bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Age on Reproductive Outcome in Women With Recurrent Spontaneous Abortions and Infertility of Immune Etiology
Authors: Ruiz A.M. , Kwak J.Y.H. , Kwak F.M. , Beer A.E.
Published: September 2011

The objective of this paper is to determine whether age has any impact on conception rate, pregnancy outcome, or autoimmune status in women with recurrent spontaneous abortions (RSA) and infertility of immune etiology. One hundred twenty‐four women with 3 or more RSA and 36 women with unexplained infertility were prospectively studied. Maternal anti‐paternal lymphocyte antibodies and autoantibodies to phospholipids and nuclear antigens were tested. All achieved an adequate alloimmune recognition after lymphocyte immunization and followed for 1 year with optimal preconception autoimmune treatment. Conception rate and pregnancy outcome were prospectively studied. 1) 10.1% of women with RSA and 30.6% of women with infertility failed to achieve a pregnancy after 1 year of trial (P=0.0084); 2) in women with RSA, the number of previous fetal death after 28 weeks of gestation was significantly higher in women who failed to achieve a pregnancy within 1 year when compared to women who became pregnant (P=0.0296). Conception rate was not different with advancing age; 3) women with infertility demonstrated significantly higher incidence of anti‐phosphatidylethanolamine antibody when compared to women with RSA (P=0.052); 4) in women with infertility, those who failed to achieve a pregnancy were significantly older (P=0.0012) and demonstrated a higher incidence of autoantibodies to phosphatidic acid than women with infertility who became pregnant (P=0.0339); 5) the subsequent spontaneous abortion rate while on optimal immune therapy was the same in the women with RSA (39.3%) and women with infertility (32%). Spontaneous abortion rate of women with RSA or infertility was not different among four age groups; 6) the presence of anti‐cardiolipin antibody (P=0.0055) and higher gravidity (P=0.0354) correlated significantly with pregnancy failure in women with a history of infertility. The prevalence of auto‐antibodies to phospholipids and nuclear components was not different among four age groups in women with RSA or infertility. Age does not affect pregnancy outcome or conception rate in women with RSA. In women with infertility of immune etiology, conception rate was significantly reduced over age 40 although pregnancy outcome was no different with advanced age.
Mucosal Immunity in the Human Female Reproductive Tract: Cytotoxic T Lymphocyte Function in the Cervix and Vagina of Premenopausal and Postmenopausal Women
Authors: White Hillary D. , Yeaman Grant R. , Givan Alice L. , Wira Charles R.
Published: September 2011

PROBLEM: To investigate the mucosal immune system in the cervix and vagina of premenopausal women in terms of immune cells present and cytolytic capacity of mucosal CD3+ T cells in the lower reproductive tract.

METHODS: Fresh tissue fragments prepared by vibratome sectioning were analyzed for the presence of immune cells by confocal scanning laser microscopy (CSLM). Isolated reproductive tract cells prepared by enzymatic digestion were analyzed for CD3+ T cell phenotype by FACS analysis and for cytolytic function by an anti‐CD3 mAb mediated redirected lysis assay.

RESULTS: As determined by CSLM, CD3+ cells as well as macrophages and dendritic cells are distributed throughout the lower female reproductive tract in both the epithelium and subepithelial mucosa. It was found that cervical and vaginal tissues from pre‐ and post‐menopausal women contain CD3+ T cells (CTL) that have cytolytic activity, when measured in an antigen non‐specific anti‐CD3 mAb mediated redirected lysis assay.

CONCLUSIONS: These results indicate that the lower reproductive tract of women is immuno‐competent as judged by the presence of CD3, CD4, CD8, macrophage, and dendritic cells in the endocervix, ectocervix, and vagina of premenopausal and postmenopausal women. Further, these studies demonstrate that CD3+ T cells with cytolytic activity are present in the cervix and vagina during the proliferative and secretory phases of the menstrual cycle and following menopause.
Sperm Antigens and Their Use in the Development of an Immunocontraceptive
Authors: Diekman Alan B. , Herr John C.
Published: September 2011

PROBLEM: Overpopulation is a global problem of significant magnitude, with grave implications for the future. World population is predicted to reach 10 billion people by the year 2050, an increase of more than 75% over the current population. Development of new contraceptives is necessary, since current forms of birth control are unavailable to many individuals due to sociological, financial, or educational limitations.

METHODS: In an effort to identify new contraceptive alternatives, sperm‐specific antigens are under investigation as the basis for immunological regulation of fertility through contraceptive immunization. Anti‐sperm antibodies (ASA) inhibit sperm function in vitro, induce infertility in experimental models, and have been implicated in some cases of clinical infertility. Furthermore, the development of ASA in vasectomized men does not lead to physiological complications despite the persistence of these antibodies for years; thus, ASA induced by immunization of men and women may be similarly harmless. Immunization has several conceptual advantages as a method of fertility regulation: Employment of non‐pharmacologically active agents, convenience of administration, low cost, and the potential for relatively long‐lasting yet reversible effects.

RESULTS: Various approaches have been taken to identify candidate sperm antigens for immunocontraceptive development. Studies that utilized monoclonal antibodies and polyclonal antisera generated against sperm preparations have identified such promising candidates as PH‐20, fertilin, SP‐17, and SP‐10. In an animal model, 100% contraceptive effects were induced following PH‐20 immunization and the effects were reversible. More recently, ASA from the sera of infertile patients were employed to identify several candidates as expressed by testis cDNA libraries. Perhaps the most extensively characterized and effective immunogen candidate identified thus far is lactate dehydrogenase‐C4 (LDH‐C4). Active immunization with LDH‐C4 suppressed fertility in a variety of mammalian species, including primates, and the reversibility of these effects was demonstrated.

