bluuebunny/crossref_metadata_2025 huggingface dataset:
Amniotic Fluid Granulocyte Colony Stimulating Factor Levels in Chorioamnionitis Do Not Predict Neonatal Sepsis
Authors: Hoskins Iffath Abbasi , Schatz Frederick , Zandieh Peyman , Lee Christina
Published: September 2011

PROBLEM: To assess the usefulness of amniotic fluid (AF) granulocyte colony‐stimulating factor levels (G‐CSF) in chorioamnionitis (CAM) to predict neonatal sepsis.

METHOD OF STUDY: AF samples were obtained from term and preterm patients with (Group I) and without (Group II) CAM and were assayed for G‐CSF levels. Patients with other infections were excluded. All AF samples were also tested for gram stain and cultures. The sensitivity, specificity, and predictive values of these parameters for diagnosing neonatal sepsis were assessed.

RESULTS: Positive AF cultures were the best predictors of neonatal sepsis in CAM, with a sensitivity of 67% and a positive predictive value (PPV) of 80%. Elevated AF G‐CSF levels (>1,000 pg/ml) were poor predictors of neonatal sepsis with a sensitivity of 29% and PPV of 39%.

CONCLUSION: Even though AF G‐CSF levels were markedly elevated in patients with CAM, they were poor predictors of subsequent neonatal sepsis.
Identification of the Source of Inhibins at the Time of Conception Provides a Diagnostic Role for Them in Very Early Pregnancy
Authors: Lockwood G.M. , Ledger W.L. , Barlow D.H. , Groome N.P. , Muttukrishna S.
Published: September 2011

PROBLEM: Early diagnosis of a complicated or poor pregnancy outcome in patients undergoing assisted reproductive technique (ART) fertility treatment could aid their counseling and management. A possible role for the inhibin superfamily as markers of early pregnancy viability was investigated.

METHOD OF STUDY: To determine the source of the dimeric glycoproteins inhibin A (α‐βA) in early pregnancy, serial blood samples from women who became pregnant following in vitro fertilization (IVF) with fresh embryo transfer (n = 50), from women who achieved pregnancy with frozen‐thawed embryos (n = 8), and from a control group of women with spontaneous conceptions (n = 7) were analyzed using a two‐site enzyme‐linked immunosorbent assay (ELISA). Gonadotropin‐releasing hormone (GnRH) analogues are routinely used in ART treatment cycles and are recognized to be luteolytic, and hence, periconceptual administration may be deleterious to pregnancy outcome. Serum samples were obtained from 8 IVF patients who conceived during the cycle in which they had inadvertent luteal phase exposure to GnRH analogues.

RESULTS: Elevated serum levels of inhibin A were detected in ongoing pregnancies from 4 weeks' gestation and increased to an initial peak at 9–10 weeks' gestation. Significantly higher levels (P < 0.05) were found in the multiple pregnancies, and nonviable clinical pregnancies had very low levels of inhibin A. Inhibin pro‐αC was detectable at levels above normal late luteal values in singleton and multiple pregnancies arising from IVF with fresh embryo transfer. In pregnancies established without corpus luteum activity, frozen‐thaw embryo replacement, the levels of pro‐αC containing inhibins were extremely low, suggesting that the corpus luteum is the major source of the alpha monomer. In pregnancies following inadvertent periconceptual exposure to GnRH analogue, the levels of pro‐αC were statistically significantly higher in successful pregnancies than in early pregnancy failures.

CONCLUSIONS: The feto‐placental unit is confirmed as the major source of inhibin A in early pregnancy, and the initially low levels and very rapid decline in inhibin A in pregnancies with embryonic failure suggest a role for this glycoprotein as a monitor of early pregnancy viability. The corpus luteum is demonstrated to be the major source of inhibin pro‐αC in early pregnancy, and very low levels in patients with peri‐implantational exposure are indicative of lytic damage and herald pregnancy failure despite luteal supplementation with progesterone.
Anti‐inflammatory Effects of Interleukin‐4, Interleukin‐10, and Transforming Growth Factor‐β on Human Placental Cells In Vitro
Authors: Goodwin V.J. , Sato T.A. , Mitchell M.D. , Keelan J.A.
Published: September 2011

PROBLEM: To determine the effects of anti‐inflammatory cytokines on the production of inflammatory mediators by placental cells.

METHOD OF STUDY: Cells from term human placentas were isolated and cultured in vitro in the presence of anti‐inflammatory cytokines interleukin (IL)‐4, IL‐10, and transforming growth factor (TGF)‐β. Their effects on the production of IL‐8 and prostaglandin E2 (PGE2) were investigated under basal conditions and after stimulation with IL‐1β and tumor necrosis factor (TNF)‐α.

RESULTS: Both IL‐10 and IL‐4 inhibited IL‐1β‐ and TNF‐α‐induced PGE2 production but had no significant effects on the production of PGE2 under basal conditions. TGF‐β1 was without effect in stimulated cells, whereas under basal conditions TGF‐β1 stimulated PGE2 production. Similar trends were seen for IL‐8 production, with the exceptions that TGF‐β1 decreased the TNF‐α‐induced production and IL‐4 decreased basal IL‐8 production.

CONCLUSIONS: The anti‐inflammatory effects shown by IL‐4, IL‐10, and (to lesser extent) TGF‐β may play a role in ameliorating the potentially harmful effects of pro‐inflammatory mediators in the feto‐placental unit.
Gestational Age‐Dependent Extravillous Cytotrophoblast Osteopontin Immunolocalization Differentiates Between Normal and Preeclamptic Pregnancies
Authors: Gabinskaya Tatyana , Salafia Carolyn M. , Gulle Veronica E. , Holzman Ian R. , Weintraub Andrea S.
Published: September 2011

PROBLEM: Normal placentation requires modulation of proliferative cytotrophoblast to an invasive phenotype. Preeclampsia is characterized by failed cytotrophoblast invasion and arterial remodeling. Osteopontin (OPN) is an extracellular matrix protein implicated in cell adhesion, spreading, and invasion.

