ABSTRACTS

SPEAKERS’ ABSTRACTS OPENING SESSION

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Global rates of preterm birth are high and those in the United States are among the highest in the economically opulent countries. Our failure to understand the basis of this problem can be traced to our substantial ignorance of antecedents normal labor and – an important corollary – whether preterm birth is merely the result of an early onset of normal labor, or an altogether different pathophysiologic process. Current research, approaching this question from a point of view of a variety of disciplines, in a manner than bars no concept or method of inquiry, but actually mandates interactive approaches and inquiries emanating from a host of disciplines stands a chance of unraveling this complex question. Details of this novel approach and its current application will be discussed and its implications reviewed.

powerful antibiotic peptides by the fat body. As an example, the basal level of expression of the antifungal peptide DROSOMYCIN, is increased a thousand fold within 30 minutes of septic injury in larvae or adults. In 1996, we discovered that the Toll signal transduction pathway, which was already known for its role in the setting up of the early dorso-ventral axis in the Drosophila embryo, controlled the antifungal response of the adult fly. This work was the breakthrough that led to the discovery of the human homologues of Toll, the Toll-Like-Receptors (TLRs). The paramount role of the TOLL pathway in the host defence of Drosophila is illustrated by experiments in which mutant flies are challenged with fungi or bacteria. In TOLL-deficient mutants, survival to fungal, but not to bacterial infection, is severely compromised. In vertebrates, recognition of microbes by the innate immune system takes place at the cellular level by the TLRs. In Drosophila however, these recognition events take place in the open circulatory system via soluble excreted recognition proteins. We will describe the immune system of Drosophila in an evolutionary and historical perspective.

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Prematurity, a global public health problem and a scientific puzzle M Katz March of Dimes, NY, USA

Inside the Nobel-Prize winning research on Toll-like receptors

Human endometrial receptivity – basic science to clinical applications

J-M Reichhart

M Ruiz, C Simo´n

University of Strasbourg and Institut Universitaire de France, France

n IVI (FIVI)- Universidad de Valencia, IVIOMICS, Spain Fundacio

During evolution, the adaptative immune system with its antibodies, B and T cells arose around 500 million years ago probably in the first vertebrates. This specific immune system acts in concert with the innate immune system in roughly 45,000 vertebrate species as a defence against invading microorganisms. In all invertebrates, the immune system is purely innate and this system represents the most ancient defence mechanism, already present in the first multicellular organisms. We have been working since 1985 on the innate immune system, using Drosophila as a model. In the fly, an infection provokes the rapid synthesis of

The endometrium is a hormonally regulated organ that is non-adhesive to embryos throughout most of the menstrual cycle in humans. Endometrial receptivity refers to a hormone-limited period in which the endometrial tissue acquires a functional and transient ovarian steroid-dependent status allowing blastocyst adhesion. Functional genomic studies of human endometrium in natural cycles have demonstrated that endometrial receptivity is an active process involving up- and down-regulation of hundreds of genes. Personalized medicine is a well-accepted concept in reproductive medicine except for the endometrial factor that is still neglected. Our group has developed

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the endometrial receptivity array (ERA), a customized array of 238 genes coupled to a computational predictor capable of diagnosing a functionally receptive endometrium regardless of its histological appearance. The accuracy of the diagnostic tool ERA has been demonstrated to be superior to endometrial histology and results are completely reproducible 29 to 40 months later. The aim of this presentation is to demonstrate the diagnostic and therapeutic efficiency of the ERA in patients with implantation failure (IF), through personalization of embryo transfer (pET). A multicenter prospective clinical study analyzing the results of ERA in 116 IF patients (5.7 previous failed cycles), either with their own oocytes (IVF) (n = 69) or ovum donation (OD) (n = 47). In non-receptive, ERA was repeated on the day indicated by the predictor, and pET was guided according to ERA results. Endometrial biopsies were collected at LH+7, or in HRT after 5 days of P impregnation. RNA was extracted and hybridized according to ERA technology. Data were analysed by the predictor using the SVM+KNN algorithm for the matrix of 238 genes and results were given as receptive (R) or non-receptive (NR). We propose for the first time the concept of pET in patients with IF guided by ERA as they have a displacement of the endometrial window of receptivity. This concept can be extended to patients in their first visit to the fertility clinic. Preliminary data suggest that around 15% of all patients can be NR. We are conducting a RCT comparing ERA personalized embryo transfer versus routine embryo transfer in patients undergoing ART.

