- Email: [email protected]
GnRH I is expressed by CD14+ monocytes and dendritic cells
80
Abstracts / Journal of Reproductive Immunology 94 (2012) 5–130
pregnancy loss of genetically impaired embryos found later on. Methods and patients: Sera of these patients contained high levels of IgG antibodies against cardiolipin, IgG and IgM against phospahtidyl inositol, IgG against phospahatidyl L-serin, IgG antibodies against annexin V examined by ELISA. Convetional treatment of antiphospholipid syndrome was ineffective. RESULTS: Preimplantation genetic diagnosis (PGD) showed chromosomally impaired embryos. Relationships between disease activities of the severity of antiphospholipid syndrome in the both patients were similar and the treatment was directed according to the clinical symptoms and laboratory results. CONCLUSION: Pregnancies and deliveries were recorded due to the close collaboration of in vitro fertilization and diagnosis of early embryos. Both women displayed primarily the signs of antiphospholipid syndrome (positive laboratory results, frequent miscarriages). After the selection of embryos, using the method of PGD, both patients became pregnant and delivered healthy fetuses. Without such selection, both women would have probably aborted their embryos again. Supported by a grant MSM 002 162 0812
three months of TNFi administrations, during the luteal phase. Th1 and Th2 cells were determined by detecting the intracellular TNF-␣, IFN-␥ and IL-10 production, using flow cytometer. Data were analyzed by non-parametric paired samples Wilcoxon test. Diffe-rences were considered significant ifP< 0.05. RESULTS: The percentages of TNF-␣-producing Th cells and IFN-␥-producing Th cells were significantly reduced (38.03% ± 11.40%VS30.16% ± 10.47%,P = 0.000, and 25.34% ± 8.74%VS19.64% ± 7.91%,P = 0.000, respectively) whereas the percentage of IL-10-producing Th cells had no change (1.62% ± 0.86%VS1.48% ± 0.76%,P = 0.198) after TNFi administrations compared those before TNFi administrations. The Th1/Th2 ratios of TNF-␣/IL-10 and IFN-␥/IL-10 were significantly reduced (27.70 ± 13.46VS22.94 ± 9.89,P = 0.000, and 18.44 ± 9.33VS15.06 ± 7.55,P = 0.000, respectively) after TNFi administrations compared those before TNFi administrations. CONCLUSIONS: Tumor necrosis factor inhibitors suppress the express of Th1 cytokines and shift the peripheral Th1/Th2 ratio from Th1 dominant status to Th2 dominant status, which may be benefit for embryo implantation and development in uRM orRIFpatients.
doi:10.1016/j.jri.2012.03.387
doi:10.1016/j.jri.2012.03.388
P 086
P 087
Tumor Necrosis Factor Inhibitors Tip the Th Balance in Women with Unexplained Recurrent Miscarriage or Repeated Implantation Failure
GnRH I is expressed by CD14+ monocytes and dendritic cells
G.-G. Li 1,2,3,∗ , P. Liang 1,2,3 , J. Cai 1,2,3 , X.-Y. Chen 1,2,3 , X.D. Hu 1,2,3 , M.-L. Mo 1,2,3 , C.-P. Guo 1,2,3 , B. Yin 1,2,3 , Y. Zeng 1,2,3 , X.-H. Liao 1,2,3 , D.-M. Zhang 1,2,3 1
Shenzhen Zhongshan Urology Hospital, Clinical Center for Recurrent Miscarriage & Recurrent Implantation Failure, Shenzhen, China 2 Shenzhen Zhongshan Urology Hospital, Assisted Reproductive Technology Center, Shenzhen, China 3 Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics, Shenzhen, China PROBLEM: Several recently publications reported that tumor necrosis factor inhibitors (TNFi) were benefit for embryo implantation and pregnant maintenance. However, information regarding its mechanisms remains scarce. The objective of the study was to evaluate the alterations in the proportion of peripheral TNF-␣, IFN-␥ or IL-10 producing Th cells and the Th1/Th2 ratios of TNF-␣/IL-10 and IFN-␥/IL-10 in patients with unexplained recurrent miscarriage (uRM) or repeated implantation failure (RIF) before and after TNFi administrations. METHODS: Eighty-nine patients with a history of uRM orRIFand with elevated endometrial CD57 cells or elevated peripheral TNF-␣/IL-10 ratio were included in the study. Either Adalimumab 40 mg was injected by subcutaneous injection every 2 weeks or Etanercept 25 mg every 96 hr. Patients’ peripheral bleed were sampled prior to the first administration of TNFi and sampled again after two or
