bluuebunny/crossref_metadata_2025 huggingface dataset:
Recent Studies on the Structure of the grc Region in the Rat
Authors: Gill Thomas J. , Yuan Xiu‐Juan , Salgar Shashikumar K. , Kunz Heinz W.
Published: September 2011

PROBLEM: To explore the nature of the genes in the grc.

METHOD: Chromosome walking of the grc and sequencing new genes.

RESULTS: The RT1.O, N genes have been aligned, and two new types of genes, RT1.S1,2 and Rpr1,2,3, have been discovered and mapped. An extensive physical map of the grc region is presented.

CONCLUSION: The disease associations in both the rat and the human appear to cluster in two regions that map in approximately the same place in both species, given the translocation that occurred in the major histocompatibility complex (MHC) of the rat relative to the MHC of the human.
GRO‐alpha in Human Serum: Differences Related to Age and Sex
Authors: Kanda Yuki , Koike Dr. Koji , Sakamoto Yoshiki , Osako Yasuko , Masuhara Kanji , Watanabe Kazuyoshi , Tsurufuji Susumu , Hirota Kenji , Miyake Akira
Published: September 2011

PROBLEM: Human GRO‐alpha (GRO‐α) is a new member of the chemokine family that is supposed to play an important role in inflammatory and immune reactions. We established a sandwich enzyme‐linked immunoassay (ELISA) system with polyclonal antibodies against human GRO‐α and investigated the serum level of healthy donors to establish normal ranges for this chemokine in adults.

METHODS: GRO‐α concentrations were measured cross‐sectionally in the sera of 240 healthy adults. The variability of serum GRO‐α levels was also measured in normal volunteers, samples from whom were obtained by sequential venipunctures or by a small plastic cannula with a heparin‐saline lock, to determine short‐term variability.

RESULTS: Whereas there was no difference between the concentration of human GRO‐α from men (logarithmic mean, 77.6 pg/ml, n = 120) and that from women with normal menstrual cycles (log mean, 71.6 pg/ml, n = 73), the concentration from postmenopausal women (log mean 45.0 pg/ml, n = 31) was lower than that from women with normal menstrual cycles (log mean 71.6 pg/ml, n = 73). However, we could not detect any significant difference between healthy donors' serum levels and those of donors with acute inflammation. Fewer variations were recognized in the case of the sequential venipunctures method than in that of the heparin‐saline lock method.

CONCLUSION: We found that the GRO‐α concentration of postmenopausal women was significantly lower than that of women with normal menstrual cycles. These results suggest the GRO‐α serum levels of normal healthy women may have some correlation with sex hormones.
Pathogenetic Implication of Interleukin‐2 Expressed in Preeclamptic Decidual Tissues: A Possible Mechanism of Deranged Vasculature of the Placenta Associated with Preeclampsia
Authors: Hamai Yoko , Fujii Tomoyuki , Yamashita Takahiro , Kozuma Shiro , Okai Takashi , Taketani Yuji
Published: September 2011

PROBLEM: The purpose of this study is to clarify whether IL‐2 expressed in the decidua in preeclampsia affects the angiogenesis of the placenta.

METHOD OF STUDY: We investigated the angiogenic substances released from human trophoblasts obtained from early pregnancy that had been pretreated with either IL‐2, non‐activated lymphocytes from peripheral blood mononuclear cells (PBMCs), decidual lymphocytes, or these lymphocytes activated by lymphokine (LAK cells). Angiogenic activity was determined by evaluating the ability of growth‐promotion of cultured human microvascular endothelial cells (HMvECs) using MTT assay.

RESULTS: Trophoblasts pretreated with IL‐2 or non‐activated lymphocytes, irrespective of their origin, released angiogenic factor similar to those without pretreatment. However, trophoblasts pretreated with LAK cells released less angiogenic factor compared with those without pretreatment.

CONCLUSIONS: Interleukin‐2 (IL‐2) expressed in preeclamptic decidua might reduce the angiogenic substances arising from trophoblasts by inducing LAK cells from decidual lymphocytes, which might be relevant to deranged vasculature of the placenta, a characteristic histology in preeclampsia.
TGF‐β2 and PGE2 in Rabbit Blastocoelic Fluid Can Modulate GM‐CSF Production by Human Lymphocytes
Authors: Fortin Marylène , Ouellette Marie‐Josée , Lambert Raymond D.
Published: September 2011

PROBLEM: During normal pregnancy, major changes occur in the production of Th2/Th1 cytokines at the feto‐maternal interface. Th2 cytokines such as interleukin‐4 (IL‐4) or interleukin‐10 (IL‐10) are predominantly produced locally in the uterine and placental tissues, whereas the production of Th1 cytokines such as tumor necrosis factor alpha (TNF‐α) and interferon gamma (IFN‐γ) are decreased. Because these modulations might be induced by the embryo, the current study was carried out to test the effect of rabbit blastocoelic fluid on the production of Th2/Th1 cytokines by lymphocytes, and to investiate the possible implication of transforming growth factor β2 (TGF‐β2) prostaglandin E2 (PGE2) as modulators of the production of these cytokines.

METHOD OF STUDY: Human peripheral blood lymphocytes (PBL) were cultured along with ConcanavalinA (Con A), and rabbit blastocoelic fluid was collected on day 12 of gestation (BF d‐12). Concentrations of cytokines in culture media were determined by enzyme‐linked immunoadsorbent assay (ELISA).

RESULTS: Addition of BF d‐12 in the culture medium induced a strong inhibition of IL‐2, TNF‐α, IL‐10, and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) production. However, an initial pretreatment of the lymphocytes with BF d‐12, followed by a Con A stimulation, led to a marked increase in GM‐CSF production, whereas IL‐2, TNF‐α, and IL‐10 secretions were inhibited. It was also demonstrated, for the first time, that a pretreatment of the lymphocytes with TGF‐β2 and PGE2 increased GM‐CSF production to the same level reached after the addition of BF d‐12. Furthermore, removal of TGF‐β2 and PGE2 from BF d‐12 by affinity chromatography reduced the effect of BF d‐12 on GM‐CSF production.