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Pre-conceptual endometrial immune profiling: a new method to increase pregnancy rates in a context of previous unexplained and repeated embryo implantation failures after IVF/ICSI 1,2

1,2

1,2

N Le´de´e , M Petitbarat , K Vezmar , H Gahery2, A Bensussan2, G Chaouat2 ^pital Saint Louis, France; 2UMRS-976, MatriceLab Innove SARL, Ho  Paris-Diderot, Ho ^pital Saint Louis, France INSERM, Universite

1

Problem: A unique immune reaction occurs during the implantation window within the endometrium and is essential (i) to promote embryo adhesion and

(ii) to regulate the invasion phase. Disequilibrium of such a vital reaction may impede implantation. We developed a specific method documenting this local immune reaction, using pre-established criteria from a fertile cohort, to increase implantation through personalized therapeutic strategies aiming to neutralize the documented deregulation. Method of Study: A prospective cohort study included 267 women with history of repeated embryo implantation failures after IVF/ICSI (RIF). An endometrial biopsy was performed during a non-conceptual cycle. After standard datation, we quantified the uterine Natural killer cells (uNK) mobilization (immunostaining CD56+), the mRNA expression of interleukin-15 (uNK cells activation/maturation state), interleukin-18 (angiogenesis, Th-1/Th-2 balance) and TWEAK/Fn-14 (TNF related local immuno-regulation) by real time PCR. Treatments specific for either poor or over- endometrial immune activation were applied promote either the adhesion or to control the invasion steps. We assessed the treatment result by the ongoing pregnancy rate (PR) occurring after the first embryo(s) transfer (ET) following the evaluation. Results: In 86% (229/267), the endometrial profiles appeared to be deregulated according to the pre-established criteria. Endometrial over-immune activation was diagnosed amongst 62% (165/267) of women. Personalized strategy to avoid embryo loss resulted in a 39% ongoing PR (64/165). Conversely, 24% of women (64/267) displayed a low endometrial activation profile. Adhesion promoting personalized strategies resulted in a 50% ongoing PR (32/64). In 14% of the cases, we did not find and obvious deregulation (38/267) and the ongoing PR 16% (6/38), significantly lower than what is obtain in the deregulated and treated groups (P < 0.003). This corresponds to the PR usually observed in such RIF context. Conclusion: Endometrial immune profiling is useful for increasing implantation in a RIF context.

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Molecular networks in the embryo and endometrium for pregnancy success M Ho¨lker, D Salilew-Wondim, E Held, D Tesfaye, K Schellander Institute of Animal Science, University of Bonn, Germany

Problem: In the absence of sufficient knowledge of human pre-implantation embryo development, American Journal of Reproductive Immunology 71 (Suppl. 1) (2014) 15–28

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success rates of IVF have remained relatively low. Consequently, to improve the odds of pregnancy given the uncertainty regarding embryo developmental potential, multiple embryos are transferred in some clinics, resulting in multiple births causing some associated complications. Understanding the cellular and molecular mechanisms representing the early embryo-environmental crosstalk, however, could help to enlighten our knowledge and thus to increase the overall efficiency. However, owing to limited resources, the cellular and molecular mechanisms governing this early stage of human development are poorly understood so far. Method of Study: To provide insights into early mammalian developmental pathways, studies employing bovine embryo biopsies were conducted to connect molecular signatures determined by global gene expression analysis and subsequent embryo developmental capacities. Accordingly, global gene expression analysis employing endometrial samples categorized according to pregnancy outcome after embryo transfer were performed to identify molecular pathways related to endometrial receptivity. Results: Blastocysts with developmental competence to term displayed a typical molecular signature. Expression of tumor necrosis factor alpha (TNFa) was more abundant in blastocysts of low developmental capacity whereas abundance of beta-2-microglobulin (B2M) was higher in those bearing high developmental competence. With respect to endometrial receptivity, distinct molecular networks of the bovine endometrium related to cell to cell communication skills and inflammatory immune mediators have been identified to be correlated with improved receptivity and pregnancy success. Conclusion: The present work provides a comprehensive inventory of functional networks and genes expressed in the receptive bovine endometrium representing the maternal part of the embryo-maternal cross-talk. Likewise, large scale differences at the molecular level were identified for embryos associated with term developmental competence compared to those with lower developmental competence. Collectively, these results provide key insights into important developmental pathways of early mammalian embryos.

