bluuebunny/crossref_metadata_2025 huggingface dataset:
Defense Proteins and Immune Complexes in the Blood Serum of Women With Inflammatory and Neoplastic Lesions of the Ovary
Authors: ZBROJA‐SONTAG WANDA
Published: May 2013

ABSTRACT: Blood sera of 181 women with inflammatory conditions and with benign and malignant neoplasms of the ovary and of 36 healthy women (control group) were examined by determining the levels of defense proteins: C‐reactive protein (CRP), alpha1 antitrypsin (A1AT), alpha2 macroglobulin (A2M), components C3 (beta1A‐globulin) and C4 (beta1E‐globulin) of the complement, immunoglobulins M (IgM), A (IgA), and G (IgG), as well as the concentration of circulating immune complexes (IC). In women with ovarian pathology, as compared with the control group, a significant increase of all the defense proteins was found (p < 0.05). The highest increase of the defense proteins, except in regard to A2M was observed in inflammation of the ovary. In malignant neoplasms of the ovary, the level of A2M was the highest and that of A1AT as high as in inflammatory conditions. There was also a high percentage of increased CRP values (57%). The concentration of the remaining proteins under study, although significantly higher than in the control group (p < 0.05), was in this group the lowest in comparison with the groups of ovarian inflammation and benign neoplasms, especially with regard to component C3 of the complement and to IgM and IgG (p < 0.05). Significantly increased values of IC (p < 0.05) were observed in ovarian inflammation (73.2%) and in benign neoplasms (25.0%). In the group of malignant neoplasms (15.4%) and in healthy women (5.6%), the increased IC values were not statistically significant (p > 0.05).
Blocking Effects of Maternal Serum‐IgG and Placental Eluate‐IgG on Materno‐Fetal Mixed Lymphocyte Reaction and Their Individual Specificity*
Authors: KAJINO TORU , KANAZAWA KOJI , TAKEUCHI SHOSHICHI
Published: May 2013

ABSTRACT: The purpose of this study is to clarify the problem regarding individual specificity of maternal serum immunoglobulin G (IgG) and placental eluate IgG that block the unidirectional mixed lymphocyte reaction (MLR) in which maternal and cord blood lymphocytes (lys) were used as responder and stimulator, respectively. Both maternal serum IgG and placental eluate IgG were found to include blocking antibodies (BA) which showed similar blocking effect on MLR. But, in the case of serum IgG, individual specificity of BA on MLR was observed to be less great in MLR done using whole lys or T‐enriched lys of the cord blood as stimulator, whereas it became greatly significant when MLR was done using B‐enriched lys of the cord blood as stimulator. This was the case even in MLR done by using unrelated maternal lys as responder. On the other hand, in the case of placental eluate, it was found to be greater when compared to serum IgG even where MLR was done using not only B lys but also T lys as stimulator. It is thus strongly suggested that BA include individual specific antibodies against fetal B lys having HLA‐D/DR locus determinants in addition to individually nonspecific antibodies common to pregnancy which may be produced to a “yet unknown fetal antigen.”
Demonstration That Milk Cells Invade the Suckling Neonatal Mouse*
Authors: WEILER IVAN JEANNE , HICKLER WOLFGANG , SPRENGER RENATE
Published: May 2013

ABSTRACT: Mouse milk cells were stained with rhodamine or fluorescein isothiocyanate and fed to young suckling mice. By visual examination of serial sections and by flow cytofluorometry, we were able to demonstrate directly the presence of these cells in peripheral tissues.

It was estimated that at least 0.1% of the fed cells might infiltrate the young mouse, which is initially immunologically defenseless. This is in accordance with evidence from many sources for activity of maternally‐derived lymphoid cells in young rodents.
Monoclonal Antibody Inducing Human Sperm Agglutination
Authors: YAN YUAN CHANG , WANG LIN FANG , SATO EIMEI , KOIDE S.S.
Published: May 2013

ABSTRACT: A mouse fused splenic cell line producing antibody that induces agglutination of human sperm was cultivated. The antibody was localized to the midpiece and tail of human spermatozoa by an indirect immunofluorescence method. The interacting antigen was identified in a sperm membrane protein preparation by SDS gel/protein blot immunofluorescence method. The molecular weight of the antigen was in the vicinity of 84 Kdaltons.
Induction of High Titer Mouse‐Antihuman Spermatozoal Antibodies by Liposome Incorporation of Spermatozoal Membrane Antigens
Authors: METTLER L. , CZUPPON A.B. , BUCHHEIM W. , BAUKLOH V. , GHYCZY M. , ETSCHENBERG J. , HOLSTEIN A.F.
Published: May 2013

ABSTRACT: This is the first report of induction of high titer mouse‐antihuman spermatozoal antibodies using a spermatozoal antigen incorporated in liposomes. The peptide antigens that were incorporated into pure phosphatidylcholine liposomes are known to react with the naturally occurring antibodies in sera of sterile patients. Electron microscopy and the complement‐dependent glucose release tests indicated that the peptides were entrapped in the aqueous phase between the lipid bilayers in the liposome vehicles. This is in contrast to most other liposomal antigens that are expressed on the outer lipid surface. In view of the nontoxic nature of the liposomes these results could be of some interest in immunological fertility regulation trials with appropriate antigens in an allogeneic system.
Induction of Suppressor Cell Activity by Human Pregnancy Serum
Authors: NAKAMURA TSUNEYOSHI , PERSELLIN ROBERT H. , RUSSELL I. JON
Published: May 2013

