Psychological stress, immune system and IVF outcome

Abstracts / Journal of Reproductive Immunology 81 (2009) 113–175

vated variants (CD56+dim /CD16+/CD161+/NKG-2D+). Peripheral blood exerted increased CD85j surface expression on T helper cells (CD3+/CD4+/CD8−), bearing NK cell activation marker NKG-2D in the case of successfully fertilized patients. These results indicate the important role of KIR2DL4 mediated attenuation of NK cell function in the scope of IVF success rate. Increased CD85j (ILT2 or LIR) expression on T helper cells may have contributed to overall immune attenuation and IVF success. (Supported by IGA Grant NR/9135-3, Ministry of Health of the Czech Republic.) doi:10.1016/j.jri.2009.06.218 P13 Markers associated with success and failure of embryo transfer (ET) during IVF program D.Yu. Trofimov, O.V. Burmenskaya, M.V. Ebzeeva, E.A. Kalinina, S.M. Mullabaeva, L.N. Kuzjmichev, G.T. Sukhikh Scientific Center for Obstetrics, Gynecology and Perinatology named after V.I. Kulakov, Moscow, Russia In terms of IVF-ET program we have investigated cytokines mRNA expression profiles and bacterial contamination background in endometrial tissue as well as vaginal flora content by means of qPCR. The subjects were women (n = 23) with tubo-peritoneal factor of infertility. The following cytokines were analyzed in the study with normalization to “housekeeping genes” (HPRT1, TBP, B2 M) using Cp method: IL-1␤, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12a/p35, IL-12b/IL23/p40, IL-15, IL-17, IL-18, IL-23/p19, TNF-␣, TGF-␤, IFN-␥, Foxp3, LIF, and ␣-chain IL-2R. The quantity of total bacteria, Lactobacillus spp. and 14 major groups of microorganisms comprising vaginal flora were evaluated by the presence of 16S rRNA. Endometrium samples were obtained at proliferation phase (cycle day 7–9) prior to ovarian stimulation as well as during “implantation window”. Vaginal swabs were taken on 7–9th day of cycle. By the moment an embryo transfer was performed for 9 women and resulted in 2 positive outcomes, 6 negative ones, and a single case of ectopic pregnancy. From the obtained data we consider as negative factors for ET increased expression of the proinflammatory cytokines: IL-1␤ (Me 570 vs 5), IL-8 (Me 1660 vs 22), TNF-␣ (Me 24 vs 5), IL-17 (Me 28 vs 1), and IL-18 (Me 8.9 vs 3.3). Comparing cytokine profiles at various phases of cycle we have found increased IL-2 mRNA level in endometrial proliferation phase against that in “implantation window” (Me 57.7 vs 6.5). High expression

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level of proinflammatory cytokines were correlated with increased total bacteria count in endometrium (10–100 times) consisting of non-Lactobacillus flora. As markers associated with positive implantation outcome we suggest moderate expression of proinflammatory cytokines, significant expression of LIF and TGF-␤ in a certain range and no bacterial contamination of endometrium. doi:10.1016/j.jri.2009.06.219 P14 Psychological stress, immune system and IVF outcome K. Gourounti a,f , V. Kapetanios b , N. Paparisteidis c , G. Vaslamatzis d , F. Anagnostopoulos e a

Department of Midwifery, TEI of Athens, Greece

b Department of Infertility, ‘Locus Medicus’ Medical and

Research Center, Athens, Greece c Department of Assisted Reproduction, Helena Venizelou Hospital, Athens, Greece d Department of Psychoanalytic Psychotherapy, Medical School of University of Athens, Eginition Hospital, Athens, Greece e Department of Psychology, Panteion University, Athens, Greece f Postnatal and Antinatal Department, Helena Venizelou Hospital, Athens, Greece Psychological distress has become a commonly sited factor when discussing reproductive failures. Psychological stress may act through different plausible mechanisms on the reproductive system. Three systems – the endocrine, immune and nervous systems – engage in multiple interactions with the body’s response to acute and chronic stress. The hypothalamic-pituitary-adrenal (HPA) axis, when activated by stress, exerts an inhibitory effect on the female reproductive system. Corticotropin-releasing hormone inhibits GnRH secretion, and glucocorticoids inhibit LH and ovarian estrogen and progesterone secretion. An acute stress during the follicular phase may lead to a premature LH surge and thereby interfere with follicular maturation and ovulation. Furthermore, stress trigger an intense local inflammatory response and simultaneously suppress the systemic inflammatory response. The intensive local inflammatory response may be linked to follicular atresia and luteolysis. The aim of this review was to present the available evidence regarding the possible impact of psychological stress on IVF outcome. A broad search of the psychological and medical electronic databases (Medline, PsycINFO, CINAHL, and EMBASE) was undertaken at the begin-