S.E. Segerer ∗ , M. Kapp, J. Dietl, U. Kämmerer University of Wuerzburg, Obstetrics and Gynecology, Wuerzburg, Germany PROBLEM: With the onset of pregnancy, a significant increase of antigen-presenting cells (APC), especially dendritic cells (DC) is observed. DC represent a highly adaptive cell type. Without antigen contact or in a special antiinflammatory micromilieu, DC exhibit an immuninhibitory phenotype, which can inhibit stimulatory T-cells. Adopting this special skill, DC could have a major impact on the establishment of tolerance against the semiallogenic fetus during pregnancy. So far, only the effects of several glycoprotein hormones and steroidhormones on DC were investigated. No investigation dealt with the impact and function of gonadotropin-releasing hormone (GnRH), the key regulator of reproductive functions secreted from the hypothalamus. We therefore intended to investigate the expression of its corresponding receptor GnRHI on decidual immune cell populations (CD56+ uterine natural killer cells, CD14+ monocytes, DC), on decidual stromal cells and on invasive cytotrophoblast cells. METHOD: Investigation of the GnRH receptor expression in human decidual tissue (7-8 weeks of gestation) was performed by Western Blot. Therefore, decidual leukocytes, i.e. CD56 + +CD16- uterine NK cells (uNK), CD14+ monocytes as well as cytotrophoblast (CTB) and stromal cells were isolated by positive selection.
Abstracts / Journal of Reproductive Immunology 94 (2012) 5–130
RESULTS: After selection of decidual cell subsets, we observed that GnRH I is expressed by CD 14+ monocytes, DC and CTB. No expression was found in CD56 + +CD16- uNK. CONCLUSION: Expressing GnRH I, DC could be a suitable target for the action of the superordinate regulator GnRH to influence the immune responses during early pregnancy. doi:10.1016/j.jri.2012.03.389 P 088 Sex-specific effect of first trimester maternal progesterone on birth weight I. Hartwig 1,∗ , M. Pincus 2 , A. Diemert 1 , K. Hecher 1 , P. Arck 1 1 University Medical Center Hamburg-Eppendorf, Department of Obstetrics and Fetal Medicine, Hamburg, Germany 2 Charité University Hospital Berlin, Department of Pediatrics, Berlin, Germany
PROBLEM: Birth weight is a strong predictor for shortterm survival of newborns and long-term health outcome. Low birth weight increases a child’s risk to develop diseases, such as cardiovascular disease, type 2 diabetes and asthma. Though macro- and micro-environmental risk factors affecting birth weight have been identified, e. g. maternal stress or smoking, biological mediators affecting or predicting birth weight alterations remain elusive. Possible biological mediators determining fetal growth and birth weight may derive from the maternal immune and endocrine system. These systems are pivotal in maternal adaptation to pregnancy and hence, fetal development. E.g., progesterone is essential to promoting the immunological adaptation. Emerging evidence indicates that progesterone levels can be affected by environmental challenges, such as stress and smoking. We aimed to determine whether maternal progesterone in early pregnancy affects birth weight and is sensitive to environmental insults, such as stress and smoking during first trimester of pregnancy in a fetal-sex dependent manner. METHODS: Pregnant women were recruited through obstetricians in private practice and enrolled in a prospective cohort study at the Charité University Medicine in Berlin, Germany. Anthropometric, medical and psychosocial information, including perceived stress, was collected and hormone levels were measured in 906 women during first trimester of pregnancy, followed by documentation of pregnancy outcome after birth. Univariate and multivariate regression analyses were performed to identify maternal markers, among them progesterone, affecting birth weight. Further, regression analyses were used to determine which environmental and maternal factors affect maternal progesterone level during pregnancy. RESULTS: Maternal progesterone levels during first trimester were positively associated with birth weight at term in girls, but not boys. After correcting for other variables, each increase in progesterone by 1 ng/ml increased girls’ birth weight by 8.93 g (95% confidence interval, 1.00
81
to 16.85 g). If the mother carried a boy, maternal smoking during early pregnancy predicted reduced birth weight, irres-pective of progesterone levels. Multivariate analysis revealed that a maternal body mass index (BMI) over 25 and maternal age under 21 years significantly correlated with reduced levels of progesterone. Correlations between macro- and microenvironmental challenges and maternal progesterone levels did not reach levels of significance. CONCLUSIONS: Maternal progesterone in the first trimester is a good indicator of fetal growth in women pregnant with a girl, but not with a boy. Further investigations into the mechanisms of progesterone are needed to determine why this effect is sex-specific. Also, detailed identification of macro- and micro-environmental factors modulating maternal progesterone levels should be addressed in future studies. We propose that screening of maternal progesterone levels may become a valuable tool to predict pregnancies at risk to result in the birth of a girl with low birth weight. doi:10.1016/j.jri.2012.03.390 P 089 Detection of circulating cell-derived microparticles, about the methodology A.-R. Han 1,∗ , R.W. Dickinson 2 , A. Gilman-Sachs 2 , J. KwakKim 1,2 1
