bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of Interleukin‐10 During Pregnancy Results in Neonatal Growth Retardation
Authors: RIJHSINGHANI ASHA G. , THOMPSON KRISTIN , TYGRETTE LORRAINE , BHATIA SUDERSHAN K. 
Published: September 2011

PROBLEM: Interleukin 10 is considered to be important in the survival of the fetus in murine pregnancies that are known to be at risk for fetal wastage. The function of IL‐10 in a normal pregnancy is not known.METHODS: In this report, we attempted to neutralize Interleukin 10 by administering anti IL‐10 monoclonal antibodies (mAb) to pregnant mice that have a low background risk for fetal resorptions. The first group of mice was sacrificed on gestation day 18 to study the fetal effects of anti IL‐10 administration. The second group of mice was allowed to deliver to study the effects on the neonatal outcome.RESULTS: Administration of anti IL‐10 mAb did not affect the duration of gestation or the fetal outcome. Neonates exposed to anti IL‐10 mAb in utero showed signs of transient growth deficiency starting at 4 weeks of age that spontaneously corrected by 6 weeks of age.CONCLUSIONS: Administration of anti IL‐10 mAb does not alter the duration of gestation or the fetal outcome in normal murine pregnancies; however, it appears to be associated with transient neonatal growth problems.
Monoclonal Antibodies as Direct Probes for Human Sperm Acrosome Reaction
Authors: DORJEE S. , LAI C.L.B. , LEE A. , LEE C.Y.G 
Published: September 2011

PROBLEM: To develop simple and rapid assay procedures for determining human sperm acrosome reaction under various experimental conditions.METHODS: Specific monoclonal antibodies against human sperm acrosome were generated and utilized as probes for acrosome reaction assays by direct labelling of antibodies with fluorescein‐isothiocyanate (FITC).RESULTS: Among the generated monoclonal antibodies, HS‐63 was shown to react with antigens in the acrosome content of permeabilized acrosome‐intact human sperm, but not with those of the live ones. HSA‐10 was found to recognize antigens on the surface of sperm inner acrosome and to react with acrosome‐reacted human sperm in suspension. Following a swim‐up procedure, highly motile sperm were recovered and incubated in BWW medium at 37°C for 18 h. The percentages of acrosome‐reacted sperm were determined at various incubation times by using FITC‐labeled HS‐63 or HSA‐10 as the indicators in 10 min direct immunofluorescent assay. With the FITC‐labeled HS‐63 probe, the percentage of positively stained human sperm fixed in methanol decreased significantly after 18 h of incubation from >90 to 70–80%. In contrast, positively stained live human sperm by HSA‐10 increased significantly from <1% to 15–30%. A high correlation was obtained between the use of monoclonal antibodies and PSA (Pissum sativumagglutinin) for direct assessment of human sperm acrosome reaction.CONCLUSION: In view of the simplicity in assay procedures and evaluations, these FITC‐labeled monoclonal antibodies are reliable probes for direct assessment of acrosomal status of human sperm.
Acrobeads Test as a Predictor of Fertilization In Vitro
Authors: HERSHLAG AVNER , PAINE TERRY , SCHOLL GERALD M. , ROSENFELD DAVID L. , ZHU JING ZHANG , GUHRING PATRICIA , MECEROD DEBORAH , BENOFF SUSAN , MANDEL FRANCINE S. , BENOFF SUSAN 
Published: September 2011

