1 mRNA and protein expression by epithelial and stromal cells, compared with untreated controls (P < 0.001). In conclusion, the results suggest that GM-CSF is not a mitogenic factor for endometrial glandular epithelial and stromal cells, however, in an interactive manner with TGF-ß1 it regulates its own and the expression of TGF-ß1. epithelial cells/GM-CSF/human endometrium/stromal cells/TGF-ß
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Introduction |
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Cytokines are an integrated part of an interactive network of regulatory molecules which modulate various normal biological and pathological events effecting the endometrium (Chard, 1995
; Oral and Arici, 1996; Chegini and Williams, 1997 Chegini and Williams, 1999). Granulocyte macrophage-colony stimulating factor (GM-CSF) is a member of the haematopoietic cytokine family (Rasko and Gough, 1994) which is expressed in several reproductive tract tissues including endometrium of human and other species (Imakawa et al., 1993; Robertson et al., 1996; de Moraes et al., 1999). In the endometrium epithelial cells are the major site of GM-CSF expression (Imakawa et al., 1993, Robertson et al., 1996; Zhao and Chegini, 1999; de Moraes et al., 1999), while GM-CSF receptors are associated mostly with arteriole endothelial and stromal cells (Zhao and Chegini, 1999). Although GM-CSF is a major growth factor for granulocyte and macrophage proliferation, differentiation and survival, it also stimulates proliferation of several non-haematopoetic cell types (Rasko and Gough, 1994), including DNA synthesis, but not proliferation of mid-gestational trophoblasts (Drake and Head, 1994; Garcia-Loret et al., 1994; Jokhi et al., 1994a). The biological implication of GM-CSF in the human uterus is unknown, however, data obtained from animal models suggest that GM-CSF may play a key role in embryonic development throughout pregnancy and prevent spontaneous abortions in abortion-prone mice through a T cell-dependent mechanism involving inhibition of natural killer cell activity (Clark, 1993; Robertson et al., 1994). However, GM-CSF deficient mice are reported to be fertile (Dranoff et al., 1994).
Endometrial GM-CSF expression has been reported to be cycle-dependent, suggesting possible regulation by ovarian steroids (Robertson et al., 1996; Zhao and Chegini, 1999). In addition, other cytokines such as tumour necrosis factor (TNF)-, interleukin (IL-1) and transforming growth factor (TGF)-ß have been reported to regulate GM-CSF expression in several cell types including macrophages, fibroblasts and endothelial cells (Rasko and Gough, 1994). TGF-ß is a multifunctional cytokine which acts as an immunosuppressor factor by inhibiting multipotential haematopoietic cell growth, while enhancing the effect of other cytokines such as GM-CSF on the growth of more differentiated immune cells (Rasko and Gough, 1994), and preferentially stimulates GM-CSF-induced granulocyte over monocyte differentiation through a mechanism involving induction of GM-CSF receptors (Jacobsen et al., 1993). Human endometrium and isolated endometrial epithelial and stromal cells express TGF-ß isoform and TGF-ß receptor mRNA and protein, and TGF-ß isoforms were evaluated for their role in normal and pathological abnormalities affecting the reproductive tissues, including endometriosis and adhesion formation (Chegini et al., 1994a,b,c; Tang et al., 1994b; Juneja et al., 1996; Chegini, 1997; Chegini and Williams, 1997; Rong et al., 1997). Moreover, TGF-ß and GM-CSF has been shown to modulate the expression of integrins and extracellular matrix turnover which is an integrated part of the normal, as well as pathological events associated with the endometrium (Damsky et al., 1993; De-Nichilo and Burns, 1993; Van der Linden et al., 1994, 1995; Young and Lessey, 1997; Dou et al., 1999).
The objectives of the present study were to further investigate the role of GM-CSF in human endometrium by examining its expression and effect on isolated endometrial glandular epithelial and stromal cells and to determine the potential autocrine/paracrine action of GM-CSF and its interaction with TGF-ß1 in these cells.
