bluuebunny/crossref_metadata_2025 huggingface dataset:
Mode of Inheritance of HLA Haplotypes Locus A,B in Siblings of Different Sexes
Authors: KOMLOS L. , LIVNI E. , KLEIN T. , NOTMANN J. , LEVINSKY H. , HALBRECHT I. , HART J. , ZAIZOV R.
Published: May 2013

METHOD: Forty‐eight parents and 172 children were typed for class I HLA antigens, locus A,B.

RESULTS: Although the number of cases is small, we observed: (1) a significantly decreased number of sons born after a first delivery of a son, as compared to a first delivery of a daughter; (2) significantly increased sharing of maternal class I HLA antigens between the firstborn son and his brothers from higher birth orders, as compared to his sisters; and (3) HLA‐A2 antigen, which is known to be involved in HLA restricted cytotoxic reactions in the recognition of minor histocompatibility antigens, was inherited in subsequent deliveries of sons as compared to daughters in a significantly higher frequency from the paternal than from maternal HLA haplotype. The results suggest that sharing of identical maternal HLA haplotypes between brothers may aid to decrease the degree of maternal sensitization to fetal antigens, and lack of HLA‐2 antigen in maternal cells from sons as compared to daughters may avoid maternal HLA‐A2 restricted cytotoxic reactions toward the male fetus.
Assay for Recombinant and Native Human Intraacrosomal Antigen SP‐10
Authors: SHEN MINCHENG , WRIGHT RICHARD M. , CARTA GIORGIO , HERR JOHN C.
Published: May 2013

PROBLEM: To develop a method to measure a recombinant sperm protein, SP‐10, during scale up and purification for a contraceptive vaccine formulation.

METHOD: A quantitative assay method for the human intraacrosomal protein SP‐10 was developed utilizing the format of indirect capture enzyme‐linked immunosorbent assay (ELISA). A SP‐10 specific monoclonal antibody mAb, MHS‐10, was used as the capture antibody. Two recognition reagents, a rabbit polyclonal anti‐SP‐10 antisera (pAb) and a biotin‐labeled mAb, MHS‐10, were used as the recognition antibodies, respectively. A SP‐10 recombinant fusion protein consisting of 125 SP‐10 amino acids linked to glutathione transferase was used as a working SP‐10 standard. The coefficient of variance for the assay system using the rabbit pAb was in the range of 0.099 to 0.157, and for the assay system using the biotinylated mAb MHS‐10 was in the range of 0.081 to 0.084.

RESULTS: Employing biotinylated MHS‐10 as the recognition antibody, it was found that the native SP‐10 molecule had more than one MHS‐10 epitope. The concentration of SP‐10 in a pool of human sperm extracts was found to be approximately 1% of the total proteins, assayed by both of the recognition antibody systems.

CONCLUSIONS: The assay system described is useful to monitor the yield of recombinant SP‐10 during scale‐up production of the SP‐10 vaccine.
Transforming Growth Factor β as an Immunosuppressive Protein in Human Seminal Plasma
Authors: NOCERA MARYANN , CHU T. MING
Published: May 2013

PROBLEM: Human seminal plasma is known to exhibit immunosuppressive activity in seminal plasma.

PURPOSE: The purpose was to characterize immunosuppressive proteins in seminal plasma.

METHOD: Gel filtration fractions of 100 to >440 kDa were identified that inhibited DNA synthesis and killing activity of interleukin‐2 stimulated lymphocytes.

RESULTS: The fractions exhibiting immunosuppression also inhibited DNA synthesis in a mink lung cell bioassay commonly used to measure the activity for transforming growth factor β (TGF‐β). The negative growth activity was diminished by aTGF‐β neutralizing monoclonal antibody. TGF‐β was further detected in the active fractions by Western immunoblot.

CONCLUSIONS: These results identified TGF‐β as an immunosuppressive protein in human seminal plasma and may provide insight into the role of immunosuppression played by seminal plasma, such as in reproduction and neoplasia.
Lack of Expression of HLA Antigens on Immature Germ Cells From Ejaculates With Antisperm Antibodies
Authors: CASTILLA JOSE A. , GIL TERESA , RODRIGUEZ FERNANDO , MOLINA JORGE , SAMANIEGO FRANCISCO , VERGARA FRANCISCO , HERRUZO ALFONSO J.
Published: May 2013

PROBLEM: The lack of expression of HLA antigen on immature germ cells from ejaculates with antisperm antibodies has been reported.

METHOD: The expression of human leukocyte antigens on immature germ cells from ejaculates with antisperm antibodies (ASA) was investigated by indirect immunofluorescence using a panel of monoclonal antibodies (MAb) and automated flow cytometry. Patients were divided into two groups: fertile (prevasectomic; N= 10), and ejaculates with ASA (10 samples with IgG and IgA ASA, and five semen samples with only IgG ASA). ASA were detected on sperm using the direct immunobead test. After centrifuging semen samples on a Ficoll‐Hypaque gradient, round cells obtained at the gradient interface were gated by a flow cytometer. The “immature germ cell window” was defined in terms of cellular volume and granularity.

RESULTS: The percentage of gated round cells from semen samples that reacted with anti‐CD45 was always less than 5%, and with anti‐CD44 less than 3%. This lack of reactivity of gated round cells with MAb specific for leukocytes and epithelial cells suggests that they were immature germ cells. Immature germ cells were unreactive with W6/32 and anti‐β‐2‐microglobulin MAb, which suggests that these cells do not express HLA class I molecules. Similarly, no reactivity of the immature germ cells with the MAb that recognize HLA class II molecules was found. No significant differences were observed in the expression of HLA molecules on immature germ cells between the different semen samples studied: fertile, and ejaculates with ASA.

