bluuebunny/crossref_metadata_2025 huggingface dataset:
Rat Pregnancy Immune‐regulatory Circuits Are Progestation Hormonal Status, Decidual Tissue, Embryo‐Trophoblast and Late Pregnancy Changes Dependent
Authors: Mohammad Mohammad , Saadi May , Balhaa Ghassan Rai El , Chateaureynaud Pierrette , Mayer Guy
Published: May 2013

ABSTRACT: We have tested the spleen cells of rats for their reactivity to mating and unrelated (third party) strain alloantigens during pregnancy, pseudopreg‐nancy, and traumatic deciduoma installation. By cytotoxic T‐lymphocyte assays, after 7‐day one‐way mixed lymphocyte cultures (MLC), we detected the presence of many immunoregulatory circuits among rat pregnancy and postpartum. They are progestation hormonal status, decidual tissue, embryo‐trophoblast, late‐pregnancy‐parturition‐associated changes, and lactation‐hormonal‐status dependent.
Monoclonal Antibodies Against Human Chorionic Gonadotropin (hCG): II. Affinity and Ability to Neutralize the Biological Activity of hCG
Authors: BERGER PETER , KOFLER REINHARD , WICK GEORG
Published: May 2013

ABSTRACT: Monoclonal antibodies (MCA) against hCG have been characterized with regard to their affinity and their ability to neutralize the biological activity of hCG in vivo. The production and specificities of these reagents were described in the preceeding paper of this series. Equilibrium association constants (Ka) of the MCA, determined by radioimmunological saturation assays, ranged from less than 1 × 108M−1 up to 3.7 × 109M−1 whereas values for conventional polyclonal antisera against hCG ranged from 8.9 × 109M to 1.8 × 1010M−1. The ability of MCA to neutralize the biological activity of hCG was tested in a rat bioassay in vivo; 9 of 13 different MCA preparations tested could neutralize hCG. Surprisingly, this property did not correlate with affinity or specificity, and was not restricted to those MCA recognizing the hormone specific β‐subunit. It could be demonstrated that determinants on each individual subunit as well as epitopes formed by both subunits are involved in the expression of the biological activity of hCG.
Immunoregulatory Function of Human Cord Blood Lymphocytes on Immunoglobulin Production
Authors: CHENG HENRY , SEHON ALEC H. , DELESPESSE GUY
Published: May 2013

ABSTRACT: The functional maturity of human umbilical cord blood B lymphocytes and the immunoregulatory activity of cord T lymphocytes were assessed by measuring the in vitro immunoglobulin production by B cells from either cord or adult blood. Supernatants from 48‐hr pokeweed‐mitogen (PWM) stimulated cord or adult lymphocyte cultures were added to cord or adult B cell cultures in the presence of PWM; a significant amount of immunoglobulin was produced in adult B cell cultures only. Adult B or T cells were then cocultured with cord T or B cells; a significant amount of immunoglobulin was again found only in adult B cell cultures. These results indicated that cord B cells were functionally immature and that cord helper T cell function was adequate but masked by excessive suppressor activity. Indeed, addition of cord T cells but not of allogeneic adult T cells to PWM stimulated adult lymphocyte cultures inhibited their immunoglobulin production; this confirmed cord T cells' increased suppressor activity. Cord T cells were not intrinsically suppressive since they failed to suppress immunoglobulin production by Epstein‐Barr Virus (EBV) transformed B cells. They could be activated, however, by PWM or allogeneic cells (in mixed lymphocyte cultures) and their effect was mediated via soluble factor(s) as demonstrated by the suppressor effect of these culture supernatants on immunoglobulin production by unfractionated adult lymphocytes. In contrast, when these supernatants were added to T cell‐depleted adult lymphyocyte cultures, enhancement rather than suppression was observed. These results indicated that the soluble factor(s) released by Cord T lymphocytes was not suppressing per se but induced suppression through activation of suppressor cells.
A Simplified Method Using Enzyme‐Linked Immunosorbent Assay for Titration of Antisperm Antibodies
Authors: ZANCHETTA R. , BUSOLO F.
Published: May 2013

ABSTRACT: We selected sera from 44 patients (36 males and 8 females) that were positive for antisperm antibodies using ELISA (titer ranged from 1:32 to 1:512) for the evaluation of a simplified method for determination of antisperm antibodies. This method uses a correlation between a single absorbant value and the endpoint titer of the same serum. This simplified procedure increases the number of the sera that can be tested on each plate, resulting in considerable saving of time, reagent costs, and materials. A standard curve allows the direct determination of endpoint titer using the absorbance value found at a single dilution. This modification improves the utility of the assay for the epidemiological screening of antisperm antibodies in patients who may have an immunological cause of infertility.
Role of Histoincompatibility on the Blastogenic Response of Lymphocytes From Guinea Pigs Alloimmunized With Sperm Cells
Authors: NEMIROVSKY MARIO S.
Published: May 2013

