bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunohistochemical Localization of Interferons in Human Placental Tissues in Normal, Ectopic, and Molar Pregnancy
Authors: BULMER JUDITH N. , MORRISON LYNN , JOHNSON P.M. , MEAGER A.
Published: May 2013

ABSTRACT: Interferon (IFN)α, β, and γ have been localized in normal and pathological human pregnancy using both polyclonal and monoclonal antibodies in immunohistochemical techniques. IFNα was localized to fetal chorionic villous syncytiotrophoblast throughout normal pregnancy, as well as to extravillous trophoblast in the placental bed and chorion lave. Maternal decidual leukocytes, as well as fetal Hofbauer cells in the villous mesenchyme, also contained IFNα. IFNγ was detected in villous syncytiotrophoblast, while anti‐IFNβ showed only patchy weak reactivity with syncytiotrophoblast. Reaction patterns on ectopic pregnancy tissues were similar to those in early intrauterine pregnancy. In molar pregnancy, reactivity for IFNα, β, and γ was observed in syncytiotrophoblast. Along with their potential anti‐viral effects, placental interferons could play a role in local immunomodulation or in regulation of embryonic cell proliferation and differentiation.
Mononuclear Cell Subpopulations in Human Follicular Fluid From Stimulated Cycles
Authors: CASTILLA JOSE A. , SAMPALO ALMUDENA , MOLINA RAMON , SAMANIEGO FRANCISCO , MOZAS JUAN , VERGARA FRANCISCO , GARRIDO FEDERICO , HERRUZO ALFONSO J.
Published: May 2013

ABSTRACT: The study of lymphocyte subsets from human follicular fluid (FF) provides an opportunity to evaluate immunological features of the ovary. We investigated the mononuclear cell subsets in FF and peripheral blood obtained at the time of laparoscopy from ten in vitro fertilization (IVF) patients. Midcycle nonpregnant peripheral blood was used as the control. A marked increase in the proportion of monocytes (CD14+) was observed in FF. Although FF was enriched with CD8+ lymphocytes, a decrease in the proportion of CD4+ lymphocytes was observed. “Memory” T cells in FF, identified by the CD4+ CD45R− phenotype, predominated over “naive” T cells (CD4+ CD45R+) at a ratio of 2:1, which differs from the ratio yielded by control blood samples (1:1). The percentage of activated T cells (CD3+ HLA‐DR+ cell) increased significantly in FF. When lymphocyte subsets were studied in the peripheral blood of IVF patients, changes similar to but less significant than those in FF were found. These data support the concept that lymphocytes play an important role in ovarian physiology.
Humoral Immunity to Candida albicans (Anti‐Candida Antibody Titers) in Premature Infants
Authors: KATIKANENI LAKSHMI D. PAPPU , MATHUR SUBBI
Published: May 2013

ABSTRACT: Although the role of humoral and cell mediated immunity in neonatal defense against Candida infections is not precisely defined, one of the contributing immunologic factors may be a lack of decreased specific passive humoral immunity. Thus, serum samples from the umbilical veins of 98 term gestation and 105 premature neonates (majority <33 wk gestation) and their mothers (n = 100) were tested for the presence of hemagglutinating antibodies to commercially available Candida antigen. The titers of Candida antibodies (mean log2 ± SEM) were significantly higher in 11 term neonates (4.73 ± 0.69) of mothers with high antibody titers (5.18 ± 0.40, <0.001) as contrasted with 87 normal term (2.38 ± 0.15) and 105 premature (2.87 ± 0.15) infants with normal mothers (1.96 ± 0.13 and 3.31 ± 0.26, respectively). Contrary to our belief 81% of term infants and all of the preterm infants (majority <33 wk gestation) had passive specific anti‐candida antibody titers <1:16.
Human Trophoblast Cells in Culture Express an Unusual Major Histocompatibility Complex Class I‐Like Antigen
Authors: GRABOWSKA ANNA , CARTER NIGEL , LOKE Y.W.
Published: May 2013

ABSTRACT: We present evidence that the trophoblast cells obtained from first trimester placentae by a culture method developed in our laboratory express a class I‐like HLA antigen that is only sparsely distributed on the cell surface, lacks classical polymorphic determinants, and consists of heavy chains of lower molecular weight than those of classical HLA in association with β2‐microglobulin (β2M). The latter two features are in general agreement with a previous report describing the nature of the HLA molecule on trophoblast disaggregated from term amniochorion. These cultured trophoblast cells could be useful for molecular studies of the unusual class I antigen, as well as for other in vitro experiments designed to mimic the in vivo local trophoblastuterine interactions.
Lymphocyte‐Derived Progesterone‐Induced Blocking Factor Corrects Resorption in a Murine Abortion System
Authors: SZEKERES‐BARTHO JULIA , CHAOUAT G.
Published: May 2013

ABSTRACT: An anti‐abortive effect of a progesterone‐induced blocking factor has been shown in the CBA/J × DBA/2 abortion system. I.p. injections of pregnant mice on days 7.5, 9.5, and 11.5 of gestation with dialyzed supernatants from progesterone‐treated murine pregnancy spleen cells (but not from control ones) significantly (P< 0.01) reduced the resorption rate from 47.7% in the untreated animals to 20.7%.
Hydatidiform Mole Pregnancy Trophoblast Extracts Differentially Suppress lnterleukin‐2‐lnduced Proliferation of Human T‐Lymphocytes and PHA‐Blasts
Authors: BENNETT WILLIAM A. , BRACKIN MARTHA N. , MCGEHEE RAMON P. , COWAN BRYAN D.
Published: May 2013