CONCLUSION: The successful results obtained thus far support the feasibility of an effective immunocontraceptive and indicate the importance for continued investigation of additional sperm antigens as contraceptive immunogen candidates.
The HSD‐hCG Vaccine Prevents Pregnancy in Women: Feasibility Study of a Reversible Safe Contraceptive Vaccine
Authors: Talwar G.P. , Singh Om , Gupta S.K. , Hasnain S.E. , Pal R. , Majumdar S.S. , Vrati S. , Mukhopadhay A. , Srinivasan J. , Deshmukh U. , Ganga S. , Mandokhot A. , Gupta A.
Published: September 2011

PROBLEM: To develop a vaccine for reversible control of fertility in women.

MATERIALS AND PROTOCOLS: Purified β subunit of hCG annealed to purified alpha subunit of ovine LH linked chemically to tetanus toxoid (TT) and diphtheria (DT); vaccine employed at 300 μg gonadotropin equivalent per injection adsorbed on alhydrogel with 1 mg SPLPS added in the first injection; Phase I safety trials in 47 women with elective tubal ligation; Phase II efficacy studies in 148 proven fertile women (2 children), sexually active, desirous of family planning using IUD; IUD removed when anti‐hCG titres exceed 50 ng/ml hCG bioneutralization capacity; boosters given to maintain above threshold antibody levels; post coital tests conducted in 8 volunteers; sera of protected women analysed for immuno‐determinants recognized by competitive enzyme immunoassays employing a panel of monoclonal antibodies and by direct binding to synthetic peptides; recombinant vaccines expressing phCG as a secreted product or as a fused protein anchored on membrane.

RESULTS: Immunization was well tolerated with no significant changes in endocrine, metabolic and hematological indices. Normal ovulatory cycles were maintained as indicated by menstrual regulation. The vaccine was highly effective in preventing pregnancy (1 pregnancy in 1224 cycles) at and above antibody titres of 50 ng/ml. Antibodies declined in course of time in absence of boosters, with conceptions occurring below 35 ng/ml titres indicating regain of fertility. Ability of antibodies to prevent pregnancy was confirmed by post coital tests. High avidity (1010M—1) and other characteristics of antibodies generated by the vaccine are described and compared with those induced by two other hCG vaccines having undergone Phase I trials. The antibody response of the HSD vaccine in humans is characterized predominantly to an epitope recognized by the monoclonals 206 and P3W80. The antibodies had low or no reactivity with the carboxy terminal peptide and 38–57 region peptide. Live recombinant vaccines expressing βhCG as a membrane anchored peptide generated antibody response to hCG in all animals following a single injection.

CONCLUSIONS: Reversible fertility control is feasible with the HSD‐hCG vaccine without impairment of ovulation or disturbance of menstrual regularity. Suggestions have been made for further optimization of the vaccine, which include replacement of TT and DT by a panel of T non B determinants communicating with the entire MHC spectrum and development of recombinant vaccine expressing phCG along with membrane anchored carrier.
The Presence of Antithyroid Antibodies in Euthyroid Patients With Unexplained Infertility and Tubal Obstruction
Authors: Geva Eli , Lessing Joseph B. , Lerner‐Geva Liat , Azem Foad , Yovel Israel , Amit Ami
Published: September 2011

PROBLEM: The presence of antithyroid antibodies in euthyroid patients with unexplained infertility and tubal obstruction.

METHOD: The presence of antithyroid autoantibodies (microsomal and thyroglobulin) was measured in 40 patients with unexplained infertility, and 40 patients with tubal obstruction infertility, and compared to 40 healthy nulligravidae.

RESULTS: Eight patients (20%) in the unexplained infertility study group, seven (17.5%) in the tubal obstruction group and two (5%) in the healthy nulligravida group, were positive for antithyroid autoantibodies: five (12.5%) were positive for antimicrosomal antibodies, two (5%) were positive for antithyroglobulin antibodies, and one patient (2.5%) was positive for both. The tubal obstruction group comprised seven (17.5%) patients positive for antithyroid autoantibodies: four (10%) for antimicrosomal antibodies, two (5%) for antithyroglobulin antibodies, and one patient (2.5%) was positive for both. In the healthy nulligravidae group only two patients (5%) were positive for antithyroid antibodies: one for antimicrosomal and one for antithyroglobulin. No significant differences were found in the presence of antithyroid antibodies between patients with unexplained infertility and those with tubal obstruction infertility. Both groups differed significantly from the healthy controls with regard to the presence of antithyroid antibodies (P < 0.05).

CONCLUSION: Subclinical presence of antithyroid autoantibodies is characteristic of both unexplained and mechanical infertility, as opposed to healthy controls. Further investigation of larger groups is needed to determine the prevalence of antithyroid antibodies in the unique population of infertile women.
Placental Cytokines and the Pathogenesis of Preeclampsia
Authors: CONRAD KIRK P. , BENYO DEBORAH F.
Published: September 2011