CONCLUSIONS: Taken together, these findings suggest that the embryo, in modulating harmful and beneficial cytokine production locally, plays an active role in its protection against maternal immune cellular assault. These results also emphasize the importance of growth factors for successfully maintaining pregnancy.
Genetics of Reproduction and Its Evolutionary Significance
Authors: Gill Thomas J.
Published: September 2011

PROBLEM: To provide an evolutionary perspective on reproductive problems.

METHOD OF STUDY: A broad range of evolutionary and reproductive data in the literature were analyzed.

RESULTS: Evolutionary considerations provide some insight into two aspects of human reproduction. First, the inefficiency of the reproductive process (20–35% of fertilized ova yield successful pregnancies) may reflect the stress placed on the reproductive process by the relatively short evolutionary time between the hunter‐gatherer and settled agricultural patterns of human life. Second, the immaturity of the immune response of the human newborn is most likely a consequence of its delivery while still in the embryonic/fetal stage of development. Delivery at this time is necessitated by the rapid development of the human brain in utero.

CONCLUSIONS: The attempts to develop reproductive technologies should take into account the fundamental limits on the human reproductive process. A variety of in utero preventive and therapeutic techniques should be developed to provide the newborn with enhanced resistance to infectious diseases or to correct congenital defects. All new procedures should be thoroughly investigated and proved before being put into clinical practice.
Immunogenetic and Oncogenic Properties of Rat Trophoblast Cell Lines
Authors: Kunz Heinz W. , Yuan Xiu‐Juan , Salgar Shashikumar K. , Dixon‐McCarthy Barbara , Gill Thomas J.
Published: September 2011

PROBLEM: The nature of major histocompatibility complex (MHC) antigen expression on rat placentas, trophoblast cell lines, and tumors derived from trophoblast cells was explored.

METHOD OF STUDY: Cytohistochemical and flow cytometric analysis and molecular techniques.

RESULTS: MHC antigen expression and genomic imprinting on the placenta and on trophoblast cells varies with the time of gestation and with the type of MHC antigen.

CONCLUSIONS: There is no correlation in trophoblast cells between class I expression and cell ploidy, on the one hand, and malignant potential, on the other hand. Genomic imprinting of class I antigens in the rat placenta is a quantitative phenomenon.
Progesterone and Non‐specific Immunologic Mechanisms in Pregnancy
Authors: Szekeres‐Bartho Julia , Par G. , Szereday L. , Achatz I. , Smart C.Y.
Published: September 2011

PROBLEM: Progesterone‐dependent immunomodulation is one of the mechanisms that enables pregnancy to proceed to term. Immunologic effects of progesterone are mediated by a protein named the progesterone‐induced blocking factor (PIBF). Among other effects this protein inhibits natural killer (NK) activity and displays an anti‐abortive effect in mice. Recently we have shown that PIBF induces a Th2 shift in vitro. The present study was aimed at investigating the in vivo effect of PIBF on cytokine production, as well as the relationship between cytokine production, NK activity, and pregnancy loss.

METHOD OF STUDY: Balb‐c mice on day 8.5 of pregnancy were injected intraperitoneally with 0.5 mg of rabbit anti‐PIBF immunoglobulin G (IgG). Another group of mice was simultaneously treated with anti‐NK monoclonal antibodies. Mice treated with the same amount of normal rabbit serum or untreated mice of similar gestational age were used as controls. The animals were sacrificed on day 10.5, and their uteri were inspected. The ratio of living and resorbed embryos was determined. NK activity as well as cytokine expression on the spleen cells were determined by immunocytochemistry and enzyme‐linked immunoadsorbent assay (ELISA).

RESULTS: Mitogen‐activated spleen cells from anti‐PIBF‐treated mice produced significantly (P < 0.001) less IL‐10 than those of pregnant control mice. A significantly higher percentage (P < 0.001) of spleen cells from anti‐PIBF‐treated mice expressed interferon‐γ (IFNγ) as determined by immunocytochemistry, than those of untreated pregnant mice. There was a positive relationship between the percentage of IFNγ‐positive spleen cells and resorption rates, and an inverse relationship between the latter and interleukin‐10 (IL‐10) production. All these effects were corrected by treatment with anti‐NK antibodies. CONCLUSION: Our data suggest that PIBF contributes to the success of gestation via eytokine‐mediated inhibition of NK activity.
Endothelial Cells as a Target for Antiphospholipid Antibodies: Role of Anti‐Beta 2 Glycoprotein I Antibodies
Authors: Del Papa N. , Raschi E. , Catelli L. , Meroni P.L. , Khamashta M.A. , Ichikawa K. , Tincani A. , Balestrieri G.
Published: September 2011

PROBLEM: To investigate the role of the plasma cofactor for antiphospholipid antibodies in antibody binding to endothelial cells.

METHOD OF STUDY: a) Evaluation of endothelial cell binding of polyclonal and monoclonal anti‐β2 glycoprotein I antibodies; and b) study of the effects of antibody binding: adhesion molecule expression, leukocyte adhesion, and interleukin‐1β secretion. RESULTS: Anti‐β2 glycoprotein I antibodies bind endothelial cell monolayers in vitro by reacting with the cofactor adhered to the cell membranes. Antibody binding induces an upregulation of adhesion molecules, favours leukocyte adherence, and increases interleukin‐1β secretion. Interleukin‐1β plays an active role to mediate adhesion molecule expression through an autocrine loop, as shown by the inhibitory effect of interleukin 1 receptor antagonist.

CONCLUSIONS: Antiphospholipid antibodies do react with endothelium through the co‐factor adhered to their cell membranes and induces a pro‐adhesive cell phenotype.
Paternal Leukocytes Selectively Increase Secretion of IL‐4 in Peripheral Blood During Normal Pregnancies: Demonstrated by a Novel One‐Way MLC Measuring Cytokine Secretion
Authors: Ekerfelt Christina , Ernerudh Jan , Matthiesen Leif , Berg Göran
Published: September 2011

PROBLEM: It has been proposed that immune responses in normal pregnancy are Th2‐like, thereby protecting the fetus and placenta from being rejected. Some studies have shown Th2‐deviated systemic responses to different antigens and mitogens. The aim of this study was to demonstrate the specific T cell cytokine responses directed toward paternal histocompatibility leukocyte antigen (HLA), because this is the most prominent target for rejection of the feto‐placental unit.