ABSTRACT: Immunoregulatory factors of trophoblast origin may partially abrogate maternal immune responses to the fetus during pregnancy. We have previously shown that soluble factors extracted from hydatidiform mole trophoblast suppress interleukin‐2 (IL‐2)‐dependent proliferation of a cloned murine cytotoxic T cell line (CTLL‐2). To characterize human T cell responses to this trophoblast extract, we measured the effects of molar tissue extracts (HME) on IL‐2‐stimulated proliferation of human T‐lymphocytes and mitogen (PHA) transformed T‐cell blasts (PHA‐blasts). HME significantly (P<0.05) suppressed T‐lymphocyte proliferation in response to 5 and 10 units/ml of IL‐2 at 500 and 250 μg/ml, while no effect was observed at the 100 μg/ml concentration. Suppression by HME of IL‐2‐stimulated T‐cell proliferation was partially overcome by the addition of excess IL‐2. HME also suppressed (P<0.05) IL‐2‐stimulated proliferation of PHA‐blasts at 500 and 250 μg/well at both 5 and 10 units/ml of IL‐2. As observed with resting T‐cell responses, no suppression of PHA‐blast proliferation was observed using 100 μg/ml of HME. In contrast to the response of the resting T‐cells to excess IL‐2, HME suppression of IL‐2‐stimulated blast proliferation was not affected by increasing the concentration of IL‐2. These results indicate that extracts from hydatidiform mole trophoblast contain immunosuppressive factors that block human T‐cell clonal expansion by inhibiting the utilization and/or production of IL‐2. Furthermore, the effects of HME are not reversed by excess IL‐2 when PHA‐blasts are reacted compared to resting T‐cell responses, which are partially reversed in the presence of excess IL‐2. This suggests that human trophoblast‐derived immunoregulatory factors that are present in hydatidiform mole inhibit human lymphocyte utilization of IL‐2 in a fashion similar to the murine (CTLL) model, and exhibit irreversible effects on lymphocytes containing high affinity IL‐2 receptors (PHA‐blasts) compared to lymphocytes with only low affinity IL‐2 receptors (resting T‐cell). This property of the trophoblast may partially explain how hydatidiform mole, and possibly pregnancy trophoblast in general, escapes lethal immunological challenge by the maternal immune system.
Morphologic Changes in the Rat Uterus Following Natural Mating and Embryo Transfer
Authors: KANBOUR‐SHAKIR AMAL , KUNZ HEINZ W. , GILL THOMAS J. , ARMSTRONG DAVID T. , MACPHERSON TREVOR A.
Published: May 2013

ABSTRACT: In order to gain some insight into the putative immune suppression that may be induced at the placental implantation sites, the morphological changes at these sites following natural matings and following the transfer of embryos fertilized in vivo were studied. The only histologic parameter that showed a significant difference was the number of granulated metrial gland (GMG) cells. More GMG cells were present in allogeneic than in syngeneic pregnancies, and more GMG cells were present following embryo transfer into an allogeneic female than following the comparable natural mating. The role of the GMG cells in pregnancy is, however, still unresolved.
Lack of Complement Activation in the Seminal Plasma of Men With Antisperm Antibodies Associated In Vivo on Their Sperm
Authors: D'CRUZ OSMOND J. , HAAS GILBERT G.
Published: May 2013

ABSTRACT: A specific and sensitive “sandwich”‐type radiolabeled antiglobulin assay (RAA) using monoclonal anti‐human C5b‐9 neoantigen and polyclonal anti‐human C5b‐9 was used to evaluate the presence of the in vivo product of human complement (C) activation (SC5b‐9) in the seminal plasma (SP) of 19 fertile and 61 infertile men. SP SC5b‐9 was detectable in 7 (8.7%; 1 fertile and 6 infertile men) of the 80 men with a range of 10 to 175 ng/ml. Levels of SP SC5b‐9 in other men were below the limit of detection (less than 10 ng/ml). Of the 33 infertile men with sperm‐associated immunoglobulin (Ig) G and/or IgA, 27 (82%) had undetectable levels of SP SC5b‐9 immunoreactivity. There was no correlation between the SP SC5b‐9 levels and the degree of sperm‐associated IgG (r = 0.086) or IgA (r = 0.23) activity. However, significant deposition of sperm‐bound C5b‐9 due to autologous C activation was demonstrated by flow cytometry of donor sperm treated with sera from autoimmune men with ASA in their sera and on their sperm. These findings suggest that sperm‐bound Ig cannot activate C in SP.
Differential Modulation of the In Vitro Lymphocyte Activation Pathways by Soluble and Solubilized Placental Substances
Authors: DUC HUYNH THIEN , NAKAGAWA SHOKO , RUCAY PIERRE , RIGHENZI SIMONE , VOISIN GUY‐ANDRÉ
Published: May 2013

ABSTRACT: Soluble and detergent‐solubilized placental extracts were studied for their modulatory effects upon the proliferation of lymphocytes stimulated by various activating agents. It was shown that soluble placental extract (SPE) exerted an inhibitory effect on the lymphoproliferation triggered by alloantigen or LPS but not by Con A or the combined action of PMA + calcium ionophore A 23187. This effect was also observed with SPE precipitated by 30% of ammonium sulfate (SPE30). On the other hand, a solubilized placental extract (SzPE) that was obtained by using octyl‐β‐D‐glucopyranoside inhibited the stimulation triggered by alloantigen, LPS, and Con A but did not affect the protein kinase C pathway. The modulatory effects were observed not only when SPE (or SPE30) and SzPE were added at the time of culture initiation but also at 24 h before or after the activating agents. Preincubation with SPE30 or SzPE immobilized on plastic surface, however, transduced an enhanced lymphoproliferative response to alloantigen and mitogen Con A but not to LPS. The above results suggest that placental substances exerted their modulatory effects by interfering mainly with the antigen or mitogen lymphoproliferation pathways.
Immune Modulation of Natural Killer Cell Cytotoxicity Against Herpes Infected Target Cells in Pregnancy
Authors: GONIK BERNARD , LOFTIN KARIN C. , TAN NENITA S. , CRUMP JERRY
Published: May 2013

