Systemic lupus erythematosus and secondary antiphospholipid syndrome in native sisters with reduced fertility.

Objective: Systemic lupus erythematosus (SLE) and secondary anti-phospholipid syndrome (APS II) can cause increased morbidity and mortality of the fetus. We followed the course of fertility of two sisters with these two basic diseases.

Methods: In the Center for Immunology of Reproduction, we confi rmed both sisters had increased levels of some selected anti-phospholipid antibodies (against phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, DL-glycerole, anexin V, phoshatidic acid, cardiolipin, beta2-glycoprotein I), anti-nuclear, and anti-DNA antibodies.

H3K4me2 accompanies chromatin immaturity in human spermatozoa: an epigenetic marker for sperm quality assessment.

Chromatin remodeling, including histone post-translational modifications, during spermatogenesis can affect sperm quality and fertility, and epigenetic marks may therefore be useful for clinical evaluations of sperm. Together with histone hyperacetylation, the dimethylation of histone H3 on lysine K4 (H3K4me2) is also required during protamination. Accordingly, we evaluated the utilization of this epigenetic mark for the identification of sperm with decrease quality and immature chromatin.

Follicular fluid levels of anti-Müllerian hormone, insulin-like growth factor 1 and leptin in women with fertility disorders.

Unlabelled: Anti-Müllerian hormone (AMH), insulin-like growth factor 1 (IGF1) and leptin are produced in the granulosa cells of follicles and play an important role in the growth and maturation of follicles. The aim of our study was to monitor AMH, IGF1 and leptin levels in a group of healthy women and compare them to a group of women with fertility disorders. The second aim was the evaluation of biomarker levels in relation to the identified cause of infertility.

Anti-Müllerian hormone in serum and seminal plasma in comparison with other male fertility parameters.

Anti-Müllerian hormone (AMH) is a factor most associated with female fertility and especially with the ovarian reserve. AMH is also used as a parameter of fertility in men as it arises from the epithelium of the seminiferous tubules that contain Sertoli cells which produce the AMH. To investigate the relationship between AMH production and sperm related parameters we compared the AMH levels in serum and seminal plasma between a group of healthy males (n=65) and male patients (n=68) of infertile couples with semen pathology.

Effect of long-term using of hormonal contraception on anti-Müllerian hormone secretion.

Anti-Müllerian hormone (AMH) is an important factor associated with female fertility and the ovarian reserve. There are several past studies available concerning the influence of hormonal contraception (HC) on serum AMH levels. Recent studies have reported that AMH levels in women using HC can be about 30% lower compared to those not using HC.

The role of antiphospholipid antibodies (aPls) in infertile women: the long-lasting experience.

Antiphospholipid antibodies (aPls) are generally characterized as heterogeneous and non-specific autoantibodies directed against various phospholipids such as cardiolipin, ph-serine, ph-inositol, ph-acid, ph-glycerol, ph-sphyngomyelin, ph-choline, annexins, and co-factor β2-glycoprotein I. aPls occur not only during autoimmune diseases but also during infectious diseases, essential hypertension, neurological complication, metabolic diseases, some drug abuse, and transplant loss. aPls are very often found in connection with reproductive failure such as repeated pregnancy loss and/or missed abortion, intrauterine fetal death, in preeclampsia, and repeated delivery of hypothrophic fetus.

Quantification of circulating fetal DNA as a tool for potential monitoring of pregnant patients with antiphospholipid antibodies.

Apoptosis of tissues of fetal origin is thought to be one of the main sources of cell-free fetal DNA (cffDNA) in maternal circulation, impaired apoptosis is also involved in the mechanisms contributing to recurrent spontaneous miscarriages (RSM) associated with antiphospholipid syndrome (APS). The APS increases the risk for preeclampsia nine times. In preeclampsia, the elevated levels of cffDNA were described by different authors.

Some immunological properties of female saliva and its effect on sperm motility.

Background: In this study, we investigated pH, levels of IgA, IgG, IgM, lysozyme and spermagglutinating antibodies in human saliva of healthy women, with or without hormonal contraception (HC) and their effect on sperm motility.

Methods And Results: Saliva was collected immediately after waking up from 59 healthy randomly selected women. We measured pH in sera and in saliva as well as immunoglobulin G, A and M levels in saliva by radial immunodiffusion.

Hyperstimulation syndrome: the levels of inhibin A and B in sera and follicular fluids.

Ovarial hyperstimulation syndrome (OHSS) represents a serious problem encountered during in vitro fertilization (IVF). We examined 10 patients with OHSS and 50 women also hormonally stimulated in the process of IVF who had no complications. In all women, we evaluated the number of obtained oocytes and the level of inhibins A and B in sera and follicular fluid collected at the time of ovarial puncture, the day embryo transfer and on the day of positivity for hCG.