ABSTRACT: The involvement of antigens coded by the major histocompatibility complex in alloimmunization of guinea pigs with sperm has not been fully explored. Some of the “in vitro” correlates of cell‐mediated immunity, such as the lymphocyte transformation test in presence of sperm cells, could be influenced, at least partially, by histoincompatibility. Outbred Hartley and inbred strain 13 guinea pigs were alloimmunized with sperm cells emulsified in Freund's complete adjuvant. The degree of histocompatibility between recipients and sperm cell donors was determined by using mixed lymphocyte culture assays. Stimulatory indices higher than four, indicating histoincompatibility, were obtained frequently between members of Hartley strain or between Hartley and strain 13 guinea pigs, but never between strain 13 animals. Sperm cells from histoincompatible guinea pigs failed to stimulate blastogenesis in lymphoid cells from nonimmunized animals. The lymphoblastogenic potential of allo‐ and isospermatozoa were compared. Lymphoid cells from Hartley and strain 13 guinea pigs alloimmunized with sperm cells were incubated in vitro with histocompatible and histoincompatible sperm cells. Both strain 13 and Hartley sperm cells gave a similar stimulatory index with lymphoid cells belonging to outbred or inbred alloimmunized guinea pigs. The use of fresh spermatozoa was of paramount importance. Frozen specimens lost their stimulatory capacity almost completely. Within the limits of the technique utilized here, we have failed to detect any influence of the antigens coded by the major histocompatibility complex of the guinea pig upon the sperm‐induced lymphocyte transformation. These observations point to the sperm‐specific autoantigens as being responsible for the lymphocyte proliferation.
Experimental Autoimmune Damage to Rat Male Accessory Glands. I. Transfer of Autoimmune Response by Spleen Cells
Authors: DEPIANTE‐DEPAOLI M. , PACHECO‐RUPIL B. , BRITOS S. , CASAS A.
Published: May 2013

ABSTRACT: Spleen cells obtained 30 days after the first immunization of rats isoimmunized with an extract of male accessory glands (MAG) were capable of adoptively transferring specific cell‐mediated immunity to normal syngenic recipients. Humoral and cell‐mediated immune responses were investigated in donor and in the recipient rats that were killed 7 days after intravenous (IV) injection of the cells. In recipient rats the cell‐mediated immunity showed multiple ways of expression and in some cases this was exemplified by a sharp increase in regard to the donor's response. Furthermore, a widespread reactivity in the spleen, lymph nodes, and thymus cells was detected. On the contrary, no circulating antibodies to MAG antigens were demonstrated after the spleen cell transfer. Cell separation studies showed that a nylon woolnonadherent cell was responsible for the transfer of the cell‐mediated immune response. This was abrogated by depletion of T lymphocytes and treatment with antirat thymocytes serum and complement. The mechanism of transfer and development of the cell‐mediated immunity in recipient rats is discussed.
Lymphocyte Subpopulations in Normal and Preeclampsia Pregnancies
Authors: GUSDON JOHN P. , HEISE EUGENE R. , QUINN KATHY J. , MATTHEWS LINDA C.
Published: May 2013

ABSTRACT: We have made an effort to determine whether or not there is any change in subpopulations of lymphocytes in normally pregnant and preeclamptic pregnancies using monoclonal antibody markers. Eleven normally pregnant and ten women with preeclampsia were studied, both during the third trimester and again two months postpartum, and compared to eleven age‐matched nonpregnant women. Mononuclear cells were isolated from heparinized venous blood. One million cells were treated with each appropriate antibody (Ortho‐mune OKM1, OKT3, OKT4, OKT8, OKT11, OKIa), and then reacted with FITC‐antimouse IgG and examined by flow cytometry and/or fluorescence microscopy. No significant differences between these three groups were noted in the OKT3, OKT4, OKT8, OKT11, or OKIa cellular populations. The OKM1 population was significantly decreased in the third trimester of normal pregnancies but not in the preeclamptic pregnancies. No significant differences were found 2 months postpartum.
Effects of Immunization of Mothers on the Immune Reaction of Their Offspring: Inhibition of Immune Responses of Offspring Caused by Antibody Imported Through the Milk
Authors: IZUCHI KUNIO , TANIGUCHI KAZUTO , GONDO HISASHI , KAWAUCHI HIDEYUKI , KUBO CHIHARU , SHIRAKAWA KOUICHI , NOMOTO KIKUO
Published: May 2013

ABSTRACT: Effects of immunization of pregnant AKR mice with nucleated chicken erythrocytes (CRBC) on immune responses of their offspring were examined. Antigen‐specific reduction of generation of cytotoxicity and plaque forming cells (PFC) was demonstrated in the offspring at 8 weeks after birth, and lasted for 15 weeks. Cross‐fostering experiments and cell transfer experiments showed that such suppression would be induced by antibody contained in the milk of immunized mothers rather than suppressor cells. Activities to enhance opsonization and to mediate antibody‐dependent cell‐mediated cytotoxicity (AI)CC) were demonstrated in the serum of such offspring before challenge with CRBC. Delayed footpad reaction (DFR) was maintained at the normal level in such offspring of immunized mice.
Clinical Experience With the Prevention of Rh‐lsoimmunization: A Historical Comparative Analysis
Authors: KEITH LOUIS , DANIS RONALD P. , BERGER GARY S.
Published: May 2013

ABSTRACT: Clinical experience with prevention of Rh‐immunization is reviewed. The pathogenesis of hemolytic disease and the chemistry of prophylaxis is explained. The factors which effect antigenic expression are delineated. The clinical indications for prevention of AMIS are reviewed. International data pertaining to Rhesonativ are presented from postpartum trials and antepartum trials. A protocol for antepartum administration of anti‐Rh immunoglobulin is given.
Alpha‐Fetoprotein Subtractions in Germ Cell Tumors Identified by Con A or LCH Crossed‐Line Affinity Immunoelectrophoresis
Authors: ISHIGURO TATSUYA , SAWADA MASUMI , SAKAGUCHI HIDEO , SUGITACHI ICHIBEI
Published: May 2013

