bluuebunny/crossref_metadata_2025 huggingface dataset:
Platelet Thromboxane A2 Receptor Number and Function in Normal and Hypertensive Pregnancy
Authors: DI RENZO GIAN CARLO , COATA GIULIANA , DE FRANCESCHI SERENA , CHIARADIA ELISABETTA , KETT ATTILA , VEZZA ROBERTA , NENCI GIUSEPPE G. , COSMI ERMELANDO V. , GRESELE PAOLO
Published: May 2013

PROBLEM: Our study evaluated the number and function of platelets for thromboxane A2/prostaglandin H2 (TxA2/PGH2) and the platelet response to TxA2 receptor agonists in normal and hypertensive pregnancy. In addition, correlations between platelet membrane lipid composition and TxA2 receptor number and function were evaluated.

METHODS: Ten normotensive healthy pregnant women (GC), 10 hypertensive pregnant women (PIH), and 10 normotensive nonpregnant healthy women (C) were examined. Radioligand binding and aggregation studies were performed. Lipid composition of platelet membrane was assessed as cholesterol to phospholipids ratio (CH/PL) and normalized to protein content.

RESULTS: TxA2/PGH2 receptor binding sites and affinity did not differ among the groups. An enhanced response to TxA2 receptor agonist U 46619 in GC and in PIH compared to C was observed, while platelet response to thrombin was not different among the groups. CH/PL was altered in PIH respect to GC and C. No significant correlation was found between CH/PL and number and function of platelet TxA2 receptors in PIH.

CONCLUSIONS: In PIH there is no alteration of number or function of TxA2 receptors on platelet membrane. However, the enhanced TxA2 production associated with the increased sensitivity of platelet to TxA2 in pregnancy may contribute to the microcirculatory thrombotic changes of PIH.
Detection of Fetal Trisomies 21 and 18 From Maternal Blood Using Triple Gradient and Magnetic Cell Sorting
Authors: GÄNSHIRT‐AHLERT DOROTHEE , BÖRJESSON‐STOLL REGINA , BURSCHYK MONIKA , DOHR ANGELIKA , GARRITSEN HENK S.P. , HELMER ELISABETH , MINY PETER , VELASCO MARTA , WALDE CORINNA , PATTERSON DAVID , TENG NELSON , BHAT NEELIMA M. , BIEBER MARCIA M. , HOLZGREVE WOLFGANG
Published: May 2013

PROBLEM: The need for an inexpensive and reproducible technique for noninvasive prenatal diagnosis by fetal cell isolation from maternal blood.

METHOD: For enrichment of fetal cells we used a combination of triple density gradient and magnetic sorting (MACS) of (anti‐CD71) transferrin receptor antibody labeled cells followed by fluorescence in situ hybridization (FISH) with chromosome‐specific DNA probes for detection of fetal aneuploidies. We identified 15 cases of fetal trisomy (five cases with a trisomy 18 and ten cases with a trisomy 21) with subsequent chromosome‐specific FISH.

RESULTS: We found in all of the aneuploid pregnancies that the percentage of cells with three hybridization signals did not overlap with those of normal controls independent from gestational ages and previous invasive procedures.

CONCLUSIONS: Our new approach for noninvasive prenatal diagnosis has proven to be reliable in this first series.
Serum Soluble Interleukin‐2 Receptor α Levels in Patients With Gynecologic Cancers: Early Effect of Surgery
Authors: BARTON DESMOND P.J. , BLANCHARD D. KAY , MICHELINI‐NORRIS BEATRIZ , ROBERTS WILLIAM S. , HOFFMAN MITCHEL S. , FIORICA JAMES V. , NICOSIA SANTO V. , CAVANAGH DENIS , DJEU JULIE Y.
Published: May 2013

PROBLEM: To determine the early effect of abdominal cytoreductive surgery on serum soluble interleukin‐2 receptor α (sIL‐2Rα) levels in patients with ovarian and cervical cancer, and to determine if the extent of cytoreduction correlated with the changes in serum sIL2Rα.

METHOD: Thirty‐nine patients with gynecologic cancer had serum sIL‐2Rα measured by enzyme‐linked immunosorbent assay before abdominal cytoreductive surgery and once in the early postoperative period.

RESULTS: Only patients with advanced epithelial ovarian cancer had elevated preoperative serum sIL‐2Rα levels. In 20 of 25 ovarian cancer cases (80%) and 10 of 14 cervical cancer cases (71.4%) the postoperative serum sIL‐2Rα levels exceeded the preoperative level (P = .003 and P = .011, respectively). Overall, the mean postoperative serum sIL‐2Rα level was greater than the preoperative level (P = .0001).

CONCLUSION: Patients with early stage gynecologic cancer did not have elevated serum sIL‐2Rα levels before surgery. In the early postoperative period the serum sIL‐2Rα level was increased, which may be a nonspecific response to the trauma of surgery. Soluble IL‐2Rα may be one of the factors responsible for the immunosuppression in the early postoperative period, but may also herald a surge of activated T cells.
Characterization of an Immunoregulatory Cell Subset Involved in the Upregulation of the Autoimmune Response to Rat Male Accessory Glands
Authors: IRIBARREN PABLO , CORREA SILVIA G. , RIERA CLELIA M.
Published: May 2013

PROBLEM: We studied the regulation of the autoimmune response to rat male accessory glands (RAG) using liposomes as adjuvants.

