bluuebunny/crossref_metadata_2025 huggingface dataset:
Distinct Expression of Cytokines and Mitogenic Inhibitory Factors in Semen of Fertile and Infertile Men
Authors: HULEIHEL MAHMOUD , LEVY AYELET , LUNENFELD EITAN , POTASHNIK GAD , GLEZERMAN MAREK , HOROWITZ SHULA
Published: September 2011

PROBLEM: To assess the effect of seminal plasma (SP) of fertile and infertile men on leukocyte mitogenic response, and the capability of sperm cells to produce IL‐1.

METHODS: This study included four groups: fertile men (donors, normal), infertile men with azoospermia (azoo), oligo‐terato‐asthenozoospermia (OTA), and OTA with genital infection (OTA‐inf). Mouse spleen cell proliferation in response to lipopolysaccharide (LPS) or Concanavalin‐A (Con‐A) was examined in the presence of SP from the above four groups. Supernatants (sup) and lysates (lys) of sperm cells from fertile and oligoteratoasthenospermic (OTA) men were evaluated for IL‐1 bioactivity by specific bioassay.

RESULTS: Seminal plasma (SP) of the four groups were shown to inhibit the mitogenic response of mouse spleen cells to LPS and Con‐A. SP of fertile men was significantly more inhibitory than SP from infertile men. Sperm cells from fertile and OTA infertile men constitutively produced IL‐1. Sperm cells of both groups produced similar levels of IL‐1 as examined in the supernatants and lysates.

CONCLUSIONS: Seminal plasma of fertile men had more inhibitory mitogenic activity than that of OTA. Sperm cells constitutively produce IL‐1. It is possible that the factors involved in this inhibition are not only anti‐proliferative immune factors. Cytokines and inhibitory factors of mitogenesis in the seminal plasma may be involved in the physiology and pathophysiology of sperm functions and thus affect male fertility.
Correlation of Basal Menses CA‐125 Levels and 6 Month Pregnancy Rates in Women Undergoing Treatments for Infertility Without Assisted Reproductive Methods
Authors: CHECK JEROME H. , COHEN RACHEL , PEYMER MARK , RESNICK MICHAEL , SURYANARAYAN CHITOOR
Published: September 2011

PROBLEM: The objective of this study was to evaluate the correlation of menstrual CA‐125 levels with pregnancy rates (PRs) after 6 months of treatment for infertility.

METHOD: The sample consisted of a heterogenous group of 160 women who sought treatment for infertility. Treatments include progesterone supplementation, donor insemination, intrauterine insemination, and ovulation induction therapy. No laparoscopies were done during the study period. A baseline CA‐125 level was drawn during menses before the initiation of therapy. Patients were followed for 6 months of treatment or until a pregnancy was achieved.

RESULTS: There was no difference in the 6 month PR or viable PR by CA‐125 level.

CONCLUSIONS: Elevated CA‐125 levels are not predictive of poor fertility potential at least during the first 6 months of infertility therapy. Even though these higher levels sometimes suggest that endometriosis is present, the data suggest that correction of male factor, cervical factor or ovulation factor provides effective PRs without the need for laparoscopic intervention.
Expression of an Immunomodulatory Protein Known as Progesterone Induced Blocking Factor (PIBF) Does Not Correlate With First Trimester Spontaneous Abortions in Progesterone Supplemented Women
Authors: Check Jerome H. , Ostrzenski Adam , Klimek Rudolf
Published: September 2011

PROBLEM: An immunomodulatory protein known as the progesterone induced blocking factor (PIBF) has been found to positively correlate with early pregnancy beta human chorionic gonadotropin (B‐hCG) levels. The study presented herein evaluated PIBF levels from conception to the end of the first trimester to determine if lower levels will correlate with first trimester spontaneous abortions (SAB).

METHOD: Progesterone induced blocking factor expression by lymphocytes measured using an immunocytochemistry method was compared in pregnant women with ongoing vs. failed pregnancies.

RESULTS: There were no differences in the proportion of women having lymphocytes expressing PIBF or in the median numbers when comparing ongoing vs. failed pregnancies. There was no B‐hCG interval where failed pregnancies were found to have lower frequency of PIBF expressing lymphocytes.

CONCLUSION: Inadequate PIBF expression independent of low P levels does not appear to be an etiologic factor for first trimester SABs; thus measuring this protein in pregnant women lacks practical usefulness.
A Study of Early Pregnancy Factor Activity in Preimplantation
Authors: Xiao‐Guang Fan , Zhen‐Qun Zheng , XIAO‐GUANG FAN , ZHEN‐QUN ZHENG
Published: September 2011

PROBLEM and METHOD: Early pregnancy factor (EPF), an Immunosuppressive substance, which appears in pregnant women's sera 48 h after fertilization, is a kind of pregnancy‐specific protein. To determine whether the EPF activity could be a super early indicator of pregnancy, we used rosette inhibition assay to detect EPF activity in the sera, collected from 70 women 2–7 days after ovulation intending to conceive monitored by ultrasonography. Simultaneously we selected 40 non‐pregnant sera and 12 early‐pregnant sera as negative control and positive control, respectively.

RESULTS: The results of this study demonstrated that EPF activity is detected in 35 women's sera out of 70 women within 2–7 days after ovulation, and 28 women out of the 35 were pregnant, which was known by follow‐up, and 7 were not pregnant, possibly due to either false positive results or embryo loss because of preimplantation failure, thus causing no pregnancy. The other 35 out of 70 had no EPF activity and 34 of them were not pregnant, which was known by follow‐up, but one case became pregnant, which was false negative result. Our study showed that diagnosis of the super early pregnancy could be made by detecting EPF activity in maternal serum within the time of preimplantation. The accuracy of pregnancy diagnosis by this method is 88.6%, with a false negative rate of 3.4% and a false positive rate of 17.1%. The β‐HCG level was measured from the above 70 women's sera in order to contrast EPF activity. All of the sera collected 2–6 days following ovulation indicated that there were lower β‐HCG values in very early pregnancy (≥a5 mIU/ml). On the seventh day after ovulation, EPF activity was detected in 11 out of 15 sera with only 2 of them with a b‐HCG level that reached or slightly surpassed that of the early pregnancy diagnosis (5 mIU/ml and 5.4 mIU/ml, respectively). This demonstrated that β‐HCG is not the earliest signal of pregnancy; otherwise the EPF activity is one that appears 2–6 days earlier than β‐HCG appears.