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Intralipid therapy for recurrent pregnancy loss – controversies and future directions J Braverman, DR Ritsick Braverman Reproductive Immunology PC, Woodbury, NY, USA

Intralipid is a 20% fat emulsion containing soybean oil triglycerides that was developed as a source of parenteral nutrition for patients unable to tolerate an oral diet. Immunomodulatory functions of Intralipid infusions were subsequently noted and Intralipid infusions, touted by many as an inexpensive alternative to IVIG, are now widely used to treat recurrent pregnancy loss. While Intralipid infusions have been used for years in women experiencing recurrent pregnancy loss with anecdotal success, a satisfactory explanation for its therapeutic effects has been largely lacking. We will discuss controversies regarding its potential immunological mechanisms of action. There are several studies that demonstrate inhibitory effects of Intralipid on NK cell cytotoxicity, but there is so far no satisfactory description of the mechanism by which this effect is achieved. It is also not clear if Intralipid-mediated suppression of NK cell cytotoxicity is a relevant mechanism for its effects on preventing immunological pregnancy loss. Alternatively, Intralipid’s effects may be mediated through metabolic effects on T cells. While resting T cells have a relatively low metabolic demand and use a balance of glucose, lipids, and amino acids as their metabolic fuel, activation of T cells causes them to undergo a dramatic metabolic reprogramming. While activation of effector T (Teff) cells of the Th1, Th2 and Th17 induces a decrease in fatty acid oxidation (FAO) and shift glucose metabolism away from oxidative phosphorylation and towards glycolysis, regulatory T (Treg) cells conversely rely heavily on FAO, and not on glycolysis, for fuel. The distinct metabolic differences between Treg and other T cell lineages may provide a target for selective immunomodulation that could be exploited therapeutically. Recent studies in fact have demonstrated that addition of exogenous fatty acids to T cells during activation inhibits differentiation of Teff cells and favors differentiation of Treg cells. Additionally, metformin increases Treg cell generation by binding to and activating AMP kinase, which in turn inhibits mTOR and causes a decrease in glycolysis and an increase in

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FAO. Thus, Intralipid infusion may function as one arm of an immunometabolic approach to promote Treg cell-mediated tolerance of the semi-allogenic embryo and efficient embryo implantation. While there are obvious differences in their biochemical composition, Intralipid infusions are promoted by many clinicians as an inexpensive and effective functional alternative to IVIG. We will discuss this controversial idea informed by both the literature and our extensive clinical experience with both Intralipid and IVIG, including evaluation of our own data. Finally, we will discuss areas for future research including the use of alternative intravenous fat emulsions containing higher ratios of x-3 to x-6 fatty acids.

NA responses can be induced by human seminal fluid in Ect1 cells, where TGFB, prostaglandins and TLR4 ligands all contribute to triggering the female response. Conclusions: Seminal fluid signaling elicits female tract cytokine and microRNA responses that cause (i) direct effects on the embryo’s developmental program, and (ii) indirect effects on implantation success and fetal development through the agency of regulatory T cells. The balance of different cytokines, prostaglandins and TLR ligands in seminal fluid explains substantial variation in the character of the female response elicited by individual seminal fluid samples. Thus we are gaining clarity on how seminal fluid can impact fertility and progression of pregnancy, as well as the female tract microbiome and infection status.

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Male-female seminal fluid communication – new insights SA Robertson, JE Schjenken, B Zhang, DJ Sharkey

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Role of seminal plasma in the transmission of HIV-1

Robinson Institute, School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, Australia

D Anderson, J Marathe, A Islam

Problem: Seminal fluid interacts with the female reproductive tract at coitus to induce a range of molecular and cellular changes that maximize the likelihood of reproductive success. We aim to define the signaling agents and to characterize the mechanisms of female tract response, including the role of microRNAs in transmitting and mediating this signaling cascade. Methods of Study: Mouse models including mice with null mutations in microRNAs have been utilized to define how sperm and seminal plasma interact with female tract cells. To investigate the human response to seminal fluid, in vitro experiments in Ect1 ectocervical cells are utilized. Results: In mice, both the plasma and sperm fraction of seminal fluid trigger gene expression changes in uterine and oviduct epithelial cells to elicit a cascade of cytokine and microRNA responses. These include upregulation of embryotrophic cytokines GM-CSF and LIF, and suppression of embryotoxic cytokine TRAIL. MicroRNAs including miR-223 induced in the female tract by seminal fluid alter the phenotype of dendritic cells and genetic ablation interferes with regulatory T cell induction and progression of pregnancy. Similar cytokine and microR-

