bluuebunny/crossref_metadata_2025 huggingface dataset:
Aspirin‐Interleukin‐3 Interrelationships in Patients With Anti‐Phospholipid Syndrome
Authors: Fishman Pnina , Falach‐Vaknin Emily , Sredni Benjamin , Meroni Piere Luigi , Tincani Angela , Dicker Dror , Shoenfeld Yehuda
Published: September 2011

PROBLEM: Previously we reported on the generation of experimental anti‐phospholipid syndrome (APS) in mice. These models were employed to evaluate the efficacy of various novel therapeutic modalities including interleukin‐3 (IL‐3) and low dose aspirin. The efficacy of the latter was found to be interrelated. Low dose aspirin is capable of inhibiting the activity of the enzyme cyclooxygenase which is responsible for the metabolism of arachidonic acid towards the production of prostaglandins. This shifts the metabolism of arachidonic acid to the other pathway and leads to an overproduction of leukotrienes. The leukotrienes act as stimulators of IL‐3 production, a positive cytokine in pregnancy which enhances placental and fetal development. In the current study we evaluated the IL‐3 levels in pregnant women with APS and expanded our knowledge on the interrelationships between aspirin, arachidonic acid metabolites and IL‐3 in the human system.

METHODS: IL‐3 levels were recorded in the serum of pregnant women with APS and compared to a control pregnant group. In addition peripheral blood mononuclear cells from healthy subjects were incubated with different concentrations of aspirin or with arachidonic acid metabolites (Leukotriene B4, C4 or PGE2), and IL‐3 production in the culture fluids was evaluated.

RESULTS: Serum level of IL‐3 in pregnant patients with primary APS, APS secondary to SLE and SLE was lower in comparison to the control group. The in vitro studies revealed that only low dose aspirin (10 mg/μl) stimulated IL‐3 production while higher doses of the drug failed to induce the cytokine generation. Leukotriene B4 and C4 were stimulatory whereas PGE2 acted as inhibitor of IL‐3 production.

CONCLUSIONS: The serum level of IL‐3 is decreased to pregnant women with primary or secondary APS. Low dose aspirin is capable of stimulating EL‐3 production in vitro most probably through an elevation of leukotriene production, which may explain its beneficial activity in preventing the manifestations of APS.
Role of the Major Histocompatibility Complex Region in Reproduction, Cancer, and Autoimmunity
Authors: Gill ThomasJ.
Published: September 2011

PROBLEM: A diverse body of evidence indicates that there is an association among reproductive failure, development, cancer, and autoimmunity. The evidence for the relationship of genes linked to the major histocompatibility complex to these diseases will be evaluated.

METHODS: The published results of both experimental studies in animals and of clinical observations in humans will be summarized and analyzed.

RESULTS: Experimental studies in the rat have demonstrated a relationship between the MHC‐linked region and growth, development, and susceptibility to chemical carcinogens. Data from other species support these observations. Clinical studies in humans have demonstrated an association between the MHC region and recurrent spontaneous abortions and a variety of other diseases; between recurrent spontaneous abortion and several autoimmune diseases; and between isolated populations and deficiencies of HLA homozygotes.

CONCLUSIONS: Genes in the MHC‐linked region play an important role in the control of reproduction, growth and development, and susceptibility to a variety of diseases.
Altercation of Generations: Genetic Conflicts of Pregnancy
Authors: Haig David
Published: September 2011

Pregnancy is traditionally viewed as a harmonious collaboration between mother and fetus. From this perspective, viviparity poses a series of problems that maternal and fetal genes work together to solve and the many complications of pregnancy are interpreted as evidence of the malfunctioning of an evolved system or of the failure of natural selection to achieve an adaptive goal. This view fails to recognize aspects of genetic conflict that lie at the heart of gestation. At least three interrelated sources of conflict can be identified: (i) conflict between genes expressed in the mother and genes expressed in the fetus/placenta (parent‐offspring conflict); (ii) conflict between maternally‐derived and paternally‐derived genes within the fetal genome (genomic imprinting); and (iii) conflict between maternal genes that recognize themselves in offspring and the rest of the maternal genome (gestational drive).
Functions of Human Decidual NK Cells
Authors: King Ashley , Jokhi P.P. , Burrows Tanya D. , Gardner Lucy , Sharkey A.M. , Lore Y.W.
Published: September 2011

The main population of lymphocytes found in the human decidua during early pregnancy are NK‐like cells with a distinctive phenotype, CD56bright CD16− CD3−. These cells are in close association with invading trophoblast that may be their in vivo target. We have examined three aspects of decidual NK function in vitro: cytotoxicity, proliferation, and cytokine production. The functional assays indicate uterine lymphocytes differ fundamentally from both PBL and even from classical circulating NK cells. Their role in the establishment of normal pregnancy remains unknown.
Effect of Platelet‐Activating Factor (PAF) on Preimplantation Mouse B6D2F1/J Embryo Formation
Authors: Roudebush William E. , Duralia David R. , Butler William J.
Published: September 2011

Platelet‐activating factor (1‐O‐alkyl‐2‐acetyl‐sn‐glycero‐3‐phosphocholine; PAF) is a potent signaling phospholipid that has been implicated in a variety of reproductive processes. Human, rabbit, and mouse preimplantation embryos produce and secrete PAR Anti‐PAF antibodies interfere with mouse preimplantation development. A controversy exists on whether exogenous PAF is beneficial to preimplantation embryo development. The study objective was to determine the effect of exogenous PAF on embryo formation. One‐cell mouse B6D2F1/J embryos were collected from PMSG/hCG primed females mated with fertile males. Embryos were exposed to PAF (0–10 μM) in MEM (0.3% BSA) for 15 min, then cultured in MEM (0.3% BSA) in a 5% CO2 in air, 95% relative humidity at 37°C atmosphere, for 120 hr to the hatched blastocyst stage. PAF (0.1 or 0.01 μM) significantly (P < 0.05) improved preimplantation embryo development and formation in vitro. PAF at higher doses had no significant effect. Supplementation of culture medium with exogenous PAF was beneficial to preimplantation embryo development in B6D2F1/J mice.
Effects of Sperm Immobilizing Antibodies on Pregnancy Outcome in Infertile Women Treated With IVF‐ET
Authors: Shibahara Hiroaki , Mitsuo Mizumi , Ikeda Yoshikazu , Shigeta Minoru , Koyama Koji
Published: September 2011