CONCLUSION: The presence of ASA in ejaculate is not associated with abnormal HLA antigen expression on immature germ cells.
GnRH Agonist Induces Suppression of Lymphocyte Subpopulations in Secondary Lymphoid Tissues of Prepubertal Female Mice
Authors: RAO L.V. , CLEVELAND R.P. , ATAYA K.M.
Published: May 2013

PROBLEM: Gonadotropin‐releasing hormone (GnRH) agonists are playing an increasing role in the medical management of a variety of diseases. Recent evidence also indicates GnRH immune system interactions.

METHOD: The present study investigated the sequential changes in lymphocyte subpopulations in secondary lymphoid tissues of prepubertal female mice in vivo following Lupron depot administration. A direct two‐color immunofluorescence staining followed by flow cytometric analysis was employed.

RESULTS: Following agonist administration, white blood cell counts decreased significantly with decreases in both granulocyte and lymphocyte counts. Blood T‐cell and B‐cell subsets were also reduced although B cells decreased more markedly. In the spleen, B cells were again reduced more than T cells. There was no selective loss of either CD4 or CD8 subpopulations at any time point, in both spleen and blood. There were no differences in the percentage of lymph node subsets except that B cells decreased in the second week.

CONCLUSION: These data indicate that GnRH agonist alters specific lymphocyte subpopulations and, therefore, have the potential for affecting immune system function in vivo.
No Detection of Interferon‐γ Activity During Early Pregnancy in the Mouse
Authors: LA BONNARDIERE CLAUDE
Published: May 2013

PROBLEM: Interferons (IFN) have been shown to be secreted by the trophectoderm of implanting embryos in different species, in particular ungulates. In the pig, a clear‐cut IFN‐γ production, the role of which is unknown, was found in the trophoblast at implantation. A murine counterpart to these IFNs has not yet been identified.

METHOD: Two sets of experiments were conducted to test the presence of IFN‐γ in the mouse conceptus. First, day 4 blastocysts were collected from Swiss mice and their antiviral activity measured in a microassay using mouse L 929 cells and vesicular stomatitis virus (VSV) in the presence or absence of anti‐IFN‐γ antibodies. In a second set of experiments, uteri from Swiss mice on days 5.5 and 6.5 of pregnancy were flushed and the resulting fluids assayed in a specific and sensitive ELISA test.

RESULTS: In the antiviral assay, no consistent IFN‐like activity was found. The viral challenge also revealed a high susceptibility of mouse blastocysts to VSV infection. By ELISA, all but two samples (N = 75), whether on 5.5 or 6.5, were found negative.

CONCLUSION: We conclude that in this rodent species IFN‐γ is most probably not involved in early maternal‐fetal interactions.
Estrogen Induces Expression of Endometrial Epidermal Growth Factor Receptors Before Implantation
Authors: KLEINSTEIN J. , WESTERMANN W. , MENNENGA K. , GUSTMANN CH. , GIPS H.
Published: May 2013

PROBLEM: Pharmacological doses of 17β‐estradiol increase epidermal growth factor binding to uterine membranes.

METHODS: To determine whether physiological elevated E2 concentrations in the preimplantation phase of rabbits and preovulatory phase of women cause epidermal growth factor (EGF) receptor expression, we performed radioligand binding studies of endometrial membranes with [125I]‐EGF.

RESULTS: Unmated rabbits revealed a binding maximum (Bmax) of 514 ± 197 fmol/mg protein for [125I]‐EGF. The Bmax values increased significantly to 1424 ± 430 fmol/mg on day I and to 2244 ± 224 fmol/mg on day 2 after mating. On day 3 the Bmax values decreased to 1409 ± 238 fmol/mg and on days 4 to 8 the binding maxima were not significantly different from the Bmax values of unmated rabbits. In ovarieetomized rabbits a significant increase of [125I]‐EGF‐binding could be mimicked 12 and 18 h after the intramuscular injection of estradiol (40 μg/kg). The estradiol‐induced increase of [125I]‐EGF Bmax values.was blocked by the parallel application of the antiestrogen tamoxifen. Progesterone (2 mg/kg) did not influence [125I]‐EGF‐binding to endometrial membranes. Membrane preparations of the human endometrium from the periovulatory phase bound significantly more [125I]‐EGF compared to endometrium of the early proliferative phase. The expression of EGF receptor mRNA was localized in the epithelial cells of the human endometrium by in situ hybridization technique.

CONCLUSIONS: These results provide insight into the regulation of the autocrine and paracrine factor EGF in the endometrium during the preimplantation period.
Enhanced Expression of Resident Leukocyte Interferon γ mRNA in Endometriosis
Authors: KLEIN NANCY A. , PÉRGOLA GABRIELA M. , RAO‐TEKMAL RAJESHWAR , DEY TAMMY D. , SCHENKEN ROBERT S.
Published: May 2013

PROBLEM: Previous studies have shown that the endometrial epithelial/stromal cell proliferative activity of endometriosis is significantly less than that of normal endometrium and that the concentration of resident stromal leukocytes is significantly greater in ectopic than in eutopic endometrium. Other work has shown that interferon γ (IFNγ), secreted by resident leukocytes, inhibits endometrial cell proliferation in vitro. Accordingly, we hypothesized that the lower proliferative activity of endometriosis may be related to enhanced resident leukocyte IFN‐y production. This study was designed to assess whether resident leukocytes in endometriosis express IFNγ mRNA and to compare this expression to that of normal endometrium.

METHODS: Biopsies of ectopic endometrium (N = 16) from women in the follicular phase and normal proliferative (N = 9) and secretory (N = 8) endometria were examined for IFNγ expression. Using monoclonal antibodies specific for CD45 (leukocyte common antigen), CD3 (a T‐cell marker) and CDI lc (a macrophage marker), leukocyte types were identified immunocytochemically, followed by in situ hybridization to examine expression of IFNγ mRNA.