Human seminal plasma contains a rich array of cytokines, chemokines and other immunological mediators that may play a role in the transmission of infectious pathogens. Normal human semen contains very high concentrations (>1 ng/ml) of TGF-b1, interleukin (Il)-7, SDF-1, MCP-1 and Il-8. Semen from men with inflammation resulting from genital infections can contain high concentrations of other proinflammatory cytokines and chemokines. The chemokines in human semen attract leukocytes into cervical tissue after intercourse and may enhance HIV transmission through the recruitment of HIV target cells. On the other hand, semen is toxic to T lymphocytes in high concentration, and has a concentration-dependent effect on HIV infection in vitro, showing suppression at high concentrations and enhancement at low concentrations in some studies. Macrophages appear to be resistant to the cytotoxic effects of semen and remain functionally intact after exposure to even high concentrations of seminal plasma. HIV-infected seminal macrophages are viable and capable of transmitting HIV across vaginal epithelia. Early studies using cervical epithelial cell monolayers indicated that the expression of a number of immunologically relevant genes, especially

Boston University School of Medicine, MA, USA

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proinflammatory cytokines and chemokines, are dramatically upregulated in epithelial cells following exposure to semen. However, our studies in a differentiated vaginal tissue model have shown that apical application of semen has little effect, possibly because the upper layers of cells in stratified epithelia are cornified and poorly able to respond to external stimuli. When the cornified barrier is damaged (i.e. by invasive coinfections or injury), seminal plasma may reach the metabolically active basal epithelial cells to further promote genital inflammation and HIV infection.

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Interplay between hormones, mucosal immunology and susceptibility to HIV infection in the human female reproductive tract CR Wira, M Rodriguez-Garcia, JV Fahey Department of Physiology and Neurobiology, Geisel School of Medicine at Dartmouth, NH, USA

Problem: Despite extensive studies of the mucosal immune system in the female reproductive tract (FRT) and its regulation by sex hormones, little attention has been paid to the tissue environment in the FRT that regulates immune cell function. Epithelial cells, fibroblasts and immune cells from the upper (Fallopian tubes, endometrium [EM], endocervix [CX]) and lower (ectocervix [ECX] and vagina) tracts contribute to immune protection. Each compartment is unique and precisely regulated by sex hormones to optimize conditions for pregnancy and protection against sexually transmitted infections (STI) including HIV. Methods of Study: Tissues from women undergoing hysterectomies were enzymatically digested and epithelial cells, fibroblasts, T cells and macrophages were isolated. Immune parameters including cytokine, chemokine and antimicrobial secretion, cell phenotype and function of cells in culture were analyzed for the direct and indirect effects of sex hormones (E2: estradiol and P4: progesterone). CD4+ T cell and macrophage susceptibility to infection was evaluated by HIV (BaL) infection. Results: Secretion of cytokines/chemokines, antimicrobials and growth factors by epithelial cells and fibroblasts are hormonally regulated and vary with the compartment analyzed. In some cases, secretion

is due to the direct effects of E2 and P4 while others are indirectly mediated through growth factors (TGFb, HGF, etc). Similarly, through hormonal regulation of the tissue environment, immune cell phenotype and function vary in a site specific way. For example, CD4+ Th17 cells, which represent a major subset in FRT tissues, are lower in the EM relative to the CX and ECX. CD8+ T cell and NK cell function is selectively suppressed at particular sites in the FRT. With regard to HIV susceptibility, EM epithelial cell and fibroblast secretions inhibit HIV infection of target cells. Moreover, E2 treatment prior to infection significantly reduces infection in CD4+ T cells and macrophages, the main target cells for HIV. Conclusions: This review examines the contributions of epithelial cells, fibroblasts and immune cells to the tissue environment in the FRT and the impact of this environment on susceptibility to HIV infection. While much remains to be learned about the complex interactions in tissues throughout the FRT, it is clear that sex hormones and chemical contraceptives regulate this environment to influence immune protection against HIV and other STI. Awareness of the involvement of the tissue environment in determining immune cell function and HIV susceptibility is crucial for understanding the mechanisms involved in HIV acquisition and will help develop strategies for HIV prevention. Supported by NIH contract HHSN272201000001C and NIH grants AI102838 and AI071761 (CRW).