Reproductive Medicine, The Chicago Medical School at Rosalid Franklin University, Obstetrics and Gynecology, Vernon Hills, United States 2 The Chicago Medical School at Rosalind Franklin University, Microbiology and Immunology, North Chicago, United States PROBLEM: Circulating cell-derived microparticles (MPs) have been implicated in several disease processes including adverse pregnancy outcomes such as pregnancy loss and preeclampsia. However, the studies are limited due to a lack of standardization in its isolation technique and accurate measurement. In this study we aimed to investigate the current method to isolate MPs and multiple markers for detecting various MPs including trophoblast-derived MPs (tMPs). METHODS: Blood was sampled from non-pregnant and pregnant women with history of infertility or recurrent pregnancy losses. Circulating MPs in plasma of were isolated by centrifugation at 1,550 g for 15 min at 20 ◦ C first, and then 13,000 g for 3 min at 10 ◦ C within 4 hours after sample collection. MPs were characterized using multiple monoclonal antibodies such as anti-CD41, andti-CD31, anti-CD144, anti-CD45, anti-HLA-G and anti-a2V-ATPase, anti-CD235a, and annexin V and determined its origin (i.e. platelet, endothelial cell, leukocyte, and trophoblast). Various factors which may affect the MP isolation such as blood collecting tube, fresh vs. frozen sample, markers for tMPs and others on MP measurement were investigated. RESULTS: Application of citrate tube for collecting blood samples decreased the total count of MPs compared to heparin tube. Anti-HLA-G and anti-a2V-ATPase, which are
Abstracts / Journal of Reproductive Immunology 94 (2012) 5–130
pregnancy loss of genetically impaired embryos found later on. Methods and patients: Sera of these patients contained high levels of IgG antibodies against cardiolipin, IgG and IgM against phospahtidyl inositol, IgG against phospahatidyl L-serin, IgG antibodies against annexin V examined by ELISA. Convetional treatment of antiphospholipid syndrome was ineffective. RESULTS: Preimplantation genetic diagnosis (PGD) showed chromosomally impaired embryos. Relationships between disease activities of the severity of antiphospholipid syndrome in the both patients were similar and the treatment was directed according to the clinical symptoms and laboratory results. CONCLUSION: Pregnancies and deliveries were recorded due to the close collaboration of in vitro fertilization and diagnosis of early embryos. Both women displayed primarily the signs of antiphospholipid syndrome (positive laboratory results, frequent miscarriages). After the selection of embryos, using the method of PGD, both patients became pregnant and delivered healthy fetuses. Without such selection, both women would have probably aborted their embryos again. Supported by a grant MSM 002 162 0812
three months of TNFi administrations, during the luteal phase. Th1 and Th2 cells were determined by detecting the intracellular TNF-␣, IFN-␥ and IL-10 production, using flow cytometer. Data were analyzed by non-parametric paired samples Wilcoxon test. Diffe-rences were considered significant ifP< 0.05. RESULTS: The percentages of TNF-␣-producing Th cells and IFN-␥-producing Th cells were significantly reduced (38.03% ± 11.40%VS30.16% ± 10.47%,P = 0.000, and 25.34% ± 8.74%VS19.64% ± 7.91%,P = 0.000, respectively) whereas the percentage of IL-10-producing Th cells had no change (1.62% ± 0.86%VS1.48% ± 0.76%,P = 0.198) after TNFi administrations compared those before TNFi administrations. The Th1/Th2 ratios of TNF-␣/IL-10 and IFN-␥/IL-10 were significantly reduced (27.70 ± 13.46VS22.94 ± 9.89,P = 0.000, and 18.44 ± 9.33VS15.06 ± 7.55,P = 0.000, respectively) after TNFi administrations compared those before TNFi administrations. CONCLUSIONS: Tumor necrosis factor inhibitors suppress the express of Th1 cytokines and shift the peripheral Th1/Th2 ratio from Th1 dominant status to Th2 dominant status, which may be benefit for embryo implantation and development in uRM orRIFpatients.
