- Email: [email protected]
Phagocytic activity of macrophages is regulated by first trimester decidual cells in response to pro-inflammatory stimuli via the modulation of “Don't-Eat-Me” signal
Abstracts / Placenta 35 (2014) A1eA112
Ă
B) Proton Magnetic resonance spectroscopy was performed to evaluate the biochemical/ metabolic profile in placentas. A significant decrease in phosphocholine 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). Decreased glucose levels and increased lactate levels, indicative of hypoxia, were measured 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). Placental insufficiency in APS was also associated with abnormal fetal neurodevelopment characterised by cortical axonal cytoarchitecture disruption(NF200 and MAP-2 expression) and increased neurodegeneration (FluoroJadeB). Increased anxiety was observed in the offspring of APS-mice.
A57
After determining the role of TLR4 in trophoblastic cell fusion impairment by aPL, signalling cascade of inflammation was studied by ELISA measuring TNFa and IL8 secretion. ER stress activation was assessed by qPCR and WB quantifying Glucose-regulated protein 78 (GRP78) and C/EBP-homologous protein (CHOP) expressions. Result: Fusion index and hCG secretion are decreased by addition of aPL. In the presence of aPL and anti-TLR4 antibodies or in BeWo cells transfected with shRNA TLR4, trophoblastic cell fusion and hCG secretion were restored. Neither TNFa nor IL8 secretion was observed in BeWo cells stimulated by aPL. Increase in GRP78 and CHOP expressions was observed in cells stimulated by aPL, but not when blocked with anti-TLR4 antibodies, suggesting a role of ER stress in trophoblastic cell fusion impairment by aPL. Conclusion: aPL interfered with fusion process and biochemical differentiation of trophoblastic cells via TLR4 that triggers ER stress activation.
P1.150. EXAMINING THE EFFECT OF OBESITY-ASSOCIATED SYSTEMIC INFLAMMATION ON THE UTERINE IMMUNE CELL NICHE IN EARLY PREGNANCY Yoona Kim, Sofie Perdu, Mahroo Aghababaei, Alexander Beristain The University of British Columbia, Vancouver, Canada Objectives: Low-grade inflammation associates with major obstetrical disorders like preterm birth and preeclampsia and is thought to be causal in the development of these high-risk pregnancies. Control of immune cell-directed pro-inflammatory cytokine production and cytotoxicity is crucial for utero-placental function; aberrant pro-inflammatory T helper (h) 1 cell activity within the uterine microenvironment associates with placental dysfunction and poor pregnancy outcome. Immunological imbalances and low-grade inflammation are associated with excess adiposity. However, the cellular mechanism(s) linking obesity-associated inflammation to poor pregnancy outcome are poorly understood. As a first step in addressing this knowledge gap, our study aims to examine multiple immune cell subsets within decidual tissues of non-obese and obese women in early pregnancy.
Ă
Conclusion: Using two novel non invasive in uterus MRI-based methods we were able to determine complement activation and metabolism in the placentas and predict fetal outcomes.
P1.149-N. PATHOLOGICAL MECHANISMS OF ANTIPHOSPHOLIPID ANTIBODIES IN TROPHOBLASTIC CELL FUSION Tess Marchetti a, b, Philippe de Moerloose b, Marie Cohen a a Laboratory of Hormonology, Geneva University Hospitals, Geneva, Switzerland; b Haemostasis Unit, Geneva University Hospitals, Geneva, Switzerland Objectives: Obstetrical Antiphospholipid Syndrome (APS) associates pregnancy pathologies with antiphospholipid antibodies (aPL). It has been suggested that aPL could affect trophoblastic cell fusion. However, their pathogenicity is still poorly understood. Toll-like Receptors (TLR) have been implicated in the pathological activation of aPL on endothelial cells. Although inflammation has been incriminated in this process, endoplasmic reticulum (ER) stress activation seems more related to trophoblastic fusion impairment by aPL. The aim of our study was to evaluate the pathological mechanisms of aPL on trophoblastic cell fusion. Methods: BeWo cell line is used as a model to mimic trophoblastic cell fusion, triggered by forskolin. Results were corroborated with primary cytotrophoblastic cells. Fusion index was determined by immunocytochemistry and biochemical differentiation by ELISA measuring hCG secretion. Effects of TLR on BeWo cell fusion were evaluated using blocker antibodies for TLR2 and 4, peptide binding for TLR2 and 4, and stable shRNA TLR4 in the presence or not of aPL.