PROBLEM: To determine whether the results of the Acrobeads test, which measures the expression of the complement regulator molecule CD46 on the inner acrosomal membrane following the acrosome reaction, accurately identifies semen specimens that will exhibit reduced or failed fertilization following conventional IVF insemination.METHOD: The Acrobeads test was performed on semen specimens from 97 consecutive patients preparing to undergo an PVF cycle utilizing a standardized insemination protocol. Motile sperm populations were examined at 6 h and 24 h post‐isolation for sperm‐bead agglutination. Results of the Acrobeads test were compared to that of TRITC‐PSA staining in matched specimens to directly measure the spontaneous loss of acrosome content. The percentages of TRITC‐PSA‐negative sperm were determined in freshly isolated motile populations and in duplicate aliquots incubated 18 to 20 h under sperm capacitating conditions. The relationship between the results of both analyses estimating spontaneous acrosome reactions and the rate of fertilization of metaphase II oocytes was examined.RESULTS: The Acrobeads score did not correlate significantly with the rate of fertilization by insemination at 6 h or at 24 h. The negative predictive value of this test was 21.4%. There was no correlation between the Acrobeads score and the percentage of sperm undergoing a spontaneous acrosome reaction as detected by TRITC‐PSA labeling. In contrast, the increment increase in the percentage of spontaneous acrosome reactions as quantified by TRITC‐PSA staining was correlated with the fertilization rate.CONCLUSIONS: Contrary to previous reports, our prospective, double‐blinded study failed to demonstrate that the Acrobeads test can accurately predict fertilization outcome in IVF. Therefore, the routine use of this test to screen patients prior to an IVF cycle in order to select appropriate treatment (i.e., ICSI) cannot be recommended.
Changes in T, B, and NK Lymphocyte Subsets During and After Normal Pregnancy
Authors: Watanabe Mikio , Iwatani Yoshinori , Kaneda Tatsunari , Hidaka Yoh , Mitsuda Nobuaki , Morimoto Yasuhiko , Amino Nobuyuki 
Published: September 2011

PROBLEM: Pregnancy affects the maternal immune system and the clinical course of maternal diseases. Here we report the changes in the detailed lymphocyte subsets of helper T cells, suppressor T cells, CD5+ B cells, T cell receptor (TCR) αβ‐positive T cells (Tαβ cells), TCRαβ‐negative T cell (Tγδ cells), and others during and after pregnancy through to one year postpartum, and discuss the significance of the changes.METHOD: The absolute numbers of helper T cells, suppressor T cells, cytotoxic T cells, TCRαβ‐negative T cells (Tγδ cells), CD5— B cells, CD5+ B cells, and NK cell subsets were examined by two‐color flow cytometry in peripheral blood from 51 healthy non‐pregnant women, 106 healthy pregnant women, and 148 healthy postpartum women.RESULTS: In early pregnancy, the numbers of suppressor T cells and NK cells with strong cytotoxicity (NK+++ cells) increased, and the number of cytotoxic T cells decreased. In late pregnancy, the helper T cell and NK+++ cell numbers decreased. Tαβ, CD5— B and CD5+ B cells decreased during pregnancy. After delivery, helper T cells and cytotoxic T cells increased from 1 to 4 months postpartum, and suppressor T cells increased at 7 months postpartum. TCRαβ‐negative T cells increased at 4 to 10 months postpartum. Both CD5— and CD5+ B cells decreased further at 1 month postpartum, but CD5+ B cells increased markedly at 7 to 10 months postpartum.CONCLUSIONS: These data indicate that 1) early increases of suppressor T cells and NK+++ cells during pregnancy may be related to the mechanism to accept or reject the fetus in early pregnancy, respectively; 2) late decreases of helper T cells and NK+++ cells may be related to the maintenance of pregnancy: 3) postpartum increases of helper T cells, cytotoxic T cells, TCRαβ‐negative T cells (Tγδ cells), and CD5+ B cells may be related to the postpartum aggravation of autoimmune diseases; and 4) the immunological effects of pregnancy remains until about 1 year after delivery.
Low Serum M‐CSF Levels Are Associated with Unexplained Recurrent Abortion
Authors: Katano K. , Matsumoto Y. , Ogasawara M. , Aoyama T. , Ozaki Y. , Kajiura S. , Aoki K. 
Published: September 2011