ABSTRACT: Third trimester pregnancy serum caused a dose‐dependent inhibition of the one‐way allogeneic mixed leukocyte reaction (MLR) through an effect that occurred during the first 48 hours of culture. Pregnancy serum also inhibited the mitogenic responsiveness of normal mononuclear leukocytes to concanavalin A (Con A) while the responses to phytohemagglutinin (PHA) and pokeweed mitogen (PWM) were less affected. Preincubation of normal peripheral blood mononuclear cells (PBMC) for 48 hours with 10% pregnancy serum enhanced a suppressor activity transferable with cells. These pregnancy serum‐induced effector cells suppressed the MLR only when they were autologous to the responder population (p < 0.05). The same suppressor cell preparation inhibited the proliferative responses of autologous PBMC to Con A (p < 0.001) and PWM (p < 0.05). These data suggest that one or more factors in pregnancy serum can induce or enhance suppressor cell activity in vitro. A comparable increase in suppressor cell activity in vivo may be responsible for blocking maternal rejection of the fetus and for the observed improvement in clinical activity of rheumatoid arthritis during pregnancy.
Mononuclear Cell Subpopulations During Normal Pregnancy: I. Analysis of Cell Surface Markers Using Conventional Techniques and Monoclonal Antibodies
Authors: LUCIVERO G. , SELVAGGI L. , DELL'OSSO A. , ANTONACI S. , IANNONE A. , BETTOCCHI S. , BONOMO L.
Published: May 2013

ABSTRACT: Previous reports indicate depression of cellular immunity during pregnancy. In order to get a deeper insight into the immunoregulatory subsets of mononuclear cells in pregnant women, we analyzed the percentages of T and B lymphocytes, T cell subsets and cells expressing IgG‐Fc and C3 receptors, and M1‐ and Ia‐like antigens in venous blood samples from healthy women and pregnant women at different gestational ages and after delivery. Even if a trend toward a progressive decrease of T lymphocytes and T4+ helper subset was observed throughout pregnancy, along with a slight increase in the percentages of cells bearing C3 receptors, M1, and Ia‐like antigens in the second trimester of gestation, analysis of variance of our data did not demonstrate statistical significance in the observed changes. Therefore we conclude that the pregnancy‐induced suppression of maternal responsiveness towards alloantigens and/or mitogens is not reflected by significant changes in the distribution of circulating mononuclear cells and immunoregulatory T‐Iymphocyte subsets as defined by the currently available surface marker techniques.
Cell‐Mediated Immunity in the Blood of Women With Inflammatory and Neoplastic Lesions of the Ovary
Authors: ZBROJA‐SONTAG WANDA
Published: May 2013

ABSTRACT: Blood samples from 181 women with inflammatory conditions and with benign or malignant neoplasms of the ovary and from 36 healthy women (control group) were examined by determining the following indicators of cell‐mediated immunity: leukocyte count, lymphocyte count, absolute number of lymphocytes, percentage of rosette‐forming T lymphocytes, absolute number of T lymphocytes, and phagocytic activity of granulocytes. In women with ovary inflammation, there was a very high phagocytic activity of granulocytes. It correlated with an increased leukocyte count. The activity and number of T lymphocytes were very low in this group. In this group was also the lowest percentage of lymphocytes. In the group with malignancies of the ovary, there were the same very low activity and number of T lymphocytes. The values of T lymphocytes in inflammations and malignancies were almost equal and did not differ significantly from one another. The phagocytic activity of granulocytes was the lowest in women with malignancies of the ovary. In blood of women with benign neoplasms of the ovary, the activity and number of T lymphocytes and the activity of granulocytes were significantly higher than in the malignancies group.
Habitual Abortion: Parental Sharing of HLA Antigens, Absence of Maternal Blocking Antibody, and Suppression of Maternal Lymphocytes
Authors: UNANDER A. MARGARETA , OLDING LARS B.
Published: May 2013

ABSTRACT: The immunologic responsiveness of eight women who habitually abort has been investigated. All shared an HLA‐A or B antigen with their husbands. Sharing of an HLA‐DR antigen was found in seven couples, one of which also had a second DR antigen in common. The probability for this high frequency of HLA‐DR sharing is negligible (p = 0.0004), as calculated from the antigen frequencies among Europeans. Cells from the woman with two shared DR antigens displayed a minor response to her husband's cells but reacted strongly to control cells, whereas the other women's cells reacted normally to cells from both their husbands and controls in one‐way mixed lymphocyte culture (MLC). Only minor cytotoxicity was displayed by women's cells in a direct cell‐mediated lympholysis (CML) assay, but they mounted normal cytotoxic responses against both husbands' cells and control cells in an amplified CML assay. The sera from six of the habitually aborting women displayed no blocking activity in one‐way MLC, and seven of them had no cytotoxic antibodies. Cells from all habitual aborters were suppressed in two‐way MLC by cells from husbands and most controls. We hypothesize that increases in HLA compatibility between mother and fetus and in maternal susceptibility to suppressive influences are in some way linked to a deficiency in the development of anti‐fetal antibody during pregnancy. As a consequence, the fetus may be deprived of the protection by maternal blocking antibody, which may allow maternal cytotoxic reactions to cause abortion.
Immunological Response of Guinea Pigs Sensitized With Homologous Antigens
Authors: ALONSO ANGEL , DE CEREZO JOSEFINA M.S. , GONZALEZ NÉLIDA , BUENO MARÍA P. , MANCINI ROBERTO S.
Published: May 2013

ABSTRACT: The chemical composition of some “immunologically” pure antigens isolated from guinea pig testis and spermatozoa was correlated with their antigenic behavior. Their immunological responses were compared to select the best materials for a further isolation of chemically pure antigens. The glycoprotein extracted from the spermatozoa (T Gly) has the highest immunological potency and seems to be a T and B, depending antigen, able to induce high humoral and cell responses producing germinal cell damage, testicular lesions, and aspermatogenesis.
Reproductive Immunology News*
Authors: Gleicher Norbert
Published: May 2013

The NICHD once again has provided the AJRI with grant summaries of projects in reproductive immunology awarded grant funding for fiscal year 1983. In each issue of Volume 6, we will feature descriptions of some of those grants. The information given in each summary was provided in the grant applicaiton by the principal investigator.