ABSTRACT: Natural killer cell cytotoxicity (NKC) is a nonspecific, primary immunodefense system active against a variety of pathogens, including herpes simplex virus (HSV). Evidence suggests that during pregnancy, NKC is attenuated. The regulatory mechanisms for this immune attenuation have yet to be defined. We examined two cytokines (interleukin‐2 [IL‐2] and alpha interferon [IFN]) for their ability to alter NKC responsiveness during pregnancy, utilizing an HSV‐infected target cell model. Peripheral mononuclear effector cells were isolated from 19 pregnant and 19 nonpregnant subjects by Ficoll‐Paque separation. These cells were incubated with IFN, IL‐2, or media alone, and analyzed for %NKC by an 18 h chromium release assay. The percentage of NKC was lower using the effector cells from the pregnant subjects as compared to nonpregnant controls. Incubation with either IFN or IL‐2 resulted in a significant augmentation of NKC in both the pregnant and nonpregnant derived cells. There were no differences in IL‐2 dose requirements or levels of cytotoxicity achieved (43.1 ± 6.8% vs. 44.4 ± 6.8%, respectively) between pregnant and nonpregnant derived cells. The IFN‐mediated augmentation of NKC was somewhat blunted in pregnancy both in terms of absolute levels of cytotoxicity achieved (26.1 ± 3.9% vs. 37.2 ± 4.9%, respectively) and dose response curves generated. These results demonstrate that NKC against HSV infected cells is attenuated during pregnancy and can be immunoregulated with the use of either IFN and IL‐2. The restoration of NKC responsiveness with IFN, however, remains incomplete during pregnancy, suggesting that this cytokine's mechanism of action differs from that of IL‐2.
Familial Recurrent Spontaneous Abortions
Authors: MOWBRAY JAMES F. , UNDERWOOD JENNIFER , GILL THOMAS J.
Published: May 2013

ABSTRACT: Recurrent spontaneous abortions can occur as a familial trait—for at least three generations—in some families.
Antinuclear Autoantibodies in Healthy Nonpregnant and Pregnant Women and Their Offspring
Authors: PIURA BENJAMIN , TAUBER ERNEST , DROR YAARA , SAROV BATIA , BUSKILA DAN , SLOR HANOCH , SHOENFELD YEHUDA
Published: May 2013

ABSTRACT: Antinuclear autoantibodies have previously been detected in sera of healthy women although less frequently than in sera of women with autoimmune disorders. The effect of pregnancy on antinuclear autoantibody production in healthy women is as yet debatable. We present four studies in which, by employing the ELISA method, we evaluated the presence of six antinuclear autoantibodies (anti‐ds DNA, anti‐ss DNA, anti‐poly(I), anti‐cardio‐lipin, anti‐Sm, and anti‐RNP) in the sera of more than 1,000 healthy pregnant and nonpregnant women, including 196 pairs of matched maternal and cord blood sera. In all four studies healthy pregnant women did not demonstrate significantly higher prevalence rates of various serum antinuclear autoantibodies as compared to healthy nonpregnant women. All detected autoantibodies were of the IgM isotype. In only one infant (born to a healthy seronegative mother) was an autoantibody (IgM anti‐ss DNA) detected. This may indicate that in certain circumstances the fetus is capable of self‐production of autoantibodies.
Antisperm Antibodies in Split Ejaculates
Authors: MEINERTZ HELLE
Published: May 2013

ABSTRACT: The split ejaculation technique concentrates the most motile and viable spermatozoa in the first part of the ejaculate. Several clinicians use this fraction for insemination and in vitro fertilization. Eleven vasovasostomized men with antisperm antibodies participated in this study. The mixed antiglobulin reaction (MAR) for IgG, IgA, and secretory component was carried out on the whole ejaculate and the fractions of the split ejaculate. The isotype and concentration of free antisperm antibodies were determined in serum, the whole ejaculate and the fractions of the split ejaculate by the indirect MAR and the tray agglutination test. The detection of bound antisperm antibodies revealed almost identical reactions in fractions one and two and in the whole ejaculate. The results suggested that the majority of antisperm antibodies in the ejaculate from vasovasostomized men are transuded from serum not only at the epididymal and/or the prostatic level but also in the seminal vesicles.
Suppression by Human Placental Protein 14 of Natural Killer Cell Activity
Authors: OKAMOTO NORIHIKO , UCHIDA ATSUSHI , TAKAKURA KENJI , KARIYA YOSHITAKA , KANZAKI HIDEHARU , RIITTINEN LEENA , KOISTINEN RIITTA , SEPPÄLÄ MARKKU , MORI TAKAHIDE
Published: May 2013

ABSTRACT: Human decidua of early pregnancy contains considerable numbers of CD3− CD56+ natural killer (NK) cells. In this study, two major protein products of the decidua, placental protein 14 (PP14) and placental protein 12 (PP12), were tested for the ability to regulate human NK cell activity. In vitro overnight exposure to PP14 of blood lymphocytes or purified large granular lymphocytes (LGL) resulted in suppression of cytotoxicity against K562 target cells in a 4‐h 51Cr release assay. The NK inhibition was dependent on concentrations of PP14, being detectable at 5 μg/ml and reaching maximum at 50 μg/ml. Manifestation of PP14‐induced NK suppression required 18‐h contact with NK cells. The suppression of NK activity by PP14 was not abolished by indomethacin. In a target binding assay the number of PP14‐treated LGL binding to K562 was comparable to that of untreated ones. By contrast with PP14, PP12 produced no effects on NK cells. These results indicate that PP14 suppresses the function of NK cells, which might be involved in prevention of maternal immune rejection of fetus at the fetomaternal interface.
Positive Immunoselection—A Method of Isolating Leukocytes From Leukocytic Reacted Human Cervical Mucus Samples
Authors: THOMPSON L.A. , TOMLINSON M.J. , BARRATT C.L.R. , BOLTON A.E. , COOKE I.D.
Published: May 2013

