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Interleukin 1 beta is induced by interleukin 11 during decidualization of human endometrial stromal cells, but is not released in a bioactive form
Abstracts / Journal of Reproductive Immunology 71 (2006) 135–176
We can correlate comprehensive ultrasonographic data with various and distinct mechanisms that were described in animal models as being involved in implantation failure. Keywords: Cytokines; Implantation; Implantation; Vascularisation doi:10.1016/j.jri.2006.08.019 [O4] Interleukin 1 beta is induced by interleukin 11 during decidualization of human endometrial stromal cells, but is not released in a bioactive form C.A. White a,b, , E. Dimitriadis a , A.M. Sharkey c , C. Stoikos a , L. Salamonsen a
147
release. Bioactive IL1B was detected in vivo at very low levels and at discrete foci in late secretory phase and first trimester decidua. This regulation of latent and bioactive IL1B at the fetal–maternal interface may prime decidual cells to respond rapidly to immunological challenge or to signals from the blastocyst during implantation.
Reference White, C.A., Dimitriadis, E., Sharkey, A.M., Salamonsen, L.A., 2005. Interleukin-11 inhibits expression of insulin-like growth factor binding protein-5 mRNA in decidualizing human endometrial stromal cells. Mol Hum Reprod 11, 649–658.
Keywords: Cytokines; Chemokines and signalling; Decidua; Implantation
a
Prince Henry’s Institute of Medical Research, Australia; b Monash University, Australia; c University of Cambridge, UK Blastocyst implantation is dependent on the differentiation of endometrial stromal cells (ESC) into decidual cells. Decidualization of human ESC in vitro is enhanced by interleukin 11 (IL11), with associated changes in gene expression. Our previous microarray study identified interleukin 1 beta (IL1B) as the most upregulated gene during IL11-enhanced decidualization [1]. This study aimed to examine the effect of IL11 on interleukin 1 beta (IL1B) mRNA and protein expression during in vitro decidualization, and determine the functional significance of increased IL1B levels during this process. Stromal cells were isolated from endometrial biopsies (n = 21) obtained between days 7 and 26 of the menstrual cycle from women with no known endometrial dysfunction. Cells were decidualized with 17-estradiol and medroxyprogesterone acetate in the presence or absence of exogenous IL11, and IL1B mRNA was quantified by real-time RT–PCR. Inactive proIL1B and bioactive IL1B in cell lysates and conditioned media were measured using specific immunoassays. Secretion of bioactive IL1B from decidualizing ESC was investigated by in vitro stimulation with lipopolysaccharide, interferon gamma or human chorionic gonadotropin. Immunostaining was carried out on in vitro decidualized ESC and cycling and pregnant decidua using antibodies specific for total and bioactive IL1B. Consistent with the microarray analysis, exogenous IL11 increased the abundance of IL1B mRNA in decidualizing ESC by 28-fold (range 1.3–107.1-fold), and total immunoreactive IL1B was also increased. However, this was not reflected in bioactive IL1B secretion from these cells, and none of the tested stimuli were able to induce its
doi:10.1016/j.jri.2006.08.020 [O5] Role of signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signalling 3 (SOCS3) in regulation of trophoblast invasion T.G. Poehlmann a, , S. Bachmann b , I. Rudloff a , J. R¨odiger a , C. Neblung a , E. Schleussner a a
Friedrich-Schiller-Universit¨at Jena, Germany; f¨ur Herz- und Lungenforschung, Germany b Max-Planck-Institut
Invasion of trophoblast cells requires a fine tuning, which is fundamental for correct placentation. This tuning is regulated on intracellular level. Several signal transducers and their suppressors, known from former studies and tumor invasion are expected to be involved. The activation of the Signal Transducer and Activator of Transcription (STAT)-3 correlates with the invasiveness of carcinoma and trophoblast cells. Its negative regulator, Suppressor of Cytokine Signalling (SOCS)-3 seems to be highly expressed in non invasive trophoblasts from sheep placentomae. Aim of this investigation was to Knock Down STAT3 and SOCS3 by siRNA and shRNA in human trophoblast and carcinoma cells in order to verify the role of these molecules during human placentation. siRNA was applied to choriocarcinoma cells to knock down STAT3 and SOCS3 expression. Vector based small hairpin RNA (shRNA) was self-designed, multiplied by E. coli and transfected in choriocarcinoma cells. Transfected cells were selected by neomycin resistance. After stimulation with Leukemia Inhibitory Factor (LIF), STAT3 phosphorylation, SOCS3 expression, proliferation and invasion was measured.