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Abstracts / Journal of Reproductive Immunology 81 (2009) 113–175

ning of 2008 with the keywords, infertility, fertility related distress, stress, in vitro fertilization. Twenty-eight studies approximately, have investigated the impact of distress on IVF outcome. Two-thirds of these studies have found a significant relationship between infertility distress and subsequent pregnancy rates. Major findings from these studies showed that the baseline and procedural levels of anxiety (acute and chronic) and depression, mood state and certain coping modes were significantly associated to numerous biological end-points, including number of oocytes retrieved and fertilized and pregnancy rate. The remaining studies either noticed a trend or did not find a relationship between distress and pregnancy outcome. There are many studies that support that psychological stress may impact on IVF outcome and specifically decreases the pregnancy rates. It is possible that treatments could combine hormone and psychological interventions. doi:10.1016/j.jri.2009.06.220 P15 Cellular immunity in the outcome of in vitro fertilization–embryo transfer V. Kitsiou b , S. Dendrinos b , K. Psarra a , V. Kapsimali a , G. Creatsas b , C. Papasteriades a a

Immunology and Histocompatibility Department, “Evangelismos” Hospital, Athens, Greece b B’ Obstetrics and Gynecology Department, University of Athens, Athens, Greece The close relationship of reproductive outcome and immune system has been recognized many years ago and has been extensively studied. In vitro fertilization (IVF) and embryo transfer (ET) are well-established techniques overcoming the problem of infertility. The rate of IVF/ET success remains low, with an elusive etiology in most cases. The aim of this study was to evaluate the involvement of cellular immunity in women with multiple failures of IVF/ET attempts, through the assessment of peripheral blood (PB) leukocyte subpopulations. The study included 55 infertile women with failed IVF treatments and 45 fertile women used as a control group. The immunophenotyping of the PB leukocyte subpopulations (CD2+, CD3+, CD3+CD4+, CD3+CD8+, CD5+CD19+, CD3−CD16/56+, CD3+TCR␥␦+, CD3−CD16+, CD3−CD56+, CD3−CD16+CD56+, CD3−CD16−CD56+) was performed in whole blood by flow cytometry. The study showed (A) a statistically significant increase CD3+TCR␥␦+ (p = 0.018), CD3−CD56+ (p = 0.005) and CD3−CD16+CD56+

(p < 0.001) cells in the study group in comparison to the control group, (B) no statistically significant difference in the CD2+, CD3+, CD3+CD4+, CD3+ CD8+, CD5+CD19+, CD3−CD16/56+, CD3−CD16+, CD3−CD16−CD56+ subpopulations between the two groups studied. In conclusion, the increase of PB CD3+ TCR␥␦+, CD3−CD56+ and CD3−CD16+CD56+ cells, known to participate in the pathogenesis of recurrent spontaneous abortions as well, may affect negatively the outcome of IVF/ET. Therefore the study of these parameters seems to contribute substantially to the pre IVF/ET immunological evaluation and may further help in manipulations of these cases. doi:10.1016/j.jri.2009.06.221 P16 G-CSF pharmacologic supplementation in the ART (Assisted Reproductive Technologies) treatment cycles of low responder women F. Scarpellini a , M. Sbracia a , A. Patella b a

Hungaria Center for Endocrinology and Reproductive Medicine (CERM), Rome, Italy b University of Ferrara, Department of Ob/Gyn, Ferrara, Italy Granulocyte-Colony Stimulating Factor (G-CSF) is a cytokine discovered as a growth factor for granulocytes but it has been shown to have a lot of other physiologic actions. Recently, its role in the reproductive systems has been pointed out and particularly in ovarian follicles where the G-CSF concentration was found to be directly correlated to the oocyte quality and this was in relationship to the patient age. In all the techniques of ART (Assisted Reproductive Technology) oocyte number recruitment is crucial and a good quality of them is mandatory for the success of the event. The most important problem for every ART procedure is the low responder patients. In order to improve the response of the ovary to the pharmacologic stimulatory treatment we administered a supplementation of G-CSF to low responder women in ART cycles. A randomized controlled study was conducted in ART undergoing women referred to II level center for Reproductive Medicine between January 2000 till to June 2008 using G-CSF or placebo. The inclusion criteria were: woman’s age ranging between 35 and 43 years, patients who had got at least two low responder cycles with the same different protocols, and FSH ≥ 15 UI/L and they had to be negative for abnormal karyotype, congenital and acquired Mulleran defects, infections, endocrine problems, autoimmune