ABSTRACT: In this brief communication we report a simple and accurate method of isolating and quantifying specific leukocytes from midcycle human cervical mucus, using monoclonal antibody‐coated magnetic beads. Cervical mucus samples (pre‐ and postinsemination) were broken down enzymatically and incubated with a series of these beads. This method of positive immunoselection consistently retrieved representative levels of leukocytes (x̄ = 73.8% ± 1.59%; mean leukocyte retrieval rate ± S.E.) from the cervical mucus samples. Significantly more leukocytes (P< 0.0001) were isolated from the postinsemination samples, the predominant leukocyte of which was the neutrophil, which comprised 83% of the leukocyte population. These results reaffirm that a leukocytic influx is initiated across the human uterine cervix following the introduction of semen samples, the function of which is possibly phagocytic clearance of the nonfertilizing population of sperm.
Monitoring of Smooth Muscle Antibodies in Hypertensive Complications During Pregnancy
Authors: PIETARINEN ILMO , HARTIKAINEN‐SORRI ANNA‐LIISA , KIVINEN SEPPO , MÄKITALO REINO
Published: May 2013

ABSTRACT: Smooth muscle antibodies (SMA) were monitored in terms of titer, immunoglobulin class and staining pattern by indirect immunofluorescence assay during the pregnancies of 32 women with pregnancy induced hypertension (PIH), 21 with essential hypertension (EH), and 43 with a normal pregnancy. The mean SMA titer was higher both in PIH and EH than in normal pregnancy. SMA were mainly of the IgG class and gave a “vessel wall” staining pattern in immunofluorescence. The tendency for the mean SMA titer to increase was observed in PIH, whereas a falling tendency was found in EH and normal pregnancy. These tendencies suggest that hypertension, the duration of which has been shorter in PIH, induces SMA production. The observed staining pattern suggests the same kind of non‐actin specificity for SMA. as in our previous study of complications in early pregnancy.
The Lack of Direct Effects of a Monoclonal Antibody Against Murine T‐Cell Suppressor Factor on Murine Embryo Development In Vitro
Authors: HOVERSLAND R.C. , BEAMAN K.D.
Published: May 2013

ABSTRACT: In previous studies, we reported that the injection of monoclonal antibody 14–30, specific for a T‐cell suppressor factor (TSF), into mice during early stages of pregnancy could decrease the percentage of females that maintained pregnancy. In addition, further work has demonstrated the presence of an immunoreactive protein in fetal and maternal tissues with physiochemical properties similar to TSF. However, one alternate explanation for the antipregnancy effects of the injections of monoclonal antibody, not related to a specific role for TSF in early pregnancy, is the possibility of direct effects upon the embryo or embryonic antigens that prevent continued embryonic development. In the present studies, early preimplantation embryos were incubated with the monoclonal antibody 14–30 specific for TSF and the subsequent development of the embryos examined. The results of these studies demonstrate that monoclonal antibody, which has been shown to bind T‐cell suppressor factor and has antipregnancy effects when injected in vivo, does not interfere with development of preimplantation or implantation stage mouse embryos in vitro.
Binding of Sperm‐Reactive Antibodies in Human Sera to Surface‐Associated Antigens on Human Sperm Compared by Indirect Immunobead, Immunofluorescence, and Immunogold Assays
Authors: YOUNG L. G. , SMITHWICK E. B.
Published: May 2013

ABSTRACT: Human sera were identified as positive or negative for sperm‐reactive antibodies in a solid‐phase enzyme‐linked immunosorbent assay (ELI‐SA). Of these, 28 positive and 22 negative sera were blind‐coded and used as first antibody to compare three immunoassays, a modified liquid‐phase indirect immunobead assay (IRA); a liquid‐phase indirect immunofluorescence assay (IFA); and a solid‐phase indirect immunogold assay (IGA). These three immunoassays perform both as sperm‐reactive antibody detection assays and as sperm‐associated antigen localization assays. As antibody detection assays, the IBA, IFA, and IGA gave 37, 27, and 28 positives and 13, 23, and 22 negatives, respectively. The usefulness of the IBA as an antigen localization assay was limited by the size of the marker, while the smaller IFA and IGA markers enabled increased resolution of binding patterns of sperm‐reactive antibodies to surface‐associated sperm antigens. Although the antigen‐antibody binding patterns were almost identical for IFA and IGA, suggesting the same sperm‐associated antigens were detected by both assays, the IGA reaction product was stable, higher in resolution, and visible by light microscopy.
Effect of Sperm‐Antibodies on Acrosome Reaction of Human Sperm Used for the Hamster Egg Penetration Assay
Authors: FRANCVILLA FELICE , ROMANO ROSSELLA , SANTUCCI RICCARDO
Published: May 2013

ABSTRACT: The effect of anti‐sperm antibodies (ASA) on the rate of acrosome reactions (AR) during “in vitro” capacitation of human sperm used for the hamster egg penetration assay (HEPA) was assessed. Motile sperm suspensions from donors were exposed to several sera and seminal plasma with sperm head‐directed ASA, then they were washed and capacitated “in vitro.” After capacitation, the proportion of acrosome‐reacted viable sperm was assessed by staining with Fluoresceinated Pisum Sativum Agglutinin and supravital stain Hoechst 33258. ASA of any immunoglobulin class did not significantly affect either the AR rate, or the hamster egg penetration rate. In conclusion, interference of ASA on spontaneous AR rate during “in vitro” capacitation can not be advocated as an explanation of the impairment of the interaction of human sperm with egg or its vestments, which have been reported in several studies.
Investigations Concerning the Relationship of Ovarian Eosinophilia to Ovulation and Luteal Function in the Sheep
Authors: MURDOCH W. J. , STEADMAN L. E.
Published: May 2013