The Editorial Board of the AJRI would like to thank the NICHD for its cooperation in regard to this feature of the Reproductive Immunology News section of the Journal It is their hope that continuing to publish this information will continue to facilitate increased communication among investigators in the field of reproductive immunology.
Characterization of Antibodies to a Placenta‐Specific Antigen Cross‐Reacting With a Choriocarcinoma Cell Line
Authors: Sinha Subrata , Bhattacharya Alok , Band Abdul Hamid , Gaur Amitabh , Strivastava L.M. , Talwar G.P.
Published: May 2013

ABSTRACT: Antibodies were raised in rabbits to purified human term placental villous plasma membrane. These were cytotoxic to human peripheral blood lymphocytes and manifested cross‐reactivity to kidney and liver. After absorption with these tissues, reactivity was retained with placental villous plasma membrane. The placental‐specific antiserum was fully reactive with BeWo choriocarcinoma cells, but was devoid of reaction with a human myeloma tumor cell line (GM2132). The antibodies were cytotoxic to BeWo in the presence of complement, as determined by trypan blue uptake, 51Cr release, and inhibition of uptake of 3H‐thymidine.
Transplacental Alloimmunization to Fetal Erythrocytes in Rhesus Monkeys (Macaca mulatta)
Authors: Stong Robin C. , Nelson Kathleen H. , Stone W.H.
Published: May 2013

ABSTRACT: Evidence of transplacental immunization in rhesus monkeys was obtained by testing postpartum sera from 252 females for antierythrocyte agglutinins. One‐third of the sera contained antibodies reactive with the mates' or the newborns' cells. Fetal erythrocytes were detected in the maternal circulation as early as 8 weeks after conception and as late as 12 days postpartum. Maternal antibodies were detected as early as 2 months after conception and persisted for more than 3 weeks postpartum. It was concluded that the fetal cells stimulated antibody production. Several features of transplacental immunization differ between rhesus monkeys and humans. A parity effect was not observed in rhesus. In fact, 33% of primiparous rhesus females produced antibodies. Also, several of the different allogeneic blood group factors appeared to be immunogenic but differed in immunopotency. Finally, direct antiglobulin tests indicated that erythrocytes of 11% of newborns were coated with maternal antibodies. Nevertheless, in contrast to humans, hemolytic disease was not observed.
Maternal Antipaternal Immunity in Couples Predisposed to Repeated Pregnancy Losses
Authors: McCONNACHIE PETER R. , McINTYRE JOHN A.
Published: May 2013

ABSTRACT: Maternal antipaternal immunity was compared between a group of 12 couples with normal reproductive histories and a group of 13 couples with obstetrical histories of more than one consecutive stillbirth or repeated pregnancy loss subsequent to having had successfully carried to term. Couples were studied for maternal‐paternal HLA antigen sharing and for maternal antihusband immunity in tests detecting both complement dependent and independent antibodies as well as in an assay measuring direct cell‐mediated cytotoxicity. There was no significant difference in the proportion of HLA antigen sharing between these two groups; however, the women in the pregnancy loss group demonstrated significantly more antipaternal immunity than their normal childbearing counterparts and more than previously described for aborting couples who share HLA antigens. Screening of the cytotoxic sera on a well characterized cell panel failed to reveal alloreactivity patterns consistent with paternal HLA antigen profiles. These data demonstrate that certain women who suffer recurrent pregnancy losses can mount vigorous immune responses to paternal lymphocyte antigens. We propose that the appearance of such lymphocytotoxins represents inappropriate maternal immune responses to fetal extraembryonic antigens that may subsequently result in fetal demise
Antibodies Against the β‐Subunit of Ovine Luteinizing Hormone Can Decrease the Clearance of Human Chorionic Gonadotropin in Rhesus Monkeys*
Authors: YAMAMOTO YUKIO , GUNSALUS GLEN L. , SUNDARAM KALYAN , THAU ROSEMARIE B.
Published: May 2013

ABSTRACT: Contraceptive vaccines based on active immunization against gonadotropic hormones are being investigated in humans and other primates. Immunization against the β‐subunit of ovine luteinizing hormone (oLHβ) reduces fertility in rhesus monkeys by inducing inadequate luteal phases and preventing corpus luteum rescue by rhesus chorionic gonadotropin (rhCG). These effects result from the cross‐reactions of the oLHβ‐antibodies with rhCG and rhLH. We used human CG (hCG), which also cross‐reacts strongly with anti‐oLHβ to examine how the circulating oLHβ‐antibodies affect the metabolic clearance rates (MCR) of hCG in rhesus monkeys. 125I‐hCG was injected into four nonimmunized and seven immunized monkeys and blood was collected at frequent intervals over 7 days. Total and immunoprecipitable radioactivity did not differ significantly, suggesting that the radioactivity in the plasma consisted almost entirely of 125I‐hCG. This was confirmed by column chromatography. The MCR (mean ± SE) was significantly lower (p < 0.001) in six immunized monkeys (0.35 ± 0.06 liters/day) as compared to controls (1.19 ± 0.09 liters/day). The hCG disappearance curve in control monkeys was best described by a two‐compartmental system (slow and fast) while an additional third (intermediate) compartment of distribution was typical for immunized animals. The half‐lives of hCG for the two exponentials corresponding to the slow and fast components of distribution were not significantly different between the two groups. One immunized monkey had a MCR (1.44 liters/day) that was much greater than the MCR of the other six. This monkey cleared a significantly smaller proportion of hCG in the slow and a higher proportion in the intermediate compartment and unlike the others, formed a circulating immune complex capable of binding hCG that was significantly larger than the antibody‐hCG complexes found in the other six immunized animals. We conclude that circulating antibodies to oLHβ reduced the MCR of hCG in six of seven monkeys. The decreased MCR found in immunized monkeys is associated with a shift in clearance from the “fast” to the “slow” compartment as well as the addition of an intermediate compartment of distribution. Plasma disappearance rates of hCG depend on the size of the antibody hCG complex.
Sperm Antibodies, Immunoglobulins, and Complement in Human Follicular Fluid
Authors: CLARKE G.N. , HSIEH C. , KOH S.H. , CAUCHI M.N.
Published: May 2013

