bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of Cultured Human Chorion Cell Chemokine Production by Group B Streptococci and Purified Bacterial Products
Authors: Dudley Donald J. , Edwin Samuel S. , Dangerfield Albina , Waggoner Jennifer Van , Mitchell Murray D.
Published: September 2011

PROBLEM: To determine if different strains of group B streptococci (GBS) and purified bacterial products regulate chemokine production by cultured human chorion cells.

METHOD OF STUDY: Primary cultures of human chorion cells were established from placentae isolated from normal women at term gestation having repeat cesarean section. Five different strains of heat‐killed GBS were incubated with confluent chorion cells at 107 bacteria/ml for 16 hours at 37°C. In separate experiments, lipoteichoic acid and sialic acid at various concentrations were incubated with chorion cells for 16 hours at 37°C. Culture supernatants were collected and then assayed to determine concentrations of interleukin‐8 (IL‐8) and macrophage inflammatory protein‐1α (MIP‐1α) by ELISA.

RESULTS: We found that GBS stimulated chorion cell production of MIP‐1α in a strain‐specific fashion. We also found that both lipoteichoic acid and sialic acid stimulated concentration‐dependent increases in chorion cell IL‐8 production. Chorion cells, however, did not increase MIP‐1α production in response to either lipoteichoic acid or sialic acid. Two strains of GBS tested induced concentration‐dependent increases in both IL‐8 and MIP‐1α, but both stimulated IL‐8 production to a greater extent. Similarly, IL‐1β also caused chorion cells to produce more IL‐8 than MIP‐1α.

CONCLUSIONS: Our data are the first to show that GBS and purified bacterial products can stimulate chemokine production by fetal gestational tissues. We suggest that chorion cells may produce specific types of chemokines to attract different types of inflammatory cells and thus may participate in the pathophysiology of infection‐mediated preterm labor by directing specific inflammatory responses.
Monoclonal Antibodies Reacting Against Capacitated but not With Freshly Ejaculated Boar Spermatozoa
Authors: Mollova Margarita V. , Atanasova Galina A. , Kyurkchiev Stanimir , Ivanova Maria D. , Peknicova Jana
Published: September 2011

PROBLEM: The involvement of individual sperm proteins in differentiation of antigenically specific and functionally defined regions on sperm membrane has not yet been completely elucidated.

METHOD: BALB/c mice were immunized with live capacitated boar spermatozoa and used for production of monoclonal antibodies (MAbs). ELISA, IIF, SDS‐PAGE, IVF, and cytologic methods were used for selection and biological characterization of MAbs as well as for identification of corresponding antigens.

RESULTS: MAb1F10, MAb2E2, and MAb4B12 react with antigens in the acrosome portion of live capacitated spermatozoa. MAb 1F10 reacted with human sperm cells along with those from bull, ram, mouse, dog, whereas MAb2E2—with mouse's spermatozoa and MAB4B12 ‐ with bull's, mouse's, and dog's spermatozoa. Some glycolytic enzymes seemed to reduce mildly the reactions of the MAbs with enzyme treated sperm cells; proteolytic enzymes eliminated the binding of MAbs to the sperm acrosome. These MAbs have no sperm agglutinating and/or sperm‐immobilizing activities and reduced the number of spermatozoa binding to zona pellucida.

CONCLUSIONS: MAb1F10, MAb2E2, and MAb4B12 seemed to recognize membrane associated antigens with potential role in the initial stages of fertilization, specific for capacitated but not for freshly ejaculated spermatozoa.
Altered Expression of Human Leukocyte Antigen G (HLA‐G) on Extravillous Trophoblasts in Preeclampsia: Immunohistological Demonstration With Anti‐HLA‐G Specific Antibody “87G” and Anti‐cytokeratin Antibody “CAM5.2”
Authors: Hara Naonori , Fujii Tomoyuki , Yamashita Takahiro , Kozuma Shiro , Okai Takashi , Taketani Yuji
Published: September 2011

PROBLEM: Human leukocyte antigen‐G (HLA‐G) is suggested to be at play in the materno‐fetal immune relationship during pregnancy. In the light of current concept that disruption of the materno‐fetal immune relationship could account for several complications of pregnancy, including preeclampsia, we asked whether the expression of HLA‐G protein on the trophoblasts is altered in preeclampsia.

METHOD: The presence of HLA‐G protein in the extravillous trophoblasts in placenta obtained from five preeclamptic patients and seven uncomplicated pregnant women was determined by means of an immunohistochemical technique.

RESULTS: All of the extravillous trophoblasts, which were stained for cytokeratin, were stained for HLA‐G protein in every woman with an uncomplicated pregnancy. In contrast, clusters of extravillous trophoblasts were insularly devoid of the staining for HLA‐G in all the preeclamptic patients.

CONCLUSION: The attenuated expression of HLA‐G protein on the extravillous trophoblasts could be at play in the pathophysiology of preeclampsia.
Placental Interferons
Authors: Bazer Fuller W. , Spencer Thomas E. , Ott Troy L.
Published: September 2011

Trophectoderm of ruminant conceptuses (embryo and associated membranes) secrete tau interferons (IFNτ) as the pregnancy recognition signal. Secretion of IFNτ on gestational days 12–13 for sheep and gestation days 14–17 for cows and goats is critical for pregnancy recognition. IFNτ acts on uterine epithelium to suppress estrogen receptor and oxytocin receptor gene expression, which prevents uterine release of luteolytic pulses of prostaglandin F2α (PGF). Expression of the progesterone receptor (PR) gene in uterine endometrium is not affected by oIFNτ. Maintenance of progesterone secretion by the corpus luteum (CL) ensures establishment of pregnancy. Pig conceptuses secrete both IFNα and IFNγ between days 15–21 of gestation, but their role(s) in early pregnancy is unknown. Estrogen secreted by pig trophoblast between gestational days 11–13 and 15–25 increases endometrial receptors for prolactin and causes exocrine secretion of PGF into the uterine lumen to prevent luteolysis. Shared cell‐signaling mechanisms by IFNs and lactogenic hormones through Janus kinases (JAK) 1 and 2 may provide a common pathway to abrogate luteolytic mechanisms to ensure establishment of pregnancy. The role(s) of IFNs produced by human and rodent placentae is not known.
The Immunological Pregnancy Protective Effect of Progesterone Is Manifested via Controlling Cytokine Production
Authors: Szekeres‐Bartho Julia , Faust Zs. , Varga P. , Szereday L. , Kelemen K.
Published: September 2011

PROBLEM: This study was aimed at investigating the involvement of an altered cytokine pattern in the immunomodulatory and anti‐abortive effects of a progesterone‐induced immunomodulatory protein (PIBF).

