bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of Interleukin (IL)‐6 and IL‐8 Production in an Amnion‐derived Cell Line by Cytokines, Growth Factors, Glucocorticoids, and Phorbol Esters
Authors: Keelan Jeffrey A. , Sato Timothy , Mitchell Murray D.
Published: September 2011

PROBLEM: To determine whether amnion cells produce interleukin (IL)−6 and −8 and thus may contribute to the high concentrations of these cytokines in amniotic fluid at term.

METHOD OF STUDY: Amnion‐derived WISH cells were treated in culture with stimuli over 16 hr, and IL‐6 and IL‐8 concentrations in the conditioned media were measured by enzyme‐linked immunosorbent assay or bioassay (IL‐6 only).

RESULTS: IL‐8 production was ˜5‐fold higher than that of IL‐6 under basal and stimulated conditions. Significant (by Dunnett's test after analysis of variance) stimulation of production of both cytokines was achieved by IL‐1β (>0.2 ng/ml), TNFα (>10 ng/ml), and the phorbol ester, phorbol 12‐myristate 13‐acetate (>2 nM), over a 16‐hr culture period. Epidermal growth factor at 10 ng/ml induced a small increase in production of IL‐8, but not of IL‐6, whereas bacterial lipopolysaccharide had minimal effects on production of either cytokine. Basal and cytokine‐stimulated IL‐6 and IL‐8 production was inhibited by dexamethasone at concentrations equal to or greater than 1 nM.

CONCLUSION: These findings suggest that amnion may be a significant contributor to the IL‐6 and IL‐8 content of amniotic fluid, and that WISH cells may be a suitable model for the study of cytokine production by amnion epithelial cells.
Amniotic Fluid Granulocyte Colony Stimulating Factor Levels: A Rapid Marker for Diagnosing Chorioamnionitis
Authors: Hoskins Iffath Abbasi , Zandieh Peyman , Schatz Frederick , Lee Christina
Published: September 2011

PROBLEM: To assess the usefulness of amniotic fluid (AF) granulocyte colony stimulating factor (G‐CSF) levels as a rapid marker for diagnosing chorioamnionitis.

METHOD OF STUDY: AF levels were obtained from term and preterm patients with and without chorioamnionitis (CAM). Patients with urinary tract, respiratory tract, and other infections were excluded. Results obtained from the AF G‐CSF assays were compared with those from other parameters used for diagnosing CAM: maternal fever, leukocytosis, tachycardia, fetal tachycardia, AF glucose levels, white blood cell count, Gram stain, and aerobic and anaerobic cultures. The sensitivity, specificity, and predictive values were calculated.

RESULTS: In the uninfected AF samples, G‐CSF levels were present but low, ranging from 400 to 1600 pg/ml. Levels in the infected samples, however, were markedly increased, ranging from 1600 to 14,000 pg/ml; P < 0.05. When a cutoff of 2000 pg/ml was used as a clear marker for CAM, the sensitivity was 67%, the specificity was 100%, and the positive and negative predictive values were 100% and 86%, respectively. The comparison of the other AF G‐CSF laboratory parameters also revealed high sensitivity, specificity, and predictive values for detecting CAM.

CONCLUSION: (i) AF G‐CSF levels are elevated in CAM. (ii) An AF G‐CSF level >2000 pg/ml is a strong positive predictor of CAM. (iii) Elevated AF G‐CSF levels appear to be more reliable in predicting CAM than any other single test currently used in clinical practice.
Expression of Cell Adhesion Molecules in the Extravillous Trophoblast Is Altered in IUGR
Authors: Zygmunt M. , Boving B. , Wienhard J. , Münstedt K. , Braems G. , Bohle R.M. , Lang U.
Published: September 2011

PROBLEM: The invasion of trophoblast cells into the uterine wall and its arterial system is essential for the normal development of pregnancy. Cell adhesion molecules (CAM), such as the immunoglobulin superfamily and integrins, play a crucial role in a number of immunological reactions and in the invasion of the human trophoblast. Intrauterine growth restriction (IUGR) has been associated with abnormal trophoblast invasion. Therefore, the expression of CAM in the extravillous trophoblast of pregnancies complicated by IUGR might be different from normal pregnancies.

METHOD OF STUDY: Normal (n = 21) and IUGR (n = 19) placentas were collected and stored at −70°C. Immunohistochemistry (avidin‐biotin complex peroxidase‐doublestaining) of frozen tissue sections was performed using antibodies specific for the immunoglobulin superfamily vascular adhesion molecule‐1 (VCAM‐1; CD 106), intercellular adhesion molecule (ICAM‐1) (CD 54), ICAM‐2 (CD 102), ICAM‐3 (CD 50), the integrins α2β1, α3β1, α4β1, α5β1, α6β1 and cytokeratin. The percentage of immunopositive extravillous trophoblast cells (EVT) and the intensity of the immunoreactivity for the various CAM and integrin antibodies was assessed.

RESULTS: In IUGR placentas, there was less expression of VCAM‐1 (CD 106), α2β1, α3β1, and α5β1 (P < 0.05) in the extravillous trophoblast than in normal pregnancies. Finally we observed for the first time that ICAM‐3 was expressed on EVT and that its expression was markedly up‐regulated in the EVT of IUGR placentas. No differences were found for ICAM‐1 (CD 54), ICAM‐2 (CD 102), α4β1 and α6β1.

CONCLUSION: Our data show that there are significant differences in the expression of cell adhesion molecules of the extravillous trophoblast from IUGR and normal pregnancies. These differences might reflect changes in the immunological reactions and cell‐cell interactions between mother and the developing fetus which could interfere with fetal growth.
Novel Implications in the Development of Endometriosis: Biphasic Effect of Macrophage Activation on Peritoneal Tissue Expression of Tissue Inhibitor of Metalloproteinase‐1
Authors: Nothnick Warren B. , Soloway Paul D.
Published: September 2011

PROBLEM: Elevated levels of activated macrophages are associated with endometriosis, but their role in the etiology of the disease is uncertain. The current study was undertaken to examine whether activated macrophages could modulate peritoneal tissue expression of the tissue inhibitor of metalloproteinase‐1 (TIMP‐1), which may play a role in the development of endometriosis.