RESULTS: Results demonstrated that (1) the overall concentration of T cells and macrophages expressing IFN‐γ mRNA is significantly greater in endometriosis as compared to eutopic endometrium, and (2) the percent of each leukocyte type expressing IFNγ mRNA is greater in endometriosis than in normal endometrium.

CONCLUSIONS: These findings support a possible paracrine role for resident leukocytes in regulating cell proliferation in endometriosis.
Human Embryo Associated Immunosuppressor Factor and Serum Estradiol Synergism in Ovarian Follicle Development
Authors: BOSE RATNA , KREDENTSER J. V. , MCCOSHEN JOHN A.
Published: May 2013

PROBLEM: The purpose of this study was to test whether the level of serum immunosuppressor factor (EASF) correlates with ovarian follicle development and with serum hormones during the ovarian cycle.

METHOD: One hundred seventy‐three serum samples were collected starting at the time of ovulation and 1 wk after insemination from 29 women attempting pregnancy by ovarian hyperstimulation and intrauterine insemination. After ovarian stimulation, the patients were given human chorionic gonadotropin (hCG) 38 h before insemination. Number and size of follicles were measured by ultrasound. Serum samples were tested for hCG. estradiol (E2). luteinizing hormone, follicle‐stimulating hormone (FSH), and progesterone levels. The level of EASF in patients' sera was assayed using a two‐site sandwich enzyme‐linked immunosorbent assay system. The levels of different hormones and EASF were then correlated among themselves and with the number of ovarian follicles.

RESULTS: Eleven out of 29 patients had detectable levels of serum EASF‐Patients positive for EASF had a significantly higher number ovarian follicles ≥ 13 mm and E2 levels after hCG treatment as compared to patients with no detectable EASF. The levels of EASF correlated with number of follicles (P = .001). blood E2 (P < .02) and FSH levels (P < .02). Number of follicles ≥ 13 mm correlated with E2 (P = .01) and FSH levels (P < .01) only in patients positive for EASF.

CONCLUSIONS: The levels of EASF, E2, and FSH may act synergistically in the development of ovarian follicles.
Mapping of Immunogenic Domains on Porcine Zona Pellucida 3 α and β Glycoproteins by Murine Monoclonal Antibodies
Authors: GUPTA S.K. , BAGAVANT HARINI , CHADHA K. , GUPTA M. , YUREWICZ E.C. , SACCO A.G.
Published: May 2013

PROBLEM: Immunization with zona pellucida (ZP) leads to block in fertility with variable degree of ovarian dysfunctions. To design an immunocontraceptive vaccine based on synthetic peptides of zona pellucida, it is imperative to identify and define epitopes involved in sperm binding.

METHOD: Epitopic domains recognized by monoclonal antibodies (MAbs) specific to either porcine ZP3α or ZP3β glycoproteins were delineated in an enzyme‐linked immunosorbent assay (ELISA) based on the ability of a MAb in solution to inhibit the binding of biotinylated ZP3 to another MAb coated on a microtitration plate. Immunoblot studies were carried out to determine the nature of reactive determinants. Porcine oocytes preincubated with MAbs were tested for sperm binding in vitro.

RESULTS: Out of 23 MAbs generated, 10 had specificity for ZP3α and 13 for ZP3β. By using these antibodies, eight epitopic domains on both ZP3α and ZP3β were discernible. On ZP3β, epitopic domain DI partially overlaps with DII and DV with DVI, whereas on ZP3α domains DI to DV were in close proximity with a partial overlap, suggesting the dominance of this region. All 10 MAbs against ZP3α, and 10 out of 13 against ZP3β recognized deglycosylated forms of antigens. Seven antibodies having specificities for ZP3α and ZP3β respectively recognized linear epitopes. MA‐30, having specificity for ZP3β and MA‐420 for ZP3α and recognizing linear epitopes significantly inhibit the binding of boar sperm to porcine oocytes in vitro.

CONCLUSIONS: Collectively, these studies indicate the value of utilizing MAbs for identifying and characterizing functionally significant ZP determinants. MAbs recognizing sequential epitopes will help in the elucidation of the amino acid sequence of the epitopes, which will subsequently help in design of synthetic immunocontraceptive vaccines.
Platelet‐Activating Factor Production From In Vitro and In Vivo Fertilized Murine Embryos Is Similar
Authors: RIPPS B.A. , ZHU Y.P. , BURWINKEL T.H. , KIM H.N. , BUSTER J.E. , MINHAS B.S.
Published: May 2013

PROBLEM: This study measured platelet‐activating factor (PAF) production of in vitro and in vivo fertilized murine embryos to ascertain whether site of fertilization impacted subsequent embryonic PAF production.

METHOD: Ooocytes and embryo were collected from stimulated B6D2F1 mice. PAF production throughout the preimplantation phase from the two‐celled stage through expanded blastocysts was measured and compared among normally developing embryos with the only difference being site of fertilization.

RESULTS: A striking increase in PAF production was noted for both in vivo and in vitro fertilized embryos over the four days of culture. Significantly higher (P<.001) levels of PAF production were noted with development from the two‐celled stage (47.22 ± 4.13; 44.30 ± 2.43) to expanded blastocysts (254.31 ± 24.01;255.11 ± 5.35 ng/embryo/24h)forbothin vitro and in vivo fertilized embryos, respectively. There was no significant difference (P>.05) in PAF production between the two groups of embryos.