Methods: Patient information, blood serum and decidual tissue from gestationally aged-matched (8-10 weeks) women undergoing elective pregnancy terminations were used to determine BMI, serum C-reactive protein (CRP) (Qg/mL) and immune cell composition. Women were categorized as healthy BMI (20-24.9) not presenting with low-grade inflammation (CRP < 2 Qg/mL; ControlCRP-) or obese BMI ( 30) presenting with low-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 by multicolour flow cytometry analysis. Results: We did not detect differences in proportions of CD56bright decidual NKs, dendritic cells, macrophages or Th17 and CD8+ T cells between decidual tissues of ControlCRP- (n¼15) and ObeseCRP+ (n¼15) women. However, proportions of immunosuppressive CD4+/FoxP3+ Tregs, pro-inflammatory IFN©+ Th1 cells, and cytotoxic CD16+ NK cells were elevated in ObeseCRP+ decidua while IL-13+ Th2 proportions were decreased. Conclusion: This study suggests that obesity-linked inflammation in early pregnancy does alter immune cell composition within the maternal-fetal interface. These new findings lay the groundwork for future studies aimed at dissecting the importance of these cellular changes in pregnancy.
P1.151. PHAGOCYTIC ACTIVITY OF MACROPHAGES IS REGULATED BY FIRST TRIMESTER DECIDUAL CELLS IN RESPONSE TO PRO-INFLAMMATORY STIMULI VIA THE MODULATION OF “DON'T-EAT-ME” SIGNAL Longzhu Piao a, Susheela Tridandapani b, S. Joseph Huang a a Deartment of Ob/Gyn, The Ohio State University, Columbus, OH, USA; b SBS-Microbial Infection/Immunity, The Ohio State University, Columbus, OH, USA
A58
Abstracts / Placenta 35 (2014) A1eA112
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 the mechanisms of FTDC/macrophage interactions resulting in previously undisclosed homeostasis on placentation. Methods: FTDCs were primed with estradiol (E) (108 M) + medroxyprogesterone acetate (M) (107 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-1b- or 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 a, Moriya Gamliel b, Ronit Gilad a, Tal Imbar a, Ronit Haimov-Kochman a, Caryn Greenfield a, Iris Eisenberg-Loebl a, Ofer Mandelboim b, 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 (P¼0.0008 (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 finding may 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 rio-França b, Eduardo Glilciane Morceli a, Cristiane Hara b, Adenilda Hono bora Danny Laura Fagundes a, Marilza Rudge a, De França b, ~o Damasceno a, Iracema Calderon a a Botucatu Medical School, Botucatu/Sa Paulo, Brazil; b Institute of Biological and Health Science - Federal University of Mato Grosso, Barra do Garças/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 ¼ 15), mild hyperglycemic (N ¼ 15) and diabetic (N ¼ 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. Key words: cord blood, maternal blood, cytokines, diabetes, flow cytometry. Acknowledgements - FAPESP (2012/18033-0)
P1.154. IMMUNE SYSTEM AND REPRODUCTIVE SUCCESS: THE GENETICS OF THE MATERNAL-FETAL INTERFACE Olympe Chazara a, Annettee Nakimuli b, Lydia Farrell a, Susan Hiby a, Hugo Hilton c, Paul Norman c, Neda Nemat-Gorgani c, Alison Eliott d, e, Pontiano Florence Mirembe b, Peter Parham c, Ashley Kaleebu d, Moffett a a University of Cambridge, Cambridge, UK; b Makerere University, Kampala, Uganda; c Stanford University School of Medicine, Stanford, USA; d Medical Research Council/Uganda Virus Research Institute, Entebbe, Uganda; e London School of Hygiene and Tropical Medicine, London, UK Objectives: The success of human pregnancy depends on interactions between maternal killer cell immunoglobulin-like receptors (KIR) and their fetal ligands, HLA-C1 and C2. Our previous genetic studies of pregnancies show that in disorders of pregnancy due to defective placentation (pre-eclampsia, recurrent miscarriage and fetal growth restriction) there is an increased frequency of maternal KIRAA genotype in combination with a paternally derived fetal HLA-C2 group. The telomeric KIRB region provides protection from these disorders. Methods: Firstly, the inhibitory KIR2DL1, located in the centromeric KIR region, binds strongly to HLA-C2, whereas the activating receptor for HLAC2, KIR2DS1, is found on the telomeric B region. We have typed for allelic variation of KIR2DL1 and KIR2DS1 in our pregnancy cohorts (1300 pregnancies comparing controls and pre-eclampsia or recurrent miscarriage). Secondly, we have now conducted a case-control study in Kampala, Uganda. Mother-baby pairs from 484 normal pregnancies and 254 complicated with pre-eclampsia were genotyped for KIR and HLA-C1 or C2 groups. Result: We show that KIR2DL1 is polymorphic, with 11 alleles observed whereas KIR2DS1 is monomorphic. One of the common KIR2DL1 alleles, KIR2DL1*003, is associated with pre-eclampsia and recurrent miscarriage (P-value¼0.0016, OR 1.5 and P¼0.0125, OR 1.7 respectively).