ABSTRACT: The levels of immunoglobulins (IgG, IgA, IgM), complement (C3, C4, C1EI) and sperm antibodies were determined in plasma and follicular fluid samples from 26 patients undergoing in vitro fertilization (IVF) treatment. The results show that IgG, IgA, C3, C4, and C1EI concentrations in follicular fluid are similar to plasma concentrations (63.1–96.1% of plasma levels). The follicular fluid concentration of IgM was severely reduced, however, being only approximately 10% of plasma concentrations. Sperm antibody titres were compared in three patients using sperm agglutination, immobilization, and immunobead binding. The titres in plasma and follicular fluid were similar, apart from antibodies of IgM class, which were undetectable in follicular fluid.
Immune Monitoring of Pregnancy in Renal Transplanted Patients
Authors: BIRKELAND S.A. , KRISTOFFERSEN K.
Published: May 2013

ABSTRACT: Immune studies were performed in eight pregnancies occurring after renal transplantation (tx). In three cases a regular, long‐term immune monitoring before, during, and after the pregnancy was possible. The results showed 1) no changes concerning OKT 3, 4, 8, and SMIg‐defined T‐ and B‐lymphocyte subsets in the mothers: 2) a higher level of spontaneous E‐ and normal E‐rosette‐defined T‐subsets in the mothers rejecting their allografts; 3) a stronger response to mitogen and antigen stimulation and a stronger mixed lymphocyte culture (MLC) response towards the infants also in those rejecting their grafts; 4) the offspring of the transplanted mothers showed a weaker immune responder state compared with the offspring of six age‐matched normal mothers.
Local Active Suppression by Suppressor Cells in the Decidua: A Review
Authors: CLARK DAVID A. , SLAPSYS RENATA , CROY B. ANNE , KRCEK JERRY , ROSSANT JANET
Published: May 2013

ABSTRACT: The immunological survival of the antigen‐bearing mammalian feto‐placental unit is determined by the functional properties of the tissues at the feto‐maternal interface. Antigen‐specific systemic suppressor mechanisms such as suppressor T cells and non‐antigen‐specific suppressive serum factors appear not to play a major role in protection of the fetus. A novel type of non‐MHC specific suppressor cell accumulates locally in the decidua of successfully allopregnant mice. This decidua‐associated suppressor is a small lymphocytic cell possessing cytoplasmic granules, lacks T cell markers, and is deficient in number and activity at the implantation sites of viable xenogeneic Mus caroli embryos gestating in the uterus of Mus musculus animals at the time that maternal lymphoid cells begin to infiltrate the xenoembryos. These Mus caroli embryos subsequently resorb. Further experimental studies suggest that the trophoblast cells associated with successful pregnancy recruit bone‐marrow derived maternal non‐T suppressor cells to the decidua and thus, by an indirect mechanism, may act to protect the fetus from effector cells of the mother's immune system.
Monoclonal Antibody to Human Cytotrophoblast
Authors: LOKE Y.W. , DAY SUSAN
Published: May 2013

ABSTRACT: A monoclonal antibody (18B/A5) has been generated against human first trimester trophoblast membranes which, unlike others so far reported in the literature, reacted only with cytotrophoblast and not with syncytiotrophoblast. Although the identity of the target antigen has not yet been established, the antibody could be a valuable tool for the identification and eventual separation and purification of human cytotrophoblast cells.
Comparison of Corynebacterium Parvum With Freund's Complete Adjuvant as a Potentiator of the Immune Response To β‐Human Chorionic Gonadotropin Linked to Tetanus Toxoid
Authors: COVEY DANA C. , LAURENCE KENNETH A.
Published: May 2013

ABSTRACT: Corynebacterium parvum was compared with Freund's complete adjuvant (FCA) for potentiation of the rabbit immune response to β‐human chorionic gonadotropin linked to tetanus toxoid (β‐hCG‐TT). With each adjuvant, antibodies to hCG were detected using passive hemagglutination. Higher antibody titers were produced by FCA‐treated animals. The ability of antisera to β‐hCG‐TT to neutralize the biological action of native hCG was determined by the rat uterine weight assay. Anti‐β‐hCG‐TT sera from C. parvum‐treated rabbits were not significantly different (p > 0.10) from FCA potentiated anti‐β‐hCG‐TT sera in neutralizing hCG‐induced uterine weight gain. C. parvum has potential for future application in active immunization studies of fertility regulation.
Decline in Sialic Acid Composition of Cellular Membranes Isolated From Ovine Corpora Lutea During Prostaglandin‐Induced Luteolysis: Apparent Independence of Autoimmune Recognition
Authors: STEADMAN L.E. , JOHNSON G.A. , BELDEN E.L. , MURDOCH W.J.
Published: May 2013

