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Thyroid hormone receptors and reproduction
Abstracts / Journal of Reproductive Immunology 86 (2010) 79–111
83
Funded by the grants from the Natural Sciences and Engineering Research Council of Canada.
9
doi:10.1016/j.jri.2010.08.008
Ralf Dittrich
8
OB/GYN, University Hospital Erlangen, University of ErlangenNuremberg, Germany
The immunosuppressive role of the atrial natriuretic peptide (ANP) in preovulatory human follicle Dineva a,∗ ,
Vangelov a ,
J. I. Nikolov b , M.D. Ivanova a
V.
Terzieva a ,
D.
Gulenova b ,
G.
a Institute of Biology and Immunology of Reproduction « Acad.K.Bratanov », Bulgaria b ReproBiomed Center, Sofia, Bulgaria
Introduction: ANP and its receptor, membrane guanylyle cyclase, are actively involved in cGMP signalling during folliculogenesis and ovulation. Based on our previous studies on the production and localization of ANP in preovulatory human follicles, we aimed to investigate the immunoregulatory role of ANP and the relationship of ANP concentration in follicular fluids (FFls) with COH/IVF results and granulosa cells apoptosis. Methods: TNF␣ and IFN␥ were measured by human TNF␣ and IFN␥ kits (eBioscience, CA, USA) in LPS stimulated and ANP treated human whole blood cell culture. The concentration of ANP in FFls in infertile women was measured by proANP kit (BIOMEDICA, Austria) and its relationship with COH/IVF results (number of punctured follicles, number of isolated oocytes and number of fertilized oocytes) was analyzed statistically (One way ANOVA, Correlation matrices). The antiapoptotic effect of ANP was studied in in vitro culture of Granulosa luteinized cells (GLCs) by the measure of Caspase-3 activity (Ac-DEVD-PNA). Results: The immunosuppressive effect of the low concentrations of 1.3–1.6 × 10−12 M ANP (measured in preovulatory human FFls) was found in LPS stimulated whole human blood cell culture by the inhibition of TNF␣ and IFN␥ production. The significant positive correlation among the ANP concentration in FFls and the number of aspirated follicles (r = 0.4476, P < 0.05), number of isolated (r = 0.5785, P < 0.05) and number of fertilized oocytes (r = 0.5973, P < 0.05) was demonstrated in COH/IVF patients. The rhANP as well as the induced synthesis of ANP inhibited Caspase-3 activity in hGLC culture, P > 0.05. Conclusions: The immunosuppressive and survival role of ANP was demonstrated in human preovulatory follicle. A positive correlation between the ANP concentration in FFls with the COH/IVF results was found in studied patients. Keywords: Granulosa cells; ANP; Apoptosis; Cytokines; COH/IVF Funding source: National Sci. Fund B 1507/05 doi:10.1016/j.jri.2010.08.009
Thyroid hormone receptors and reproduction
Thyroid disorders have a great impact on fertility in both sexes. Hyper- and hypothyroidism cause changes in sex hormone binding globulin (SHBG), prolactin, gonadotropin releasing hormone and sex steroids serum levels. In females, there is also an assumed possible direct effect of thyroid hormones on oocytes due to the fact that specific binding sites for thyroxin have been identified in mouse and human oocytes (Krassas et al., 2010). Thyroid dysfunction in females has also been associated with pregnancy morbidity and pregnancy outcome. In males, hyperthyroidism causes a reduction of sperm motility and hypothyroidism increases the count of abnormal sperm according to the morphology. Both abnormalities improve or normalize when euthyroidism is restored. In females, alterations of the fertility by thyroid disorders are more complex. Hyper- and hypothyroidism are the main thyroid diseases that have an adverse effect on female reproduction and causes menstrual disturbances, mainly hypomenorrhea and polymenorrhea in hyperthyroidism and oligomenorrhea in hypothyroidsm. In recent studies it has become evident that not only changes in serum levels of SHBG and sex steroids are responsible for these disorders, but also alterations of the metabolic pathway. In hypothyroidism, glucose uptake in muscle and adipose tissue is resistant to insulin, resulting in higher levels of insulin in these patients. Whereas these changes are well recognized in patients with hypothyroidism, the evidence regarding such a relationship in subclinical hypothyroidism is inconsistent. Mueller et al. evaluated the association between thyroid function, reflected by thyroidstimulating hormone (TSH) levels, and insulin resistance (IR) in 