Citations of BMS242
The value of a single combined measurement of VEGF, glycodelin, progesterone, PAPP-A, HPL and LIF for differentiating between ectopic and abnormal intrauterine pregnancy
Daponte,A.; Pournaras,S.; Zintzaras,E.; Kallitsaris,A.; Lialios,G.; Maniatis,A.N.; Messinis,I.E.
Human Reproduction 2005;20:3163-3166.
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Abstract
BACKGROUND: To evaluate whether serum concentrations of the non-placental markers vascular endothelial growth factor (VEGF), glycodelin (GLY) and progesterone (P) and the novel placental markers pregnancy-associated plasmaprotein A (PAPP-A), human placental lactogen (HPL) and leukaemia inhibiting factor (LIF) differ in ectopic pregnancy (EP) when compared with abnormal intrauterine pregnancy (aIUP). METHODS: A prospective clinical study was conducted at the University Hospital of Larissa, Greece. The study included 50 patients admitted with failed pregnancy and suspected ectopic pregnancy that were treated with curettage or laparoscopy and classified as histologically confirmed EPs (n = 27) or histologically confirmed aIUPs (n = 21) (mean gestational age of 7.15 and 7.3 weeks, respectively). Two suspected EPs proved to be normal IUPs and were excluded. VEGF, GLY, P, {beta}-HCG, PAPP-A, HPL and LIF were measured by enxyme-linked immunosorbent assay (ELISA) methods in a single pre-operative blood sample. RESULTS: The median VEGF concentration was 227.2 pg/ml in the EP group versus 107.2 pg/ml in the aIUP group (P < 0.001), with a suggested threshold value of 174 pg/ml for their differential diagnosis. LIF, P, PAPP-A, HPL and GLY serum measurements did not differ significantly between EP and aIUP. CONCLUSION: VEGF serum levels might be a useful marker in differentiating between EPs and aIUPs
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Nonapoptotic effects of tumor necrosis factor-related apoptosis-inducing ligand on interleukin-6, leukemia inhibitory factor, interleukin-8, and monocyte chemoattractant protein 1 vary between undifferentiated and decidualized human endometrial stromal cells
Fluhr,H.; Sauter,G.; Steinmuller,F.; Licht,P.; Zygmunt,M.
Fertil.Steril. 2009;› Link
Abstract
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), which is known to exert death-inducing as well as nonapoptotic functions, has been shown to be expressed by the early trophoblast. Here we report that TRAIL has no apoptotic effects on human endometrial stromal cells, but differentially regulates cytokines and chemokines and might therefore play a role in the modulation of the cytokine milieu at the implantation site
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Polarized secretion of Leukemia Inhibitory Factor
Hill,Eric J.; Vernallis,Ann B.
BMC Cell Biology 2008;9:53› Link
Abstract
The direction of cytokine secretion from polarized cells determines the cytokine's cellular targets. Leukemia inhibitory factor (LIF) belongs to the interleukin-6 (IL-6) family of cytokines and signals through LIFR/gp130. Three factors which may regulate the direction of LIF secretion were studied: the site of stimulation, signal peptides, and expression levels. Stimulation with IL-1¦ is known to promote IL-6 secretion from the stimulated membrane (apical or basolateral) in the human intestinal epithelial cell line Caco-2. Since LIF is related to IL-6, LIF secretion was also tested in Caco-2 following IL-1¦ stimulation. Signal peptides may influence the trafficking of LIF. Two isoforms of murine LIF, LIF-M and LIF-D, encode different signal peptides which have been associated with different locations of the mature protein in fibroblasts. To determine the effect of the signal peptides on LIF secretion, secretion levels were compared in Madin-Darby canine kidney (MDCK) clones which expressed murine LIF-M or LIF-D or human LIF under the control of an inducible promoter. Low and high levels of LIF expression were also compared since saturation of the apical or basolateral route would reveal specific transporters for LIF. Results When Caco-2 was grown on permeable supports, LIF was secreted constitutively with around 40% secreted into the apical chamber. Stimulation with IL-1¦ increased LIF production. After treating the apical surface with IL-1¦, the percentage secreted apically remained similar to the untreated, whereas, when the cells were stimulated at the basolateral surface only 20% was secreted apically. In MDCK cells, an endogenous LIF-like protein was detected entirely in the apical compartment. The two mLIF isoforms showed no difference in their secretion patterns in MDCK. Interestingly, about 70% of murine and human LIF was secreted apically from MDCK over a 400-fold range of expression levels within clones and a 200,000-fold range across clones. Conclusion The site of stimulation affected the polarity of LIF secretion, while, signal peptides and expression levels did not. Exogenous LIF is transported in MDCK without readily saturated steps
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Leukaemia inhibitory factor and interleukin 11 levels in uterine flushings of infertile patients with endometriosis
Mikolajczyk,M.; Wirstlein,P.; Skrzypczak,J.