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Hormonal contraception and HIV acquisition C Morrison Clinical Sciences, FHI 360, NC, USA

Hormonal contraception (HC) is used by >150 million women worldwide and the injectable progestin DMPA (depot medroxyprogesterone acetate) is the most widely used contraceptive in Sub-Saharan Africa. For >25 years, some epidemiologic studies have found that women using HC, particularly DMPA, have increased risks of HIV acquisition; however, study results are conflicting. Significant biologic data support a possible association between DMPA and HIV acquisition. Numerous macaque studies suggest that DMPA enhances simian immunodeficiency virus (SIV) acquisition. Bio-

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logic studies in humans suggest several mechanisms by which DMPA could increase HIV susceptibility. First, DMPA results in a hypoestrogenic state; loss of estrogen in menopause is associated with loss of vaginal acidity and protective lactobacilli, vaginal thinning and loss of antimicrobial secretion. MPA binds to most receptors with greater affinity than progesterone; in particular MPA’s high affinity for the glucocorticoid receptor may result in immunosuppression. Additional data support genital tract immunosuppression, e.g. DMPA is associated with changes in cervico-vaginal proteins (increased RANTES, decreased SLPI) predictive of HIV acquisition. Additional possible mechanisms include the effect of HC on vaginal structure (e.g. cervical ectopy), increased recruitment of HIV susceptible cells to the genital tract, increased risk of other STI and the direct effects of HC on HIV (e.g. up-regulation).

To date, 6 of 17 prospective studies of DMPA and 2 of 17 prospective studies of oral contraceptives have found increased risks of HIV acquisition; all have been observational. A 2012 WHO consultation advised no limitations on HC use among women at risk of HIV but stated that women using progestogen-only injectables be strongly recommended to always use condoms. WHO also recommended that higher quality clinical studies (i.e. randomized trials) and studies to understand the biology of the HC-HIV interaction are urgently needed. An RCT of injectable progestins and copper IUDs and HIV incidence is currently being planned.

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AJRI AWARD LECTURE

Developmental programming of the preimplantation embryo by colony stimulating factor 2 – a case of male versus female

S.04-1

Angiogenic biomarkers in preeclampsia S Ananth Karumanchi Howard Hughes Medical Institute, Beth Israel Deaconess Medical Center and Harvard Medical School, MA, USA

P Hansen Department of Animal Sciences, University of Florida, FL, USA

Colony stimulating factor 2 (CSF2) is a maternallyderived cytokine that acts during the preimplantation period to regulate embryonic development. In the cow, mouse, and human, CSF2 improves competence of embryos cultured in vitro to establish pregnancy after transfer to females. Actions of CSF2 on developmental competence involve inhibition of apoptosis and actions that increase the number of inner cell mass (ICM) cells. In the cow, CSF2 regulates genes involved in WNT signaling and differentiation in a manner that suggests CSF2 promotes pluripotency. Moreover, isolated ICM is more likely to survive passage in a pluripotent-state when derived from CSF2-treated blastocysts than when from control blastocysts. CSF2 can be considered as a developmental programming molecule because actions in the preimplantation period cause changes in the developmental program later in pregnancy. In cattle, for example, exposure of embryos to CSF2 from Day 5-7 of development (i.e., morula and blastocyst stages) results in embryos with altered capacity to undergo trophoblast elongation at Day 15 of gestation. The magnitude of this action of CSF2 depends on embryo sex. In male embryos, CSF2 treatment from Day 5 to 7 increases embryo length at Day 15 and increases secretion of the pregnancy recognition protein, interferon-s. In female embryos, CSF2 results in smaller embryos at Day 15 and reduced secretion of interferon-s. It is suggested that one way maternal environment can exert differential effects on male and female offspring is because maternal signaling molecules like CSF2 regulate development in a sex-dependent manner.