We measured the progesterone level of the 48 sera from the 70 collected above within 2–7 days postovulation and found most of them reached the level of progesterone in the luteal phase (7.5–98.3 nmol/L). This indicated that ovulation had taken place in these women, which was in accordance with observations by ultrasonography.

CONCLUSIONS: Our study showed that diagnosis (of 88.6%) of super early pregnancy could be made with an accuracy of 88.6% by detecting EPF activity in maternal serum within 2‐days after ovulation. This offers a basis for pregnancy diagnosis for the women who attempt to terminate their pregnancy safely or who conceive unexpectedly, and it contributes to family‐planning.
Distribution of Type‐I Interferon‐Receptors in Human First Trimester and Term Placental Tissues and on Isolated Trophoblast Cells
Authors: Paulesu Luana , Romagnoli Roberta , Fortino Vittoria , Cintorino Marcella , Bischof Paul
Published: September 2011

PROBLEM: Type‐I interferon (IFN) is the protein recognizing pregnancy in ruminants. Although IFN is secreted in early pregnancy, its role is not still clear in other species. Like other cytokines, IFN exerts its biological functions through specific membrane receptors. We have investigated the potential action of IFN in human pregnancy by studying the distribution of the receptors in the human placenta.

METHOD: Reactivity to monoclonal antibodies (mAbs) to the type‐I IFN‐receptor (R) was analyzed by immunohistochemistry in human placental tissues and in cytospins of first trimester trophoblast cells.

RESULTS: Type‐I IFN‐R immunoreactivity was observed mostly in first trimester villous cytotrophoblasts and in the cytotrophoblast cell columns. Trophoblast in the decidua, the epithelium of the uterine glands, and most of the isolated trophoblast cells were also immunoreactive.

CONCLUSION: The expression of type‐I IFN‐R in the highly proliferating and migrating trophoblast suggests that this cytokine has a role in trophoblast growth and invasion.
Histochemical Evaluation of Placental Dipeptidyl Peptidase IV (CD26) in Pre‐eclampsia: Enzyme Activity in Villous Trophoblast Indicates an Enhanced Likelihood of Gestational Hypertensive Disorders
Authors: Neudeck Hermann , Joncic Marion , Schuster Christof , Bisson Susanne , Hildebrandt Ruth , Öney Taylan , Stiemer Beate , Hopp Hartmut , Graf Renate
Published: September 2011

PROBLEM: To determine whether differences in placental dipeptidyl peptidase IV (DPP IV, CD26) activities occurred in hypertensive complicated pregnancies as compared with uncomplicated pregnancies.

METHOD: DPP IV activity was detected with H‐Gly‐Pro‐4M2NA as the substrate in placental cryostat sections from 65 patients with gestational hypertension and 67 patients with uncomplicated pregnancies. The graduated intensities of the reaction product in the villous trophoblast were scored semiquantitatively by light microscopy and were related to the relative frequencies of hypertensive disorders (proportional odds model). After detection of enzyme activity, the same tissue samples were homogenized and used for kinetic fluorometric measurements.

RESULTS: Enhanced villous trophoblastic DPP IV activity was significantly associated with an increased frequency of proteinuric hypertension in pregnant women (cumulative odds ratio θ1 = 1.6; P < 0.01).

CONCLUSION: This study demonstrates for the first time that increased villous trophoblastic DPP IV activity indicates an increased likelihood of the presence and of the severity of hypertensive disorders in pregnancy.
Induced Termination of Pregnancy by Purified Extracts of Azadirachta Indica (Neem): Mechanisms Involved
Authors: Talwar G.P. , Shah S. , Mukherjee S. , Chabra R.
Published: September 2011

PROBLEM: To develop a self‐administered, orally delivered method for abrogation of early pregnancy.

METHOD: Use of purified Neem extracts containing immunomodulators stimulating Th1 cells and macrophages; test animals, rats, baboons, and monkeys, onset of pregnancy confirmed by surgery and counting of implants on day 7 in rats and by chorionic gonadotropin (CG) and progesterone assays in primates; termination defined by complete resorption on day 15 in rats and by bleeding and decline of CG and progesterone in baboons.

RESULTS: Pregnancy was terminated successfully in both rodents and primates with no significant side effects. Fertility was regained in both species after one or two irregular cycles. Progeny born had normal developmental landmarks and mothered normal litters in the course of time. The active principle in Neem has been partially fractionated by activity‐guided purification. A cascade of events are involved in abrogation of pregnancy. In primates, a decrease in progesterone is an early event. A transient increase in CD4 and CD8 cells is noted in spleen at 96 hr and in mostly CD8 cells in mesenteric lymph nodes. Treatment causes an elevation of both immunoreactive and bioactive TNF‐α and γ‐interferon in serum, mesenteric lymph nodes, and foetoplacental tissue.

CONCLUSION: Immunomodulators of plant origin are potentially usable for termination of unwanted pregnancy
Evidence that the Expression of Progesterone‐Induced Blocking Factor by Maternal T‐Lymphocytes Is Positively Correlated with Conception
Authors: Check Jerome H. , Arwitz Maria , Gross Jenifer , Szekeres‐Bartho Julia , Wu Chung H.
Published: September 2011

PROBLEM: To compare the expression by T‐lymphocytes of an immunomodulatory protein known as progesterone‐induced blocking factor (PIBF) in conception versus non‐conception cycles even when there has been definite fertilization and embryo formation.