PROBLEM: Since it was found that anti‐sperm antibodies could impair in vitro development of fertilized eggs in the presence of complement in rats, the effects of sperm immobilizing antibodies on human pregnancy were examined in infertile women treated with IVF‐ET.

METHOD: The pregnancy outcomes of 143 ET cycles in 58 infertile women with sperm immobilizing antibodies and 363 ET cycles in patients with tubal infertility as control were compared. Diagnosis of chemical pregnancy was done when the urinary hCG level had risen over 50 IU/L but a gestational sac could not be demonstrated later. Antibody titers of sperm immobilizing antibodies (SI50 units) were measured by a quantitative sperm immobilization test.

RESULTS: 33 (23.1%) of 143 cycles in the patients with sperm immobilizing antibodies and 56 (15.4%) of 363 cycles in the control patients were diagnosed as pregnancy. The pregnancy rates were significantly higher in the former than in the latter (P<0.05). In the patients with sperm immobilizing antibodies, 12 (36.4%) were chemical pregnancies, 5 (15.2%) were clinical abortions, and 16 (48.5%) had deliveries. In the control group, 18 (32.1%) were chemical pregnancies, 10 (17.9%) were clinical abortions including ectopic pregnancies and 28 (50.0%) had deliveries. There was no significant differences in each category. When the SI50 titers at the time of conception were considered, chemical pregnancy rates were 22.2% (4/18) in patients with SI50 titers below 10 units, but those in patients with SI50 titers above 10 were 50.0% (5/10) and above 100 were 60.0% (3/5), respectively, (P > 0.05). In four of five patients who had both chemical and clinical pregnancies, the SI50 titers at the time of conception were higher in the chemical pregnancy cycles than in the clinical pregnancy cycles.

CONCLUSIONS: Though the pregnancy rates were significantly higher in the patients with sperm immobilizing antibodies as compared to those with tubal infertility, chemical pregnancy rates were also higher in the patients with higher sperm immobilizing antibody titers. These results suggest that sperm immobilizing antibodies may cause the damage of early development of human embryos in vivo in the small number of patients with a high titer of the antibodies.
Globulin‐Enriched Protein Supplements Shorten the Pre‐Compaction Mitotic Interval and Promote Hatching of Murine Embryos
Authors: Schneider E. Gayle , Hayslip C. Calvin
Published: September 2011

PROBLEM: To determine whether Synthetic Serum Substitute (SSS), which contains human globulins in addition to Human Serum Albumin (HSA), is superior to HSA alone as a protein supplement for embryo culture.

METHOD: Development of mouse zygotes to eight‐cell/compacting morulae and to hatching/hatched blastocysts was assessed in Human Tubal Fluid (HTF) medium containing either SSS or HSA.

RESULTS: Although there was no difference in the overall blastocyst rate at 120 h in HTF+SSS versus HTF+HSA, significantly more embryos at 54 h were at the eight‐cell/compacting morula stage in HTF+SSS. At 120 h, there were more hatching/hatched blastocysts in HTF+SSS, and hatching correlated with SSS concentration. Addition of isolated globulins to HSA significantly stimulated the number of hatching/hatched blastocysts. Hatching could be “rescued” by transfer of embryos grown in HTF+HSA to globulin‐containing media and prevented by removal of globulins as late as the compacted morula stage (54 h).

CONCLUSIONS: SSS is superior to HSA alone for embryo culture. The stimulatory effects on mitosis and hatching may be mediated directly by globulins or by other components in the globulin‐enriched fraction.
Susceptibility to Murine Experimental Autoimmune Oophoritis Is Associated With Genes Outside the Major Histocompatibility Complex (MHC)
Authors: Nair Sharmila , Caspi Rachel R. , Nelson Lawrence M.
Published: September 2011

PROBLEM: Neonatal thymectomy induces experimental autoimmune oophoritis in certain strains of mice, and this serves as a model for human autoimmune oophoritis. Because strong MHC associations have been noted in human autoimmune conditions, we investigated the role of MHC in determining susceptibility to murine experimental autoimmune oophoritis. Strain A mice are highly susceptible to post‐thymectomy autoimmunity, whereas strain B10 mice are relatively resistant. The availability of congenic strains of mice makes it possible to separate the effects of genetic background and specific H‐2 haplotype.

METHODS: We neonatally thymectomized A and B10 background female mice, and their H‐2 congenic counterparts, and then evaluated the resulting ovarian disease at age 6 weeks. RESULTS: A. By mice, which have the A background and the H‐2b haplotype, developed severe disease equivalent to strain A mice. Similarly, B10.A mice, which have the B background and the H‐2a haplotype, failed to develop disease. Thus, H‐2a haplotype did not convey disease susceptibility.