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Materials and methods |
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All the materials and procedures used for isolation and culture of endometrial glandular epithelial and stromal cells, reverse transcription polymerase reaction (RT–PCR), construction of external cDNA standard template for quantitative RT–PCR, enzyme-linked immunosorbent assay (ELISA), immunocytochemistry, [3H]-thymidine incorporation and cell proliferation assays were as previously described (Tang et al., 1994b
; Zhao et al., 1995; Dou et al., 1996). Human specific GM-CSF and TGF-ß1 ELISA kits were purchased from Prospective Diagnosis Inc. (Cambridge, MA, USA) and Promega (Madison, WI, USA) respectively. A human T-lymphocyte cell line (HT-29) was purchased from American Type Culture Collection (Rockville, MD, USA).
The endometrial glandular epithelial and stromal cells were isolated from portions of uterine endometrial tissues obtained from premenopausal women aged 21–39 years, who were undergoing hysterectomy for medically indicated reasons (excluding endometrial cancer and leiomyomata), and were not under any hormonal treatments at the time of surgery. The collection of these tissues was approved by the Institutional Review Board. The endometrial glandular and stromal cells were isolated and cultured in Dulbecco's modified Eagle's medium (DMEM)–Ham's F-12 (50:50, v/v) supplemented with 10% fetal bovine serum (FBS) as previously described (Tang et al., 1994a,b). The glandular epithelial and stromal cells were routinely characterized using antibodies to cytokeratin, vimentin, desmin and smooth muscle actin, as previously described (Tang et al., 1994a,b).
Quantitative RT–PCR
To determine whether isolated endometrial glandular epithelial and stromal cells express GM-CSF and GM-CSF receptors and ß mRNA, the cells were cultured in 75 ml flask in the presence of 10% FBS until reaching ~90% confluent. Total cellular RNA was isolated from the cells and subjected to standard RT–PCR (Zhao et al., 1995). Restriction enzyme digestion with corresponding enzymes, amplification of cellular RNA without the reverse transcription step to detect the presence of any genomic DNA contamination, and tubes containing all the PCR components except the RT reaction mixture to check for the presence of DNA that may have carried over from a prior reaction, were used as controls.
For quantitative RT–PCR, cDNA was synthesized in a series of standard reactions each containing 2 µg of total RNA prepared from each cell types and several dilutions of competitive external cRNA standard (1x109 to 1x103 copies/µg of total RNA) as described (Dou et al., 1996, 1997a). The PCR products were separated on 2% agarose gels containing ethidium bromide and photographed, scanned and the band intensities were determined after their intensity values were normalized for their molecular weight (Dou et al., 1997b). The ratio of the band intensities within each lane were plotted against the copy number of added standard template/reaction and quantity of the target mRNAs were determined where the ratios of template/target band intensities were equal to one, and analysed by equations of best fit lines. The final number of mRNA molecules (copies/µg of total cellular RNA) was calculated as previously described (Dou et al., 1996, 1999).
Immunocytochemistry
To determine whether these cells synthesize GM-CSF and contain GM-CSF receptors, endometrial epithelial and stromal cells were cultured on 8-well Lab-Tek slides as previously described (Tang et al., 1994b). The cells were fixed and processed for immunocytochemical localization of GM-CSF, GM-CSF and GM-CSF ß receptors using their respective antibodies at 5 µg/ml prepared in phosphate-buffered saline (PBS), pH 7.4 containing 0.1% bovine serum albumin (BSA), and indirect immunofluorescence microscopy. Controls include incubation of the cell cultures with non-immune mouse or rabbit serum, or immunoglobulin G (IgG) instead of the primary antibodies.
Enzyme-linked immunoassay
To determine whether endometrial epithelial and stromal cells synthesize and release GM-CSF, the cells were cultured in 24-well dishes as above in the presence of 10% FBS for 48 h. The cells were washed with serum free media and further incubated in the presence of 2% FBS for an additional 24 h. The culture conditioned media from these cells (control) and cells treated with 1 ng/ml of GM-CSF, TGF-ß1 and GM-CSF + TGF-ß1 were collected, centrifuged, and stored at –80°C until assayed. Culture-conditioned media and medium unexposed to cells were assayed for GM-CSF and TGF-ß1 using ELISA according to the manufacturer's recommended procedure, with limit of detection of 2.5 and 2 pg/ml respectively (Rong et al., 1997).
