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Explore the true uterine NK cell cytokines production by the stimulation using semen
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Abstracts / Journal of Reproductive Immunology 115 (2016) 71–90
DHA. Concentration of IL-1 and IL-18, and activation of caspase-1 in supernatants were measured by ELISA. Results: Both EPA and DHA significantly reduced NLRP3induced IL-1 (EPA-ATP 41.0 ± 10.7%, EPA-Nig 30.5 ± 5.7%, DHAATP 29.4% ± 9.8%, DHA-Nig 27.7 ± 10.3%) and IL-18 (EPA-ATP 66.7% ± 10.7%, EPA-Nig 36.1% ± 12.2%, DHA-ATP 58.7% ± 17.0%, DHA-Nig 41.0 ± 16.4%) secretions, and caspase-1 activation (EPAATP 93.5% ± 6.7%, EPA-Nig 73.9 ± 7.0%, DHA-ATP 87.8% ± 5.1%, DHA-Nig 87.1 ± 12.7%) in DM. Conclusions: -3FAs reduced NLRP3-induced inflammation in DM, which may explain the positive impacts of -3 FA on the prevention of preeclampsia. http://dx.doi.org/10.1016/j.jri.2016.04.235 P2-17 Endometrial gene expression of women with recurrent pregnancy losses and infertility Li Wu 1 , Nayoung Sung 1 , Maria Dinorah Salazar Garcia 1 , Annie Skariah 1 , Svetlana Dambaeva 2 , Kenneth Beaman 2 , Alice Gilman-Sachs 2 , Joanne Kwak-Kim 1,2 1 Reproductive Medicine, Department of Obstetrics and Gynecology, USA 2 Department of Microbiology and Immunology at Chicago Medical School at Rosalind Franklin University of Medicine and Science, Chicago, OH, USA
Problem: While the relationship between the various gene expression and immune effectors in endometrium remains unknown, the expression of these genes could be utilized as a biomarker for of optimal uterine receptivity of women with a history of recurrent pregnancy losses (RPL) and infertility. Material and methods: A prospective controlled study was carried out in women with RPL or infertility. Endometrial biopsy is performed during luteal phase and endometrium gene expression was investigated with quantify PCR for IL-18, TWEAK, IL-15, Fn14, TGF-, NF- B, CSF-R and ROR ␥ T. In additional, peripheral blood NK cell cytotoxicity and Th1/Th2 cell ratios were analyzed by flow cytometry. Results: Patients with RPL had a significantly higher gene expression of IL-18 and the ratio of IL-18/TWEAK when compared with those of infertility group (P < 0.05, respectively). The ratio of IL15/FN-14 in PRL group was lower than those of infertility group. NK cytotoxicity, the ratios of IFN-␥/IL-10 and TNF-␣/IL-10 were higher than controls, but there are no differences between the two groups. TGF-, NF-B, CSF-R and ROR␥ T gene expressions were found in both groups but there were no difference between the two groups. Conclusion: Endometrial IL-18 and the ratio of IL-18/TWEAK gene expression are up-regulated in patient with RPL. Further study is warranted to investigate if these gene expressions are correlated with uNK cells activation and predict pregnancy outcome. http://dx.doi.org/10.1016/j.jri.2016.04.236
P2-18 Explore the true uterine NK cell cytokines production by the stimulation using semen Ayako Taima, Atsushi Fukui, Kohei Fuchinoue, Hitomi Chiba, Mai Kamoi, Ayano Funamizu, Hideki Mizunuma Department of Obstetrics and Gynecology, Graduate School of Medicine, Hirosaki University, Hirosaki, Aomori, Japan Objectives: Some of women with recurrent pregnancy loss have an abnormality of NK cell cytokines production. To evaluate cytokines production from immune cells such as NK cell, T cell and so on, PMA and Ionomycin are used as a stimulator of cells. PMA activates protein tyrosine kinase and Ionomycin increases Ca2+ levels. Exposure of semen is essential at the time of pregnancy and semen seems to be an immunomodulator. We aimed to develop the physiological stimulation method of immune cells for reproductive immunology field. Methods: Uterine endometrial samples were obtained from women with infertility (n = 26). To evaluate the cytokines production by uterine NK (uNK) cells, uNK cells were stimulated by PMA/Ionomycin (n = 19) or semen (n = 