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Trophoblast migration is activated via chemokine receptors 1 and 3
Posters / Journal of Reproductive Immunology 90 (2011) 164–183
P6 Trophoblast migration is activated via chemokine receptors 1 and 3 I. Knöfler a,b,∗ , C. Röhler a,b , S. Hölters a , J.S. Fitzgerald a , D.M. Morales Prieto a , M. Wartenberg b , E. Schleussner a , U.R. Markert a a
Placenta-Labor, Department of Obstetrics, Germany Clinic of Internal Medicine I, Cardiology Division, University Hospital Jena, 07740 Jena, Germany b
Introduction: Cytotrophoblast cells invade the decidua, manly towards the myometrium and maternal blood vessels. The responsible (chemo-) attractants are not completely identified. Methods: We used the immortalized first trimester trophoblast cell line HTR8/svneo to form spheroids with a diameter of approximately 700 m and confronted them to human placental tissue explants of similar size from different regions. Before confrontation, spheroid cells were stained with fluorescent green Mito Tracker, blood vessels were stained after confrontation red with anti-CD31 antibodies. Finally, a part of spheroids were incubated with the specific blocker chemokine receptor 1 (CCR1) and CCR3 UCB 35625. A total of 200 different confrontation products were analyzed after 10, 24 and 48 h by using a laser scanning microscope. Results: After 10 h, HTR8/svneo cells start to leave spheroids, which after 48 h are completely disaggregated. Cells invade decidual tissue and cover villi. These effects are completely inhibited when blocked CCR1/3 is blocked. Conclusion: Approximately 10 different possible CCR1/3 candidate ligands are known to be present in human placenta. At least one chemokine which binds to CCR1/3 is responsible for directed trophoblast migration and invasion in the decidua. doi:10.1016/j.jri.2011.06.065 P7 The immunological aspects of infertile women who resort to IVF-ET Y. Zeng a,b,∗ , P. Liang a,b , M. Mo a,b , L. Wang a,b , G.-G. Li a,b , B. Yin a,b , J. Cai a,b , T. Wu a,b
167
and the day of HCG administration following the COH. After the IVF-ET, the infertile women were assigned to two groups according to the clinical pregnancy result: a pregnant group (14 women) and a non-pregnant group (19 women). One woman was excluded because of ectopic pregnancy. The parameters measured were as follows: percentages and absolute counts of T cells, B cells and NK cells as well as the CD4+ and CD8+ subpopulations of T cells in peripheral blood; NK cytotoxicity; Th1: Th2 Cytokine assay, the percentages of CD19+ CD5+ B cells, HLA-DR+ T cells, CD69+ T cells, CD69+ NK cells, CD56dim CD16+ NK cells, CD56bright CD16− NK cells, and CD4+ CD25+ CD127− Treg cells. Results: The IFN␥/IL-10 and TNF␣/IL-10 ratios of the non-pregnant group were significantly higher compared with the pregnant group on the day of HCG administration. There was no significant difference either on the beginning day of GnRHa administration or the beginning day of controlled ovarian hyperstimulation (COH) in the two groups. In the non-pregnant group, the TNF␣ and IFN␥ decreased significantly after the GnRHa administration, the absolute counts of CD3+ , CD3+ CD8+ , CD56+ CD16+ , CD19+ cell decreased respectively both after the GnRHa administration and controlled ovarian hyperstimulation; while the CD69+ T cell and the CD69+ NK cells increased after the controlled ovarian hyperstimulation. In the pregnant group, the CD3+ CD4+ HLA-DR+ and CD69+ NK cells increased after the controlled ovarian hyperstimulation, while the Treg cells decreased both after the GnRHa administration and controlled ovarian hyperstimulation. Conclusions: The immunological aspects especially the Th1: Th2 cytokine of the infertile woman may play an important role in reproductive immunology and the success or failure of embryo implantation in the process of IVF-ET. Keywords: IVF-ET; Lymphocyte; NK cytotoxicity; Th1: Th2 cytokine doi:10.1016/j.jri.2011.06.066 P8 Effects of STAT1 suppression on ERK1/2 in trophoblastic cells F.L. Pereira de Sousa ∗ , D.M. Morales Prieto, S. Ospina, W. Chaiwangyen, S. Daher, N. Sass, U.R. Markert
a
Clinical Center for Recurrent Implantation Failure & Recurrent Spontaneous Abortion, Fertility Center, Shenzhen Zhongshan Urological Hospital, Shenzhen, China b Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics, Shenzhen, China
Introduction: To study the immunological aspects of infertile women who resort to IVF-ET. Methods: In a retrospective randomized double-blind comparison study, 34 infertile women with normal ovarian reserve were selected, and all received a controlled ovarian hyperstimulation (COH) protocol using the conventional GnRHa long protocol. The immunological aspects were evaluated at three time points: the beginning day of GnRHa administration, the beginning day of COH,
Placenta-Laboratories, Department of Obstetrics, University Hospital Jena, Germany Introduction: Migration and trophoblast invasion are controlled functionally along with the active participation of cytokines and growth factors. Two important intracellular signaling pathways are the Janus kinase/signal transducer and activator of transcription (JAK–STAT) and extracellular regulated kinase1/2 (ERK1/2). These pathways have been associated with the regulation of gene expression, cellular proliferation, differentiation, angiogenesis, embryo development and invasion in tumor and trophoblast cells. The aim of our study is to characterize and analyze the regulation and crosstalks of STAT1 and