METHOD OF STUDY: To investigate gestational age‐related OPN expression, placental immunostaining was performed. To investigate the role of OPN in uteroplacental vascular pathology, placental immunostaining from pregnancies with preeclampsia (n = 12), fetal growth retardation (FGR) (n = 8), or both (n = 4) was compared with gestational age‐matched controls (n = 24).

RESULTS: In non‐preeclamptic pregnancies, OPN immunolocalized to basal plate and intervillous cytotrophoblasts from 24–30 weeks (n = 13). In preeclampsia, OPN immunoreactivity was detected from 24–40 weeks. Cytotrophoblasts from FGR placentas were OPN‐positive until 30 weeks, unless preeclampsia accompanied the FGR. In this case, cytotrophoblasts were OPN‐positive from 24–40 weeks.

CONCLUSIONS: The data suggest a role for OPN in cytotrophoblast invasion of the maternal vasculature/extracellular matrix during non‐preeclamptic placentation, and OPN may serve as a marker for placental bed remodeling.
Labor Affects Cytokine Production in Newborns
Authors: Bessler H. , Kuperman A. , Beilin B. , Klinger G. , Gurary N. , Mozes C. , Sirota L.
Published: September 2011

PROBLEM: To investigate whether the mode of delivery or the drugs given to the mother during labor may affect the newborns' immune system.

METHOD OF STUDY: Three groups of term newborns were included: A, spontaneously delivered with i.v. analgesia (n = 37); B, spontaneously delivered with epidural analgesia (n = 26); and C, delivered by cesarean section under general anesthesia (n = 29). Natural killer (NK) cell cytotoxicity, mitogenic response, and the capacity of peripheral blood mononuclear cells (PBMCs) to produce interleukin (IL)‐1β, IL‐2, IL‐3, IL‐6, and tumor necrosis factor (TNF)‐α were examined.

RESULTS: NK cell cytotoxicity increased significantly in all three groups of newborns on the second day of life. Decreased IL‐2 production was observed in newborns delivered by cesarean section. Spontaneous IL‐1β secretion was higher in newborns to mothers treated with epidural analgesia. Spontaneous IL‐6 secretion was elevated in infants to mothers undergoing general anesthesia and surgery or epidural analgesia. TNF‐α production was increased in newborns delivered by cesarean section.

CONCLUSION: The immune response of the newborn is affected by the mode of delivery and/or drugs given to the mother during labor.
Inhibin A Concentrations in the Sera of Young Women During and After Chemotherapy for Lymphoma: Correlation with Ovarian Toxicity
Authors: Blumenfeld Z. , Ritter M. , Shen‐Orr Z. , Shariki K. , Ben‐Shahar M. , Haim N.
Published: September 2011

PROBLEM: Inhibin A concentrations in serum may reflect the ovarian granulosa cell compartment. To characterize the correlation between ovarian function after gonadotoxic chemotherapy for Hodgkin's or non‐Hodgkin's lymphoma in young women, the immunoreactive inhibin A concentrations in the sera of these patients was measured before, during, and after the gonadotoxic chemotherapy.

METHOD OF STUDY: A prospective clinical protocol was undertaken in 20 cycling women with lymphoma, aged 15–40 years. A monthly injection of depot D‐TRP6 ‐GnRH‐a (Decapeptyl CR, Ferring) was administered from before starting the chemotherapy until its conclusion, up to a maximum of six monthly injections. Most of the patients were treated with the mustargen‐oncovin‐procarbazine‐prednisone (MOPP)/actinomycin D‐bleomycinvincristine (ABV) chemotherapy combination; 13 with and 7 without radiotherapy. A hormonal profile [follicle‐stimulating hormone (FSH), luteinizing hormone (LH), 17‐β‐estradiol (E2), testosterone (T), progesterone (P4), insulin‐like growth factor (IGF)‐1, IGF‐bp3, and prolactin (PRL)] was taken before starting the gonadotropin‐releasing hormone agonist (GnRH‐a)/chemotherapy co‐treatment and monthly thereafter until resuming spontaneous ovulation and menstrual cyclicity. This group of prospectively treated lymphoma patients was compared with a control group of 22 regularly cycling women who had been treated with chemotherapy (mostly MOPP/ABV) with or without radiotherapy for Hodgkin's or non‐Hodgkin's lymphoma. Inhibin A immunoactivity developed by Nigel Groome was measured by an enzyme‐linked immunoadsorbent assay (ELISA) commercial kit (Serotec).

RESULTS: Whereas all but one (40 years of age) of the surviving patients in the GnRH‐a/chemotherapy co‐treatment group resumed spontaneous ovulation and menses within 6 months, only one half of the patients in the “control” group (chemotherapy without GnRH‐a co‐treatment) resumed ovarian function and regular cyclic activity (P < 0.05). The remaining 50% experienced premature ovarian failure (POF). Temporarily increased FSH concentrations were experienced by approximately one third of the patients resuming cyclic ovarian function, suggesting a reversible ovarian damage in a larger proportion of women than those experiencing POF. The inhibin A immunoactive concentrations decreased during the GnRH‐a/chemotherapy co‐treatment but increased to normal levels in patients who resumed regular ovarian cyclicity, and/or spontaneously conceived, as compared to low levels in menopausal women and those who had developed POF.

CONCLUSIONS: If these preliminary data are consistent in a larger group of patients, inhibin A concentration may serve as a prognostic factor for predicting the resumption of ovarian function, in addition to the levels of FSH, LH, and E2.
Tumor Necrosis Factor‐α mRNA‐Positive Cells in Spontaneous Resorption in Rodents
Authors: Lea Richard G. , McIntyre Susan , Baird Joyce D. , Clark David A.
Published: September 2011

PROBLEM: It has been proposed that high rates of resorption/spontaneous abortion may result from interaction in the decidua of γ‐interferon‐producing natural killer (NK) cells and tumor necrosis factor (TNF)‐α‐producing macrophages. An increased release of TNF‐α from placental tissue of resorptions has been reported, but macrophages producing TNF‐α have so far not been demonstrated at the feto‐maternal interface. Therefore, we have sought to identify TNF‐α‐producing cells by in situ hybridization at the feto‐maternal interface in two inbred, well‐characterized, and stable strains of laboratory rodents with high and low resorption rates.