METHOD: PIBF expression on lymphocytes of healthy pregnant women and from women at risk for premature pregnancy termination was determined. In sera of the same women TNFα was quantified by a bioassay using L929 cells. NK activity was determined by a single cell cytotoxicity assay. Cytokine production of the lymphocytes or murine spleen cells was measured by ELISA or detected by immunocytochemistry. In pregnant mice endogenous PIBF activity was neutralized by anti‐PIBF IgG.

RESULTS: Sera of women at risk for premature pregnancy termination contained significantly higher concentrations of TNFα than those from healthy pregnant women and PIBF expression on the lymphocytes was inversely related to serum concentration of TNFα. Increased NK activity of lymphocytes after neutralization of endogenous PIBF activity is corrected by anti‐IL2 treatment and PIBF inhibits IL12 expression on activated lymphocytes. PIBF increases IL‐10 production by activated spleen cells. In pregnant mice, neutralization of endogenous PIBF activity by specific antibody results in increased resorption rate and reduced splenic IL‐10 production.

CONCLUSIONS: Our data allow the assumption that via blocking IL‐12 production PIBF inhibits NK activation with a concomitant reduction of TNFα levels. Disturbances in this system might lead to the expression of the known synergistic effect of IL‐12 and TNFα, resulting in a Th1 type cytokine dominance and pregnancy termination.
Elevated Peripheral Blood Natural Killer Cells Are Effectively Downregulated by Immunoglobulin G Infusion in Women With Recurrent Spontaneous Abortions
Authors: Kwak J.Y.H. , Kwak F.M.Y. , Ainbinder S.W. , Ruiz A.M. , Beer A.E.
Published: September 2011

PROBLEM: We investigated the hypothesis that elevated peripheral blood natural killer cells (NK) are decreased by immunoglobulin G infusion (IVIg) therapy in women with recurrent spontaneous abortions (RSA) and elevated NK cells.

METHODS: Seventy‐three women with RSA and elevated NK cells received IVIg therapy (400 mg/Kg/day for 3 days ever 4 wks) and anticoagulation treatment. Peripheral blood immunophenotype assay by flow cytometry was done prospectively prior to and 7 days after first IVIg therapy, every 2 wks until 20 wks gestation and then monthly. Controls were 95 women with RSA and normal NK cells who received anticoagulation treatment.

RESULTS: (1) 86.3% of women with elevated NK cells who received the IVIg and anticoagulation therapy had a successful pregnancy outcome; (2) Peripheral blood CD56+ NK cells and CD56+/16+ NK cells were significantly suppressed 7 days post IVIg infusion (P < 0.0005); (3) Pre‐IVIg infusion levels of other lymphocyte subsets were not different as compared with those of 7 days post‐IVIg therapy; (4) Women who delivered a liveborn infant with IVIg therapy demonstrated downregulation of peripheral blood NK cells (CD56+, CD56+/16+) during early pregnancy when compared to women who miscarried the index pregnancy (P < 0.05); (5) Women with normal NK cells who miscarried while on anticoagulation therapy demonstrated significantly elevated CD56+ NK cells during early pregnancy as compared with that of women who delivered a liveborn infant (P < 0.05); (6) CD19+ B cells were significantly downregulated during pregnancy in women with anticoagulation and IVIg therapy when compared to women with anticoagulation therapy (P < 0.05).

CONCLUSION: Downregulation of NK cells in women with RSA is associated with a favorable pregnancy outcome. Peripheral blood NK cells (CD56+, CD56+/16+) are effectively suppressed after IVIg therapy. Women with RSA and high NK cells benefit from IVIg therapy and experience suppression of CD56+ and CD56+/16+ NK cells.
Laboratory Evaluation of Women Experiencing Reproductive Failure
Authors: Roussev Roumen G. , Kaider Brian D. , Price David E. , Coulam Carolyn B.
Published: September 2011

Reproductive life table analysis indicates that the majority of reproductive failures result from post fertilization failures, whether before or after implantation. It is important to have a set of tests to clarify the diagnosis of the reproductive failure so that appropriate therapy can be instituted. To determine the frequency of abnormal immunologic tests among women experiencing reproductive failure, 108 patients were evaluated for the presence of antiphospholipid antibodies (APA); lupus anticoagulant (LA); thyroid‐thyroglobulin and microsomal antibodies (TGT); embryotoxic factor (ETA); and systemic CD56+/CD16‐ cells. The frequency of abnormal results obtained from testing for APA, LA, TGT, ETA, and CD56+/CD16‐ cells among 108 patients with diagnoses of recurrent pregnancy loss (RPL)(n=45), unexplained infertility (n=45) including IVF failure (n=10), endometriosis (n=10), premature ovarian failure (n=5), and polycystic ovaries (n=3) were compared with 15 normal controls. Seventy of one hundred eight (65%) women experiencing reproductive failure had at least one positive test, compared to 1 of 15 (7%) controls (P=0.0001). Presence of phospholipid antibodies was the most frequently abnormal result followed by elevated CD56+/CD16 cells. The prevalence of a particular abnormal test varied among the diagnoses. The most frequent abnormal test among women with RPL was an increased percentage of CD56+/CD16‐ cells (40%), followed by APAs (29%), TGT (9%), and ETA (7%). The most frequent abnormal result among women with unexplained infertility was the presence of APAs (42%), followed by CD56+/CD16‐ cells (16%), ETA (16%), and TGT (9%). APA, CD56+/CD16‐ cells, ETA, and TGT are useful tools to assist in the diagnosis of reproductive failure.
Cervical Mucus Anti‐Sperm Antibodies: Treatment With Intrauterine Insemination
Authors: Kutteh William H. , Byrd William , Blankenship Linda , Kutteh Carol C. , Carr Bruce R.
Published: September 2011

PROBLEM: To determine (1) the incidence of cervical mucus anti‐sperm antibodies in infertile women, and (2) the results of treatment by intrauterine insemination.