METHOD: PIBF expression on T lymphocytes was measured using an immunohistochemical method with a PIBF‐specific polyclonal antibody. These levels were determined in patients undergoing three types of therapy: non‐in vitro fertilization (IVF), IVF‐embryo transfer (ET), and frozen ET. Sera were drawn 12 days from ovulation in non‐IVF cycles or 9 days after ET and were assayed for PIBF and beta human chorionic gonadotropin. Comparison of the frequency of lymphocyte expression of PIBF in pregnant versus non‐pregnant women were made.

RESULTS: PIBF was detected in 29.5% of non‐pregnant women and 52.5% of pregnant women. There were no differences in PIBF levels by therapy used in non‐pregnant cases or in the pregnant group.

CONCLUSION: These data are consistent with the hypothesis that maternal expression of PIBF in T‐lymphocytes soon after trophoblast invasion may depend on successful implantation.
Immunomagnetic Isolation of Fetal Rat Gonocytes
Authors: van den Ham R. , van Pelt A.M.M. , de Miguel M.P. , van Kooten P.J.S. , Walther N. , van Dissel‐Emiliani F.M.F
Published: September 2011

PROBLEM: An efficient method to obtain highly enriched populations of viable gonocytes from rat embryos at Day 18 and Day 20 postcoïtum (pc) is described.

METHOD: Single‐cell suspensions with high cell yield were obtained by a collagenase/ trypsin digestion of the decapsulated testis. The gonocytes were purified by a direct immunoseparation technique,1,3 using magnetizable beads coated with rat anti‐mouse immunoglobulin M (IgM) and a monoclonal antibody 4B6.3E10, which specifically reacted with a differentiation antigen on the fetal germ cells.

RESULTS: Populations of 8.3 ± 2.7 (x 103; 18 days pc) or 1.2 ± 0.25 (x 104; 20 days pc) viable gonocytes per testis with purities of 91 ± 6.5% and 92 ± 4.3%, respectively, as determined by Nomarski microscopy were obtained.

CONCLUSION: The cells were successfully used for culture studies and as starting material for the investigation of gene expression.
Estradiol Down‐Regulates LPS‐Induced Cytokine Production and NFkB Activation in Murine Macrophages
Authors: Deshpande Rohini , Khalili Houman , Pergolizzi Robert G. , Michael Sandra D. , Chang Ming‐Der Y.
Published: September 2011

PROBLEM: In vivo and in vitro studies have indicated that estradiol can affect cytokine production in different cell types. This study examines whether estradiol affects inflammatory cytokine production by murine splenic macrophages.

METHODS: Mouse splenic macrophages were first treated with 17β‐estradiol, followed by lipopolysaccharide (LPS) stimulation. The production of cytokines by macrophages with or without estradiol treatment was determined at both the protein and mRNA levels. The nuclear factor‐kB (NFkB) activity of activated mouse splenic macrophages was also evaluated by electrophoretic mobility shift assay.

RESULT: Our results show that 17β‐estradiol decreases LPS‐induced IL‐1α, IL‐6, and TNF‐α production but not IL‐10, IL‐12, and macrophage inflammatory protein (MIP) production by splenic macrophages. Furthermore, inhibition of cytokine production by 17β‐estradiol was associated with a decreased LPS‐induced NFkB‐binding activity.

CONCLUSION: Because cytokines are important mediators of immune function, the alteration of cytokine production by 17β‐estradiol may thus have a profound effect on the outcome of immune response during inflammation.
Evidence for an Elevation in Serum Interleukin‐2 and Tumor Necrosis Factor‐α Levels Before the Clinical Manifestations of Preeclampsia
Authors: Hamai Yoko , Fujii Tomoyuki , Yamashita Takahiro , Nishina Hidenori , Kozuma Shiro , Mikami Yasuyo , Taketani Yuji
Published: September 2011

PROBLEM: The purpose of this study is to clarify whether the disruption of immune regulation occurs in early pregnancy before the clinical manifestations of preeclampsia.

METHOD OF STUDY: The serum concentrations of interleukin‐2 (IL‐2) and tumor necrosis factor‐α (TNF‐α) were determined by using enzyme‐linked immunoadsorbent assay (ELISA) in the first trimester of pregnancy in women who had preeclampsia develop after 28 weeks of pregnancy (preeclamptic group) and in women who completed pregnancy uneventfully (control group).

RESULTS: Serum concentrations of both IL‐2 and TNF‐α in the first trimester of the preeclamptic group were significantly higher than those of the control group.

CONCLUSIONS: That the perturbation of feto‐maternal immune regulation may precede the clinical manifestations of preeclampsia, which may be of relevance in the development of preeclampsia, is suggested.
Quantitative Analysis of Constitutive Heterochromatin in Couples with Fetal Wastage
Authors: Buretic‐Tomljanovic A. , Badovinac A. Radojcic , Vlastelic I. , Randic L.J.
Published: September 2011

PROBLEM: Heteromorphism of constitutive heterochromatin is a stable evolutionary feature that is thought to cause no phenotypic alterations. Nevertheless, the role of constitutive heterochromatin is still unknown. The instability of constitutive heterochromatin was generally restricted to T‐lymphocytes and was associated with variable immunodeficiency. The heterochromatin regions of chromosomes 1, 9, 16, and Y have been postulated to play a role in the immune response and during early embryo development.