CONCLUSIONS: Our findings suggest that immune‐regulatory regions outside the H‐2 locus play an important role in determining susceptibility to murine post‐thymectomy autoimmune oophoritis. This is in accord with our previous findings in women that showed no association between MHC and premature ovarian failure. Thus, in this respect this model is similar to human autoimmune ovarian failure. This suggests that the non‐MHC genes conveying susceptibility to autoimmune oophoritis in mice might represent similar predisposing genes for premature ovarian failure in women.
Surface Annexin II on Placental Membranes of the Fetomaternal Interface
Authors: Kristoffersen Einar K. , Matre Roald
Published: September 2011

PROBLEM: The phospholipidbinding membrane protein annexin II has been demonstrated to possess FcR activity for IgG and has been localized to the outer part of the syncytiotrophoblast cell layer. The question has arisen whether annexin II is exposed on the surface of syncytiotrophoblast cells thus enabling it to take part in the transport of IgG across the maternal barrier.

METHOD: Syncytiotrophoblast microvillous plasma membranes were analyzed by flow cytometry for annexin II as well as established surface molecules. Fresh, fixed placental tissue was preincubated with antibodies to annexin II or known trophoblast surface molecules, and analyzed by confocal laser scanning microscopy.

RESULTS: Annexin II and its subunit p11 were expressed on the surface of the syncytiotrophoblast microvillous plasma membranes as were other established surface proteins (CD46, CD59, placental alkaline phosphatase), using both flow cytometry and confocal microscopy. Annexin was not detected on the surface of viable cultured trophoblast cells.

CONCLUSION: Annexin II is exposed on the surface of syncytiotrophoblast cells as a heterotetramer together with its light chain p11. It is exposed to maternal blood and may be instrumental in IgG transport across the placental barrier by binding
Cytokines of the Human Reproductive Tract
Authors: Srivastava Maya D. , Lippes Jack , Srivastava B.I. Sahai
Published: September 2011

PROBLEM: How is it possible that the female genital tract immunologically does not reject spermatooa nor the preimplantation and nidating embryo?

METHODS: Four fluids of the human reproductive tract, i.e., human oviductal fluid (hOF), follicular fluid (FF), amniotic fluid (AF), and seminal plasma (SP) were investigated by specific ELISA for 18 cytokines. The concentrations, presence or absence of these compounds were evaluated for their possible role in the immunology of the reproductive process.

RESULTS: Stem cell factor and IL‐11 were detected in all reproductive tract fluids examined whereas large amounts of IL‐1β and IL‐1RA was found in AF and hOF. Follicular fluid revealed IL‐2. HOF contained IL 2, IL‐6, IL‐8, TNF‐α, MIP‐1α, IFN‐γ, and high levels of IL‐1β, IL‐10, IL‐1RA, and sIL‐2R. Amniotic fluid contained sIL‐2R, IL‐8, IL‐1β, IL‐1RA, IL‐6, TNF‐α, and MIP‐1α. No IL‐12 or IL‐13 was detected in hOF, follicular fluid, or amniotic fluid.

Almost no free TGF‐β1 or TGF‐β2 was found in any reproductive tract fluid except seminal plasma. Seminal plasma contained large quantities of free TGF‐β1 (9,220 ± 3,635 pg/mL) in addition to large quantities of latent TGF‐β2 (2,933 ± 2,169 pg/mL) and TGF‐β, (71,000 ± 3,240 pg/mL). Furthermore, considerable concentrations of IL‐8 (1900 ± 374 pg/mL) and sIL‐2R (350 μ/mL) exist in seminal plasma.

CONCLUSIONS: HOF contains a high level of IL‐10 (588 ± 304 pg/mL), a powerful immune suppressor which probably plays a role in regulating immune responses in the fallopian tube and possibly in the endometrial cavity.

Our observations suggest that seminal plasma with its huge content of TGFβ provides immune protection for sperm. Unfortunately, such high concentrations of TGFβ may also inhibit an immune defense in any organ in which semen is deposited.
Susceptibility of Equine Chorionic Girdle Cells to Lymphokine‐Activated Killer Cell Activity
Authors: Vagnoni K.E. , Schram B.R. , Ginther O.J. , Lunn D.P.
Published: September 2011

PROBLEM: Equine chorionic girdle cells are a subpopulation of highly invasive trophoblast cells that attach and invade the uterine epithelium on Day 35 (Day 0 = day of ovulation). These invading chorionic girdle cells form endometrial cups that are associated with a marked local maternal leukocytic response that may result in the demise of the cups at Day 120 of pregnancy. Once endometrial cups become established in the uterine wall they do not express MHC antigens, and therefore may only be susceptible to non‐MHC restricted cytotoxic cells. The susceptibility of cultured chorionic girdle cells to LAK cell cytotoxicity was tested in order to evaluate the role of this type of cytotoxicity in the life‐cycle of endometrial cup tissue.

METHODS: Chorionic girdle cells from ten Day 34 conceptuses were collected, cultured, and used as a target cell in a lymphokine activated killer assay to determine if these cells were susceptible to lymphokine activated killing.

RESULTS: Cultured chorionic girdle cells demonstrated similar in vitro morphological features and patterns of antigen expression to those seen in vivo in endometrial cups. Cultured chorionic girdle cells were susceptible to lymphokine activated killing.

CONCLUSIONS: Lymphokine activated killer cells may play a role in the degeneration of endometrial cup tissue. The potential mechanisms of regulation of uterine LAK cell activity in the horse are discussed.
Anti‐Seminal Plasma Antibodies Associated With Infertility: II. Comparative Investigation of Human Antibody Binding to Normo‐, Astheno‐, and Azoospermic Seminal Plasma Antigens
Authors: Fichorova R. , Boulanov I. , Stanislavov R.
Published: September 2011

PROBLEM: Sera from infertile patients with elevated reactivity against normozoospermic seminal plasma (NSP) have been selected to investigate human antibody binding to seminal fluid antigens present in abnormal ejaculates.

METHOD: Sera from 32 idiopathically infertile patients and 44 control sera from fertile individuals were examined by ELISA against: 1) pooled seminal plasma from asthenozoospermic ejaculates (AsthSP), 2) pooled seminal plasma from patients with aspermatogenic azoospermia (AzooSP), and 3) chromatographic fractions from NSP, AsthSP and AzooSP.