CONCLUSIONS: Embryonic PAF production in the mouse increases in a stage‐specific manner, and specifically the site of fertilization in vitro versus in vivo does not affect PAF production in normally developing embryos.
Relationship of Sperm Antibodies in Women and Men to Human In Vitro Fertilization, Cleavage, and Pregnancy Rate
Authors: CHANG TAI‐HO , JIH MING H. , WU TSUNG‐CHIEH J.
Published: May 2013

PROBLEM: The presence of anti‐sperm antibodies (ASA) in female serum has been correlated with decreased fertilization in the in vitro fertilization (IVF) program: however, the impact of each type of ASA (IgG, IgA. IgM) is not known.

METHOD: To clarify the role of each ASA subtype, the immunobead binding technique was used to identify IgG. IgA. and IgM ASA in the female sera and on the spermatozoa from 137 couples undergoing the IVF program.

RESULTS: Couples with ASA on spermatozoa had a lower fertilization rate and lower number of transferred embryos: and IgG was the major immunoglobin involved. Couples with ASA in female sera showed significant decreases in the rates of fertilization, cleavage, and number of transferred embryos only when IgM was detected, but not IgG or IgA. However, the presence of IgA ASA in female sera was only associated with a decrease in pregnancy rate, although the number of transferred embryos was not reduced.

CONCLUSION: These findings suggest that ASA can influence the results of IVF and that the specific effect is dependent upon the subtypes of ASA.
Amniotic Fluid Interleukin‐6 Determinations Are of Diagnostic and Prognostic Value in Preterm Labor
Authors: ROMERO ROBERTO , YOON BO HYUN , KENNEY JOHN S. , GOMEZ RICARDO , ALLISON ANTHONY C. , SEHGAL PRAVINKUMAR B.
Published: May 2013

PROBLEM: The purpose of this study was to determine if amniotic fluid concentrations of the interleukin‐6 (IL‐6) are of value in diagnosis of microbial invasion of the amniotic cavity and in the prediction of failure of tocolysis, preterm delivery and perinatal morbidity and mortality.

METHOD: Amniotic fluid was obtained by transabdominal amniocentesis from 146 consecutive patients admitted with the diagnosis of preterm labor and intact membranes. Fluid was cultured for aerobic and anaerobic bacteria as well as for mycoplasmas. Amniotic fluid IL‐6 levels were measured using a monoclonal antibody‐based enzyme‐linked immunosorbent assay with a sensitivity of 0.03 ng/ml. Logistic regression and Cox's proportional hazards model were used to examine the effect of several variables on dichotomous outcomes or interval to delivery.

RESULTS: Patients with a positive amniotic fluid culture had a significantly higher amniotic fluid IL‐6 concentrations than patients with a negative culture (median 91.2 ng/ml, range 0.9 to 437 ng/ml versus median 0.4 ng/ml, range <0.3 to 195 ng/ml, respectively; P<.0001). An amniotic fluid IL‐6 concentration of greater than or equal to 11.3 ng/ml had a sensitivity of 93.3% (14 of 15) and a specificity of 91.6% (120 of 131). All patients with an amniotic fluid IL‐6 concentration above 11.3 ng/ml and a negative amniotic fluid culture (N = 11) delivered preterm and all placenta available for examination (N = 7) had histologic evidence of chorioamnionitis. Amniotic fluid concentrations of IL‐6 were an independent predictor of preterm delivery, amniocentesis‐to‐delivery interval and neonatal morbidity and mortality. Moreover, IL‐6 concentrations added significant information to the prediction of these outcomes to that provided only by clinical information such as cervical dilatation, gestational age at admission or at delivery.

CONCLUSION: IL‐6 is a sensitive and rapid test for the detection of microbial invasion of the amniotic cavity and for identifying women at risk for spontaneous preterm delivery and neonates at risk for morbidity and mortality.
Recombinant Interleukin‐2 Corrects In Vitro the Immunological Defect of Endometriosis
Authors: MELIOLI GIOVANNI , SEMINO CLAUDIA , SEMINO ANTONIO , VENTURINI PIER LUIGI , RAGNI NICOLA
Published: May 2013

PROBLEM: It has been shown recently that women with endometriosis have several immunological defects. In particular, these patients have a defect in peripheral blood natural killer cell activity and have lost their capacity of recognizing and lysing autologous endometrial cells.

METHOD: We evaluated the effect of recombinant interleukin‐2 (rIL‐2) on peripheral blood lymphocytes obtained from both healthy donors and endometriosis patients. The generation of a strong cytolytic activity against autologous endometrial cells was obtained from both normal donors and endometriosis patients.

RESULTS: Interestingly, these cytolytic cells belong to the T‐cell lineage and do not recognize both autologous keratinocytes and allogeneic endometrial cells, thus suggesting a mechanism of specific recognition of autologous cells.

CONCLUSION: The capability of restoring cytolytic activity using rIL‐2 suggests a new immunotherapeutic approach for the treatment of severe endometriosis.
Effect of Ionophore Challenge on Hamster Egg Penetration and Acrosome Reaction of Antibody‐Coated Human Sperm
Authors: ROMANO R. , SANTUCCI R. , MARRONE V. , FRANCAVILLA F.
Published: May 2013

PROBLEM: Following the demonstration that antisperm antibodies do not affect the spontaneous acrosome reactions (AR) of human sperm used for the hamster egg penetration assay (HEPA), we evaluated the effect of the ionophore challenge on HEPA and AR of antibody‐coated sperm.

METHOD: Motile sperm suspensions from donors were exposed to serum and seminal plasma samples with sperm‐head‐directed antibodies, washed, capacitated, and challenged with ionomicin.

RESULTS: The proportion of acrosome‐reacted sperm, assessed by fluoresceinated Pisum sativum agglutinin and supravital stain Hoechst 33258, was significantly higher in the antibody‐exposed sperm than in the controls (47.0 ± 13.2% vs. 39.3 ± 12.1, respectively; P < 0.05). However, the hamster egg penetration rate and index were not significantly different between antibody‐exposed sperm and controls (98.0 ± 3.7% vs.85.9 ± 16.2% and 6.64 ±4.1 vs. 6.81 ± 4.8, respectively). Only one of eight test samples with antisperm antibodies, where the highest increase of AR rate compared with controls was observed (55.0% vs. 35.6%) produced a substantial increase of penetrations per oocyte (7.4 vs. 4.1).