METHOD OF STUDY: Female mice were treated with the macrophage activator lipopolysaccharide (LPS), and peritoneum TIMP‐1 mRNA was examined by Northern blot analysis.

RESULTS: LPS induced a dose‐dependent increase (P < 0.05) in TIMP‐1 mRNA expression at levels of 1 μg (70.0% ± 5.8% greater than the control), 10 μg (83.0% ± 12.0% greater than the control), and 25 μg (100.0% ± 10.0% greater than the control). In contrast, the administration of 50 μg of LPS resulted in a decrease in TIMP‐1 mRNA expression below baseline levels (18.0% ± 6.0% less than the control values).

CONCLUSIONS: Activated macrophages and/or their products modulate peritoneum TIMP‐1 expression. These data suggest that, in addition to their phagocytotic role in the peritoneal cavity, these immune cells also may play a novel role in influencing the ability of the peritoneum to regulate tissue/cell invasion and in the development of endometriosis through TIMP‐1 expression.
Endometrial Leukocytes Are Altered Numerically and Functionally in Women with Implantation Defects
Authors: Stewart‐Akers Ann M. , Krasnow Joel S. , Brekosky Jill , Deloia Julie A.
Published: September 2011

PROBLEM: A significant cohort of women with autoimmune thyroid disease (ATD) also suffer from reduced fertility. The finding that neither exogenous hormones nor donor eggs correct the infertility suggests that the problem involves an inherent endometrial defect.

METHOD OF STUDY: Endometrial leukocyte populations in women with ATD were quantitated by immunohistochemistry. A semi‐quantitative reverse transcriptase polymerase chain reaction (RT‐PCR) technique was used to examine the expression of transforming growth factor (TGF)‐β1, interleukin (IL)‐4, IL‐10, and interferon (IFN)‐γ in the endometrial samples.

RESULTS: A significant increase in the endometrial T‐cell population was observed in women with ATD compared to controls. The relative abundance of IL‐4 and IL‐10 was decreased in women with ATD compared to controls, whereas IFN‐γ was increased. No difference was noted in the abundance of TGF‐β1. The source of cytokine production for IL‐4, IL‐10, and IFNγ was the endometrial leukocytes.

CONCLUSIONS: Both the leukocyte numbers and cytokine expression profile were altered significantly in a well‐defined group of women with implantation defects.
Placental Transport of Maternal Immunoglobulin G
Authors: Palfi Miodrag , Selbing Anders
Published: September 2011

PROBLEM: How does the placental transport of immunoglobulin G (IgG) vary with gestational age?

METHOD OF STUDY: MEDLINE was searched for the appearance of “pregnancy” and “IgG” in the title or abstract from 1966 up to 1995. All publications were reviewed for any kind of IgG data from the fetus and the mother and “known” gestational age. In total, 96 publications from the MEDLINE search fulfilled the search criteria. Seven of ninety‐six publications contained IgG and gestational age data from the fetus and/or the mother. Five other publications that fulfilled the same criteria were also available and were also included. RESULTS: Fetal serum IgG concentrations were found to increase with gestational age according to the formula: fetal IgG = 13.564 − 1.094* gestational age (GA) + 0.026* GA2 (R2 = 0.877). The fetomaternal (f/m) ratio was also found to increase with gestational age according to the formula: f/m IgG = 0.399 − 0.059* GA + 0.003* GA2 − 2.065* 10—5* GA3 (R2 = 0.831).

CONCLUSIONS: The fetal IgG concentrations exceed maternal IgG concentrations after the 35th week of pregnancy. The fetal IgG concentrations are extremely low in the beginning of pregnancy, they exceed 2.5 g/L at >25 pregnancy weeks, and they reach 5 g/L at pregnancy week 30.
Intravenous Immunoglobulin Therapy for Recurrent Spontaneous Abortion: A Meta‐Analysis
Authors: Daya Salim , Gunby Joanne , Clark David A.
Published: September 2011

PROBLEM: Published randomized trials of the use of intravenous immunoglobulins (IVIG) as a treatment for recurrent spontaneous abortion (RSA) have produced conflicting results. The purpose of this study was to conduct a systematic review of the current evidence to evaluate the effectiveness of IVIG for RSA.

METHOD OF STUDY: After a thorough search of the literature, four randomized, doubleblind trials comparing IVIG with placebo for treatment of RSA were included in the metaanalysis. Live birth rates for each treatment group were extracted, and the overall odds ratio (OR) and absolute treatment effect for IVIG were calculated.

RESULTS: Two of the trials showed an increase in successful pregnancy outcome with IVIG treatment and two did not. The overall OR was 1.48 (95% CI, 0.84–2.60) in favor of IVIG, with an absolute treatment effect of 10.1% (95% CI, −4.8–24.6). Excluding pregnancy failures with an obvious cause produced statistically significant results, but this approach may be subject to bias.

CONCLUSION: This meta‐analysis suggests that IVIG may have a role in the treatment of recurrent abortion, but as yet no conclusive evidence is available.
Clinical Significance of Morphologic and Genetic Examination of Spontaneously Aborted Embryos
Authors: Kalousek Dagmar K.
Published: September 2011

PROBLEM: In families with a history of multiple pregnancy losses, the prognosis of future pregnancies is critically dependent on recognizing an accurate pathogenesis of pregnancy loss.

METHOD OF STUDY: Morphological and genetic evaluation of products of conception provides necessary information for clinicians. Modern molecular cytogenetic techniques such as in situ hybridization and comparative genomic hybridization allow cytogenetic diagnosis even when aborted tissues are nonviable.