[3H]-thymidine incorporation and cell proliferation assay
To determine whether GM-CSF is a mitogen for endometrial glandular epithelial and stromal cells, the cells were cultured in 96-well microplates at density of ~2.5x103 cells/well, in the presence of 10% FBS for 48 h (Tang et al., 1994b). The cells were washed, made quiescent in serum-free condition for 48 h, and the quiescent cells were then incubated in medium supplemented with 2% FBS in the presence and absence of various doses of recombinant human GM-CSF and 2 µCi/ml [3H]-thymidine to determine the rate of DNA synthesis after 48 h of incubation as described previously (Tang et al., 1994a). In another sets of experiments performed under the same condition without addition of [3H]-thymidine the rate of cell proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Tang et al., 1994a). HT29 T cells were cultured as previously described and used as a positive control (Zhao and Chegini, 1994).
Regulation of GM-CSF expression
Endometrial epithelial and stromal cells express GM-CSF and TGF-ß1 and receptors mRNA and protein, suggesting an autocrine or paracrine role for these cytokines in the endometrium. To investigate whether GM-CSF has any regulatory effect on its own expression and the possibility of interacting with TGF-ß1, quiescent endometrial epithelial and stromal cells were incubated with 2% FBS supplemented medium in the presence and absence of 1 ng/ml recombinant human GM-CSF, TGF-ß1 or GM-CSF + TGF-ß1 for 24 h. After the incubation conditioned media were collected, the cells were washed, scraped and used either for isolation of total cellular RNA, or lysed to obtain the total cellular proteins (Tang et al., 1994b). The concentrations of GM-CSF and TGF-ß1 (active and total) in the media, and TGF-ß1 in the cell lysate were determined by ELISA, and their mRNA expression by quantitative RT–PCR as described above.
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GM-CSF and GM-CSF receptors mRNA and protein expression
Standard RT–PCR indicated that isolated endometrial glandular epithelial and stromal cells maintained in primary culture express GM-CSF and GM-CSF
and ß receptor mRNA (Figure 1A). Digestion of PCR products with respective restriction enzymes resulted in the anticipated smaller fragments as shown for epithelial cells (Figure 1B), while, amplification without the reverse transcription step to detect any contaminating genomic DNA, and tubes containing all the PCR components except the RT reaction mixture to check for the presence of DNA that may have carried over from a prior reaction, were negative (not shown). These cells also contain immunoreactive GM-CSF, and GM-CSF and ß receptor proteins (Figure 2). In controls deletion or replacement of primary antibody with normal IgG (Figure 2G and H for GM-CSF), resulted in a lack of immunostaining over the epithelial and stromal cells (not shown). Quantitative RT–PCR of total RNA and protein isolated from epithelial and stromal cells indicated that they express ~4.6x104 and 1.5x104 copies of GM-CSF mRNA/µg of total cellular RNA, and 25 and 12 pg/ml of GM-CSF protein/106 cells respectively.
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The effect of GM-CSF on [3H]-thymidine incorporation and cell proliferation
GM-CSF at concentrations of 0.1–100 ng/ml did not have any effect on the rate of [3H]-thymidine incorporation or cell proliferation of serum-deprived quiescent endometrial glandular epithelial and stromal cells (data not shown). Quiescent epithelial and stromal cells can be stimulated to enter cell cycle in the presence of 10% FBS, or half-stimulated with 2% FBS (Tang et al., 1994a
). Treatment of half-stimulated epithelial and stromal cells with GM-CSF at 0.1–100 ng/ml also had no significant effect on rate of [3H]-thymidine incorporation, or cell proliferation compared to controls (Figure 3A and B). However, GM-CSF significantly increased the rate of [3H]-thymidine incorporation and proliferation of HT-29 T lymphocytes used as positive control (P < 0.05, Figure 3C).