PROBLEM: The purpose of this study was to determine whether the serum macrophage‐colony stimulating factor (M‐CSF) level is associated with early pregnancy loss in unexplained recurrent spontaneous abortion (RSA) patients.METHOD: We therefore compared preconceptional serum M‐CSF levels between unexplained RSA patients and controls. The former comprised 44 bed‐rest therapy patients and 43 intradermal immunization (IDI) patients receiving paternal lymphocyte therapy, who had experienced two and three or more consecutive first‐trimester pregnancy losses, respectively. The controls were 46 healthy non‐pregnant women. We also prospectively studied the association between M‐CSF levels during pregnancy and adverse pregnancy outcomes. Sera from a total of 31 pregnant women, including 16 of the bed‐rest therapy group and 15 of the IDI therapy group, were collected at the 4th, 6th, and 8th gestational weeks and were measured for M‐CSF levels, using the enzyme‐linked immunoadsorbent assay (ELISA) method established by Hanamura et al.RESULTS: Serum M‐CSF levels were significantly lower in the non‐pregnant RSA patients (460.0 ± 185.6 U/ml; mean ± SD) than in the control group (726.5 ± 134.0 U/ml) and also were lower at the 8th, but not the 4th or 6th gestational week in those patients of both the bed‐rest and IDI therapy groups whose outcome was pregnancy failure.CONCLUSION: Thus a low level of serum M‐CSF was found to be associated with unexplained recurrent pregnancy loss in both the preconceptional and conceptional phases. These results raise the possibility that M‐CSF may play an important role in the maintenance of pregnancy and that it can be used as a parameter for determining individuals at risk.
Identification of Epitopes of Monoclonal Antibodies to Porcine Zona Pellucida 3β Glycoprotein, a Homologue of the Mouse/Human Sperm Receptor
Authors: Afzalpurkar Abhijit , Gupta S.K. 
Published: September 2011

PROBLEM: Immunization with zona pellucida (ZP) glycoproteins leads to a block in fertility with a variable degree of ovarian dysfunction. To avoid autoimmmune oophoritis, synthetic peptides corresponding to B cell epitope(s) and devoid of oophoritogenic T cell epitopes as immunogens have been proposed. The main objective of the present study is to define the epitopes recognized by monoclonal antibodies (mAbs) generated against porcine ZP3β, a homologue of the designated primary sperm receptor in mice and humans.METHODS: A multipin synthetic peptides approach has been used to map the epitopes recognized by mAbs. Dodecapeptides with an overlap of 6 amino acids corresponding to a precursor pZP3β‐deduced amino acid sequence (excluding the signal sequence) were synthesized on polypropylene pins and were tested for their reactivity with mAbs by enzyme‐linked immunoadsorbent assay (ELISA). The ability of synthetic peptides corresponding to the identified epitopes to inhibit the binding of mAbs to pZP3β in a competitive inhibition ELISA was investigated to confirm the above findings.RESULTS: Reactivity of the mAbs with the pin‐bound peptides in ELISA‐identified epitopes for mAb‐451 to EEKLVF (166–171) and mAb‐462/470 to FKAPRP (250–255) amino acid residues. mAb‐30 recognized QPVWQDEGQRLR (23–34) and VICRCC (316–321) amino acid residues. Competitive inhibition with synthetic peptides encompassing the motifs corresponding to 23–34 and 316–321 for binding of mAb‐30 to pZP3β revealed the epitopic domain to be 23–34 amino acids. Synthesis of overlapping octapeptides further identified WQDE as the minimum motif for binding of mAb‐30, and the replacement of one amino acid at a time with glycine revealed tryptophan as the critical residue.CONCLUSIONS: Collectively, these results describe peptide epitopes that will help in the design of an immunocontraceptive vaccine based on synthetic peptides corresponding to pZP3β or its homologues in other species.
T‐Helper 2 Type Cytokine and Soluble Interleukin‐2 Receptor Levels in Seminal Plasma of Infertile Men
Authors: Paradisi Roberto , Mancini Rita , Bellavia Emanuela , Beltrandi Elisabetta , Pession Annalisa , Venturoli Stefano , Flamigni Carlo 
Published: September 2011