CONCLUSION: The ionophore challenge enabled us to recognize a subtle effect of antisperm antibodies on the dynamics of acrosomal loss, the clinical relevance of which needs to be elucidated.
Antinuclear and Antiphospholipid Antibodies in Healthy Women With Recurrent Spontaneous Abortion
Authors: Eroglu Gungor E. , Scopelitis Eve
Published: May 2013

PROBLEM: To determine the presence or absence of subclinical autoimmunity in healthy women with first trimester recurrent spontaneous abortion (RSA), the sera of 72 healthy women with a history of three or more consecutive abortions were analyzed for the presence of antibodies to phospholipid and nuclear antigens.

METHOD: Lupus anticoagulant was determined, as were antibodies to cardiolipin. An indirect immunofluorescent antinuclear test was performed and specific antinuclear antibodies to double‐stranded DNA, ribonucleoprotein, Smith antigen, and Sjogren's syndrome A and B antigens were tested.

RESULTS: Nine women (13.2%) had low levels of antinuclear antibodies, none of which were specific. Four women (5.6%) had moderate to high levels of aCL antibodies.

CONCLUSION: We conclude that the prevalance of aCLs and antinuclear antibodies in healthy women with first trimester RSA is low. Subclinical autoimmunity is not a common cause of first trimester RSA.
The Influence of Intravenous Immunoglobulin Treatment on Maternal Immunity in Women With Unexplained Recurrent Miscarriage
Authors: Maruyama Tetsuo , Makino Tsunehisa , Iwasaki Ken‐ichi , Sugi Toshitaka , Saito Suguru , Umeuchi Masakatsu , Ozawa Nobuaki , Matsubayashi Hidehiko , Nozawa Shiro
Published: May 2013

PROBLEM: Recently the protective value of high‐dose intravenous immunoglobulin (IVIG) in the treatment of unexplained recurrent miscarriage has been reported to be similar to that of conventional immunotherapy with paternal leukocytes. We examined the effect of IVIG treatment on the cellular and humoral level of maternal immunity to demonstrate the possible mechanism by which IVIG might act to prevent recurrence of pregnancy loss.

METHOD: Eight patients were treated with a 20‐ to 25‐g dose of IVIG every 2 to 3 wk during their first‐trimester pregnancies. The development of anti‐idiotypic autoantibodies against maternal T‐cell receptors, maternal anti‐paternal lymphocyte antibodies detected by flow cytometric crossmatch, and changes of maternal lymphocyte subpopulations were monitored before pregnancy and then weekly during IVIG treatment.

RESULTS: Five of eight patients gave birth successfully after IVIG treatment given during the first trimester of pregnancy (success rate: 62.5%). Although we could not demonstrate a general immunological effect of IVIG on maternal immunity in vivo, a few significant changes of immunological parameters were found in some patients.

CONCLUSION: Our results suggest that the effect of IVIG on maternal immunity is not a passive increase of blocking antibody including anti‐HLA antibody or modification of maternal T‐cell subsets but, more likely, a passive increase of anti‐idiotypic antibody against anti‐HLA antibody or soluble HLA antigens. However, whether the immunomodulating effect of IVIG is related to its possible mechanism to prevent abortion remains unestablished.
Cellular and Humoral Immune Parameters in Women With Pathological Hyperprolactinemia Before and During Treatment With Bromocriptine
Authors: López‐Karpovitch Xavier , Larrea Fernando , Cárdenas Rocio , Valencia Xavier , Piedras Josefa , Díaz‐Sánchez Vicente , Alarcón‐Segovia Donato
Published: May 2013

PROBLEM: Experimental and clinical evidence has suggested an immunostimulatory effect of prolactin and that bromocriptine, an inhibitor of prolactin release, counteracts the actions of prolactin on the immune system. The aim of this study was to determine the impact of elevated serum prolactin levels on the immune system in patients with pathological hyperprolactinemia.

METHOD: For this purpose, parameters of the cellular and humoral immune system were studied in six women with prolactinomas and one with idiopathic hyperprolactinemia. Studies were performed when serum prolactin concentrations were high as well as during different phases of the menstrual cycle when prolactin levels had been normalized through treatment with bromocriptine.

RESULTS: Hyperprolactinemic subjects, when compared with six age‐matched normal women, had significantly higher percentages of total lymphocytes and CD2 + cells. Elevation of CD4 + cells was also observed although to a lesser extent. Bromocriptine‐treated patients, when compared with normal women were characterized by increased numbers of total lymphocytes and CD4+ cells, decreased percentage of CD8+ cells, and increased concentrations of serum IgM. These last two findings were also significantly different when compared to those observed in hyperprolactinemia.

CONCLUSION: In this study we have described the changes on cellular and immune parameters in patients with hyperprolactinemia before and during bromocriptine therapy, which support the links of communications between the immune and endocrine systems in humans.
Risk Factors for Antisperm Antibodies in Infertile Men
Authors: Heidenreich Axel , Bonfig Roland , Wilbert Dirk M. , Strohmaier Walther L. , Engelmann Udo H.
Published: May 2013

PROBLEM: The prevalence of anti‐sperm antibodies (ASAs) in the general population is 0 to 2%; the prevalence in infertile men is much higher at 7 to 26%. However, the role of ASAs in male infertility remains controversial to date. Although several risk factors for ASA development have been defined (such as testicular torsion, varicocele, cryptorchidism, vasectomy, and genital tract infection), there are no specific indications for ASA testing.