Ă
B) Proton Magnetic resonance spectroscopy was performed to evaluate the biochemical/ metabolic profile in placentas. A significant decrease in phosphocholine 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). Decreased glucose levels and increased lactate levels, indicative of hypoxia, were measured 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). Placental insufficiency in APS was also associated with abnormal fetal neurodevelopment characterised by cortical axonal cytoarchitecture disruption(NF200 and MAP-2 expression) and increased neurodegeneration (FluoroJadeB). Increased anxiety was observed in the offspring of APS-mice.
A57
After determining the role of TLR4 in trophoblastic cell fusion impairment by aPL, signalling cascade of inflammation was studied by ELISA measuring TNFa and IL8 secretion. ER stress activation was assessed by qPCR and WB quantifying Glucose-regulated protein 78 (GRP78) and C/EBP-homologous protein (CHOP) expressions. Result: Fusion index and hCG secretion are decreased by addition of aPL. In the presence of aPL and anti-TLR4 antibodies or in BeWo cells transfected with shRNA TLR4, trophoblastic cell fusion and hCG secretion were restored. Neither TNFa nor IL8 secretion was observed in BeWo cells stimulated by aPL. Increase in GRP78 and CHOP expressions was observed in cells stimulated by aPL, but not when blocked with anti-TLR4 antibodies, suggesting a role of ER stress in trophoblastic cell fusion impairment by aPL. Conclusion: aPL interfered with fusion process and biochemical differentiation of trophoblastic cells via TLR4 that triggers ER stress activation.
P1.150. EXAMINING THE EFFECT OF OBESITY-ASSOCIATED SYSTEMIC INFLAMMATION ON THE UTERINE IMMUNE CELL NICHE IN EARLY PREGNANCY Yoona Kim, Sofie Perdu, Mahroo Aghababaei, Alexander Beristain The University of British Columbia, Vancouver, Canada Objectives: Low-grade inflammation associates with major obstetrical disorders like preterm birth and preeclampsia and is thought to be causal in the development of these high-risk pregnancies. Control of immune cell-directed pro-inflammatory cytokine production and cytotoxicity is crucial for utero-placental function; aberrant pro-inflammatory T helper (h) 1 cell activity within the uterine microenvironment associates with placental dysfunction and poor pregnancy outcome. Immunological imbalances and low-grade inflammation are associated with excess adiposity. However, the cellular mechanism(s) linking obesity-associated inflammation to poor pregnancy outcome are poorly understood. As a first step in addressing this knowledge gap, our study aims to examine multiple immune cell subsets within decidual tissues of non-obese and obese women in early pregnancy.
Ă
Conclusion: Using two novel non invasive in uterus MRI-based methods we were able to determine complement activation and metabolism in the placentas and predict fetal outcomes.
P1.149-N. PATHOLOGICAL MECHANISMS OF ANTIPHOSPHOLIPID ANTIBODIES IN TROPHOBLASTIC CELL FUSION Tess Marchetti a, b, Philippe de Moerloose b, Marie Cohen a a Laboratory of Hormonology, Geneva University Hospitals, Geneva, Switzerland; b Haemostasis Unit, Geneva University Hospitals, Geneva, Switzerland Objectives: Obstetrical Antiphospholipid Syndrome (APS) associates pregnancy pathologies with antiphospholipid antibodies (aPL). It has been suggested that aPL could affect trophoblastic cell fusion. However, their pathogenicity is still poorly understood. Toll-like Receptors (TLR) have been implicated in the pathological activation of aPL on endothelial cells. Although inflammation has been incriminated in this process, endoplasmic reticulum (ER) stress activation seems more related to trophoblastic fusion impairment by aPL. The aim of our study was to evaluate the pathological mechanisms of aPL on trophoblastic cell fusion. Methods: BeWo cell line is used as a model to mimic trophoblastic cell fusion, triggered by forskolin. Results were corroborated with primary cytotrophoblastic cells. Fusion index was determined by immunocytochemistry and biochemical differentiation by ELISA measuring hCG secretion. Effects of TLR on BeWo cell fusion were evaluated using blocker antibodies for TLR2 and 4, peptide binding for TLR2 and 4, and stable shRNA TLR4 in the presence or not of aPL.