ABSTRACT: Using concanavalin A (Con A) crossed‐line affinity immunoelectrophoresis and lentil lectin (LCH) crossed‐line affinity immunoelectrophoresis, alpha‐fetoprotein (AFP) subfractions were studied in sera including three sera from nude mice heterotran‐splanted with human yolk sac tumor of the ovary and three sera from patients with yolk sac tumor, mature solid teratoma, or immature solid teratoma of the ovary. In sera of nude mice bearing yolk sac tumor or from a patient with yolk sac tumor, subfractions from yolk sac and those from fetal liver were identified. Since AFP subfractions from yolk sac and fetal liver can be differentiated according to the carbohydrate moieties, our findings indicate that AFP produced by yolk sac tumor and fetal yolk sac are to some extent differently glycosylated. We also found that AFP in both mature and immature solid teratoma was composed of two subfractions ontogenetically originating from yolk sac or fetal liver. All these findings indicate that more than two different factors are responsible for the AFP synthesis in germ cell tumor of the ovary.
Vasectomy‐lnduced Autoimmunity: Antisperm and Antinuclear Autoimmune Monoclonal Antibodies
Authors: ISAHAKIA MOHAMED , ALEXANDER NANCY J.
Published: May 2013

ABSTRACT: Antibodies to sperm and testicular autoantigens are a hallmark of vasectomy‐induced murine autoimmunity. We generated five autoimmune monoclonal antibodies with spleen cells from nonimmunized but vasectomized BDF1 male mice as fusion partners in two experiments. Four of the antibodies, designated Vx 4, Vx 8, Vx 10, and Vx 23, recognize, as determined by fluorescence microscopy, topographically restricted sperm‐specific antigenic determinants that appear in the germinal epithelium as spermatogenesis progresses. Antibodies Vx 8 and Vx 10 identified 60,000‐ and 97,000‐dalton, mouse‐specific sperm antigens restricted to the anterior acrosome and posterior acrosome, respectively. Antibody Vx 23 identified a 35,000‐ to 40,000‐dalton antigen on the midpiece and postacrosomal regions of epididymal mouse sperm. The antigenic target of antibody Vx 4 on the midpiece region could not be identified with the Western blot technique. Both Vx 4 and Vx 23 cross‐reacted with human, rabbit, and monkey sperm; immunofluorescence revealed that they were bound onto the midpiece region. By immunofluorescence, a single clone (Vx 24) distinctly reacted with the nuclei of somatic and testicular germ cells but not the nuclei of sperm. The isolation of this antinuclear autoantibody raises questions about the development of nonsperm‐specific autoantibodies after vasectomy in the mouse.
Monoclonal Sperm Antibodies: Their Potential for Investigation of Sperms as Target of Immunological Contraception
Authors: METTLER LISELOTTE , PAUL SUDHIR , BAUKLOH VERA , FELLER ALFRED C.
Published: May 2013

ABSTRACT: We generated 149 hybridoma cell lines secreting antibodies against human spermatozoal antigens of which antibodies from 136 hybridoma lines also reacted with seminal plasma constituents. The occurrence of common antigeneic determinants on spermatozoa and seminal plasma was further demonstrated by competitive inhibition ELISA tests. We found that seven hybridoma clones secreted antibodies reactive with spermatozoa but nonreactive with seminal plasma. Antibodies from 5 clones were sperm‐agglutinating and from 15 clones sperm‐immobilizing. Localization of sperm antigens reactive with the monoclonal antibodies was demonstrated by indirect immunoperoxidase staining. A synthetic decapeptide, earlier shown to be reactive with naturally occurring human iso‐ and autosperm antibodies, was shown to be reactive with the monoclonal antibody VII‐5 in ELISA tests.
Relationship of Antibodies to Sperm Head to Etiology of Infertility in Patients Undergoing In Vitro Fertilization/Embryo Transfer
Authors: MANDELBAUM STACEY LEE , DIAMOND MICHAEL P. , DECHERNEY ALAN H.
Published: May 2013

ABSTRACT: Antibodies to sperm head (ASA‐H) are believed to impair reproduction, probably because of a reduction in fertilization of human oocytes. However, the incidence of ASA‐H in couples with different etiologies of infertility undergoing in vitro fertilization/embryo transfer (IVF/ET) is unestablished. To examine this question, the semen, serum, and follicular fluid of 11 couples with unexplained infertility and 25 couples with tubal infertility undergoing IVF/ET were tested with the immunobead binding assay to identify ASA‐H of IgA, IgG, and IgM isotypes. Comparing couples with unexplained vs. tubal infertility, 46% vs. 4% had ASA‐H of at least one isotype in female serum (P = .006), 36% vs. 4% had ASA‐H in follicular fluid (P = .023), 27% vs. 0% had ASA‐H in semen (P = .023), and 18% vs. 4% had ASA‐H in male serum (P = .022), respectively. ASA‐H were present in one or more fluids tested in 55% of patients with unexplained infertility, compared to 8% of patients with tubal infertility (P = .005). Of the six women with ASA‐H in their serum, 83% (5/6) were undergoing IVF/ET for unexplained infertility compared to 17% (1/6: P = .08) undergoing IVF/ET for tubal infertility. In summary, clinically significant ASA‐H are present in a substantial number of infertile women undergoing IVF/ET, particularly those whose infertility is unexplained. Based on these findings, we conclude that it is efficacious to screen all women with unexplained infertility undergoing IVF/ET for ASA‐H.
Treatment of Recurrent Spontaneous Abortion by Immunization With Paternal Lymphocytes: Correlates With Outcome
Authors: GATENBY PAUL A. , MOORE HELEN , CAMERON KAYE , DORAN TREVOR J. , ADELSTEIN STEPHEN
Published: May 2013