METHOD OF STUDY: A novel one‐way mixed leukocyte culture (MLC) combined with the detection of cytokine secretion with a sensitive ELISPOT assay was developed. Peripheral blood from 11 pregnant women was investigated with respect to allo‐reactivity toward paternal leukocytes and pooled leukocytes from unrelated blood donors. This was done at three different occasions during pregnancy and 8 weeks after delivery. Nine age‐matched non‐pregnant women served as controls.

RESULTS: In the second and third trimesters of pregnancy significantly larger numbers of IL‐4‐secreting cells (Th2) were induced by paternal leukocytes as compared to unrelated leukocytes.

CONCLUSIONS: The findings indicate a selective immune deviation toward Th2, which may protect the fetus from rejection and thus may be an important homeostatic mechanism in normal pregnancies.
Stability of Serum Interleukin‐10 Levels During the Menstrual Cycle
Authors: Maskill Joanna K. , Laird Susan M. , Blakemore Alexandra I. F. , Okon M. , Li T. C.
Published: September 2011

PROBLEM: Menstrual cycle‐associated variability in the circulating levels of several cytokines can be a confounding factor in measurements of in vivo cytokine levels in clinical studies. Since pregnancy‐associated increases in interleukin‐10 (IL‐10) levels are well documented, we have investigated the variability in serum levels of IL‐10 in healthy women at different stages of the menstrual cycle to ascertain whether this is a problem in comparative studies of circulating IL‐10 levels.

METHOD OF STUDY: We obtained fifty‐four successive serum samples at points in the menstrual cycles of 12 healthy fertile women, precisely timed by measurement of the luteinizing hormone surge, and measured the interleukin‐10 levels.

RESULTS: Levels of IL‐10 in successive serum samples from each woman taken on days LH‐7 (that is seven days prior to LH surge), LH‐4, LH+1, LH+7, and LH+10 showed that IL‐10 does not vary in a systematic way during the menstrual cycle.

CONCLUSION: These results validate the sampling of women in studies of IL‐10 levels in various clinical situations and establish that these levels are not dependent on menstrual cycle dates. They also suggest that menstrual cycle‐related changes in IL‐1 are not mediated by IL‐10. The rise in progesterone in the luteal phase of the menstrual cycle is not mirrored by a rise in the circulating IL‐10 level, which implies either that the pregnancy‐associated rise is not related to progesterone or that it is only observed at the higher progesterone levels in pregnancy.
Peritoneal Cellular Immunity and Endometriosis
Authors: Ho Hong‐Nerng , Wu Ming‐Yih , Yang Yu‐Shih
Published: September 2011

PROBLEM: An immunologic basis has long been considered to be very important in the pathogenesis of endometriosis. Interactions of the peritoneal cells, which comprise macrophages, B cells, T cells, natural killer (NK) cells, and retrograde endometrial cells, are critical, but remain controversial, for exploring the pathogenesis of endometriosis. METHOD OF STUDY: Accumulated data from the literature were reviewed, and our data were analyzed.

RESULTS: The data show that peritoneal macrophages are activated by the recurrent reflux of menstrual shedding. Humoral and local endometrial autoantibodies are detected in patients with endometriosis, but B cells are not quantitatively increased. There is decreased NK cell activity in the peritoneal cavity and peripheral blood, and this decreased activity may be related to the failure to clear out the ectopic endometrial tissue. Peritoneal T cells are predominant by Thl inflammatory cells, and these cells are impaired because of a decrease in activation (especially HLA‐DR+ CD4+ CD3+ population) and in the production of interleukin‐2. Inflammatory cytokines such as interleukin‐1, interleukin‐6, and tumor necrosis factor‐α are elevated in the peritoneal fluid of women with endometriosis. CONCLUSIONS: The peritoneal NK and T lymphocytes are suppressed in women with endometriosis, but whether these immunologic deviations are the cause or the result of endometriosis is still unclear. Further studies are required to determine what role immunologic factors play in the pathophysiology of endometriosis.
Transferrin Receptor (CD71) Expression in Peritoneal Macrophages from Fertile and Infertile Women With and Without Endometriosis
Authors: Martínez‐Román Sergio , Balasch Juan , Creus Montserrat , Fábregues Francisco , Carmona Francisco , Vilella Ramón , Vanrell Juan A.
Published: September 2011

PROBLEM: Hyperactivated macrophages are implicated in the pathophysiology of endometriosis‐associated infertility. This study investigates transferrin receptor expression (CD71) as a marker of hyperactivity in peritoneal macrophages of infertile patients with minimal to mild endometriosis (group 1, n = 25).

METHOD OF STUDY: Expression of the activation antigen CD71 on peritoneal fluid macrophages was determined by a specific monoclonal anti‐CD71 antibody using indirect immunofluorescence technique and was analyzed by flow cytometry. Three different control groups of women were used: women with unexplained infertility (group 2, n = 25), fertile women with endometriosis (group 3, n =10), and fertile women without endometriosis (group 4, n = 25).

RESULTS: The percentage of CD71 positive cells was significantly increased in infertile women with endometriosis as compared with the three control groups. There were no differences among groups 2, 3, and 4 with respect to the percentage of CD71 positive macrophages.

CONCLUSIONS: Our results favor the concept that hyperactivated macrophages play a role in the pathophysiology of endometriosis‐associated subfertility, a feature which is lacking in patients with unexplained infertility.
Soluble Human Leukocyte Antigens, Interleukin‐6, and Interferon‐γ During Pregnancy
Authors: Russwurm G.P. , Mackler A.M. , Fagoaga O.R. , Brown W.S. , Sakala E.P. , Yellon S.M. , Nehlsen‐Cannarella S.L.
Published: September 2011

PROBLEM: Soluble human leukocyte antigens (sHLA), interferon‐γ (IFN‐γ), and interleukin‐6 (IL‐6) were studied during human pregnancy to test the hypothesis that sHLA concentrations are regulated by these specific cytokines.