RESULTS: Of 32 patients positive for anti‐NSP antibodies, only four exhibited increased reactivity to whole AsthSP and/or AzooSP, while 14 recognized antigens of different Mr and various distributions in the corresponding chromatographic fractions.

CONCLUSION: Targets of human anti‐NSP antibodies might be lacking, less concentrated, and/or modified in AsthSP and AzooSP. These findings suggest their physiological importance and the possible relevance of the observed auto‐ and iso‐immune responses to infertility.
Quantitative Determination of IgG and IgA on Sperm From Infertile Patients With and Without Antisperm Antibodies
Authors: Hjort Tage
Published: September 2011

Immunoglobulins on patients sperm were determined by an ELISA technique, modified from the RIA described by Haas and D'Cruz (Am J Reprod Immunol, 1989; 20:37–43), i.e., suspensions of washed sperm (40, 20, 10, and 5 mill/ml) were added to a dilution of peroxydaselabelled anti‐IgG or anti‐IgA, and after incubation the supernatants were transferred to cups in microtrays, coated with IgG or IgA, respectively. After incubation and washing ELISA reaction was carried out. Known amounts of IgG and IgA were processed in the same way to obtain a standard curve, allowing the Ig on the sperm to be expressed in ng/mill sperm.

Ejaculates with increased viscosity revealed high values, irrespective of MAR‐results and in some cases the same was true for samples with large numbers of non‐spermatozoal cells (NSC). However, among 48 samples from men from infertile couples with negative MAR, >20 mill sperm/ml, normal viscosity, and a ratio between sperm and NSC >5, the median for IgG was 0.2 ng/mill sperm, 90% had ≤ 0.8 ng/mill sperm, and the highest value recorded was 1.9 ng/mill sperm. For IgA the median was 0.5 ng, 90% of the 47 samples contained ≤ 1.9 ng, and the highest value was 3.1 ng/mill sperm.

The values for sperm from patients with a positive MAR, but without free antibodies in seminat plasma, were generally within the normal range. In contrast, increased values were recorded for most of the patients with free antibodies in seminal plasma, up to 4.2 ng/mill sperm for IgG and up to 12.5 ng/mill sperm for IgA.
Lymphocyte Subsets in the Endometrium of Patients With Endometriosis Throughout the Menstrual Cycle
Authors: Mettler L. , Volkov N.I. , Kulakov V.I. , Jürgensen A. , Parwaresch M.R.
Published: September 2011

PROBLEM: Comparison and characterisation of different lymphocyte subsets in the endometrium of endometriosis patients and in healthy women on every day of the menstrual cycle with special emphasis on the CD4:CD8 ratio in the endometrium.

METHOD: Immunohistochemical staining of 253 endometrial biopsies of infertile women with and without endometriosis with Anti‐Leu4 (CD3), Anti‐Leu3a (CD4), Anti‐Leu2a (CD8), Anti‐Leu7 and Anti‐Human‐B‐cell (CD22) using the immune peroxidase reaction. Identification and counting of positive lymphocytes were performed on cryostat sections.

RESULTS: Endometrial lymphocyte subsets show equal quantity and distribution in endometriosis patients and in the control group. After a peak in the early proliferative phase the absolute number of T lymphocytes decreases while a predominance of T‐suppressor/cytotoxic T lymphocytes (CD8) compared to T‐helper/inducer lymphocytes (CD4) occurs towards the end of the menstrual cycle.

CONCLUSION: Endometrium as the potential parent epithelia of endometriosis lesions seems not to be altered in its lymphatic cell content compared to healthy women. Furthermore, endometrium is clearly characterised as part of the mucosa associated lymphatic tissue (MALT). T lymphocytes show specific quantitative changes due to different phases of the menstrual cycle.
Antiphospholipid Antibody Prevalence in Patients With IVF Failure
Authors: Kaider Brian D. , Price David E. , Roussev Roumen G. , Coulam Carolyn B.
Published: September 2011

Antiphospholipid antibodies (APAs) have been associated with reproductive wastage. The purpose of this study was to establish the prevalence of APAs in women who have had at least 12 embryos transferred during several in vitro fertilization (IVF) cycles without ensuing pregnancy. Sera from 42 women with IVF failure and 42 women who successfully conceived after IVF were tested for the presence of APAs by ELISA. Successful post‐IVF pregnancy was determined by obtaining two consecutive rising beta‐hCG levels followed by an ultrasound to confirm a viable conceptus. The sera were tested for three isotypes of antibody: IgA, IgG, and IgM against seven phospholipids: cardiolipin (CL), phosphatidyle‐thanolamine (PE), phosphatidylinositol (PI), phosphatidic Acid (PA), phosphatidyl‐glycerol (PG), phosphatidylcholine (PC), and phosphatidyl‐serine (PS). From the IVF failure group, 11/42 (26.2%) were positive for APAs. From the control group, 2/42 (4.8%) were found positive only for IgA against PE. The difference between IVF failure and successful IVF groups was significant (P=0.01). These results suggest that antiphospholipid antibodies should be considered an important marker for increased risk of IVF failure. Patients who are involved with an IVF program should be tested for the presence of APAs prior to initiation of an IVF cycle.
Amnion Epithelial Cells, in Contrast to Trophoblast Cells, Express All Classical HLA Class I Molecules Together With HLA‐G
Authors: Hammer Astrid , Hutter Heinz , Blaschitz Astrid , Mahnert Wolfgang , Hartmann Michaele , Uchanska‐Ziegler Barbara , Ziegler Andreas , Dohr Gottfried
Published: September 2011

PROBLEM: The expression of the non‐classical HLA‐G gene has been shown at the protein level on trophoblast‐derived embryonic tissue, like the extravillous cytotrophoblast. However, the presence of HLA‐G on embryoblast‐derived cells is currently controversial. The amnion epithelium is an embryoblast‐derived cell layer covering the amnion cavity and is the main source for the amnion fluid.