METHOD: Adult male Wistar rats were submitted to three intraperitoneal (i.p.) immunizations with 750 μg of saline extract of RAG associated with liposomes. The delayed type hypersensitivity (DTH) response studied approximately 10 days after each immunization developed after the first immunization, having a remission state after the second one and a clear increase after the third injection. In a further study, spleen mononuclear (SpM) cells obtained form immunized rats 10 days after the third immunization (DTH positive) or from normal rats were separated as adherent (VV +) or nonadherent (VV —) to Vicia villosa population. In VV + SpM cells from immunized or normal animals an enhanced percentage of OX8 + cells (P<.05 and P< .01, respectively) was found, but in VV — SpM cells from the same groups of rats an enhanced percentage of W3/25 + cells (P <.01 and P <.05, respectively) was found when they were studied by immunofluorescence. Later on, we transferred total VV + or VV — SpM cells from i.p. immunized rats to immunized recipients 10 days after the second immunization (DTH negative). The DTH response was enhanced in recipients of total or VV+ SpM cells (P<.01). It was also observed that the transfer of VV — SpM cells from immunized rats or total or VV + SpM cells from normal rats did not reduce the suppression state observed after the second injection (P = NS). The total SpM cells obtained 10 days after the third immunization (DTH positive) were able to transfer autoimmune response to RAG to normal animals (P<.01), whereas VV + SpM cells did not show that capacity (P = NS).
T‐Cell Receptors Are Expressed but Down‐Regulated on Intradecidual T Lymphocytes
Authors: MORII TAKESHI , NISHIKAWA KIYOSHI , SAITO SHIGERU , ENOMOTO MASAHIRO , ITO AYAKO , KURAI NOBUO , SHIMOYAMA TAKETO , ICHIJO MOTOHIKO , NARITA NOBUHIRO
Published: May 2013

PROBLEM: Dietl et al. considered “intradecidual T cell tolerance towards fetal antigens” with their observation that intradecidual T cells lack immunohistochemically detectable amounts of T cell receptor (TCR) molecules while expressing normal amounts of CD3 molecules during early normal pregnancy.

METHOD: To reevaluate these findings we examined the TCR and CD3 expression on intradecidual T cells using flow cytometry.

CONCLUSIONS: In our study, all intradecidual CD3+ T cells expressed either TCR αβ or TCR γδ. However, the expression of the CD3/TCR complex on intradecidual T cells was down‐regulated. The level of CD3/TCR complex expression on intradecidual T cells was about two‐thirds of that on peripheralblood T cells. Further, the proportions of αβ + and γδ + cells in CD3 + cells did not significantly differ between decidua and peripheral blood.
Phenotypic Characterization of CD7 +, CD3 +, and CD8+ Lymphocytes From First Trimester Human Decidua Using Two‐Color Flow Cytometry
Authors: CHERNYSHOV VICTOR P. , SLUKVIN IGOR I. , BONDARENKO GENADY I.
Published: May 2013

PROBLEM: There is increasing evidence that decidual lymphocytes play a major role in local interactions at the fetomaternal interface.

METHOD: In this paper we use two‐color flow cytometry to delineate the phenotype of lymphocytes obtained from human early decidua by mechanical dispersal technique.

RESULTS: The most abundant decidual lymphocytes expressed CD7, CD38, CD56, and CD2 markers, relatively small proportions of CD3 +, CD8 +, CD4 +, CD16 +, CD45RA+, CD11b+, and Leu8+ cells were also present. The vast majority of decidual CD7+ lymphocytes expressed CD38, CD2, and CD56 markers and were CD3 −, CD8 −, CD 16 −, and CD57 −. Decidual CD3+ lymphocytes were weakly staining for TCR α/β, lacked T‐cell receptor (TCR) γ/δ molecules, and ∼40% of them expressed HLA‐DR. All decidual CD8+ lymphocytes were CD2+ and the majority of them expressed CD38, CD56, and CD7 markers and were CD3−at the same time CD8 + CD7− lymphocytes were found in decidua. According to the expression of the CD45RA marker, decidual CD8 + lymphocytes could be divided into two subsets: CD8 + CD45RA + CD56 + and CD8 + CD45RA −.

CONCLUSIONS: These data clearly demonstrate that decidual lymphocytes display phenotypical features different from those of their counterparts in peripheral blood.
Flow Cytometry Analysis of Lymphocyte Subpopulations in Peritoneal Fluid of Women With Endometriosis
Authors: Oosterlynck Didier J. , Meuleman Christel , Lacquet Françoise A. , Waer Mark , Koninckx Philippe R.
Published: May 2013

PROBLEM: We investigated the lymphocyte subpopulations in peripheral blood (PB) and peritoneal fluid (PF) of women with and without endometriosis to evaluate if the decreased natural killer (NK)‐mediated cytotoxicity in women with endometriosis was due to a quantitative defect or not.

METHOD: The PB and PF mononuclear cells of 59 women undergoing a diagnostic laparoscopy for pain and/or infertility were analyzed by flow cytometry.

RESULTS: The number and concentration of PF mononuclear cells (MC) was increased in women with endometriosis compared to women without endometriosis. The monocyte/macrophage marker (CD14) was expressed on 70.3 and 66.9% of PFMC of women with and without endometriosis, respectively. The CD4/CD8 ratio was inverted in the PF, and this was more pronounced in women with endometriosis. In the PF of women with endometriosis, 41.3% of the lymphocytes were CD8 positive, compared to 34.3% in women without endometriosis. The percentage of NK positive lymphocytes in PF, using three different monoclonal antibodies directed against NK cell markers (CD57, CD 16, and CD56) were not different between women with and without endometriosis. In women with endometriosis, 12.7,9.5, and 28.8% of lymphocytes were CD57, CD16, and CD56 positive, respectively.

CONCLUSION: PFMC consisted mainly of phagocytic and human leukocyte antigen (HLA)‐restricted or HLA unrestricted cytotoxic cells capable of reacting to various antigens entering the cavity from the lower genital tractus. Furthermore, the decreased NK activity reported in PB and PF of women with endometriosis was not likely to be caused by a quantitative defect, since the percentage of NK positive lymphocytes was not different between women with and without endometriosis.
Secretory Immunity in the Female Reproductive Tract
Authors: Kutteh William H. , Mestecky Jiri
Published: May 2013

PROBLEM: Antibodies and antibody‐producing cells display a different and characteristic distribution in body fluids and tissues.

METHOD: We have investigated the tissues of the female reproductive tract to determine whether the distribution of immunoglobulin‐producing cells and the contents of cervical secretions were similar to those found in tissues of the secretory immune system.