P6 Trophoblast migration is activated via chemokine receptors 1 and 3 I. Knöfler a,b,∗ , C. Röhler a,b , S. Hölters a , J.S. Fitzgerald a , D.M. Morales Prieto a , M. Wartenberg b , E. Schleussner a , U.R. Markert a a
Placenta-Labor, Department of Obstetrics, Germany Clinic of Internal Medicine I, Cardiology Division, University Hospital Jena, 07740 Jena, Germany b
Introduction: Cytotrophoblast cells invade the decidua, manly towards the myometrium and maternal blood vessels. The responsible (chemo-) attractants are not completely identified. Methods: We used the immortalized first trimester trophoblast cell line HTR8/svneo to form spheroids with a diameter of approximately 700 m and confronted them to human placental tissue explants of similar size from different regions. Before confrontation, spheroid cells were stained with fluorescent green Mito Tracker, blood vessels were stained after confrontation red with anti-CD31 antibodies. Finally, a part of spheroids were incubated with the specific blocker chemokine receptor 1 (CCR1) and CCR3 UCB 35625. A total of 200 different confrontation products were analyzed after 10, 24 and 48 h by using a laser scanning microscope. Results: After 10 h, HTR8/svneo cells start to leave spheroids, which after 48 h are completely disaggregated. Cells invade decidual tissue and cover villi. These effects are completely inhibited when blocked CCR1/3 is blocked. Conclusion: Approximately 10 different possible CCR1/3 candidate ligands are known to be present in human placenta. At least one chemokine which binds to CCR1/3 is responsible for directed trophoblast migration and invasion in the decidua. doi:10.1016/j.jri.2011.06.065 P7 The immunological aspects of infertile women who resort to IVF-ET Y. Zeng a,b,∗ , P. Liang a,b , M. Mo a,b , L. Wang a,b , G.-G. Li a,b , B. Yin a,b , J. Cai a,b , T. Wu a,b
167
and the day of HCG administration following the COH. After the IVF-ET, the infertile women were assigned to two groups according to the clinical pregnancy result: a pregnant group (14 women) and a non-pregnant group (19 women). One woman was excluded because of ectopic pregnancy. The parameters measured were as follows: percentages and absolute counts of T cells, B cells and NK cells as well as the CD4+ and CD8+ subpopulations of T cells in peripheral blood; NK cytotoxicity; Th1: Th2 Cytokine assay, the percentages of CD19+ CD5+ B cells, HLA-DR+ T cells, CD69+ T cells, CD69+ NK cells, CD56dim CD16+ NK cells, CD56bright CD16− NK cells, and CD4+ CD25+ CD127− Treg cells. Results: The IFN␥/IL-10 and TNF␣/IL-10 ratios of the non-pregnant group were significantly higher compared with the pregnant group on the day of HCG administration. There was no significant difference either on the beginning day of GnRHa administration or the beginning day of controlled ovarian hyperstimulation (COH) in the two groups. In the non-pregnant group, the TNF␣ and IFN␥ decreased significantly after the GnRHa administration, the absolute counts of CD3+ , CD3+ CD8+ , CD56+ CD16+ , CD19+ cell decreased respectively both after the GnRHa administration and controlled ovarian hyperstimulation; while the CD69+ T cell and the CD69+ NK cells increased after the controlled ovarian hyperstimulation. In the pregnant group, the CD3+ CD4+ HLA-DR+ and CD69+ NK cells increased after the controlled ovarian hyperstimulation, while the Treg cells decreased both after the GnRHa administration and controlled ovarian hyperstimulation. Conclusions: The immunological aspects especially the Th1: Th2 cytokine of the infertile woman may play an important role in reproductive immunology and the success or failure of embryo implantation in the process of IVF-ET. Keywords: IVF-ET; Lymphocyte; NK cytotoxicity; Th1: Th2 cytokine doi:10.1016/j.jri.2011.06.066 P8 Effects of STAT1 suppression on ERK1/2 in trophoblastic cells F.L. Pereira de Sousa ∗ , D.M. Morales Prieto, S. Ospina, W. Chaiwangyen, S. Daher, N. Sass, U.R. Markert
a
Clinical Center for Recurrent Implantation Failure & Recurrent Spontaneous Abortion, Fertility Center, Shenzhen Zhongshan Urological Hospital, Shenzhen, China b Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics, Shenzhen, China
Introduction: To study the immunological aspects of infertile women who resort to IVF-ET. Methods: In a retrospective randomized double-blind comparison study, 34 infertile women with normal ovarian reserve were selected, and all received a controlled ovarian hyperstimulation (COH) protocol using the conventional GnRHa long protocol. The immunological aspects were evaluated at three time points: the beginning day of GnRHa administration, the beginning day of COH,
Placenta-Laboratories, Department of Obstetrics, University Hospital Jena, Germany Introduction: Migration and trophoblast invasion are controlled functionally along with the active participation of cytokines and growth factors. Two important intracellular signaling pathways are the Janus kinase/signal transducer and activator of transcription (JAK–STAT) and extracellular regulated kinase1/2 (ERK1/2). These pathways have been associated with the regulation of gene expression, cellular proliferation, differentiation, angiogenesis, embryo development and invasion in tumor and trophoblast cells. The aim of our study is to characterize and analyze the regulation and crosstalks of STAT1 and