ABSTRACT: Previous observations have suggested that defective recognition of fetal alloantigens by the maternal immune system is associated with recurrent pregnancy failure and that this may be prevented by boosting the maternal immune system with paternal or pooled third‐party leukocytes. The mechanism whereby this process achieves success is not clear, and accordingly to explore this we immunized 28 couples with recurrent fetal loss with 80 × 106 paternal peripheral blood mononuclear leukocytes (PBML) and followed various immunological parameters. The couples studied, in whom 55% achieved a successful pregnancy, showed no increase in sharing of human lymphocyte antigen (HLA)‐A, ‐B, or ‐DR antigens and no consistent evidence of a decreased mixed leukocyte reaction (MLR) or MLR plasma‐blocking factors compared with control couples. Immunization did not alter these parameters but did induce antipaternal lymphocytotoxins, although the presence of the latter did not correlate with pregnancy outcome. There was a correlation between rapid conception after immunization and a subsequent successful pregnancy. A successful pregnancy also correlated with sustained postimmunization, postconception maternal antipaternal allospecific CD‐8+ suppressor T cells. Although these findings provide overall evidence that immunization produces changes in the way in which the maternal immune system interacts with the fetus, larger numbers of couples and a higher dose of paternal lymphocytes will be needed to establish clearly whether this therapy works and its mechanism of action.
Experimental Autoimmune Oophoritis. I. Inhibition of Fertility in Rats Isoimmunized With Homogenates of Ovary
Authors: DAMJANOVIĆ A , JANKOVIĆ BRANISLAV D.
Published: May 2013

ABSTRACT: Experimental autoimmune oophoritis (EAOO) induced in female Lewis rats by a single injection of isologous ovary in complete Freund's adjuvant (CFA) and concentrated pertussis vaccine resulted in a multifocal accumulation of mononuclear cells in ovaries and complete loss of fertility after 17–28 days. The time course of appearance of anti‐ovary antibodies and their serum titers showed a good correlation with the reduction of litter size. Furthermore, the infertility also could be induced by passive immunization of rats with adsorbed anti‐ovary serum. The contraceptive effect of the antibody does not result in permanent sterility. Besides lymphoid cell infiltrations, the most prominent histological alterations in ovaries of experimental rats were characterized by a significant increase in the number of atretic preantral and antral follicles and a decrease in the number of growing follicles of all sizes and corpora lutea. It is presumed that both the direct effect of anti‐ovary antibodies and disturbance in ovarian morphology can lead to a reduction of litter size and appearance of infertile cycles in rats with oophoritis.
Suppression of T‐Lymphocyte Blastogenesis by Ovine Trophoblast Protein‐1 and Human Interferon‐α May Be Independent of Interleukin‐2 Production
Authors: NIWANO YOSHIMI , HANSEN THOMAS R. , KAZEMI MOHAMMAD , MALATHY PUTHAN‐VEEDU , JOHNSON HAROLD D. , ROBERTS R. MICHAEL , IMAKAWA KAZUHIKO
Published: May 2013

ABSTRACT: Cells derived from the trophoblast tissue of a day 15 sheep conceptus released substances that inhibit incorporation of [3H]thymidine into phytohemagglutinin (PHA)‐stimulated ovine lymphocytes. This effect was partially reversed by addition of antiserum to ovine trophoblast protein‐1 (oTP‐1), a major secretory product of day 13–21 sheep conceptuses and a protein structurally and functionally related to α‐interferons (IFN‐α). Human IFN‐a, unlike dexamethasone, inhibits phytohemagglutinin‐induced lymphocyte blastogenesis without reducing interleukin‐2 (IL‐2) production by the cultures, and conditioned medium containing IL‐2 does not promote [3H]thymidine incorporation into ovine lymphocytes when oTP‐1 is present. Thus, oTP‐1, by virtue of being an IFN, may have a local immunomodulatory role by selectively inhibiting the proliferative responses of certain maternal immune cells to IL‐2.
Antiphospholipid Antibodies and Recurrent Pregnancy Loss
Authors: TRIPLETT DOUGLAS A.
Published: May 2013

ABSTRACT: Antiphospholipid antibodies (APA) have recently been linked to a variety of clinical findings including arterial and venous thrombosis, recurrent spontaneous abortions and fetal loss, as well as thrombocytopenia. These observations have stimulated multidisciplinary interest in the laboratory identification of APA and the clinical management of patients with positive tests for APA.
Functional Heterogeneity of Sialic Acid Binding Agglutinins of Rat Uteri Towards In Vitro Lymphocyte Transformation
Authors: BHATTACHARYYA INDRANI , MANDAL CHITRA , CHOWDHURY IDULA
Published: May 2013

ABSTRACT: The sialic acid binding agglutinins (SAS) are purified from three different stages (proestrus [P], estrus [E], diestrus [D]) of rat uterine homogenate. All three proteins are capable of inhibiting the in vitro phytohemagglutinin (PHA)‐induced blastogenic response of lymphocytes and thymocytes as manifested by inhibition of 3H‐thymidine incorporation into cells. The lymphocytes isolated from pregnant female rats were found to be more susceptible to inhibition. In resting lymphocyte cultures (RLC), however, the P‐and E‐SAS proteins acted differently from the D‐SAS by stimulating lymphocyte proliferation in the cultures initiated from normal rat lymphocytes. At the same dose these proteins were inhibitory to the RLC, where cultured lymphocytes were isolated from the pregnant female. Reverse hemolytic plaque formation assay results also indicated functional heterogeneity between these proteins. P‐SAS induced a potent polyclonal activation of the B cell, whereas E‐ and D‐SAS did not show much activation. SAS proteins do not have any complement‐like activity.

