Anti-zona Pellucida Research Articles

Acupuncture Combined with Bushen-Jianpi Decoction Ameliorates the Ovarian Function of Diminished Ovarian Reserve Rats by Regulating Phosphoinositide 3-Kinase/AKT Signaling.

This study aimed to explore the therapeutic efficiency as well as mechanism of acupuncture combined with Bushen-Jianpi decoction (BJD) to treat rats with diminished ovarian reserve (DOR). A DOR rat model was constructed using zona pellucida 3 peptide, and acupuncture, BJD, and their combination were administered as therapeutic interventions. We measured changes in the ovarian indexes, the number of follicles at all levels, the serum levels of sex hormones and immune factors, the expression levels of phosphoinositide 3-kinase (PI3K), AKT, p-AKT, and caspase-3, and the changes in the proportions of splenic T cell subtypes, including T-helper 17 (Th17), Tc17, regulatory T (Treg), CD4+, and CD8+ cells. Acupuncture combined with BJD induced a decrease in the levels of follicle-stimulating and luteinizing hormones, and the effect was greater than that elicited by BJD or acupuncture alone (P < 0.05). Additionally, this combination treatment effectively abrogated the increase in the levels of interleukin-2 (IL-2), IL-17, anti-zona pellucida antibody, and cleaved caspase-3 (P < 0.05), while promoting the regulation of IL-6 and p-AKT (P < 0.01). Furthermore, treatment with acupuncture combined with BJD restored the proportions of CD4+ cells and the CD4+ / CD8+ T cell ratio (P < 0.01), decreased the proportion of CD8+ T and Th17 cells (P < 0.01), and increased the proportions of Tc17 and Treg cells (P < 0.01). Combining acupuncture with BJD can enhance ovarian function in DOR rats. The regulation of sex hormone levels and immune function in rats may be attributed to the adjustment of the mRNA and proteins levels of PI3K, AKT, and caspase-3 in the PI3K/AKT signaling pathway, which leads to an improvement in the immune function of DOR rats.

The mechanisms of MicroRNA 21 in premature ovarian insufficiency mice with mesenchymal stem cells transplantation

Umbilical cord-derived mesenchymal stem cell (UCMSC) transplantation has been deeply explored for premature ovarian insufficiency (POI) disease. However, the associated mechanism remains to be researched. To explore whether and how the microRNA 21 (miR-21) functions in POI mice with UCMSCs transplantation, the autoimmune-induced POI mice model was built up, transplanted with or without UCMSCs transfect with the LV-hsa-miR-21-5p/LV-hsa-miR-21-5p-inhibition, with the transfection efficiency analyzed by QRT-PCR. Mice hormone secretion and the anti-Zona pellucida antibody (AZPAb) levels were analyzed, the ovarian morphological changes and folliculogenesis were observed, and the ovarian apoptosis cells were detected to evaluate ovarian function. The expression and localization of the PTEN/Akt/FOXO3a signal pathway-related cytokines were analyzed in mice ovaries.Additionally, the spleen levels of CD8 + CD28-T cells were tested and qualified with its significant secretory factor, interleukin 10 (IL-10). We found that with the LV-hsa-miR-21-5p-inhibition-UCMSCs transplantation, the mice ovarian function can be hardly recovered than mice with LV-NC-UCMSCs transplantation, and the PTEN/Akt/FOXO3a signal pathway was activated. The expression levels of the CD8 + CD28-T cells were decreased, with the decreased levels of the IL-10 expression. In contrast, in mice with the LV-hsa-miR-21-5p-UCMSCs transplantation, the injured ovarian function can be reversed, and the PTEN/AKT/FOXO3a signal pathway was detected activated, with the increased levels of the CD8 + CD28-T cells, and the increased serum levels of IL-10. In conclusion, miR-21 improves the ovarian function recovery of POI mice with UCMSCs transplantation, and the mechanisms may be through suppressing the PTEN/AKT/FOXO3a signal pathway and up-regulating the circulating of the CD8 + CD28-T cells.