ABSTRACT: Eosinophilic leukocytes are attracted into the preovulatory follicle and regressing corpus luteum of sheep. Eosinophils produce a wide array of potent bioactive substances that could play a role in ovarian function. Circulatory eosinophils were depleted by treatment of ewes with prednisolone, thereby producing ovarian eosinopenia. Ovulation occurred in these animals. However, circulatory concentrations of progesterone during the ensuing luteal phase were less than normal. This insufficiency was attributed in part to a suppressed angiogenic response in the formative corpus luteum. Regression of the corpus luteum induced by prostaglandin F2α was not affected by administration of prednisolone. Thus, an obligatory role of eosinophils in the mechanics of ovulation and luteal regression in the sheep is unlikely.
Evidence in Support of the Plasma Membrane as the Target for Transferrin‐Adriamycin Conjugates in K562 Cells
Authors: BARABAS KLARA , SIZENSKY JOSEPH A. , FAULK W. PAGE
Published: May 2013

ABSTRACT: We have used transferrin‐adriamycin conjugates to deliver drug to transferrin receptors and have shown that the conjugates bind to and kill tumor cells in vivo and in vitro. This has been studied with the use of qualitative and quantitative assays. In this report, we present evidence indicating that the primary target of transferrin‐adriamycin cytotoxicity is the plasma membrane. This was done using a spectrofluorometric method that takes advantage of the fluorescence properties of adriamycin. It was shown that, while DNA intercalation may be the primary mechanism of cytotoxicity for free adriamycin, transferrin‐adriamycin conjugates were not observed to interact with nuclear DNA. This may be a useful consideration in the design of future chemotherapeutic studies with transferrin conjugates of anticancer drugs.
Trophoblast Immunity in Human Pregnancy Defined by Antiidiotype
Authors: TORRY DONALD S. , FAULK W. PAGE , MCINTYRE JOHN A.
Published: May 2013

ABSTRACT: Successful reproduction in mammals requires the mother to immunologically accept genetically disparate tissues. Allotypic trophoblast antigens (TLX) are thought to be responsible for influencing maternal acceptance of the feto‐placental graft, and faulty regulation of immunity to TLX antigens has been associated with recurrent pregnancy losses. In this report, rabbit antiidiotype (RAb2) was produced to a human TLX antibody (Ab1). This RAb2 detected TLX cross‐reactive idio‐types (CRI) on antitrophoblast IgG from women with normal and abnormal pregnancies. These findings support an hypothesis that women respond immunologically to allotypic trophoblast antigens, and that idiotype‐antiidiotype regulation of this response is characteristic of normal pregnancy.
The Leukocytic Reaction of the Human Uterine Cervix
Authors: THOMPSON L.A. , BARRATT C.L.R. , BOLTON A.E. , COOKE I.D.
Published: May 2013

PROBLEM: Sperm interaction with the immune system of the human cervix is poorly understood.

METHOD: The leukocytic response of the human cervix to sperm was examined in a closely monitored patient population (N = 10), using monoclonal antibody cell identification techniques. Baseline data were collected from both cervical mucus and smears sampled before treatment by donor insemination. Donor insemination was timed to coincide with ovulation by monitoring plasma LH concentrations twice daily. Following insemination the numbers of leukocytes were recorded in cervical mucus and smear samples taken over a 24‐h period relative to the time of treatment. Controls treated with “pure sperm,” seminal plasma, cryopreservative, and cervical smearing alone were also included in the study.

RESULTS: Only those women treated with sperm cells exhibited substantial elevations in leukocyte numbers following inseminations. Additionally, serial cervical smearing induced an inflammatory response of the cervix. In all the women, the neutrophil was the predominant leukocyte of the cervix both during the baseline and treatment periods (median values ranged from 77 to 86%). Macrophages, T‐helper lymphocytes, and T‐suppressor lymphocytes were also detected, but only in low numbers (2–10.6%). Two patients and one control (“pure sperm”) became pregnant during their study cycle.

CONCLUSIONS: We conclude that the leukocytic reaction is a physiological response of the cervix to sperm, the function of which remains to be established.
Immunotherapy of Habitual Abortion
Authors: CARP H.J.A. , TODER V. , MASHIACH S.
Published: May 2013

ABSTRACT: Many cases of habitual abortion have been assumed to be due to hyporesponsiveness to the spouse's antigens encountered in pregnancy. Immunization by paternal leukocytes has been used to potentiate the immune response and prevent further miscarriages. This treatment has been highly controversial in terms of efficacy, mode of action, and side effects. More recently immunoglobulin has been used as passive immunization for similar indications.