These results, corroborated by cell viability count and 51Cr uptake tests, suggest that D‐SAS has genuine lymphoblastogenesis‐suppressive properties whereas P‐ and E‐SAS‐induced suppression in the presence of PHA is probably due to an increased cell‐density‐dependent cytotoxicity.
Monoclonal Antibodies to Human Secretory “Pregnancy‐Associated Endometrial α1‐Globulin,” an Insulin‐Like Growth Factor Binding Protein: Characterization and Use in Radioimmunoassay, Western Blots, and Immunohistochemistry
Authors: BELL STEPHEN C. , JAMES ROGER F.L. , JACKSON JULIET A. , PATEL SHILPA R. , WAITES GILLIAN T. , WALCZAK KAREN
Published: May 2013

ABSTRACT: Monoclonal antibodies were raised against pregnancy‐associated endometrial ax‐globulin (α1‐PEG), a 32 KD insulin‐like growth factor binding protein (IGF‐BP), which represents a major secretory product of the human decidualized endometrium during pregnancy. This class of IGF‐BP has been implicated in the modulation of action, inhibitory and stimulatory, of insulin‐like growth factors. Immunization with the protein purified from pregnancy endometrium resulted after myeloma fusion in the isolation of six hybridoma clones and the antibodies produced were characterized. The Ka of the antibodies ranged between 4.75 × 109 M1 and 0.7 × 108 M−1. In Western blots all monoclonal antibodies reacted with purified protein of molecular weight 32 KD and specifically detected this IGF‐BP species in culture medium and cytosolic extracts of pregnancy endometrium and amniotic fluid. The monoclonal antibodies appear to define three epitope‐bearing regions as evidenced by their reactivity to polypeptide fragments of the protein. After synthesis and secretion by tissue explants in vitro the protein is susceptible to cleavage into fragments possessing different monoclonal antibody‐defined reactivity. Employing immunohistochemical techniques the protein was principally localized to decidual cells in tissue sections of pregnancy endometrium and solely to these cells after enzymic digestion of the tissue. The implications of these results are discussed with respect to potential role of IGF‐BP in the action of IGF upon the IGF‐1 receptor‐bearing populations, including lymphocytes and trophoblast cells, D in the decidua.
Factors of the Extrinsic Pathway of Blood Coagulation in Fetal Placental Macrophages
Authors: KAPPELMAYER JÁNOS , ÁDÁNY RÓZA
Published: May 2013

ABSTRACT: Term human placentae were examined to identify and localize the factors contributing to extravascular fibrin formation. In addition to blood vessels, components of the extrinsic coagulation cascade were also demonstrated in intracellular localization. These cells in double and treble labeling systems expressed macrophage marker antigens, recognized by DAKO antimacrophage, RFD7, Amersham antimacrophage, and KiM7 monoclonal antibodies and showed positivity for alpha‐naphthyl‐acetate‐esterase (ANAE). The fact that factors of the extrinsic coagulation system can be demonstrated in fetal macrophages of the chorionic stroma suggests that their role is not restricted to cellular defense and phagocytosis, but they may be involved in extravascular intraplacental fibrin formation as well.
MLR‐Blocking Antibodies Are Directed Against Alloantigens Expressed on Syncytiotrophoblasts
Authors: GOTO SHIGENORI , TAKAKUWA KOICHI , KANAZAWA KOJI , TAKEUCHI SHOSHICHI
Published: May 2013

ABSTRACT: In this study, the reactivity of mixed lymphocyte reaction‐blocking antibodies toward trophoblasts was investigated. Nineteen sera that had a significant level of MLR‐blocking effect were obtained from normal pregnant women and from primary habitual aborters after immunotherapy. The blocking effects of these sera were studied before and after absorption with syncytiotrophoblast plasma membrane (STPM) from the respective mothers' placenta (autologous) as an experimental group and from unrelated mothers' placenta (allogeneic) as a control. The HLA types of cord blood lymphocytes corresponding to ten of these STPM were then determined.

In the study with the 11 normal pregnancy sera, the blocking effect of every serum was partially removed by autologous STPM absorption. This revealed a significant difference between the preabsorption and postabsorption MLR‐blocking effect. Nevertheless, absorption by the allogeneic STPM either partially removed or did not remove the blocking effect. Furthermore, there was a statistically significant difference in the elimination of the MLR‐blocking effect between the autologous STPM and the allogeneic STPM. Studies with the sera from the immunized aborters revealed similar results. There was no statistically significant difference in the elimination of the antibodies between absorption with autologous and with allogeneic STPM. In these habitual aborters' sera, comparison of paternal stimulator and cord blood lymphocytes HLA types showed that the positive allogeneic STPM absorptions could not be explained sufficiently by coincidence of HLA types. Further studies are needed to determine whether the alloantigens are HLA‐linked or not.
Alteration in Platelet Count During Early Pregnancy in the Mouse
Authors: ELIAS K.A. , DAS A.K. , PARDUE D. , COULAM C.B. , CRITSER J.K. , CRITSER E.S.
Published: May 2013

ABSTRACT: Published reports of pregnancy associated thrombocytopenia in mice have utilized the Quackenbush strain. The inability of some laboratories to verify this observation in other mouse strains prompted us to report our findings by using Swiss Albino ICR mice. In Exp. 1, pregnant and pseudopregnant mice were bled prior to mating (time 0) and daily on day 1 (vaginal plug) through day 7. In Exp. 2, media from 24 hr cultures of 2‐cell mouse embryos or media from unfertilized oocytes were injected into splenectomized mice. Animals were bled at time 0 (before injection) and at 30,60, and 120 min after injection. In Exp. 3, splenectomized mice were treated with either media from 2‐cell stage embryos or with media supplemented with synthetic platelet‐activating factor (PAF: 0.05, 0.1 or 0.2 μg). Animals were bled as in Exp. 2. Platelet numbers were determined in duplicate from each blood sample by using a hemacytometer. In Exp. 4, antagonist (SRI 63–441) or vehicle was administered to mated mice on days 1 through 4 of pregnancy. Animals were examined on day 8 to determine number of developing conceptuses. In Exp. 1–3, data were analyzed by using ANOVA for repeated measures, and in Exp. 4 data were analyzed by chi‐square analysis. In Exp. 1, there was a treatment × time interaction (P<.06) due to transient thrombocytopenia in pregnant but not pseudopregnant mice. In Exp. 2, there was a significant treatment × time interaction (P<.02) with platelet numbers being markedly reduced at 30 min in animals treated with media from 2‐cell embryos but not with media from unfertilized oocytes. In Exp. 3, the response to PAF was not different (P>.10) than the response to 2‐cell embryo culture media. In Exp. 4, a total dose of 20 μg of antagonist did not alter (P>.1) the frequency of animals pregnant on day 8 while 20 μg/day of antagonist significantly (P<.05) reduced the frequency of animals that were pregnant. These data agree with reports using Quackenbush mice and support the hypothesis that reduction in platelet numbers is associated with early pregnancy in mice.
Pathology of Intrauterine Growth Retardation
Authors: YEE KHONG T.
Published: May 2013