Imbalance of angiogenic growth factors in the maternal circulation contributes to the pathogenesis of preeclampsia. Soluble fms-like tyrosine kinase 1 (sFlt1), an endogenous anti-angiogenic protein that antagonizes vascular endothelial growth factor (VEGF) and placental growth factor (PlGF) appears to be a central player in this paradigm. Overexpression of sFlt1 in pregnant rats produced hypertension, proteinuria and glomerular endotheliosis, the classical pathological renal lesion of preeclampsia. High serum sFlt1 and low serum free PlGF and free VEGF have been observed in preeclampsia. Abnormalities in these circulating angiogenic proteins are not only present during clinical preeclampsia, but also antedate clinical symptoms by several weeks. Another potential soluble factor secreted by the placenta that appears to be elevated in women with preeclampsia is soluble endoglin (sEng). Endoglin (Eng) is an angiogenic receptor expressed mainly on the surface of endothelial cells, but also by placental syncytiotrophoblast. Eng acts as a co-receptor for transforming growth factor-beta, a potent pro-angiogenic molecule) signaling in endothelial cells. Eng mRNA is up-regulated in the preeclamptic placenta. In addition, the extra-cellular region of endoglin referred to as sEng, is released in excess quantities into the circulation of preeclamptic patients. Furthermore, sEng appeared to exacerbate the vascular damage mediated by sFlt1 in pregnant rats resulting in severe preeclampsia-like illness including the development of thrombocytopenia and fetal growth restriction. Measurement of angiogenic markers in the serum/plasma is particularly useful for the diagnosis and prediction of preterm preeclampsia. Methods aimed at interfering sFlt1 and/or sEng actions may be a novel therapeutic strategy in severe preterm preeclampsia.

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Development of an in-vivo equine herpesvirus-1 abortion model

The sweet kiss of galectin-1 (gal-1) during pregnancy: implications in preeclampsia

G Soboll Hussey1, SB Hussey1, DW Gardiner2, DP Lunn3, K Osterrieder4, Y-W Chiang5, P McCue2, F Del Piero6 1

PDI, Michigan State University, MI, USA; 2CVMBS, Colorado State University, CO, USA; 3North Carolina State University, NC, USA; 4Freie Universit€at Berlin, Germany; 5Fort Dodge Animal Health, USA; 6 Pathobiological Sciences, Louisiana State University, LA, USA

Equine herpesvirus-1 (EHV-1) continues to cause both sporadic and epidemic abortions primarily in the third trimester of pregnancy. Despite the significant impact of EHV-1 abortions, equine models employing current technology to characterize the outcomes of challenge infections are lacking. The objective of the current study was to develop an equine model of experimental EHV-1 abortion and use real time quantitative PCR to characterize viral infection of the placenta and fetus. Challenges with two different viral strains of EHV-1 were conducted in 7 pregnant pony mares at the 9th month of gestation. For each challenge, pregnancy was synchronized in ponies to one of two consecutive heat cycles by use of oral progestagens, hormonal therapy, and natural breeding. Prior to and subsequent to challenge infection all ponies were subject to clinical examination, nasal swab and blood leukocyte sampling for detection of EHV-1 infection, and sampling of any aborted fetus and placenta. All mares in both groups exhibited nasal viral shedding and viremia. Infection resulted in 1/7 (strain OH03) abortion and 5/7 abortions (strain Ab4) respectively. Aborted fetuses expressed high levels of virus infection in the spleen and liver, lower levels in the lung and thymus, and lowest levels in the chorioallantois. In the two normal deliveries of the Ab4 infected mares, the chorioallantois contained virus levels comparable with the chorioallantois of aborted foals and both foals shed EHV-1 starting on day 4 of life, but were clinically healthy. Our established in vivo model will facilitate the evaluation and development of vaccines for protection against EHV-1, as well as permit for studies aimed at understanding immunological processes and pathogenesis at the fetal/ maternal interface. Furthermore, our results demonstrate the continued importance of strain selection for EHV-1 abortion models.

SM Blois1, G Barrientos2, N Freitag1, M Conrad1  Center 12 Internal Medicine and Dermatology, Medicine Charite gica, University Berlin, Germany; 2Departamento de Quımica Biolo Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina 1

Problem: Although galectin-1 was identified more than 30 years ago, the role of this lectin in pregnancy is only emerging. Methods of Study: Different mouse models, including Lgals1 deficient mice, were used to investigate the role of gal-1 associated to normal pregnancy and pathological situations (e.g. preeclampsia. Results: Specific expression of galectin-1 has been identified at the maternal and placental compartments, where deregulated gal-1 functions have been associated with adverse pregnancy outcomes. Recent studies highlight the contribution of gal-1 to critical biological events occurring during mammalian gestation including immune cell tolerance, inflammation, implantation, and angiogenesis. Conclusions: Gal-1 has emerged as an important lectin with major functions during a physiological event like pregnancy.

