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Abstracts / Placenta 35 (2014) A1eA112 A57

B) ProtonMagnetic resonance spectroscopy was performed to evaluate thebiochemical/ metabolic profile in placentas. A significant decrease inphosphocholine was observed in APS-mice compared to Ctrl (PCh/Cr+PCr:APS(n¼7)¼0.50±0.78 vs control (n¼4)¼1.69±0.73, p¼0.0352). Decreasedglucose levels and increased lactate levels, indicative of hypoxia, weremeasured in placentas from APS-mice compared to Ctrl (Glc/Cr+PCr:APS(n¼6)¼ 0.03±0.0.05 vs control (n¼4)¼1.04±0.9, p¼0.0222; Lac/Cr+PCr:APS(n¼7): 0.47±0.0.80 vs control (n¼4): 4.4±4.1, p¼0.0310). Placentalinsufficiency in APS was also associated with abnormal fetal neuro-development characterised by cortical axonal cytoarchitecture dis-ruption(NF200 and MAP-2 expression) and increased neurodegeneration(FluoroJadeB). Increased anxietywas observed in the offspring of APS-mice.

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Conclusion: Using two novel non invasive in uterus MRI-based methodswe were able to determine complement activation and metabolism in theplacentas and predict fetal outcomes.

P1.149-N.PATHOLOGICAL MECHANISMS OF ANTIPHOSPHOLIPID ANTIBODIES INTROPHOBLASTIC CELL FUSION

Tess Marchetti a,b, Philippe de Moerloose b, Marie Cohen a a Laboratory ofHormonology, Geneva University Hospitals, Geneva, Switzerland;bHaemostasis Unit, Geneva University Hospitals, Geneva, Switzerland

Objectives: Obstetrical Antiphospholipid Syndrome (APS) associatespregnancy pathologies with antiphospholipid antibodies (aPL). It has beensuggested that aPL could affect trophoblastic cell fusion. However, theirpathogenicity is still poorly understood.

Toll-like Receptors (TLR) have been implicated in the pathological activa-tion of aPL on endothelial cells. Although inflammation has been incrim-inated in this process, endoplasmic reticulum (ER) stress activation seemsmore related to trophoblastic fusion impairment by aPL.The aim of our study was to evaluate the pathological mechanisms of aPLon trophoblastic cell fusion.Methods:BeWocell line is usedas amodel tomimic trophoblastic cell fusion,triggered by forskolin. Results were corroborated with primary cytotropho-blastic cells. Fusion index was determined by immunocytochemistry andbiochemical differentiation by ELISA measuring hCG secretion.Effects of TLR on BeWo cell fusionwere evaluated using blocker antibodiesfor TLR2 and 4, peptide binding for TLR2 and 4, and stable shRNA TLR4 inthe presence or not of aPL.

After determining the role of TLR4 in trophoblastic cell fusion impairmentby aPL, signalling cascade of inflammationwas studied by ELISAmeasuringTNFa and IL8 secretion. ER stress activation was assessed by qPCR and WBquantifying Glucose-regulated protein 78 (GRP78) and C/EBP-homologousprotein (CHOP) expressions.Result: Fusion index and hCG secretion are decreased by addition of aPL. Inthe presence of aPL and anti-TLR4 antibodies or in BeWo cells transfectedwith shRNA TLR4, trophoblastic cell fusion and hCG secretion wererestored.Neither TNFa nor IL8 secretion was observed in BeWo cells stimulated byaPL. Increase in GRP78 and CHOP expressions was observed in cellsstimulated by aPL, but not when blocked with anti-TLR4 antibodies, sug-gesting a role of ER stress in trophoblastic cell fusion impairment by aPL.Conclusion: aPL interfered with fusion process and biochemical differ-entiation of trophoblastic cells via TLR4 that triggers ER stress activation.

P1.150.EXAMINING THE EFFECT OF OBESITY-ASSOCIATED SYSTEMICINFLAMMATION ON THE UTERINE IMMUNE CELL NICHE IN EARLYPREGNANCY

Yoona Kim, Sofie Perdu, Mahroo Aghababaei, Alexander Beristain TheUniversity of British Columbia, Vancouver, Canada

Objectives: Low-grade inflammation associates with major obstetricaldisorders like preterm birth and preeclampsia and is thought to be causalin the development of these high-risk pregnancies. Control of immunecell-directed pro-inflammatory cytokine production and cytotoxicity iscrucial for utero-placental function; aberrant pro-inflammatory T helper(h) 1 cell activity within the uterine microenvironment associates withplacental dysfunction and poor pregnancy outcome. Immunological im-balances and low-grade inflammation are associatedwith excess adiposity.However, the cellular mechanism(s) linking obesity-associated inflam-mation to poor pregnancy outcome are poorly understood. As a first step inaddressing this knowledge gap, our study aims to examine multiple im-mune cell subsets within decidual tissues of non-obese and obese womenin early pregnancy.

Methods: Patient information, blood serum and decidual tissue fromgestationally aged-matched (8-10 weeks) women undergoing electivepregnancy terminations were used to determine BMI, serum C-reactiveprotein (CRP) (Qg/mL) and immune cell composition. Women were cate-gorized as healthy BMI (20-24.9) not presenting with low-grade inflam-mation (CRP < 2 Qg/mL; ControlCRP-) or obese BMI (� 30) presenting withlow-grade inflammation (CRP � 4 Qg/mL; ObeseCRP+). Immune cells,including natural killer cells (NKs), T cells (Th1, Th2, Th17, Tregs and CD8+

cytotoxic cells), macrophages and dendritic cells, were quantitated bymulticolour flow cytometry analysis.Results:Wedid not detect differences in proportions of CD56bright decidualNKs, dendritic cells, macrophages or Th17 and CD8+ T cells betweendecidual tissues of ControlCRP- (n¼15) and ObeseCRP+ (n¼15) women.However, proportions of immunosuppressive CD4+/FoxP3+ Tregs, pro-in-flammatory IFN©+ Th1 cells, and cytotoxic CD16+ NK cells were elevated inObeseCRP+ decidua while IL-13+ Th2 proportions were decreased.Conclusion: This study suggests that obesity-linked inflammation inearly pregnancy does alter immune cell composition within thematernal-fetal interface. These new findings lay the groundwork forfuture studies aimed at dissecting the importance of these cellularchanges in pregnancy.

P1.151.PHAGOCYTIC ACTIVITY OF MACROPHAGES IS REGULATED BY FIRSTTRIMESTER DECIDUAL CELLS IN RESPONSE TO PRO-INFLAMMATORYSTIMULI VIA THE MODULATION OF “DON'T-EAT-ME” SIGNAL

Longzhu Piao a, Susheela Tridandapani b, S. Joseph Huang a aDeartment ofOb/Gyn, The Ohio State University, Columbus, OH, USA; b SBS-MicrobialInfection/Immunity, The Ohio State University, Columbus, OH, USA