RESULTS: Correlation of morphological and cytogenetic findings allows distinction between developmental defects associated with chromosomal syndromes and other pathogenesis such as environmental teratogens, and Mendalian or multifactorial inheritance.

CONCLUSION: Pathologists have the responsibility of ensuring that the answers to questions, such as why the failure occurred or whether there is any increased chance of having an abnormal liveborn infant in a future pregnancy, are obtained and that the information is communicated to the parent's obstetrician, geneticist, and family physician.
High Levels of Anticardiolipin Antibodies in Patients with Abnormal Embryo Morphology Who Attended an In Vitro Fertilization Program
Authors: Azem Foad , Geva Eli , Amit Ami , Lerner‐Geva Liat , Shwartz Tamar , Ben‐Yosef Dalit , Yovel Israel , Lessing Joseph B.
Published: September 2011

PROBLEM: Recently, it has been suggested that anticardiolipin antibodies (ACAs) may serve as possible markers for reproductive failure. The association between ACAs and embryo morphology in patients undergoing in vitro fertilization (IVF) was investigated.

METHOD OF STUDY: This prospective study comprised 117 patients with either tubal factor or unexplained infertility. Embryo morphology was blindly scored from I to IV according to blastomere regularity and the presence of fragments. Anticardiolipin antibodies (immunoglobulin [Ig] G and IgM) were detected.

RESULTS: Anticardiolipin antibodies were found in 26 (50%) of the 52 patients with abnormal morphology, compared with 13 (20%) of the 65 patients with normal embryo morphology (P = 0.001). No statistically significant differences were found between the prevalence of ACAs among patients with tubal factor and those with unexplained infertility (29.6% and 36.5%, respectively).

CONCLUSIONS: Our study shows an association between embryo morphology and the presence of ACAs. This association may explain the low implantation rate and early pregnancy loss in patients with ACAs.
Female Reproductive Tract Immunity in Bovine Trichomoniasis
Authors: Corbeil Lynette B. , Anderson Mark L. , Corbeil Robert R. , Eddow John M. , BonDurant Robert H.
Published: September 2011

PROBLEM: Mechanisms of protective immunity in the female reproductive tract are poorly understood. For sexually transmitted diseases, bovine trichomoniasis is a useful model because il resembles human trichomoniasis to some extent, and antibodies play an important role in protection against these extracellular parasites. Protective efficacy was compared in animals with genital responses of predominantly immunoglobulin G (IgG) or predominantly IgA antibodies to a purified surface antigen ofTritrichomonas foetus.

METHOD OF STUDY: Immunization of mice by various routes with immunoaffinity‐purifiedT. foetussurface antigen (TF1.17) or killed cells was used to define the best routes and antigen combinations to give predominantly IgG or IgA antibodies to TF1.17 antigens in genital secretions. Cattle were then immunized either subcutaneously (SC) two times with TF1.17 antigen and once SC with killedT. foetusor twice SC with TF1.17 antigen and once intravaginally with killedT. foetus.All immunizations were in Quil A adjuvant. Controls were not immunized. Animals were challenged intravaginally with 106T. foetus3 weeks after the third immunization. Vaginal mucus was collected weekly for culture and antibody assays. Serum was collected weekly, and uterine secretions were collected at 10 weeks post‐challenge. Tissues were fixed at 10 weeks also.

RESULTS: Murine studies showed systemic priming with vaginal boosting gave the highest genital IgA responses. In cattle, systemic immunization (group S) induced high IgG1 antibody levels in vaginal secretions. Systemic priming with vaginal boosting (group S/V) primed for an anamnestic vaginal IgA response after challenge withT. foetus.Cattle with predominantly IgG or predominantly IgA responses in vaginal secretions either did not become infected or cleared infection faster than controls. Uterine IgA responses at 10 weeks were highest in the vaginally boosted group, but other responses were not different from the controls at this time point. Microscopic examination of genital tissues showed subepithelial infiltration of mononuclear cells in all groups. Lymphoid aggregates or nodules were detected in vaginal sections in cattle of groups S/V and C as well as in uterine sections of all animals in all three groups.

CONCLUSIONS: Both IgG and IgA antibodies toT. foetussuperficial antigen were associated with protection. The timing of the response was related to the time of clearance. Lymphoid organization in the vagina and uterine tissues suggested development of mucosal inductive sites.
Influence of the Estrous Cycle on the Presence and Distribution of Immune Cells in the Rat Reproductive Tract
Authors: Kaushic Charu , Frauendorf Elaine , Rossoll Richard M. , Richardson Jan M. , Wira Charles R.
Published: September 2011

PROBLEM: Previous studies have shown that the uterus and vagina contain cells that can present antigen to ovalbumin‐specific T‐cells. The objective of the present study was to systematically characterize the immune cells [major histocompatibility complex (MHC) class‐II+, macrophages, granulocytes, dendritic cells, and CD8+ cells] present in the uterus and vagina of the rat and to examine their distribution at various stages of the estrous cycle.

METHOD OF STUDY: Uterine and vaginal tissues from female rats were selected at various stages of the estrous cycle and were examined by immunohistochemical analysis. MHC class‐II (Ia)‐positive cells were detected using the OX‐6 monoclonal antibody; macrophages, granulocytes, and dendritic cells were detected by OX‐41 monoclonal antibody and CD8‐positive T‐cells were identified by OX‐8 monoclonal antibody.

RESULTS: Immunohistochemical analysis showed cycle‐dependent changes in the immune cell populations in the uterus and vagina. Ia+ cells, macrophages, granulocytes, and dendritic cells were present in large numbers in the stroma of the endometrium and around the glandular epithelium in the uterus at estrus, the stage of the reproductive cycle when estradiol levels are known to be high, relative to those seen at diestras, when estrogen levels are low and progesterone is the predominant hormone. CD8+ cells were observed in the uterus interspersed between glandular epithelial cells at estrus. Immune cells were more numerous in the vagina, relative to the uterus. OX‐6‐ and OX‐41‐positive cells were present in greater numbers in the subepithelial layers of the vagina at diestras, in contrast to estrus.