METHOD OF STUDY: To investigate a possible influence of constitutive heterochromatin in human reproductive ability, quantitative analysis of constitutive heterochromatin in human chromosomes 1, 9, 16, and Y was done. Thirty couples were divided into two groups, owing to the clinical heterogeneity of their reproductive disorders. The first group included couples with two or more spontaneous abortions as the only pregnancy outcomes, and the second group included couples with a stillborn child with or without malformations. In the control group were couples with one or more healthy children without a history of fetal wastage. All of the persons in this study had normal karyotypes. The amount of constitutive heterochromatin was expressed by relative value using the simple transformation [q/(p + q)]. This value, obtained on GTG‐banded metaphase chromosomes, represented an indirect measure of heterochromatin content. The Y/F index was used to express the relative amount of heterochromatin in chromosome Y.

RESULTS: There was a significant increase in the heterochromatin content of the chromosome 16 homologue pair in males and females with a stillborn or a stillborn malformed child (P < 0.01) and an increase in total heterochromatin cell content compared to controls (P = 0.005). The same couples had significantly increased mean maximal heterochromatin content in the potential zygotes (P < 0.02). The couples who experienced spontaneous abortions only had a minimal total heterochromatin content in the potential zygotes (P < 0.05). The Y/F index was significantly lower in the males in both groups compared to controls (P1 < 0.02; P2 < 0.02).

CONCLUSION: The quantitative analysis of constitutive heterochromatin could be valuable in predicting pregnancy outcome.
Antisperm Antibody Detection: 2. Clinical, Biological, and Statistical Correlation Between Methods
Authors: Lenzi Andrea , Gandini Loredana , Lombardo Francesco , Rago Rocco , Paoli Donatella , Dondero Franco
Published: September 2011

PROBLEM: In clinical andrology, the detection of antisperm antibodies (ASA) is regarded as one of the most important steps in the study of male infertility. This practice is generally accepted even though there is still some disagreement about the true meaning of antisperm immunity, and there remains a good deal of controversy about the test regarded as the most suitable for the detection of antibodies directed against sperm antigens. International Workshops have tried to standardize universally accepted protocols. A panel of three or four methods is generally advised to provide a correct and complete screening of patients with antisperm immunity.

The aim of the present paper is to report our studies on the correlation between direct methods (IBT, MAR test) and indirect methods (gelatin agglutination test [GAT], and tray agglutination test [TAT]) and to establish whether biological models can explain the antibody tests results. An attempt was also made to establish a “predictive threshold” to explain even apparently discordant direct and indirect results.
Ten‐Year Experience with Antispermatozoal Activity in Ovulatory Cervical Mucus and Local Hydrocortisone Treatment
Authors: Ulčová‐Gallová Zdenka
Published: September 2011

PROBLEM: During 1987–1996, cervical mucus spermagglutinating antibodies of secretory immunoglobulin A (sIgA) or IgG detected by the tray agglutination test (TAT) and the indirect mixed antiglobulin reaction (MAR) test were found in 234 women aged 23–39 years with previously unexplained infertility.

METHOD OF STUDY: Hydrocortisone was applied to the ectocervix.

RESULTS: Spermagglutinating antibodies of sIgA disappeared totally in 102 patients (91 resulting in deliveries of healthy babies, 3 in ectopic pregnancies, and 8 in spontaneous miscarriages). A decrease of spermagglutinating antibodies in ovulatory mucus during hydrocortisone application without pregnancy was registered in 60 infertile women. This group was referred to treatment by in vitro fertilization. No hydrocortisone effect on immunocompetent cells producing antispermatozoal IgG alone or combined with sIgA was seen in 72 patients.

CONCLUSIONS: Vaginal mycosis, a side effect of hydrocortisone treatment, was seen in seven cases. Hydrocortisone for local immunosuppression becomes a valuable method of therapy in cervical immunologic infertility caused predominantly by antispermatozoal sIgA.
Flow Cytometric Analysis of Leukocytes in the Human Female Reproductive Tract: Comparison of Fallopian Tube, Uterus, Cervix, and Vagina
Authors: Givan Alice L. , White Hillary D. , Stern Judy E. , Colby Esther , Guyre Paul M. , Wira Charles R. , Gosselin Edmund J.
Published: September 2011

PROBLEM: The tissues of the human female reproductive tract (Fallopian tube, uterus, cervix, and vagina) may play different roles in the provision of mucosal immunity. The purpose of this study was to develop a uniform method suitable for quantitative comparison of the leukocytes from all these tissues.

METHOD OF STUDY: Tissues, typically 0.5‐1.0 g, were dispersed by enzyme treatment. A flow cytometric gating procedure based on CD45‐positivity and low far‐red autofluorescence permitted unfractionated, freshly dispersed cells to be phenotyped with respect to T lymphocytes, B lymphocytes, macrophages, and granulocytes.

RESULTS: Reproductive tract tissues contain leukocytes that represent approximately 6–20% of the total number of cells, with the Fallopian tubes and uterus containing a higher proportion of leukocytes than the cervix and vagina. The uterine endometrium from postmenopausal women has fewer leukocytes than does uterine endometrium from pre‐menopausal women. T lymphocytes are a major constituent (30–60%) of leukocytes from all tissues. The Fallopian tube contains granulocytes as another major constituent; granulocytes are significantly less numerous in the other tissues. All tissues contain B lymphocytes and macrophages as clearly detectable but minor components.

CONCLUSIONS: Three‐color flow cytometry is an appropriate method for quantitative comparison of leukocytes from the different tissues of the female reproductive tract, during all phases of the menstrual cycle and within post‐menopausal samples. Results indicate that the tissues differ from each other, particularly with respect to the large number of granulocytes in the Fallopian tubes.
Expression of IGFs and Their Receptors Is a Potential Marker for Embryo Quality
Authors: Liu Hung‐Ching , He Zhi‐Ying , Mele Carol A. , Veeck Lucinda L. , Davis Owen K. , Rosenwaks Zev
Published: September 2011

PROBLEM: Insulin‐like growth factors (IGFs) and insulin have been demonstrated to stimulate oocyte maturation and embryo development. Therefore, the expression of IGFs and their receptors may be an important intrinsic factor for embryo growth and may be a potential marker for embryo quality.