In our experience immunotherapy is effective; 80% of patients have subsequent live births. The most significant results are seen in patients with five or more abortions, in whom 66% of subsequent pregnancies develop normally compared to 20% in a control group. We have used antipaternal complement‐dependent antibody (APCA) production after immunization as a marker of immune response. APCA correlates with beneficial outcome in the next pregnancy. APCA may also be associated with cytokines, which may enhance embryonic and trophoblast development. Immunoglobulin may similarly provide the relevant antibodies or cytokines. At present a large scale meta‐analysis is being performed to confirm or refute the efficacy of this treatment This meta‐analysis may resolve the controversy.
Immunological Factors in Infertility
Authors: SHUSHAN ASHER , SCHENKER JOSEPH G.
Published: May 2013

ABSTRACT: Immune infertility can result from destruction of gametes by antisperm antibodies (ASA) or anti‐ovary antibodies, by inhibition of sperm‐zona pellucida binding by ASA, or by prevention of embryo cleavage and early development. Condoms, immunosuppressive therapy, sperm processing, and intrauterine insemination have been widely applied, but with controversial results.
Stable Production of Recombinant Human Sperm Immobilizing Antibody Using cDNA Expression Vectors
Authors: SAWAI HIDEAKI , YAMASAKI NORIYUKI , KOMORI SHINJI , KARASUYAMA HAJIME , ISOJIMA SHINZO
Published: May 2013

PROBLEM: Sperm immobilizing antibodies present in sterile women may be one of the principal causes of immunological infertility. We already established cell lines that secrete recombinant human IgG sperm immobilizing antibody using class‐switched (from IgM to IgG) genomic immunoglobulin genes. However, these transfectants produced a small quantity of antibody and required continuous use of a medium with selective reagents. We have now constructed cell lines that stably secrete the antibody in large quantities using immunoglobulin cDNAs and cDNA expression vectors.

METHOD: The immunoglobulin heavy chain cDNA was cloned from transfectants that secrete the class‐switched human IgG sperm immobilizing antibody. The light chain cDNA, which had already been cloned, and the heavy chain cDNA were inserted into the modified bovine papilloma virus‐based cDNA expression vectors BCMGSNeo and BCMGSHyg, respectively. These constructs were sequentially transfected into a mouse myeloma cell line by electroporation.

RESULTS: The established transfectants produced recombinant antibody that retained human sperm immobilizing activity in nonselective medium for at least 30 days. Moreover, the production of the antibody was increased three times over that of the previous cell lines.

CONCLUSION: We have established an unique method that improves the production of sperm immobilizing antibody stably and in large quantities.
Antiendometrial Autoantibodies Are Generated in Patients With Endometriosis
Authors: GORAI ITSUO , ISHIKAWA MASAHIKO , ONOSE RYO , HIRAHARA FUMIKI , MINAGUCHI HIROSHI
Published: May 2013

PROBLEM: Our aim was to investigate endometrial antigens involved in the autoimmunity of endometriosis.

METHOD: We detected endometrial antigens against which autoantibodies directed with Western blotting.

RESULTS: Thirteen (72.2%), 14 (77.8%), and 15 (83.3%) of 18 serum samples from endometriosis patients had antibodies reactive against endometrial antigen wtih MW of 26 kd, 34 kd, and 42 kd, respectively, while 6 (33.3%), 8 (44.4%), and 8 (44.4%) of 18 samples from normal control women reacted against these antigens, respectively. The frequencies of antibodies to the endometrial antigens were significantly (P < 0.05) higher in the endometriosis patients than in the normal control women. Antibodies in peritoneal fluid (PF) reacted against antigens with MW of 26, 34, 38, 42, and 64 kd, while those from the normal control reacted against antigens wtih MW of 38, 42, and 64 kd. Serum samples from normal fertile males did not show any reactivity against these endometrial antigens.

CONCLUSIONS: Our results show that autoantibodies reactive against endometrial antigens are present in patients with endometriosis.
Mitogen Induced Production of Polyclonal IgG is Decreased in Women With Severe Endometriosis
Authors: GEBEL HOWARD M. , BRAUN DONALD P. , ROTMAN CARLOS , RANA NASIR , DMOWSKI W. PAUL
Published: May 2013

PROBLEM: The etiology and/or pathogenesis of endometriosis may involve aspects of both humoral and cellular immunity.

METHOD: In this investigation, we analyzed the ability of B lymphocytes from distinct patient groups for production of IgGl, IgG2, and IgG3 following in vitro stimulation with polyclonal B‐cell mitogens (pokeweed mitogen and Staphylococcus aureus Cowan strain I) after in vitro stimulation with polyclonal B‐cell activators.

RESULTS: We observed that the in vitro production of IgGl, IgG2, and IgG3 was identical among fertile controls (no endometriosis; N = 22), infertile women without endometriosis (N = 22), infertile women without endometriosis (N = 20) and patietns with stage 1 or 2 endometriosis (N = 31). In contrast, in vitro IgG2 production was significantly reduced among women with stage 3 or 4 endometriosis (N = 11) compared to controls (P < 0.001).

CONCLUSION: Since the number of circulating B cells was similar in each patient group studied, the reduced production of IgG2 in patients with stage 3 or 4 disease was not merely due to fewer antibody producing cells in those subjects, and we speculate that the observed decrease in polyclonal IgG2 production among these patients is due to a primary defect. In additional studies, we observed that polyclonal IgG2 production was normal among stage 3 or 4 patients treated with danazol (N=ll), but significantly reduced in patients treated with gonadotropin releasing hormone agonists (N = 8). Although not conclusive, these data suggest that danazol may have the capacity to correct the defective production of polyclonal IgG2 in patients with severe endometriosis.
A Solid‐Phase Assay for the Detection of Anti‐Sperm Antibodies
Authors: OKADA HIROSHI , KAMIDONO SADAO , OWENS GEORGE R. , NAGAMATSU R. GEORGE , ADDONIZIO JOSEPH C.
Published: May 2013

PROBLEM: ELISA is an ideal assay method for a large‐scale screening of anti‐sperm antibodies among a large number of infertile males. However, conventional ELISA with whole spermatozoa needs time‐consuming steps of centrifugation.

METHOD: A solid‐phase assay used for detecting anti‐sperm antibodies was established. This assay is suitable not only for detecting circulating anti‐sperm antibodies of IgG, IgM, and IgA subclass simultaneously but also for screening hybridomas secreting anti‐sperm monoclonal antibodies (mAbs). The microtiter plates, on which solubilized sperm antigens are fixed, can be stored at − 80°C for up to six months without losing reactivity with anti‐sperm antibodies.