70). Cytokines (IFN-␥, TNF-␣, IL-4, IL-10) producing NK cell was evaluated by flow cytometry. Results: The percentage of IFN-␥ or TNF-␣ producing NK cell stimulated by semen was significantly higher than without stimulation (IFN-␥: p < 0.05, TNF-␣: p < 0.01) and lower than stimulated by PMA/Ionomycin (IFN-␥: p < 0.01, TNF-␣: p < 0.01). The percentage of IL-4 of IL-10 producing NK cell stimulated by semen was the highest. The percentage of IFN-␥ producing NK cell stimulated by semen was not different from that stimulated by seminal plasma but significantly higher than by sperm (p < 0.05). Conclusions: We firstly reported that it was possible to evaluate the cytokines production by uNK cells stimulated by semen. http://dx.doi.org/10.1016/j.jri.2016.04.237 P2-19 Expression and functional analysis of C19MC microRNAs in normal and preeclamptic pregnancies Daniel Fabig, Ivan Dimitrov, Udo R. Markert, Diana M. Morales-Prieto University Hospital Jena, Department of Obstetrics, Placenta Lab, Jena, Germany Background: Trophoblast cells have a specific miRNA profile that includes the chromosome 19 microRNA cluster (C19MC). Dysregulation of C19MC miRNAs may be associated with development of pregnancy pathologies. Here, expression and function of C19MC members and their association with preeclampsia (PE) have been studied in placenta tissue and trophoblastic cells. Methods: Total RNA was isolated from placenta tissue of normal and PE patients. PE group was divided into early (≤34th) or late (>34th week) delivery. Expression of ten microRNAs from C19MC miRNAs was analyzed by individual qPCR. Trophoblastic cells were transfected with either mimics or inhibitors of miR-520d-3p or the respective controls. Effects on JEG-3 and HTR-8/SVneo cell proliferation and invasion were investigated by MTS and Matrigel assays. Results: Four miRNAs (miR-520d-3p, -525-3p, -519d and -518b) were upregulated (>1.5-fold) in the early delivery PE group compared to normal pregnancies. Preliminary results demonstrate that upon miR-520d-3p inhibition, in JEG-3 cells invasion increased
Abstracts / Journal of Reproductive Immunology 115 (2016) 71–90
DHA. Concentration of IL-1 and IL-18, and activation of caspase-1 in supernatants were measured by ELISA. Results: Both EPA and DHA significantly reduced NLRP3induced IL-1 (EPA-ATP 41.0 ± 10.7%, EPA-Nig 30.5 ± 5.7%, DHAATP 29.4% ± 9.8%, DHA-Nig 27.7 ± 10.3%) and IL-18 (EPA-ATP 66.7% ± 10.7%, EPA-Nig 36.1% ± 12.2%, DHA-ATP 58.7% ± 17.0%, DHA-Nig 41.0 ± 16.4%) secretions, and caspase-1 activation (EPAATP 93.5% ± 6.7%, EPA-Nig 73.9 ± 7.0%, DHA-ATP 87.8% ± 5.1%, DHA-Nig 87.1 ± 12.7%) in DM. Conclusions: -3FAs reduced NLRP3-induced inflammation in DM, which may explain the positive impacts of -3 FA on the prevention of preeclampsia. http://dx.doi.org/10.1016/j.jri.2016.04.235 P2-17 Endometrial gene expression of women with recurrent pregnancy losses and infertility Li Wu 1 , Nayoung Sung 1 , Maria Dinorah Salazar Garcia 1 , Annie Skariah 1 , Svetlana Dambaeva 2 , Kenneth Beaman 2 , Alice Gilman-Sachs 2 , Joanne Kwak-Kim 1,2 1 Reproductive Medicine, Department of Obstetrics and Gynecology, USA 2 Department of Microbiology and Immunology at Chicago Medical School at Rosalind Franklin University of Medicine and Science, Chicago, OH, USA
Problem: While the relationship between the various gene expression and immune effectors in endometrium remains unknown, the expression of these genes could be utilized as a biomarker for of optimal uterine receptivity of women with a history of recurrent pregnancy losses (RPL) and infertility. Material and methods: A prospective controlled study was carried out in women with RPL or infertility. Endometrial biopsy is performed during luteal phase and endometrium gene expression was investigated with quantify PCR for IL-18, TWEAK, IL-15, Fn14, TGF-, NF- B, CSF-R and ROR ␥ T. In additional, peripheral blood NK cell cytotoxicity and Th1/Th2 cell ratios were analyzed by flow