337 women suffering from polycystic ovary syndrome (PCOS) and found a significant association between thyroid function, as reflected by TSH ≥2 mIU/l, and IR (Mueller et al., 2009). Today there is no doubt that subclinical hypothyroidism must be treated by application with thyroxin in infertile women. Additionally, thyroid autoimmunity is involved in infertility. Thyroid autoimmunity is the most common endocrine disorder in women of reproductive age. 5–20 women out of 100 are affected. Most studies dealing with this topic showed an increased prevalence of thyroid autoimmunity in women attending infertility centres (Poppe et al., 2008). Autoimmune thyroid disorders are characterized by the presence of thyroid auto-antibodies, particularly thyroid peroxidase autoantibodies (TPO-Ab) and anti-thyroglobulin autoantibodies (TG-AB). Many women with severe hypothyroidism (basal serum TSH >4.5 mIU/l) had thyroid antibodies in a study by Grassi et al. (2001). Patients with thyroid antibodies and hypothyroidism must be distinguished from patients with thyroid antibodies and euthyroidism. The presence of thyroid antibodies alone does not influence the pregnancy rates (Poppe et al., 2007). In clinical praxis, infertile women
84
Abstracts / Journal of Reproductive Immunology 86 (2010) 79–111
should be screened for thyroid disorders when the male is not identified for being the cause of infertility. When clinical or subclinical thyroid hypo activity is detected (TSH >2.5 mIU/l), thyroxin should be administered. In the case of polycystic ovary syndrome patients, thyroxin should be administered even when TSH is over 2.0 mIU/l. Hyperthyroidsm should be treated by antithyroidals or surgery. The presence of thyroid antibodies alone is not responsible for a lower pregnancy rate. When these women get pregnant, they should be further controlled and treated when necessary. References Grassi, G., et al., 2001. Trends Mol. Med. 15:, 389. Krassas, G.E., et al., 2010. Endocr. Rev. 31 (epub ahead of print). Mueller, A., et al., 2009. Hum. Reprod. 24, 2924. Poppe, K., et al., 2008. Nat. Clin. Pract. 4, 394. Poppe, K., et al., 2007. Clin. Endocr. 66, 309.
doi:10.1016/j.jri.2010.08.010 10 Circulating cytokines influence the fetal growth in pregnant women with rheumatoid arthritis R.J.E.M. Dolhain a,b,∗ , F.D.O. de Steenwinkel a,b , Y.A. de Man a,b , A.C.S. Hokken- Koelega a , Y.B. de Rijke b , J.M.W. Hazes a,b a
Deps. of Rheumatology, Paediatric Endocrinology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands b Deps. of Clinical Chemistry, Erasmus MC, University Medical Center, Rotterdam, The Netherlands Introduction: High rheumatoid arthritis (RA) disease activity during pregnancy is associated with lower birth weight. Lower birth weight, even within the normal range, has been linked with cardiovascular diseases and metabolic syndromes later in life. Why high disease activity results in lower birth weight is not known. Outside pregnancy, active RA is associated with high levels of circulating cytokines. We therefore aim to assess the serum levels of IL-10, IL-6 and TNF␣ in pregnant women with RA and to evaluate correlations between these levels, disease activity, and birth weight. Methods: Current study is embedded in the PARAstudy, a prospective study on RA and pregnancy. 134 pregnant RA patients are enrolled in first and 168 in third trimester. 33 healthy pregnant women served as control. We analysed data using birth weight standard deviation scores (bwsds). Maternal disease activity is based on the disease activity score for 28 joints (DAS28) (range 0–10). IL-10, IL-6 and TNF␣ were determined on an Immulite 1000 system. Results: In RA patients levels of IL-10, IL-6 and TNF␣ decrease during pregnancy, most likely as a results of declining disease activity. Strong correlations were found between these cytokines in first and third trimester and between DAS28 and IL-10, IL-6. First trimester patients with detectable IL-10 (n = 12) showed a higher disease activity than the IL-10 negative patients, a mean DAS28 4.4 (SD 1.2) and 3.6 (SD 1.1) respec-