ABSTRACT: Sialic acid was quantified in plasma membranes of corpora lutea isolated during prostaglandin (PG) F2α‐induced luteolysis in sheep. Concentrations of sialic acid within membranes decreased after injection of PGF2α, and before signs of luteal regression (i.e., a decline in tissue concentrations of progesterone) were manifested. Removal of residues of sialic acid from luteal membranes was not associated with cellular binding of gamma globulin, as monitored by indirect immunofluorescence microscopy. We suggest that desialylation of luteal membranes could be an important aspect of the mechanism of luteolysis. Such a process does not appear to involve participation of autoantibody.
Immunological Studies of Trophoblast Antigens: No Evidence for Human Leucocyte Antigen (HLA) Linkage
Authors: MacLEOD A.M. , STEWART K.N. , CATTO G.R.D. , MCINTYRE J.A.
Published: May 2013

ABSTRACT: Parental disparity for trophoblast‐lymphocyte crossreactive (TLX) antigens may promote successful pregnancy. A TLX antigen system has been defined on peripheral blood lymphocytes by heteroantisera. More recently, we have reported additional activity against antigens on B lymphocytes alone termed trophoblast‐B lymphocyte crossreactive (TBX) antigens. In the present study we have investigated ten TLX sera in order to determine if their target antigens are linked to the human leucocyte antigen (HLA) gene complex. The sera showed no selective activity when tested against target B lymphocytes from ten normal donors. Cytotoxic activity of TLX antisera against peripheral blood lymphocytes from six normal donors was not reduced when the class I HLA antigens of the target cells were blocked with a monoclonal antibody (PA 2.6). Similarly the cytotoxic activity of both TBX antisera against B lymphocytes from six normal donors was not decreased when class II HLA antigens were blocked by a monoclonal antibody (FMC 4). Within a family the cytotoxic activity of the TLX antisera was absorbed equally by lymphocytes from siblings who shared neither HLA haplotype. Antibody content in TLX and TBX antisera is not directed toward the classically defined HLA class I or class II antigens and is not linked to the HLA gene complex.
In Vitro Sensitivity of Candida (Torulopsis) glabrata to Clotrimazole
Authors: FAGNANT JOAN E. , CLARK RICHARD B. , MONIF GILLES R.G.
Published: May 2013

ABSTRACT: Vulvovaginitis caused by Candida (Torulopsis) glabrata is often refractory to intravaginal imidazole therapy. Clotrimazole achieves its fungistatic activity for Candida albicans and C. glabrata by inhibiting different steps in intermediary cell metabolism. For C. glabrata, alkylation precedes dimethylation. The possibility that this altered sequence might account for the relative therapeutic nonresponsiveness was studied by determining comparative minimal inhibitory concentrations (MICs) of clotrimazole. In vitro analyses of ten strains of C. glabrata and 30 control strains of C. albicans performed using both agar and broth dilution tests revealed that fourfold lower MICs were consistently demonstrable with C. glabrata, irrespective of inoculum size. The data suggest that clinical difficulties encountered in the therapy of torulopsis vulvovaginitis probably represent the inability of intravaginal medication to eradicate urethral/urinary bladder colonization and subsequent reinfection rather than true therapeutic failures.
In Vitro and In Vivo Embryo Toxicity of Antilaminin Antibodies in the Rat
Authors: WEEKS BENJAMIN S. , KLEIN NORMAN W.
Published: May 2013

ABSTRACT: Rats mated after laminin immunization had higher frequencies of resorptions (57%) than those immunized with bovine serum albumin (20%) and had sera that were toxic to cultured rat embryos. In addition, sera from rats immunized with laminin A chains but not B chains were toxic to cultured embryos. The toxicity of sera to embryos was related to the reactivity of sera to specific laminin fragments rather than to sera IgG levels against intact laminin. In addition, resorptions in pregnant rats immunized with laminin were not related to the sera antilaminin IgG levels. However, levels of uterine antilaminin IgA, the predominant uterine isotype, increased considerably during the first 3.5 days of pregnancy, while sera antilaminin IgG remained constant.
Suppression of Lymphocyte Activation by a Soluble Factor Released From the Human Placental Chorionic Membrane: Chemical Analysis and Functional Characterization
Authors: SKIBIN AGNETA , QUASTEL MICHAEL R. , KUPERMAN ODED , SEGAL SHRAGA
Published: May 2013

ABSTRACT: A heat‐resistant factor that markedly and reversibly inhibits human lymphocyte activation was discovered in culture medium of human placental chorion. The chorionic factor inhibits lymphocyte proliferation in response to polyclonal mitogens and in the mixed leukocyte response. The inhibitory action is most effective if the factor is added during the first 24 h of lymphocyte culture and is reversible. The chorionic factor is sensitive to proteinase K, pepsin and bovine pancreatic protease; its activity is completely lost after papain digestion and following treatment with trichloracetic acid (TCA). The factor prevents the expression of IL‐2 receptors and class II MHC glycoproteins (HLA‐DR) on phytohemagglutinin‐stimulated PBMC but does not affect the expression of MHC class I molecules. It inhibits the replication of IL‐2‐dependent CTLD cells but is without effect on the growth of various human or murine cell lines or acute leukemic cells. Human placental chorion is thus capable of releasing in vitro a nontoxic heat‐resistant factor with protein characteristics that reversibly inhibits processes associated with the early stages of lymphoid cell triggering. This factor may play a role in an immunoregulatory mechanism that prevents maternally mediated immune rejection of the conceptus.
Membrane Fluidity of Trophoblast Cells and Susceptibility to Natural Cytotoxicity
Authors: SZEKERES‐BARTHO JULIA , NÉMETH A. , VARGA P. , CSERNUS V. , KÖSZEGI T. , PAÁL M.
Published: May 2013

ABSTRACT: This study examines the relationship between membrane lipid microviscosity and susceptibility of villous trophoblast to lysis by natural cytotoxic cells.