CONCLUSION: This study demonstrates that a variety of immune cells are present in the reproductive tract and that their number and distribution vary in a tissue‐specific manner with the stage of the estrous cycle.
Evidence That Folliculo‐Stellate Cells Mediate the Inhibitory Effect of Japanese Kampo Medicine, Unkei‐To, on Growth Hormone Secretion in Rat Anterior Pituitary Cell Cultures
Authors: Koike K. , Zhang Z. X. , Sakamoto Y. , Miyake A. , Inoue M.
Published: September 2011

PROBLEM: Cytokine‐induced neutrophil chemoattractant (CINC) was reported to influence anterior pituitary hormone release. We recently found that Unkei‐to, one of the Japanese Kampo medicines, stimulated CINC secretion by rat anterior pituitary cells and the pituitary folliculo‐stellate (FS)‐like cell line (TtT/GF). Therefore, the effect of Unkei‐to on growth hormone (GH) secretion by rat anterior pituitary cells was investigated.

METHOD OF STUDY: Dispersed normal anterior pituitary cells, the folliculo‐stellate‐like cell line TtT/GF, and the GH3 cell were used to test the effect of Unkei‐to on GH secretion. In reconstitutive coculture experiments, TtT/GF cells were mixed with GH3 cells at a ratio (TtT/GF cells: GH3 cells) of 1:99. From this mixture, cells were seeded onto plates at a density of 104 cells/well and were cultured for 5 days. The cells were then used in the experiments.

RESULTS: Unkei‐to at 20 μg/ml significantly inhibited GH secretion by normal anterior ptuitary cells within 12 hr of incubation. In contrast Unkei‐to stimulated GH secretion by GH3 cells in a time‐ and dose‐dependent manner, suggesting that an accessory cell type was involved. To assess the contribution of CINC as a paracrine factor, an experiment using a reconstitutive coculture system was performed, and Unkei‐to was found to inhibit GH secretion when GH3 cells were cocultured with TtT/GF cells. The addition of anti‐CINC antibody to the reconstitutive coculture system antagonized Unkei‐to‐inhibited GH secretion.

CONCLUSION: CINC, which was secreted from FS cells by Unkei‐to, may be responsible for mediating the inhibitory effect of Unkei‐to on GH secretion by rat anterior pituitary cells.
The Use of Combined Heparin/Aspirin and Immunoglobulin G Therapy in the Treatment of In Vitro Fertilization Patients With Antithyroid Antibodies
Authors: Sher Geoffrey , Maassarani Ghanima , Zouves Christo , Feinman Michael , Sohn Sae , Matzner William , Chong Penny , Ching Wendell
Published: September 2011

PROBLEM: To compare the effect of heparin/aspirin therapy alone vs. heparin/aspirin in combination with intravenous immuno‐globulin (IVIg) immunotherapy on in vitro fertilization (IVF) outcome of patients who test positive for antithyroid antibodies (ATAs).

METHOD OF STUDY: Eighty‐two women younger than 40 years of age whose infertility was related exclusively to female causes were evaluated. All tested positive for organ‐specific antithyroid antibodies (antimicrosomal and/or antithyroglobulin antibodies), but negative for antiphospholipid antibodies. Thirty‐seven of these women (group A) received H/A alone, whereas 45 (group B) received heparin/aspirin in combination with IVIg.

RESULTS: Ten (27%) of women in group A and 23 (51%) of women in group B achieved live births after completion of a single IVF/embryo transfer cycle (P = 0.027).

CONCLUSION: We conclude that IVIg therapy significantly improves IVF success rates in ATA+ women.
Adenosine Deaminase and Human Reproduction: A Comparative Study of Fertile Women and Women with Recurrent Spontaneous Abortion
Authors: Nicotra M. , Bottini N. , Grasso M. , Gimelfarb A. , Lucarini N. , Cosmi E. , Bottini E.
Published: September 2011

PROBLEM: We have investigated the possible role of adenosine deaminase (ADA) genetic polymorphism in human fertility through a comparative study of couples with recurrent spontaneous abortion (RSA) and healthy puerperae.

METHOD OF STUDY: Adenosine deaminase phenotype has been determined in 209 women with repeated episodes of unexplained spontaneous abortion (RSA) and their husbands, as well as in 115 healthy pregnant women from the population of Rome. An independent sample of 286 puerperae along with their newborn infants in the population of Penne was also studied.

RESULTS: The proportion of carriers of ADA*2 allele, which is associated with the lowest enzymatic activity, is lower among women with RSA than among healthy pregnant women from the same population of Rome. Preliminary observations suggest a protective effect of ADA*2 against the development of autoantibodies in RSA. Such an effect seems to be mediated by an interaction with ABO blood groups. In the population of Penne the proportion of women carrying ADA*2 allele is higher among those who have had two or more previously born children than among women with only one or no children.

CONCLUSIONS: The data suggest that women carrying the ADA*2 allele are better protected against the spontaneous loss of embryos and have a higher fertility rate.
How Might Pregnancy Immunize Against Breast Cancer?
Authors: Botelho Fernando , Clark David A.
Published: September 2011

PROBLEM: This study investigated how pregnancy might protect against breast cancer. METHOD OF STUDY: A critical review of the literature was done.