We can correlate comprehensive ultrasonographic data with various and distinct mechanisms that were described in animal models as being involved in implantation failure. Keywords: Cytokines; Implantation; Implantation; Vascularisation doi:10.1016/j.jri.2006.08.019 [O4] Interleukin 1 beta is induced by interleukin 11 during decidualization of human endometrial stromal cells, but is not released in a bioactive form C.A. White a,b, , E. Dimitriadis a , A.M. Sharkey c , C. Stoikos a , L. Salamonsen a
147
release. Bioactive IL1B was detected in vivo at very low levels and at discrete foci in late secretory phase and first trimester decidua. This regulation of latent and bioactive IL1B at the fetal–maternal interface may prime decidual cells to respond rapidly to immunological challenge or to signals from the blastocyst during implantation.
Reference White, C.A., Dimitriadis, E., Sharkey, A.M., Salamonsen, L.A., 2005. Interleukin-11 inhibits expression of insulin-like growth factor binding protein-5 mRNA in decidualizing human endometrial stromal cells. Mol Hum Reprod 11, 649–658.
Keywords: Cytokines; Chemokines and signalling; Decidua; Implantation
a
Prince Henry’s Institute of Medical Research, Australia; b Monash University, Australia; c University of Cambridge, UK Blastocyst implantation is dependent on the differentiation of endometrial stromal cells (ESC) into decidual cells. Decidualization of human ESC in vitro is enhanced by interleukin 11 (IL11), with associated changes in gene expression. Our previous microarray study identified interleukin 1 beta (IL1B) as the most upregulated gene during IL11-enhanced decidualization [1]. This study aimed to examine the effect of IL11 on interleukin 1 beta (IL1B) mRNA and protein expression during in vitro decidualization, and determine the functional significance of increased IL1B levels during this process. Stromal cells were isolated from endometrial biopsies (n = 21) obtained between days 7 and 26 of the menstrual cycle from women with no known endometrial dysfunction. Cells were decidualized with 17-estradiol and medroxyprogesterone acetate in the presence or absence of exogenous IL11, and IL1B mRNA was quantified by real-time RT–PCR. Inactive proIL1B and bioactive IL1B in cell lysates and conditioned media were measured using specific immunoassays. Secretion of bioactive IL1B from decidualizing ESC was investigated by in vitro stimulation with lipopolysaccharide, interferon gamma or human chorionic gonadotropin. Immunostaining was carried out on in vitro decidualized ESC and cycling and pregnant decidua using antibodies specific for total and bioactive IL1B. Consistent with the microarray analysis, exogenous IL11 increased the abundance of IL1B mRNA in decidualizing ESC by 28-fold (range 1.3–107.1-fold), and total immunoreactive IL1B was also increased. However, this was not reflected in bioactive IL1B secretion from these cells, and none of the tested stimuli were able to induce its
doi:10.1016/j.jri.2006.08.020 [O5] Role of signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signalling 3 (SOCS3) in regulation of trophoblast invasion T.G. Poehlmann a, , S. Bachmann b , I. Rudloff a , J. R¨odiger a , C. Neblung a , E. Schleussner a a
Friedrich-Schiller-Universit¨at Jena, Germany; f¨ur Herz- und Lungenforschung, Germany b Max-Planck-Institut
Invasion of trophoblast cells requires a fine tuning, which is fundamental for correct placentation. This tuning is regulated on intracellular level. Several signal transducers and their suppressors, known from former studies and tumor invasion are expected to be involved. The activation of the Signal Transducer and Activator of Transcription (STAT)-3 correlates with the invasiveness of carcinoma and trophoblast cells. Its negative regulator, Suppressor of Cytokine Signalling (SOCS)-3 seems to be highly expressed in non invasive trophoblasts from sheep placentomae. Aim of this investigation was to Knock Down STAT3 and SOCS3 by siRNA and shRNA in human trophoblast and carcinoma cells in order to verify the role of these molecules during human placentation. siRNA was applied to choriocarcinoma cells to knock down STAT3 and SOCS3 expression. Vector based small hairpin RNA (shRNA) was self-designed, multiplied by E. coli and transfected in choriocarcinoma cells. Transfected cells were selected by neomycin resistance. After stimulation with Leukemia Inhibitory Factor (LIF), STAT3 phosphorylation, SOCS3 expression, proliferation and invasion was measured.