The authors explore the hypothesis that tumor necrosis factor‐α (TNF‐α) and possibly other inflammatory cytokines are overproduced by the placenta in response to local ischemia/hypoxia contributing to increased plasma levels, and subsequent endothelial activation and dysfunction in the pregnancy disorder, preeclampsia. It is widely held that inadequate trophoblast invasion and physiologic remodeling of spiral arteries initiate placental ischemia/hypoxia in preeclampsia. Furthermore, focal areas of placental hypoxia have been implicated in the production of “toxic” factor(s) by the placenta, which circulate and cause maternal disease. Placental trophoblast cells and fetoplacental macrophages normally produce TNF‐α and in‐terleukin‐1 (IL‐1), which are capable of producing endothelial cell activation and dysfunction. Hypoxia has recently been reported to increase TNF‐α and IL‐1 production by term villous explants from the human placenta. Placental cells also express erythropoietin (EPO), which is the prototype molecule for transcriptional regulation by hypoxia in mammals. Interestingly, TNF‐α and IL‐1 have DNA sequences homologous or nearly homologous to the hypoxia‐responsive enhancer element of the EPO gene, thus providing a potential, but as of yet, untested molecular link between placental hypoxia and stimulation of cytokine production. Inflammatory cytokines overproduced by the placenta in response to hypoxia may then lead to increased plasma levels and endothelial activation and dysfunction in preeclampsia. The purpose of this short review is to critically evaluate the hypothesis that placental cytokines contribute to the pathogenesis of preeclampsia. Of note, the etiology of the disease presumably related to deficient trophoblast invasion is beyond the scope of this work.
Effects of Serum from Pregnant Versus Nonpregnant Women on Natural Killer Cell Activity
Authors: Fujino Yuji , Nakamura Yoshihiro , Ueda Kayoko , Ozaki Kouji , Ito Fumitoshi , Sun Ting‐Ting , Umesaki Naohiko , Ogita Sachio
Published: September 2011

PROBLEM: To test the effect of serum obtained from in vitro fertilization‐embryo transfer (IVF‐ET) patients on the healthy volunteers natural killer (NK) cell activity.

METHOD OF STUDY: Retrospective non‐randomized clinical study. Suppressive effect on NK cell activity was measured with serum obtained from 25 pregnant and 24 non‐pregnant women during IVF‐ET procedure.

RESULTS: Suppressive serum effect of volunteers' NK cell activity was significantly higher in pregnant than in non‐pregnant women (P < 0.01).

CONCLUSIONS: The suppressive activity of serum from pregnant women on NK cell suppression activity was useful in predicting the establishment of a successful pregnancy.
Lactation Stage‐Dependent Changes of Lymphocyte Subpopulations in Mammary Secretions: Inversion of CD4+/CD8+ T Cell Ratios at Parturition
Authors: Yang T.J. , Ayoub Ihsan A. , Rewinski Michael J.
Published: September 2011

PROBLEM: Determination of lactation stage‐dependent changes in levels of lymphocyte subpopulations in milk.

METHOD: Flow cytometric assay was used to identify and assay lymphocyte subpopulations in bovine milk at different stages of lactation.

RESULTS: Lymphocyte subpopulations in mammary secretions of dairy cows change during the lactation cycle. In involuting glands (dry gland), ∼ 80–90% of lymphocytes were CD2+ T cells. The proportion of CD2+ T cells, however, decreased to ∼ 50% at the colostral stage and fluctuated between 50 to 60% in normal (mature) milk. Throughout the lactation stages, less than 5% were B cells as identified by the monoclonal antibodies against CD21 and MHC class II antigens. Subset analysis showed, however, that the proportion of CD5+ T cells decreased from 90% in involuting gland secretions to 75% in colostrum (peripartum stage), and to ∼ 40–50% in the normal (mature) milk. CD4+ T cells constituted between 45 to 55% of lymphocytes in the dry gland secretion but decreased drastically at parturition and maintained at the level below 20% throughout normal lactation. In contrast, the proportion of CD8+ T cells in the dry gland secretion was low, between 30 to 40%, but increased steadily, in an inversely‐related manner with that of CD4+ T cells, to ∼ 40–50% at parturition and maintained at ∼ 30–40% during the normal lactation stage thereafter. Two‐color immunofluorescence study revealed further that practically all of the CD8+ cells in dry gland secretions were CD2+, and approximately 40% of them were CD5—. Throughout the lactation cycle, WC1+γδ T cells comprised only 2 to 5% of lymphocytes in mammary secretions.

CONCLUSIONS: T lymphocyte subpopulations change dynamically during stages of the lactation cycle. The selective migration of T lymphocyte subpopulations to and from the mammary gland, and their functional roles in the immune competence and regulation of the dam and sucklings remain to be elucidated.
Estrogen Inhibits Fetal Thymocyte Development In Vitro
Authors: Rijhsinghani Asha , Bhatia Sudershan K. , Kantamneni Lakshmi , Schlueter Annette , Waldschmidt Thomas J.
Published: September 2011

PROBLEM: Transient involution of the maternal thymus in mice is known to occur during pregnancy. We have previously reported that the hormone responsible for this involution is estrogen. Interestingly, although estrogen crosses the placenta, fetal thymus gland enlarges with advancing gestational age. It is not known if fetal thymocytes are resistant to estrogen or if there are other factors that prevent estrogen from exerting an effect on the development of fetal thymocytes. Therefore we studied the effect of estrogen on isolated fetal thymic glands in vitro.

METHOD OF STUDY: Pregnant Balb/c mice were sacrificed at 15 days gestation and fetal thymic lobes were obtained from all fetuses. The glands were cultured in vitro using either control medium or medium to which estrogen was added in two concentrations of 0.5 mg/ 100 ml and 1.0 mg/100 ml. After 12 days of organ culture, total thymocyte counts and phenotypic analysis by three color flow cytometry were performed by using monoclonal antibodies to surface markers of T cell subsets.

RESULTS: Estrogen treatment caused a marked suppression of the total number of fetal thymocytes. All CD4 and CD8 defined T cell subsets were reduced with a disproportionate loss of CD4+ single positive (SP), CD8+ SP; CD4+CD8+ double positive (DP) cells. The early thymocyte developmental stages, based on CD44 and CD25 expression, revealed the CD4‐CD8‐CD3‐ triple negative compartment (TN) to be composed of almost entirely the earliest population (CD44+CD25‐) with the remaining maturational stages depleted.