METHOD: Cervical mucus was collected the morning after urinary LH surge occurred from 153 consecutive women being treated for unexplained infertility with intrauterine insemination. Immunobead testing for IgG, IgA, IgA1, and IgA2 was performed with only actively motile sperm being counted.

RESULTS: Overall, 23/153 (15.0%) of cervical mucus samples were positive for anti‐sperm antibodies: 9/23 (39.1%) were only IgA‐positive (62% IgA1‐positive, 38% IgA2‐positive), 11/23 (47.8%) were only IgG‐positive, and 3/23 (13.0%) were positive for both IgA and IgG. Insemination resulted in a pregnancy in 6/23 (26.1%) of women with cervical mucus anti‐sperm antibodies after 1–3 cycles.

CONCLUSIONS: Testing for cervical mucus anti‐sperm antibodies should be performed in cases of “unexplained” infertility, and intrauterine insemination may be an effective treatment, resulting in pregnancies in over one‐fourth of couples.
Genetic Factors in Reproduction and Their Evolutionary Significance
Authors: Gill Thomas J.
Published: September 2011

PROBLEM: The reproductive process is a major driving force in human evolution. An evolutionary perspective was brought to bear on some aspects of reproduction and its aberrations, and, conversely, some of the insights of modern reproductive genetics were used to investigate problems in evolution.

METHOD: The data used were obtained from the literature in evolution, anthropology, archeology, linguistics, and genetics.

RESULTS: The evolutionary line leading to modern humans diverged from that leading to the chimpanzees approximately 5–7 million years ago (Mya). Archaic Homo sapiens emerged ca. 0.3 Mya, and modern Homo sapiens and the development of language ca. 0.1 Mya; thus, modern humans occupy approximately 2% of the evolutionary history of the hominid line. During all of this time, the ancestors of modern humans were migratory hunter‐gatherers. It was only during the Neolithic transition ca. 0.01 Mya (approximately 0.2% of of hominid evolutionary history) that agriculture was developed, and with it a settled lifestyle that allowed a more stable existence and the development of a different reproductive pattern. Various estimates indicate that the human population increased from 0.05 million at the time of the emergence of modern Homo sapiens to 6,000 million at the present time (120,000‐fold increase).

CONCLUSIONS: These evolutionary considerations were used to explore three areas: (1) the extinction of the Neanderthals, who coexisted for ca. 65,000 years with modern humans; (2) the relatively low and stable rate of human conceptions (20–35% of ova fertilized naturally or fertilized in vitro); and (3) the long postnatal period required for the full maturation of the immune response. From these considerations, a broad view of the human reproductive process was obtained that may provide some insight into the rationale for the development of effective reproductive technologies.
Lymphocyte Immunotherapy (LI) Increases Serum Levels of Progesterone Induced Blocking Factor (PIBF)
Authors: Check Jerome H. , Arwitz Maria , Gross Jenifer , Peymer Mark , Szekeres‐Bartho Julia
Published: September 2011

PROBLEM: To determine if allogenic stimulation from leukocyte immunization (LI) can increase the production of an immunomodulatory protein called progesterone induced blocking factor (PIBF) by CD8+ T‐lymphocytes.

METHOD: The study group consisted of 35 women, 29 who failed to conceive after repeated embryo transfers (ETs) and six with recurrent spontaneous abortion (RSA). The women underwent LI using the male partner's blood as the source of leukocytes. Progesterone induced blocking factor was measured pre‐ and post‐LI with an immunocytochemistry method using a PIBF‐specific polyclonal antibody.

RESULTS: The mean percentage of lymphocytes expressing PIBF, as well as the percentage of cases whose PIBF level increased to 1% or more, was significantly higher post‐LI. Similarly post‐LI, there was a significantly lower percentage of zero PIBF levels.

CONCLUSIONS: Leukocyte immunization causes an increase in PIBF in many cases. Possibly the improved pregnancy outcome in immunized patients with RSA or previous failure to conceive with in vitro fertilization may be partially or possibly completely explained by its stimulatory effect on PIBF.
The Implications of a Paternally Derived Centrosome During Human Fertilization: Consequences for Reproduction and the Treatment of Male Factor Infertility
Authors: Navara Christopher S. , Hewitson Laura C. , Simerly Calvin R. , Sutovsky Peter , Schatten Gerald
Published: September 2011

PROBLEM: Successful fertilization in humans follows a complex series of events, including the completion of meiotic maturation of the oocyte with the extrusion of the second polar body, the decondensation of the sperm nucleus and the maternal chromosomes into male and female pronuclei, the restoration of the sperm centrosome, and the nucleation of microtubule‐mediated motility necessary to bring the male and female pronuclei into close apposition. These events occur after both fertilization in vitro and after intracytoplasmic sperm injection (ICSI), a new technique which is currently being applied in many clinics to overcome severe male infertility. Defects in any of the events leading to fertilization can be lethal to the zygote and may prove to be causes of infertility.

METHODS: Imaging of inseminated human and rhesus oocytes using immunohistochemical techniques reveals several phases at which fertilization arrests.