METHOD: The expression of HLA class I molecules was investigated by immunohistochemical, biochemical, and molecular biological methods in amnion membranes and amnion fluid.

RESULTS: Immunohistochemically, HLA‐C and occasionally also ‐B molecules as well as HLA‐A and/or ‐G molecules have been identified on amnion epithelial cells. These results were extended by Western blotting with purified amnion epithelial cells where HLA‐B and/ or ‐C, HLA‐A and HLA‐G antigens have been detected. As expected HLA‐G mRNA was detected in amnion epithelial cells. Furthermore, classical HLA molecules as well as HLA‐G were found in amnion fluid.

CONCLUSION: These results show that the amnion epithelium frequently expresses classical HLA class I molecules as well as HLA‐G. The expression of HLA‐G antigens on amnion epithelial cells and their presence in the amnion fluid, which is continually ingested by the fetus, may be particularly relevant for the induction of peripheral tolerance.
Occurrence of the Interference of Sperm‐Associated Antibodies on Sperm Fertilizing Ability as Evaluated by the Sperm‐Zona Pellucida Binding Test and by the TEST‐Yolk Buffer Enhanced Sperm Penetration Assay
Authors: FRANCAVILLA F. , ROMANO R. , SANTUCCI R. , MARRONE V. , PROPERZI G. , RUVOLO G.
Published: September 2011

PROBLEM: This study was performed to evaluate the occurrence as well as the level of the interference of sperm‐associated antibodies on fertilization process.

METHOD: Motile sperm suspensions from 28 infertile patients with high degree of autoimmunization against the sperm head were tested with the zona pellucida (ZP) binding test and with the sperm penetration assay (SPA) enhanced with TEST‐yolk buffer. Both tests were also performed using donor sperm exposed and non‐exposed to the patients' circulating sperm antibodies.

RESULTS: A low ZP‐binding was exhibited by sperm from 50% of patients with normal semen profile. All normozoospermic patients with low ZP‐binding showed circulating sperm‐antibodies with inhibitory effect on ZP‐binding, while no patient with normal ZP‐binding showed circulating sperm‐antibodies with inhibitory effect. No normozoospermic patient exhibited a negative SPA result, and only in 16% of cases the penetration index was slightly less than 2 (the lowest value exhibited by fertile controls). Circulating antisperm‐antibodies did not significantly affect the hamster egg penetration.

CONCLUSION: Even in the presence of high degree of autoimmunization against the sperm‐head, sperm fusion with oolemma is not impaired after sperm preincubation with TEST‐yolk buffer, while an impairment of the ZP‐binding is demonstrable in half cases, when non‐immunologic factors are excluded. A substantial role in this interference is likely exerted by IgG antibodies transuded from the blood into the genital tract.
Perforin‐Expressing Lymphocytes in Peripheral Blood and Decidua of Human First‐Trimester Pathological Pregnancies
Authors: Gulan Gordan , Podack Eckhard R. , Rukavina Dr. Daniel , Gudelj Lea , Rubesa Gordana , Petrovic Oleg , Johnson Peter M. , Christmas Stephen E.
Published: September 2011

PROBLEM: We have shown previously that the decidua of first‐trimester human pregnancy is heavily infiltrated with perforin‐positive cells. The aim was to detect expression of perforin in both decidual lymphocytes (DL) and peripheral blood lymphocytes (PBL) in the first trimester of pathological pregnancies: Anembryonic pregnancy and missed abortion.

METHOD: Decidual tissue from a normal pregnancy group and from pathological pregnancies was obtained by vaginal curettage. Perforin (an intracellular antigen) and the cell surface antigens CD3, CD4, CD8, CD16, CD56, CD11c, and CD45RA were quantified simultaneously by flow cytometric analysis.

RESULTS: In the missed abortion group, we found: 1) a relative decrease in the frequency of both CD4+P+ cells and CD56+P+ cells as well as the mean fluorescence intensity for perforin; 2) a relative increase of CD16+P+ PBL cells; and 3) a relative increase of CD4+ cells in PBL compared with anembryonic pregnancy and normal pregnancy. There was also a significant relative decrease in the proportion of CD4+ and CD8+ cells among perforin‐positive PBL in both anembryonic pregnancy and missed abortion.

CONCLUSION: Our results show that significant decreases in the prevalence of perforin‐positive lymphoid cells, their subpopulations, and mean fluorescence intensity for perforin are associated with pregnancy failure.
Immunotherapy Before and During Pregnancy Improves Pregnancy Outcome in Women Who Suffer from Recurrent Abortion and Did Not Benefit from Immunotherapy Before Pregnancy
Authors: Maejima Masamoto , Fujii Tomoyuki , Yamashita Takahiro , Hara Naonori , Hamai Yoko , Miki Akinori , Kozuma Shiro , Okai Takashi , Shibata Yoichi , Taketani Yuji
Published: September 2011

PROBLEM: The appropriate modality of immunotherapy with the husband's mononuclear cells in women with a history of recurrent abortion who aborted despite the immunotherapy performed before pregnancy was explored.

METHOD OF STUDY: Nineteen patients who had suffered from recurrent abortion who had received the immunotherapy only before pregnancy and had aborted were treated with further immunotherapy performed either only before pregnancy or twice: before and during pregnancy. RESULTS: In 9 out of the 19 women who received further immunotherapy before pregnancy, 2 had healthy babies and 7 aborted again. In the remaining 10 patients who received further immunotherapy twice, before and during pregnancy, 8 had healthy babies and 2 aborted again.