RESULTS: Immunohistochemical examinations of female genital tissues revealed the presence of plasma cells that secrete IgA (and in lower numbers IgM and IgG) especially in the subepithelial layers of the uterine endo‐ and ectocervix, fallopian tubes, and vagina. Both IgA1‐ and IgA2‐producing plasma cells were found in approximately equal proportions. The presence of J‐chain in the IgA‐secreting cells suggests the synthesis of polymeric IgA (plgA). Epithelial cells lining the fallopian tube and endocervix were positive for secretory component (SC), which is required for the transepithelial transport of pIgA into external secretions. Cervical mucus was collected and the molecular forms of IgA were separated using column chromatography. Approximately 80% of IgA in cervical mucus was polymeric compared with 55% in the vaginal fluid.

CONCLUSIONS: These data indicate that all effector components of the mucosal immune system are present in the female reproductive tract. The immunization routes that lead to a secretory IgA (S‐IgA) response need to be further explored.
Age, Environment, and Lymphocyte Immunization Influence the Spontaneous Resorption Rate in the CBA/J × DBA/2J Mouse Model
Authors: Ho Hong‐Nerng , Chen Shee‐Uan , Yang Yu‐Shih , Huang Su‐Cheng , Lee Tzu‐Yao , Gill Thomas J.
Published: May 2013

PROBLEM: This study was designed to examine the influence of age, environment, and lymphocyte immunotherapy on fetal resorption rates in the CBA/J × DJA/2J mouse model.

METHODS: After weaning, CBA/J female mice were randomly allocated to two different environments: room A was a conventional housing facility, and room B was a specific pathogen‐free room. They were further divided into two groups in each room according to age (> three months and ≤ three months), and mated with CBA/J, DBA/2J, or BALB/c males. The fetal resorption rates were observed. The immunization study was conducted using CBA/J females greater than three months old in room B. Preimmunization with various preparations and dosages of lymphoctyes was performed 1 wk before mating with DBA/2J males, and fetal resorption rates were measured to investigate the effect of immunotherapy.

RESULTS: In room A, the fetal resorption rates (9.4 to 11.8%) were not significantly different among the various mating combinations using CBA/J females greater or less than three months of age. In room B, CBA/J females older than three months and mated with DBA/2J males had a higher rate of fetal resorption than those younger than three months (28.9 versus 14.2%) and had higher rate of fetal resorption than those housed and mated in room A (28.9 versus 10.9%). This fetal resorption rate was also higher than those seen with other mating combinations. Preimmunization with male BALB/c splenic lymphocytes (optimal dose 1 × 107 cells) was effective in decreasing fetal resorption rate (7.9%), whereas administration of normal saline, or immunization with male CBA/J or DBA/2J lymphocytes was not (20.7 to 27.0%).

CONCLUSION: Age and environment influence the spontaneous resorption rate in the CBA/J × DBA/2J mouse model. The high fetal resorption rate of older CBA/J females mated with DBA/2J males in a pathogen‐free environment can be reduced by immunization with BALB/c splenic lymphocytes. These results suggested that a variety of mechanisms might initiate early pregnancy failure and that immunological modulation during implantation might be a nonspecific factor.
Incidence of Autoimmune Antibodies in Failed Embryo Transfer Cycles
Authors: Birkenfeld A. , Mukaida T. , Minichiello L. , Jackson M. , Kase N.G. , Yemini M.
Published: May 2013

PROBLEM: The presence of antiphospholipid antibodies lupus anticoagulant (LAC), anticardiolipin antibody (ACA) as well as antinuclear antibody (ANA) has been associated with early spontaneous pregnancy loss and adverse pregnancy outcome. The purpose of this study was to investigate the possible role of autoimmune antibodies (LAC, ACA, and ANA) as a cause of implantation failure following embryo transfer (ET) after in vitro fertilization (IVF).

METHOD: Three groups were studied: Group I, 56 patients who failed to conceive following ET; group II, 14 patients who have conceived following IVF‐ET and delivered or are carrying an uncomplicated ongoing pregnancy; and group III, 69 patients who were new candidates for IVF‐ET.

RESULTS: Eighteen out of 56 (32.1%) of patients who failed to conceive following previous IVF‐ET cycle (group I) tested positive for one or more of the autoimmune antibodies. None of the 14 patients of group II tested positive for autoimmune antibodies (P<.02). Seven out of the 69 patients (10%) of group III were found positive to one or more of the autoimmune factor. This rate is significantly lower than the rate of positive autoimmune antibodies detected in group I (P<.003). Fifteen patients of the 18 who tested positive for autoimmune antibodies and who had previously failed to conceive following ET underwent a subsequent IVF‐ET cycle while being treated with prednisone and aspirin. Seven out of the 15 (46.6%) conceived and were able to sustain a clinical ongoing pregnancy.

CONCLUSIONS: Patients receiving ET are carrying viable embryos within the intrauterine environment. Therefore, in this unique group of patients, failure to demonstrate a positive pregnancy test represents an implantation failure or a very early postimplantation loss. The results of this study suggest that periimplantation events may be affected by autoimmune antibodies. Very early miscarriage or implantation failure may be related to the same pathophysiological mechanism that causes recurrent miscarriages and is diagnosed incorrectly as infertility.
Effect of Epidermal Growth Factor on Mouse Sperm Acrosome Reaction Induced by Zona Pellucida
Authors: Furuya Satoru , Endo Yoshihiro , Oba Mikiko , Matsui Yukari , Suzuki Shuetu , Nozawa Shiro
Published: May 2013

PROBLEM: The effect of epidermal growth factor (EGF) on the acid‐solubilized zona pellucida (ZP)‐induced acrosome reaction was investigated in mouse sperm.

METHOD: Mouse epididymal sperm were capacitated in modified Krebs‐Ringer bicarbonate buffer (m‐KRB) for 120 min and further treated with acid‐solubilized ZP(4 zona/μl) for an additional 60 min to induce the acrosome reaction. The chlortetracycline fluorescence assay was used to monitor the acrosome reaction. The acrosome reacted sperm featured the acrosome reacted (AR) pattern, which demonstrates the lack of fluorescence on the head and bright midpiece.