ABSTRACT: Uteroplacental vasculature is described and resultant placental pathology in intrauterine growth retardation (IUGR) discussed. The historical basis for suggesting an immunological role in IUGR is reviewed. Recent developments in cytogenetic studies that may have relevance in the IUGR are presented.
HLA Expression on Human Ejaculated Sperm
Authors: OHASHI KAZUTOMO , SAJI FUMITAKA , KATO MUNEHIRO , WAKIMOTO AKINORI , TANIZAWA OSAMU
Published: May 2013

ABSTRACT: The expression of human leukocyte antigen (HLA) on human ejaculated sperm has been investigated by using cellular binding radioimmunoassay (CB‐RIA) with sperm samples purified by Percoll discontinuous gradient and monoclonal antibodies specific to the frame work of HLA molecules. The results in packed sperm obtained from ten healthy males indicated that HLA class II and its subclasses, DP, DQ, and DR, were expressed at a low level on human ejaculated sperm, while HLA class I was not detected. Furthermore, we could not detect the individual variation of HLA expression on ejaculated sperm in the cases of twenty healthy males.

K562, which is B lymphoid cell line and lacks HLA class I molecules, showed detectable amounts of HLA class I molecules after exposure to interferon gamma (IFN‐γ) and/or tumor necrosis factor alpha (TNF‐α). However, the treatment of sperm with IFN‐γ and/or TNF‐α had no effect on the expression of HLA class I and class II molecules.
Progesterone Suppression of Pregnancy Lymphocytes Is not Mediated by Glucocorticoid Effect
Authors: SZEKERES‐BARTHO JULIA , PHILIBERT D. , CHAOUAT G.
Published: May 2013

ABSTRACT: This study investigated whether the suppressive effect of progesterone on pregnancy lymphocytes is mediated by specific progesterone receptors. The effects of a competitive progesterone antagonist (RU486) and a specific glucocorticoid receptor blocker (RU43044) were tested on the release of a blocking factor by progesterone‐treated pregnancy lymphocytes. RU 486 tested at an equal concentration as progesterone significantly inhibited the production of the blocking factor, while RU 43044 was without effect. These data suggest that in pregnancy, lymphocyte progesterone acts on specific progesterone receptors and glucocorticoid binding sites are not involved.
A Murine Model of NK Cell Mediated Resorption
Authors: KINSKY RADSLAV , DELAGE GENEVIÉVE , ROSIN NATHALIE , THANG MING NGUY , HOFFMANN MIREILLE , CHAOUAT GÉRARD
Published: May 2013

ABSTRACT: There is increasing evidence that some models of immunologically mediated murine embryo demise involve nonspecific lytic effector cells. In this paper, we use two double stranded synthetic RNAs, known as potent interferon inducers and NK cell activators, the Poly (I).Poly (C) and the less toxic Poly (I).Poly (C12U). These polynucleotides enhance fetal resorption rates in both resorption prone and none‐resorption prone strains of mice. We have studied the kinetics of the phenomenon, and observed an anti‐implantation‐like effect of early injection during early pregnacy. The abortifacient effects can be adoptively transferred to naive recipients by spleen cells from Ds RNA injected donors. Such effects are abrogated if the cells are pretreated with anti‐NK cell antiserum. The relevance of these findings to the survival of the conceptus is suggested.
Identification of an Oolemmal IgG Fc Receptor: Its Role in Promoting Binding of Antibody‐Labelled Human Sperm to Zona‐Free Hamster Eggs
Authors: BRONSON R.A. , FLEIT H.B. , FUSI F.
Published: May 2013

ABSTRACT: Antisperm antibodies (ASAs) present in sera of infertile men and women have been shown either to promote or inhibit penetration of zona‐free hamster eggs by antibody‐labelled human spermatozoa. Increased numbers of oolemmal‐bound sperm have been noted in association with increased sperm penetration frequencies, following antibody labelling, when compared with antibody‐free sperm. The promotion of adherence of ASA‐labelled sperm to the oolemma could be mediated through the binding of antibodies to common epitopes present on the sperm and egg surfaces or through Fc‐mediated binding to an oolemmal Fc receptor. In support of the latter hypothesis, we report that zona‐free hamster eggs bind aggregated human IgG and IgG Fc fragments. The presence of an oolemmal IgG Fc receptor has been confirmed using a rat monoclonal antibody (2.4G2) directed against a murine IgG Fc receptor (Fc γ RII) as judged both by indirect immunofluorescence and by immunobead binding. In addition, the pre‐incubation of zona‐free hamster eggs with IgG Fc diminished both adhesion to and penetration of the oolemma by human spermatozoa.
Expression of Class II Major Histocompatibility Complex Antigen on Mouse Sperm and Its Roles in Fertilization
Authors: MORI TSUNEATSU , WU GUO MAO , MORI ETSUKO , SHINDO YUTAKA , MORI NORIKO , FUKUDA AISAKU , MORI TAKAHIDE
Published: May 2013