RESULTS: Oocytes from some infertile patients failed to complete fertilization due to failure of the sperm aster microtubules in uniting the sperm and egg nuclei. The rate of sperm aster formation, size, and organization during fertilization has been used as a measurement of bovine sperm quality. The development of an assay using Xenopus laevis oocyte extract can also be used to test sperm from various species for their ability to form asters and perform other centrosomal functions in vitro, as well as another indicator of sperm quality. Semen from men with questionable fertility was found to contain sperm which are generally incapable of producing sperm asters. In addition, the activity of centrosomal proteins such as γ‐tubulin and centrin have been detected in mammalian eggs and sperm. The levels of γ‐tubulin increase markedly after exposure to X. laevis egg extract.

CONCLUSION: Defects in either male or female nucleus decondensation also resulted in the arrest of fertilization and was found to occur in both inseminated human oocytes and in rhesus oocytes fertilized by ICSI. These discoveries on the molecular basis of infertility in humans have important implications for infertility diagnosis and managing reproduction.
Review: Cytokine Involvement in Ovarian Processes
Authors: Terranova P.F. , Rice V. Montgomery
Published: September 2011

PROBLEM: Expression of tumor necrosis factor‐α (TNF) and interleukins 1α and 1β (IL‐1) have been reported in ovaries of several species and humans and are implicated in ovarian follicular development and atresia, ovulation, steroidogenesis, and corpus luteum function (including formation, development, and regression). The principal abnormal processes affected by these cytokines are ovarian cancer and reduction of ovarian function during sepsis.

METHODS: A literature review.

RESULTS: Numerous studies indicate that TNF and IL‐1 inhibit gonadotropin‐stimulated steroidogenesis of undifferentiated ovarian cells due to inhibition of adenylyl cyclase and post‐cAMP sites. In differentiated ovarian cells, these cytokines either stimulate progesterone synthesis or have little to no effect on steroidogenesis. Both cytokines participate in ovulation and levels of these cytokines increase during the periovulatory period. Endotoxin inhibits gonadotropin‐stimulated ovarian steroidogenesis and follicular development and these effects are mediated, in part, by TNF and by direct effects of endotoxin on ovarian cells. In newly formed corpora lutea, progesterone secretion is inhibited by TNF and IL‐1, although each has proliferative effects. TNF also has been implicated in regression of corpora lutea because TNF stimulates prostaglandin synthesis and luteal TNF increases after initiation of the decline in progesterone secretion. TNF and IL‐1 are secreted by some ovarian cancer cells and stimulate growth of these cells.

CONCLUSIONS: Thus, TNF and IL‐1 are multifunctional factors affecting various ovarian processes.
Stress Triggered Abortions Are Associated With Alterations of Granulated Cells in the Decidua
Authors: Markert Udo R. , Arck Petra C. , McBey Betty Ann , Manuel Justin , Croy B. Anne , Marshall Jean S. , Chaouat Gerard , Clark David A.
Published: September 2011

PROBLEM: Stress is known to be abortogenic in animals and humans. An increased decidual release of cytokines such as TNF‐α and reduction in TGF‐β2‐related immunosuppressive activity has been proposed as the triggering mechanism. Substance P released by nerves in endometrium/decidua has been found to be the key neurotransmitter in this pathway. It is still unclear which cells are stimulated by substance P to produce the increased TNF‐α level.

METHOD: As a measure of local activation, the granulation of granulated metrial gland (GMG) cells was measured by flow cytometry after sonic plus immobilization stress of mice or substance P treatment of GMG cells (both isolated GMG cells and GMG‐cell containing decidua). TNF‐α release from decidua and isolated GMG cells was investigated using a TNF‐α bioassay. The degranulation of uterine mast cell, another potential source of TNF‐α, was examined in situ by Toluidine blue staining.

RESULTS: We observed a striking increase in percentage of degranulated mast cells (8% r̊ 24%) in the uteri of stressed animals, whereas the granularity of GMG cells was decreased by stress but increased with treatment with substance P in vitro. Isolated GMG cells appeared to release in vitro cytotoxins active in the TNF‐α bioassay, but the magnitude of this activity was not increased by stress or by substance P treatment. In contrast, disaggregated decidual tissue which is known to release increased amounts of TNF‐α after stress, did increase activity in response to substance P in vitro.

CONCLUSIONS: Uterine mast cells show activation as reflected by degranulation after stress exposure of pregnant mice and mast cells might be the cellular link between the neurotransmitter substance P and an increase in decidual TNF‐α release that leads to abortion.
Induction of Experimental Antiphospholipid Antibody Syndrome in PL/J Mice Following Immunization With β2GPI
Authors: García César O. , Kanbour‐Shakir A. , Tang Hua , Molina José F. , Espinoza Luis R. , Gharavi Azzudin E.
Published: September 2011

PROBLEM: Immunization with β2‐glycoprotein I (β2GPI) induces antiphospholipid antibodies (aPL) in normal mice and rabbits. Recently we reported early onset of autoimmunity in MRL/++ mice following immunization with β2GPI. There is a close association between aPL with thrombosis, recurrent fetal loss, and intrauterine growth retardation. In this study we evaluated the effect of β2GPI‐induced aPL on pregnancy outcomes in an inbred strain of mice (PL/J).

METHOD: Three groups of seven‐week‐old female PL/J mice (12 per group) were studied. Group A was immunized with β2GPI and group B with ovalbumin; group C was not immunized. After two booster injections, the mice were tested for aPL, anti‐DNA by ELISA, and for ANA by indirect immunofluorescence. Platelet count and pregnancy outcomes were studied at the age of 14 weeks.