RESULTS: EGF caused an early increase in the AR pattern in response to the acid‐solubilized ZP in a dose‐dependent manner. The EGF‐dependent stimulation of the ZP‐induced acrosome reaction was inhibited by an inhibitor of protein tyrosine kinases, genistein or activators of Ca++ and phospholipid‐dependent protein kinase (protein kinase C). Furthermore, the stimulatory effect of EGF was not attenuated when sperm were capacitated in the presence of islet‐activating protein, an inactivator of inhibitory guanine nucleotide‐binding regulatory protein (Gi protein).

CONCLUSION: EGF stimulates the ZP‐induced acrosome reaction in a manner that is independent of the Gi protein. The EGF action is regulated by protein tyrosine kinase and protein kinase C.
Effects of Atrial Natriuretic Peptide on Rat Ovarian Granulosa Cell Steroidogenesis in Vitro
Authors: Johnson K.M. , Hughes F.M. , Fong Y.‐Y. , Mathur R.S. , Williamson H.O. , Gorospe W.C.
Published: May 2013

PROBLEM: The localization and biological effects of atrial natriuretic peptide (ANP) are not limited to cardiac tissue but extend to a number of extra‐atrial tissues and organs, including the ovary. The objective of the present study was to determine the effects of ANP on granulosa cell steroidogenesis. Hence, the direct effects of ANP on the production of progesterone, 20α‐hydroxpregn‐4‐en‐3‐one (20α‐OH‐P), and estrogen by undifferentiated and differentiated rat ovarian granulosa cells were examined in vitro.

METHOD: Undifferentiated granulosa cells obtained from the ovaries of diethylstilbestrol‐primed, immature rats were treated with increasing doses (10−12 to 10−6 M) of rANP for 48 h.

RESULTS: ANP evoked a two‐ and threefold increase in progesterone and 20α‐OH‐P production relative to untreated controls, respectively. Increasing doses of ANP in combination with porcine FSH (125 ng/well) resulted in a biphasic response in progesterone production above FSH‐treated controls. Specifically, a maximal inhibition of 35% in progesterone production was achieved at 10−9 M ANP, followed by a stimulation to levels comparable with FSH‐stimulated controls at higher doses examined. Increasing doses of ANP evoked a twofold increase in FSH‐stimulated 20α‐OH‐P production over respective controls. Following pretreatment of granulosa cells with FSH for 48 h to evoke differentiation, ANP caused a significant dose‐dependent inhibition in basal progesterone production that resulted in a 76% suppression at the highest dose examined. In contrast, ANP evoked a 2.8‐fold increase in 20α‐OH‐P production when compared with controls. Finally, in FSH‐stimulated differentiated granulosa cells, ANP evoked a threefold increase in progesterone production and a 65% inhibition in 20α‐OH‐P production. ANP exerted no significant effects on estrogen production by either undifferentiated or differentiated granulosa cells in the presence or absence of FSH.

CONCLUSIONS: These data demonstrate that ANP can modulate directly progestin steroidogenesis in both undifferentiated and differentiated rat ovarian granulosa cells in vitro and, therefore, may play an important role in granulosa cell differentiation and follicular maturation.
Functions of Macrophages in Human Decidual Tissue in Early Pregnancy
Authors: Mizuno Masahiko , Aoki Koji , Kimbara Toshihiro
Published: May 2013

PROBLEM: Roles of decidual macrophages (DMs) in the maintenance of early pregnancy was determined by comparing three of their functions, antigen presentation, immunoregulation, and lymphokine production, with those of peripheral monocytes (PMos) isolated from the same subjects.

METHOD: The antigen‐presenting capacity of DMs was examined by the one‐way mixed lymphocyte reaction (MLR) in which accessory cell‐depleted mononuclear cells isolated from pregnant women were used as responders. The effect of DMs on cellular immunity was investigated by inhibition tests of either one‐way MLR or phytohemagglutinin stimulation. The production of interleukin‐1 (IL‐1) and prostaglandin E2 (PGE2) by DMs following lipopolysaccharide stimulation was examined.

RESULTS: Addition of increasing concentrations of DMs to the culture resulted in a concentration‐dependent proliferative response, as well as with PMos. In both assays, a stronger suppression was observed in the presence of DMs from normal pregnant women compared with PMos from the same subjects. DMs were found to secrete significantly lower levels of IL‐1 α and IL‐1β than PMos. No difference in PGE2 production was observed between DMs and PMos.

CONCLUSIONS: These findings suggest that DMs present in human early decidual tissue have a capacity for allo‐antigen presentation, a higher suppressive activity, and a lower capacity to produce IL‐1 than their peripheral counterparts.
Interleukin‐1 Receptor Antagonist (IL‐1ra) Production by Human Amnion, Chorion, and Decidua
Authors: Fidel P.L. , Romero R. , Ramirez M. , Cutright J. , Edwin S.S. , Lamarche S. , Cotton D.B. , Mitchell M.D.
Published: May 2013

PROBLEM: This study was conducted to determine whether (1) conditioned media from unstimulated primary cultures of human amnion, chorion, or decidua contain detectable concentrations of IL‐lra in vitro, and (2) bacterial endotoxin (LPS), tumor necrosis factor‐alpha (TNF‐α), or IL‐1‐beta (IL‐1β) stimulate amnion, chorion, or decidua to produce increased amounts of IL‐1ra.

METHOD: Placentae were obtained from women at term with intact membranes before the onset of labor. Amnion, chorion, and decidual cells were isolated by standard procedures and grown to confluence. Cells were then cultured in quadruplicate for 16 h in tissue culture medium supplemented with 10% fetal calf serum or, additionally, with various concentrations of Eschrichia coli LPS, TNF‐α, or IL‐1β. Culture supernatants were collected, and concentrations of IL‐1ra were quantitated by a sensitive and specific enzyme‐linked immunosorbent assay for IL‐lra.