ABSTRACT: The expression of class II major histocompatibility complex (MHC) antigen on the membrane of mouse sperm head was clearly demonstrated by indirect immunofluorescence (IIF) test, enzyme immunoassay (EIA), and mixed lymphocyte sperm reaction (MLSR) as well as the examination of its expression at transcriptional levels by nothern dot blotting. Furthermore, the roles of class II MHC antigen of sperm in fertilization were investigated with an in vitro fertilization (IVF) system. The successful ratio of IVF was significantly decreased by treatment of sperm with anticross‐reactive region of class II MHC antigen monoclonal antibody (MAb) but not with antiprivate region MAb. These results were not due to disturbances of sperm mobility by these MAbs. It was strongly suggested that the monomorphic or its related region of class II MHC antigen on sperm played an important role in the recognition between sperm and egg in fertilization.
Cell Surface Antigen Expression of First Trimester Chorionic Villus Samples
Authors: LATA JENNIFER A. , COWCHOCK F. SUSAN , JACKSON LAIRD G. , SMITH J. BRUCE
Published: May 2013

ABSTRACT: First trimester chorionic villi obtained by chorionic villus sampling at approximately 9 weeks of gestation were investigated by indirect immunofluorescence to demonstrate trophoblast cell surface antigen expression. Villous trophoblast expressing the trophoblast specific markers transferrin receptor, human placental lactogen, and cytokeratin was also found to express a monomorphic major histocompatibility complex class I determinant recognized by the monoclonal antibody W6/32. W6/32 positive regions included sparsely scattered regions of villous trophoblast and fanning outgrowths of trophoblast. The class I antigenic determinant expressed by first trimester trophoblast was found to be recognized exclusively by W6/32 when assayed with a panel of anti‐class I determinant monoclonal antibodies. Trophoblast W6/32 determinant expression was not increased after 24 hour organ culture in the presence of 200 U of interferon gamma. Exposure to interferon gamma resulted in increased class I antigen expression by mesenchyme and low level de novo mesenchyme class II antigen expression. These data suggest that early gestational stage villous trophoblast express non‐classical class I antigens which do not seem to be subject to the regulatory effects of interferon gamma.
Major Histocompatibility Complex Class I Antigens Expressed on Rat Trophoblast Cells
Authors: SAITO MITSURU , MISRA DHIRENDRA N. , KUNZ HEINZ W. , GILL THOMAS J.
Published: May 2013

ABSTRACT: There is controversy about the size of the major histocompatibility complex antigens of trophoblast cells from placenta. There are some reports that the heavy chains of these molecules are smaller (39–43 kd) than those of the classical class I antigens (45–46 kd), while there are others which show that both the light and the heavy forms of class I antigens occur in the trophoblast cells. In order to investigate this problem, we studied the classical class I antigen (RT1.Aa) and the pregnancy‐associated class I antigen (Pa) of the rat from 125I‐labeled basal trophoblast cells, isolated from the placenta of WF females pregnant by DA males, using very mild conditions. These antigens were compared with those of the syngeneic (DA × DA) trophoblast cells or paternal (DA) lymphocytes by sodiun dodecyl sulfate‐polyacry‐lamide gel electrophoresis. Both the Aa and Pa antigens, precipitated from the two trophoblast preparations, showed a heavy chain of 46 kd associated with a 12 kd (β2‐microglobulin component, as did the same molecules precipitated from the lymphocytes. Heavy chains in the range of 39–43 kd could not be detected in any of the samples. The results suggest that the smaller molecular weight heavy chains are methodological artifacts and could arise from loss of a glycan(s) during isolation.
Characterization of Autoantigens Relevant to Experimental Autoimmune Orchitis (EAO) in Mice Immunized With a Mixture of Syngeneic Testis Homogenate and Klebsiella O3 Lipopolysaccharide
Authors: YOKOCHI TAKASHI , IKEDA HIDEO , INOUE YOSHIKO , KIMURA YOSHINOBU , ITO HIDENOBU , FUJII YASUAKI , KATO NOBUO
Published: May 2013

ABSTRACT: Experimental autoimmune orchitis (EAO) in mice has been induced by repeated injection of a mixture of syngeneic testis homogenate and Klebsiella O3 lipopolysaccharide (KO3 LPS) as a potent adjuvant. The antisera obtained from mice with EAO lesions defined several antigens with apparent molecular weights (MW) of 38,000 (38 kd), 86 kd, 100 kd, and > 200 kd by the immunoblotting method. These antigens were organ‐specific and exclusively present on the acrosome of spermatozoa, suggesting that these acrosomal antigens were highly relevant to EAO. It was found that the antigen with a fairly high MW (> 200 kd) was expressed on spermatozoa from the epididymis. Furthermore, the acrosomal 86 kd antigen was predominantly expressed in the testis, while the 100 kd antigen was dominant in the spermatozoa from the epididymis. It was therefore suggested that the 86 kd and 100 kd antigens in the acrosome were differentially expressed on the process of maturation of spermatozoa.
Influence of Blood Monocytes and Lymphocytes on Progesterone Production by Granulosa Cells From Small and Large Follicles in the Pig
Authors: STANDAERT FRANK E. , CHEW BOON P. , WONG TERI S.
Published: May 2013

ABSTRACT: The objectives of this study were 1) to determine the influence of lymphocytes and monocytes on progesterone secretion by porcine granulosa cells in culture, and 2) to compare the responses of granulosa cells obtained from small versus large follicles. Granulosa cells were cocultured with lymphocytes, monocytes, and unfractionated mononuclear cells in serum‐free culture medium. Progesterone content of media and cellular DNA were determined after 6, 12, and 24 hrs of culture. Progesterone production by granulosa cells alone or in coculture with erythrocytes increased (P < .001) up to 12 hrs and plateaued thereafter. Cells from large follicles produced approximately two‐fold (P < .01) more progesterone as compared to those isolated from small follicles. Cell numbers, as indicated by DNA, did not change with time in any of the cultures. Lymphocytes stimulated (P < .05) progesterone production by granulosa cells from large follicles but not from small follicles. Similar results were seen when unfractionated mononuclear cells were co‐cultured with granulosa cells. Monocytes stimulated progesterone production by granulosa cells from both large and small follicles. Lymphocytes and monocytes influence steroid secretion by granulosa cells in the pig in vitro. These effects are not mediated through cell proliferation and are influenced by follicle size.
Detection of Immunoregulatory Lipid‐Like Factors in Human Amniotic Fluid
Authors: RUEDA RICARDO , VARGAS MA LUISA , GARCÍA‐PACHECO MARCOS , GARCÍA‐OLIVARES ENRIQUE
Published: May 2013

