Pathological mechanisms of antiphospholipid antibodies in trophoblastic cell fusion

s / Placenta 35 (2014) A1eA112 A58 Objectives: Phagocytosis is crucial for tissue homeostasis and remodelling in addition to its role in defending against invading organisms and allogeneic cells. A “Don’t-eat-me” signal generated by binding of signal regulatory protein alpha (SIRPa) on the surface of macrophages and its ubiquitously expressed ligand, CD47, plays an important role in regulating phagocytic activity. Our previous results show that pro-inflammatory stimulus-activated first trimester decidual cells (FTDCs) enhance the activity of macrophages in inducing apoptosis in first trimester extravillous trophoblasts (EVTs). Seeking to enhance understanding of decidual remodeling during pregnancy, this proposal aims to study themechanisms of FTDC/macrophage interactions resulting in previously undisclosed homeostasis on placentation. Methods: FTDCs were primed with estradiol (E) (10 8 M) + medroxyprogesterone acetate (M) (10 7 M) for 7d followed by incubating with 1 ng/ml of interleukin-1 beta (IL-1b) or tumor necrosis factor-alpha (TNF-a) for 24h. Peripheral monocyte-derived macrophages were incubated with conditioned media (CM) from these FTDC cultures for 2d. The phagocytic activity of peripheral monocyte-derived macrophages was examined by co-culturing with apoptotic bodies derived from PKH67-labeled HTR-8/ SVneo (HTR-8) cells, a first trimester EVT cell line. The phagocytosis of HTR apoptotic bodies by Mfs was evaluated by fluorescent microscopy and flow cytometric analysis. SIRPa expression on macrophages was examined by flow cytometric analysis, qRT-PCR and Western blot analysis. Results: CM from IL-1bor TNF-a-treated FTDCs consistently enhanced phagocytosis of HTR-8-derived apoptotic bodies by macrophages, while SIRPa expression was suppressed. Conclusions: These results suggest that FTDCs may play a role in maintaining decidual homeostasis by promoting phagocytic activity of macrophages upon remodelling in response to pro-inflammatory stimuli. However, the enhanced phagocytic activity can also lead to increased killing of invading semi-allogeneic EVTs. P1.152. PREGNANCY RECALL IN DECIDUAL NK CELLS Debra Goldman-Wohl , Moriya Gamliel , Ronit Gilad , Tal Imbar , Ronit Haimov-Kochman , Caryn Greenfield , Iris Eisenberg-Loebl , Ofer Mandelboim , Simcha Yagel a a The Magda and Richard Hoffman Center for Human Placenta Research, Department of Obstetrics and Gynecology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel; b Lautenberg Center for Immunology, Hebrew University Hadassah Medical School, Jerusalem, Israel Objective: Natural killer (NK) cells, lymphocytes of the innate immune system, act to kill tumor and infected cells. However, recent studies suggest that NK cells possess some features of adaptive immunity including immune memory. Specifically, the expansion of the NKG2C+ NK cell population is implicated in the “memory” response of NK cells to HCMV infection. NK cells of the decidua (dNK) possess unique phenotypical and functional properties and are considered regulators of remodeling of the maternal fetal interface. Here we investigate if these dNK cells have the ability to “remember” pregnancy. Methods: dNK cells from nulliparous and parous women were isolated from first trimester elective pregnancy terminations .The dNK cell receptor repertoire was analyzed by FACS. Transcriptome analysis was performed using RNA-seq. Results: Significantly more NKG2C+ dNK cells (P1⁄40.0008 (2-tailed)) were detected in the parous versus nulliparous women. This finding was specific for the dNK population but not observed in T cells, and did not correlate with CMV serology status. In parous samples, transcriptome analysis revealed upregulation of genes related to angiogenesis, tissue remodeling, TH1 to TH2 shift, secretion of growth factors and NK cell function/ recruitment. The most significant transcriptome results were validated by real-time PCR; they include PAEP, IGFBP1 and SPP1. Conclusion: This observation suggests that dNK cells that play a role in remodeling the maternal fetal interface may additionally possess the ability to “recall” pregnancy. This findingmay give insight to the etiology of preeclampsia with increased risk in first pregnancy. P1.153-N. CYTOKINES PROLIFE IN MATERNAL AND CORD BLOOD OF DIABETIC MOTHERS Glilciane Morceli , Cristiane Hara , Adenilda Hon orio-Franca , Eduardo Franca , Danny Laura Fagundes , Marilza Rudge , D ebora Damasceno , Iracema Calderon a Botucatu Medical School, Botucatu/S~ ao Paulo, Brazil; b Institute of Biological and Health Science Federal University of Mato Grosso, Barra do Garcas/Mato Grosso, Brazil Background: Diabetes is an inflammatory condition and cytokines have been implicated in its physiopathology. Thus, the interaction between cytokines pro-inflammatory and anti-inflammatory is essential for pregnancy outcome. Objective: Evaluated the cytokine profile in maternal and cord blood from pregnancies complicated by hyperglycemia. Methods: The women were classified into three groups, according to their glycemic status: normoglycemic (N 1⁄4 15), mild hyperglycemic (N 1⁄4 15) and diabetic (N 1⁄4 15). The cytokines levels were determined by flow cytometry and the data were analyses by software FCAP Array 1.0. Results: No significant changes were detected in the IL2, IL4 and IFN-g concentrations between maternal and cord blood, neither among three groups. IL6 and IL10 concentrations were increased in cord blood, and IL17 levels were higher in maternal blood of diabetic mothers. Maternal blood IL10 and IL17 concentrations were found significantly correlated with levels observed in the umbilical cord. Conclusions: These data suggest that diabetes modifies the cytokines profile in maternal and cord blood. These results corroborate the hypothesis that diabetes alters the role of cytokines, and thus may influence the systemic and mucosal immune response.