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173 REPEATED IMPLANTATION FAILURE. DIAGNOSTICS AND TREATMENT OPTIONS
Immunology of Reproduction 172 THE STUDY OF IMMUNOSUPPRESSIVE EFFECT OF PREGNANT MOUSE SERUM ON DENDRITIC CELLS S.M. Moazzeni1 , J. Shojaeian1 , S. Nikoo1 , M. Bozorgmehr1 , A.H. Zarnani2 . 1 Department of Immunology, Tarbiat Modares University, Tehran, Iran, 2 Immunology Research Center, Iran University of Medical Sciences, Tehran, Iran Introduction: In normal pregnancy the maternal immune system is directed towards tolerance or suppression in order to prevent the rejection of the semi allogenic fetus. Antigen presenting cells especially dendritic cells (DCs) are the key cells in initiation and regulation of immune responses. The presence of potent immunostimulatory dendritic cells in the decidual tissue of pregnancy has been demonstrated. The aim of this study was to determine how allostimulatory activity of DCs could be affected during pregnancy. Materials and Methods: DCs were isolated from spleen of pregnant or non-pregnant Balb/c mice and co-cultured with allogenic C57BL/6 T lymphocytes prepared by nylon wool method from brachial lymph nodes. Some cultures of non-pregnant female DCs were treated by 2.5% serum obtained from pregnant mice at early, middle or late gestational periods and were used in the same mixed lymphocyte reaction (MLR) settings. Cell proliferation was measured by 3 H thymidin incorporation and cytokine production was measured in supernatants of MLR cultures using ELISA method. Effect of pregnant mouse serum on expression of DC surface markers was evaluated by flow cytometry. Results: No significant difference was found between stimulatory potential of splenic DCs from pregnant and non-pregnant mice in induction of allogenic T cell proliferative response. Moreover, serum of early or late pregnancy did not have any effect on DCs function in comparison with non-pregnant mouse serum, while mid-pregnancy serum significantly inhibited allostimulatory activity of DCs. IFNg production in co-culture of DCs treated with pregnant mouse serum was significantly lower than that of control group, however, no significant difference in IL-10 production was observed. Treatment of DCs with pregnant mouse serum did not influence the percentage of cells expressing MHCII, CD86, CD8a or CD11b. However a marked reduction of the mean fluorescence intensity of MHC-II was observed. Conclusion: Our results concerning the diminished capacity of DCs to induce production of TH1 cytokines and allogenic T cell proliferation after treatment with pregnant mouse serum reveals a new way of immunologic tolerance against semi-allogenic fetus. 173 REPEATED IMPLANTATION FAILURE. DIAGNOSTICS AND TREATMENT OPTIONS V.I. Pirohova1 , L.O. Mykhaylyshyn2 . 1 The Lviv National Medical University after Danylo Halytskyi, Department of Obstetrics and Gynaecology, Lviv, Ukraine, 2 Medical Center “Intersono”, Department of Reproductive Endocrinology, Lviv, Ukraine Introduction: Assisted reproductive technologies have been developed considerably since the first in vitro fertilization (IVF) birth in 1978, but pregnancy rates have not increased significantly during the last years. The lack of increase in pregnancy rate can be mainly attributed to implantation failure, the causes of which remain elusive, thus hindering the establishment of adequate treatment. The aim of the study was to propose a set of tests to clarify the diagnosis of repeated implantation failure (RIF) in patients
S77 undergoing IVF and to evaluate effectiveness of the provided treatment in case of pathology detection. Materials and Methods: 89 non-pregnant patients up to 35 years old, without any uterine pathology and paternal karyotype abnormalities (detected in both spouses), with at least three unsuccessful IVF attempts were included into the survey. These previous IVF attempts were characterised by transferring blastocysts of high quality on 5 day development. All these patients were divided into two groups. Group I was comprised of 48 women who after informed consent, were tested for the presence of antiphospholipid (aPL), antithyroid antibodies (ATA), increased levels of circulating natural killer (NK)-cells, T helper 1 predisposition, inherited thrombophilia, serum homocysteine level. Group I was divided into 2 subgroups: IA with detected pathology and appropriate treatment (intravenous immunoglobulin (IVIg), vitamins B, folic acid, low dosage aspirin, heparin, clopidogrel) simultaneously with IVF program, IB without pathology, who were provided with conventional IVF. Group II, 41 women underwent conventional further attempt of IVF without mentioned above examinations and treatment. Results and Discussion: 72.9% of patients had at least one abnormal result out of all the tests performed. The immunological changes were the most frequently detected pathology with the rate of 68.75% for women experiencing RIF. The second place was taken by ATA and they were found in 22.9% of the patients. The presence of aPL was detected in 6.3% of women experiencing RIF. The conclusion has been drawn, that all of ATA and aPL positive patients had elevated NK-cells. The prevalence of inherited thrombophilic abnormalities in group of patients with RIF was the same as in average health population. Pregnancy rate in Group IA, IB, II of patients was 77.1%, 7.7%, 12.2% respectively. The difference appeared to be statistically significant (p < 0.05). Conclusions: The most frequent abnormal changes were elevated blood NK cells, T helper 1 predisposition within the group of RIF patients with transferring high quality embryos. The prevalence of inherited thrombophilic abnormalities was the same as in average health population. It has been proved that IVIg therapy is a method of high effectiveness in case of immunological pathology treatment. 