RESULTS/CONCLUSIONS: Support for an active role in pregnancy immunizing against breast cancer comes from case studies demonstrating a reciprocal correlation between pregnancy and breast cancer as well as recent experiments supporting the fetal antigen hypothesis that confirms the presence of a tumor‐specific antigen, MUC1, on both fetal and breast cancer tissues. Multiparous women also generate anti‐MUC1 major histocompatibility complex‐restricted cytotoxic T cell cytolytic activity against MUC1‐bearing tumor cell lines. Careful investigation of the fetal antigen hypothesis and tolerogenic mechanisms may lead to effective vaccination protocols against breast cancer and other cancers.
Regain of Fertility and Normality of Progeny Born During Below Protective Threshold Antibody Titers in Women Immunized With the HSD‐hCG Vaccine
Authors: Singh Meharban , Das S.K. , Suri S. , Singh Om , Talwar G.P.
Published: September 2011

PROBLEM: Phase II clinical trials with the heterospecies dimer of βhCG and α‐subunit of ovine luteinizing hormone (HSD)‐human chorionic gonadotropin (hCG) vaccine showed that pregnancy was prevented at and above 50 ng/ml titers, whereas conceptions occurred below 35 ng/ml of hCG bioneutralization capacity. The effect of below‐protective threshold anti‐hCG antibodies on the progression of pregnancy and the normality of progeny was studied.

METHOD OF STUDY: Four women enrolled by informed consent in the Phase II trials. The women studied were immunized with the HCG vaccine and did not receive booster injections in consideration of their desire to have another child. They were examined clinically at least once every month until delivery. The babies were followed up from 2 to 3.5 years, and their anthropometric indices were compared with elder siblings.

RESULTS: The pregnancies progressed to 35 to 38 weeks with the usual obstetric problems, and the children developed normally.

CONCLUSIONS: The anti‐fertility effect of the HSD‐hCG vaccine is reversible and low titers of antibodies below the protective threshold have no apparent side effects on the progression of pregnancy and on the early development of the progeny.
Suppression of Natural Killer Cell Activity and Interleukin‐2 Concentration of Serum Obtained from In Vitro Fertilization‐Embryo Transfer Patients
Authors: NAKAMURA YOSHIHIRO , FUJINO YUJI , OZAKI KOUJI , UEDA KAYOKO , OZAKI AYAKO , ITOH FUMITOSHI , OGITA SACHIO
Published: September 2011

PROBLEM: The effect of serum obtained from in vitro fertilization‐embryo transfer (IVF‐ET) patients on healthy volunteers' natural killer (NK) cell activity was evaluated. We also measured interleukin (IL)‐2 concentration with IVF‐ET patients' serum and clarified the relationship between IL‐2 levels and the suppressive effect on NK cell activity.

METHOD OF STUDY: A retrospective nonrandomized clinical study was performed. The suppressive effect on NK cell activity and IL‐2 concentrations was measured with serum obtained from 30 pregnant and 30 nonpregnant women during an IVF‐ET procedure. The suppressive effect of the serum on NK cell activity was evaluated by the formula that we defined in our previous study.

RESULTS: The suppression of NK cell activity was significantly higher in the nonpregnant women than in the pregnant women (P < 0.05); however, IL‐2 concentration did not differ. There was a positive correlation between the suppression of NK cell activity and IL‐2 levels in the pregnant women, but no significant correlation in the nonpregnant women.

CONCLUSIONS: These results suggest that the suppression of NK cell activity may be one of the prognostic factors for IVF‐ET. In addition, we speculate that an unidentified humoral factor other than IL‐2, which could increase NK cell activity, might exist in the serum of the nonpregnant patient.
Expression of Vimentin and Cytokeratin in Eutopic and Ectopic Endometrium of Women with Adenomyosis and Ovarian Endometrioma
Authors: SONG IN OK , HONG SUNG RAN , HUH YOUNGBUHM , YOO KEUN JAI , KOONG MI KYOUNG , JUN JONG YOUNG , KANG INN SOO
Published: September 2011

PROBLEM: To determine the expression of vimentin and cytokeratin in eutopic and ectopic endometrium of women with both adenomyosis and ovarian endometrioma and to evaluate their cyclic changes during the menstrual cycle.

METHOD OF STUDY: Twenty patients requiring hysterectomy with salpingo‐oophorectomy were studied by immunohistochemistry according to their menstrual cycles.

RESULTS: Cyclic expression of vimentin was noted in eutopic endometrium and adenomyosis, but not in endometrioma. Cytokeratin expression did not change during the menstrual cycles. The mean intensities of epithelial vimentin were significantly different from each other, being the lowest in endometrioma, intermediate in adenomyosis, and the highest in eutopic endometrium. There was no significant difference in intensities of cytokeratin between adenomyosis and endometrioma, but these intensities were significantly lower than that of eutopic endometrium.

CONCLUSIONS: Lower intensities of cytokeratin in adenomyosis and endometrioma than in eutopic endometrium suggest that the ectopic endometria may have a lower degree of differentiation regardless of the site. The lower intensity of epithelial vimentin in endometrioma than in adenomyosis during the proliferative phase may reflect decreased functional activity, probably because of a pressure effect on the lining epithelium within the endometrioma.
Do Autoantibodies to Sperm Reduce Fecundity? A Mini‐Review in Historical Perspective
Authors: HJORT TAGE
Published: September 2011

PROBLEM: The paradox that early studies of antisperm antibodies in men showed a strong correlation between titers of circulating antibodies (essentially immunoglobulin [Ig] G) and reduction in conception rates, whereas more recent studies have indicated that the antifertility effect is mainly (or exclusively?) associated with IgA antibodies, impairing sperm migration through cervical mucus, was studied.

METHOD OF STUDY: Relevant literature focusing on antibodies on ejaculated sperm was analyzed.

RESULTS: Direct mixed antiglobulin reaction (MAR) and immunobead‐binding tests are excellent and sensitive techniques for demonstrating antibodies of the IgG and IgA classes on sperm, and they have revealed that IgA antibodies are, with very rare exceptions, found only when IgG antibodies are also present. However, these tests tell little about the amounts of antibodies present, and attempts to measure quantitatively the amounts of Ig on sperm have indicated higher levels of IgA than IgG (despite the strongest MAR reactivities for IgG).