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–Interaction between GM-CSF and TGF-ß1
Since glandular epithelial and stromal cells express GM-CSF, TGF-ß and their receptors, we further examined possible self-regulation and interactions between these cytokines in epithelial and stromal cells. The level of GM-CSF mRNA expression in epithelial and stromal cells was not altered after treatment with GM-CSF, TGF-ß1 or GM-CSF + TGF-ß1 (1 ng/ml) compared to untreated control (data not shown). However, GM-CSF protein production was significantly increased after treatment with GM-CSF in both cells compared to TGF-ß1 -treated and untreated control (P < 0.05), with a slight increase after GM-CSF + TGF-ß1 treatment in epithelial cells (Figure 4A and D
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) and total (active + latent (
) TGF-ß1 production by endometrial epithelial (A–C) and stromal (D–F) cells incubated in the presence of 2% fetal bovine serum (FBS) for 24 h. A, B, D and E show the levels of GM-CSF and TGF-ß1 released into the culture condition media, and C and F show the cell-associated TGF-ß1. The cells are derived from endometrial tissues of early secretory phase of the menstrual cycle. Points represent mean ± SEM of triplicate experiments using isolated cells from three uteri. *Significantly different from controls.The endometrial epithelial and stromal cells express 2.4 ± 0.3x105 and 0.9 ± 0.1x105 copies of TGF-ß1 mRNA/µg total cellular RNA; this was significantly up-regulated following treatment of these cells with TGF-ß1 at 1 ng/ml concentration to 5.2 ± 0.7x106 and 2.3 ± 0.2 106 copies respectively (P < 0.05, Figure 5
). The expression of TGF-ß1 mRNA was also enhanced in response to GM-CSF treatment of epithelial (4.1 ± 0.3x105 copies) and stromal (9.8 ± 1.8x105 copies) cells as well as in response to TGF-ß1 + GM-CSF treatments (5.4 ± 0.7x106 and 3.2 ± 0.7x106 copies) compared with GM-CSF-treated, and untreated controls (P < 0.05 Figure 5). At the protein level, the untreated epithelial and stromal cells synthesized and released <1 ng/ml of total TGF-ß1 of which <400 pg/ml was in an active form, however, a major portion of TGF-ß1 was found to be cell-associated accounting for >20 ng/ml of total and >2 ng/ml of active TGF-ß1 (Figure 4B, C, E and F
). Treatment with GM-CSF decreased, while treatment with TGF-ß1 significantly increased, the level of total and active TGF-ß1 production released into epithelial and stromal cell culture-conditioned media (Figure 4B and E). In comparison with treatment using TGF-ß1 alone, co-treatment of the cells with GM-CSF + TGF-ß1 significantly inhibited the production of both total and active TGF-ß1 by epithelial (P < 0.05), but not stromal cells (Figure 4B and E). In regard to cell-associated TGF-ß1, treatment with GM-CSF had no effect on TGF-ß1 synthesis by epithelial cells (Figure 4C), whereas significantly inhibited the stromal cell associated total TGF-ß1 synthesis (P < 0.05; Figure 4F). Treatment with TGF-ß1 increased the level of cell-associated TGF-ß1 in both cell types with significantly higher active TGF-ß1 in epithelial cells and total TGF-ß1 in stromal cells (P < 0.05). Co-treatment with GM-CSF + TGF-ß1 resulted in inhibition of cell-associated total TGF-ß1 in both cells, compared with TGF-ß1-treated cells (Figure 4C and F).
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Discussion |
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We have demonstrated that human endometrium expresses GM-CSF and GM-CSF receptors mRNA and protein, with luminal/glandular epithelial cells as the primary site of GM-CSF expression (Zhao and Chegini, 1999
). We have now extended these observations and, in this study, have demonstrated that isolated endometrial glandular epithelial and stromal cells maintained in primary culture continue to express GM-CSF and GM-CSF receptor mRNA and protein. As expected the epithelial cells express a significantly higher level of GM-CSF mRNA than the stromal cells, however, both cell types released a very low level of GM-CSF protein into their culture conditioned media, despite the presence of relatively strong immunoreactive GM-CSF in both cell types. While our observations suggest that GM-CSF is mostly cell-associated, a previous bioassay study has reported that primary cultures of endometrial epithelial cells release as much as 4.4 ng/ml of GM-CSF into their culture conditioned media (Giacomini et al., 1995). The discrepancy between this and our study may be due to culture conditions as well as an inherent limitation of the bioassay in which other autocrine factors released into the culture conditioned media can, in an additive or synergistic manner with GM-CSF, stimulate the T lymphocyte proliferation used in the bioassay.