Recurrent pregnancy loss, plasminogen activator inhibitor-1 (-675) 4G/5G polymorphism and antiphospholipid antibodies in Czech women.

Problem: This study compares the frequencies of plasminogen activator inhibitor-1 (-675) 4G/5G polymorphism and its relationship with eight antiphospholipid antibodies (aPLs) in serum of 157 patients with repeated pregnancy loss (RPL).

Method Of Study: PAI-1 (-675) 4G/5G polymorphism was determined using standard PCR-RFLP method. Enzyme-linked immunosorbent assay was used for the detection of aPLs against ph-serine, ph-ethanolamine, ph-inositol, ph-DL-glycerol, phosphatidic acid, annexin V, cardiolipin, and beta2-GPI.

Screening tests of reproductive immunology in systemic lupus erythematosus.

Female patients in reproductive age with systemic lupus erythematosus and fertility complications together are observed by rheumatologists, gynecologists, and reproductive immunologists. The paper notes the presence of autoantibodies to zona pellucida, to phospholipids (phosphatidyl serine, phosphatidyl ethanolamine, phosphatidyl inositol, phosphatidyl glycerol, phosphatidic acid, annexin V, beta-2 glycoprotein I, and cardiolipin) and of isoantibodies to sperm cells. Isoantibodies to sperm cells are not significantly predominant, but autoimmunity is well expressed in IgG positivity against phosphatidyl inositol, phosphatidyl ethanolamine, phosphatidyl serine, cardiolipin, and beta-2 glycoprotein I, as well as antizona pellucida antibodies in IgG isotype.

Sperm cells induce distinct cytokine response in peripheral mononuclear cells from infertile women with serum anti-sperm antibodies.

Background And Aims: Anti-sperm antibodies in can markedly reduce the likelihood of natural conception. The etiology of this anti-sperm immunity in human females is unknown. We compared the cytokine response of peripheral blood mononuclear cells (PBMCs) from infertile patients with or without anti-sperm antibodies (ASA) and fertile women.

Urgent termination of pregnancy in pre-eclampsia and panel of antiphospholipid antibodies.

Problem: The present work was undertaken to investigate the occurence of autoantibodies to eight various phospholipids in time of urgent termination of the pregnancy (sectio caesarea) in patients in reproductive age with severe preeclamptic symptoms.

Method Of Study: Autoantibodies against annexin V, ph-serine, ph-ethanolamine, ph-inositol, ph-DL-glycerol, cardiolipin, beta2-glycoprotein I (beta2-GPI), and phosphatidic acid were studied by ELISA methods.

Results: Increased levels of IgA-beta2-glycoprotein I, IgG-beta2-glycoprotein I, IgG- anti-ph-serine, and IgG-anticardiolipin were found in sera of preeclamptic women in the time of urgent sectio caesarea when compared to the control group with physiological pregnancy.

Sperm antibodies, intra-acrosomal sperm proteins, and cytokines in semen in men from infertile couples.

Problem: The aim of this study was to investigate seminal sperm-agglutinating antibodies, intra-acrosomal proteins, sperm head abnormalities, and cytokines (IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70 TNF-alpha, and IFN-gamma) in men from infertile couples.

Method Of Study: The direct mixed anti-immunoglobulin reaction test for IgG, IgA, and IgE in semen, and immunocytochemical method using monoclonal antibodies and indirect immunofluorescence for the examination of intra-acrosomal proteins in the spermatozoa were used. Cytokines in seminal plasma were determined by multiplex immunoanalytic xMAP (LUMINEX) technology.

TH1-TH2 response and the atopy risk in patients with reproduction failure.

Problem: Enhanced TH2 activity is characteristic for atopic diseases and is observed also in physiological pregnancy. The immune causes of repeated pregnancy losses and/or repeated in vitro fertilization failure may be associated with TH2 hypoactivity. The association with frequency of atopic diseases is unclear.

Antibodies against laminin-1 and sperm, intraacrosomal proteins in semen from infertile couples.

Seminal sperm-agglutinating antibodies along with IgG antibodies against laminin-1 and intraacrosomal sperm proteins were examined in seventy-one men from infertile couples. The direct mixed antiimmunoglobulin reaction test for IgG, IgA, and the commercial ELISA method for detecting IgG antibodies against laminin-1 in seminal plasma were used. Intraacrosomal proteins in the sperm heads were detected by immunofluorescence using monoclonal antibodies.

Cell-free fetal DNA in maternal plasma during physiological single male pregnancies: methodology issues and kinetics.

Objective: To analyze methodological influences and characterize the concentrations of cell-free fetal DNA (cffDNA) circulating in maternal plasma at different gestational ages in physiological pregnancies.