CONCLUSIONS: The patients with high levels of IgA to their sperm are mainly men with strong immune responses and, therefore, also high antibody titers in serum. Apparently, the locally produced IgA antibodies reach the sperm and occupy the binding sites before the main bulk of IgG reaches the seminal compartment with the prostatic fluid.
Altered Modulation of the In Vitro Antibody Synthesis by Placental Factors from the CBA/J × DBA/2 Abortion‐Prone Mating Combination
Authors: Miranda S. , Borel I. Malán , Margni R.
Published: September 2011

PROBLEM: The in vitro immunomodulating effect of placental culture supernatants (PSs) obtained from two H‐2k × H‐2d allogeneic crossbreedings, the CBA/J × DBA/2 abortion‐prone mating combination, and the reproductively normal pregnancy CBA/J × BALB/c crossbreeding were compared, and the influence of previous deliveries was evaluated. The behavior of placentae obtained from CBA/J females with two previous pregnancies by BALB/c males was also investigated.

METHOD OF STUDY: Supernatants of cultures of murine placentae were added to a mouse immunoglobulin (Ig) G1 hybridoma culture which produced anti‐dinitrophenol (anti‐DNP) antibodies. The quantity of monoclonal antibody produced, the nature of these antibodies, and the proliferation of the hybridoma cells were studied.

RESULTS: CBA/J × DBA/2 placental factors obtained from multiparous females induced a diminished asymmetric IgG antibody production without varying the quantity of antibody produced. In contrast, PSs obtained from the nonresorption‐prone CBA/J × BALB/c mating combination with the same number of previous deliveries enhanced the production of both symmetric and asymmetric anti‐DNP molecules and also increased the proportion of asymmetric blocking monoclonal antibodies (mAbs) synthesized by the hybridoma. Both of the PSs analyzed had induced similar inhibition of 3H‐thymidine uptake. PSs obtained from the abortion‐prone mating combination whose CBA/J females had two previous pregnancies by BALB/c males showed similar immunomodulating effects to those observed using multiparous CBA/J × BALB/c placentae.

CONCLUSIONS: We propose that the placenta produces soluble factors that participate in the regulation of antibody synthesis by the mother during gestation. Such a placental immunomodulating effect appears to be altered in the CBA/J × DBA/2 abortion‐prone mating combination and could be corrected by previous pregnancies by BALB/c males. These observations suggest that placental factors would be relevant to the protection of the fetus and might play an important role in the immune equilibrium between mother and fetus. Asymmetric antibody production as a Th2 responsiveness was also discussed.
Mechanisms of Action of Major‐Histocompatibility‐Complex — Linked Genes Affecting Reproduction
Authors: Gill Thomas J.
Published: September 2011

PROBLEM: To provide insight into the mechanisms of action of the major‐histocompatibility‐complex (MHC)‐linked genes affecting reproduction.

METHOD OF STUDY: The data were obtained using a variety of cellular and molecular techniques in experimental animals and from population genetic studies in humans.

RESULTS: In the mouse, the preimplantation embryonic development (Ped) locus, whose functional gene is Q9, regulates fast and slow cleavage of the early embryo. There is also evidence for a growth and reproduction complex (Grc)‐like region from serologic, molecular, and cytogenetic studies. In the human, the human leukocyte antigen (HLA)‐G gene has been associated with an increased rate of embryonic cleavage in those embryos that express the HLA‐G antigen. Sharing of HLA antigens in couples has been associated with recurrent spontaneous abortions, gestational trophoblastic tumors, and unexplained infertility. Detailed mapping studies showed that the genes responsible are not the HLA genes themselves, but genes closely linked to the HLA‐DR‐DQ‐B genes. The HLA region genes can interact epistatically with the C3 allele of transferrin to increase the incidence of fetal loss. In the rat, the Grc region, which is closely linked to the MHC, has been associated with embryonic loss, growth defects, and susceptibility to chemical carcinogens. The Grc can interact epistatically with the tail anomaly lethal (Tal) gene or the hood restriction (Hre) gene to enhance these effects.

CONCLUSIONS: There are two basic mechanisms for the effects of MHC‐linked genes on reproduction and development: individual gene effects (Ped [Q9], HLA‐G) and extended genetic effects (MHC‐linked genes in the rat [Grc] and in the human). The nature of these genetic effects, particularly the MHC‐linked effects, can also provide some insight into the different theories of human origins: These effects are most consistent with the monogenic theory.
Natural Killer (NK) Cell Subsets and NK Cell Cytotoxicity in Women with Histories of Recurrent Spontaneous Abortions
Authors: Gilman‐Sachs Alice , DuChateau Brian K. , Aslakson Cheryl J. , Wohlgemuth Ghislaine P. , Kwak Joanne Y. , Beer Alan E. , Beaman Kenneth D.
Published: September 2011

PROBLEM: Natural Killer (NK) cell measurement and NK cytotoxicity are two measurements for assessing the cellular immune response. Both of the techniques have been reported to be prognostic for women with recurrent spontaneous abortion (RSA), We evaluated the two methods to determine the relationship of the two assays. Because both methods portend to evaluate the same process, the previous clinical data suggested that the methods evaluate the same phenomena. We undertook these studies to determine whether simple NK cell counts may be sufficient in the evaluation of NK activity in RSA.

METHOD OF STUDY: The NK cell cytotoxicity at effector‐to‐target ratios of 50:1 and 25:1 was determined using a flow cytometric NK cell cytotoxicity assay. These values were then correlated with the percentages and absolute counts of three peripheral blood NK cell subsets.