RESULTS: Results showed that primary cultures of amnion and chorion from 4 of 9 and decidua from 10 of 12 placentae had detectable rates of production of IL‐lra (ranges: 0.08‐6.5, 0.42‐12.1, and 1.55–96.5 pg IL‐1ra/μg protein/16 h, respectively). In addition, LPS (10‐1,000 ng/ml) and IL‐1β (0.1–10 ng/ml), but not TNF‐α (0.01–100 ng/ml), stimulated decidual cells to release/secrete increased amounts of IL‐lra compared with media alone (range: 2.5–400 pg IL‐lra/μg protein/16 h, P < 0.0001). In contrast, neither LPS, TNF‐α, or IL‐1β could stimulate amnion or chorion to release/secrete IL‐1ra.

CONCLUSIONS: These results indicate (1) that amnion, chorion, and predominantly decidua, can release or secrete IL‐1ra in vitro, and (2) that LPS and IL‐1β can stimulate decidual cells to produce increased amounts of IL‐lra.
Maternal—Fetal Transport of Immunoglobulin G and Its Subclasses During the Third Trimester of Human Pregnancy
Authors: Malek Antoine , Sager Ruth , Schneider Henning
Published: May 2013

PROBLEM: We determined the maternal‐fetal transport of immunoglobulin G (IgG) during the third trimester of human pregnancy.

METHOD: The concentration of IgG and its subclasses (IgG1–4) was determined in sera of blood samples from 38 pregnancies collected at the time of delivery from a peripheral maternal vein (MV) and from the placental umbilical artery (UA) and vein (UV). Gestational age varied between 28 and 42 weeks (WG).

RESULT: Whereas placental weight showed a significant correlation with gestational age, the maternal level of IgG and the ratio of its subclasses did not vary with gestational age. At 28–33 WG (n = 15) the mean values in the UA (5.91 ± 1.53 g/1) and UV (6.41 ± 1.57 g/1) for total IgG concentration were lower than in the MV (10.74 ± 2.55 g/1). At the end of gestation (37–42 WG, n = 12), IgG in both UA (11.21 ± 1.95 g/1) and UV (12.26 ± 2.06 g/1) exceeded the maternal concentration (9.69 ± 1.84 g/1). In addition to the significant positive correlation between IgG concentration in the fetal circulation (UV + UA) and gestational age (28–42 WG), an increase in the positive difference between UV and UA at the end of pregnancy indicates that there is a substantial rise in placental IgG transport capacity.

CONCLUSION: All four subclasses IgG1–4 were detectable in the umbilical circulation (28–42 WG). Whereas IgG3 and IgG4 in UA and UV had a similar concentration as in MV and remained unchanged, IgG2 increased with gestation from 0.67/0.74 g/1 (UA/UV, 28–33 WG) to 1.29/1.58 g/1 (UA/UV, 37–42 WG), but nevertheless remained lower than the level found in the MV (2.65 ± 1.12 g/1). The main increase in IgG concentration, however, was due to the substantial rise in the transport of IgG1, which increased from 4.37 ± 1.24 (UA) and 4.94 ± 1.52 g/1 (UV) at 28–33 WG to 8.94 ± 1.66 (UA) and 10.89 ± 1.96 g/1 (UV) at the end of gestation, which was even higher than the overall IgG concentration in MV.
H5D12 Monoclonal Antibody Identifies a 24‐kD Immunosuppressor Factor Produced by Human Chorio Carcinoma Cells
Authors: Bose Ratna , Berczi Anna
Published: May 2013

PROBLEM: The purpose of this study was to identify and characterize embryo‐associated immunosuppressor factor (EASF) secreted by chorio carcinoma cells with the help of EASF‐specific monoclonal antibody H5D12 (raised against EASF purified from embryo growth media of in vitro fertilized human ova).

METHOD: Paraffin‐embedded slides of human chorio carcinoma as well as control cell lines were prepared, and immunohistochemistry was done by the avidin‐biotin‐peroxidase technique. EASF was affinity purified using H5D12‐Sepharose 4B from culture media of cell lines and analyzed for immunosuppressive activity (by Concanavalin‐A‐induced lymphocyte proliferation assay) and molecular weight identity (by metabolic‐labelling studies with 35S‐methionine followed by immunoprecipitation and SDS‐PAGE).

RESULTS: H5D12 showed intense immunostaining of BeWo chorio carcinoma cells. Bio‐synthetic labeling studies identifies this factor as a 24‐kD molecule, and EASF bioassay indicates that this factor possesses immunosuppressive activity. No such immunosuppressive activity or similar molecules were identified when control cell lines were analyzed.

CONCLUSIONS: Monoclonal antibody H5D12 recognizes a 24‐kD factor with immunosuppressive activity that is secreted by chorio carcinoma cells, which suggests that this is a unique factor and may be one of the key regulators of reproductive functions.
Use of Lymphocyte Platelet Binding Assay for Detecting a Preimplantation Factor: A Quantitative Assay
Authors: Barnea E. R. , Lahijani K. I. , Roussev R. , Barnea J. D. , Coulam C. B.
Published: May 2013

PROBLEM: To evaluate the ability of the lymphocyte/platelet binding assay to identify a preimplantation factor (PIF).

METHOD: Percentages of binding of lymphocytes by platelets in the presence of sera from 30 known pregnant and 30 nonpregnant individuals were compared using a novel lymphocyte/platelet binding assay. The assay is performed using a combination of a heat inactivated sera with donor O+ lymphocytes, activated complement and an antibody against CD2(T11, Ortho Pharmaceuticals).

RESULTS: In nonpregnant females (23.6 ± 6.5%) and males (17.7 ± 4.7%) the percentage of lymphocytes bound by platelets was significantly different from pregnant women (56.1 ± 15.9%) (P < 0.0001). Serial sampling of blood in five women undergoing IVF/ET who had normal pregnancies showed the detection of PIF by 4 days after transfer. The lymphocyte/platelet binding assay was not influenced by hCG, progesterone and estradiol. The interassay and intraassay variabilities were <3%.