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Inflammatory state of pancreatic islets in intrauterine growth restricted fetuses SW Limesand1,2, AC Kelly1,2, CA Bidwell1,2, FM McCarthy1,2 1 School of Animal and Comparative Biomedical Sciences, University of Arizona Department of Animal Sciences, AZ, USA; 2Purdue University, IN, USA

Problem: Intrauterine growth restriction (IUGR) has been associated with an increased incidence of Type 2 Diabetes Mellitus in adulthood. These studies indicate that developmental adaptions to conditions that result in IUGR permanently impair insulin-producing pancreatic b-cells. Growth restricted fetuses have decreased b-cell mass and reduced insulin secretion. A sheep model of placental insufficiency-induced IUGR has similar deficiencies in b-cells. Our objective was to measure gene expression of the islet

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exome from IUGR fetuses to elucidate mechanisms responsible for b-cell dysfunction. Method of Study: At 0.9 of gestation placental insufficiency-induced IUGR (established by environmental hyperthermia) and control sheep islets were isolated (n = 4/treatment). Isolated RNA was sequenced with Illumina hiseq2500 and analyzed using the Tuxedo Suite. Reference annotation based assembly using both bovine and ovine genomes identified differentially expressed transcripts. RNA transcripts were queried for enrichment and modeled to functional pathways previously defined. Results: In IUGR islets, 197 transcripts were upregulated and 493 were downregulated compared to control islets using the ovine genome reference. For the bovine genome reference 425 transcripts were upregulated and 652 downregulated. The two lists were combined for pathway analysis. For downregulated genes pathways over-represented were cytokinecytokine receptor interactions (4.8%) and chemokine signaling pathway (2.3%). IUGR islets had reduced expression of CCL2, CCL3, CCL4, CCL5, CCL21, CXCR4, and a portion of the interleukin family of cytokines including IL-1b, IL-8, and IL-6. IUGR islets expressed greater concentration of CXCL12, IL-6R and GPR75. Conclusions: Placental insufficiency-induced IUGR fetuses have decreased pro-inflammatory chemokines and cytokines that may regulate homeostatic processes in the islets. Previous findings show that IUGR suppresses the inflammatory response in postnatal animals and transiently alters neonatal islet T-helper cells in islets. However, our findings also indicate a role for cytokines in b-cell dysfunction because the CXCL12/CXCR4 axis promotes pancreatic b-cell growth and genesis in human and rodent systems.

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The role of TLR signaling in fetal brain injury from prenatal inflammation M Elovitz Department of Obstetrics and Gynecology, University of Pennsyvlania Perelman School of Medicine, PA, USA

Infection and inflammation remain a significant contributor to pregnancy loss and infant morbidity and mortality. Maternal infections during pregnancy represent a significant risk factor for adverse neurobehavioral outcomes, including cognitive delay,

schizophrenia, autism and mental retardation. While antenatal infections are associated with these adverse outcomes, whether the pathogen itself or pathogen by-products are responsible for activating an immune response and subsequent fetal injury is unclear. Common organisms that infect pregnant women include E. coli, Group B streptococcus (GBS), Mycoplasma and Ureaplamsa; all of these pathogens are implicated in perinatal injury and exposure to these organisms has been associated with long term consequences for the offspring. These particular organisms are most likely to contribute to perinatal morbidity through colonization and/or infection in the uterine cavity but not from systemic infection of the mother. Understanding this more common clinical scenario, we have developed mouse models that accurately mimic intrauterine infection and inflammation in the human. Research from our laboratory has demonstrated that intrauterine inflammation results in both fetal and postnatal brain injury. Recently, we have demonstrated that even with a low dose model of intrauterine inflammation-in which there is minimal immune response in the uterus-the presence of bacterial by-products in the uterine cavity can cause fetal and postnatal brain injury. While activation of the immune response in both the mother and fetoplacental unit is believed to play a crucial role in the pathogenesis of fetal brain injury from prenatal infection, the necessity of Toll-like receptor (TLR) signaling pathways in the mother, placenta and/or fetus to inflammation-induced fetal brain injury has not been elucidated. Until the immune pathways mechanistically responsible for adverse outcomes from prenatal infection are revealed, therapeutic strategies cannot begin. Targeting maternal TLR pathways may be an important therapeutic strategy to prevent adverse outcomes; however, this approach may be for naught if pathogen or pathogen by-products can directly activate fetal TLR in the absence of a maternal immune response and TLR therapies do not reach the fetal compartment. Moreover, if immune mediators activate non-TLR pathways, in either the mother or fetus, targeting TLR signaling would fail to prevent outcomes. Using our existing mouse models, genetically manipulated mice and embryo transfer techniques, we have begun to dissect the involvement of TLR signaling in adverse outcomes from prenatal inflammation.