Human Reproduction 2006;21:3054-3058.
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Abstract
BACKGROUND: Exact aetiology of infertility in stage I/II endometriosis patients is not known. Interleukin 11 (IL-11) and leukaemia-inhibitory factor (LIF) are factors associated with implantation window in human eutopic endometrium. We decided to test whether there is an altered secretion of these factors, which could explain receptivity defect in patients with minimal endometriosis. METHODS: Uterine flushing and endometrial samples were collected 7-9 days after ovulation (implantation window) from infertile patients with stage I/II endometriosis (n = 14) and fertile, endometriosis-free controls (n = 21). IL-11 and LIF were assessed in uterine flushings in eutopic endometria in all patients by enzyme-linked immunosorbent assay (ELISA). In eutopic endometrium, semiquantitative RT-PCR was performed for LIF and IL-11 mRNA expressions. RESULTS: No statistically significant differences were found in uterine flushing in women with and without endometriosis with regard to IL-11 levels (0.0 pg/ml versus 0.0 pg/ml) and LIF (25.53 pg/ml versus 36.26 pg/ml). These results were confirmed by the results of RT-PCR, where there were also no differences between studied groups. CONCLUSIONS: There is no receptivity defect with regard to LIF and IL-11 secretions by eutopic endometrium in infertile women with endometriosis
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Soluble gp130 is Up-Regulated in the Implantation Window and Shows Altered Secretion in Patients with Primary Unexplained Infertility
Sherwin,J.R.A.; Smith,S.K.; Wilson,A.; Sharkey,A.M.
Journal of Clinical Endocrinology & Metabolism 2002;87:3953-3960.
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Abstract
Members of the IL-6 family of cytokines, which includes leukemia inhibitory factor (LIF) and IL-11, play important roles in implantation. The activity of these cytokines is modified by soluble receptors such as the IL-6 receptor (sIL-6R). gp130 is a signal transduction molecule common to the receptor complexes of this family, and its soluble form (sgp130) antagonizes their actions. The purpose of this study was to determine whether secretion of IL-6, LIF, sIL-6R, and sgp130 was different in the endometrium of women with primary unexplained infertility compared with normal fertile women. Endometrial biopsies were taken between d LH+6 and +13 and cultured in serum-free medium for 4 h. Secretion of IL-6, LIF, sIL-6R, and sgp130 was measured in the supernatant by ELISA. We also measured the secretion of IL-6, sIL-6R, and sgp130 by endometrial biopsies taken throughout the menstrual cycle in normal fertile women. Secretion of sgp130 increased 20-fold between d 20 and 26 of the cycle, coinciding with the implantation window (proliferative phase, median, 27.0 pg/ml{middle dot}mg; range, 23-36; d 20-26, median, 501.5 pg/ml{middle dot}mg; range, 26.1-1344; P = 0.03). RT-PCR showed that none of the known splice variants of gp130 were present in endometrium, indicating that sgp130 is produced by proteolytic cleavage of the membrane-bound form. IL-6 secretion varied considerably between patients and was greatest during the secretory phase and at menstruation. No significant change was seen in sIL-6R during the cycle. Between LH+6 and +13, secretion of sgp130 was significantly reduced in the infertile group (median, 93.1 pg/ml{middle dot}mg; range, 28.5-256; compared with the fertile group, median, 223 pg/ml{middle dot}mg; range, 63-534; U-statistic = 37; P = 0.017). Secretion of IL-6, LIF, and sIL-6R did not differ between the two groups. Immunolocalization of gp130, IL-6R, and the LIF receptor showed that the glandular epithelium and also endothelial cells are targets for IL-6 and LIF. These findings show that during a normal menstrual cycle, sgp130 secretion is greatly increased between d LH+6 and +13, due to proteolytic cleavage of membrane-bound gp130. Infertile patients show reduced secretion of sgp130 compared with fertile controls during this period, which coincides with the implantation window
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The human hepatocyte growth factor (HGF) gene is transcriptionally activated by leukemia inhibitory factor through the Stat binding element
Tomida,Mikio; Saito,Takeshi
Oncogene 2003;23:679-686.