Trophoblast‐enriched cell suspensions prepared from term human placentae were treated with cholesteryl hemisuccinate (CHS)—a modulator of membrane lipid microviscosity. CHS‐treated cells were more susceptible targets for natural lymphocyte cytotoxicity than were untreated controls. In binding experiments, increased binding of lymphocytes to CHS‐treated target cells was found. Preincubation with progesterone prevented membrane rigidification by CHS. Progesterone, Cortisol, and estriol restored the impaired resistance of CHS‐treated trophoblast cells to lysis.

We determined microviscosity and progesterone concentration in villous surface membranes, prepared from placentae from idiopathic spontaneous abortions and normal first‐trimester pregnancies.

An inverse relationship was found between progesterone content and microviscosity of the membranes. Microviscosity of the membranes from abortion placentae was significantly higher (P < .01) and progesterone concentration was significantly lower (P < .001) than those in the membranes of normal first trimester placentae.
Inhibition of Lymphocyte Proliferation by Ovine Trophoblast Protein‐1 and a High Molecular Weight Glycoprotein Produced by the Peri‐Implantation Sheep Conceptus
Authors: NEWTON G.R. , VALLET J.L. , HANSEN P.J. , BAZER F.W.
Published: May 2013

ABSTRACT: Sheep conceptuses were flushed from uteri on day 16 of pregnancy and cultured in vitro. Three peaks of immunosuppressive activity (i.e., ability to inhibit [3H]thymidine incorporation into mitogen‐stimulated lymphocytes) were localized in conceptus‐conditioned culture medium: one corresponding to a high‐molecular‐weight glycoprotein (HMWG; Mr = 800–900 kDa), one corresponding to ovine trophoblast protein‐1 (oTP‐1), the antiluteolytic, interferon‐like molecule of the sheep conceptus, and a third fraction containing a previously undescribed molecule with a molecular weight between 10 and 14 kDa. Both HMWG and oTP‐1 inhibited lymphocyte proliferation in a dose‐dependent manner. Delaying the addition of HMWG or oTP‐1 until 24 h after the addition of phytohemagglutinin (PHA) reduced the degree of suppression relative to cultures where HMWG and oTP‐1 were added at the time of PHA addition. Nonetheless, the molecules were still capable of inhibiting proliferation. Addition of human recombinant IL‐2 (0–65 U/ml) to PHA‐stimulated lymphocyte cultures could not reverse HMWG‐ or oTP‐1‐induced suppression of [3H]thymidine uptake by PHA‐stimulated cells. Furthermore, treatment of lymphocytes with HMWG or oTP‐1 suppressed IL‐2‐induced proliferation. Therefore, HMWG and oTP‐1 affect both early and later events in the in vitro proliferative response of mitogen‐stimulated lymphocytes, including responsiveness to IL‐2.
Fetus as an Allograft: Noncytotoxic Maternal Antibodies to HLA‐Linked Paternal Antigens
Authors: INNES A. , CUNNINGHAM C. , POWER D.A. , CATTO G.R.D.
Published: May 2013

ABSTRACT:A cellular enzyme‐linked immunospecific assay (CELISA) was used to monitor maternal humoral responses in human pregnancy. Noncytotoxic IgG antibodies to paternal lymphocytes were detected in sera from 6 of 20 normal first trimester primigravidae and 6 of 13 multiparae. No antibody activity against lymphocytes from their partners was detected in sera from any of the 15 nulliparous women. The differences in antibody response between primigravidae and nulliparae (P= 0.024) and between multiparae and nulliparae (P = 0.005) were statistically significant. Lymphocytotoic antibodies to T‐ and B‐lym‐phocytes were present in sera from three multiparae, but from none of the women in the other two groups. Family studies indicated that the noncytotoxic pregnancy‐associated maternal antibodies were directed to HLA‐linked antigens (P< 0.001). Evidence obtained using cell panels and platelet absorption suggested, however, that these antibodies were not directed to the currently recognized HLA specificities (HLA‐A, ‐B, ‐C, or ‐DR).
Attempts to Identify Trophoblast in Peripheral Blood During Pregnancy Using Monoclonal Antibody H315
Authors: POOL CLAIRE , APLIN JOHN D. , TAYLOR G. MALCOLM , BOYD ROBERT D.H. , DONNAI PAUL
Published: May 2013

ABSTRACT: In view of its potential importance in antenatal diagnosis of genetic disease, we have used flow cytometry to attempt to enumerate trophoblast cells in the peripheral blood of pregnant females. As a trophoblast marker we have used the monoclonal antibody (MAB) H315, which reacts with placental‐type alkaline phosphatase (PLAP) associated with the cell‐membrane of the syncytiotrophoblast. We analyzed blood leucocytes from 62 pregnant females between 6 and 22 weeks' gestation, 26 nonpregnant females, and 21 males. Overall, we detected more H315+ cells in pregnant than in nonpregnant female blood but, because of the wide variation in both groups, we found no statistically significant difference between them. We were unable to detect H315+ cells in 25% of pregnant females within the resolution of the technique. There was, in contrast, a significant difference between the number of H315 + cells in pregnant females and males. Because we were unable to demonstrate other trophoblast markers on H315+ cells isolated from maternal peripheral blood, the origin of these cells is unclear. This implies that these cells may be an unsuitable source of material for antenatal diagnosis.
Antigenicity of Basement Membrane Fractions Obtained From Rat Seminiferous Tubules
Authors: ARECES LILIANA B. , BONINO MIRTHA BISCOGLIO DE JIMENEZ , CASCONE OSVALDO , PUIG RAÚL , DENDUCHIS BERTA
Published: May 2013

ABSTRACT: A noncollagenous fraction of basement membrane (D‐STBM) obtained from rat testes was submitted to sodium dodecyl sulfate‐polyacrylamide slab gel electrophoresis (SDS‐PAGE), and eight well‐defined bands were detected. A cross‐reaction with an antiserum against laminin was revealed by immunoblotting in five bands, with molecular weights ranging from 54 to 64 kDa. No further resolution of these components could be obtained by size exclusion and ionic exchange chromatography.