RESULTS: The aPL and anti‐DNA levels were higher at 12 and 14 weeks in group A; the optical densities (OD) were 1.72±0.6 and 0.699±0.25 for group A, 0.091 ±0.040 and 0.230±0.47 for group B, and 0.0435±0.003 and 0.119±0.026 for group C (comparing group A with groups B and C combined, P<0.001). ANA titers rose in groups A and B by age, but they were significantly higher at 14 weeks in group A. The mean titers were 1/286, 1/90, and 1/16 for A, B, and C, respectively (P<0.001). The platelet counts were not significantly different among the three groups. The litter size was significantly smaller in group A, as evidenced by the numbers of viable fetuses among the mice that became pregnant in each group: 0.75, 2.45, and 5.5 in groups A, B, and C, respectively. Seven pregnant mice in group A had complete resorption, seven pregnant mice in group B showed focal (partial) resorption areas, and only one mouse in group C had complete resorption of the embryos, as shown by histopathological studies, although the fecundity rate was similar in the three groups.

CONCLUSION: Our data suggest a pathogenic role for β2GPI‐induced aPL in the development of experimental models of APS in PL/J mice.
TGFβ2 in Rabbit Blastocoelic Fluid Regulates CD4 Membrane Expression: Possible Role in the Success of Gestation
Authors: Ouellette Marie‐Josée , Dubois Claire M. , Bergeron Dominic , Roy Raynald , Lambert Raymond D.
Published: September 2011

PROBLEM: During pregnancy, major changes occur in the decidual cell population. One of these changes involves some phenotypical transformations of lymphocyte sub‐populations. Since these variations might be due to the presence of the embryo, the current study was designed to investigate the implication of blastocoelic fluid (BF) in these changes and to determine the mechanism by which this phenomenon occurs.

METHOD: Lymphocytes isolated from human peripheral blood (PBL) were cultured for 72 h in RPMI‐FCS 10% and with or without BF day 12 (BF d‐12) or Concanavalin A (ConA). After 72 h, T cells were labelled with anti‐CD4 antibodies and Quantum Simply Cellular microbeads were used as a standard to evaluate the antibody binding capacity (ABC).

RESULTS: Treatment of human PBL with BF d‐12 decreases the percentage of CD4 and TCR positive cells, as compared to non‐stimulated cells, but has no significant effect on CD2, CD3, and CD8 positive cells. It was also demonstrated, for the first time, that transforming growth factor beta‐2 (TGFβ2) in BF day 12 diminishes the percentage of CD4 positive cells by downregulating CD4 membrane expression on leucocytes.

CONCLUSION: These findings suggest that the embryo plays a key role in its own protection. Furthermore, it is predicted that any tissue producing TGFβ2, such as certain types of tumor, downregulates the immune response, thus allowing tumor growth.
Expression of CD15 (Lewisx) Antigen on Human Sperm and its Role in Sperm‐Egg Interaction
Authors: D'Cruz Osmond J. , Lambert Hovey , Haas Gilbert G.
Published: September 2011

PROBLEM: The carbohydrate epitope 3‐fucosyl‐N‐acetyllactosamine (CD15) is a constituent of cell surface glycoconjugates that has been implicated in cell‐cell adhesion mediated by carbohydrate‐specific ligands. The present study was designed to investigate whether CD15 is present on human sperm and whether it plays a role in human sperm‐egg interaction.

METHODS: Fluorescent flow cytometry was used to quantitate the binding of monoclonal antibodies (mAb) to sperm‐bound CD15 and CD46 antigens on acrosome‐intact (AI) and acrosome‐reacted (AR) sperm. The location of the binding site of these mAbs was assessed by fluorescence microscopy. The effects of anti‐CD15 and anti‐CD46 mAbs on gamete interaction were tested utilizing both homologous (human zona binding and penetration) and heterologous (zona‐free hamster egg binding and penetration) systems.

RESULTS: The mean percentage of capacitated sperm which bound anti‐CD15 or anti‐CD46 mAbs was low (4.8% and 5.1%, respectively). Exposure to calcium ionophore A23187 (CaI) resulted in an increase in anti‐CD15 (38.6 ± 4%) and anti‐CD46 (83.4 ± 2%) binding to sperm. Both anti‐CD 15 and anti‐CD46 binding sites were localized by fluorescence microscopy on the sperm acrosomal region. In four experiments, the percent of zona‐free hamster eggs penetrated by human sperm were medium control 93% (62/66), irrelevant mAb 74% (70/94), anti‐CD46 0% (0/107), and anti‐CD15 10% (9/90). One hundred percent (6/6) of human zona were penetrated by human sperm exposed to medium control, 88% (8/9) following exposure to irrelevant mAb, 0% (0/11) following exposure to anti‐CD46, and 50% (5/10) following exposure to anti‐CD 15. The mean (± SD of tightly bound sperm to hamster eggs were medium control 57 ± 18%, irrelevant mAb 64 ± 16%, anti‐CD46 37 ± 13%, and anti‐CD15 19 ± 10%. The corresponding values for human zona were: medium control 118 ± 14%, irrelevant mAb 61 ± 11%, anti‐CD46 39 ± 18%, and anti‐CD15 99 ± 19%.

CONCLUSION: CD15 antigen is expressed on human sperm that have undergone acrosomal loss. mAb to CD15 was shown to inhibit significantly sperm binding and penetration of zona‐free hamster eggs and penetration of human zona pellucida. These findings suggested that sperm‐egg interaction may be mediated in part by the CD15 antigen.

Capsule: Acrosome‐reacted human sperm bind monoclonal antibodies specific for CD15 (Lewisx) epitope. The binding sites were located on the sperm head. Anti‐CD15 antibody impaired both the binding and penetration of zona‐free hamster eggs and the penetration of human zona by human sperm.
Interleukin‐10 in Preovulatory Follicular Fluid of Patients Undergoing In‐Vitro Fertilization and Embryo Transfer
Authors: Geva Eli , Lessing Joseph B. , Lerner‐Geva Liat , Azem Foad , Yovel Israel , Ben‐Yosef Dalit , Barkai Uriel , Amit Ami
Published: September 2011

PROBLEM: The present study aimed at investigating the presence of Interleukin‐10 (IL‐10) in preovulatory follicular fluid (FF) and its possible correlation with 17‐β‐estradiol (E2) and progesterone (P) levels, and treatment outcome in patients undergoing in‐vitro fertilization‐embryo transfer (IVF‐ET).