Symposium 1-4 - Immunological infertility caused by gamete immunity

1. Infertility caused by anti-sperm antibodies (ASA) 1) Female infertility caused by sperm-immobilizing antibodies in the sera In some women, ASA are produced as allogenic immunity, which causes infertility. Various methods for the antibody detection have been developed, but sperm-immobilization antibodies (SI-Ab) detected by the sperm immobilization test (SIT) have the greatest influence on the development of infertility. Sperm-immobilizing antibodies are present in the female genital tract, and also in the follicle fluid.SI-Ab cause a post-coital test (PCT) negative or impaired sperm transport from the uterine cavity to the Fallopian tubes, and fertilization disorders. There is a method to quantitatively measure SI-Ab titer, SI50 unit, and treatment methods are decided based on this SI 50 value. In cases with a SI50 value of 10 or more, it is difficult to conceive with AIH, and IVF is effective after preventing antibody contamination into the culture medium. As a mechanism for producing sperm-immobilizing antibodies, it has been speculated that women with disease-specific genes may be exposed to enough amount of sperm through sexual intercourse and develop symptoms triggered by such as chlamydia infection. CD52 is considered to be a promising antigen against sperm-immobilization antibodies, and is expected to be applied for the development of contraceptive vaccines. 2) Male infertility caused by ASA that bind to ejaculated sperm In some men, ASA are produced as autoantibodies, causing infertility. In men, it is not useful for measuring serum antibodies, therefore antibodies that bind to ejaculated sperm (sperm-binding antibodies) should be detected with ImmunoSpheres○R. However, since there are antibodies that are not related to the development of infertility, the treatment strategy can be determined by confirming whether the antibody has a biological effect. Inhibitory effect on cervical sperm penetration is confirmed by PCT and fertilization disorder is examined by the hemizona assay (HZA). If HZA cannot be performed, ICSI is recommended because it is likely to be accompanied by fertilization disorder in cases where the ratio of ASA-bound sperm to total motile sperm is 80% or more. 2. Female infertility caused by anti-zona pellucida (ZP) antibodies The ZP plays an important role in terms of egg formation, fertilization, protection of the embryo. These include the recognition of sperm species of animals, the induction of acrosome reaction, and the prevention of polysperm fertilization. On the other hand, since there is a strong antibody production ability as an antigen, if an anti-ZP antibodies are produced, the function of the ZP is impaired by the antibody, and there is a possibility of developing infertility. It was reported that anti-ZP antibodies were detected at a high rate in the sera of infertile women of unknown cause by the anti-ZP antibody detection method using porcine ZP, followed by controversial reports by another investigators. Therefore, we developed a detection method using human ZP. Anti-ZP antibodies disturb the follicular development, inhibit sperm-ZP binding and hatching. For the purpose of appropriate treatments, it was shown that ICSI is effective for fertilization disorder and assisted hatching (AHA) is effective for hatching disorder, respectively.

The effect of icariin on autoimmune premature ovarian insufficiency via modulation of Nrf2/HO-1/Sirt1 pathway in mice

Primary ovarian insufficiency (POI) is a common gynecological disease. Autoimmunity is a common cause of POI. Icariin (ICA) plays a therapeutic role in many autoimmune diseases. This study aims to investigate the effect of ICA on autoimmune POI mice and its effect on immune regulation. Sixty-three female BALB/c mice were randomized into three groups (control, POI, POI + ICA). POI and POI +ICA group were hypodermically injected with zona pellucida three peptides (pZP3) to induce autoimmune POI. Then the POI +ICA group was gavaged with ICA. A vaginal smear was to observe estrous cycles, hematoxylin-eosin staining was to count follicles. Enzyme-linked immunosorbent analysis determined serum FSH, LH, AMH, and anti-zona pellucida antibody (AZPAb) levels. In addition, flow cytometry detected the expression of Th1 cells and Treg cells, and Western blot was used to detect the expression of Nuclear factor E2 related factor 2(Nrf2), heme oxygenase-1 (HO-1), and Sirtuin-1 (Sirt1) proteins. pZP3 treatment decreased serum AMH levels and increased FSH, LH, and AZPAb levels. Additionally, decreases in the number of healthy follicles at all stages and an increase in the number of atretic follicles. Abnormal ovarian structure and an arrested estrous cycle were also noted. However, ICA rescued POI through up-regulating Nrf2, HO-1, and Sirt1 expressions and up-regulating Treg expressions. ICA treatment improved the structure of the injured ovarian and its function in autoimmune POI mice. The mechanism is achieved by increasing the expression of Nrf2/HO-1/Sirt1 pathway in the ovary and increasing Treg cells' expression.

Adoptive transfers of CD4+ CD25+ Tregs partially alleviate mouse premature ovarian insufficiency.