METHOD OF STUDY: Pregnant DBA/2‐mated CBA/J mice with a resorption rate of 20% to 30% (dependent on NK cells and macrophages) and diabetes‐resistant Bio‐Breeding/ Edinburgh (DR‐BB/E) rats with low resorption rates (presumed to result from chromosomal abnormalities) were studied. AsialoGMl+ cells were detected by immunohistochemistry, and TNF‐α mRNA+ cells were detected by in situ hybridization.

RESULTS: TNF‐α mRNA+ cells were detected in DBA/2‐mated CBA/J mice at the time of resorption but only at the trophoblast‐decidual junction. AsialoGM1+ cells were present in decidua, as assessed by immunohistochemistry, but few if any gave a positive signal for TNF‐α. In rat resorptions, TNF‐α mRNA‐positive cells were present within the yolk sac and in contact with the trophoblast, but not at the trophoblast‐decidual junction. In neither species did a significant accumulation of detectable TNF‐α mRNA+ cells occur before the usual time of onset of resorption.

CONCLUSIONS: In the DBA/2‐mated CBA/J mouse, the removal of the placenta is associated with removal of a thin rim of adherent decidua similar to the location of the TNF‐α mRNA+ cells detected in this study. Our data suggest that increased TNF‐α in tissues associated with failing feto‐placental units may arise from infiltration/activation of scavenger cells from decidua that are likely to be macrophages. Local TNF‐α production in decidua, which occurs as a prelude to resorption in the CBA x DBA/2 model, could not be detected due to the insensitivity of the TNF‐α probe we used; the release of TNF‐α from decidual tissue left after the removal of the placenta does not differ between resorbing and healthy implant sites. AsialoGM1+ cells did not seem to be major producers of TNF‐α. TNF‐α mRNA+ cells in a low rate of resorption (rat) model were only found on the fetal side of the trophoblast, and they may also represent a macrophage response (to dying embryo tissue) derived from a nondecidual source. The location of TNF‐α mRNA+ cells may identify distinct and different mechanisms of resorption in rodents.
Anti‐Endothelial Cell Antibodies: Another Cause for Pregnancy Loss?
Authors: Roussev Roumen G. , Stern Jaroslav J. , Kaider Brian D. , Thaler Christian J.
Published: September 2011

PROBLEM: Previous studies demonstrated that unique tissue‐specific antigens expressed on vascular endothelial cells can serve as immunogens, and the apparent association between transplant rejection and antiendothelial cell (EC) antibodies is well established. A common feature of some placentas from women with recurrent pregnancy loss is the diffuse formation of microthrombi associated with changes in thromboresistant properties of endothelial cells, similar to the findings in rejected organs. Therefore, the prevalence of anti‐EC antibodies in patients with recurrent pregnancy loss and the role of these antibodies in cultured human endothelial cells from umbilical cord vein were studied.

METHOD OF STUDY: To evaluate the frequency of anti‐EC antibodies, sera from 160 nonpregnant patients with recurrent pregnancy loss after absorption with pooled platelets and pooled leukocytes were tested using cultured human umbilical cord vein endothelial cells (HUVEC) by flow cytometry. To study the role of anti‐EC antibodies, purified anti‐EC IgGs were added to HUVEC cultures and hemostatic, fibrinolytic, and anticoagulation pathway markers (tissue factor, tissue plasminogen activator, palsminogen‐activator inhibitor, thrombomodulin, heparan‐sulfate proteoglican, antithrombin III, von Willebrand factor, CD54, human leukocyte antigens‐DR, and transferin receptor) were detected by specific antibodies and flow cytometry. Immunoblotting analyses were done by using purified anti‐EC IgGs against cell membrane proteins from endothelial cells and leukocytes extracted by detergent solubilization.

RESULTS: Thirty‐nine (24%) of the patients were positive for EC antibodies. Antipaternal lymphocyte antibodies were tested as well and were found in 37 (23%) of the patients as 25 sera (15.6%) showed reactivity with both EC and lymphocytes. Certain patient sera were reactive with HUVEC lines from some but not other umbilical cords, which suggests allotype Sera from 70 normal healthy male and nonpregnant female controls did not react with any of the individual HUVEC lines used. The purified anti‐EC immunoglobulin G (IgGs) recognized bands, from HUVEC surface membrane protein preparations, with molecular weights of 120 kDa. Both hemostatic and fibrinolytic pathway markers were found activated in the presence of anti‐EC IgGs, suggesting an altered endothelial cell surface activation state.

CONCLUSIONS: The results indicate that anti‐EC antibody is another marker for a subset of recurrent spontaneous aborters who may have activation of hemostasis and fibrinolysis as a mechanism involved in their losses.
The Production of MLR‐Blocking Factors After Lymphocyte Immunotherapy for RSA Does Not Predict the Outcome of Pregnancy
Authors: Peña Ruth Beatriz , Cadavid Angela P. , Botero Jorge H. , García Gloria P. , Gallego María I. , Ossa Jorge E.
Published: September 2011

PROBLEM: The questions of whether production of mixed lymphocyte reaction‐blocking factors (MLR‐BFs) after immunotherapy with lymphocytes for recurrent spontaneous abortion (RSA) has prognostic value and whether cytotoxic antibodies are also involved were tested.