METHODS: Twenty consecutive patients with tubal factor infertility who underwent oocyte retrieval for IVF‐ET were assayed for pooled, preovulatory FF levels of IL‐10, E2, and P.

RESULTS: The mean FF levels of IL‐10, E2, and P were 78.7 ± 104.7 pg/ml, 2,787.0 ± 726.1 pg/ml, and 1.5 ± 0.8 ng/ml, respectively. No correlation was found between preovulatory FF concentration of IL‐10, E2, oocyte number, oocyte fertilization rate, embryo quality, and pregnancy rate. The levels of IL‐10 were found to be negatively correlated with P concentration, although not significantly (P = 0.057).

CONCLUSION: Interleukin‐10 exists in the preovulatory FF. Further investigations are needed to determine the role of IL‐10 in the folliculogenesis.
Determination of IL‐1 and IL‐6 Levels in Human Embryo Culture‐Conditioned Media
Authors: Barañao R.I. , Piazza A. , Rumi L.S. , Fried E. Polak de
Published: September 2011

PROBLEM: The aim of this study was to evaluate the presence of interleukin‐1 (IL‐1) and IL‐6 in 24‐hour human embryo culture‐conditioned media (HECCM) and to find any embryo‐related factor(s) to predict pregnancy during IVF procedure.

METHOD: IL‐1β and IL‐6 levels were measured in 36 samples of HECCM and 17 cell culture media that had not been exposed to embryos, which were used as controls. Both cytokine levels were measured by the ELISA technique, using commercially available kits.

RESULTS AND CONCLUSIONS: We found an average IL‐1β level ± SEM of 82 ± 4 pg/ ml (n=11) in HECCM from viable pregnancies and a significantly lower value, 14 ± 2 pg/ ml (n=25, P<0,001), in HECCM from embryos that did not lead to viable pregnancies. In control media the average IL‐1β level was 11 ± 1 pg/ml (n=17), (P <0.001 vs. HECCM from viable pregnancies). In contrast IL‐6 levels were undetectable in all samples analyzed. Judging by our results we suggest that measurement of IL‐1 level in HECCM could be a useful parameter for predicting implantation in techniques of assisted reproduction.
Modulations of Cytokine Expression in Pregnant Women
Authors: TRANCHOT‐DIALLO JULIETTE , GRAS GABRIEL , BENVENISTE OLIVIER , MARCÉ DOMINIQUE , ROQUES PIERRE , DORMONT DOMINIQUE , PARNET‐MATHIEU FRANÇOISE , MILLIEZ JACQUES , CHAOUAT GÉRARD
Published: September 2011

PROBLEM: Although the overall anti‐infectious and anti‐parasitic immunity of parous women appears normal, several aspects of maternal cell‐mediated and humoral immunity are altered during pregnancy. This has been suggested to occur via preferential local and systemic secretion of Th‐2 type cytokines, which down‐regulate or prevent secretion/action of Th‐1 type cytokines, in animals as well as in humans.

METHOD: To evaluate the influence of gestation on the maternal immune system, we have measured, in pregnant women, the mRNAs coding for several cytokines (TNFα, IFNγ, GM‐CSF, IL‐Iβ, IL‐2, IL‐4, IL‐6, and IL‐10) in peripheral blood mononuclear cells, by use of semi‐quantitative RT‐PCR.

RESULTS: Our results show significant modulations of IFNγ, IL‐1β, IL‐4, and IL‐6 genes expression especially during the third trimester and near parturition. CONCLUSION: Cytokine expression is thus finely tuned in peripheral blood during pregnancy, in a previously unexpected complex pattern, related to gestational stage.
Midtrimester Amniotic Fluid Tumor Necrosis Factor‐Alpha Does Not Predict Small‐for‐Gestational‐Age Infants
Authors: SPONG CATHERINE Y. , SHERER DAVID M. , GHIDINI ALESSANDRO , PEZZULLO JOHN C. , SALAFIA CAROLYN M. , EGLINTON GARY S. , SPONG CATHERINE Y. , SHERER DAVID M. , GHIDINI ALESSANDRO , PEZZULLO JOHN C. , EGLINTON GARY S. , SALAFIA CAROLYN M.
Published: September 2011

PROBLEM: To evaluate the independent ability of midtrimester amniotic fluid tumor necrosis factor‐alpha (TNF‐α) in the prediction of small‐for‐gestational‐age (SGA) infants.

METHOD OF STUDY: In this case‐control study, patients delivering a SGA infant were matched with controls based on GA at delivery, maternal age, race, and parity. Patients with immune disease, chronic hypertension, diabetes, asthma, congenital hearts disease, multiple gestation, and fetal anomalies were excluded. Amniotic fluid samples were immunoassayed for TNF‐α. Potential confounding variables evaluated were maternal serum alpha‐fetoprotein level, smoking history, pregnancy induced hypertension, and neonatal gender. Statistical analysis included Fisher's exact test and ANOVA after log transformation with P < 0.05 considered significant.

RESULTS: Eighteen patients delivered SGA neonates and were matched with 41 controls. No significant differences were identified in the confounding variables between patients with SGA neonates and controls. Amniotic fluid TNF‐α levels were not significantly different between patients subsequently delivering SGA neonates and controls [median 7.63 (range 0.25‐16.1) pg/mL versus 9.39 (0.25–66.9) pg/mL, P = 0.8].

CONCLUSIONS: Midtrimester amniotic fluid TNF‐α levels are not predictive of SGA neonates when compared with controls matched for gestational age at delivery.
Structure and Protective Capacity of Tetanus and Diphtheria Antibodies Produced During Human Pregnancy and Transferred to New‐Born
Authors: PASETTI MARCELA F. , DOKMETJIAN JOSÉ , ERIKSSON PATRICIA V. , MANGHI MARCELA A. , BRERO MARÍA LUISA , FERRERO FERNANDO
Published: September 2011

PROBLEM: The structure and protective activity of antibodies against tetanus (anti‐T) and diphtheria (anti‐D), produced during human pregnancy and transferred to new‐born, was studied.