METHOD: In order to examine if a single parameter exists identifying patients with elevated ASA titers, serum ASA testing was performed with an enzyme‐linked immunosorbent assay (ELISA) in 226 consecutive male patients. The new assay, synchron ELISA (Synelisa) used in our study represents a new type of ELISA without fixation of the sperm surface antigens by formaldehyde or glutaraldehyde. Therefore the quantitative assay is highly sensitive and reproducible since the structure of sperm surface antigens is not altered by the fixation process.

RESULTS: The prevalence of ASAs in this population was 14%, while the prevalence of the control group was 2.5%. Of all factors analyzed only a history of vasectomy, an acute epididymitis, and an abnormal result in the bovine mucus penetration test was associated with elevated ASA titers (P<001). In addition, we could demonstrate a time related formation of ASAs in men after vasectomy.

CONCLUSIONS:
Treatment of Antibody‐Associated Sperm With Media Containing High Serum Content: A Prospective Trial of Fertility Involving Men With High Antisperm Antibodies Following Intrauterine Insemination
Authors: Byrd William , Kutteh William H. , Carr Bruce R.
Published: May 2013

PROBLEM: Antisperm antibodies (ASAs) associated with the sperm surface can significantly influence oocyte fertilization. We initiated a prospective trial comparing the effect of serum‐medium dilution on ASA binding and/or distribution following ejaculation and on subsequent pregnancy rates following intrauterine insemination (IUI).

METHOD: Infertile couples (N = 16) were entered into this prospective randomized protocol where the husband's sperm was at least 50% positive for both IgA and IgG ASAs using the immunobead assay. Couples underwent IUI with washed sperm collected during ejaculation into (a) 10 ml of serum medium made up of 50% maternal serum (antibody negative) in Hepes‐buffered medium (SM) or (b) in a sterile cup (DRY). For the following cycle, each couple received the alternate sperm treatment. All patients underwent at least two DRY and two SM collections prior to beginning IUI. Sperm from these collections were analyzed by manual semen analysis, computer‐aided semen analysis (CASA), and immunobead testing. All sperm preparations for IUI were analyzed by manual analysis and CASA before IUI. Outcome measures were changes in the distribution and amount of sperm antibody binding, sperm motion parameters, and pregnancy rates following IUI. Statistical analysis was performed using Fisher's exact test.

RESULTS: Collection of sperm into SM significantly reduces (P < 0.01) the percentage of antibody‐bound sperm (54.8% IgA, 60.0% IgG) versus 83.5% IgA and 87.7% IgG with DRY collection. The distribution or pattern of antibody binding to the sperm also was altered by SM. There was no significant difference between the motility of the sperm following collection with SM or DRY sperm as determined by manual and CASA methods. More importantly, there was no statistically significant difference in the pregnancy rates/ treatment cycle following SM (3.1% 32 cycles) or DRY (6.7%, 30 cycles).

CONCLUSION: Our data suggests that SM collection alters immunobead detectable ASA binding. Localization of ASA binding sites suggests that head‐bound antibodies are influenced by SM treatment with little effect on tail‐bound antibodies. In spite of the significant reduction of ASAs present on the sperm following SM treatment, there was no influence on pregnancy rates.
Cytokines in Semen of Normal Men and of Patients With Andrological Diseases
Authors: Comhaire F. , Bosmans E. , Ombelet W. , Punjabi U. , Schoonjans F.
Published: May 2013

PROBLEM: The potential value of assessment of cytokine concentrations for the diagnosis of certain pathological conditions of male reproduction has not been fully evaluated.

METHOD: The concentrations of interleukin 6 (IL‐6, pg/mL), its soluble receptor (IL‐6 sR, ng/mL), and of interleukin 1β (IL‐1β, pg/mL) have been measured in semen samples of 114 men and 12 corresponding blood sera.

RESULTS: The concentration of IL‐6 was unrelated to that of its receptor. Both IL‐6 and IL‐6 sR were higher in the first (mean: 69 and 31 resp.) than in the second fraction (39 and 13) of split ejaculates, and were within normal limits in vasectomised men. The Interleukin Iβ concentration presented a strong positive correlated with that of IL‐6 (r=0.74, P<0.001). The concentrations of IL‐6 and IL‐1β were unrelated to sperm concentration, motility and morphology, and they were within normal limits in immunological cases. Both IL‐6 and IL‐1β were higher (P<0.01) in cases with accessory gland inflammation.

CONCLUSIONS: Measurement of in particular IL‐6 in semen may contribute to the diagnosis of inflammatory disease of the accessory sex glands (positive predictive value = 98%, sensitivity = 72%, specificity = 94%), but it is not relevant for the diagnosis of immunological disease.
Effect of Paternal Leukocyte Immunization on Implantation After Biochemical Pregnancies and Repeated Failure of Embryo Transfer
Authors: Carp H.J.A. , Toder V. , Mashiach S. , Rabinovici J.
Published: May 2013

PROBLEM: It is unclear how paternal leukocyte immunization prevents pregnancy loss. We investigated whether immunization affects early pregnancy, and whether it increases the rate of implantation and early embryonic development at the time of implantation. METHOD: Three groups of women with possible implantation failure were immunized: (1) recurrent biochemical pregnancies (>3), (2) three or more failed in vitro fertilization (IVF) cycles, and (3) IVF after immunization for recurrent abortion (>3). Patients were immunized if antipaternal complement‐dependent antibody (APCA) was negative, and mixed lymphocyte culture hyporeactive to husbands lymphocytes. Immunizations were prepared from 100 ml paternal blood separated by Ficoll Hypaque density centrifugation. IVF was attempted after APCA was produced.