METHOD OF STUDY: Enzyme‐linked immunoadsorbent assays (ELISA) were used to measure sHLA I and II in maternal circulation, cord blood, and placenta effluents of pregnant and nonpregnant women; maternal serum cytokines were also determined.

RESULTS: sHLA in maternal and cord blood were equivalent to that in the placenta. By the third trimester, sHLA I concentrations in maternal plasma were significantly reduced compared to the first or second trimesters. sHLA II was increased during the second trimester relative to that postpartum. Maternal IL‐6 and IFN‐γ concentrations were not statistically different throughout gestation or postpartum.

CONCLUSIONS: These data do not suggest a role for maternal plasma IL‐6 or IFN‐γ in regulation of systemic sHLA class I during pregnancy, but they do not address whether such events take place in local tissues of the maternal‐fetal unit.
Lethal Outcome of Uterine Infection in Pregnant but Not in Nonpregnant Rats and Increased Death Rate With Inhibition of Nitric Oxide
Authors: Nowicki B. , Fang L. , Singhal J. , Nowicki S. , Yallampalli C.
Published: September 2011

PROBLEM: Limited information is available on potential differences in sensitivity to urogenital infections between pregnant and nonpregnant hosts.

METHOD OF STUDY: In this study, we evaluated Escherichia coli infectious complications in pregnant and nonpregnant rats and the effect of nitric oxide (NO) inhibitor, NG‐nitro‐l‐arginine methyl ester (l‐NAME), on the outcome of an experimental uterine infection.

RESULTS: Of the infected pregnant animals, 31% were found dead in 24–48 hr. The death rate was increased 2‐fold (66%) with l‐NAME treatment. No deaths occurred in nonpregnant animals with or without l‐NAME treatment. The rate of uterine infection in pregnant animals was about 10‐fold higher than in nonpregnant animals.

CONCLUSION: We propose that infectious complications of pregnancy may be related to gestation‐dependent sensitivity to the pathogenic microorganism and the host NO status.
Cytokine Expression by First‐Trimester Human Chorionic Villi
Authors: Bennett W.A. , Lagoo‐Deenadayalan S. , Stopple J.A. , Barber W.H. , Hale E. , Brackin M. N. , Cowan B. D.
Published: September 2011

PROBLEM: Communication at the human maternal‐fetal interface occurs by an intricate cytokine network. This study examines cytokine expression by normal first‐trimester human chorionic villi.

METHOD OF STUDY: Tissues were obtained at elective pregnancy terminations (7–9 weeks). Total RNA was isolated from chorionic villi by guanidinium isothiocynate‐acid phenol extraction. A reverse transcriptase‐polymerase chain reaction technique was used to examine cytokine expression. β‐Actin was used as the housekeeping gene, and mitogen‐stimulated lymphocytes served as positive controls.

RESULTS: β‐Actin was uniformly expressed by all chorionic villous samples. Interferon (IFN)‐α and ‐β also were highly expressed. Moderate expression was noted for interleukin (IL)‐10, IL‐6, tumor necrosis factor (TNF)‐α, and IL‐1β. In contrast, transforming growth factor‐β1, IFN‐γ, IL‐2, and IL‐1α were either weakly expressed or absent in first‐trimester villi. CONCLUSIONS: Cytokines may contribute to pregnancy immunotolerance (IFN‐α, IFN‐β, and IL‐10), viral resistance (IFNs), hormone secretion (IL‐1 and IL‐6), and cellular remodeling (IFN‐γ and TNF‐α) within the chorionic villous.
Transport of Proteins Across the Human Placenta
Authors: Malek Antoine , Sager Ruth , Schneider Henning
Published: September 2011

PROBLEM: The transport of various proteins across the human placenta was investigated by comparing maternal and fetal concentrations of tetanus antigen (TT‐AG), anti‐tetanus (TT)‐immunoglobulin G (IgG) (following maternal vaccination), IgA, human chorionic gonadotropin (hCG), human placental lactogen (hPL), and alpha‐fetoprotein (AFP) at term.

METHOD OF STUDY: The concentrations of the six proteins were determined using enzyme‐linked immunosorbent assay in serum of maternal venous and umbilical (fetal) vein samples obtained at delivery from uncomplicated term pregnancies (n = 16).

RESULTS: The ratios (mean ± standard deviation) of fetal (umbilical) to maternal level were 1.41 ± 0.33 (anti‐TT‐IgG), 0.91 ± 0.37 (TT‐AG), 0.002 ± 0.001 (IgA), 0.003 ± 0.001 (hCG), and 0.008 ± 0.004 (hPL), while the maternal:fetal concentration ratio of AFP was 0.002 ± 0.002. IgA, hCG, hPL, and AFP showed a close correlation between maternal and fetal levels varying between r2 = 0.47 to 0.73 (P < 0.004–0.0001). Because AFP is produced by the fetus while IgA originates in the mother, the appearance of small amounts of these two proteins in the maternal or fetal compartment, respectively, suggests a slow rate of diffusion following a high concentration gradient. The detection of hCG and hPL in fetal serum is also interpreted as diffusion from the maternal into the fetal blood. Anti‐TT‐IgG has a significantly higher concentration in the fetal as compared with the maternal serum, which is in line with the well‐documented active transfer of IgG. Fetal TT‐antigen levels were similar to maternal concentrations, showing a close correlation (r2 = 0.74, P < 0.0001) between the two proteins.