CONCLUSIONS: This study demonstrates that fetal thymus removed from the intact fetus is susceptible to the inhibitory effects of estrogen. Since the fetal thymus enlarges with advanced gestational age, it is clear that the intact fetus invokes a regulatory mechanism which neutralizes the anti‐lymphopoietic action of estrogen observed in the adult female.
Interferon Tau: A Novel Pregnancy Recognition Signal
Authors: Bazer Fuller W. , Spencer Thomas E. , Ott Troy L.
Published: September 2011

PROBLEM: Trophectoderm of ruminant conceptuses (embryo and associated membranes) secretes tau interferons (IFNẗ) as the pregnancy recognition signal. How does it act?

METHOD: Review of current data.

RESULTS: IFNt acts on uterine epithelium to suppress transcription of the genes for estrogen receptor and oxytocin receptor. This blocks development of the uterine luteolytic mechanism and, therefore, release of luteolytic pulses of prostaglandin F2α, but it has no effect on expression of the progesterone receptor. Maintenance of progesterone secretion by the corpus luteum ensures establishment and maintenance of pregnancy. Secretion of IFNẗ on days 12–15 for sheep and days 14–17 for cows and goats is essential for pregnancy recognition.

CONCLUSION: We propose that IFNẗ affects endometrial gene expression by activating the Jak/Stat pathway, which results in formation of the ISGF3α transcription factor complex. ISGF3α binds to interferon‐stimulated response elements and activates transcription of interferon‐responsive genes such as interferon regulatory factor‐1 (IRF‐1) which, in turn, activates expression of the negative‐acting transcription factor IRF‐2. Pregnancy (or intrauterine injection of roIFNẗ) results in a transient increase in endometrial IRF‐1 expression followed 36–48 hr later by a sustained increase in IRF‐2. We propose that IRF‐2, or an IFNẗ‐induced negative regulatory factor like IRF‐2, suppresses expression of the estrogen receptor gene and directly or indirectly blocks expression of the gene for oxytocin receptor to abrogate the uterine luteolytic mechanism and ensure the establishment of pregnancy.
Immune Suppression and Th1/Th2 Balance in Pregnancy Revisited: A (Very) Personal Tribute to Tom Wegmann
Authors: Chaouat Gérard , Diallo Juliette Tranchot , Volumenie Jean Luc , Menu Elisabeth , Gras Gabriel , Delage Geneviéve , Mognetti Barbara
Published: September 2011

PROBLEM: The paradigm of local suppression necessary to understand the survival of the fetal allograft is often compared with the host‐tumor relationship.

METHODS: We investigated two components of local immune suppression: placenta‐induced immunosuppression, which is mediated at least in part by a soluble factor of low molecular weight that can induce anergy in lymphocytes, and interleukin‐10 (IL‐10).

RESULTS: We show that enhancement of IL‐10 production in the decidua and placenta after alloimmunization requires the presence of Asialo GM1+ cells. Placenta‐induced immunosuppression is linked with defects in phosphorylation of some components of the T cell receptor.

CONCLUSION: NK cells could be in fact regulatory cells pushing maternal immune response toward a Th2 profile, beneficial for fetal survival, or toward a Th1 type of immune response, which acts in synergy. Modulation of TcR may represent a new mechanism for maternal‐fetal tolerance.
Uterine NK Cells and Trophoblast HLA Class I Molecules
Authors: King A. , Hiby S.E. , Verma S. , Burrows T. , Gardner L. , Loke Y. W.
Published: September 2011

PROBLEM: To investigate the proposal that NK cells in decidua may control trophoblast migration during implantation of the human placenta.

METHOD: Use Mab specific for HLA‐G and for HLA‐C in association with flow cytometry and immunoprecipitation to determine the expression of these HLA molecules by trophoblast. Expression of Killer inhibitory/activatory receptors (KIR/KAR) and the CD94 receptor by decidual NK cells was also studied.

RESULTS: Extravillous trophoblast expressed HLA‐G and HLA‐C in both β2m‐associated form and as free heavy chains. KIR and KAR are expressed by decidual NK cells. The repertoire of receptors varied between different women and also between blood and decidual NK cells from the same women. The expression of CD94 was also different between blood and decidual NK cells.

CONCLUSION: The recognition of HLA‐G/HLA‐C by KIR/KAR and CD94 could provide a mechansm by which decidual NK cells control trophoblast migration.
Uterine Natural Killer Cells Do Not Require Interleukin‐2 for Their Differentiation or Maturation
Authors: Croy B. Anne , Guimond Marie‐Josee , Luross Jeffrey , Hahnel Ann , Wang Baoping , van den Heuvel Marianne
Published: September 2011

PROBLEM: Natural Killer lymphocytes (NK cells) from the pregnant uterus and from other tissues in pregnant and nonpregnant mammals can be stimulated by interleukin‐2 (IL‐2) during culture to become Lymphokine Activated Killer (LAK) cells. The susceptibility of cultured trophoblast cells to lysis by LAK cells raises the enigma of why uterine (u) NK cells that are characterized by morphology and by surface phenotyping as “activated,” and thus potentially damaging to the placenta, become localized to implantation sites during normal rodent gestation.

METHOD: uNK cells migrating from explant cultures of the metrial gland were assessed for expression (mRNA and protein) of each chain of the IL‐2 receptor (IL‐2R). Implantation sites from transgenic mice lacking a functional IL‐2 gene were examined histologically for the differentiation of mature, granulated uNK cells.