CONCLUSION: Our results indicate that immunotherapy performed before and during pregnancy produces a better outcome compared with that performed only before pregnancy, especially in patients who showed no benefit from the immunotherapy performed only before pregnancy.
IL‐15, a Novel Cytokine Produced by Human Fetal Membranes, Is Elevated in Preterm Labor
Authors: Fortunato Stephen J. , Menon Ramkumar , Lombardi Salvatore J.
Published: September 2011

METHOD OF STUDY: Amniochorionic membranes were cultured in an organ explant system and were stimulated with lipopolysaccharides (LPSs). Samples were subjected to reverse transcriptase‐polymerase chain reaction (RT‐PCR) using specific primers for IL‐15 and IL‐2. The localization of mRNA and protein was accomplished by in situ hybridization and immunocytochemistry. IL‐15 was measured in culture media and amniotic fluid from term and preterm gestations by enzyme‐linked immunoadsorbent assay (ELISA). RESULTS: RT‐PCR indicated the expression of IL‐15 mRNA in the amniochorion. In situ hybridization and immunocytochemistry documented that mRNA and peptide for IL‐15 are found in amnion, chorion, and decidual cells. ELISA results indicated no significant increase of IL‐15 peptides in the culture media after LPS stimulation. Maximum levels of this cytokine were seen in the amniotic fluid (AF) of women with preterm labor compared to term labor. AF levels were not higher in preterm‐labor patients with proved infection compared with those without infection. RT‐PCR‐based detection also showed the presence of two isoforms of IL‐15 mRNA known to code for two different leader peptide sequences. IL‐2 mRNA expression was not observed in the fetal membranes.

CONCLUSIONS: The presence of IL‐15 mRNA and peptide in the amniochorion and decidua and its increased presence in the AF during preterm labor suggests a possible role for IL‐15 in preterm labor. Amniochorion is also shown to possess two IL‐15 isoform leader sequences, the differential expression of which may be involved in the regulation of IL‐15 secretion.
Intravenous Immunoglobulin in the Prevention of Recurrent Spontaneous Abortion: The European Experience
Authors: Christiansen Ole B.
Published: September 2011

PROBLEM: Clinical trials of the use of intravenous immunoglobulin (IvIg) in the treatment of recurrent spontaneous abortion (RSA) in Europe were reported and discussed.

METHOD OF STUDY: A search of the published literature, combined with our own published and unpublished results, was performed.

RESULTS: Two placebo‐controlled trials have been concluded in Europe. One trial found high but equal success rates in both the IvIg and the placebo group. Another trial found that IvIg treatment may increase the success rate by 24% compared with placebo but that the result was not statistically significant, which might be due to the small number of patients. Reasons for the diverse results of the two trials were discussed.

CONCLUSION: More and larger placebo‐controlled trials of IvIg treatment in RSA are needed before drawing definitive conclusions.
Type 2 Cadherins in the Human Endometrium and Placenta: Their Putative Roles in Human Implantation and Placentation
Authors: Maccalman Colin D. , Getsios Spiro , Chen George T.C.
Published: September 2011

PROBLEM: The Cadherins are a gene superfamily of calcium‐dependent cell adhesion molecules. To date, the role(s) of the Cadherins in human implantation remains poorly defined. METHOD OF STUDY: The spatiotemporal expression of the type 2 Cadherins, known as Cadherin‐11 and cadherin‐6, in the endometrium and placenta was examined using the reverse transcriptase‐polymerase chain reaction.

RESULTS: Cadherin‐6 and Cadherin‐11 are differentially expressed in the endometrial stroma during the menstrual cycle. The switch between cadherin‐6 and Cadherin‐11 expression in the endometrial stroma occurs during the late secretory phase. Maximum Cadherin‐11 mRNA levels were observed in the decidua of early pregnancy but were markedly reduced at term. In the placenta, Cadherin‐11 is expressed in the syncytial trophoblast and extravillous cytotrophoblast columns. However, cadherin‐6 seems to be the predominant Cadherin subtype present in highly invasive extravillous cytotrophoblasts.

CONCLUSION: Cadherin‐11 and cadherin‐6 may play a central role in the formation and organization of the human endometrium and placenta.
Endometrial Leukemia Inhibitory Factor (LIF) as a Possible Cause of Unexplained Infertility and Multiple Failures of Implantation
Authors: Hambartsoumian Edouard
Published: September 2011

PROBLEM: The possible role of leukemia inhibiting factor (LIF) in unexplained infertility and multiple failures of implantation (MFI) was evaluated.

METHOD OF STUDY: By a specific and sensitive enzyme‐linked immunosorbent assay (ELISA), the in vitro endometrial LIF secretion by explant cultures from women with unexplained infertility (n = 32) was examined. Endometrial samples were obtained on either days 8–11 of the proliferative phase or days 18–21 of the secretory phase. In some subjects (n = 11) an endometrial biopsy was performed twice, both in the proliferative and in the secretory phase of the cycle. The control group consisted of fertile women (n = 17).

RESULTS: In fertile women the endometrial LIF secretion was 2.2 times higher in the secretory phase samples than in the proliferative phase samples (mean ± SEM, 3259 ± 314 pg in the proliferative phase vs. 7726 ± 1192 in the secretory phase; P < 0.05). In contrast, infertile women exhibited no such elevation of cytokine production. Moreover, in infertile women with MFI the level of LIF in the secretory phase demonstrated the tendency to decrease (mean ± SEM, 4953 ± 1125 pg Vs. 2162 ± 541 pg; P > 0.05). When the amount of cytokine secretion was compared on the same day of the cycle between the two groups of women, the LIF production in fertile women on days 18–21 of the menstrual cycle was 3.5 times greater than in the infertile women with MFI and 2.2 times greater than in women without MFI (P < 0.01 and P < 0.05, respectively). On days 8–11 of the cycle, the level of LIF in these groups did not differ significantly, however, in infertile women the range of distribution of cytokine was largely varied, demonstrating the highest amplitude of variations in subjects with MFI. Analyses of the data for women to whom LIF was examined in both phases of the cycle showed that some subjects (27%) exhibited an elevated amount of LIF in the secretory phase of the cycle. This suggests that in these cases the endometrial factor(s) may be irrelevant to infertility.