PROBLEM: The role of cell‐mediated immunity (CMI) in unexplained male infertility and impaired sperm function has been explored.METHOD OF STUDY: The presence of cytokines, namely, interleukin‐4 (IL‐4), interleukin‐6 (IL‐6), and the soluble interleukin‐2 receptor (SIL‐2R), was investigated in seminal plasma of 18 fertile and 20 infertile subjects, using specific enzyme‐linked immunoadsorbent assays. RESULTS: IL‐4 was not detected. SIL‐2R was detected, but the concentration difference between the fertile and infertile group was not significant. IL‐6 was detected with significantly higher levels in the infertile group compared to the fertile group. IL‐6 levels in seminal plasma correlated positively with leukocyte count and negatively with sperm count, motility, and morphology.CONCLUSIONS: These findings show: a) a lack of IL‐4 in seminal plasma; b) similar SIL‐2R levels in fertile and infertile seminal plasma; c) increased IL‐6 secretion in seminal plasma of infertile subjects; and d) specific correlations of IL‐6 with the main semen parameters.
The Anovulation in Female Mice Resulting from Postnatal Injections of Estrogen is Correlated with Altered Levels of CD8+ Lymphocytes
Authors: Deshpande Rohini R. , Chapman John C. , Michael Sandra D. 
Published: September 2011

PROBLEM: Injections of estradiol‐17β (E2) are known to both induce anovulation and alter lymphocyte maturation in female mice. The current study examined whether the two events are related.METHOD OF STUDY: Female (C3H/HeJ × 129J)F1 (C31) mice were injected with 20 μg of E2 from 0–3 days, or from 3–6 days, postpartum. At 8, 12, 20, 32, or 40 weeks of age, the animals were killed, T lymphocytes were characterized, and ovaries were histologically examined for the presence of corpora lutea.RESULTS: Animals injected with E2 from 0–3 days postpartum had percentages of CD8+ thymocytes and CD8+ splenocytes that were always lower than in noninjected females, and the E2‐injected animals never ovulated, even by 40 weeks of age. In contrast, animals injected with E2 from 3–6 days of age had percentages of CD8+ thymocytes and CD8+ splenocytes that, although initially lower than in control females, attained control values by 32 weeks of age. In addition, at 32 weeks of age a number of the 3–6‐day E2‐injected females ovulated, whereas at earlier ages none had. Further, injections of E2 had little effect on the percentages of CD4+ thymocytes and splenocytes in these animals.CONCLUSIONS: The results suggest that E2‐induced anovulation in C31 female mice is correlated with decreased levels of CD8+ lymphocytes, and an increased CD4+/CD8+ lymphocyte ratio.
The Regulatory Role of Transforming Growth Factor‐Beta in Activation of Milk Mononuclear Cells
Authors: Ayoub I.A. , Yang T.J. 
Published: September 2011

PROBLEM: Immunoregulatory role of milk mononuclear cells and cytokines during the involution of the mammary gland.METHOD OF STUDY: Bioassays were used to determine the levels of transforming growth factor beta (TGF‐β) and interleukin‐2 (IL‐2) and their effects on milk mononuclear cells (MMCs). RESULTS: MMCs collected from involuting glands were less responsive to Con A stimulation than peripheral blood mononuclear cells (PBMCs), and purified huTGF‐β1 inhibited the activation of both MMCs and PBMCs by Con A. Furthermore, secretions collected over a period of approximately 4 weeks into the involution period contained high levels of active TGF‐β and extremely high levels of latent TGF‐β. MMCs stimulated with Con A produced higher levels of IL‐2 than did the PBMCs of the same animals, and bovine milk TGF‐β and huTGF‐β1 significantly inhibited the IL‐2 production by MMCs. Mammary secretions of involuting glands did not contain any detectable levels of IL‐2, and they inhibited the growth of the IL‐2‐dependent bovine lymphoblastoid T cell line (BLTC), even in the presence of 10 U/ml of rBo IL‐2.CONCLUSIONS: The findings suggest that TGF‐β is a major immunoregulatory factor in the involuting mammary gland.
Membrane Phenotype and Expression of Perforin and Serine Esterases by CD3− Peripheral Blood and Decidual Granular Lymphocyte‐Derived Clones
Authors: Gudelj L. , Christmas S.E. , Laskarin G. , Johnson P.M. , Podack E.R. , Rukavina D. 
Published: September 2011