RESULTS AND CONCLUSION: Early post‐implantation, mRNA from migrating uNK cells contains transcripts for all three chains of the IL‐2R. Only IL‐2Rγ was expressed at day 12 of gestation; expression of this gene was also lost by day 16. Loss of IL‐2R transcription did not result in loss of protein expression; however, it did coincide with loss of uNK cell viability in vivo. Apparently normal differentiation of uNK cells occurred in IL‐2—/‐ mice and in doubly mutant IL‐2—/‐.β2m—/‐mice. Thus, despite uNK cell expression of the full 1L‐2R at day 8 of gestation, IL‐2 is not required for the maturation of uNK cells to their fully granulated form or for normal placental development.
Decidual Infiltration and Activation of Macrophages Leads to Early Embryo Loss
Authors: Baines M.G. , Duclos A.J. , Antecka E. , Haddad E.K.
Published: September 2011

PROBLEM: There is considerable controversy concerning the root cause and mechanisms of early embryo loss. It has been suggested that most pregnancy losses occur due to morphogenetic anomalies of the embryo. It has also been suggested that the maternal specific immune system rejects the embryo.

METHODS: Existing data on the cell and molecular biology of early embryo loss in murine experimental models is reviewed.

RESULTS: Using the CBA ♀ × DBA/2 ♂ model of early embryo loss, it has been established that maternal inflammatory cells infiltrate the decidua basalis of all implantation sites within 48 hr after implantation. For most embryos, the relatively low numbers of macrophages (MΦ) and natural killer‐like (NK‐like) cells of maternal origin remain relatively constant after day 8, whereas 20–30% of the embryos show a significant increase in inflammatory cells in the maternal decidua, corresponding to the incidence of early embryo resorption visible at day 12. Evidence will be reviewed to suggest that decidual NK‐like cells are not cytolytic but may be producing the MΦ‐activating cytokine interferon gamma (IFN‐γ), which activates decidual MΦ and other cells. Furthermore, embryo loss is ameliorated by in vivo treatment with anti‐IFNγ or anti‐NK antisera, indicating that NK‐like cells and/or IFNγ are required for embryo loss, but not for embryo survival. In resorbing embryos, the inflammatory MΦ show evidence of having been primed during early pregnancy, in that in vitro incubation with lipopolysaccharide induced the production of tumor necrosis factor alpha and nitric oxide.

CONCLUSION: These findings support the concept that early embryo loss is a nonspecific event mediated by the triggering of cytotoxin production by primed decidual macrophages.
Maternal Anti‐Placental Cell‐Mediated Reactivity and Spontaneous Abortions
Authors: Raghupathy Raj
Published: September 2011

PROBLEM: Does Th1/Th2 balance determine pregnancy outcome, and if so, what determines Th1/Th2 balance in pregnancy?

METHOD: Review and synthesis of existing data.

RESULTS: A bias toward Th1 is strongly correlated with pregnancy failure in mice and humans. Pregnancy usually shifts the balance toward Th2 and placental factors/progesterone and progesterone‐stimulated CD8+ T cell production of suppressor factor; TGFβ2 and IL4/ 10 may be responsible. The bias toward a Th1 response may result from intracellular parasitic infection and other as yet undefined factors.

CONCLUSION: The Th1/Th2 balance thesis appears to be valid.
Murine T Cell Determination of Pregnancy Outcome: I. Effects of Strain, αβ T Cell Receptor, γδ T Cell Receptor, and γδ T Cell Subsets
Authors: Arck Petra C. , Ferrick David A. , Steele‐Norwood Darlene , Croitoru Ken , Clark David A.
Published: September 2011

PROBLEM: T cells bearing αβ T cell receptor (TcR) and γδ TcR are present at the fetomaternal interface, and the latter, which express surface activation markers, can react with fetal trophoblast cell antigens. What is the role of these cells?

METHOD: Using stress‐abortion‐prone DBA/2‐mated CBA/J and abortion‐resistant C57/B16 mice, αβ, γδ, and CD8+/‐ T cell subsets were measured in spleen and uterine decidua. The effect of immunization against abortion and administration of anti‐TcR antibody in vivo was examined. Cytokine synthesis was measured by intracellular staining of Brefeldin A‐treated cells.

RESULTS: Abortion‐prone matings showed an unexpected accumulation of γδ T cells beginning in the peri‐implantation period and this was suppressed by immunization against abortion. The immunization deleted γδ T cells producing the abortogenic cytokines, TNF‐α and γ‐interferon, and increased production of the anti‐abortive cytokines, IL‐10 and transforming growth factor‐β2 (TGF‐β2). Immunization also boosted the number of αβ T cells which were present in the decidua as early as 2 days after implantation. In vivo injection of GL4 (anti‐δ) depleted γδ T cells producing Th1 cytokines in the peri‐implantation period, and prevented abortions, whereas H57 (anti‐β) decreased the number of αβ T cells and led to 100% abortions. CD8+ T cells present in peri‐implant decidua before onset of abortions were mostly αβ TcR+, although some were γδ+. Changes in γδ and αβ T cells in pregnancy were most dramatic in uterine tissue.

CONCLUSION: Although decidual γδ T cells after formation of a distinct placenta and fetus produce anti‐abortive TGF‐β2‐like molecules and IL‐10, prior events can lead to abortion. High local production of TNF‐α and γ‐interferon develop during the peri‐implantation phase because of an excessive increase in the Th1 cytokine+ subset of γδ cells; these cytokines may be contributed by other tissues in decidua, and the contribution of bioactive factors by γδ T cells may augment the cytokine pool. In contrast, αβ T cells (which may be inactivated by stress that causes abortions) may mediate the anti‐abortive effect of alloimmunization. Alloimmunization involves a shift from a Th1 to a Th2 pattern in the γδ T cells in decidua.
Presence of HLA‐G‐Expressing Cells Modulates the Ability of Peripheral Blood Mononuclear Cells to Release Cytokines
Authors: Maejima Masamoto , Fujii Tomoyuki , Kozuma Shiro , Okai Takashi , Shibata Yoichi , Taketani Yuji
Published: September 2011

PROBLEM: Human leukocyte antigen‐G (HLA‐G) is thought to be at play in maternal‐fetal immune interplay during pregnancy. Whether the expression of HLA‐G protein on the target cells altered the release of cytokines from effector mononuclear cells was questioned. METHOD OF STUDY: The amounts of cytokines released from peripheral blood mononuclear cells (PBMC) cocultured with or without HLA‐G‐expressing target cells were compared. RESULTS: When cocultured with HLA‐G‐expressing target cell lines, the amounts of interleukin‐3 (IL‐3) and interleukin‐lβ (IL‐1β) released from PBMC were increased, whereas the amounts of tumor necrosis factor‐α (TNF‐α) were decreased.