174 SYSTEMIC INFLAMMATORY CONDITION AS A CAUSE OF ART PREGNANCY COMPLICATIONS T.T. Saroyan, I.E. Korneeva, T.A. Nazarenko, G.T. Sukhikh. Obstetrics, Gynecology and Perinatology Research Center, Department of Reproduction, Moscow, Russia Development of severe OHSS involves two important pathophysiological mechanisms: (1) activation of angiogenesis and increase in vascular permeability partly due to VEGF effects; (2) systemic inflammatory response syndrome (SIRS) which is associated with massive endothelial damage and release of proinflammatory cytokines and acute phase proteins into circulation. Objective: To assess severe OHSS contribution to further pregnancy complications. Materials and Methods: Group 1 (n = 70) included patients with ART pregnancies and OHSS, group 2 (n = 30) patients with ART pregnancies without OHSS, control group 15 patients with spontaneous pregnancies without complications. Serum concentrations of VEGF, proinflammatory cytokines (IL-2, IL-6, TNF-a), acute phase proteins (C-reactive protein (CRP), fibrinogen, von Willebrandt
S77 undergoing IVF and to evaluate effectiveness of the provided treatment in case of pathology detection. Materials and Methods: 89 non-pregnant patients up to 35 years old, without any uterine pathology and paternal karyotype abnormalities (detected in both spouses), with at least three unsuccessful IVF attempts were included into the survey. These previous IVF attempts were characterised by transferring blastocysts of high quality on 5 day development. All these patients were divided into two groups. Group I was comprised of 48 women who after informed consent, were tested for the presence of antiphospholipid (aPL), antithyroid antibodies (ATA), increased levels of circulating natural killer (NK)-cells, T helper 1 predisposition, inherited thrombophilia, serum homocysteine level. Group I was divided into 2 subgroups: IA with detected pathology and appropriate treatment (intravenous immunoglobulin (IVIg), vitamins B, folic acid, low dosage aspirin, heparin, clopidogrel) simultaneously with IVF program, IB without pathology, who were provided with conventional IVF. Group II, 41 women underwent conventional further attempt of IVF without mentioned above examinations and treatment. Results and Discussion: 72.9% of patients had at least one abnormal result out of all the tests performed. The immunological changes were the most frequently detected pathology with the rate of 68.75% for women experiencing RIF. The second place was taken by ATA and they were found in 22.9% of the patients. The presence of aPL was detected in 6.3% of women experiencing RIF. The conclusion has been drawn, that all of ATA and aPL positive patients had elevated NK-cells. The prevalence of inherited thrombophilic abnormalities in group of patients with RIF was the same as in average health population. Pregnancy rate in Group IA, IB, II of patients was 77.1%, 7.7%, 12.2% respectively. The difference appeared to be statistically significant (p < 0.05). Conclusions: The most frequent abnormal changes were elevated blood NK cells, T helper 1 predisposition within the group of RIF patients with transferring high quality embryos. The prevalence of inherited thrombophilic abnormalities was the same as in average health population. It has been proved that IVIg therapy is a method of high effectiveness in case of immunological pathology treatment. 174 SYSTEMIC INFLAMMATORY CONDITION AS A CAUSE OF ART PREGNANCY COMPLICATIONS T.T. Saroyan, I.E. Korneeva, T.A. Nazarenko, G.T. Sukhikh. Obstetrics, Gynecology and Perinatology Research Center, Department of Reproduction, Moscow, Russia Development of severe OHSS involves two important pathophysiological mechanisms: (1) activation of angiogenesis and increase in vascular permeability partly due to VEGF effects; (2) systemic inflammatory response syndrome (SIRS) which is associated with massive endothelial damage and release of proinflammatory cytokines and acute phase proteins into circulation. Objective: To assess severe OHSS contribution to further pregnancy complications. Materials and Methods: Group 1 (n = 70) included patients with ART pregnancies and OHSS, group 2 (n = 30) patients with ART pregnancies without OHSS, control group 15 patients with spontaneous pregnancies without complications. Serum concentrations of VEGF, proinflammatory cytokines (IL-2, IL-6, TNF-a), acute phase proteins (C-reactive protein (CRP), fibrinogen, von Willebrandt