METHOD OF STUDY: A prospective study with 33 patients who had a history of two or more abortions, lacking MLR‐BFs, was carried out. The patients received immunizations with lymphocytes and 6 weeks or later were tested for seroconversion of MLR‐BFs. Seventeen of these thirty‐three patients were evaluated for antipaternal cytotoxic antibodies. The results were correlated with the outcome of the next pregnancy after treatment. RESULTS: Eighty percent of the 33 patients had a live child. Of those patients having success, only 50% produced MLR‐BFs. Of those patients having a new loss, five did and two did not produce MLR‐BF (P > 0.05). Regarding the 17 patients tested for cytotoxic antibodies, 4 of the 5 patients who tested positive had a new abortion, whereas only 1 of 12 whose tests remained negative did not have gestational success (P < 0.01). CONCLUSION: The presence of MLR‐BFs is not a prognostic criterium for the outcome of pregnancy after alloimmunotherapy, and, consequently, it is not a good diagnostic tool for RSA of alloimmune cause.
Soluble Intercellular Adhesion Molecule‐1 Serum Profile in Physiologic and Preeclamptic Pregnancy
Authors: Airoldi Lucia , Gaffuri Barbara , Rossi Gabriele , Iurlaro Enrico , Nozza Arrigo , Vigano Paola , Vignali Mario
Published: September 2011

PROBLEM: The aim of this study was to determine serum levels of soluble intercellular adhesion molecule (sICAM)‐1, an adhesion receptor that mediates interactions with the immune system, in physiologic and preeclamptic pregnancies. Moreover, we evaluated whether the release of sICAM‐1 during pregnancy correlated to plasma fibronectin concentrations.

METHOD OF STUDY: Serum was collected from 18 nonpregnant, control women, from 58 normal pregnant women during the first (n = 13), second (n = 15), and third (n = 30) trimesters, and from 25 preeclamptic patients at 27–39 weeks' gestation. All samples were assayed for sICAM‐1 by a specific enzyme‐linked immunoassay and for fibronectin by a nephelometric system. Serum sICAM‐1 levels in preeclamptic patients were compared to those obtained from gestational‐matched normal pregnant women.

RESULTS: Levels of sICAM‐1 were significantly elevated (P < 0.001) in each of the three trimesters of normal pregnancy (I trimester: 390.4 ± 25.7 ng/ml; II trimester: 386.3 ± 15.4 ng/ml; and III trimester: 367.3 ± 15.8 ng/ml) when compared to those of healthy nonpregnant women (263.3 ± 11.6 ng/ml). No significant difference in sICAM‐1 concentrations was observed among the three trimesters. Preeclampsia was associated to a significant decrease (P < 0.01) of sICAM‐1 levels (309.8 ± 11.6 ng/ml) relative to those observed in gestational‐matched pregnant women (367.3 ± 15.8 ng/ml). Fibronectin and sICAM‐1 levels did not correlate.

CONCLUSION: The increased levels of sICAM‐1 found in physiologic pregnancies and its reduction in preeclampsia may account for some of the immunologic alterations demonstrated to be associated with pregnancy.
Vaginal Immunization with Recombinant Gram‐Positive Bacteria
Authors: Medaglini Donata , Oggioni Marco R. , Pozzi Gianni
Published: September 2011

PROBLEM: Many viral and bacterial pathogens enter the body through the genital mucosa. Therefore, one of the major goals of a vaccine against sexually transmitted diseases (STDs) should be to induce an immune response in the genital mucosa capable of controlling the entry of the pathogen. Our approach for the development of vaccines against STDs is based on the use of nonpathogenic Gram‐positive bacteria as live vaccine vectors.

METHOD OF STUDY: Recombinant Gram‐positive bacteria expressing vaccine antigens were constructed using genetic systems developed in our laboratory. Balb/c mice and Cynomolgus monkeys were inoculated by the vaginal route and vaginal samples were collected using absorbent wicks. Colonization was evaluated by the presence of recombinant bacteria in the vaginal samples. Local and systemic immune responses were studied.

RESULTS: We have developed genetic systems for the expression of heterologous antigens on the surface of the human commensals Streptococcus gordonii and Lactobacillus spp. Both S. gordonii and L. casei stably colonized the murine vagina after a single inoculum. Vaginal colonization of mice with recombinant strains of S. gordonii, expressing human papillomavirus (HPV) and human immunodeficiency virus (HIV) antigens, induced antigen‐specific vaginal immunoglobulin A (IgA) and serum IgG. Local and systemic immune responses also were detected in monkeys immunized intravaginally with recombinant S. gordonii.

CONCLUSION: The results obtained indicated that the approach of using colonizing Gram‐positive bacteria as live vectors has a great potential for the development of vaccines against STDs.
Neutralizing Antibodies and Complement‐Mediated, Antibody‐Dependent Enhancement (C'‐ADE) of Human Immunodeficiency Virus Infection in Its Vertical Transmission
Authors: Gras Gabriel , Beyssen Véronique , Tranchot‐Diallo Juliette , Dormont Dominique , Parnet‐Mathieu Françoise , Lasfargues Géraud , Courpotin Christian
Published: September 2011

PROBLEM: Mother‐to‐child transmission is a major route for the spread of human immunodeficiency virus (HIV) worldwide. Our understanding of its mechanisms and parameters is still limited. Among the factors possibly involved in virus passage determination are the level and quality of antiviral humoral response.

METHOD OF STUDY: Anti‐HIV‐1/Lai neutralizing activity in sera from 35 mother‐infant pairs (in which 13 transmission cases occurred) was investigated, as was the complement‐mediated antibody‐dependent enhancement capacity of the same sera.

RESULTS: Neutralization titers of 640 or more were found only in four mothers of uninfected children, but this result was not significant. No significant link was obtained with the occurrence of complement‐mediated, antibody‐dependent enhancement.