CONCLUSIONS: The correlation between maternal and fetal concentrations of various proteins like IgA (150,000 Da), hCG (42,000 Da), and hPL (21,000 Da) suggests passive diffusion of these macromolecules across the placenta from the maternal to the fetal side, albeit at a slow rate. A similar process is postulated for AFP (70,000 Da) diffusing in the opposite direction from the fetus to the mother. There was no significant difference between the transplacental fetomaternal gradient of IgA and hCG and the maternal‐fetal gradient of AFP. In view of the substantially larger volume of circulating maternal as compared with fetal blood, a significantly higher rate of crossing of AFP as compared with the other proteins must be assumed. It is uncertain whether a difference in the rate of transplacental transfer in the two directions or an additional source of AFP production in the maternal compartment explains the high maternal level. Anti‐TT‐IgG concentration is significantly higher in fetal than in maternal serum suggesting active transfer from the mother to the fetus. Furthermore, there is considerable transfer of TT‐AG and a close correlation of fetal:maternal ratios of anti‐TT‐IgG (150,000 Da) and TT‐AG (150,000 Da) could be an indication for a specific transfer of the antigen antibody complex.
Endothelin‐1: Expression and Role in Human Corpus Luteum
Authors: Apa Rosanna , Miceli Fiorella , Feo Daniela De , Pierro Emilia , Ayala Gianfranco , Mancuso Salvatore , Napolitano Maddalena , Lanzone Antonio
Published: September 2011

PROBLEM: Several recent data suggest an involvement of endothelin (ET)‐1, a powerful vasoconstrictor peptide, in reproductive function. This study was designed to investigate the presence and role of ET‐1 in human corpus luteum.

METHOD OF STUDY: Purified luteal cells were incubated for different times with ET‐1 or ET‐3 alone or associated with human chorionic gonadotropin. In another set of experiments cells were treated with ET‐1 and BQ485, an ET‐A receptor antagonist, or with phorbol 12‐myristate‐13 acetate (PMA), an activator of protein kinase C.

RESULTS: ET‐1 reduced both basal and human chorionic gonadotropin‐induced progesterone production at all examined times, similarly PMA inhibited basal progesterone synthesis. BQ485 prevented the inhibitory effect of ET‐1, while no effect was observed with ET‐3. Finally, ET‐1 mRNA was detected in the luteal cells.

CONCLUSION: ET‐1 is expressed by human luteal cells and reduces basal and human chorionic gonadotropin‐induced progesterone synthesis through the ET‐A receptors and the protein kinase C pathway. Conversely, ET‐3 does not affect luteal steroidogenesis.
Incidence of Rat‐Soluble Placental Factors on IgE and IgG2a Synthesis
Authors: Gentile Teresa , Dokmetjian Jose , Garrido Delia , Margni R.A.
Published: September 2011

PROBLEM: The in vivo effect of soluble factors present in placental culture supernatants (PCSs) on the synthesis of rat immunoglobulin E (IgE) and IgG2a isotypes was investigated.

METHOD OF STUDY: Batches of Wistar SPF rats immunized with a 10‐μg dose of ovalbumin and Al(OH)3 were used: group I, consisted of virgin rats; group II, virgin females injected simultaneously with PCSs; and group III, pregnant females. As controls, nonimmunized batches were included. Serum samples were collected at days 0 (basal) and 10 after antigen challenge, determining levels of total and specific antiovalbumin of both IgE and IgG2a by enzyme‐linked immunoadsorbent assay (ELISA).

RESULTS: In vivo and at least at the doses administered, PCSs exert an inhibitory effect on the synthesis of specific and total anti‐ovalbumin IgE during the course of immune response to such challenge. However, PCSs did not modify serum values of total and specific IgG2a.

CONCLUSIONS: These results suggest that PCSs exert selective influence on the synthesis of diverse immunoglobulin isotypes during immune response, through the balance of cytokines synthesized by placental cells.
Human Lymphocytes Obtained from Decidual Tissue Express Killer Activatory Receptors as Well as Killer Inhibitory Receptors: Analysis Using a Single Strand Conformation Polymorphism Method
Authors: Miki Akinori , Ishikawa Yoshihide , Yabe Toshio , Furukawa Hiroshi , Fujii Tomoyuki , Tokunaga Katsushi , Akaza Tatsuya , Tadokoro Kenji , Taketani Yuji , Juji Takeo
Published: September 2011

PROBLEM: The objective was to clarify whether lymphocytes in decidual tissue express only killer inhibitory receptors (KIRs) or both KIRs and killer activatory receptors (KARs).

METHOD OF STUDY: A polymerase chain reaction (PCR)‐single strand conformation polymorphism (SSCP) analysis method, using cDNA as a template, was applied to detect as many natural killer receptors (NKRs) expressed on natural killer (NK) cells as possible and to distinguish between KIRs and KARs. Using this method, we analyzed NKRs on lymphocytes in decidual tissue and maternal peripheral blood mononuclear cells (PBMCs) (n = 10) from the same individuals.

RESULTS: More than 20 SSCP bands were detected for NKRs on both the lymphocytes in decidual tissue and the maternal PBMCs. The SSCP band patterns were different for each individual. KARs mRNA was detected in lymphocytes in decidual tissue according to the SSCP analysis results.

CONCLUSION: Lymphocytes in decidual tissue express not only KIRs but also KARs; this finding suggests their roles in placentation and the maintenance of pregnancy.
Early Recognition of Pregnancy by the Maternal Immune System
Authors: Kelemen K. , Szekeres‐Bartho Julia , Paldi A. , Tinneberg H. , Torok A.
Published: September 2011

PROBLEM: Immunologic recognition of pregnancy is important for normal gestation. Successful pregnancy is characterized by a Th2 dominance, whereas there is a Th1 dominance in failed pregnancies. We assume that a signal given by the fertilized ovum induces a Th2 shift, necessary for a normal outcome. In vitro fertilization provides a tool for testing this possibility.

METHOD OF STUDY: Phytohemagglutinin‐stimulated lymphocytes were incubated for 48 hr in the presence of culture media from in vitro fertilized eggs, as well as in follicular fluid (FF) and control supernatants. Total RNA was isolated from the lymphocytes by the guanidine‐isothiocyanate method and interleukin (IL)‐10 mRNA expression was detected by reverse transcription‐polymerase chain reaction.