RESULTS: Using this assay, 53 sera (13 were proven positive and 40 were proven negative for sperm agglutination antibody) were tested. Although the false‐negative rate was 0%, the false‐positive rate was 32%. One thousand one hundred sixty‐five supernatants from hybridomas constructed with splenocytes of mice who were hyperimmunized with human sperm and nonsecreting myeloma cells were tested by this solid‐phase assay and two anti‐sperm mAb secreting clones were selected and established.

CONCLUSIONS: It is recommended that for research work this assay could be used for the first screening of the hybridoma secreting anti‐sperm mAb, and for clinical use this assay might be suitable for the first screening of sera of infertile patients. However, conventional bioassays should follow to confirm the biological meaning of the positivity.
Cytokine‐Initiated Signal Transduction in Human Myometrial Cells
Authors: HERTELENDYM FRANK , ROMERO ROBERTO , MOLNÁR MIKLÓS , TODD HERBERT , BALDASSARE JOSEPH J.
Published: May 2013

PROBLEM: The objectives of this study were to evaluate interleukin‐1 (IL‐1) binding and some postreceptor actions of this cytokine and tumor necrosis factor (TNF) in human myometrial cells (HMC).

METHOD: Monolayer cultures of HMC were used to characterize binding and to measure cyclic (c)AMP, prostaglandin (PG)E2, and PGI2 production. Membrane preparations were used to assess ADP‐ribosylation and for immunoblotting.

RESULTS: HMC were found to specifically bind [125I]IL‐1 with an apparent Kd of 2 × 10−10M. Incubation of HMC with IL‐1 or TNF caused a time‐dependent and dose‐dependent accumulation of cAMP, as well as a significant potentiation of forskolin‐promoted cAMP production. These cytokines also increase PGE2 and PGI2 output, independently of the activation of adenylyl cyclase. IL‐1 treatment had no measurable effect either on cholera toxin‐mediated and pertussis toxin‐mediated ADP‐ribosylatidn, or on the amount of Gi proteins, as assessed by immunoblotting using a polyclonal antibody.

CONCLUSIONS: It is suggested that IL‐1 and TNF may activate one or more isoforms of the catalytic component of adenylyl cyclase, raising intracellular cAMP.
Hormonal Regulation of Complement Factor B in Human Endometrium
Authors: HASTY LISA A. , BROCKMAN WENDY W. , LAMBRIS JOHN D. , LYTTLE C. RICHARD
Published: May 2013

PROBLEM: Recent investigations have demonstrated the presence of complement components in human endometrium in a cycle‐specific manner. Luteal phase endometrium has been shown to synthesize complement C3 de novo, whereas proliferative endometrium produces little or no C3. Likewise factor B. which is critical to the activation of the alternative pathway of complement, has been shown to be present only in the glandular epithelium of luteal phase endometrium. This investigation was designed to determine if factor B is present in the endometrium in a high progesterone state such as pregnancy or with exogenous progesterone treatment.

METHOD: Endometrial biopsies were obtained from patients on progesterone therapy. The endometrium of early pregnancy was evaluated by obtaining biopsies from patients with ectopic pregnancies as well as from patients undergoing therapeutic termination. Immunohistochemistry was performed on each biopsy using monoclonal antibodies to factor B.

RESULTS: Our results demonstrate the presence of factor B in the glandular epithelial cells of the endometrium of patients treated with exogenous progesterone therapy. Additionally. factor B was localized to the glandular compartment of the endometrium from patients with ectopic gestations. Interestingly, the evaluation of an implantation site from an early gestation demonstrated factor B in the maternal decidua only; trophoblast did not exhibit the presence of factor B.

CONCLUSION: Factor B exists in the endometrium in a hormone‐dependent manner and is not expressed in fetal tissue in early gestation.
Association Between Maternal‐Fetal HLA‐DR Relationships and Fetal Growth
Authors: HOFF CHARLES , PEEVY KEITH J. , SPINNATO JOSEPH A. , GIATTINA KATHERINE , PETERSON RAYMOND D.A.
Published: May 2013

PROBLEM: To determine whether maternal‐fetal human leukocyte antigen (HLA) antigenic relationships are associated with differential fetal growth in weight.

METHOD: A cohort of 659 primigravid women were enrolled in this study in the prepartum period and their neonates were subsequently examined. Anthropometric, maternal cigarette smoking behavior, health, pregnancy, and delivery data were collected; serogenetic typing was conducted on maternal and cord bloods to determine maternal and neonatal HLA antigenic phenotypes. Women and their neonates were assigned to one of the four different types of maternal‐fetal relationships existing at each of the HLA‐A, B, DR, and DQ loci. Birthweights were treated quantitatively and qualitatively (neonates classified as growth‐retarded or normal).

RESULTS: After controlling for other factors influencing birthweight (e.g., smoking, maternal body size), significantly lower birthweight trends (P < .01) were found when neonates expressed a single HLA‐DR antigen and their mothers expressed a second HLA‐DR antigen that was foreign (allogeneic) to their neonate.

CONCLUSION: Our finding supports the hypothesis that lack of maternal immune exposure to fetal HLA antigens is associated with a slowing of fetal growth. However, in this situation slowed fetal growth is most likely to occur when the fetus is potentially exposed to maternal HLA‐DR alloantigens. We believe this sheds new light on immunologic events at the maternal‐fetal interface influencing fetal growth. We present one possible explanation to account for this finding.
Membrane‐Spanning Fcγ Receptor III Isoform Expressed on Human Placental Trophoblasts
Authors: NISHIKIORI NAOKO , KOYAMA MASAYASU , KIKUCHI TOMOYUKI , KIMURA TOSHIO , OZAKI MASAMI , HARADA SHIGENORI , SAJI FUMITAKA , TANIZAWA OSAMU
Published: May 2013

PROBLEM: Fc receptor for immunoglobulin (FcγR) is an important mediator of immunological functions in the feto‐maternal relationship. We have demonstrated by immunohistochemical means that three distinct classes of FcγRS are expressed in the different cell components of the human placenta.