RESULTS: The data indicate that the flow cytometric assay is reproducible and precise and can be successfully used to evaluate patient samples. Linear regression analysis indicated a lack of correlation between peripheral blood NK cell cytotoxicity and percentages or absolute counts of ***CD56+CD16+, CD56+CD16 — or CD3+CD56+ lymphocyte subsets (range of correlation coefficients, 0.1–0.3).

CONCLUSIONS: NK cell cytotoxicity and peripheral blood NK cell values measure different aspects of NK cells and do not correlate. These data indicate that simple enumeration of NK cells may not be sufficient in the evaluation of NK cells in RSA.
Increase in the Production of Interleukin‐6, Interleukin‐10, and Interleukin‐12 by Lipopolysaccharide‐Stimulated Peritoneal Macrophages from Women with Endometriosis
Authors: Wu Ming‐Yih , Ho Hong‐Nerng , Chen Shee‐Uan , Chao Kuang‐Han , Chen Chin‐Der , Yang Yu‐Shih
Published: September 2011

PROBLEM: To verify whether the peritonea] macrophage (PM) is activated in endometriosis.

METHOD OF STUDY: We examined the synthesis of nitric oxide (NO), total antioxidant, interleukin (IL)‐6, IL‐10, and IL‐12 by cultured PMs, which were either unstimulated or stimulated with lipopolysaccharide (LPS), from women with endometriosis (early, n ? 12; advanced, n ? 11) or without endometriosis (n = 13). RESULTS: After stimulation with 2 ng/mL LPS for 24 hr, PMs from women with early‐stage endometriosis secreted more NO, IL‐6, and IL‐10 than the controls. Higher IL‐12 levels were noted in women with advanced endometriosis when compared with the controls. After 2 ng/mL‐LPS stimulation for 24 hr, we also detected higher total antioxidant levels in the advanced‐endometriosis group than those in the early‐endometriosis group.

CONCLUSION: The increased production of IL‐6, IL‐10, and IL‐12 by stimulated PMs confirmed previous observations that the PM is the principle source of these cytokines in peritoneal fluid.
Interleukin‐6 Levels in Co‐culture of Human In Vitro Fertilization Embryos with Vero Cells Are Not Predictive of Future Successful Development
Authors: Laprée‐Delage Geneviève , Volante Monique , Frydman René , Chaouat Gérard
Published: September 2011

PROBLEM: In an attempt to predict successful embryo transfer and implantation, we measured interleukin (IL)‐6 levels in culture supernatants of co‐cultured preimplantation human embryos. We tested whether all in vitro—fertilized human embryos in co‐cultures do secrete IL‐6, and whether there was any difference in such production between embryos that successfully reached the blastocyst stage and blocked embryos. We also addressed the question of IL‐6 secretion by co‐culture support cells, namely Vero cells themselves.

METHOD OF STUDY: Each fertilized oocyte was cultured individually and transferred in culture wells supplemented with a feeder layer of Vero cells at day 2. In vitro IL‐6 production was measured by bioassay of the culture media.

RESULTS: Because Vero cells themselves secrete IL‐6, it became impossible, in co‐culture, to quantify production of IL‐6 by the sole embryos. On the other hand, the co‐culture technique has shown us that embryos are likely to consume IL‐6. There was no difference between blastocysts and blocked embryos.

CONCLUSIONS: IL‐6 levels in human embryo co‐cultures do not correlate with future successful embryo transfer.
Possible Susceptibility of the HLA‐DPB1*0402 and HLA‐DPB1*04 Alleles to Unexplained Recurrent Abortion: Analysis by means of Polymerase Chain Reaction–Restricted Fragment Length Polymorphism Method
Authors: Takakuwa Koichi , Hataya Isao , Arakawa Masato , Kikuchi Akira , Higashino Masahiko , Yasuda Masako , Kurabayashi Takumi , Tanaka Kenichi
Published: September 2011

Takakuwa K, Hataya I, Arakawa M, Kikuchi A, Higashino M, Yasuda M, Kurabayashi T, Tanaka K. Possible susceptibility of the HLA‐DPB1*0402 and HLA‐DPB1*04 alleles to unexplained recurrent abortion – analysis by means of polymerase chain reaction–restricted fragment length polymorphism method. AJRI 1999; 42:233–239 © Munksgaard, Copenhagen

PROBLEM: To clarify whether HLA‐DP antigens are associated with patient population of unexplained recurrent abortion.

METHOD OF STUDY: The frequency of HLA‐DPB1 alleles in patients with unexplained recurrent abortion, and the compatibility of HLA‐DPB1 alleles between patient couples, were studied using a polymerase chain reaction (PCR)–restricted fragment length polymorphism (RFLP) method. Thirty patients who had a history of unexplained primary recurrent abortion, and their husbands, were typed for HLA‐DPB1 genotype. Two hundred and ninety‐nine base pair fragments from the second exon of HLA‐DPB1 genes were selectively amplified using the PCR‐primers. After amplification, the DNAs were digested with restriction endonucleases, and subjected to electrophoresis in a 12% polyacrilamide gel to determine HLA‐DPB1 genotype.

RESULTS: The frequency of HLA‐DPB1*0402 and DPB1*04 alleles in the patient group (n = 30) was significantly increased, as compared to that in the normal fertile women (n = 30). The frequency of HLA‐DPB1*04 allele in the patient group was significantly increased, as compared to that in the general population (n = 112). No significant compatibility of HLA‐DPB1 alleles could be observed between patient couples and normal fertile couples.

CONCLUSION: These findings suggest a possible new class II association with patient population of unexplained recurrent abortion.
Isolation and Biological Characterization of Boar Sperm Capacitation‐Related Antigen
Authors: Mollova Margarita , Djarkova Tsvetelina , Ivanova Maria , Stamenova Maria , Kyurkchiev Stanimir
Published: September 2011

Mollova M, Djarkova T, Ivanova M, Stamenova M, Kyurkchiev S. Isolation and biological characterization of boar sperm capacitation‐related antigen. AJRI 1999; 42:254–262 © Munksgaard, Copenhagen

PROBLEM: Monoclonal anti‐capacitated sperm antibody has been used as a probe to identify, isolate, and characterize specific, fertilization‐related antigen with some characteristic features that point to its possible significance in immunocontraception.