RESULTS: Ten percent of the activated lymphocytes incubated with FF expressed IL‐10 mRNA, whereas 88% of the lymphocytes activated with supernatants of sperm + oocytes gave a positive signal. Significantly (P < 0.05) fewer (50%) lymphocytes stimulated in the presence of control supernatants also expressed mRNA for IL‐10.

In supernatants of activated lymphocytes incubated with the culture medium of spermia + oocytes, the concentration of IL‐10 was significantly higher than in the lymphocytes incubated with FF.

CONCLUSION: These data suggest that the presence of the fertilized ovum induces a Th2 shift, which enables pregnancy to proceed to term.
Plasma Interleukin‐12 Is Elevated in Patients with Preeclampsia
Authors: Daniel Yair , Kupferminc Michael J. , Baram Amiram , Jaffa Ariel J. , Fait Gideon , Wolman Igal , Lessing Joseph B.
Published: September 2011

PROBLEM: Abnormal immune activation has been suggested as a contributor to the development of preeclampsia. We hypothesized that intact interleukin (IL)‐12 directly, or through its main mediator, interferon (IFN)‐γ, contributes to the altered immune response observed in preeclampsia.

METHOD OF STUDY: Plasma samples were collected from 20 patients with preeclampsia and 20 normotensive patients with uncomplicated pregnancies who were matched with the preeclamptic patients by age, gestational age, and parity. Samples were collected before the onset of labor, induction, or medical intervention. The samples were assayed for IL‐12 and IFN‐γ by specific enzyme‐linked immunoassays.

RESULTS: IL‐12 was detected in 35% of the preeclamptic patients and in 5% of the patients with normal pregnancies (P < 0.01). The detection rate and mean concentration of IFN‐γ were comparable in both groups.

CONCLUSION: Intact plasma IL‐12 is detected more frequently in preeclamptic patients, suggesting the involvement of this cytokine in the enhanced immune response observed in preeclampsia.
A Rational Basis for the Use of Combined Heparin/Aspirin and IVIG Immunotherapy in the Treatment of Recurrent IVF Failure Associated with Antiphospholipid Antibodies
Authors: Sher Geoffrey , Zouves Christo , Maassarani Ghanima , Feinman Michael , Matzner William , Chong Penny , Ching Wendell
Published: September 2011

PROBLEMS: 1) Does the administration of heparin and aspirin (H/A) in combination with intravenous immunoglobulin G (IVIG) improve in vitro fertilization (IVF) implantation and birth rates in patients with recurrent IVF failures? 2) Is the effect of such treatment related to the antiphospholipid antibody (APA) status of the patients concerned?

METHOD OF STUDY: Subjects consisted of 89 women younger than 36 years of age whose infertility was a result of causes other than male infertility and who had experienced four or more failed IVF/embryo transfer procedures. Fifty‐two women were APA+ (group A), and 37 were APA‐ (group B). All patients, regardless of their APA status, received H/A (5000 U sq bid), aspirin (81 mg po qd) from the inception of menotropin therapy along with IVIG (20 g) through a single infusion 3 to 10 days before egg retrieval.

RESULTS: Twenty‐two (42%) of group A and 7 (19%) of group B patients achieved live births (P = 0.020).

CONCLUSIONS: IVF outcome is significantly improved when H/A and IVIG are administered to APA+ women with repeat IVF failures. APA‐ women do not seem to benefit from such treatment.
Presence of Leukemia Inhibitory Factor and Interleukin‐12 in Human Follicular Fluid During Follicular Growth
Authors: Coskun Serdar , Uzumcu Mehmet , Jaroudi Kamal , Hollanders Johannes M.G. , Parhar Ranjit S. , Al‐Sedairy Sultan T.
Published: September 2011

PROBLEM: Cytokines have been shown to be present in human follicular fluid and have regulatory functions on follicular maturation. The presence of leukemia inhibitory factor (LIF) and interleukin (IL)‐12 in human follicular fluid obtained at different stages of maturation was investigated.

METHOD OF STUDY: Follicular fluids and granulosa cells were obtained from preovulatory and immature follicles. Follicular fluids from both groups were assayed for IL‐12 and LIF by enzyme‐linked immunosorbent assay. Granulosa cells from preovulatory and immature follicles were treated with human chorionic gonadotropin (hCG) in vitro and subsequent LIF and IL‐12 production were measured.

RESULTS: The average concentration of LIF was significantly higher in preovulatory follicles (7.6 ± 1.3 pg/ml, n = 24) than in immature follicles (2.0 ± 1.3 pg/ml, n = 6). The concentration of IL‐12 was significantly higher in follicular fluid obtained from immature follicles (10.9 ± 5.0 pg/ml) than in preovulatory follicles (1.3 ± 0.4 pg/ml). hCG only stimulated LIF production from mature granulosa cells; it had no effect on IL‐12 production.

CONCLUSIONS: IL‐12 and LIF are present in follicular fluid and their levels are regulated differently during follicular maturation. hCG stimulates LIF production from granulosa cells in vitro.
Interferon‐α is a Potent Inhibitor of Basic Fibroblast Growth Factor‐Stimulated Cell Proliferation in Human Uterine Cells
Authors: LEE BYUNG‐SEOK , STEWART ELIZABETH A. , SAHAKIAN MARINE , NOWAK ROMANA A.
Published: September 2011

PROBLEM: Abnormal uterine bleeding is a significant health problem for many women and is the number‐one reason for performing hysterectomy in the United States. Leiomyomas (uterine fibroids) are benign neoplams that are a frequent cause of abnormal uterine bleeding. The goal of this study was to assess the effects of the anti‐angiogenic cytokine, interferon (INF)‐α, on the proliferation of both leiomyoma and normal uterine cells.

METHOD OF STUDY: Primary cultures of leiomyoma, myometrial, and endometrial stromal cells were established for in vitro study. The effects of INF‐α (10, 100, and 1000 U/ml) were tested on serum‐stimulated and basic fibroblast growth factor‐stimulated cell proliferation using the [3H]thymidine incorporation assay.