METHOD OF STUDY: Thirty donated day 3 embryos were cultured in vitro for an additional 3 days to observe their developmental potential and were semiquantitatively analyzed for the expression of IGF‐I, IGF‐II, IGF‐IR, IGF‐IIR, and insulin‐R.

RESULTS: Our results show that the activity of these gene expressions correlates well with the morphological assessment and that high and more gene expressions were often associated with embryos of high growth potential.

CONCLUSION: The IGF system may indeed play an important role in human embryogenesis; IGF gene expressions can be a good indicator of embryonic developmental stage and/or growth potential; finally, the IGF system can serve as a marker for embryo quality.
Immunolocalization of the Inducible Nitric Oxide Synthase Isoform in Human Fetal Membranes
Authors: Eis A.L.W. , Brockman D.E. , Myatt L.
Published: September 2011

PROBLEM: Nitric oxide (NO) synthesized by fetal membranes may protect the fetus from maternal infection or immune challenge or have a tocolytic effect on myometrium. The sites of synthesis and enzymes responsible for NO production in human fetal membranes remain unidentified.

METHOD OF STUDY: Fetal membranes were obtained from four groups of patients: term (>37 weeks gestation) or preterm (<37 weeks gestation), both either in labor or not in labor. Frozen sections of membrane rolls were immunostained for inducible (iNOS) and endothelial (eNOS) nitric oxide synthase isoforms and the monocyte/macrophage marker CD14.

RESULTS: Positive iNOS immunostaining was found in fibroblasts of amnionic and chorionic mesenchyme and in decidual macrophages identified by CD14 from all four groups of tissues. No iNOS immunostaining was seen in amnion epithelium or chorion trophoblast. Very intense iNOS staining was seen with evidence of monocyte/macrophage invasion of membranes. eNOS immunostaining was only found in decidual vascular endothelium.

CONCLUSIONS: Constitutive expression of iNOS in decidual macrophages and fetal membrane fibroblasts may form an immune barrier against maternal insult. In chorioamnionitis, macrophage recruitment and NO expression may be part of the maternal immune response.
Invasion of Cytotrophoblastic (JEG‐3) Cells Is Up‐Regulated by Interleukin‐15 In Vitro
Authors: Zygmunt M. , Hahn D. , Kiesenbauer N. , Münstedt K. , Lang U.
Published: September 2011

PROBLEM: Trophoblast invasion into the uterus is controlled by many factors. Some cytokines (interleukin [IL]‐1, IL‐6, and IL‐10) have been shown previously to play an important role in placentation. The human placenta is an important source of IL‐15, although the cellular source of IL‐15 in the placenta has not yet been specified. IL‐15 influences cell adhesion and migration by redistributing adhesion molecules in lymphocytes and has been shown to have effects on endothelial cells and in some human tumors.

METHOD OF STUDY: To study the role of IL‐15 in trophoblast invasion, we investigated the effect of IL‐15 (concentrations, 1–10 ng/ml) in a trophoblast invasion model (JEG‐3 with matrigel‐coated filters). Cell invasion was assessed using matrigel‐coated filters and was expressed as the quotient of invading cells in comparison with the number of cells that had passed the control membrane. Cell migration was studied by examining the number of cells that had passed the filters without matrigel. Cell proliferation was quantified by a tetrazolium salt WST‐1 cleavage assay. Matrix metalloproteinase (MMP)‐1, MMP‐2, and MMP‐9 activities were measured by specific enzyme assays.

RESULTS: IL‐15 significantly (P < 0.05) increased the in vitro invasion of cytotrophoblastic (JEG‐3) cells in a dose‐dependent manner. There was a fourfold increase in the invasion at a concentration of 10 ng/ml of IL‐15. Migration also was increased by a factor of 2.3 (P < 0.05). Cell proliferation, however, remained unchanged. The collagenolytic activity of cytotrophoblastic (JEG‐3) cells was increased by IL‐15 stimulation. A significant increase in MMP‐1 concentration occurred after the incubation of JEG‐3 cells with IL‐15. No changes appeared in MMP‐2, MMP‐9, and tissue inhibitor of metalloproteinase‐1 concentrations.

CONCLUSIONS: Trophoblast invasion and migration, but not proliferation, are enhanced by IL‐15. Our results suggest a role for IL‐15 in the modulation of MMP‐1 secretion by JEG‐3 cells. Furthermore, we speculate, that IL‐15 might be related to the changes of cell adhesion molecule phenotype during the process of invasion.
Comparative Studies on the Effects of Interleukin‐4 and Interleukin‐13 on Cytokine and Prostaglandin E2 Production by Amnion‐derived WISH Cells
Authors: Keelan Jeffrey A. , Sato Timothy A. , Mitchell Murray D.
Published: September 2011

PROBLEM: In hematopoietic cells, interleukin (IL)‐13 shares many actions with IL‐4. The effects of IL‐13 in gestational tissues have yet to be reported, however. We compared the effects of IL‐4 and IL‐13 on the production of cytokines and prostaglandin E2 (PGE2) in epithelial amnion‐derived WISH cells.

METHOD OF STUDY: WISH cells were treated with IL‐4 or IL‐13 (0.08–10 ng/ml) with/without cotreatment with IL‐1β (0.2 ng/ml), tumor necrosis factor‐α (10 ng/ml) or epidermal growth factor (5 ng/ml). The production of IL‐6, IL‐8, and PGE2 was measured by immunoassay after 16 hr.

RESULTS: Both IL‐4 and IL‐13 inhibited PGE2 production with indistinguishable concentration‐response curves, under basal or stimulated conditions. The maximal inhibition of IL‐1β‐stimulated PGE2 production (to 28% ± 10% of control) was seen at 10 ng/ml of IL‐4 or IL‐13. Basal IL‐6 production was stimulated approximately twofold by IL‐4 and IL‐13, whereas IL‐4 and IL‐13 both inhibited cytokine‐stimulated (but not basal) IL‐8 production by approximately 50%. In the presence of 1 ng/ml of IL‐4, IL‐13 was unable to further inhibit PGE2 production.