CONCLUSIONS: The lymphocyte/platelet binding assay is a simple, reproducible, specific and cost efficient assay for measurement of PIF. Application of this assay will provide investigative and diagnostic tools for identifying and monitoring early pregnancy events.
Intralipid as Treatment for Recurrent Unexplained Abortion?
Authors: Clark David A.
Published: May 2013

PROBLEM: Safe, effective, and inexpensive alternatives to partner leukocyte immunotherapy are being sought. Psychotherapy may be effective but it is uncertain what constitutes effective treatment and the form of treatment tested in cohort controlled trials is expensive. IVIG also appears effective, but is expensive.

METHOD: A published double blind randomized controlled trial in which Intralipid (Kabi Vitrum, Toronto, Ontario) was used as a control versus trophoblast membrane vesicles was reviewed. A prediction made from this data was then tested using the DBA/2‐mated CBA/ J mouse model of recurrent spontaneous abortion.

RESULTS: It can be hypothesized from the human clinical trial data that Intralipid even in small doses could be an effective antiabortion treatment. The number of patients in the published study is too small for the required degree of precision. Intralipid was highly effective in preventing abortion in mice, and protection was prolonged. This may be explained by previous data in the literature showing that Intralipid affects the reticuloendothelial system of the recipient.

CONCLUSIONS: The evidence suggests that Intralipid might be an effective treatment for human recurrent miscarriages, and injection into women who may become pregnant has been found ethically acceptable at one university center. Comparison of Intralipid to partner leukocyte immunotherapy or IVIG would be worthwhile. For adequate statistical power, this would require a large, multicenter, prognostically stratified randomized controlled trial and could be accomplished via the Recurrent Miscarriage Immunotherapy Trialists Group network.
The Effectiveness of Allogeneic Leukocyte Immunization in Unexplained Primary Recurrent Spontaneous Abortion
Authors: Daya Salim , Gunby Joanne ,   The Recurrent Miscarriage Immunotherapy Trialists Group
Published: May 2013

PROBLEM: Unexplained primary recurrent spontaneous abortion (RSA) can be viewed as a partner‐specific problem for which immunization with allogeneic leukocytes is being offered as therapy. Published data from randomized controlled trials have produced conflicting results regarding treatment effectiveness. The aim of this study was to perform a subgroup analysis of the data from a recent worldwide collaborative meta‐analysis using the raw data for patients with primary RSA entered into randomized controlled trials of immunotherapy.

METHODS: Data from randomized controlled trials in eight centers were included in this analysis. Individual patients were included only if they had had three or more spontaneous abortions, no previous pregnancy beyond 20 weeks' gestation, no identifiable cause for the abortions, and no evidence of antipaternal antibodies. Meta‐analysis by centre and logistic regression analysis were performed to determine the overall effect of treatment in achieving live birth and to identify variables that affect the prognosis for a successful outcome.

RESULTS: In the meta‐analysis by center, immunotherapy significantly improved the live birth rate (common odds ratio = 1.94, 95% confidence interval (CI) = 1.20 to 3.12). In the analysis by patient, the likelihood of a successful outcome was also significantly better with treatment (relative risk = 1.46, 95% CI 1.19 to 1.69). The absolute treatment effect was 16.3% producing a number needed to treat of 6. The number of previous abortions had a significant negative correlation with live birth rate, such that for each additional pregnancy loss beyond three, the likelihood of live birth was reduced by 23%.

CONCLUSION: Allogeneic leukocyte immunization is an effective treatment for unexplained primary RSA when pretreatment antipaternal antibodies are absent. Better diagnostic tests are required to identify patients who may derive maximal benefit from this therapeutic approach.
Chymotrypsin‐Galactose Treatment of Sperm With Antisperm Antibodies Results in Improved Pregnancy Rates Following in Vitro Fertilization
Authors: KATSOFF DIANE , CHECK JEROME H. , BOLLENDORF ANIELA , BENFER KELLY
Published: May 2013

PROBLEM: To determine if chymotrypsin‐galactose (CG) treatment of sperm bound with antisperm antibodies (ASA) improves pregnancy rates (PRs) following in vitro fertilization (IVF).

METHOD: Patients with >50% ASA who failed to conceive despite six intrauterine insemination (IUI) cycles were included. Initially the sperm treatments were randomized with CG vs culture medium; subsequently only CG treatment was used.

RESULTS: There was a significantly lower fertilization rate in those patients inseminated with sperm incubated in culture medium vs CG (27% vs 47%, P<.05 t‐test). Similarly, a higher percentage of patients receiving culture medium treatment of sperm had failed fertilization (45%) compared to CG (11%). Though the clinical PRs were higher with CG (21%) than medium (9.5%), there was no statistical difference.

CONCLUSIONS: Though the percentage of sperm bound with antibodies are not reduced, we hypothesize that the CG treatment improves fertility by possibly mitigating the antagonistic action of these antibodies.
Suppression of Natural Killer Cell Activity by Monocytes Following Immunotherapy for Recurrent Spontaneous Aborters
Authors: HIGUCHI KAZUHIRO , AOKI KOJI , KIMBARA TOSHIHIRO , HOSOI NOBUYUKI , YAMAMOTO TAKASHI , OKADA HIDECHIKA
Published: May 2013

PROBLEM: Natural killer (NK) cell activity has previously been shown to decrease in normal pregnancy as compared with the nonpregnancy state. The purpose of this study was to determine NK cell activity in recurrent aborters and to investigate the kinetics of NK cell activity following immunotherapy.

METHODS: Recurrent aborters (N = 17) were immunized with husbands' mononuclear cells (1 × 108) twice during the early stage of current pregnancy. NK cell activity of recurrent aborters as well as that of normal pregnant (N = 12) and nonpregnant (N = 6) women (controls) was determined by 51Cr release assay. Monocytes were depleted from the mononuclear cell fraction and its effect on the NK cell activity was determined as well.