Methods: We investigated 238 independent samples from single male-bearing pregnancies of different gestation age. In the other 50 pregnancies, the samples were collected three times during pregnancy (at all trimesters) to evaluate the kinetics of cffDNA.

Recurrent pregnancy loss and frequency of eight antiphospholipid antibodies and genetic thrombophilic factors in Czech women.

Problem: The aim of this study was to investigate frequencies of eight antiphospholipid antibodies (aPLs) in serum, four genetic thrombophilic factors and their mutual relation in 206 patients with repeated pregnancy loss (RPL).

Method Of Study: Enzyme-linked immunosorbent assay was used for detection of aPLs against ph-serine, ph-ethanolamine, ph-inositol, DL-glycerol, phosphatidic acid, anti-annexin V, cardiolipin, and beta2-GPI. FV 1691G>A (Leiden mutation), FII 20210G>A mutation, MTHFR 677C>T and MTHFR 1298A>C variant genotypes were determined using a melting curve analysis of the PCR amplification product detected by the fluorescence resonance energy transfer.

Leukemia inhibitory factor gene mutations in the population of infertile women are not restricted to nulligravid patients.

Objective: Leukemia inhibitory factor (LIF) is one of the key cytokines in the embryo implantation regulation. We investigated the prevalence of the LIF gene mutations in the population of infertile women that consisted of nulligravid and secondary infertile patients.

Study Design: We designed a LIF gene mutation screening method that is based on the Temperature Gradient Gel Electrophoresis (TGGE).

Anti-phospholipid antibodies against phosphatidylinositol, and phosphatidylserine are more significant in reproductive failure than antibodies against cardiolipin only.

Problem: The humoral immune response to phospholipids was investigated in women with reproductive failure [1073 women after one in vitro fertilization (IVF), 853 women after two and more IVF, 627 women after three and more repeated spontaneous miscarriages or missed abortions, 412 women after diagnostic laparoscopy] and compared with that of 391 healthy fertile women.

Method Of Study: Sera from all women in the study were tested by enzyme-linked immunosorbent assay (ELISA) for the detection of IgG, IgA, and IgM isotypes of antibodies against seven phospholipids (aPLs), i.e.

Features associated with epilepsy in the antiphospholipid syndrome.

Objective: To assess the frequency of epilepsy in primary and secondary antiphospholipid syndrome (APS); to analyze the clinical and laboratory features characterizing those with epilepsy in a cohort of 538 patients with APS; and to find associated features that would suggest risk factors for epilepsy in APS.

Methods: We analyzed the clinical features of patients with APS who had epilepsy and compared them to the clinical features of non-epileptic APS patients.

Results: Of 538 APS patients, 46 (8.

Seven antiphospholipid antibodies in HIV-positive patients: correlation with clinical course and laboratory findings.

Objective: To compare the clinical course of HIV-1-infected patients, their CD4+, CD8+ T lymphocytes, and viral loads (VL) with the levels of seven antiphospholipid antibodies (aPLs) before, during, and after the highly active antiretroviral therapy (HAART).

Patients And Methods: aPLs were examined in patients (20 men, 10 women, aged 12-64 years, median 33 years) from the AIDS center of Western Bohemia before the initiation of HAART, and two (23 patients), and five (20 patients) years later. Flow-cytometry was used for CD4+ and CD8+ T lymphocytes analysis, commercial kits were used for VL-measurements, and commercial enzyme-linked immunosorbent assay (ELISA) was used to determine serum levels of anti-beta2-glycoprotein I (GPI) of immunoglobulin G (IgG) and IgA isotypes, and anticardiolipin levels (ACA) of IgG and IgM isotypes.

Endometriosis in reproductive immunology.

Problem: Endometriosis is suggested to represent an autoimmune disorder, but what is the prevalence of autoantibodies to antigens relevant to reproduction?

Method Of Study: The humoral immune response to the women with endometriosis (stage I-II: 261 women; stage III-IV: 62 women) in serum and in peritoneal fluid was investigated compared with 101 healthy women. Enzyme-linked immunosorbent assay (ELISA) was used in all the women for the detection of seven antiphospholipid antibodies [antiphospholipid antibodies (aPLs) against cardiolipin, L-phosphatidyl (ph)-serine, ph-glycerol, ph-inositol, ph-ethanolamine, phosphatidic (ph)-acid and against beta2-glycoprotein I] of class IgG, IgA, and IgM. A passive haemmagglutination method and ELISA (BioGen) was used for assessment of antizona pellucida antibodies (aZP), tray agglutination test (TAT) and indirect mixed anti-imunoglobulin reaction test (MAR-test) for the determination of sperm antibody levels.