In this regard, cytokines such as TNF- and IL-1 which are expressed by human endometrium have been shown to induce GM-CSF expression in macrophages, fibroblasts and endothelial cells (Rasko and Gough, 1994). In the present study we demonstrated that GM-CSF up-regulates its own expression at protein level in endometrial epithelial and stromal cells and that its effect is enhanced after co-treatment with TGF-ß1 in epithelial, but not stromal cells. Because endometrial epithelial and stromal cells express TGF-ß1 mRNA and protein (Tang et al., 1994b), we further demonstrated that TGF-ß1 also up-regulates its own expression at the mRNA and protein levels in these cells. Interestingly, GM-CSF up-regulates the expression of TGF-ß1 at the mRNA level, but inhibits TGF-ß protein production by epithelial and stromal cells. Such an inhibitory action of GM-CSF on TGF-ß1 production was limited only to epithelial cells after co-treatment with TGF-ß1, while cell-associated TGF-ß1 was inhibited by GM-CSF in stromal cells, and by GM-CSF + TGF-ß1 in both cell types. Collectively, the data suggest that effect of GM-CSF, TGF-ß1 and their interactions on GM-CSF and TGF-ß1 expression occur at different regulatory- and compartmental-specific manner. Considering the compartmental-specific expression of TGF-ß1, GM-CSF and their receptors in the endometrium, such interactions may also result in a differential regulation of their expression at tissue level.
The interactions between TGF-ß and GM-CSF, as well as other cytokines, have been well documented in several cell types. For instance, TGF-ß1 has been shown to inhibit TNF-, IL-1, IL-2, IL-3 and interferon (IFN)-ß expression which are assumed to implement TGF-ß-induced immunosuppressive action (Rosko and Gough, 1994). TGF-ß has also been shown to inhibit IL-1- and TNF--induced GM-CSF production in chondrocytes (Alsalameh et al., 1994), GM-CSF-induced Ia gene expression in bone marrow macrophages primarily through post-transcriptional regulation (Gollnick et al., 1995), and IL-3, IL-9, IL-11 and GM-CSF-induced CD34+ cell total colony formation (Lemoli et al., 1995). TNF--induced IL-1 and GM-CSF production by endothelial cells and fibroblasts has been suggested to constitute an important step in the inflammatory process (Rasko and Gough, 1994), as anti-inflammatory agents in general suppress cytokine-induced GM-CSF production (Alsalameh et al., 1994). TGF-ß has also been shown to inhibit IFN-
, TNF- and GM-CSF production by the human first trimester uterine lymphocyte, population of decidual CD56+ NK cells which are the major site of GM-CSF and TGF-ß2-like protein production (Clark, 1993; Jokhi et al., 1994a; Blone et al., 1995), and IL-5, IL-3, GM-CSF and IFN--induced eosinophil (CD16) survival, which was completely neutralized with anti-TGF-ß antibody (Atsuta et al., 1995). However, GM-CSF antagonizes TGF-ß-induced expression of Fc RIII in peripheral blood monocytes by suppressing TGF-ß mRNA expression and signalling pathway involving tyrosine kinase activity (Kruger et al., 1996). In contrast, TGF-ß has been shown to enhance the expression of GM-CSF receptors in murine peripheral macrophages, suggesting a possible mechanism for the synergistic interaction with TGF-ß (Fan et al., 1992). Considering that these cytokines are expressed in human endometrium, we propose the existence of delicate feedback interactions between endometrial-derived GM-CSF and TGF-ß with potential ability to interact with other endometrial-derived cytokines.