RESULTS: At around 5 wk of gestation, NK cell activity in recurrent aborters before treatment was significantly higher (28.0 ± 5.1%) than that in normal pregnancy (18.9 ± 4.3%) (P < 0.01). Following immunotherapy, NK cell activity of recurrent aborters (N = 13) who maintained their pregnancy decreased significantly (21.7 ± 8.9%) (P < 0.05). In contrast, NK cell activity of recurrent aborters (N = 4) who aborted their current pregnancy did not decrease. Depletion of monocytes resulted in a significant increase in NK cell activity (P < 0.05).

CONCLUSIONS: This study suggests that the immunotherapy induces suppression of NK cell activity which may contribute for the maintenance of pregnancy. Moreover, monocytes may be involved in this suppression.
Gonadotropin Releasing Hormone (GnRH) Agonist Induces Down‐Regulation of the CD3+CD25+ Lymphocyte Subpopulation in Peripheral Blood
Authors: HO G‐NERNG , CHEN HSIN‐FU , CHEN SHEE‐UAN , CHAO KUAN‐HAN , YANG YU‐SHIH , HUANG SU‐CHENG , LEE TZU‐YAO , GILL THOMAS J.
Published: May 2013

PROBLEM: To test whether GnRH agonist could alter in vivo human immune cells and whether the alteration is related to the success of pregnancy in an in vitro fertilization‐embryo transfer (IVF‐ET) program.

METHODS: Thirty‐six infertile patients were enrolled under the long protocol of GnRH agonist (buserelin acetate) and superovulation with gonadotropin from our IVF‐ET program. Peripheral B cells, NK cells, CD4+ and CD8+ T cells, and the expression of CD69, CD25, HLA‐DR, and CD71 antigens on the T cells were serially examined by dual‐color flow cytometry.

RESULTS: B cells, NK cells, CD8+ T cells, and CD71+ T lymphocyte subpopulations were not changed throughout the whole course of treatment. CD4+ T cell and CD25+ T cell sub‐populations were significantly down‐regulated when the GnRH agonist was used for approximately 2 wk. CD3+CD69+, CD3+CD25+, and CD3+DR+ lymphocyte subpopulations were increased at 7 days (during implantation) and at 14 days after embryo transfer in pregnant patients, but not in patients who failed to get pregnant.

CONCLUSIONS: The GnRH agonist had a transiently immunosuppressive effect on CD4+ and CD25+ T cells, but CD69+, CD25+, and HLA‐DR+ T cells were activated during and after successful implantation.
Anthropological Considerations Relevant to the Maturation of the Immune Response in Humans
Authors: GILL THOMAS J.
Published: May 2013

PROBLEM: The reason for the postnatal maturation of the immune response in humans was explored from the anthropological standpoint in order to gain some insight into how development in modern humans evolved and how it might have affected the immune system.

METHOD: The estimation of the length of gestation in modern humans if it had followed the same developmental pattern as in the other hominids and the higher primates was calculated by extrapolation from the lengths of gestation and the physical characteristics of these other primates compared to those of modern humans.

RESULTS: These calculations show that the gestation time of modern humans would be 3–18 months longer than the 9 months that it is if modern humans had followed a linear evolutionary development comparable to that of the other primates. The key difference between modern humans and the other primates is the rapid and extensive development of the brain that occurs in utero and in the first 12–18 months postnatally in modern humans.

CONCLUSION: The first 12–18 months postnatally in humans follows the embryonic/fetal pattern of development, whereas that of other primates does not. The maturation of the immune response in humans occurs during both the intrauterine and extrauterine phases of the embryonic/fetal pattern of development.
Prevention of Human Recombinant Intei leukin‐1β (rhIL‐1β) Embryolethality with Progesterone or Indomethacin
Authors: MARKS THOMAS A. , TRACY DANIEL E.
Published: May 2013

PROBLEM: Bropirimine and tilorone were found in earlier studies to be embryolethal when administered to Crl:TUC(SD)spf (TUC) rats on gestation day 10. Progesterone or indomethacin could, at least partially, prevent this effect. The immunomodulators appeared to mimic the luteolytic effects of PGF2α, resulting in a shutdown in progesterone release by the corpora lutea, followed by a disruption in maternal support to the pregnant uterus and embryolethality. Since bropirimine has been shown to induce interleukin‐1, and since this cytokine has been found to increase PGF2α levels in human decidual cells, the decision was made to investigate whether human interleukin‐1β might act in an analogous manner to bropirimine and tilorone.

METHOD: Bropirimine (400 mg/kg, p.o.) or rhIL‐1β (20, 30, or 40 μg/kg, s.c.) was administered on gestation day 10 to Crl:CD[BR] (CD) or TUC rats, alone and in combination with progesterone (2 mg/kg/day, s.c.) or indomethacin (0.6 mg/day, s.c, days 9–11). On gestation day 14 the dams were killed and their uterine contents examined.

RESULTS: rhIL‐1β (30–40 μg/kg) was embryolethal when administered to CD or TUC rats on gestation day 10. Progesterone or indomethacin coadministration prevented, at least partially, the embryolethality seen when rhIL‐1β was administered (30 μg/kg) to TUC rats.

CONCLUSION: Evidence was obtained in support of the hypothesis that interleukin‐1 is involved in the embryolethal actions of the immunomodulators bropirimine and tilorone.
Immunohistochemical Studies of the Adult Human Ovary: Possible Contribution of Immune and Epithelial Factors to Folliculogenesis
Authors: BuKOVSKý ANTONÍN , KEENAN JEFFREY A. , CAUDLE MICHAEL R. , WIMALASENA JAY , UPADHYAYA NIRMALA B. , VAN METER STUART E.
Published: May 2013

PROBLEM: Formation of primordial follicles in adult ovaries could be a cryptic process limited to relatively small areas of the ovarian cortex and occurring during a certain stage of the menstrual cycle. Such an event may require a specific milieu provided by factors involved in developmental processes, i.e., morphoregulatory molecules and macrophages.