CONCLUSIONS: Mononuclear cells, if cultured with HLA‐G‐expressing cells, modulate their ability to release cytokines, suggesting a role of HLA‐G in triggering maternal‐fetal immune interplay and thereby maintaining pregnancy.
Effect of Boar Seminal Immunosuppressive Component on Humoral Immune Response in Mice
Authors: Veselský Leopold , ŽElezná Blanka , Dostál Jaromír
Published: September 2011

PROBLEM: The effect of seminal immunosuppressive component (ISF) on the primary and secondary antibody response, induced by soluble and/or corpuscular antigens, was evaluated in the sera obtained at different intervals before and after immunizations. The duration of the immune suppression induced by ISF treatment within the primary and secondary immunizations was also determined.

METHOD OF STUDY: The ability of the seminal immunosuppressive component to suppress the primary and secondary antibody response was evaluated by enzyme‐linked immunoadsorbent assay (ELISA) in the sera of mice treated in vivo with the immunosuppressor before and after immunization with antigens. Likewise, the duration of the immune suppression induced by the seminal immunosuppressor administered before the primary and secondary immunizations was tested by ELISA with antisera to keyhole limpets hemocyanin.

RESULTS: Intravenous and rectal deposition of ISF led to a suppression of the primary and secondary antibody response to soluble and corpuscular antigens. The most effective suppression of the immune response was achieved in mice treated with immunosuppressor 3 days before the immunization with antigens. Also the secondary antibody response to the challenging antigen was significantly suppressed by ISF. The production of immunoglobulin G (IgG), IgM, and IgA to keyhole limpets hemocyanin was depressed for a relatively long period in mice treated with the immunosuppressor. The results indicated that the preexposure is needed for maximal immunosuppression of the primary antibody production. The treatment with ISF led to a prolonged immunosuppression but not to permanent tolerance to the challenging antigen.

CONCLUSIONS: The in vivo deposition of semen may compromise some aspects of the immune system and may be an important factor in the development of viral and bacterial infections. The suppression of humoral immune response suggests potential uses of seminal immunosuppressor for the animal model study in the therapy of antibody‐mediated diseases.
Expression of TJ6 During Pregnancy
Authors: Kang J.A. , McBey B.A. , Angkachatchai V. , Croy B.A. , Beaman K.D.
Published: September 2011

PROBLEM: TJ6 will be one of the molecules involved in fetal‐specific immune suppression during pregnancy. In the mouse and human decidua, the regulation of uterine natural killer (uNK) cells is important during pregnancy.

METHOD OF STUDY: To further understand the possible functions of TJ6 during pregnancy, syngeneic, allogeneic, and mutant mice were examined for TJ6 expression. RESULTS: Immunoblotting showed that TJ6 protein was expressed on most of the placenta‐associated mononuclear cells, and the size was 70–72 kDa at all stages of pregnancy. The expression of TJ6 mRNA was studied by a ribonuclease protection assay in syngeneic and allogeneic matings, and in immune‐deficient mice of genotypes scid/scid and scid/scid. bg/bg.

CONCLUSIONS: Genetic disparity, lack of T and B lymphocytes, and loss of NK lytic function had no significant effect on the expression of TJ6 mRNA.
Suppressive Effect on Lymphoproliferation In Vitro by Soluble Annexin II Released from Isolated Placental Membranes
Authors: Aarli ÅNen , Kristoffersen Einar K. , Jensen Tone Skeie , Ulvestad Elling , Matre Roald
Published: September 2011

PROBLEM: Syncytiotrophoblast microvillous plasma membranes (StMPM) are potent suppressors of lymphoproliferation in vitro. We have previously shown that soluble annexin II (AII) is present at higher levels in retroplacental serum (RPS) than in peripheral serum, and that soluble AII has an immunosuppressive effect. The aims of this study were to determine whether AII can be released from StMPM and whether soluble AII from StMPM exerts any immunosuppressive effect.

METHOD OF STUDY: Isolated StMPM were incubated in growth medium for 18 hr and supernatants were prepared by ultracentrifugation. Soluble AII was detected by immunoblotting. StMPM, StMPM supernatant, and affinity‐purified AII were analysed in a lymphoproliferation assay for immunomodulating activity.

RESULTS: AII heavy chain and its p11 light chain were detected both in StMPM supernatant and in RPS after removal of StMPM particles by ultracentrifugation. StMPM, StMPM supernatant, and purified AII suppressed lymphoproliferation in a dose‐dependent manner. Absorption of AII from StMPM supernatant reduced the suppressive activity. The suppressive effect of StMPM supernatant and purified AII was completely reversed by heating at 100°C for 30 min or by adding recombinant interleukin‐2 at 100 units/ml. Although StMPM and affinity‐purified AII suppressed the proliferation of lymphocytes from all donors tested, StMPM supernatant suppressed the proliferation of lymphocytes from 12 of 23 donors. Six of eight female non‐suppressed donors were multiparae, whereas five of five female suppressed donors were nulliparae.