CONCLUSIONS: As suggested by a synthesis of the literature, vertical transmission of HIV is probably the result of multiple active and/or stochastic parameters in the mother, the fetal structures, and the viral population. The precise definition of cellular mechanisms involved in in utero infection would help to better define which immune activity in the mother should be more carefully considered.
Fas‐Fas Ligand System‐Induced Apoptosis in Human Placenta and Gestational Trophoblastic Disease
Authors: Mor Gil , Gutierrez Linda S. , Eliza Mariel , Kahyaoglu Ferahnaz , Arici Aydin
Published: September 2011

PROBLEM: The low frequency of maternal immune responses to paternally inherited fetal antigens raises the following question: What regulates the immunobiology of pregnancy? Data suggest that this state is the result of peripheral immune‐tolerance, an active process of immune‐regulation in which activated T cells undergo apoptosis. We studied Fas ligand (FasL) expression and apoptosis in normal and pathologic placentas to find out whether the Fas‐FasL‐induced apoptosis takes place during implantation.

METHOD OF STUDY: FasL expression in paraffin sections was detected using specific antibodies and confirmed with reverse transcriptase‐polymerase chain reaction of total RNA from frozen placentas. Apoptosis was detected using the terminal deoxy (d)‐UTP nick end‐labeling assay.

RESULTS: FasL was found in the normal placenta and in gestational trophoblastic disease. Apoptotic leukocytes were localized to the maternal‐fetal interface corresponding in localization with the distribution of FasL.

CONCLUSIONS: We propose that FasL expression in the placenta is a mechanism responsible for the development of maternal immune tolerance specific for paternal alloantigens and operates in pathologic states characterized by trophoblastic invasion/proliferation.
Circulating Levels of Immunoreactive Cytokines in Women with Preeclampsia
Authors: Conrad Kirk P. , Miles Theresa M. , Benyo Deborah Fairchild
Published: September 2011

PROBLEM: Circulating inflammatory cytokines have been implicated in the pathogenesis of preeclampsia. To test this hypothesis, we measured plasma levels of immunoreactive tumor necrosis factor (TNF)‐α and ‐β, interleukin (IL)−1α and ‐β, and IL‐6 and −10 in women with preeclampsia, in women with transient gestational hypertension, and throughout normal pregnancy.

METHOD OF STUDY: Enzyme‐linked immunosorbent assays were used and subjected to extensive validation studies.

RESULTS: The median concentration of plasma TNF‐α was increased by twofold in women with preeclampsia compared with that in normal third‐trimester pregnancy (P < 0.001) and in women with gestational hypertension (P < 0.04). The median concentration of plasma IL‐6 was increased by threefold in women with preeclampsia compared with that in normal third‐trimester pregnancy (P < 0.001) and increased twofold compared with that in women with gestational hypertension (P < 0.1). There were no significant differences observed in the levels of plasma IL‐1β and IL‐10 between the preeclamptic and other subject groups. The level of IL‐1β, but not the levels of IL‐10, TNF‐α, or IL‐6, was significantly changed during normal pregnancy compared with the nonpregnant condition manifesting an overall decline (P < 0.04). TNF‐β and IL‐1α were not detected in any samples, possibly because of the low sensitivity of these particular immunoassays.

CONCLUSION: Elevated levels of TNF‐α and IL‐6 may contribute to the putative endothelial dysfunction of preeclampsia.
Apoptosis of T Cells in the First Trimester Human Decidua
Authors: JERZAK MAIGORZATA , KASPRZYCKA MONIKA , WIERBICKI PIOTR , KOTARSKI JAN , GÓRSKI ANDRZEJ
Published: September 2011

PROBLEM: Apoptosis has been accepted as a mechanism for maintaining tolerance in the immune system. The induction of apoptotic cell death can also be a possible outcome of the lymphocyte activation. Expression of Fas ligand (FasL) by the human trophoblast has been proposed as a mechanism providing protection against the lytic action of decidual immune cells. The aim of this study was to determine whether decidual T cells undergo apoptosis during abortion.

METHOD OF STUDY: We studied apoptosis of T cells isolated from the first‐trimester decidua in 12 women after spontaneous or elective abortion. We used gel electrophoresis to detect DNA fragmentation. Cells undergoing DNA fragmentation also were identified by DNA analysis using flow cytometry. This method was based on the accumulation of ethanol‐fixed apoptotic cells in the sub‐GO/G1 peak of the DNA content as a result of the loss of DNA fragments from the cells and because of a reduced DNA ability to be stained by propidium iodide. In addition, the expression of Fas antigen on the surface of decidual T cells (CD3+) also was determined.

RESULTS: We did not detect apoptosis by the “ladder” technique. However, the apoptotic index (the percentage of positive cells per total number of cells) ranged from 2% to 24% using flow cytometry.

CONCLUSIONS: Trophoblast cells usually fail to stimulate alloantigen‐specific T cells, but they may express nonclassical major histocompatibility complex alloantigens to which mothers can produce immunoglobulin G alloantibody, which requires T helper cell activation. The apoptosis of T cells in the human decidua, probably through Fas‐FasL signaling, may be a defense mechanism against rejection of the fetal allograft by the maternal immune system.
Major Histocompatibility Complex Expression on Human, Male Germ Cells: A Review
Authors: FISZER DOROTA , KURPISZ MACIEJ
Published: September 2011

PROBLEM: The male reproductive compartment is an immunologically privileged site. The expression pattern of human leukocyte antigens (HLAs) may play an important role in the maintenance of immune tolerance toward differentiating gametogenic cells. This review presents current knowledge about HLA gene expression on human, male germ cells, on mRNA and protein levels, and on their methylation status.

METHOD OF STUDY: Different techniques were applied to study HLA gene expression in human testis: (a) protein: e.g., cytotoxicity test, fluorescent labeling techniques, enzyme‐linked immunosorbent assay, and confocal microscopy; (b) mRNA: reverse transcriptase‐polymerase chain reaction, Northern blot hybridization, and in situ hybridization; and (c) methylation status.

RESULTS: In normal testicular tissue we observe a lack of HLA‐class I (classical) antigens expression and inversely related expression pattern of HLA class I classical and nonclassical genes. HLA‐A, ‐B, ‐C, and ‐E loci are likewise methylated in somatic and germ cells, whereas ‐F and ‐G genes are less methylated in sperm precursors.