METHOD: Adult human ovaries were investigated by immunohistochemistry for surface epithelium and granulosa cell markers (cytokeratin 18 and MHC class I), immune system‐related morphoregulatory molecules (Thy‐1 glycoprotein and N‐CAM), and macrophage phenotypes (CD14, CD68, and MHC class II).

RESULTS: In some ovaries 300–500 μm areas of surface epithelium were overgrown by tunica albuginea, descended into the stroma, and apparently fragmented into individual small (20–40 μm) follicle‐like cell nests. Differentiation of the surface epithelium was accompanied by macrophages and Thy‐1 glycoprotein. Small segments of surface epithelium showed N‐CAM and a lacked MHC class I expression. In such segments, clear spherical germ‐like cells migrated into the deeper stroma, associated with the microvasculature, and eventually aggregated with follicle‐like cell nests.

CONCLUSIONS: Our data suggest that surface epithelium may be involved in the formation of some primordial follicles in adult ovaries. This process, and further follicular fate, may require a precise interplay of immune system related morphoregulatory molecules and macrophages.
Detection of Antibodies Toward Epididymal Sperm Antigens—An Obligatory Step in Evaluation of Human Immunologic Infertility?
Authors: FICHOROVA R.N. , DIMITROVA E. , NAKOV L. , TZVETKOV D. , PENKOV R. , TASKOV H.
Published: May 2013

PROBLEM: To test the relative impact of epididymal versus ejaculated sperm in immunologic infertility.

METHOD: Human antibody binding to epididymal and ejaculated spermatozoa was compared by flow cytometry (FCM) since it allows quantitative analysis of viable sperm while ignoring nonsperm cells. To select sera for FCM, GAT, TAT, and ELISA were applied on 145 sera from fertile men, idiopathically infertile and varicocele patients.

RESULTS: All GAT/TAT‐positive infertile patients, a representative group of varicocele patients and the fertile control, were assessed by FCM. Higher reactivity toward epididymal sperm revealed 18/22 sera while only four out of them bound to ejaculated sperm stronger than the control. All varicocele sera were positive against epididymal while negative against ejaculated spermatozoa.

CONCLUSIONS: Epididymal sperm antigens may play a predominant role in some cases of immunologic infertility. Such patients might not be adequately diagnosed and respectively treated due to the limitations of diagnostic procedures applying only ejaculated spermatozoa.
Presence of Activated Macrophages in a Murine Model of Early Embryo Loss
Authors: DUCLOS ALAIN J. , HADDAD ELIAS K. , BAINES MALCOLM G.
Published: May 2013

PROBLEM: Even though our knowledge of the phenomenon at play at the fetoplacental interface has greatly advanced during the past years, a complete understanding of the reasons why the developing embryo is not rejected by maternal immune effector cells remains largely unknown.

METHODS: We have used immunohistochemistry with the macrophage‐specific markers F4/80 and MHC II to study the relationship between decidual infiltration and resorption in murine models of embryo loss between days 6 and 10 of gestation.

RESULTS: Analysis of day 8 CBA/J × DBA/2 pregnancies has revealed 2 distinct populations of embryos. The majority (69.4%) expressed low levels of F4/80+ cells, but a minority (30.6%) expressed much higher level of the macrophage marker. In CBA/J × BALB/c, most embryos (91.7%) expressed low numbers of F4/80+ cells. As earlier experiments established that products of activated macrophages (TNF‐α and nitric oxide) were implicated in embryo loss in this model, the activation status of the F4/80+ macrophages was assessed through the cell surface expression of MHC II. Again, a similar association was established: 30.6% of the CBA/J × DBA/2 embryos were infiltrated by significantly more MHC II+ cells than the control CBA/J × BALB/c mating. Finally, when coordinate expression of F4/80, MHC II and CD11b was assessed, it was found that an embryo significantly infiltrated by cells bearing one of the 3 markers was also heavily infiltrated by cells bearing the 2 other markers.

CONCLUSIONS: This study has shown that the augmented infiltration of the deciduum with maternal macrophages is an early event which precedes spontaneous abortion of the early embryo.
Identification of the Complement Regulatory Proteins CD46, CD55, and CD59 in Human Fallopian Tube, Endometrium, and Cervical Mucosa and Secretion
Authors: JENSEN TONE SKEIE , BJØRGE LINE , WOLLEN ANNE‐LONE , ULSTEIN MAGNAR
Published: May 2013

PROBLEM: Complement lytic activity has been demonstrated, and a potential for its activation is present in human cervical and tubal secretions and in the endometrium. This necessitates the presence of regulatory mechanisms for protection of the sperm and the implanting allogeneic conceptus in the female genital tract. Complement regulatory proteins demonstrated on sperm and in seminal fluid have been attributed such a role. It is however likely that additional protection is required for a successful conception and implantation to take place. This lead us to investigate the distribution of the complement regulatory factors in cervical mucus and mucosa, uterine endometrium, and fallopian tube.

METHOD: Endometrium and cervical mucosa were obtained from patients undergoing hysterectomy for benign conditions, and specimens were selected from different stages of the menstrual cycle. Fallopian tubes were obtained from patients submitted for sterilization, while cervical mucus was aspirated from volunteers undergoing gynecological examination. Immunohistochemistry was performed on all tissue samples, using monoclonal antibodies to membrane cofactor protein (MCP), decay accelerating factor (DAF), CD59 and complement receptor 1 (CR1). Western blot analysis was performed on cervical mucus under nonreducing conditions.

RESULTS: MCP, DAF, and CD59 were found to be expressed in human endometrium and fallopian tube. No variation in expression was detected throughout the menstrual cycle. CR1 was not expressed. Soluble forms of DAF and CD59 were found to be present in cervical mucus.

CONCLUSION: The complement regulatory proteins MCP, DAF, and CD59 are expressed throughout the female genital tract, and may thus play an important role in protecting the traversing sperm and implanting blastocyst from complement mediated damage.