METHOD: In this study, FcγRIII isoform expressed on placental trophoblasts (PTs) was investigated by indirect immunofluorescence and cDNA cloning. PTs, isolated from human term placenta by digestion with proteolytic enzyme, were reacted with monoclonal antibodies (MAb) against the Fc‐γRs and other surface markers of leukocytes and subjected to flow cytometric analysis.

RESULTS: PTs were positively stained with 3G8 and Leul lb against FcγRIII, partially stained with MAb against MHC class I, but not with 32.2 (FcγRI), IV3 (FcγRII), or MAbs against CD4, CD19, or CD56, indicating that only low affinity receptor, FC7RIII, is γexpressed on PTs. The DNA sequence of cloned FcγRIII CDNA from PTs by PCR was identical to that of natural killer (NK) cell isoform, including the position of the stop codon that differs from the granulocyte isoform by several nucleotide substitutions. We further analyzed the susceptibility of PTs against phosphatidylinositol specific phospholipase C (PI‐PLC) to determine the structural topology of PT isoform. While the reactivity with 3G8 on PTs was not influenced by treatment with PI‐PLC, that on granulocytes was significantly diminished with PI‐PLC.

CONCLUSIONS: This result confirmed that FcγRIII on PTs is a membrane‐spanning molecule, and that it is distinctive from PI anchoring FcγRIII on granulocytes.
Natural Killer Cell Activity of Lymphocytes Exposed to Ovine, Type I, Trophoblast Interferon
Authors: TUO WENBIN , OTT TROY L. , BAZER FULLER W.
Published: May 2013

PROBLEM: A member of newly described interferon (IFN) family, ovine trophoblast protein one (oTP‐1, now named as ovine type I trophoblast interferon, oTIFN), is a major secretory product of sheep conceptuses during days 11–21 of pregnancy. Type I trophoblast IFN shares high amino acid and nucleotide sequence homology with other mammalian IFN‐αs and has functional characteristics of IFNs such as antiproliferative, immunosuppressive, and antiviral activities. Moreover, oTIFN is responsible for the antiluteolytic signaling for early pregnancy recognition in sheep. It has been reported that natural killer (NK) cells are recruited to the pregnant uterus and contribute para‐immunotrophic effects on growth and differentiation of the conceptus (embryo/fetus and associated membranes). Because IFNs are potent modulators of NK cell activity, the present study examined interactions between oTIFN and sheep and pig NK cells.

METHODS: A 51Cr release assay was used to evaluate the effects of various concentrations of oTIFN and IFN‐α on sheep and pig NK cell activity against K562 and HeLaS3 cells in vitro. Dosages were tested at four effector to target cell ratios.

RESULTS: These data provide the first evidence that oTIFN is a potent NK cell activator in vitro. Recombinant human IFN‐α, (rhIFN‐α) and oTIFN increased (P < 0.01) ovine NK cell cytotoxicity for K562 cells at very low dosages (50 antiviral units (AVU)/ml). Lower doses of oTIFN (15 AVU/ml) were sufficient (P < 0.01) to enable sheep NK cells to kill ovine NK resistant cells, HeLa S3, in a dose‐dependent fashion. However, much higher concentrations (>400 AVU/ml) of oTIFN were needed to increase (P < 0.01) porcine NK cell cytotoxicity. This suggests that oTIFN has some species specificity in activating NK cells. The augmentation of NK cell activity by 100 AVU/ml of oTIFN was effectively neutralized by an anti‐oTIFN monoclonal antibody (HL‐98) at a dilution of 1:800.

CONCLUSIONS: Results of the present study clearly indicate that oTIFN is as active as rhIFN‐α, for activating NK cells in vitro. Immune cells, especially those recruited to the pregnant uterus by conceptus secretory products,−may be stimulated/activated by trophoblast IFNs. However, the function of activated NK cells during early pregnancy remains to be determined.
Is an Additional Vaccination Necessary for a Successful Second Pregnancy in Unexplained Recurrent Aborters Who Were Successfully Immunized With Their Husband's Lymphocytes Before the First Pregnancy?
Authors: TAKAKUWA KOICHI , HIGASHINO MASAHIKO , YASUDA MASAKO , ISHII SHIRO , UEDA HIROYUKI , ASANO KENSAKU , KAZAMA YOSHIKI , TANAKA KENICHI
Published: May 2013

PROBLEM: Is an additional immunotherapy necessary or not for patients who have obtained successful results after initial immunotherapy?

METHODS: The successive pregnancy outcome was analyzed in 22 patients out of 29 unexplained recurrent aborters who had undergone immunotherapy with their husband's lymphocytes according to our previously reported protocol (Takakuwa et al., Am J Reprod Immunol Microbiol. 10:1–9, 1986; Takakuwa et al., Am J Reprod Immunol. 23:37–41, 1990) and had obtained successful outcome between January 1983 and December 1989. In addition, the alteration of blocking antibodies (BAbs), which was evaluated by a one‐way mixed lymphocyte culture reaction (MLR) blocking assay between the spouses, was analyzed in 26 patients out of 29.

RESULTS: None of the 22 patients underwent further immunotherapy because a significantly high titer of MLR‐BAbs had been detected before the new pregnancy. In 19 out of the 21 patients (90.5%), pregnancy was successful.

CONCLUSION: Additional immunotherapy is not necessary for patients who have obtained successful results after the initial immunotherapy and are positive for MLR‐BAbs after their first delivery.