CONCLUSIONS: Immunologic tolerance in human testis is actively maintained by the specific expression pattern of HLA genes regulated by hormones and growth factors.
Sexual and Mother‐to‐Child Transmission of the Human Immunodeficiency Virus Type 1: A Review
Authors: CHERMANN JEAN‐CLAUDE
Published: September 2011

PROBLEM: Sexual and mother‐to‐child transmission of the human immunodeficiency virus (HIV) type 1 occurs only with a low percentage of infection. Many instances of sexual intercourse result in no transmission, and only 20% of children are infected from seropositive mothers (3% in mothers treated with azidothymidine).

METHOD OF STUDY: We analyzed the presence of HIV in various ejaculates of the same HIV‐infected patients, as well as in the cervico‐vaginal fluid. We have studied the mechanism of transmission from mother to child, by analyzing the cell‐to‐cell transmission in the trophoblast.

RESULTS: Some ejaculates collected at different times from the same HIV‐infected males are free of virus, explaining the low rate of sexual transmission. We never found HIV in mobile spermatozoa. The trophoblast can be infected by HIV with a strain dependence and also transiently. By analyzing the tissue of the fetus, it was found that only some organs are infected, confirming the cell‐to‐cell transmission between the mother and child and not a true vertical transmission through the germinal lines.

CONCLUSIONS: HIV is not always present in the genital secretion, explaining the low rate of sexual transmission. Mother‐to‐child transmission occurs during pregnancy but often after the second trimester and at delivery after cell‐to‐cell or blood transmission, respectively.
Epitopes of Human Chorionic Gonadotropin and Their Relationship to Immunogenicity and Cross‐Reactivity of β‐Chain Mutants
Authors: PORAKISHVILI N. , JACKSON A.M. , de SOUZA J.B. , CHIESA M. DALLA , ROITT I.M. , DELVES P.J. , LUND T.
Published: September 2011

PROBLEM: Human chorionic gonadotropin (hCG) is a placental glycoprotein hormone, a heterodimeric molecule, consisting of α and β chains. It induces the synthesis of progesterone, which is essential for the maintenance of the fertilized egg. Antibodies directed against hCG can, therefore, prevent pregnancy and serve as a vaccine. hCG belongs to the glycoprotein hormone family and shares the α chain with the other members. The β chain is a hormone‐specific subunit that is unique to hCG, but still possesses 85% amino acid homology with the β chain of luteinizing hormone (LH), which means that prolonged immunization with hCG produces antibodies that cross‐react with LH.

METHOD OF STUDY: We have taken an approach involving the mutation of βhCG to eliminate cross‐reactive epitopes without affecting the natural folding of the polypeptide chain and thus the unique βhCG‐specific epitopes.

RESULTS: Several mutants have been constructed that have maintained the binding to hCG‐specific monoclonal antibodies (mAbs) but have lost the ability to bind to a panel of LH cross‐reactive mAbs. To investigate the immunogenicity of selected mutants, mice were immunized with expression plasmid DNA, containing the gene for wild‐type βhCG and two mutants: mutant 3, with four amino acid substitutions (68 Arg→Glu; 74 Arg→Ser; 75 Gly→His; 79 Val→His), and mutant 7, with a single amino acid substitution (68 Arg→Glu).

CONCLUSIONS: Athough both mutants were able to elicit antibody responses in at least some animals, the levels were less than those seen with the wild‐type βhCG DNA, and there seems still to be a residual cross‐reacitivity with LH. Attempts to improve the immunogenicity of the mutants and to further modify the sequence to remove the cross‐reacitivty are currently underway.
Antisperm Antibodies in Prepubertal Boys and Their Reactivity with Antigenic Determinants on Differentiated Spermatozoa
Authors: DOMAGAŁA A. , KAMIENICZNA M. , KOWALCZYK D. , KURPISZ M.
Published: September 2011

PROBLEM: Antisperm antibodies induced in prepubertal boys with testicular failures were characterized by using four techniques of antibody detection. The reactivity of circulating antisperm antibodies in prepubertal boys and the reactivity of antibodies in sera samples of adult fertile and infertile males were compared against the same sperm antigenic pools (live or fixed spermatozoa, or sperm antigenic extracts).

METHOD OF STUDY: The incidence of antisperm antibodies in sera samples of 69 prepubertal boys with testicular failures and 21 samples obtained from adult, male individuals was assessed by indirect immunobead binding test (IDIBT), flow cytometry measurement, enzyme‐linked immunosorbent assay, and Western blotting. Immunoblot analysis was performed by using sperm extracts of glycosylated and deglycosylated solubilized membrane antigens.

RESULTS: Sera samples were studied in a group composed of healthy prepubertal boys (n = 7) and prepubertal boys with testicular failures (n = 69). Applied tests of antibody detection revealed striking differences in a group of boys with testicular pathology. With IDIBT, 7% of the sera samples were found positive, whereas with flow cytometry measurement, 48% of the sera samples were positive. Immunosorbent assay (fixed sperm) indicated 32% positive cases in the same group. The sera samples were found to be positive in 65% of immunoblotting reactions with glycosylated antigens and in 70% of immunoblotting reactions with deglycosylated antigens. All applied detection assays were clearly negative on sera samples from fertile, adult males. Western immunoblotting indicated an immunodominant antigenic determinant of 58 kDa.

CONCLUSIONS: Tests of antibody detection with the use of live sperm (IDIBT and flow cytometry measurements) presented low sensitivity (8% and 48%, respectively) in a group of prepubertal boys. This observation underlines the difficulties in assigning the prospective prognosis of future fertility status in prepubertal boys with antisperm antibodies.
Expression and Localization of the Th2‐type Cytokine Interleukin‐13 and Its Receptor in the Placenta During Human Pregnancy
Authors: DEALTRY GILLIAN B. , CLARK DAWN E. , SHARKEY ANDREW , CHARNOCK‐JONES D. STEPHEN , SMITH STEPHEN K.
Published: September 2011

PROBLEM: To investigate the expression of the Th2‐type cytokine interleukin (IL)‐13 and its receptor in human placenta during gestation.