RESULTS: INF‐α was a potent inhibitor of cell proliferation for all three cell types, with endometrial stromal cells showing the greatest sensitivity. The antiproliferative effect did not appear to result from toxic effects on the cells.

CONCLUSION: INFs may prove to be useful therapeutic agents for the treatment of leiomyoma‐related abnormal uterine bleeding.
H‐Y Antigen Expression Patterns in Human X‐ and Y‐Chromosome‐Bearing Spermatozoa
Authors: SILLS E. SCOTT , KIRMAN IRENA , COLOMBERO LILIANA T. , HARIPRASHAD JUNE , ROSENWAKS ZEV , PALERMO GIANPIERO D.
Published: September 2011

PROBLEM: Restricted expression of H‐Y antigen on Y‐chromosome‐bearing sperm has been reported in some species, although such preferential expression for H‐Y antigen in human sperm has yet to be described. In this study, an immunomagnetic approach was used to characterize antigen expression patterns as a function of sex‐chromosome content.

METHOD OF STUDY: Human sperm was treated with monoclonal immunoglobulin (Ig) M antibodies directed against H‐Y antigen. This preparation then was incubated with sheep antimouse IgM antibody affixed to paramagnetic beads, which then were exposed to a magnetic field and sorted. X‐ and Y‐chromosome frequencies in the two subgroups of sperm were assayed by multiprobe fluorescent in situ hybridization (FISH).

RESULTS: Sperm were immunomagnetically separated into two populations: a reactive group (presumably, H‐Y Ag+); and a nonreactive group (presumably, H‐Y Ag‐). Triple‐color FISH analysis of 1,600 spermatozoa (800 in each group) showed the antigen's expression to be somewhat more prevalent among Y‐chromosome‐bearing sperm (54.1%), but a large proportion of Y‐chromosome‐bearing sperm (49.0%) did not express this antigen. The difference was not significant (P = 0.43).

CONCLUSIONS: The expression of H‐Y antigen has a slightly higher frequency in human sperm containing the Y‐chromosome, but its expression among X‐chromosome‐bearing sperm also is considerable. Current immunologic techniques relying on this antigen are unlikely to effect the sex selection of human sperm.
Secretion of Interleukin‐2 from Unstimulated Peripheral Blood Mononuclear Cells Is a Possible Pathogenic Mechanism in Recurrent Abortion
Authors: HAMAI YOKO , FUJII TOMOYUKI , KOZUMA SHIRO , SHIBATA YOICHI , TAKETANI YUJI
Published: September 2011

PROBLEM: The objective of this study was to evaluate the activated status of helper T cells in women with recurrent abortion.

METHOD OF STUDY: The spontaneous secretion of interleukin (IL)‐2 from peripheral blood mononuclear cells (PBMCs) obtained from 12 women with primary partner‐specific recurrent abortion and 7 normal females and 10 normal males was measured.

RESULTS: IL‐2 concentrations in medium conditioned with PBMCs from the recurrent abortion group were 1.44 ± 0.32 (mean ± standard deviation) U/ml, whereas those from the normal female and male groups were below the minimum detectable concentration.

CONCLUSION: The fact that PBMCs from recurrent aborters spontaneously secrete IL‐2 suggests that helper T cells are activated in recurrent aborters.
Levels of Antibodies to Transferrin and Alpha 2‐HS Glycoprotein in Women With and Without Endometriosis
Authors: Mathur Subbi Pillai , Rust Philip F. , Holt Victoria L. , Jiang Haixiang , Lee Jong H.
Published: September 2011

PROBLEM: To establish an enzyme‐linked immunosorbent assay (ELISA) for determining the levels of antibodies to transferrin and alpha 2‐HS glycoprotein in the serum of women with and without endometriosis.

METHOD OF STUDY: Serum samples were obtained from 105 normal women, who were randomly selected for a population‐based epidemiologic study, and 123 patients with active endometriosis. An ELISA using transferrin and alpha 2‐HS glycoprotein as antigens was established.

RESULTS: The levels of antibodies to transferrin and alpha 2‐HS glycoprotein in the serum of patients with endometriosis were approximately 21 times higher than those in the serum of control subjects without endometriosis. Only 2% of control subjects had false positive levels of these antibodies, and 5% of patients with endometriosis had false negative levels of these antibodies (specificity, 98.1 and 98.1, respectively, for anti‐transferrin and anti‐alpha 2‐HS glycoprotein; sensitivity, 95 and 96.7, respectively, for anti‐transferrin and anti‐alpha 2‐HS glycoprotein).

CONCLUSIONS: Patients with endometriosis have significantly higher levels of antibodies to transferrin and alpha 2‐HS glycoprotein than control subjects. Testing women for antibodies to transferrin and alpha 2‐HS glycoprotein will provide a specific noninvasive diagnosis of endometriosis.
Fertility‐Disrupting Potential of Synthetic Peptides Derived from the β‐Subunit of Follicle‐Stimulating Hormone
Authors: FERRO VALERIE A. , STIMSON WILLIAM H.
Published: September 2011

PROBLEM: Hormone immunoneutralization is hampered by immunologic cross‐reactivity caused by close‐sequence homology between related molecules. One solution is to use smaller fragments to induce antibodies of greater specificity.

METHOD OF STUDY: A number of peptides selected from β‐follicle‐stimulating hormone (FSH) were conjugated to tetanus toxoid and were used to immunize female rats. The antis‐era were examined for FSH cross‐reactivity by immunoassays and in an in vitro bioassay.

RESULTS: In the immunoassays, the antisera did not react with FSH but did react with their respective peptides. In the bioassay, sera from VYKDPARPC‐ and CDSLYTYP‐immunized animals inhibited FSH‐receptor interaction by 73% and 68%, respectively. These animals also showed reduced estradiol levels. Sequences were synthesized around VYKDPARPC and were tested on a FSH‐receptor‐bearing Chinese hamster ovary cell line. LVYKDPARPC, VYKDPARPC, YKDPARPIC, CLVYKDPARP, and LVYKDPARP inhibited FSH‐receptor interaction by greater than 50%. In female mice, TRDLVYKDPARPKI and LVYKDPARP disrupted estrous cycling in all animals; LVYKDPARPC and CLVYKDPARP disrupted cycling in three of five animals, whereas VYKDPARPC disrupted cycling in one of four animals.