tively. To diminish the influence of disease activity on birth weight we matched the IL-10 positive to an IL-10 negative group. Matching was on disease activity, parity and prednisone use. Mean bwsds was significantly higher (p = 0.02) in the IL-10 positive group 0.92 (SD 0.7) than in the IL10 negative match, 0.15 (SD 0.7). No such association was found in third trimester. To determine the additional effect of IL-6 to disease activity on bwsds stratification was done. In first trimester we stratified IL-6 and DAS28 in a high and low group based on their median, resulting in 4 groups. In the two high DAS28 groups (defined median over 3.8), bwsds was significantly lower in high IL-6 than in low IL-6 (p < 0.05). In the high and low IL-6 groups bwsds was −0.19 (SD 1.12) and 0.36 (SD 0.93), respectively. No such association was found in third trimester. TNF␣ was stratified the same way. In third trimester, only in patients with low disease activity, bwsds was lower in the low TNF␣ group than in the high TNF␣ group, bwsds of 0.05 (SD 0.97) and 0.52 (SD 0.96) respectively (p < 0.05). Same trend was seen in the healthy cohort group, although not significant. No such association was found in first trimester or in patients with high RA disease activity. Conclusions: Circulating cytokines influence fetal growth in pregnant women with RA. In first trimester elevated IL-10 seems to protect against the negative influence of RA disease activity on birth weight, IL-6 seems to amplify this negative influence. Both cytokines create a bwsds deviation of more than 0.50 which is considered clinically relevant. In third trimester there is no influence suggesting an early critical window. Finally, our data might indicate a physiological involvement of TNF␣ in fetal growth during third trimester in healthy controls and patients with low disease activity. This implicates that TNF-blockers, which are more and more prescribed during pregnancy to treat RA, should be used with caution. Funding source: Dutch Arthritis Association doi:10.1016/j.jri.2010.08.011 11 Allergen induced release of immune mediators in human placentae—A possible role in fetal programming of allergies? U. Enke ∗ , J. Heinzelmann, V. Varosi, M. Schleussner, U.R. Markert, L. Seyfarth
Abelius, E.
Placenta Labor, Department of Obstetrics, Friedrich-SchillerUniversität Jena, Germany Introduction: Previous animal experiments indicated the transmissibility of general allergic diathesis from the mother to the fetus. All major cell types known to be involved in allergic processes, including dendritic cells, T cells, B cells and mast cells are present in the decidua. Thus, allergen induced mediator release may prime the fetus for easier sensitization in later life. Material and methods: To investigate possible differences in cytokine and chemokine production from
83
Funded by the grants from the Natural Sciences and Engineering Research Council of Canada.
9
doi:10.1016/j.jri.2010.08.008
Ralf Dittrich
8
OB/GYN, University Hospital Erlangen, University of ErlangenNuremberg, Germany
The immunosuppressive role of the atrial natriuretic peptide (ANP) in preovulatory human follicle Dineva a,∗ ,
Vangelov a ,
J. I. Nikolov b , M.D. Ivanova a
V.
Terzieva a ,
D.
Gulenova b ,
G.
a Institute of Biology and Immunology of Reproduction « Acad.K.Bratanov », Bulgaria b ReproBiomed Center, Sofia, Bulgaria
Introduction: ANP and its receptor, membrane guanylyle cyclase, are actively involved in cGMP signalling during folliculogenesis and ovulation. Based on our previous studies on the production and localization of ANP in preovulatory human follicles, we aimed to investigate the immunoregulatory role of ANP and the relationship of ANP concentration in follicular fluids (FFls) with COH/IVF results and granulosa cells apoptosis. Methods: TNF␣ and IFN␥ were measured by human TNF␣ and IFN␥ kits (eBioscience, CA, USA) in LPS stimulated and ANP treated human whole blood cell culture. The concentration of ANP in FFls in infertile women was measured by proANP kit (BIOMEDICA, Austria) and its relationship with COH/IVF results (number of punctured follicles, number of isolated oocytes and number of fertilized oocytes) was analyzed statistically (One way ANOVA, Correlation matrices). The antiapoptotic effect of ANP was studied in in vitro culture of Granulosa luteinized cells (GLCs) by the measure of Caspase-3 activity (Ac-DEVD-PNA). Results: The immunosuppressive effect of the low concentrations of 1.3–1.6 × 10−12 M ANP (measured in preovulatory human FFls) was found in LPS stimulated whole human blood cell culture by the inhibition of TNF␣ and IFN␥ production. The significant positive correlation among the ANP concentration in FFls and the number of aspirated follicles (r = 0.4476, P < 0.05), number of isolated (r = 0.5785, P < 0.05) and number of fertilized oocytes (r = 0.5973, P < 0.05) was demonstrated in COH/IVF patients. The rhANP as well as the induced synthesis of ANP inhibited Caspase-3 activity in hGLC culture, P > 0.05. Conclusions: The immunosuppressive and survival role of ANP was demonstrated in human preovulatory follicle. A positive correlation between the ANP concentration in FFls with the COH/IVF results was found in studied patients. Keywords: Granulosa cells; ANP; Apoptosis; Cytokines; COH/IVF Funding source: National Sci. Fund B 1507/05 doi:10.1016/j.jri.2010.08.009