METHOD OF STUDY: Expression of IL‐13 and its receptor was analyzed by reverse transcriptase (RT)‐polymerase chain reaction (PCR), in situ hybridization and immunohistochemistry using human placental samples.

RESULTS: IL‐13 mRNA was detected by RT‐PCR in placental extracts from all stages of gestation. In situ hybridization revealed IL‐13 mRNA in first trimester cytotrophoblast and syncytiotrophoblast. Few positive cells were found within decidual sections from the same pregnancy. Immunohistochemistry revealed a similar pattern to in situ hybridization. This was largely absent in second‐ and third‐trimester placentae. IL‐13 receptor α chain (IL‐13Rα) was detected by immunofluorescence on the surface of leukocytes in first‐trimester villous core and decidua.

CONCLUSIONS: The expression of IL‐13 is spatially and temporally regulated in the placenta, and associated with the close proximity of a receptor‐bearing target leukocyte population.
Expression of a Progelatinase Activator (MT1‐MMP) in Human Fetal Membranes
Authors: Fortunato Stephen J. , Menon Ramkumar , Lombardi Salvatore J.
Published: September 2011

PROBLEM: The finding of MMP‐2 (which degrades type IV collagen) and TIMP‐2 (the tissue inhibitor of MMP) in fetal membranes suggests the possibility of membrane self‐destruction as an etiology of premature rupture of fetal membranes. MMP‐2 is activated by a membrane‐bound MMP (MT1‐MMP). This study was undertaken to detect the presence of MT1‐MMP in human fetal membranes.

METHOD OF STUDY: Fetal membranes were placed in an organ explant system and stimulated with lipopolysaccaride (LPS). MT1‐MMP expression was studied in frozen tissues by reverse transcriptase (RT)‐polymerase chain reaction (PCR) using primers designed in our laboratory. DNA sequence analysis was performed to verify the specificity of PCR products. In situ hybridization and immunocytochemistry were used to localize MT1‐MMP mRNA and peptide, respectively.

RESULTS: RT‐PCR data indicated the presence of mRNA for MT1‐MMP in fetal membranes. Although PCR is not quantitative, no differences in mRNA band intensities were noticed after LPS stimulation. MT1‐MMP expression was constitutive throughout the culture period. In situ hybridization demonstrated amnion, chorionic laeve, cytotrophoblast cells, and the cells in the reticular and spongy layer of the extracellular matrix as the origin of MT1‐MMP mRNA and peptide.

CONCLUSIONS: This is the first study documenting the amniochorionic membrane as a source of MT1‐MMP mRNA and peptide. Activation of progelatinase A requires the presence of this membrane‐associated MMP. The finding of MT1 ‐MMP in a tissue already known to produce MMP‐2 and TIMP‐2 documents the full system for activation and inhibition of this gelatinase. During infection, an imbalance in the expression of MT1‐MMP, MMP‐2, and TIMP‐2 may constitute an endogenous pathway of membrane degradation.
Histologie Distribution and Biochemical Properties of α1‐Microglobulin in Human Placenta
Authors: Berggård Tord , Enghild Jan J. , Badve Sunil , Salafia Carolyn M. , Lögdberg Lennart , ÅKerströum Bo
Published: September 2011

PROBLEM: The embryo is protected from immunologic rejection by the mother, possibly accomplished by immunosuppressive molecules located in the placenta. We investigated the distribution and biochemical properties in placenta of the immunosuppressive plasma protein α1‐microglobulin.

METHOD OF STUDY: Placental a α1‐microglobulin was investigated by immunohistochemistry and, after extraction, by electrophoresis, immunoblotting and radioimmunoassay.

RESULTS: α1‐Microglobulin staining was observed in the intervillous fibrin and in syncytiotrophoblasts, especially at sites with syncytial injury. Strongly stained single cells in the intervillous spaces and variably stained intravillous histiocytes were noted. Solubilization of the placenta‐matrix fraction and placenta membrane fraction released predominantly the free form of α1‐microglobulin, but, additionally, an apparently truncated form from the placenta‐membrane fraction. The soluble fraction of placenta contained two novel α1‐microglobulin complexes.

CONCLUSIONS: The biochemical analysis indicates the presence in placenta of α1‐microglobulin forms not found in blood. The histochemical analysis supports the possibility that α1‐microglobulin may function as a local immunoregulator in the placenta.
Immunopathology of the Implantation Site Utilizing Monoclonal Antibodies to Natural Killer Cells in Women with Recurrent Pregnancy Losses
Authors: Kwak Joanne Y.H. , Beer Alan E. , Kim Sung Hee , Mantouvalos Harris P.
Published: September 2011

PROBLEM: Placental lesions of 71 women with documented recurrent spontaneous abortions of unknown etiology were evaluated using immunohistochemical staining.

METHOD OF STUDY: Placental tissue blocks (less than 12 weeks gestation) from prior pregnancy losses were obtained, recut, and analyzed utilizing monoclonal antibody to identify the trophoblast (cytokeratin 8/18) and natural killer (NK) cells (CD57) at the implantation site. The following features were evaluated: trophoblast invasion pattern; syncytium formation; vasculitis and thromboembolism of decidual vessels: decidual inflammation; decidual necrosis; fibrin deposition at the decidual necrosis site; mononuclear‐cell infiltration in villi and intervillous space; perivillous fibrin deposition; trophoblast morphology; and quantitation of CD57 + NK cells within the decidual tissue near the implantation site. Controls consisted of 20 healthy women with no history of recurrent pregnancy losses, who had their pregnancies electively terminated.

RESULTS: Of the women studied, 29.6% demonstrated elevated CD57 + NK cells at the implantation site (P = 0.030), 54.1% had inadequate cytotrophoblast invasion depth (P = 0.000), 44.1% demonstrated inadequate syncytium formation (P = 0.004), and 33.9% presented thromboembolism in decidual vessels (P = 0.025).