CONCLUSIONS: Peptides from two areas of β‐FSH (VYKDPARP and DSLYTYP) were shown to raise FSH‐neutralizing antibodies, which were able to suppress estradiol levels. An additional leucine residue to VYKDPARP greatly enhanced the peptide's ability to inhibit FSH‐receptor binding and caused fertility disruption in vivo.
Evaluation of the Contraceptive Potential of Recombinant Human ZP3 and Human ZP3 Peptides in a Primate Model: Their Safety and Efficacy
Authors: PATERSON MARGARET , WILSON MARTIN R. , MORRIS KEITH D. , VAN DUIN MARCEL , AITKEN R. JOHN
Published: September 2011

PROBLEM: The unique recognition events that result in the avid binding of mammalian spermatozoa to the surface of the zona pellucida (ZP) are being exploited in the development of contraceptive vaccines. In this study, the safety and efficacy of a vaccination strategy based on the induction of active immunity against purified, glycosylated, recombinant human ZP3 (rhZP3) has been evaluated in a primate model, Callithrix jacchus.

METHOD OF STUDY: Long‐term infertility was established after immunization with rhZP3 and the resulting immune sera reacted with rhZP3 on an enzyme‐linked immunosorbent assay (ELISA) and immunolocalized exclusively to the outer surface of native ZP on marmoset ovarian sections. However, this contraceptive effect was inevitably associated with the eventual appearance of an ovarian pathology characterized by a depletion of primordial follicles. In an attempt to circumvent this side effect, human ZP3 (hZP3) was epitope mapped and four continuous, immunodominant B‐cell epitopes (hZP345–64, hZP393–110, hZP3172–190 and hZP3341–360) were evaluated for contraceptive efficacy in vivo. Using peptide‐tetanus toxoid (TT) conjugates to enhance immunogenicity, antipeptide antibodies were raised against these immunogens, which also cross‐reacted with rhZP3 on ELISA. In addition, antibodies against hZP345–64 and hZP3172–190 recognized native ZP on marmoset ovarian sections when a microwave technique was used to enhance epitope presentation.

RESULTS: No ovarian pathology was observed after the long‐term administration of these peptide immunogens, and fertility was suppressed when compared with TT controls but could not be correlated to the antibody titer.

CONCLUSION: Clearly, further research is required to identify optimal B‐cell epitopes that will reliably induce infertility, free from any ovarian pathology.
Increased Mast Cell Density in Peritoneal Endometriosis Compared with Eutopic Endometrium with Endometriosis
Authors: MATSUZAKI SACHIKO , CANIS MICHEL , DARCHA CLAUDE , FUKAYA TAKAO , YAJIMA AKIRA , BRUHAT MAURICE A.
Published: September 2011

PROBLEM: Factors regulating fibrosis in peritoneal endometriosis are poorly understood. We hypothesized that mast cell proteases may play a role in the process of fibrosis in peritoneal endometriosis. As the first step in examining the possible contribution of mast cells to fibrosis, we investigated their distribution in peritoneal endometriosis lesions according to their macroscopic appearance.

METHOD OF STUDY: Mast cells were identified by immunohistochemistry using monoclonal antibody against mast cell tryptase on formalin‐fixed, paraffin‐embedded sections. Mast cell density (mean mast cell count per 0.13 mm2 of stroma) was measured using a computerized image analysis system.

RESULTS: Mast cell density was significantly increased in black peritoneal lesions compared with red peritoneal lesions in endometriosis.

CONCLUSIONS: Mast cells may be involved in the pathogenesis of peritoneal endometriosis.
Effects of Pregnancy on Lymphocytes Within Sheep Uterine Interplacentomal Epithelium
Authors: FOX A. , LEE C.S. , BRANDON M.R. , MEEUSEN E.N.T.
Published: September 2011

PROBLEM: Previous studies demonstrate increases in the number and granularity of γδ T cells within the sheep uterine interplacentomal epithelium during pregnancy. To further characterize their activation and function, γδ T‐cell receptor (TCR)+ intraepithelial lymphocytes (IELs) from nonpregnant and pregnant uteri were phenotyped extensively. Cytokine mRNA expression in the epithelium and by γδTCR+ IELs isolated from pregnant uteri was also examined.

METHOD OF STUDY: Cell suspensions were prepared from the uterine interplacentomal epithelium and from the peripheral blood of nonpregnant and pregnant ewes (120–140 days of gestation). Surface marker expression was determined by two‐color flow cytometry and cytokine expression determined by reverse transcriptase‐polymerase chain reaction.

RESULTS: Uterine γδTCR+IELs exhibited increased β1‐integrin expression but decreased leukocyte function associated antigen (LFA)‐1 and major histocompatibility complex class I expression during pregnancy. Major histocompatibility complex class II, CD44, CD2, and LFA‐3 expression was unchanged during pregnancy, whereas CD25, VLA‐4 and L‐selectin were never expressed. The same cytokines were expressed in the pregnant and nonpregnant uterine interplacentomal epithelium with detectable mRNA for interferon (IFN)‐γ, tumor necrosis factor (TNF)‐α, and interleukin (IL)‐1α, but not for IL‐2 or IL‐4. γδTCR+ and CD8+ IEL purified from pregnant uteri expressed mRNA for IFN‐γ, TNF‐α, transforming growth factor‐β, and IL‐10.

CONCLUSIONS: γδTCR+ IELs from pregnant uteri have cytoplasmic granules, and express CD8 and cytokines indicative of cytotoxic potential. Phenotypic changes induced during pregnancy differed from those observed after activation of circulating naive cells and may represent further stimulation of fully differentiated effectors. γδTCR+ IELs are present only in interplacentomal areas of pregnant uteri and may control trophoblast invasion within these areas.