METHOD: Antibody levels were measured by ELISA in non‐pregnant women (control group), primiparae, and multiparae, and in their children. The proportion of symmetric and asymmetric IgG molecules was determined and their respective protective capacity evaluated.

RESULTS: The quantity of asymmetric anti‐T and anti‐D antibodies in mothers at the time of delivery was roughly four‐ and three‐fold that of the control group, respectively, dropping significantly 1 month later. A similar proportion of these antibodies was observed in the new‐born. The lower neutralizing capacity of asymmetric molecules was demonstrated in vivo.

CONCLUSION: Results show that during pregnancy there is a modulation of the immune response with an increase in the production of asymmetric molecules of lower protective capacity.
Soluble Receptors Neutralizing TNF‐α and IL‐1 Block Stress‐Triggered Murine Abortion
Authors: ARCK PETRA C. , CLARK DAVID A. , TROUTT ANTHONY B.
Published: September 2011

PROBLEM: In several models of abortion in rodents, the success or failure of the implanted embryos is determined by a balance between pro‐inflammatory cytokines such as tumor necrosis factor‐α (TNF‐α), interleukin‐2, interleukin‐1 (IL‐1), and γ‐interferon, and cytokines that counteract the former, such as interleukin‐10 and transforming growth factor‐β2 (TGF‐β2)‐related suppressor factor. Stress can trigger abortions in susceptible strains of mice and is thought to reflect the pathogenesis of some types of miscarriage in human pregnancy. In mice, stress increases levels of the abortogenic cytokine TNF‐α and decreases the suppressive activity of TGF‐β2‐related factor via a neurotransmitter substance P (SP)‐dependent pathway. Evidence for a role of pro‐inflammatory cytokines in SP‐mediated abortions in vivo is indirect.

METHODS: Direct evidence for a role of IL‐1 and TNF‐α in stress‐triggered abortions was sought by injecting pregnant female mice with soluble receptors neutralizing TNF‐α (rhuTNFR: Fc) or IL‐1 (rmIL‐1R) beginning 1 day after implantation and prior to stress.

RESULTS: The stress‐triggered abortion rate was reduced by 68% when either TNF‐α or IL‐1 antagonists were injected. The stress‐triggered decreased TGF‐β2‐like suppressive activity in the maternal uterine decidua was not restored by injection of either antagonist; indeed the soluble IL‐1 receptor significantly reduced suppressive activity in unstressed control mice, and soluble TNF‐α receptor had a lesser effect.

CONCLUSIONS: Both IL‐1 and TNF‐α play a role in the pathogenesis of stress‐triggered abortions, and may induce a compensatory physiological increase in suppressive activity in normal pregnancy counteracting pro‐inflammatory cytokines.
Comparison of the Immunobead Binding Test (IBT) and Immunospheres (IS) Assay for Detecting Serum Antisperm Antibodies
Authors: CENTOLA G.M. , ANDOLINA E. , DEUTSCH A.
Published: September 2011

PROBLEM: The IBT is considered the gold standard of sperm antibody assays. This test uses polyacrylamide beads labeled with antiglobulins (anti‐IgG, anti‐IgA, and anti‐IgM), which bind to the corresponding antibody on the sperm surface. The IS uses color‐coded latex beads of uniform 3.0 μm size coated with the antiglobulins which can be viewed with brightfield light microscopy. The purpose of the present study was to compare the IBT and IS in an indirect test using human serum.

METHOD: Serum specimens (n=42) were tested for the presence of antibody isotypes IgG, IgA, and IgM to sperm using the standard protocol for IBT and IS. Donor sperm was washed in BWW with 5% BSA and diluted to a final concentration of 50 times 106 motile sperm/ml. The sperm were incubated with a 1:10 dilution of test serum for 30 min to 1 h at 37°C and then washed by three cycles of centrifugation. The sperm and beads (IBT, IS) were mixed on a glass slide, covered with a coverslip, and observed within 5 min. At least 100 motile sperm were counted and scored for bead binding. A specimen was considered positive if 20% or more of the sperm were coated with one or more beads. The data were analyzed using calculation of the non‐parametric kappa statistic with correction for chance expected agreement, and by calculating the proportion of specific agreement between the two methods.

RESULTS: The results are summarized in the following table:

The IS was able to detect 94% of IgG antibodies, 91% of IgA antibodies, and 100% of Ig M antibodies. One serum specimen was IgG negative by IS (14% binding), but positive by IBT (20%). A second serum specimen was IgA negative by IS (16%) yet positive by IBT (29%). There were no false positives with the IS assay. Of the IgM positives (five of six) occurred alone and not with IgG or IgA, suggesting the necessity for testing all specimens also for IgM.

CONCLUSION: Antisperm antibody test results obtained by the IS assay are in agreement with the results obtained with the IBT test. The Immunospheres are monodispersed, color coded, and can be visualized with brightfield microscopy.
Efficacy of Intracytoplasmic Sperm Injection (ICSI) Treatment in the Immunological Infertile Patients
Authors: JUN JIN HYUN , LIM CHUN KYU , PARK YONG SEOG , LEE HO JOON , LEE YOU SIK , SEO JU TAE , SON IL PYO , KANG INN SOO
Published: September 2011

PROBLEM: To evaluate the efficacy of intracytoplasmic sperm injection (ICSI) treatment in the immunological infertile patients.

METHODS: The subjects were 60 cycles of 44 immunological infertile patients and these clinical data were analysed, retrospectively. They were classified into female and male immunological infertility group and were treated with conventional IVF, ICSI, and half‐ICSI.