CONCLUSIONS: The inhibition of PGE2 and IL‐8 production by IL‐4 in WISH cells is mimicked by IL‐13. Both cytokines, probably through binding to a common receptor complex, may share a role in suppressing inflammatory reactions within intrauterine tissues.
Natural Killer Cell Cytotoxicity and Paternal Lymphocyte Immunization in Women with Recurrent Spontaneous Abortions
Authors: Kwak Joanne Y.H. , Gilman‐Sachs Alice , Moretti Michael , Beaman Kenneth D. , Beer Alan E.
Published: September 2011

PROBLEM: Natural killer (NK)‐cell cytotoxicity in women undergoing lymphocyte immunization prior to and following treatment was investigated.

METHOD OF STUDY: A cohort of 33 women with a history of two or more recurrent spontaneous abortions was prospectively studied. NK‐cell cytotoxicity was determined at effector‐to‐target ratios of 50:1 and 25:1. Peripheral blood CD56+ NK‐cell, CD 19+ B‐cell, CD19+/5+ B‐l‐cell, and CD3+ pan T‐cell levels were studied by flow cytometry before and after lymphocyte immunization treatment. Maternal antipaternal T‐ and B‐cell antibody levels were measured before and after lymphocyte immunization by flow cytometric analysis. Paternal lymphocyte immunizations were given two times with a 4‐week interval. Post‐lymphocyte immunization testing was done 4 weeks after the second lymphocyte immunization. The controls were 8 normal healthy women. NK assays were done twice with an interval of 8 weeks.

RESULTS: NK‐cell activity at effector‐to‐target ratios of 50:1 (P = 0.005) and 25:1 (P = 0.001) were significantly suppressed after lymphocyte immunization. CD3+ pan T‐cell levels after lymphocyte immunization were significantly increased compared with levels before lymphocyte immunization (P = 0.008). CD56+ NK‐cell levels were significantly suppressed after lymphocyte immunization (P = 0.016). There was no correlation between changes in NK cytotoxicity and differences in antipaternal lymphocyte antibody levels before or after lymphocyte immunization.

CONCLUSION: Lymphocyte immunization suppresses NK‐cell cytotoxicity and CD56+ NK‐cell levels and increases the peripheral blood CD3+ T‐cell population in women with recurrent spontaneous abortions.
Granulocyte Macrophage‐Colony Stimulating Factor Production by Autologous Endometrial Co‐Culture Is Associated with Outcome for In Vitro Fertilization Patients with a History of Multiple Implantation Failures
Authors: Spandorfer S.D. , Barmat L.I. , Liu H.‐C. , Mele C. , Veeck L. , Rosenwaks Z.
Published: September 2011

PROBLEM: To determine whether granulocyte macrophage (GM)‐colony stimulating factor (CSF) produced by autologous endometrial co‐culture was associated with outcome in 53 patients with a history of multiple in vitro fertilization failures.

METHOD OF STUDY: The conditioned media from endometrial co‐culture cells exposed or non‐exposed to human embryos was analyzed for GM‐CSF.

RESULTS: Exposure or non‐exposure to an embryo did not result in an enhancement of GM‐CSF production. Insignificant levels of GM‐CSF were determined from media alone. ROC analysis revealed that levels of GM‐CSF from supernatants of endometrial co‐culture exposed to embryos that measured below 130 pg/ml reflected a diminished prognosis (5/17 had a positive pregnancy vs. 21/36 with GM‐CSF levels greater than 130 pg/ml; P < 0.05).

CONCLUSIONS: The improved outcome associated with GM‐CSF values greater than 130 pg/ml may reflect: 1) a direct positive effect of GM‐CSF; 2) an embryotrophic factor upregulated by GM‐CSF; or, 3) that GM‐CSF functions as a marker for the importance of the glandular component in endometrial co‐culture systems.
Review: Analogies Between Trophoblastic and Malignant Cells
Authors: Mullen Catherine A.
Published: September 2011

PROBLEM: The question of how trophoblastic and malignant cells evade immunologic recognition and rejection by their host was studied. METHOD OF STUDY: A literature review was conducted.

RESULTS: Trophoblastic and malignant cells share a number of similarities. These include a lack of major histocompatibility complex antigen expression, resistance to lysis by natural killer cells, T‐helper cell‐2 (TH2)‐biased response, prostaglandin E production, and response to transforming growth factor beta. In addition, the analogies between trophoblastic and malignant cells extend into immunotherapy in which anti‐idiotype therapy has a viable role in the prevention of pregnancy loss and the treatment of cancer. CONCLUSIONS: Trophoblastic and malignant cells use a number of similar mechanisms to resist rejection by their host. By using similar strategies these cells are able to successfully co‐exist in an immunologically hostile environment.
The Role of Placental Trophoblast in the Pathophysiology of the Antiphospholipid Antibody Syndrome
Authors: Rote Neal S. , Vogt Elizabeth , Devere Gabrielle , Obringer Amy R. , Ng Ah‐Kau
Published: September 2011

PROBLEM: The antiphospholipid (aPL) antibody syndrome is characterized by severe pregnancy complications, the cause of which remains unknown. We hypothesized that the placental trophoblast is a target for aPLs.

METHOD OF STUDY: The effects of monoclonal aPLs on trophoblast function, including the invasion of JAR into matrigel‐coated filters and the effects of annexin V expression on BeWo, were investigated using choriocarcinoma models.

RESULTS: aPLs against phosphatidylserine (PS) significantly (P < 0.001) decreased the migration of JAR across the membrane. In the annexin V studies, undifferentiated BeWo did not express surface annexin V. After differentiation, BeWo expressed surface annexin V, which was removed in the presence of aPLs, resulting in increased binding of prothrombin.