RESULTS: In the women with biochemical pregnancies, 10 to 12 subsequent pregnancies developed normally. Seventy‐nine subsequent IVF cycles in 33 patients resulted in 20 pregnancies, 16 of which developed normally. Six pregnancies followed 30 cycles in recurrent failed IVF, and 14 pregnancies from 49 cycles in 23 patients with previous habitual abortion.

CONCLUSIONS: This treatment may prevent further biochemical pregnancies if control studies confirm this preliminary observation, but was not shown to affect recurrent failed embryo transfer.
Maturation Antigens Present in Epididymis of Cynomolgus Monkeys (Macaca fascicularis)
Authors: Mahony Mary C. , Fan Qinuo , Cartwright Suzanne , Hodgen Gary D.
Published: May 2013

PROBLEM: The present study was carried out to evaluate the changes in morphology and motility of spermatozoa retrieved from different regions of the epididymis of the cynomolgus monkey. The role of the epididymis in sperm maturation is assessed by assaying protein synthesis within different regions of the epididymis and by correlating these with changes in spermatozoal membrane surface components.

METHOD: Spermatozoa retrieved from proximal caput (CP), midcorpus (CO), and distal cauda (CD) were assessed by morphological evaluation and computerized motion analysis. Membrane surface proteins of spermatozoa of different epididymal regions were extracted and separated on SDS‐PAGE. Protein synthesis of different regions of the epididymis were assayed in vitro by [35S]‐methionine incorporation.

RESULTS: Spermatozoa obtained from different regions of the epididymis differed morphologically only in the location of the cytoplasmic droplet. Specifically, from caput to corpus to cauda, sperm steadily exhibited a more distal cytoplasmic droplet. When the motion parameters of velocity and amplitude of the lateral head were examined, CP spermatozoa were not progressively motile, and poor duration of movement was most noticeable for CO spermatozoa compared with CD spermatozoa. Membrane extracts from CP, CO, and CD epididymal monkey spermatozoa differed in only several protein bands. Three major polypeptide bands (19, 30, and 60 kD) that were absent from CP sperm were present in CO and CD sperm, with the latter showing increased intensity. Several polypeptides were lost from the sperm during epididymal transit: a 25‐kD band was lost in CD sperm; and bands at 27 kD, 50–52 kD, and 90 kD were only present for CP sperm. Additionally, regional differences exist for proteins secreted by the cynomolgus monkey epididymis. Proteins (15, 25 kD) were only secreted in the CP region; a 38‐kD protein increased in intensity from the CP to CD regions, whereas a 21‐kD protein was absent from CD‐secreted medium.

CONCLUSION: These preliminary findings permitted the identification of several “maturational antigens” for cynomolgus monkey spermatozoa. Further characterization of these antigens that are modified during epididymal transit is warranted to determine their significance in the acquisition of progressive motility and fertilizing ability by epididymal spermatozoa.
Immunomodulation of Rat Endometriotic Implant Growth and Protein Production
Authors: Nothnick Warren B. , Curry Thomas E. , Vernon Michael W.
Published: May 2013

PROBLEM: The immune system has been implicated in the pathophysiology of endometriosis. To determine if modulation of the immune system influences endometriotic implant growth and protein production, the following experiment was conducted.

METHOD: Female rats with surgically induced endometriosis were treated with either the immunosuppressive agent pentoxifylline (Pent; 5 mg/kg BW; N = 16) or a vehicle (N = 16) for 7 consecutive days, then killed. Twenty‐four hours before death, 8 animals from each group were injected intraperitoneally with the immunostimulatory agent lipopolysaccharide (LPS; 200 Hg/kg BW). At death, endometriotic implants were measured and protein production assessed by two‐dimensional polyacrylamide gel electrophoresis.

RESULT: Pentoxifylline significantly (P < 0.001) reduced endometriotic implant size (2.15 ± 0.65 mm2 vs. 17.13 ± 1.98 mm2) whereas LPS was without effect (18.32 ± 2.57 mm2 vs. 17.13 ± 1.98 mm2). Pentoxifylline also suppressed production of a portion of the proteins that comprise the implant specific group of proteins, ENDO‐2, whereas LPS induced the production of two additional ENDO‐2 proteins.

CONCLUSION: Immunomodulatory agents can modulate rat endometriotic implant growth and production of implant‐specific proteins.
A Follow‐up Study of Immunoglobulin Levels and Autobodies in an Unselected Pregnant Population
Authors: Ailus Kirsi T.
Published: May 2013

PROBLEM: To obtain a systematic view of changes in the levels of immunoglobulins (Igs), other serum proteins, and autoantibodies during pregnancy and postpartum.

METHOD: A series of 220 women were followed throughout pregnancy and four to six months postpartum.

RESULTS: Immunoglobulin G (IgG) concentration clearly decreased toward term. The concentrations of IgM and IgA decreased only slightly. In most instances autoantibody levels paralleled changes in the corresponding immunoglobulin class levels. In few cases, however, there were clear deviations from this. With respect to IgG class autoantibodies, the highest autoantibody levels were found in the postpartum specimen. IgM class autoantbody levels remained almost constant throughout the follow‐up period.

CONCLUSIONS: The intrapregnancy decrease of IgG is mainly due to hemodilution, but when the effect of hemodilution is taken into account, total amounts of IgM and IgA are increased. The results suggests that compared to IgG the regulatory mechanisms of IgM and IgA are altered during pregnancy.
Cytotoxic Cells Directed Against Placental Cells Detected In Human Habitual Abortions by an In Vitro Terminal Labeling Assay
Authors: Yokoyama Motofumi , Sano Masatoshi , Sonoda Kenzo , Nozaki Masahiro , Nakamura Gen‐Ichi , Nakano Hitoo
Published: May 2013

PROBLEM: From the clinical point of view, it has been proposed that an immunological imbalance between the mother and the fetus might exist in one of the mechanisms for human habitual abortion. However, in the definition of habitual abortion, we have no distinct immunological criteria for this clinical entity at the moment.