PROBLEM: Human first‐trimester pregnancy decidua were found to contain large numbers of perforin (P)‐containing cells, which varied in their membrane antigen phenotype. In this study results obtained by analyzing CD3 clones derived from human early pregnancy decidua and peripheral blood are reported.METHOD OF STUDY: Decidual tissue was obtained from vaginal termination of first trimester normal human pregnancies. CD3− clones were generated by limiting dilution cloning after the depletion of CD3+ lymphocytes. The cell membrane phenotype was determined by (low cytometry. Perforin was detected by fluorescence‐activated cell sorter (FACS) analysis of permeabilised cells. Serine esterases (SE) were identified by histochemical staining for BLT‐esterase.RESULTS: Cloned decidual cell populations retained the overall antigenic phenotype of freshly isolated decidual natural killer (NK)‐like cells. AU CD3 clones, either derived from decidua or from peripheral blood contained perforin. Serine esterases were present in every decidual clone analyzed.CONCLUSIONS: Limiting dilution cloning allows the clear‐cut analysis of homogenous subsets of decidua‐derived NK‐like clones. The presence of large amounts of perforin in all of the CD3− clones underlines the extensive transcription of the perforin gene by NK‐like lymphocytes.
Down‐Regulated Expression of Perforin‐Positive/CD16+ Cells in the Peripheral Blood Lymphocytes in the First Trimester of Pregnancy and Up‐Regulation at the End of Pregnancy
Authors: Rukavina Daniel , Podack Eckhard R. , Rubesa Gordana , Spanjol‐Pandelo Suzana , Randic Ljiljana 
Published: September 2011

PROBLEM: Immunophenotypic profiles of perforin (P)‐positive peripheral blood lymphocytes in the first trimester and at the term of human pregnancy were analyzed.METHOD OF STUDY: Perforin expression in peripheral blood lymphocyte subsets was measured by simultaneous detection of P (intracellular antigen) and cell‐surface antigens (CD3, CD4, CD8, CD16, and CD56) by flow cytometry in nonpregnant (NP) and pregnant women in the first trimester (FTP) and at the time of parturition (TP).RESULTS: The percentage of total P+ cells in peripheral blood compared to nonpregnant women was slightly lower in the FTP but significantly higher at TP. Perforin‐positive cells were significantly elevated in T lymphocyte subsets (CD3+P+, CD4+P\ CD8+P+) in both the FTP and TP groups, as was the percentage of CD56+P+ cells. Profound changes in the CD16+ subpopulation were found in the FTP group compared to both the NP and TP groups (a drastic decrease of CD16+P+ cells; CD16+ cells among P+ cells; P+ cells among CD16+ cells). A considerable part of CD3+ cells in both the FTP and TP groups are CD3+CD56+P+. The average fluorescence intensity (AFI) for P (a measure of P content per cell) was significantly decreased in FTP and increased in TP groups.CONCLUSIONS: The CD16 molecule is FcγRIIIA which is the only Fc receptor responsible for antibody dependent cell‐cytotoxicity (ADCC) of NK and T‐cells. In the first‐trimester human pregnancy this mechanism is severely down‐regulated compared to both the NP and TP groups.
Prevalence and Biological Effects of Anti‐Trophoblast and Anti‐Endothelial Cell Antibodies in Patients with Recurrent Spontaneous Abortions
Authors: Tedesco F. , Nardon E. , Pausa M. , Bulla R. , Narchi G. , Guaschino S. , Livi C. , Meroni PL. 
Published: September 2011