CONCLUSIONS: PS is expressed on the trophoblast surface during differentiation and invasion of extracellular matrix. Our data suggest that aPLs against PS can directly affect trophoblast function by limiting the depth of decidual invasion and by concurrently creating a procoagulant surface on trophoblast exposed to the maternal circulation.
The Peritoneal Fluid Levels of Interleukin‐12 in Women with Endometriosis
Authors: Zeyneloglu Hulusi B. , Senturk Levent M. , Seli Emre , Bahtiyar Ozan M. , Olive David L. , Arici Aydin
Published: September 2011

PROBLEM: Interleukin‐12 (IL‐12) is produced mainly by monocytes/macrophages, and it induces proliferation and cytotoxicity of T‐cells and natural killer cells. In women with endometriosis, natural killer cell activity in the peritoneal fluid is significantly decreased. We aimed to measure the peritoneal fluid level of IL‐12 in endometriosis.

METHOD OF STUDY: We measured IL‐12 levels in peritoneal fluid samples from women with or without endometriosis and in supernatants from endometrial stromal, ovarian stromal, and mesothelial cell cultures, using a high‐sensitivity enzyme‐linked immunosorbent assay.

RESULTS: The median concentration of IL‐12 in the peritoneal fluid of women with endometriosis was 1.1 pg/ml (range, 0.2–5.5) and was 1.6 pg/ml (range, 0.4‐2.8) in women without endometriosis, not a statistically significant difference. IL‐12 was not detected in the supernatants of endometrial stromal, ovarian stromal, and mesothelial cell cultures. CONCLUSION: Concentrations of IL‐12 in the peritoneal fluid of women with or without endometriosis are low, but they are detectable and are not affected significantly by the presence of endometriosis.
Multivariant Analysis of Men from Infertile Couples With and Without Antisperm Antibodies
Authors: Gubin David A. , Dmochowski Roger , Kutteh William H.
Published: September 2011

PROBLEM: Research studies in animal and human systems have demonstrated conclusively that antisperm antibodies can interfere with fertilization. In the male, autoantibodies to sperm can be detected both in the sera and seminal plasma.

METHOD OF STUDY: Ninety‐seven men who were tested for antisperm antibodies as a part of an infertility evaluation were identified. Complete medical history was obtained, including information related to events suspected of being associated with antisperm antibodies. History of surgery (varicocele repair, hernia repair, and vas reversal) and infection (epididymitis, sexually transmitted disease, and orchitis) were compared with semen parameters (motility less than 60%, concentration less than 20 times 106, and volume less than 2 cc). These were compared to antisperm antibody results of mixed agglutination reaction (MAR) and direct immunobead binding test (IBT) for immunoglobulin G (IgG). Statistical analysis was performed using Fishers exact two‐tailed test.

RESULTS: As expected, prior vas reversal was significantly associated with the presence of antisperm antibodies (P = 0.0002) by MAR or IBT with a fivefold increased relative risk (95% confidence interval, 1.97‐12.38). Other surgeries manipulating the cord structures independent of vas reversal were not associated with antisperm antibodies (P = 0.09). Prior infections, independent of vas reversal, were significantly associated with antisperm antibodies by MAR (P = 0.04) with a 3.8‐fold increased relative risk (95% confidence interval, 1.06–13.87) but not by IBT. Sperm concentration less than 20 times 106, motility less than 60%, and a volume less than 2 cc were not associated with antisperm antibodies by MAR or IBT. CONCLUSION: These findings suggest that manipulation of the cord structures excluding the vas were not associated with antisperm antibodies; however, vas reversal and prior infection are significant risk factors for the development of antisperm antibodies.
Complement Regulatory Proteins on the Sperm Surface: Relevance to Sperm Motility
Authors: Jiang Haixiang , Pillai Subbi
Published: September 2011

PROBLEM: To determine whether complement regulatory proteins are present on human spermatozoa and whether antibodies to these proteins adversely affect sperm motility.

METHOD OF STUDY: Human sperm membrane proteins were solubilized and subjected to Polyacrylamide gel electrophoresis followed by Western blot analysis against antibodies to complement component 1 inhibitor (C1‐INH), decay‐activating factor (DAF; CD55), membrane cofactor protein (MCP; CD46), and homologous restriction factor (HRF; CD59). Spermatozoa, obtained by a swim‐up technique, were incubated in medium (control 1) and medium supplemented with antibodies to human albumin (control 2) and antibodies to these complement regulatory proteins. We used a computerized sperm motion analysis to determine the effect of these antibodies on sperm motion characteristics.

RESULTS: Complement regulatory proteins such as C1‐INH, CD55, CD46, and CD59 were found in the sperm extracts as shown by Western blot analysis. The heat‐treated (56°C, 30 min) IgG fraction of antibodies to these proteins significantly reduced sperm motility in general and other motion parameters. Addition of complement did not affect these results except in the antibodies to CD46 in which the reducing action was further amplified.

CONCLUSIONS: Our data suggest that C1‐INH, CD55, CD46, and CD59 are present on the sperm surface. These proteins may have biological functions, such as affecting sperm motility, besides the complement regulatory functions. In infertile men and women with antibodies that recognize one or more of these complement regulatory proteins, there may be problems related to poor sperm motility and survival in the reproductive tracts.
Identification of a Unique Form of p53 in Human Cord Blood Associated with the Development of Maternal Autoantibodies
Authors: Yashar Catheryn M. , Gercel‐Taylor Cicek , Gibb Randall K. , Weeks Jonathan W. , Taylor Douglas D.
Published: September 2011

PROBLEM: The possible link between p53‐reactive antibodies in multiparous women and exposure to a unique p53 protein during pregnancy was examined.