Thyroid hormone receptors and reproduction
Thyroid disorders have a great impact on fertility in both sexes. Hyper- and hypothyroidism cause changes in sex hormone binding globulin (SHBG), prolactin, gonadotropin releasing hormone and sex steroids serum levels. In females, there is also an assumed possible direct effect of thyroid hormones on oocytes due to the fact that specific binding sites for thyroxin have been identified in mouse and human oocytes (Krassas et al., 2010). Thyroid dysfunction in females has also been associated with pregnancy morbidity and pregnancy outcome. In males, hyperthyroidism causes a reduction of sperm motility and hypothyroidism increases the count of abnormal sperm according to the morphology. Both abnormalities improve or normalize when euthyroidism is restored. In females, alterations of the fertility by thyroid disorders are more complex. Hyper- and hypothyroidism are the main thyroid diseases that have an adverse effect on female reproduction and causes menstrual disturbances, mainly hypomenorrhea and polymenorrhea in hyperthyroidism and oligomenorrhea in hypothyroidsm. In recent studies it has become evident that not only changes in serum levels of SHBG and sex steroids are responsible for these disorders, but also alterations of the metabolic pathway. In hypothyroidism, glucose uptake in muscle and adipose tissue is resistant to insulin, resulting in higher levels of insulin in these patients. Whereas these changes are well recognized in patients with hypothyroidism, the evidence regarding such a relationship in subclinical hypothyroidism is inconsistent. Mueller et al. evaluated the association between thyroid function, reflected by thyroidstimulating hormone (TSH) levels, and insulin resistance (IR) in 337 women suffering from polycystic ovary syndrome (PCOS) and found a significant association between thyroid function, as reflected by TSH ≥2 mIU/l, and IR (Mueller et al., 2009). Today there is no doubt that subclinical hypothyroidism must be treated by application with thyroxin in infertile women. Additionally, thyroid autoimmunity is involved in infertility. Thyroid autoimmunity is the most common endocrine disorder in women of reproductive age. 5–20 women out of 100 are affected. Most studies dealing with this topic showed an increased prevalence of thyroid autoimmunity in women attending infertility centres (Poppe et al., 2008). Autoimmune thyroid disorders are characterized by the presence of thyroid auto-antibodies, particularly thyroid peroxidase autoantibodies (TPO-Ab) and anti-thyroglobulin autoantibodies (TG-AB). Many women with severe hypothyroidism (basal serum TSH >4.5 mIU/l) had thyroid antibodies in a study by Grassi et al. (2001). Patients with thyroid antibodies and hypothyroidism must be distinguished from patients with thyroid antibodies and euthyroidism. The presence of thyroid antibodies alone does not influence the pregnancy rates (Poppe et al., 2007). In clinical praxis, infertile women
84
Abstracts / Journal of Reproductive Immunology 86 (2010) 79–111
should be screened for thyroid disorders when the male is not identified for being the cause of infertility. When clinical or subclinical thyroid hypo activity is detected (TSH >2.5 mIU/l), thyroxin should be administered. In the case of polycystic ovary syndrome patients, thyroxin should be administered even when TSH is over 2.0 mIU/l. Hyperthyroidsm should be treated by antithyroidals or surgery. The presence of thyroid antibodies alone is not responsible for a lower pregnancy rate. When these women get pregnant, they should be further controlled and treated when necessary. References Grassi, G., et al., 2001. Trends Mol. Med. 15:, 389. Krassas, G.E., et al., 2010. Endocr. Rev. 31 (epub ahead of print). Mueller, A., et al., 2009. Hum. Reprod. 24, 2924. Poppe, K., et al., 2008. Nat. Clin. Pract. 4, 394. Poppe, K., et al., 2007. Clin. Endocr. 66, 309.