PROBLEM: Trophoblasts and endothelial cells represent a potential target for antibodies in women with recurrent spontaneous abortions. These antibodies have been shown to be associated with anti‐phospholipid antibodies. Are they also present in women with unexplained pregnancy losses in the absence of anti‐phospholipid antibodies?METHOD OF STUDY: The anti‐trophoblast antibodies were tested by an immunofluorescence assay on cells purified from pooled first‐trimester placentae, whereas the anti‐endothelial cell antibodies were measured by enzyme‐linked immunoadsorbent assay (ELISA) on cells isolated from the umbilical vein and were cultured to confluence. The cytotoxicity of trophoblasts was evaluated in a homologous system. The expression of adhesion molecules on endothelial cells was quantitated by ELISA using specific monoclonal antibodies, and the expression of tissue factor was quantitated by a chromogenic assay measuring the formation of factor Xa.RESULTS AND CONCLUSIONS: Complement‐fixing antibodies to trophoblast represent a better marker to discriminate patients with recurrent spontaneous abortions from controls and are cytotoxic for the target cells. Anti‐endothelial antibodies are also present in these patients and exhibit pro‐inflammatory and pro‐coagulant activities.
Antisperm Antibody Detection: 1. Methods and Standard Protocol
Authors: Dondero Franco , Gandini Loredana , Lombardo Francesco , Salacone Pietro , Caponecchia Luisa , Lenzi Andrea 
Published: September 2011

PROBLEM: Today, antisperm antibody (ASA) detection is one of the most important steps in the evaluation of male infertility. This practice is generally accepted even though there is still some disagreement about the meaning of antisperm immunity and a good deal of controversy about the test regarded as the most suitable for the detection of antibodies directed against sperm antigens. International workshops have tried to standardize universally accepted protocols. A panel of three or four methods is generally advised to provide a correct and complete screening of patients with antisperm immunity.METHOD OF STUDY: This paper reports on the results of a serum exchange workshop for the standardization of the ASA detection in the sera carried out under the auspices of the Italian Society of Endocrinology, which was used as reference laboratory. A careful description of the most widely used methods is reported and proposed as a standard protocol also on the basis of the results of the correlation studies carried out by our group is also reported.CONCLUSIONS: If the proposed methods and procedures will be accepted by the members of the Alps‐Adria Society for Immunology of Reproduction, a Study Group for Sperm Antibody Testing, could start its activity having the goal of reaching a consensus on methodology and carrying out a European serum exchange workshop.
Characterization of Antibodies Generated Against a Conserved Portion of Oviductal Glycoprotein (OGP) and Endogenous Hamster OGP and Their Ability to Decrease Sperm Binding to the Zona Pellucida In Vitro
Authors: Schmidt A. , Mavrogianis P.A. , O'Day‐Bowman M.B. , Jaffe M.C. , Verhage H.G. 
Published: September 2011

PROBLEM: The effect of antibodies generated against hamster oviductal glycoprotein (OGP) on sperm binding to the zona pellucida (ZP) was evaluated.METHOD OF STUDY: Antibodies against a 17‐amino‐acid sequence of the OGP core protein (amino acids 52–68) and the denatured hamster OGP protein were generated, characterized, and tested in an in vitro sperm binding assay.RESULTS: Sperm binding was significantly decreased (P < 0.05) when oviductal oocytes were incubated for 2 hr with 4 or 8 mg/ml of immune IgG of both antibodies when compared with normal rabbit IgG. A fluorescence assay showed binding of both antibodies to the endogenous OGP associated with the ZP of ovulated hamster oocytes. CONCLUSIONS: These results suggest that OGP may be a potential immunocontraceptive target because both antibodies significantly decreased sperm binding to the ZP of oviductal oocytes. Immunocontraception may be accomplished by attempting to generate active immunity to a recombinant OGP, to the region selected in this study (amino acids 52–68) or to some other region of the core protein.