METHOD OF STUDY: p53‐reactive antibodies were evaluated in sera from nulligravid and multiparous women and patients with ovarian cancer by Western immunoblot. Furthermore, the presence of p53 protein was assayed in cord blood by enzyme‐linked immunosorbent assay. Cord blood‐derived p53 was compared structurally by protein fingerprinting and functionally by gel mobility shift assay to other isolates of p53.

RESULTS: Antibodies reactive with wild‐type p53 were observed in 92% of multiparous women and 42% were reactive with one tumor‐derived p53 protein. p53 protein was detected in 27 of 154 samples of cord blood. Structural analysis indicated that the fetal p53 resembled the UL‐1 p53. Functionally, the fetal and UL‐1 proteins failed to bind DNA.

CONCLUSIONS: Fetal p53 protein seems to be distinct from wild‐type p53, characterized by enhanced stability, structural differences and inability to bind DNA, analogous to alternatively spliced variants. Exposure to fetal p53 protein may form the basis for immunologic protection against cancer induced by multiparity.
Gonadotropins Induce the Release of Interleukin‐1β, Interleukin‐6 and Tumor Necrosis Factor‐α from the Human Preovulatory Follicle
Authors: de Mola J. Ricardo Loret , Goldfarb James M. , Hecht Bryan R. , Babbo C. Jan , Baumgardner Gavin P. , Friedlander Miriam A.
Published: September 2011

PROBLEM; The effects of exogenous gonadotropin administration and steroid levels on the release of various cytokines into the human follicular fluid (FF) were studied.

METHOD OF STUDY: Forty patients were included in two groups, those undergoing controlled ovarian hyperstimulation (COH) (n = 33) and natural cycles (n = 7). FF transvaginal aspirations were performed 36 hr after administration of human chorionic gonadotropin or a spontaneous surge of luteinizing hormone, respectively. FF cytokine measurements were performed with sensitive immunoassays.

RESULTS: FF cytokine levels were higher after COH [interleukin (IL)‐1β, 6.6 ± 0.32 pg/ml; IL‐6, 18.7 ± 2.1 pg/ml; and tumor necrosis factor (TNF)‐α, 32.5 ± 4.9 pg/ml] than in natural unstimulated cycles (0.52 ± 0.1 pg/ml, P < 0.001; 8.9 ± 1.2 pg/ml, P < 0.01; and 13.2 ± 2.6 pg/ml, P < 0.001, respectively). FF estradiol (E2) and progesterone levels were not statistically different between groups, despite the higher serum E2 levels observed in patients after COH.

CONCLUSIONS: Gonadotropins might regulate ovarian secretion of cytokines, because FF IL‐1β, IL‐6, and TNF‐α levels after COH were higher than during natural cycles.
Immunodetection of Cell Adhesion Molecules in Rat Sertoli Cell Cultures
Authors: Lustig Livia , Casanova Marta B. , Vianello Sergio E. , Denduchis Berta , Meroni Silvina , Cigorraga Selva
Published: September 2011

PROBLEM: The presence of cell adhesion molecules (CAMs) in Sertoli cells has not been explored extensively. The expression of CAMs involved in cell‐matrix and cell‐to‐cell interactions in Sertoli cell cultures was examined.

METHOD OF STUDY: Immunohistochemical and Western blot techniques were applied to rat Sertoli cell cultures using specific antibodies to α3, α5, and α6 integrin subunits; NCAM; and Cadherins.

RESULTS: Expression of α3 and α6 integrin subunits (mainly laminin receptors) and lack of expression of α5 integrin subunit (fibronectin receptor) was observed in Sertoli cells by immunohistochemistry. These cells also expressed neural CAM (NCAM) and N‐cadherin. By Western blot analysis, Sertoli cell extracts reacted with antibodies to α3 integrin subunit revealed a band approximately 130 kDa, whereas no expression of α5 integrin subunit was detected. Cell extracts incubated with antibodies to pan Cadherin exhibited a band approximately 120 kDa, whereas bands of 180, 140, and 120 kDa were observed with antibodies to NCAM.

CONCLUSION: New data about the expression of receptors for extracellular matrix proteins (α3 and α6 integrin subunits) as well as cell‐to‐cell adhesion molecules (NCAM and Cadherins) are reported in rat Sertoli cell cultures.
Cell‐Mediated Immunity to Human and Escherichia coli 60‐kDa Heat Shock Protein in Women: Association with A History of Spontaneous Abortion and Endometriosis
Authors: KLIGMAN ISAAC , JEREMIAS JAN , ROSENWAKS ZEV , WITKIN STEVEN S.
Published: September 2011

PROBLEM: Heat shock proteins are expressed during early pregnancy and in peritoneal fluids from women with endometriosis. The relationship between a cell‐mediated immune response to human 60‐kDa heat shock protein (hsp60), spontaneous abortion, and endometriosis was examined.

METHOD OF STUDY: Peripheral blood mononuclear cells (PBMCs) from 110 female partners of infertile couples undergoing in vitro fertilization and 41 fertile control subjects were incubated with human hsp60 or Escherichia coli hsp60. PBMC proliferation was measured by [3H]thymidine incorporation, and a stimulation index was calculated.

RESULTS: Lymphocytes from 21.8% of the infertile women, as opposed to 7.3% of the fertile women, proliferated in response to human hsp60 (P = 0.05). In contrast, proliferation in response to the E. coli hsp60 was equivalent in both groups. Within the infertile group, the response to human hsp60 was 40.7% among women with a history of spontaneous abortion and only 12% in those with no history of spontaneous abortion (P = 0.003). There was no association between immunity to E. coli hsp60 and spontaneous abortion or between immunity to human hsp60 and therapeutic abortion or the cause of infertility. Immunity to the E. coli hsp60 was associated with endometriosis.

CONCLUSIONS: A cell‐mediated autoimmune response to human hsp60 is associated with a history of spontaneous abortion, whereas immunity to E. coli hsp60 was most prevalent in women with endometriosis.