CONCLUSIONS: In the majority of infertile women there is a deregulation of LIF production in the endometrium during both the proliferative and the secretory phases of the cycle. The dysfunction of cytokine production is more profound in patients with MFI. The deregulation of endometrial LIF secretion throughout the menstrual cycle may be a possible cause of unexplained infertility and repetitive failures of implantation.
Leukemia Inhibitory Factor in Human Reproduction
Authors: Senturk Levent M. , Arici Aydin
Published: September 2011

PROBLEM: Leukemia inhibitory factor (LIF) is a pleiotropic cytokine of the interleukin‐6 family and has different biological actions in various tissue systems. Although named for its ability to inhibit proliferation of a myeloid leukemic cell line by inducing differentiation, it also regulates the growth and differentiation of embryonic stem cells, primordial germ cells, peripheral neurons, osteoblasts, adipocytes, and endothelial cells. LIF is crucial for successful implantation of the embryo in mice. Currently, there is an accumulation of data about the role of LIF in human reproduction.

METHOD OF STUDY: This review of the literature and of our studies focuses on the expression, regulation, and effects of LIF in the human endometrium, fallopian tube, and ovarian follicle.

RESULTS: Human endometrium expresses LIF in a menstrual cycle‐dependent manner. Maximal expression is observed between days 19 and 25 of the menstrual cycle, coinciding with the time of implantation. Various cytokines and growth factors induce endometrial LIF expression in vitro. LIF receptor is expressed in endometrial tissue throughout the menstrual cycle and on human blastocysts in a stage‐dependent manner. Affecting the trophoblast differentiation pathway toward the adhesive phenotype, LIF plays a role in implantation. LIF is also expressed and secreted by the epithelial cells of the fallopian tube. Its increased expression in the tubal stromal cell cultures by the inflammatory cytokines suggests a link between salpingitis and ectopic implantation in the tube. The rising follicular fluid LIF level around the time of ovulation indicates that LIF may play a role in ovulatory events, early embryonic development, and implantation.

CONCLUSIONS: There is growing evidence that LIF may be one of the entities that plays a role in human reproduction.
Preliminary Studies with Recombinant Chorionic Gonadotropin β‐Subunit Produced in Escherichia coli for Use as an Antigen in a Birth Control Vaccine
Authors: Mukhopadhyay Asok , Bhatia Prakish K. , Majumdar Subeer S.
Published: September 2011

PROBLEM: Prototype human chorionic gonadotropin (hCG) vaccines based on natural sources are unsuitable for widespread applications due to their complex manufacturing procedures, cost, and carrier‐mediated immune suppression.

METHOD OF STUDY: Wistar rats were immunized with alum‐adsorbed CGβ (recombinant), CGβ‐TT, and nCGβ (native CGβ)‐TT, whereas Bonnet monkeys were immunized with only CGβ. The anti‐hCG antibody titre in the sera obtained at different time points were quantified by radioimmunoassay. The sera of Wistar rats were characterized in terms of their affinity to hCG, bioneutralization capacity (by inhibition of hCG‐induced testosterone production in Leydig cells), and cross‐reactivity with human luteinizing hormone, human follicle‐stimulating hormone, and human thyroid‐stimulating hormone (by direct binding assays).

RESULTS: Antigen‐binding capacities of sera obtained upon immunization with CGβ were 3,080 ± 943 ng/ml (n = 6) and 3,993 ± 1,292 ng/ml (n = 4), respectively, in rats and monkeys. The analysis of data revealed that immunization of rats with CGβ produced antibodies comparable to that of CGβ‐TT and nCGβ‐TT.

CONCLUSION: The study opens up the possibility of producing pure and highly immunogenic CGβ by a recombinant DNA route, as a consistent source of antigen for birth control vaccine.
Experimental Autoimmune Prostatitis: In Vivo Induction of the Autoimmune Response to Lymphocytic Soluble Factors. Alterations at the Endocrine Metabolism Level
Authors: Diserio Gustavo P. , Nowotny Edgar
Published: September 2011

PROBLEM: In rats, immunization with male accessory gland (MAG) extract promotes experimental autoimmune vesicle prostatitis. A specific mononuclear cell‐mediated immune response and prostate androgen metabolism impairment in MAG‐immunized rats were observed. The possibility that lymphocytic soluble factors (SoFs) can regulate the local steroid metabolism in these rats directly was studied. We investigated whether the SoFs released by MAG‐sensitized lymphocytes are capable of modifying the prostatic androgen metabolism and whether they induce histologic lesions “in vivo” when they are inoculated, carried by liposomes, into untreated rats.

METHOD OF STUDY: “In vitro” enzymatic [3H]‐5α‐dihydrotestosterone bioconversion and histologic studies were performed with prostates from SoF‐treated rats (LK rats). The obtained 3α/β‐hydroxysteroid‐oxidoreductase activities showed that LK rat values were significantly lower than in controls: 79.0 ± 2.5 vs 158.7 ± 10.2 pmol/min/mg protein, respectively (P < 0.01).

RESULTS: In the histologic studies, LK rat prostates showed focalized mononuclear infiltrates of various degrees, whereas control rats showed non‐atypic modification of the gland.