doi:10.1016/j.jri.2012.03.387
doi:10.1016/j.jri.2012.03.388
P 086
P 087
Tumor Necrosis Factor Inhibitors Tip the Th Balance in Women with Unexplained Recurrent Miscarriage or Repeated Implantation Failure
GnRH I is expressed by CD14+ monocytes and dendritic cells
G.-G. Li 1,2,3,∗ , P. Liang 1,2,3 , J. Cai 1,2,3 , X.-Y. Chen 1,2,3 , X.D. Hu 1,2,3 , M.-L. Mo 1,2,3 , C.-P. Guo 1,2,3 , B. Yin 1,2,3 , Y. Zeng 1,2,3 , X.-H. Liao 1,2,3 , D.-M. Zhang 1,2,3 1
Shenzhen Zhongshan Urology Hospital, Clinical Center for Recurrent Miscarriage & Recurrent Implantation Failure, Shenzhen, China 2 Shenzhen Zhongshan Urology Hospital, Assisted Reproductive Technology Center, Shenzhen, China 3 Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics, Shenzhen, China PROBLEM: Several recently publications reported that tumor necrosis factor inhibitors (TNFi) were benefit for embryo implantation and pregnant maintenance. However, information regarding its mechanisms remains scarce. The objective of the study was to evaluate the alterations in the proportion of peripheral TNF-␣, IFN-␥ or IL-10 producing Th cells and the Th1/Th2 ratios of TNF-␣/IL-10 and IFN-␥/IL-10 in patients with unexplained recurrent miscarriage (uRM) or repeated implantation failure (RIF) before and after TNFi administrations. METHODS: Eighty-nine patients with a history of uRM orRIFand with elevated endometrial CD57 cells or elevated peripheral TNF-␣/IL-10 ratio were included in the study. Either Adalimumab 40 mg was injected by subcutaneous injection every 2 weeks or Etanercept 25 mg every 96 hr. Patients’ peripheral bleed were sampled prior to the first administration of TNFi and sampled again after two or
S.E. Segerer ∗ , M. Kapp, J. Dietl, U. Kämmerer University of Wuerzburg, Obstetrics and Gynecology, Wuerzburg, Germany PROBLEM: With the onset of pregnancy, a significant increase of antigen-presenting cells (APC), especially dendritic cells (DC) is observed. DC represent a highly adaptive cell type. Without antigen contact or in a special antiinflammatory micromilieu, DC exhibit an immuninhibitory phenotype, which can inhibit stimulatory T-cells. Adopting this special skill, DC could have a major impact on the establishment of tolerance against the semiallogenic fetus during pregnancy. So far, only the effects of several glycoprotein hormones and steroidhormones on DC were investigated. No investigation dealt with the impact and function of gonadotropin-releasing hormone (GnRH), the key regulator of reproductive functions secreted from the hypothalamus. We therefore intended to investigate the expression of its corresponding receptor GnRHI on decidual immune cell populations (CD56+ uterine natural killer cells, CD14+ monocytes, DC), on decidual stromal cells and on invasive cytotrophoblast cells. METHOD: Investigation of the GnRH receptor expression in human decidual tissue (7-8 weeks of gestation) was performed by Western Blot. Therefore, decidual leukocytes, i.e. CD56 + +CD16- uterine NK cells (uNK), CD14+ monocytes as well as cytotrophoblast (CTB) and stromal cells were isolated by positive selection.