Methods: Patient information, blood serum and decidual tissue from gestationally aged-matched (8-10 weeks) women undergoing elective pregnancy terminations were used to determine BMI, serum C-reactive protein (CRP) (Qg/mL) and immune cell composition. Women were categorized as healthy BMI (20-24.9) not presenting with low-grade inflammation (CRP < 2 Qg/mL; ControlCRP-) or obese BMI ( 30) presenting with low-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 by multicolour flow cytometry analysis. Results: We did not detect differences in proportions of CD56bright decidual NKs, dendritic cells, macrophages or Th17 and CD8+ T cells between decidual tissues of ControlCRP- (n¼15) and ObeseCRP+ (n¼15) women. However, proportions of immunosuppressive CD4+/FoxP3+ Tregs, pro-inflammatory IFN©+ Th1 cells, and cytotoxic CD16+ NK cells were elevated in ObeseCRP+ decidua while IL-13+ Th2 proportions were decreased. Conclusion: This study suggests that obesity-linked inflammation in early pregnancy does alter immune cell composition within the maternal-fetal interface. These new findings lay the groundwork for future studies aimed at dissecting the importance of these cellular changes in pregnancy.
P1.151. PHAGOCYTIC ACTIVITY OF MACROPHAGES IS REGULATED BY FIRST TRIMESTER DECIDUAL CELLS IN RESPONSE TO PRO-INFLAMMATORY STIMULI VIA THE MODULATION OF “DON'T-EAT-ME” SIGNAL Longzhu Piao a, Susheela Tridandapani b, S. Joseph Huang a a Deartment of Ob/Gyn, The Ohio State University, Columbus, OH, USA; b SBS-Microbial Infection/Immunity, The Ohio State University, Columbus, OH, USA
A58
Abstracts / Placenta 35 (2014) A1eA112
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 the mechanisms of FTDC/macrophage interactions resulting in previously undisclosed homeostasis on placentation. Methods: FTDCs were primed with estradiol (E) (108 M) + medroxyprogesterone acetate (M) (107 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-1b- or 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 a, Moriya Gamliel b, Ronit Gilad a, Tal Imbar a, Ronit Haimov-Kochman a, Caryn Greenfield a, Iris Eisenberg-Loebl a, Ofer Mandelboim b, 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 (P¼0.0008 (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 finding may 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 rio-França b, Eduardo Glilciane Morceli a, Cristiane Hara b, Adenilda Hono bora Danny Laura Fagundes a, Marilza Rudge a, De França b, ~o Damasceno a, Iracema Calderon a a Botucatu Medical School, Botucatu/Sa Paulo, Brazil; b Institute of Biological and Health Science - Federal University of Mato Grosso, Barra do Garças/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 ¼ 15), mild hyperglycemic (N ¼ 15) and diabetic (N ¼ 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. Key words: cord blood, maternal blood, cytokines, diabetes, flow cytometry. Acknowledgements - FAPESP (2012/18033-0)
P1.154. IMMUNE SYSTEM AND REPRODUCTIVE SUCCESS: THE GENETICS OF THE MATERNAL-FETAL INTERFACE Olympe Chazara a, Annettee Nakimuli b, Lydia Farrell a, Susan Hiby a, Hugo Hilton c, Paul Norman c, Neda Nemat-Gorgani c, Alison Eliott d, e, Pontiano Florence Mirembe b, Peter Parham c, Ashley Kaleebu d, Moffett a a University of Cambridge, Cambridge, UK; b Makerere University, Kampala, Uganda; c Stanford University School of Medicine, Stanford, USA; d Medical Research Council/Uganda Virus Research Institute, Entebbe, Uganda; e London School of Hygiene and Tropical Medicine, London, UK Objectives: The success of human pregnancy depends on interactions between maternal killer cell immunoglobulin-like receptors (KIR) and their fetal ligands, HLA-C1 and C2. Our previous genetic studies of pregnancies show that in disorders of pregnancy due to defective placentation (pre-eclampsia, recurrent miscarriage and fetal growth restriction) there is an increased frequency of maternal KIRAA genotype in combination with a paternally derived fetal HLA-C2 group. The telomeric KIRB region provides protection from these disorders. Methods: Firstly, the inhibitory KIR2DL1, located in the centromeric KIR region, binds strongly to HLA-C2, whereas the activating receptor for HLAC2, KIR2DS1, is found on the telomeric B region. We have typed for allelic variation of KIR2DL1 and KIR2DS1 in our pregnancy cohorts (1300 pregnancies comparing controls and pre-eclampsia or recurrent miscarriage). Secondly, we have now conducted a case-control study in Kampala, Uganda. Mother-baby pairs from 484 normal pregnancies and 254 complicated with pre-eclampsia were genotyped for KIR and HLA-C1 or C2 groups. Result: We show that KIR2DL1 is polymorphic, with 11 alleles observed whereas KIR2DS1 is monomorphic. One of the common KIR2DL1 alleles, KIR2DL1*003, is associated with pre-eclampsia and recurrent miscarriage (P-value¼0.0016, OR 1.5 and P¼0.0125, OR 1.7 respectively).