CONCLUSION: These results indicate that SoFs (probably total lymphokines) contribute significantly to the pathogenesis of experimental autoimmune prostatitis, involving a biochemical relationship between immune reaction and the androgenic enzymatic inhibition in the prostate.
Effects of Antibodies to Transferrin and Alpha 2‐HS Glycoprotein on In Vitro Sperm Motion: Implications in Infertility Associated with Endometriosis
Authors: Pillai Subbi , Rust Philip F. , Howard Leslie
Published: September 2011

PROBLEM: Women with endometriosis have antibodies to endometrial transferrin and alpha 2‐HS glycoprotein in their serum and peritoneal fluid. The objective of this study was to determine whether antibodies to transferrin and alpha 2‐HS glycoprotein adversely affect sperm motility and survival.

METHOD OF STUDY: Spermatozoa obtained from normal fertile donors and washed free of seminal plasma were incubated with the medium (control), 1:2 and 1:100 dilutions of antitransferrin, 1:4, 1:8 and 1:100 dilutions of anti‐alpha 2‐HS glycoprotein, and a 1:2 dilution of antialbumin antiserum (negative control). Sperm motion characteristics in 10 μl aliquots were evaluated at 30 min, 1 hr, 2 hr, 4 hr, and 24 hr using computerized sperm motion analysis. A paired t‐test was done to analyze the effects of the various antibodies on sperm motion characteristics.

RESULTS: Antibodies to albumin failed to adversely affect sperm motility in general or the several sperm motion characteristics in particular. In contrast, antibodies to transferrin at the dilution of 1:2 adversely affected the percentage of motile and rapid spermatozoa, progressive and path velocities, straightness, linearity, track speed, and anterior‐lateral head displacement (P < 0.001) at all the time intervals, whereas a 1:100 dilution of this antiserum adversely affected these parameters only at 24 hr. Elongation and beat cross‐frequency were significantly affected at 4 and 24 hr by a 1:2 dilution of antitransferrin antiserum. The effects of anti‐alpha 2‐HS glycoprotein were more pronounced than those of antitransferrin, but they were similar. Dilutions of 1:4 and 1:8 were effective at all time intervals, whereas a 1:100 dilution was effective in reducing the track speed and the percentage of rapid cells at 24 hr (P < 0.001).

CONCLUSION: Antibodies to endometrial transferrin and alpha 2‐HS glycoprotein present in the peritoneal fluid, and possibly in the oviductal fluid, of patients with endometriosis may adversely affect postcoital sperm motility and sperm survival.
Increased Numbers of Circulating Interferon‐γ‐and Interleukin‐4‐Secreting Cells During Normal Pregnancy
Authors: Matthiesen Leif , Berg Göran , Ekerfelt Christina , Ernerudh Jan , Ernerudh Jan
Published: September 2011

PROBLEM: Systemic immune responses during normal pregnancy are suggested to deviate toward secretion of T helper (Th)2‐like cytokines.

METHOD OF STUDY: Blood samples from 11 healthy Primigravidae in their first, second, and third trimesters, and 8 weeks postpartum were examined. Peripheral blood mononuclear cells were separated and the numbers of interferon (IFN)‐γ‐ and interleukin (IL)‐4‐secreting cells were identified by using the enzyme‐linked immunospot test.

RESULTS: In all three trimesters of pregnancy, and also postpartum, the numbers of IFN‐γ‐and IL‐4‐secreting cells were significantly higher compared with nonpregnant controls (Mann‐Whitney; P < 0.001). The numbers of IFN‐γ‐and IL‐4‐secreting cells gradually increased as the pregnancy progressed compared with postpartum (Kruskas‐Wallis; P < 0.01 and P < 0.05, for IFN‐γ and IL‐4, respectively).

The ratios of IFN‐γ/IL‐4 during pregnancy and postpartum were unchanged, and also when compared with nonpregnant controls.

CONCLUSION: The results indicate a systemic up‐regulation of both Th1‐ and Th2‐like immune responses during normal pregnancy.
Hepatocyte Growth Factor in Human Breast Milk
Authors: Yamada Yoshihiko , Saito Shigeru , Morikawa Hajime
Published: September 2011

PROBLEM: The purposes of this study were to investigate the presence of hepatocyte growth factor (HGF) in human milk, to identify the cells that produce HGF in human milk, and to determine the contribution of HGF to the growth of neonates.

METHOD OF STUDY: The HGF concentrations in serum and whey were determined with an enzyme‐linked immunosorbent assay kit. The presence of HGF in whey was also examined by Western blot analysis. To determine which cells in human milk produce HGF, an immunohistochemical examination was conducted. The expression of HGF mRNA in the mononuclear cells in human milk was examined by reverse transcriptase‐polymerase chain reaction (RT‐PCR). The effects of whey and of recombinant HGF (rHGF) on DNA synthesis by a rat small intestinal cell line, IEC‐6, were examined by [3H]thymidine uptake.

RESULTS: Human colostrum whey contained 2.22 ± 1.02 ng of HGF/ml. Milk whey collected 1 month later contained 1.83 ± 1.03 ng of HGF/ml. The presence of the heterodimeric form of HGF in colostrum whey was demonstrated by Western blot analysis. HGF was detected in the cytoplasm of human milk macrophages by an immunohistochemical examination, and the RT‐PCR also revealed that HGF mRNA is expressed in the mononuclear cells of human milk. DNA synthesis by IEC‐6 cells was increased by rHGF treatment and by whey treatment. The effect of whey on DNA synthesis by IEC‐6 cells was partially, but significantly, decreased by anti‐human HGF‐neutralizing antibody treatment.

CONCLUSIONS: Human milk contains a large amount of the active form of HGF, produced by macrophages, and HGF in human milk induces the growth of intestinal cells. Our data suggest that HGF in human milk is one of the important factors regulating the growth of intestinal cells in neonates after birth.