Fifty‐two percent of the rats immunized with DSTBM and adjuvants developed a mild multifocal damage of the testis. The lesions were characterized by foci of seminiferous tubules with different degrees of sloughing and/or atrophy of the germinal epithelium. Giant multinucleated cells were frequently seen, and mild interstitial mononuclear cell infiltrates were also detected. By immunofluorescence, deposits of rat IgG with a faint discontinuous linear pattern were observed along the walls of the seminiferous tubules. Circulating antibodies to D‐STBM were detected by ELISA in 100% of the rats, whereas in a cross‐reaction with laminin antibodies were detected in only 63%. All rats studied revealed a positive delayed type of hypersensitivity (DTH) response to D‐STBM.

None of the control rats injected with saline and adjuvants presented circulating antibodies to D‐STBM or laminin or a positive DTH reaction to D‐STBM. Some control group rats (10%) revealed few isolated seminiferous tubules with some degree of sloughing of the germinal epithelium.
An Evaluation of the Accuracy of Screening Antisperm Antibodies Using the Combined GAM Immunobead
Authors: VILLINES PATRICIA M. , KINCADE RORY S. , COULAM CAROLYN B. , CRITSER ELIZABETH S. , CRITSER JOHN K.
Published: May 2013

ABSTRACT: A simplified method of screening for antisperm antibodies has been previously described using rabbit antihuman immunoglobulin immunobeads (GAM beads). These reports have indicated a high correlation between the GAM bead and maximal individual isotype binding. However, preliminary data in our laboratory (using a 14% cut‐off criterion) indicated a high frequency of samples with positive GAM bead, but negative individual isotype results. This study was conducted to evaluate more critically the use of the GAM bead for initial antisperm antibody screening. Immunobead binding tests were performed on 98 undiluted sera. The maximal binding of the individual isotype beads (IgG, IgA, or IgM) and the GAM beads were significantly correlated (r = 0.94, P = 0.0001). However, when results were categorized as positive or negative, there was a significantly lower frequency (P < 0.05) of positive samples using the individual isotype (46.9%) than using the GAM approach (58.2%). These data support the hypothesis that, based on continuous percent binding, the GAM bead method produces results similar to individual isotype testing. However, when data are interpreted categorically, the results may differ significantly.
Prostacyclin Release and Cytotoxicity of Peritoneal Cells Are Inversely Related in Pregnant and Non‐Pregnant Mice Infected With Herpes Simplex Virus
Authors: LOFTIN KARIN C. , GONIK BERNARD , WALSH SCOTT W. , KUMARAN PRIYA , RAMIREZ CHRISTINA
Published: May 2013

ABSTRACT: Cytotoxicity of peritoneal cells in a HSV‐infected murine model is attenuated in late pregnancy. Prostacyclin (PGI2) is elevated at this time in reproductive tissues and has been implicated in the regulation of the immune response. The purpose of this study was to estimate PGI2 in the peritoneal wash or culture supernatants of peritoneal cells obtained from uninfected and HSV‐infected pregnant and virgin mice using a radioimmunoassay for 6‐keto‐prostaglandin F1α. The peritoneal wash of uninfected pregnant and virgin mice contained high levels of 6‐keto‐PGF1α, 505 ± 51 pg/100 μ1, (mean ± S.E., n = 15), and 200 ± 19 pg/100 μ1, (n = 30), ad did peritoneal effector and target cell cultures (1,159 ± 118 pg/100 μ1, n = 6, and 1,057 ± 207 pg/100 μ1, n = 7), respectively. HSV‐infection induced in vitro cytotoxicity and suppressed the release of 6‐keto‐PGF1α (r = −0.897, P < 0.05, n = 18). Its concentration was significantly higher (14‐fold, P < .05) in the peritoneal wash, but not in the cell culture, of pregnant (212 ± 29 pg/100 μ1, n = 19) as compared to virgin mice (18.5 ± 3.4 pg/100 μ1, n = 27). The levels of 6‐keto‐PGF1α were inversely correlated (P < .05) with the combined effects of HSV‐infection and cytotoxicity.
Effect of Varying Dosages and Adjuvants on Antibody Response in Squirrel Monkeys (Saimiri sciureus) Immunized With the Porcine Zona Pellucida Mr = 55,000 Glycoprotein (ZP3)
Authors: SACCO ANTHONY G. , YUREWICZ EDWARD C. , SUBRAMANIAN MARAPPA G.
Published: May 2013

ABSTRACT: Using the porcine zona pellucida Mr = 55,000 (ZP3) glycoprotein as immunogen, the effect of varying ZP3 dosages (50 μg, 25 μg, 5 μg) or adjuvants (200 μg ZP3 with either Freund's, Al[OH]3, or a muramyl dipeptide analog [MDP] on antibody response was evaluated in female squirrel monkeys. Although the dosage range investigated did result in dose‐dependent titration profiles, antisera from all dosage groups demonstrated in vitro contraceptive potential. MDP was as effective an adjuvant as Freund's in eliciting high titers of antibodies demonstrating contraceptive potential. Al(OH)3 was not an effective adjuvant in this system. Fertility data, utilizing those monkeys in the dosage study, suggested that an RIA titer >40% binding was necessary to prevent pregnancy. We conclude that in the squirrel monkey system, a 50 μg dosage of ZP3 should be adequate for inhibiting fertility and that MDP would serve as a satisfactory alternative adjuvant for Freund's.
Enzyme‐Linked Immunosorbent Assay (ELISA) for Detecting Antisperm Antibodies in Mice With Testicular Autoimmunity
Authors: ITOH MASAHIRO , HIRAMINE CHIHARU , KOSETO MASAHIRO , HOJO KENJI
Published: May 2013

ABSTRACT: We developed an ELISA for measuring antisperm antibodies in the mouse by using serum samples obtained from mice immunized with murine testicular antigens in complete Fre‐und's adjuvant (CFA) as well as from mice rendered vasectomized. Sperm antigens used were syngeneic epididymal spermatozoa and two types of soluble, murine testicular antigens prepared in our laboratory.