ABSTRACT: The immunosuppressive activity of amniotic fluid (AF) is extensively documented in the mouse. Although this property is due in part to the presence of alpha‐fetoprotein (α‐FP), other immunosuppressive factors are suspected. In this article, we demonstrate that human amniotic fluid lipid extract (AFLE) is inhibitory of, although not cytotoxic to, PHA‐activated human lymphocytes, of mouse bone marrow cells, and of different established cell lines of human and mouse source. This effect is shown to be reversible. Under preparative thin layer chromatography (TLC) using chloroform:methanol:water (60:38:8) as solvents, the activity of AFLE migrates to two peaks of inhibition with Rf values of 0.46‐0.62 and 0.84‐1, respectively. These lipid‐like factors may play a role as a nonspecific immunoregulatory mechanism which prevents maternally mediated immune rejection of the conceptus.
Danazol Suppresses the Production of Interleukin‐1β and Tumor Necrosis Factor by Human Monocytes
Authors: MORI HIDEHIRO , NAKAGAWA MIKI , ITOH NAOKI , WADA KEISUKE , TAMAYA TERUHIKO
Published: May 2013

ABSTRACT: The effects of estradiol (E2), progesterone (P), and danazol on the production of interleukin‐1β (IL‐1β) and tumor necrosis factor (TNF) by OK‐432 (a streptococcal preparation)‐stimulated monocytes were examined. E2 and P at physiologic concentrations enhanced IL‐1β and TNF production by monocytes from donors with lower control levels (without steroids added) of IL‐1β and TNF. However, E2 and P at physiologic concentrations did not affect IL‐1β and TNF production by monocytes from donors with higher control levels of IL‐1β and TNF. Danazol inhibited IL‐1β and TNF production by monocytes in a dose‐dependent manner from not only donors with lower control levels of IL‐1β and TNF but also donors with higher control levels of IL‐1β and TNF. Danazol at a concentration of 10−6 M significantly suppressed IL‐1β and TNF production in the presence of E2 and/or P at concentrations giving peak responses of IL‐1β production. These findings suggest possible new mechanisms of action for danazol in the treatment of endometriosis and infertility associated with immune abnormalities.
Studies on T‐Lineage Cells in Human Decidua of First Trimester Pregnancies
Authors: SATO SHIGEYASU , KANZAKI HIDEHARU , YOSHIDA MASUMI , TOKUSHIGE MAKOTO , WANG HUEI‐SHENG , KARIYA MASATOSHI , OKAMOTO NORIHIKO , KARIYA YOSHITAKA , UCHIDA ATSUSHI , KASAKURA SHINPEI , MORI TAKAHIDE
Published: May 2013

ABSTRACT: T‐lineage cells in human decidua of early pregnancies were tested for surface markers, proliferative response, interleukin‐2 (IL‐2) production, and natural killer (NK) activity. T‐lineage (CD2+) cells that were obtained from decidua by the use of E‐rosette formation contained fewer CD3+ mature T cells and CD4+ cells than those from the peripheral blood of the same donors, while no differences were seen in the frequencies of CD8+ cells. P55 molecules of IL‐2 receptor (IL‐2R/p55, Tac antigen) were hardly detected on fresh decidual T‐lineage cells, though approximately 20% were positive for HLA‐DR. More than a half of decidual T‐lineage cells expressed CD56 molecules on their surface and killed K562 cells, the prototype target of NK cells, while most of them were negative for CD16 and CD57. Upon stimulation with IL‐2, decidual T‐lineage cells demonstrated dose‐dependent proliferative response. In addition, they were induced to produce high amounts of IL‐2 by stimulation with mitogens but not with alloantigens. These results suggest that human decidua contains high numbers of CD2+3‐CD16+56+ lymphocytes and that this population responds to IL‐2, produces IL‐2 and mediates NK activity.
Ubiquitous Expression of TNF‐α/Cachectin Immunoreactivity in Human Endometrium
Authors: TABIBZADEH S.
Published: May 2013

ABSTRACT: Emerging evidence suggests that cytokines play significant roles as intercellular communication signals in human endometrium. In the present report, an immunohistochemical staining method and tumor necrosis factor‐α (TNF‐α) specific polyclonal antibody was used to identify potential in vivo sources of this cytokine in human endometrium. During the proliferative and early secretory phases of the menstrual cycle, glandular epithelium was distinctly free of any immunoreactive product. With the emergence of mid‐ and particularly late‐secretory phases, however, some glandular epithelial cells started to express TNF‐α. In the late secretory phase, glands exhibiting strong staining were adjacent to those that were negative for TNF‐α or only sparingly contained positively immunostained cells. Remarkably, the glandular epithelium constituted a major source of enhanced expression of TNF‐α during gestation. In the proliferative phase, virtually all stromal cells strongly expressed TNF‐α, and in the secretory phase, some variability of staining could be observed among various stromal cells. In gestational endometria, decidual cells and in all endometria, endothelial cells strongly expressed immunoreactivity for TNF‐α. These data demonstrate that various constituents of endometria are constitutively primed to express TNF‐α and that this expression is distinctly associated with the changes in endometrium that are compelled by hormonal stimuli.