Abstracts / Journal of Reproductive Immunology 94 (2012) 5–130
RESULTS: After selection of decidual cell subsets, we observed that GnRH I is expressed by CD 14+ monocytes, DC and CTB. No expression was found in CD56 + +CD16- uNK. CONCLUSION: Expressing GnRH I, DC could be a suitable target for the action of the superordinate regulator GnRH to influence the immune responses during early pregnancy. doi:10.1016/j.jri.2012.03.389 P 088 Sex-specific effect of first trimester maternal progesterone on birth weight I. Hartwig 1,∗ , M. Pincus 2 , A. Diemert 1 , K. Hecher 1 , P. Arck 1 1 University Medical Center Hamburg-Eppendorf, Department of Obstetrics and Fetal Medicine, Hamburg, Germany 2 Charité University Hospital Berlin, Department of Pediatrics, Berlin, Germany
PROBLEM: Birth weight is a strong predictor for shortterm survival of newborns and long-term health outcome. Low birth weight increases a child’s risk to develop diseases, such as cardiovascular disease, type 2 diabetes and asthma. Though macro- and micro-environmental risk factors affecting birth weight have been identified, e. g. maternal stress or smoking, biological mediators affecting or predicting birth weight alterations remain elusive. Possible biological mediators determining fetal growth and birth weight may derive from the maternal immune and endocrine system. These systems are pivotal in maternal adaptation to pregnancy and hence, fetal development. E.g., progesterone is essential to promoting the immunological adaptation. Emerging evidence indicates that progesterone levels can be affected by environmental challenges, such as stress and smoking. We aimed to determine whether maternal progesterone in early pregnancy affects birth weight and is sensitive to environmental insults, such as stress and smoking during first trimester of pregnancy in a fetal-sex dependent manner. METHODS: Pregnant women were recruited through obstetricians in private practice and enrolled in a prospective cohort study at the Charité University Medicine in Berlin, Germany. Anthropometric, medical and psychosocial information, including perceived stress, was collected and hormone levels were measured in 906 women during first trimester of pregnancy, followed by documentation of pregnancy outcome after birth. Univariate and multivariate regression analyses were performed to identify maternal markers, among them progesterone, affecting birth weight. Further, regression analyses were used to determine which environmental and maternal factors affect maternal progesterone level during pregnancy. RESULTS: Maternal progesterone levels during first trimester were positively associated with birth weight at term in girls, but not boys. After correcting for other variables, each increase in progesterone by 1 ng/ml increased girls’ birth weight by 8.93 g (95% confidence interval, 1.00
81
to 16.85 g). If the mother carried a boy, maternal smoking during early pregnancy predicted reduced birth weight, irres-pective of progesterone levels. Multivariate analysis revealed that a maternal body mass index (BMI) over 25 and maternal age under 21 years significantly correlated with reduced levels of progesterone. Correlations between macro- and microenvironmental challenges and maternal progesterone levels did not reach levels of significance. CONCLUSIONS: Maternal progesterone in the first trimester is a good indicator of fetal growth in women pregnant with a girl, but not with a boy. Further investigations into the mechanisms of progesterone are needed to determine why this effect is sex-specific. Also, detailed identification of macro- and micro-environmental factors modulating maternal progesterone levels should be addressed in future studies. We propose that screening of maternal progesterone levels may become a valuable tool to predict pregnancies at risk to result in the birth of a girl with low birth weight. doi:10.1016/j.jri.2012.03.390 P 089 Detection of circulating cell-derived microparticles, about the methodology A.-R. Han 1,∗ , R.W. Dickinson 2 , A. Gilman-Sachs 2 , J. KwakKim 1,2 1
Reproductive Medicine, The Chicago Medical School at Rosalid Franklin University, Obstetrics and Gynecology, Vernon Hills, United States 2 The Chicago Medical School at Rosalind Franklin University, Microbiology and Immunology, North Chicago, United States PROBLEM: Circulating cell-derived microparticles (MPs) have been implicated in several disease processes including adverse pregnancy outcomes such as pregnancy loss and preeclampsia. However, the studies are limited due to a lack of standardization in its isolation technique and accurate measurement. In this study we aimed to investigate the current method to isolate MPs and multiple markers for detecting various MPs including trophoblast-derived MPs (tMPs). METHODS: Blood was sampled from non-pregnant and pregnant women with history of infertility or recurrent pregnancy losses. Circulating MPs in plasma of were isolated by centrifugation at 1,550 g for 15 min at 20 ◦ C first, and then 13,000 g for 3 min at 10 ◦ C within 4 hours after sample collection. MPs were characterized using multiple monoclonal antibodies such as anti-CD41, andti-CD31, anti-CD144, anti-CD45, anti-HLA-G and anti-a2V-ATPase, anti-CD235a, and annexin V and determined its origin (i.e. platelet, endothelial cell, leukocyte, and trophoblast). Various factors which may affect the MP isolation such as blood collecting tube, fresh vs. frozen sample, markers for tMPs and others on MP measurement were investigated. RESULTS: Application of citrate tube for collecting blood samples decreased the total count of MPs compared to heparin tube. Anti-HLA-G and anti-a2V-ATPase, which are