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Abstract
We found that human melanoma SEKI and neuroepithelioma NAGAI cells, which are known to secrete high concentrations of leukemia inhibitory factor (LIF), also secrete high levels of hepatocyte growth factor (HGF). We therefore examined the role of LIF in HGF expression and examined the human HGF promoter. The expression of both LIF and HGF mRNA is very low in HEK293 cells. Treatment of these cells with LIF stimulated the expression of HGF mRNA. The cis-acting regulatory element of the HGF promoter in SEKI and 293 cells was analysed by means of a transient expression assay. By deletion analysis, we showed that the region comprising the -181 to -73 bp was required for full activity of the HGF promoter in SEKI cells and for LIF responsiveness of 293 cells. This region contains putative consensus sequences for the Stat and NF-IL6 (C/EBP beta) transcription factors. The activity of the HGF promoter was abolished by mutation of the Stat site at -99/-91, while the activity only slightly decreased on mutation of the NF-IL6 site. Treatment with anti-LIF antibodies or interruption of Stat3 signaling by dominant-negative Stat3 also reduced the HGF promoter activity. Stat3 activation was constitutive in SEKI cells and induced on treatment of 293 cells with LIF. These results suggest that cytokines, growth factors and oncogenes (v-Src, etc.) that activate Stat3 are important regulators of HGF expression.
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Effects of IFN-@b, leptin and simvastatin on LIF secretion by T lymphocytes of MS patients and healthy controls
Vanderlocht,J.; Hendriks,J.J.A.; Venken,K.; Stinissen,P.; Hellings,N.
Journal of Neuroimmunology 2006;177:189-200.
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Abstract
In multiple sclerosis (MS), oligodendrocyte injury is believed to be caused by an aberrant immune response initiated by autoreactive T cells. Increasing evidence indicates that inflammatory responses in the central nervous system are not exclusively detrimental, but may also exert protective effects. Such protective effects are potentially mediated by the local secretion of neurotrophic factors by immune cells. We previously reported that T cells and monocytes in vitro and in inflammatory MS lesions produce leukaemia inhibitory factor (LIF), a member of the neuropoietic family of neurotrophins. In the present study, we report a reduced LIF production by CD4+ T cells of relapsing remitting MS patients as compared to healthy controls. Furthermore, immunomodulatory agents such as leptin, IFN-ß and simvastatin were studied for their potential to alter LIF and secretion of other cytokines by T cells and monocytes of relapsing remitting MS patients and healthy controls. Low doses of simvastatin, but not IFN-ß or leptin enhanced LIF secretion by CD4+ T cells of RR-MS patients. We further demonstrated that LIF did not influence viability, proliferation and cytokine secretion of T cells. Together these data provide new information on the regulation of LIF secretion by immune cells. Further insights into the complex regulation of neurotrophic factors such as LIF may prove useful for treatment of MS.
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Nucleofection Is a Valuable Transfection Method for Transient and Stable Transgene Expression in Adipose Tissue-Derived Stem Cells
Zaragosi,Laure Emmanuelle; Billon,Nathalie; Ailhaud,Gerard; Dani,Christian
Stem Cells 2007;25:790-797.
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Abstract
Adipose tissue-derived stem cells are a powerful tool for in vitro study of adult stem cell biology. So far, they have not been extensively used for gain or loss of function studies since they are resistant to most common transfection methods. Herein, we tested several classic transfection methods on human multipotent adipose tissue-derived stem (hMADS) cells. Our results showed that lipofectants and calcium phosphate were poorly efficient for transgene delivery in hMADS cells. In contrast, nucleofection, an electroporation-based method that is assumed to target plasmid DNA directly to the cell nucleus, led to a significant transient transgene expression in hMADS cells (up to 76% enhanced green fluorescent protein [EGFP]-positive cells were detected). Furthermore, after selection of hMADS cells that were nucleofected with a selectable plasmid coding for EGFP, stable EGFP expressing clones could be propagated in culture and efficiently induced to differentiate into EGFP-positive adipocytes and osteoblasts. Finally, we verified that nucleofected hMADS cells could produce a functional, transgene-encoded, secreted protein. To this aim, hMADS cells were nucleofected with a plasmid coding for leukemia inhibitory factor (LIF). This protein was detected at high concentrations in supernatants from pCAG-LIF transfected hMADS cells. Moreover, supernatants were able to maintain mouse embryonic stem cells' undifferentiated phenotype, indicating that hMADS cells could secrete a functional LIF protein. Taken together, our data demonstrate that nucleofection allows both transient and stable gene expression in adipose tissue-derived stem cells, without impairing their differentiation potential
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