cytometry. Results: Patients with RPL had a significantly higher gene expression of IL-18 and the ratio of IL-18/TWEAK when compared with those of infertility group (P < 0.05, respectively). The ratio of IL15/FN-14 in PRL group was lower than those of infertility group. NK cytotoxicity, the ratios of IFN-␥/IL-10 and TNF-␣/IL-10 were higher than controls, but there are no differences between the two groups. TGF-, NF-B, CSF-R and ROR␥ T gene expressions were found in both groups but there were no difference between the two groups. Conclusion: Endometrial IL-18 and the ratio of IL-18/TWEAK gene expression are up-regulated in patient with RPL. Further study is warranted to investigate if these gene expressions are correlated with uNK cells activation and predict pregnancy outcome. http://dx.doi.org/10.1016/j.jri.2016.04.236
P2-18 Explore the true uterine NK cell cytokines production by the stimulation using semen Ayako Taima, Atsushi Fukui, Kohei Fuchinoue, Hitomi Chiba, Mai Kamoi, Ayano Funamizu, Hideki Mizunuma Department of Obstetrics and Gynecology, Graduate School of Medicine, Hirosaki University, Hirosaki, Aomori, Japan Objectives: Some of women with recurrent pregnancy loss have an abnormality of NK cell cytokines production. To evaluate cytokines production from immune cells such as NK cell, T cell and so on, PMA and Ionomycin are used as a stimulator of cells. PMA activates protein tyrosine kinase and Ionomycin increases Ca2+ levels. Exposure of semen is essential at the time of pregnancy and semen seems to be an immunomodulator. We aimed to develop the physiological stimulation method of immune cells for reproductive immunology field. Methods: Uterine endometrial samples were obtained from women with infertility (n = 26). To evaluate the cytokines production by uterine NK (uNK) cells, uNK cells were stimulated by PMA/Ionomycin (n = 19) or semen (n = 70). Cytokines (IFN-␥, TNF-␣, IL-4, IL-10) producing NK cell was evaluated by flow cytometry. Results: The percentage of IFN-␥ or TNF-␣ producing NK cell stimulated by semen was significantly higher than without stimulation (IFN-␥: p < 0.05, TNF-␣: p < 0.01) and lower than stimulated by PMA/Ionomycin (IFN-␥: p < 0.01, TNF-␣: p < 0.01). The percentage of IL-4 of IL-10 producing NK cell stimulated by semen was the highest. The percentage of IFN-␥ producing NK cell stimulated by semen was not different from that stimulated by seminal plasma but significantly higher than by sperm (p < 0.05). Conclusions: We firstly reported that it was possible to evaluate the cytokines production by uNK cells stimulated by semen. http://dx.doi.org/10.1016/j.jri.2016.04.237 P2-19 Expression and functional analysis of C19MC microRNAs in normal and preeclamptic pregnancies Daniel Fabig, Ivan Dimitrov, Udo R. Markert, Diana M. Morales-Prieto University Hospital Jena, Department of Obstetrics, Placenta Lab, Jena, Germany Background: Trophoblast cells have a specific miRNA profile that includes the chromosome 19 microRNA cluster (C19MC). Dysregulation of C19MC miRNAs may be associated with development of pregnancy pathologies. Here, expression and function of C19MC members and their association with preeclampsia (PE) have been studied in placenta tissue and trophoblastic cells. Methods: Total RNA was isolated from placenta tissue of normal and PE patients. PE group was divided into early (≤34th) or late (>34th week) delivery. Expression of ten microRNAs from C19MC miRNAs was analyzed by individual qPCR. Trophoblastic cells were transfected with either mimics or inhibitors of miR-520d-3p or the respective controls. Effects on JEG-3 and HTR-8/SVneo cell proliferation and invasion were investigated by MTS and Matrigel assays. Results: Four miRNAs (miR-520d-3p, -525-3p, -519d and -518b) were upregulated (>1.5-fold) in the early delivery PE group compared to normal pregnancies. Preliminary results demonstrate that upon miR-520d-3p inhibition, in JEG-3 cells invasion increased