CONCLUSIONS: Annexin II is released by isolated placental membranes in vitro and is present in RPS, indicating in vivo release of AII at the fetomaternal interface, probably as AII heterotetramer. AII has immunosuppressive activity and may be important in fetal allograft survival.
Induction of Autoimmune Prostatitis Using Liposomes Is Associated to Peritoneal Cells Activation
Authors: Iribarren Pablo , Correa Silvia G. , Riera Clelia M.
Published: September 2011

PROBLEM: Study and characterization of rat peritoneal cells (PC) involved in the induction of autoimmune prostatitis after the intraperitoneal administration of native extract of accessory glands (RAG) associated with liposomes (RAGL).

METHOD OF STUDY: Induction of the autoimmune response in normal recipients by transferring PC or adherent‐PC loaded with RAGL (RAGL‐PC), but not with PC loaded with empty liposomes (L‐PC). Characterization of the morphology, the ultrastructure, and the phenotype of L‐PC or RAGL‐PC. Study of the respiratory burst by the nitroblue tetrazolium (NBT) reduction assay after stimulation with phorbol myristate acetate (PMA) in both L‐PC and RAGL‐PC.

RESULTS: Liposomes attached to the cell surface of the Mφ were observed by electron microscopy. FACS analyses showed a similar staining pattern with high expression of Ia molecules on L‐PC and RAGL‐PC compared with controls. PMA‐stimulated L‐PC or RAGL‐PC markedly reduced the NBT compared with controls.

CONCLUSION: Our results suggest that the effective uptake of liposomes and the initial activation of PC together with a prolonged stimulatory effect help to disrupt the tolerance state. The present experimental model is an interesting approach to further characterize events associated with antigenic presentation when an autoimmune response is triggered.
Post‐thymectomy Murine Experimental Autoimmune Oophoritis Is Associated With Reduced Natural Killer Cell Activity
Authors: Maity Rita , Nair Sharmila R. , Nelson Lawrence M. , Caspi Rachel R.
Published: September 2011

PROBLEM: Natural killer (NK) cells can influence the immune response by secreting potent lymphokines. It has been suggested that NK cells have a suppressive action on B cells, and that impaired NK cell activity may play a role in some types of autoimmunity. NK cell abnormalities have been reported in women with premature ovarian failure. We therefore examined NK cell activity during the development of murine experimental autoimmune oophoritis, which serves as a model for autoimmune ovarian failure in women.

METHOD OF STUDY: Neonatally thymectomized and sham‐operated C57B1/6 × A/J (B6A) mice were prepared and sacrificed at 4, 6, 8, and 10 weeks after surgery. Splenic NK cell activity was determined in groups of five or more mice by measuring the percent specific lysis of target YAC‐1 lymphoma cells using a standard 4‐hr chromium release cytotoxicity assay. The number of splenic NK cells in neonatally thymectomized and sham‐operated animals was also compared using flow cytometry. In a subsequent experiment, interleukin 12 (IL‐12; NK cell‐stimulating factor) was administered to neonatal mice before neonatal thymectomy.

RESULTS: Neonatally thymectomized mice with associated autoimmune oophoritis had a 75% reduction in the number of splenic NK cells, and 50% or greater reduction in splenic NK cell activity at 4, 6, and 8 weeks after surgery. IL‐12 treatment before neonatal thymectomy maintained NK cell activity and was shown to ameliorate the associated autoimmune oophoritis.

CONCLUSION: Murine post‐thymectomy autoimmune oophoritis is associated with reduced NK cell number and impaired NK cell activity, and in these respects the model is similar to premature ovarian failure in women. Research to define the relationship between NK cell abnormalities and the mechanism of ovarian failure in this model might lend insight into the pathogenesis of premature ovarian failure in women.
A Tissue type Transglutaminase in Human Seminal Plasma
Authors: Whyard Terry C. , Ablin Richard J.
Published: September 2011

PROBLEM: Initial studies from this laboratory of human seminal plasma (SePI) permitted the presumptive identification of the participation of transglutaminase (TGase) and prostaglandins as principal molecules contributing to the immunoregulatory activity of SePI. As a step toward confirmation and extension of these studies, SePI TGase has been partially purified, characterized, and localized.

METHOD OF STUDY: An enzyme‐enriched and partially purified preparation of SePI TGase obtained by molecular sieve and ion‐exchange chromatography and the polyclonal antibody to this preparation were characterized by enzymatic analysis and evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE); immunoprecipitation and immunofluorescence (IF).

RESULTS: A Ca2+ ‐dependent, thrombin‐independent, TGase enzyme from SePI was identified. A pH of 7.5 and a concentration of 2 mM calcium chloride were determined to be optimal for ascertaining the SePI TGase activity in a filter paper assay. Immunoprecipitation using polyclonal antibody prepared with a semipurified TGase preparation, concurrently comparing increasing serum dilution and enzyme activity, revealed a predominant protein band on SDS‐PAGE of 83 kDa. IF studies using the polyclonal antibody on human prostate tissue showed reactivity with a variety of epithelial and stromal components. A significant (P < 0.05) correlation between the biochemical, i.e., enzyme activity of the SePI TGase active preparation, and immunologic activity, i.e., indirect IF staining titre of antibody to acinar epithelial cells and luminal contents, was observed.