doi:10.1016/j.jri.2010.08.010 10 Circulating cytokines influence the fetal growth in pregnant women with rheumatoid arthritis R.J.E.M. Dolhain a,b,∗ , F.D.O. de Steenwinkel a,b , Y.A. de Man a,b , A.C.S. Hokken- Koelega a , Y.B. de Rijke b , J.M.W. Hazes a,b a
Deps. of Rheumatology, Paediatric Endocrinology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands b Deps. of Clinical Chemistry, Erasmus MC, University Medical Center, Rotterdam, The Netherlands Introduction: High rheumatoid arthritis (RA) disease activity during pregnancy is associated with lower birth weight. Lower birth weight, even within the normal range, has been linked with cardiovascular diseases and metabolic syndromes later in life. Why high disease activity results in lower birth weight is not known. Outside pregnancy, active RA is associated with high levels of circulating cytokines. We therefore aim to assess the serum levels of IL-10, IL-6 and TNF␣ in pregnant women with RA and to evaluate correlations between these levels, disease activity, and birth weight. Methods: Current study is embedded in the PARAstudy, a prospective study on RA and pregnancy. 134 pregnant RA patients are enrolled in first and 168 in third trimester. 33 healthy pregnant women served as control. We analysed data using birth weight standard deviation scores (bwsds). Maternal disease activity is based on the disease activity score for 28 joints (DAS28) (range 0–10). IL-10, IL-6 and TNF␣ were determined on an Immulite 1000 system. Results: In RA patients levels of IL-10, IL-6 and TNF␣ decrease during pregnancy, most likely as a results of declining disease activity. Strong correlations were found between these cytokines in first and third trimester and between DAS28 and IL-10, IL-6. First trimester patients with detectable IL-10 (n = 12) showed a higher disease activity than the IL-10 negative patients, a mean DAS28 4.4 (SD 1.2) and 3.6 (SD 1.1) respec-
tively. To diminish the influence of disease activity on birth weight we matched the IL-10 positive to an IL-10 negative group. Matching was on disease activity, parity and prednisone use. Mean bwsds was significantly higher (p = 0.02) in the IL-10 positive group 0.92 (SD 0.7) than in the IL10 negative match, 0.15 (SD 0.7). No such association was found in third trimester. To determine the additional effect of IL-6 to disease activity on bwsds stratification was done. In first trimester we stratified IL-6 and DAS28 in a high and low group based on their median, resulting in 4 groups. In the two high DAS28 groups (defined median over 3.8), bwsds was significantly lower in high IL-6 than in low IL-6 (p < 0.05). In the high and low IL-6 groups bwsds was −0.19 (SD 1.12) and 0.36 (SD 0.93), respectively. No such association was found in third trimester. TNF␣ was stratified the same way. In third trimester, only in patients with low disease activity, bwsds was lower in the low TNF␣ group than in the high TNF␣ group, bwsds of 0.05 (SD 0.97) and 0.52 (SD 0.96) respectively (p < 0.05). Same trend was seen in the healthy cohort group, although not significant. No such association was found in first trimester or in patients with high RA disease activity. Conclusions: Circulating cytokines influence fetal growth in pregnant women with RA. In first trimester elevated IL-10 seems to protect against the negative influence of RA disease activity on birth weight, IL-6 seems to amplify this negative influence. Both cytokines create a bwsds deviation of more than 0.50 which is considered clinically relevant. In third trimester there is no influence suggesting an early critical window. Finally, our data might indicate a physiological involvement of TNF␣ in fetal growth during third trimester in healthy controls and patients with low disease activity. This implicates that TNF-blockers, which are more and more prescribed during pregnancy to treat RA, should be used with caution. Funding source: Dutch Arthritis Association doi:10.1016/j.jri.2010.08.011 11 Allergen induced release of immune mediators in human placentae—A possible role in fetal programming of allergies? U. Enke ∗ , J. Heinzelmann, V. Varosi, M. Schleussner, U.R. Markert, L. Seyfarth
Abelius, E.
Placenta Labor, Department of Obstetrics, Friedrich-SchillerUniversität Jena, Germany Introduction: Previous animal experiments indicated the transmissibility of general allergic diathesis from the mother to the fetus. All major cell types known to be involved in allergic processes, including dendritic cells, T cells, B cells and mast cells are present in the decidua. Thus, allergen induced mediator release may prime the fetus for easier sensitization in later life. Material and methods: To investigate possible differences in cytokine and chemokine production from