CONCLUSION: Some women with recurrrent spontaneous abortions demonstrate abnormal placental lesions at the implantation site. Immunopathologic evaluation of the placental implantation site that terminated in a spontaneous abortion may reveal the immunopathogenesis of previous pregnancy losses.
The Expression of Human Leukocyte Antigen‐G on Trophoblasts Abolishes the Growth‐Suppressing Effect of Interleukin‐2 towards Them
Authors: Hamai Yoko , Fujii Tomoyuki , Yamashita Takahiro , Miki Akinori , Hyodo Hironobu , Kozuma Shiro , Geraghty Daniel E. , Taketani Yuji
Published: September 2011

PROBLEM: We have shown the attenuated human leukocyte antigen (HLA)‐G expression on trophoblasts and an aberrant expression of interleukin (IL)‐2, a cytotoxic cytokine, in decidual tissue in preeclampsia, where deteriorated trophoblastic invasion into decidual layers may constitute a crucial pathogenesis. We hypothesized that the absence of HLA‐G might make trophoblasts susceptible to compromise by IL‐2.

METHOD OF STUDY: We analyzed the growth of HLA‐G‐negative and ‐positive cell lines, all of which possessed IL‐2 receptors, in the culture with or without IL‐2 supplementation.

RESULTS: The proliferation of HLA‐G‐positive trophoblastic cell lines (BeWo and JEG‐3) was not influenced by the addition of IL‐2, whereas a HLA‐G‐negative trophoblastic cell line (JAR) exhibited significantly decreased proliferation when cultured with IL‐2. Interestingly, the transfection of JAR cells with HLA‐G completely eliminates the growth‐inhibitory effect of IL‐2.

CONCLUSION: The expression of HLA‐G may commit trophoblasts to evade cell damage by IL‐2, which may be relevant to maternal tolerance of the fetus during pregnancy and its derangement as exemplified by preeclampsia.
Mitogen‐Induced Cytokine Responses of Maternal Peripheral Blood Lymphocytes Indicate a Differential Th‐Type Bias in Normal Pregnancy and Pregnancy Failure
Authors: Makhseed M. , Raghupathy R. , Azizieh F. , Al‐Azemi M. M. K. , Hassan N. A. , Bandar A.
Published: September 2011

Makhseed M, Raghupathy R, Azizieh F, Al‐Azemi MMK, Hassan NA, Bandar A. Mitogen‐induced cytokine responses of maternal peripheral blood lymphocytes indicate a differential Th‐type bias in normal pregnancy and pregnancy failure. AJRI 1999; 42:273–281 © Munksgaard, Copenhagen

PROBLEM: Profiles of Th1‐ and Th2‐type cytokines were studied in women with a history of successful pregnancy and in women with a history of unexplained recurrent spontaneous abortions (RSA) with the objective of elucidating Th1‐ and Th2‐type bias in normal pregnancy and pregnancy failure.

METHOD OF STUDY: Peripheral blood mononuclear cells (PBMCs) from 54 women with a history of normal pregnancy and 23 women with a history of unexplained RSA, obtained at delivery or on the day of abortion, respectively, were stimulated with phytohemagglutinin (PHA), followed by the estimation of four Th2 cytokines and four Th1 cytokines.

RESULTS: Significantly greater levels of Th2 cytokines were produced by the normal group than by the RSA group. On the other hand, significantly higher levels of Th1 cytokines were produced by the RSA group than by the normal pregnancy group.

CONCLUSIONS: These data support the concept that unexplained recurrent spontaneous abortion is associated with an increase in Th1‐type reactivity, while Th2 dominance is a feature of successful pregnancy.
An Inverse Relation Between the Expression of Tumor Necrosis Factor Alpha (TNF‐α) and TNF‐α Receptor in Human Endometrium
Authors: Chegini Nasser , Dou Quigchuan , Williams R. Stan
Published: September 2011

Chegini N, Dou Q, Williams RS. An inverse relationship between the expression of tumor necrosis factor alpha (TNF‐α) and TNF‐α receptor in human endometrium. AJRI 1999; 42:297–302 © Munksgaard, Copenhagen

PROBLEM: To determine whether the expression of tumor necrosis factor (TNF‐α) correlates with TNF‐α receptor expression in human endometrium.

METHOD OF STUDY: A multiprimer synthetic cDNA standard template containing complimentary sequences for several cytokines including TNF‐α and TNF‐α receptor type 2 was constructed and used in quantitative reverse transcription polymerase chain reaction (Q‐RT‐PCR).

RESULTS: Endometrium from proliferative phase of the menstrual cycle expresses higher levels of TNF‐α mRNA (2.35 ± 0.2 × 105 copies/μg of total cellular RNA) than secretory phase ([1.3 ± 0.08 × 105 copies] P < 0.05), with a significant reduction during menses (1.2 ± 0.1 × 104 copies) and postmenopausal period (8.1 ± 1.6 × 104 copies [P < 0.05]). In contrast, TNF‐α type 1 receptor mRNA expression was higher in endometrium from the secretory phase (6.6 ± 0.6 × 107 copies) compared to the menses (5.1 ± 0.5 × 106 copies), proliferative phase (1.9 ± 0.1 × 106 copies) and postmenopausal period (5.8 ± 0.7 × 104 copies [P < 0.05]). Comparatively, TNF‐α receptor type 2 is expressed 10 to 100 fold higher in the endometrium than TNF‐α (P < 0.05).

CONCLUSION: The data confirm that human endometrium expresses TNF‐α; and provide the first evidence that TNF‐α expression is inversely related to TNF‐α type 1 receptor expression during the menstrual cycle. Such an inverse relation between TNF‐α and TNF‐α receptor expression may provide a regulatory mechanism necessary to overcome the detrimental effect of high levels of TNF‐α on various endometrial cell types.