METHOD: We employed aborted placental cells as the target cells in an in vitro terminal labeling (IVTL) assay system, in which the cytotoxic activity of maternal peripheral blood mononuclear cells (PBMCs) against the placental cell was examined.

RESULTS: Our results showed that the cytotoxic activity of maternal PBMCs was significantly higher in the habitual aborters (the mean target cell destruction: %TCD = 34.9%, N = 14) than that in the women with a normal first trimester (the mean %TCD = 8.9%, N= 14, P<.01). The results from the IVTL assay did not correlate with other assays using paternal lymphocytes as the target cells. The surface marker analyses revealed that CD16+ cells, CD14 + cells, and CD5+ cells were involved in the cytotoxic response against the placental cells in various degrees among the cases.

CONCLUSIONS: The above evidence suggests that a variety of cytotoxic cells participate in the phenomenon of human habitual abortion.
Steroid Sex Hormones and Macrophage Function: Modulation of Reactive Oxygen Intermediates and Nitrite Release
Authors: Chao Tzu‐Chieh , Alten Pierson J. Van , Walter Robert J.
Published: May 2013

PROBLEM: In general, females have a more active immune response than do males. The effects of female sex hormones on lymphocytes have been studied extensively but their effects on macrophages are poorly understood.

METHOD: In this study, peritoneal macrophages (M0) obtained from male rats were treated in vitro with estradiol (E2), progesterone (P), testosterone (TS), or hydrocortisone (HC) and their effects on superoxide, hydrogen peroxide, and nitrite release determined.

RESULTS: At concentrations between 10−10 and 10−9 M, female and male sex hormones had no significant effect on superoxide release but, at concentrations above or below that range, these hormones stimulated the release of these reactive oxygen intermediates (ROI). In contrast, M0 treated with HC generally exhibited either unaltered or reduced ROI release.

CONCLUSIONS: These findings suggest that female sex hormones regulate ROI release by Mø in a manner not entirely shared by other steroid hormones. At most concentrations used, E2, P, TS, and HC significantly inhibited nitrite release by Mø. However, with 10−10 M of E2 or 10−9M of P, nitrite release by Mø was not affected. In the presence of anti‐TNF antibody, the amounts of superoxide and hydrogen peroxide release were moderately reduced but nitrite release was dramatically inhibited. The sensitivity of Mø to variations in the concentrations of female sex hormones may contribute to gender‐related differences in the immune response.
The Effects of Sera From Women With Spontaneous Abortions on the In Vitro Development of Early Somite Stage Rat Embryos
Authors: Abir Ronit , Ornoy Asher , Hur Hertzel Ben , Jaffe Perhija , Pinus Helena
Published: May 2013

PROBLEM: Spontaneous abortions occur in 40 to 50% of pregnancies, but the causes for some abortions, especially those that are recurrent (spontaneous), are still unknown.

METHOD: Following previous studies that demonstrated embryotoxic effects of sera from women with spontaneous abortions in preimplantation mouse embryos, we cultured 10.5‐day‐old rat embryos in sera from women after spontaneous abortions to look for specific teratogenic effects.

RESULTS: About 50% of the embryos cultured in sera from women after spontaneous abortions were malformed, as compared to 19.1 and 27.1% malformations in embryos cultured in sera from women after a normal delivery and during a normal second trimester of pregnancy, respectively. We divided the sera from women who had spontaneous abortions into high‐risk, and low‐risk sera. In the high‐risk sera from one abortion, we found 74.2% malformed embryos and in the high‐risk group from two or more abortions this rate was 81.0%. This is compared to a rate of 17.1 and 10.3% in the low‐risk sera, respectively. We have also found lower DNA and protein synthesis in the embryos cultured in high‐risk sera compared to those cultured in low‐risk and control sera. Transmission electron microscopy examination of yolk sacs cultured in high risk sera showed ultrastructural damage as represented by a lower number of microvilli and a higher number of inclusions in the entodermal cells when compared to controls. Amino acid chromatography of the serum and the concentrations of folic acid and zinc were similar in control and high‐risk sera.

CONCLUSION: It seems that the majority of sera from women with unexplained spontaneous abortions are teratogenic to rat embryos in culture. In about two‐thirds of these sera the teratogenic factor(s) seem to be present in the IgG fraction.
Association Between Familial Autoimmune Diseases and Recurrent Spontaneous Abortions
Authors: Shelton Andrea J. , Harger James H. , Dorman Janice S. , Kuller Lewis H. , Laporte Ronald E. , Gill Thomas J.
Published: May 2013

PROBLEM: To examine the aggregation of autoimmune diseases in the families of women experiencing recurrent spontaneous abortions.

METHOD: The 95 participants in this case‐control study were recruited from Magee Womens Hospital, Pittsburgh, Pennsylvania from June 1988 to May 1991. The women having recurrent spontaneous abortions (N = 45) reported at least three early fetal losses, and the controls (N = 50) reported a minimum of three pregnancies with at least two live births and no more than one induced or involuntary pregnancy loss. Data from the participants and from their first‐degree and second‐degree relatives were obtained by questionnaire and verified by a repeated interview, if necessary.

RESULTS: The prevalence of arthritis, thyroid disease, and diabetes mellitus was increased among the relatives of women having recurrent spontaneous abortions compared to normally fertile couples. Several autoimmune diseases occurred concurrently in family members of patients, but not in the family members of normally fertile couples.

CONCLUSIONS: Autoimmune diseases occur more frequently in the families of women who have experienced recurrent spontaneous abortions. Both types of diseases involve genes in the class II region of the major histocompatibility complex.