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Maternal nutritional status, genital tract inflammation, and preterm birth H Simhan University of Pittsburg, School of Medicine, PA, USA

In this presentation, we will discuss the relation between maternal nutritional status during pregnancy and preterm birth. We will discuss several aspects of maternal nutritional status, including body composition, folate status, and vitamin D status. We will also delve into possible mechanisms connecting maternal nutritional status and preterm birth, with a focus on genital tract flora and immunological milieu. With respect to body composition, we will discuss the relation of both pre-pregnancy body mass index

(BMI) and gestational weight gain (GWG) on preterm birth. We will dissect the contribution of BMI and GWG to spontaneous and indicated preterm birth, and review the basis for clinical recommendations. With respect to folate, we will discuss the differential and interactive contribution of the dominant folate vitamers to the risk of preterm birth, and how this biology may work through the lower genital tract inflammatory milieu to predispose to ascending microbial invasion of the upper genital tract during pregnancy. With respect to vitamin D, we will discuss the influence of vitamin D on length of gestation based on timing of exposure, and address the contribution of vitamin D to lower genital tract flora.

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J. CHRISTIAN HERR LECTURE

Novel mechanisms of placental inflammation in obstetric antiphospholipid syndrome

and, thus, provide new insights into our understanding of the pathogenesis of pregnancy complications in women with APS.

VM Abrahams Department of Obstetrics, Gynecology & Reproductive Sciences, Yale School of Medicine, CT, USA

Antiphospholipid syndrome (APS) is an autoimmune disorder, which places pregnant women at increased risk for recurrent pregnancy loss (RPL) and late gestational complications, such as preeclampsia. While treatment with heparin reduces the risk for RPL, the incidence of severe complications, like preeclampsia, remains high in these patients. Thus, there remains a need to better understand the pathophysiology of obstetric APS, so that we can better to identify novel therapeutic approaches. APS is characterized by circulating antiphospholipid antibodies (aPL) that bind beta 2 glycoprotein I (b2GPI) expressed by the trophoblast, leading to altered placental function. Studies from our lab have shown that aPL directly affect human first trimester trophoblast cells by: (i) inducing a pro-inflammatory cytokine/chemokine response; (ii) disrupting the normal angiogenic factor profile; and (iii) limiting trophoblast migration and interactions with the endothelium. Furthermore, we have investigated the mechanisms by which aPL trigger the trophoblast to generate an inflammatory cytokine/chemokine response at the molecular level. We have found that aPL elevate trophoblast IL-1b and IL-8 production in a TLR4/MyD88-dependent manner. However, downstream of TLR4, the aPLinduced signaling pathways triggering these inflammatory factors diverge. aPL through TLR4 induce trophoblast cells to produce their own uric acid, which in turn activates the Nalp3/ASC inflammasome, leading to trophoblast IL-1b processing and secretion. In parallel, we describe for the first time that aPL through TLR4 induce the trophoblast to express high levels of certain microRNAs, which instead of acting as a negative regulators of inflammation, though their release in exosomes, promote aPL-induced trophoblast IL-8 secretion. Together our studies indicate novel cellular and molecular mechanisms by which aPL trigger placental inflammation

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The perinatal microbiome: how does pregnancy shape our microbiome, and how does our microbiome shape us? K Aagaard Obstetrics & Gynecology, Division of Maternal-Fetal Medicine at Baylor College of Medicine, TX, USA

Hominids and hominins serve as remarkable hosts to microbes, and we have co-evolved over the past 4.5 million years as highly plethoric communities. Precisely when and how these microbes take up residence during development and over the span of an individual’s lifetime remains unclear. The burden of perinatal (pregnancy and newborn) morbidity and mortality related to infection and/or inflammation is astounding. In 2005, the WHO estimated that 9.6% or 12.9 million births worldwide were born preterm at

Abstracts of the 34th Annual Meeting of the American Society for Reproductive Immunology, 2-5 june, 2014, Long Beach, New York.

Abstracts of the 34th Annual Meeting of the American Society for Reproductive Immunology, 2-5 june, 2014, Long Beach, New York. - PDF Download Free
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