METHOD OF STUDY: Fast protein liquid chromatography (FPLC), enzyme‐linked immunosorbent assay (ELISA), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE), and isoelectrofocusing were used for isolation, immunochemical and physicochemical characterization of a monoclonal antibody (mAb) 1F10 cognate antigen. Sperm‐zona pellucida binding and hemizona assay were used for testing the biological roles of mAb 1F10 and Ag 1F10 in boar and human fertilization processes.

RESULTS: The Ag 1F10 was found to be eluted in the eighth protein peak of FPLC‐fractionated Nonidet P40 (NP40) extracts of capacitated boar spermatozoa. The SDS‐PAGE and immunoelectrofocusing experiments showed that Ag 1F10 is a protein composed of a single peptide chain with a relative molecular mass of 68/70 kDa and an isoelectric point of 3.5. It was demonstrated that the zona binding activity of spermatozoa preincubated in the presence of mAb 1F10 was significantly inhibited both in porcine and human in vitro fertilization (IVF) systems. A dose‐dependent manner of inhibition of the sperm/ligand activities of porcine and human zona pellucida was observed when the effect of purified Ag 1F10 was investigated by its preincubation with zona pellucida.

CONCLUSIONS: It is assumed that the protein bearing the epitope recognized by mAb 1F10 may be accepted as one of the molecules with receptor function in sperm‐zona pellucida interaction during fertilization.
Circulating Interferon‐γ‐ and Interleukin‐4‐Secreting Cells in Recurrent Spontaneous Abortions
Authors: Palfi Miodrag , Jablonowska Barbara , Matthiesen Leif , Ernerudh Jan
Published: September 2011

PROBLEM: Are recurrent spontaneous abortions (RSAs) associated with deviation of circulating cytokine‐secreting cells?

METHOD OF STUDY: Interferon (IFN)‐γ‐ and interleukin (IL)‐4‐secreting cells were identified by enzyme‐linked immunospot (ELISPOT) in blood from 34 women with RSA. Samples were taken before pregnancy and/or pregnancy weeks 7–10, 17–20, and after terminated pregnancy. Eleven healthy primigravidae and 10 nonpregnant women served as controls.

RESULTS: No significant difference in numbers of IFN‐γ‐ and IL‐4‐secreting cells was noted within the RSA group when abortions and successful pregnancies were compared in samples taken before pregnancy. The number of IFN‐γ‐ as well as IL‐4‐secreting cells in pregnancy weeks 17–20 in the RSA group was significantly higher compared with before pregnancy, pregnancy weeks 7–10, and after pregnancy. In samples from non‐pregnant women, the number of IFN‐γ‐ and IL‐4‐secreting cells was significantly higher in the RSA group compared with controls.

CONCLUSIONS: Our results do not indicate a systemic shift in the general balance between T helper 1‐ and T helper 2‐type cytokine pattern. A local shift at the fetomaternal interface seems more probable.
Oral Vaccination of White‐Tailed Deer Using a Recombinant Bacillus Calmette‐Guérin Vaccine Expressing the Borrelia burgdorferi Outer Surface Protein A: Prospects for Immunocontraception
Authors: Miller Lowell A. , Johns Brad E. , Elias Donald J. , Killian Gary J.
Published: September 2011

PROBLEM: Reduction of excess numbers of white‐tailed deer (Odocoileus virginianus) is a prime example of a potential use for immunocontraception as a means of wildlife population management. Oral vaccination appears to be the most pragmatic way to deliver immunocontraceptive vaccines to free‐roaming populations of deer, but there was little, if any, prior evidence that oral vaccination is a viable concept in deer.

METHOD OF STUDY: We used live Bacillus Calmette‐Guérin (BCG) in a recombinant form (rBCG), which expressed Borrelia burgdorferi outer surface protein A, to test whether deer vaccinated orally with a specific antigen expressed in a live vector produce detectable antibody titers.

RESULTS: The data indicate that oral vaccination of deer with an expressed antigen is feasible, as demonstrated by peak antibody titers to the expressed antigen. Also, peak titers measured by enzyme‐linked immunosorbent assay were highest in orally vaccinated deer: 1600 in deer vaccinated by injection and 6400 in those vaccinated orally.

CONCLUSIONS: The results of this study demonstrate that it is feasible to vaccinate deer orally with a live vector.
Review: Vertical Transmission of HIV: Parameters Which Might Affect Infection of Placental Trophoblasts by HIV‐1: A Review
Authors: Moussa Marléne , Mognetti Barbara , Dubanchet Sylvie , Menu Elisabeth , Roques Pierre , Gras Gabriel , Dormont Dominique , Barre‐Sinoussi Françoise , Chaouat Gérard
Published: September 2011

PROBLEM: To understand the mechanisms preventing and/or facilitating maternofetal transmission of human immunodeficiency virus (HIV)‐1 across the placenta during pregnancy.

METHODS OF STUDY: Current experimental data were reviewed.

RESULTS AND CONCLUSIONS: The data about the production of cytokines by placental cells and expiants, taken together with information indicating selective passage of certain HIV‐1 variants across the placental trophoblast, suggest an intricate regulatory network operating at the fetomaternal interface. The data show a differential differentiation of early and late trophoblasts, as far as HIV entry routes are concerned. We believe this explains the relative predominance of the early infection window, as far as in utero infection is concerned. Whether such a differentiation state can be transiently induced on term placental trophoblasts by several differentiation agents, including cytokines, is being investigated. Whatever the results may be, it is obvious that infection of placental cells is an excellent model of passage infection by HIV of through a mucosal barrier.