This study deals with a) the sequential changes of antisperm antibody levels following immunization; b) determination of immunoglobulin classes of these antibodies; c) a correlation between the absorbance values and the endpoint titers of antisperm antibodies; and d) comparison of end‐point titers of antisperm antibodies detected by ELISA with those by immunoperoxidase staining method in immune and nonimmune sera. It is suggested that serum dilution as high as 1/800 or more is required for detecting antibody titers of immune sera, because nonimmune mouse sera reveal a definite, although low, level of absorbance value at a serum dilution of 1/400 or less.
Antiphospholipid Antibodies and Reproduction
Authors: TRIPLETT DOUGLAS A. , HARRIS E. NIGEL
Published: May 2013

ABSTRACT: Antiphospholipid antibodies (APAs) may be identified in the laboratory by using either coagulation studies or solid‐phase immunologic assays (ELISA; RIA). These methodologies do not necessarily evaluate the same antibody; consequently, it is appropriate to screen a patient's plasma by utilizing both assays. APAs have been associated with a variety of obstetrical complications including recurrent spontaneous abortion, intrauterine fetal death, early onset preeclampsia, deep vein thrombosis, and postpartum serositis syndrome. The Kaolin Clotting Time appears to be the most sensitive coagulation test for identifying the lupus anticoagulant. However, preliminary studies would suggest the presence of anticardiolipin antibodies as detected by solid‐phase assays are more sensitive and predictive of the clinical course. Although there are no prospective trials to analyze treatment of patients with APA, preliminary data suggest the use of prednisone in combination with aspirin significantly improves the probability of delivery of a viable infant. In addition, heparin, intravenous gammaglobulin, and exchange plasmapheresis have all been tried with varying degrees of success in individual patients in small series.
Sperm Antibodies After Intraperitoneal Insemination of Sperm: A Preliminary Report
Authors: CRITSER E.S. , VILLINES P. , GENTRY M. , GENTRY W. , PETERS A. , COULAM C.B. , CRITSER J.K.
Published: May 2013

ABSTRACT: To test the hypothesis that intraperitoneal insemination of sperm induces the expression of anti‐sperm antibodies a prospective study was designed. Fifteen women undergoing intraperitoneal insemination (with or without oocyte transfer) were studied with 11 women having evaluation of anti‐serm antibodies. Sperm antibodies were detected by the immunobead test prior to intraperitoneal insemination and after each treatment cycle. Two criteria were used to assess positivity: the first based upon negative controls and the second based upon the evaluation of 20 fertile control couples. Using the first criteria, of 11 of the women undergoing IPI for the first time, 7 were initially negative and 4 were initially positive for at least 1 isotype. After treatment, 3 additional patients were positive (for a total of 7) and 4 patients remained negative. This alteration in sperm antibody frequency was not different (P = .4) as determined by the Fisher's Exact Test. Four of the 11 patients underwent a second cycle of IPI. All 4 patients were negative prior to the first treatment and 3 were negative prior to the second treatment. Subsequent to the second exposure, all 4 of these women were positive for at least 1 isotype. This shift in frequency distribution after 2 cycles was significant (P = .01). The frequency of antisperm antibodies for the same 11 patients was evaluated by using fertile control values as the basis of positivity. Two patients (18%) were positive for anti‐sperm antibodies prior to intraperitoneal insemination. There was no change in the frequency of positivity after 1 cycle of IPI. After 2 cycles of IPI 1 patient previously shown to be negative for anti‐sperm antibodies was subsequently positive. This shift in frequency was not significant (P = 0.1).
Comparison of the Direct Radiolabeled Antiglobulin Assay and the Direct Immunobead Binding Test for Detection of Sperm‐Associated Antibodies
Authors: HAAS GILBERT G. , LAMBERT HOVEY , STERN JUDY E. , MANGANIELLO PAUL
Published: May 2013

ABSTRACT: Thirty‐seven semen samples were assayed for sperm‐associated IgG and IgA using the immunobead test. Portions of these sperm samples were sent for testing with a direct radiolabeled antiglobulin assay and the testing results were compared. If the results of the immunobead test when only tail‐tip bead binding was noted are regarded as negative, there was close correlation between the two assay methodologies.
Altered Humoral Immunoregulation During Human Pregnancy
Authors: BISSET L.R. , FIDDES T.M. , GILLETT W.R. , WILSON P.D. , GRIFFIN J.F.T.
Published: May 2013

ABSTRACT: The in vitro production of immunoglobulins in response to stimulation with pokeweed mitogen (PWM) and fixed/killed Staphylococcus aureus Cowan 1 (SAC) was measured in conjunction with in vivo assays of plasma immunoglobulin levels to examine the quality and quantity of humoral immunity during human pregnancy and at parturition. Following stimulation with PWM, there is a significant enhancement of in vitro immunoglobulin‐G (IgG) production during pregnancy. Following stimulation with PWM and SAC, there was a significant reduction in in vitro immunoglobulin‐M (IgM) production immediately following parturition. There was a significant decrease in the plasma levels of IgG during pregnancy, although no change in the plasma levels of IgM were observed. The decrease in plasma immunoglobulin levels during pregnancy cannot be explained as the result of hemodilution and transplacental transfer. Altered humoral immunoregulation is the most likely means whereby an increase in immunoglobulin production during human pregnancy could occur. The possible effects of this on the outcome of pregnancy are discussed.