GM-CSF which is a mitogen for haematopoietic and several non-haemopoetic cell types such as osteoblasts, smooth muscle, endothelial and epithelial cells (Rasko and Gough, 1994), did not appear to have any effect on the proliferation of endometrial glandular epithelial and stromal cells. GM-CSF has been shown to stimulate the rate of DNA synthesis, but not proliferation of mid-gestational trophoblasts (Drake and Head, 1994; Garcia-Loret et al., 1994; Jokhi et al., 1994a), the development of mouse blastocysts, and ovine embryonic trophectoderm IFN-
production which is considered important in maternal recognition of pregnancy in this species (Clark, 1993; Imakawa et al., 1993; Robertson et al., 1994). For these reasons, GM-CSF is considered a key factor in embryonic development throughout the pregnancy, although mice lacking GM-CSF are fertile (Dranoff et al., 1994). In women with endometriosis and peritoneal adhesions (conditions which are often associated with peritoneal inflammation), the peritoneal fluid concentrations of TGF-ß1, but not GM-CSF, have been shown to be significantly higher than in normal subjects (Oosterlynck et al., 1995; Chegini et al., 1999). This has also been reported in surgically-included adhesion formation in mice (Rong et al., 1997). However, GM-CSF over-expression through gene transfer into the lung has been shown to induce granulation tissue formation, which was associated with a marked increase in the TGF-ß1 content of bronchoalveolar lavage fluid, with peak production coinciding with the emergence of -smooth muscle actin-rich myofibroblasts (Xing et al., 1997). Macrophages purified from bronchoalveolar lavage fluid after GM-CSF gene transfer spontaneously released significant quantities of TGF-ß1 protein in vitro (Xing et al., 1997). The products of activated peritoneal macrophages and lymphocytes are reported to play a key role in the pathogenesis of endometriosis and adhesion formation (Hill, 1992; Juneja et al., 1995, 1996; Chegini, 1997; Chegini and Williams, 1997). GM-CSF and TGF-ß have also been shown to regulate the expression of integrin v in cultured human macrophages (De-Nichilo and Burns, 1993), and other integrins in a human macrophage cell line (Dou et al., 1999). The stage-specific expression of integrins, such as v and ß3 in human endometrium, has been suggested to participate in endometrial receptivity for embryo implantation, and their absence in subjects with endometriosis-associated infertility (Van der Linden et al., 1995; Young and Lessey, 1997). If GM-CSF and TGF-ß have a regulatory effect on endometrial integrins expression, it further implicates their potential role in various endometrial activities including preparation for embryo implantation.
In addition to GM-CSF regulation by other cytokines, the apparent menstrual cycle variations in endometrial GM-CSF expression suggests possible involvement of the ovarian steroids in its regulation (Zhao and Chegini, 1999), although, GM-CSF production by isolated endometrial glandular epithelial cells was not dependent on the phase of the menstrual cycle of the tissues (Giacomini et al., 1995). Our preliminary data, however, indicate that oestradiol may stimulate GM-CSF production by endometrial stromal cells (unpublished data). Similarly, treatment of mouse uterine epithelial cells with oestradiol has been reported to stimulate GM-CSF expression while progesterone, which did not have any affect alone, inhibited the action of oestradiol (Robertson et al., 1996).
In conclusion, we provided further evidence that endometrial epithelial and stromal cells express GM-CSF and GM-CSF receptors. Although it was found not to be a mitogen for these cells, GM-CSF either alone or through interaction with TGF-ß regulates its own and TGF-ß expression. Considering that cytokines play a key role in various endometrial biological activities, such regulatory interactions are critical to maintain their optimal expression and the normal endometrial biological functions. Disorderly expression of one or more of these factors may also alter the endometrial molecular environment leading to abnormalities such as endometriosis.
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Notes |
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*Presented in part at the 42nd annual meeting of Society for Gynecologic Investigation, March, 1995, Philadelphia, PA, USA.
1 To whom correspondence should be addressed 
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Submitted on April 24, 1998; accepted on February 15, 1999.
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