RESULTS: The fertilization rate of ICSI (60.3%) and half‐ICSI (60.7%) cycles were significantly (P<0.01) higher than conventional IVF (42.6%) cycles. In the male immunological infertility group, fertilization rate was significantly (P<0.01) higher in ICSI (60.7%) and half‐ICSI (66.7%) cycles than in conventional IVF (27.4%) cycles and clinical pregnancy rate was higher in ICSI (54.5%) and half ICSI (33.3%) than in conventional IVF (25.0%) cycles.

CONCLUSIONS: We suggest that ICSI treatment is highly effective method to improve fertilization and pregnancy rate in IVF‐ET program of male immunological infertility.
Human Endometrial Leukemia Inhibitor Factor (LIF) Secretion and Its Relationship to Sonographic Endometrial Appearance
Authors: HAMBARTSOUMIAN EDOUARD
Published: September 2011

PROBLEM: LIF is believed to be involved in human reproduction. Because little is known about the function of this cytokine in proliferative phase of cycle and because LIF is found to regulate the cellular growth we evaluated the possible relationship between endometrial LIF secretion and endometrial growth.

METHOD: The present is prospective, blinded study with clinical and immunobiochemical correlation between endometrial LIF concentration and endometrial ultrasound pattern. Twenty‐four patients who were candidates for IVF and oocyte donation are included in this study. At day 10 of their cycle the endometrial biopsy was performed, followed by vaginal sonographic measurements of endometrial thickness and pattern. Endometrial LIF concentration was measured by ELISA in supernatants taken from cultured explants.

RESULTS: Endometrial LIF production significantly negatively correlated with endometrial thicknesses (P < 0.05). There was a 4‐fold elevation in LIF production when the endometrial thickness was below 5 mm. Strong correlation was found also between endometrial LIF production and sonographic endometrial pattern (P < 0.05). The most significant amount of LIF was found in nonfavorable endometria.

CONCLUSION: In the proliferative phase of cycle there is a dynamic relationship between endometrial sonographic appearance and local LIF secretion. Specifically LIF production is negatively correlated with endometrial thickness and pattern. The low amount of cytokine is a normal uteral environment for endometrial development, whereas deregulation of cytokine production toward its overexpression may lead to a strong inhibitory effect on endometrial growth. This finding might have an important clinical implication.
Major Goat Sperm 105 kDa Maturation Antigen: Purification, Characterization, and Effect of Its Antiserum on Acrosin Activity
Authors: Sarkar Madhumita , Das Tapasi , Chatterjee Tapati
Published: September 2011

A ConA binding membrane glycoantigen of 105 kDa molecular mass was purified from mature goat sperm by ion exchange and gel filtration chromatography. Of the detergents examined, the anionic deoxycholate was found to be highly effective in maximum solubilization of this sperm membrane antigen (SMA2). The analysis of the saccharide components by gas liquid chromatography revealed that the 105 kDa antigen (SMA2) contained the highest amount of mannose, followed by galactose and glucose in a ratio of 4:3:1. One amino sugar, N‐acetylglucosamine, was also found to be present in the polysaccharide branching of the SMA2 antigen. The internal sulfydryl linkage is essential for the maintenance of the protein backbone of 105 kDa antigen. The antigen selectively resides on the anterior head of goat sperm. The binding of anti‐SMA2 antibody to the integrated mature goat spermatozoa inhibited the release of acrosin after the induction of spermatozoa with Ca‐ionophore.
Interferon‐Gamma Production by the Human Preimplantation Embryo
Authors: ÖZörnek M. H. , Bielfeld P. , Krüssel J. S. , Copisti S. , Jeyendran R. S. , Koldovsky U.
Published: September 2011

PROBLEM: This study demonstrated that the human embryo produces interferon‐gamma (IFNγ). It is important to know whether IFNγ can be produced before implantation. Therefore the aim of this study was to evaluate the profile of IFNγ production between days 2 and 5 after in vitro fertilization.

METHOD: Twenty embryos were cultured from day 2 to 5 after fertilization. The embryo stages were checked each day and the media refreshed. IFNγ levels were estimated by an enzyme‐linked immunosorbent assay.

RESULTS: All embryos produced measurable IFNγ at least for 1 day. Yields of IFNγ were: 0.46 ± 0.45 (n = 4) on day 2, 0.69 ± 0.52 (n = 19) on day 3, 0.73 ± 0.52 (n = 15) on day 4, 0.55 ± 0.32 (n = 11) IU/ml on day 5, respectively. There was no significant difference in the IFNγ production between in vitro culture days or between the developmental stages of embryos.

CONCLUSION: IFNγ is produced by all the embryos and seems to peak between days 3 and 4, which is just before implantation.
Cytokine‐Leukocyte Networks and the Establishment of Pregnancy
Authors: Robertson Sarah A. , Mau Vicki J. , Hudson Sarah N. , Tremellen Kelton P.
Published: September 2011

PROBLEM: Factors in seminal plasma stimulate an intense but transient inflammatory response in the murine endometrium at mating. The aim of our current studies is to delineate the cytokine‐leukocyte interactions comprising this response and to elucidate the significance of these events in changes in the maternal immune system and as determinants of pregnancy outcome.

METHOD: We have reviewed our recent findings.

RESULTS: Transforming growth factor (TGF)‐β1 has been identified as the inflammation‐inducing moiety in seminal plasma. Seminal TGFβ1 initiates endometrial leukocyte infiltration by up‐regulating epithelial cell expression of granulocyte‐macrophage colony‐stimulating factor. Other cytokines and chemokines including regulated and normal T‐cell expressed and secreted (RANTES), macrophage inflammatory protein (MIP)‐1α, MIP‐1β, and monocyte chemotactic protein‐1 are also implicated as mediators of macrophage and granulocyte recruitment and activation. One consequence of this inflammatory response is the induction of a transient state of hyporesponsiveness to paternal major histocompatibility class I antigens.

CONCLUSION: Our studies suggest that semen may play a critical role in providing the antigenic and environmental signals necessary to initiate an appropriate maternal immune response to the conceptus during pregnancy.