CONCLUSIONS: The results confirm and extend initial studies of the presumptive identification of a tissue type TGase associated with the immunoregulatory activity of human SePl and permit the characterization and immunohistologic localization of this macromolecule to the prostate. These observations provide a basis for further investigation of the immunoregulatory role of SePl and prostate TGase in the biology of reproduction and associated pathoses.
Cytokine Production by Lymphocytes in Pregnancy
Authors: Szereday L. , Varga p. , Szekeres‐Bartho J.
Published: September 2011

PROBLEM: In the presence of progesterone lymphocytes of pregnant women release a 34‐ kDa protein named the progesterone‐induced blocking factor (PIBF). PIBF mediates the immunomodulatory and anti‐abortive effects of progesterone and its presence is related to the outcome of pregnancy. PIBF induces production of Th2 type cytokines by activated lymphocytes. The in vivo relationship between PIBF‐ and cytokine production of pregnancy lymphocytes and the outcome of pregnancy was investigated.

METHOD OF STUDY: Interleukin (IL)‐12 and IL‐10 production and PIBF expression in peripheral lymphocytes of 111 healthy pregnant women and 120 women at risk for premature pregnancy termination were detected by immunocytochemistry. RESULTS: We found increased IL‐12 and low PIBF and IL‐10 expression on lymphocytes of “risk” patients, and a high rate of IL‐10 and PIBF positivity on lymphocytes from healthy pregnant women. The cytokine production pattern of the lymphocytes was related to the presence or absence of previous abortions as well as to the outcome of pregnancy. CONCLUSION: These data suggest the involvement of an altered cytokine production pattern in the immunologic effects of progesterone.
Regulation of Decidual Cell and Chorion Cell Production of Interleukin‐10 by Purified Bacterial Products
Authors: Dudley Donald J. , Edwin Samuel S. , Dangerfield Albina , Jackson Kirsten , Trautman Michael S.
Published: September 2011

PROBLEM: To determine whether cultured human decidual cells and chorion cells produce interleukin‐10 (IL‐10) after incubation with purified bacterial products.

METHOD OF STUDY: Decidual cell cultures and chorion cell cultures were established by standard techniques. With confluence, monolayers of each culture were incubated with purified bacterial products, including various concentrations of lipopolysaccharide (LPS), lipid A, and lipoteichoic acid (LTA) for 16 hr in quadruplicate. Culture supernatants were collected and assayed for immunodetectable IL‐10 by enzyme‐linked immunoadsorbent assay (ELISA).

RESULTS: Both decidual cell cultures and chorion cell cultures produced significant quantities of IL‐10 after stimulation with LPS, lipid A, and LTA. Cultures of decidual cells produced more IL‐10 than did chorion cell cultures.

CONCLUSIONS: Our data indicate that both maternal decidual cells and fetally derived chorion cells can produce IL‐10 after incubation with bacterial virulence factors. This finding contrasts with our previous findings in which chorion cells did not produce IL‐10 after stimulation with IL‐1β, suggesting that chorion cell production after incubation with bacterial products is independent of IL‐1β. We speculate that the contribution of anti‐inflammatory IL‐10 production by human gestational tissues to the inflammatory process in these tissues may be overcome or abrogated by the pro‐inflammatory process.
The Effect of Interleukin‐1β and Interleukin‐4 on the Expression of Prostaglandin Receptors EP1 and EP3 in Amnion WISH Cells
Authors: Spaziani Eric P. , Tsibris John C. M. , Hunt Lois T. , Benoit Raymond R. , O'Brien William F.
Published: September 2011

PROBLEM: Although prostaglandin E2 (PGE2) is believed to modulate biochemical and immunological events leading to parturition, the role of prostaglandin E receptors during labor has not been investigated.

METHOD OF STUDY: Amnion WISH cells were incubated in media containing increasing concentrations of either interleukin‐1β (IL‐1β) or IL‐4. Increased EP1 and EP3 protein expression was determined by Western blot analysis with peptide‐specific antibodies. Concomitant measurements of culture media PGE2 were made by an enzyme immunoassay.

RESULTS: Incubation of WISH cells with IL‐1β or IL‐4 caused a two‐to three‐fold increase in EP1 protein levels. IL‐1β and IL‐4 also caused six‐and two‐fold increases, respectively, in culture fluid PGE2 concentrations. IL‐1β or IL‐4 had no effect on EP3 protein levels.

CONCLUSIONS: Based on these results, it is proposed that IL‐1β and IL‐4 may be involved in the initiation and promotion of labor by inducing EP1 levels and PGE2 production in amnion.
Vascular Endothelial Growth Factor Is Increased in Patients With Preeclampsia
Authors: Kupferminc Michael J. , Daniel Yair , Englender Talma , Baram Amiram , Many Ariel , Jaffa Ariel J. , Gull Ilan , Lessing Joseph B.
Published: September 2011

PROBLEM: This study was conducted to determine whether altered levels of vascular endothelial growth factor (VEGF) may play a role in the pathogenesis of preeclampsia.

METHOD OF STUDY: Maternal plasma samples were collected from 19 patients with preeclampsia (group A) either before the onset of labor, or before induction of labor or medical intervention. Plasma samples were also obtained from 19 normotensive patients with uncomplicated pregnancies (group B), who were matched with the patients with preeclampsia for gestational age and parity. Samples were frozen at −70°C until assayed for VEGF by a specific enzyme‐linked immunoassay.

RESULTS: The mean maternal age was similar in groups A and B. For both groups the VEGF was detectable in all plasma samples. However, the plasma concentrations of VEGF were significantly increased in the group A patients, compared with those in group B (median, 47ng/ml; range, 10.6–72 ng/ml versus median, 13.6 ng/ml; range, 0.66‐20 ng/ml; P < 0.001). In group A, a positive correlation was noted between VEGF concentrations and the systolic and diastolic blood pressure (r = 0.56; P = 0.01 and r = 0.48; P = 0.037, respectively).

CONCLUSIONS: Maternal plasma VEGF levels were elevated in the patients with preeclampsia and correlated with the severity of hypertension, suggesting a role for VEGF in the pathogenesis of preeclampsia.