This study was designed to investigate the protective effect of CD4+ CD25+ regulatory T cells (Tregs) against zona pellucida glycoprotein 3 peptide (pZP3) immunization-induced premature ovarian insufficiency (POI) in mice. A mouse POI model was induced by two subcutaneous injections of pZP3 (50 nmol/L). Mice in the pZP3-Treg group were intraperitoneally injected with 5 × 105 CD4+ CD25+ Tregs after the POI model was established. Sex hormone levels, follicle numbers, apoptotic events, and the Akt/FOXO3a signaling pathway molecules in the ovaries were assessed. Compared with control group, the weight of ovaries in both pZP3 group and pZP3-Treg group was decreased and no difference was found between them. The number of follicles in the Treg transferred mice, like in pZP3 group, was significantly reduced compared to the control group, but showed a modest improvement when compared the pZP3 group alone. Significantly lower serum concentrations of follicle-stimulating hormone, luteinizing hormone, and anti-zona pellucida antibodies (AZPAbs) were found, while the concentrations of estradiol and anti-Mullerian hormone increased. In mechanism, Treg cell transfer to ZP3 treated mice restored the levels of Caspase3 to control levels, and partially restored Bax, however, had no effect on Bcl-2. Moreover, Treg cell transfer to ZP3 treated mice partially restored the levels of Akt and FOXO3a, and partially restored the ratios of p-Akt/Akt and p-FOXO3a/FOXO3a. In conclusion, Treg cells improved some aspects of ZP3-induced POI which may be mediate by suppressing ovarian cells apoptosis and involving the Akt/FOXO3a signaling pathway. Therefore, Treg cells may be protective against autoimmune POI.

RETRACTED ARTICLE: hPMSC transplantation restoring ovarian function in premature ovarian failure mice is associated with change of Th17/Tc17 and Th17/Treg cell ratios through the PI3K/Akt signal pathway

BackgroundHuman placenta-derived mesenchymal stem cell (hPMSC) transplantation has been demonstrated to be an effective way of recovering ovarian function in mice with autoimmune induced premature ovarian failure (POF). But the exact mechanism remains unclear. The goal of the present study is to investigate the role of immune factors (T-helper 17 (Th17), cytotoxic T (Tc17) and regulatory T (Treg) cells) in the recovery of ovarian function and whether the phosphatidylinositol 3-kinase (PI3K)/Akt signal pathway is involved in the regulation.MethodsThe inhibitor of PI3K/Akt was administered to observe its effect on ovarian function recovery and immune regulation. Serum levels of estradiol (E2), follicle stimulation hormone (FSH), luteinizing hormone (LH) and anti-Müllerian hormone (AMH)) and anti-Zona pellucida antibody (AZPAb) were measured by ELISA to evaluate ovarian function. The morphological changes of ovaries were observed by HE staining. Apoptosis of granular cells (GCs) was determined by detecting the expression of capase-3. Expression of p-Akt protein was detected by immunohistochemistry and western blot assay in ovarian tissues. The MTT assay was performed to assess GC proliferation. GC apoptosis was performed using flow cytometry analysis. Percentages of Th17, Tc17 and Treg cells were detected by flow cytometry. Expression of interleukin (IL)-17 in serum was measured by ELISA.ResultsLY294002 administration decreased serum levels of E2 and AMH, while the levels of FSH, LH and AZPAb in serum were increased compared with mice in the hPMSC transplantation group. The ovarian morphology presented as atrophy and fibrosis, with functional follicles exhausted. The expression of p-Akt in ovarian tissue was significantly decreased. Also, LY294002 administration significantly decreased proliferation and increased cell apoptosis in GCs, and for immune factors the ratios of Th17/Tc17 and Th17/Treg cells were significantly increased, as well as the serum levels of IL-17.ConclusionsOur data suggest that the PI3K/Akt signal pathway is involved in the recovery of ovarian function by changing the ratios of Th17/ Tc17 and Th17/Treg cells in POF mice following hPMSC transplantation.

RETRACTED ARTICLE: Effects of hPMSCs on granulosa cell apoptosis and AMH expression and their role in the restoration of ovary function in premature ovarian failure mice

BackgroundThis study was performed to determine the effects of human placenta mesenchymal stem cell (hPMSC) transplantation on granulosa cell apoptosis and anti-Müllerian hormone (AMH) and follicle-stimulating hormone receptor (FSHR) expression in autoimmune drug-induced premature ovarian failure (POF) mice. The aim of this research is to investigate the mechanisms of hPMSCs on ovarian reserve capacity.MethodsThe POF mice model was established by injection of zona pellucida 3 peptide (pZP3). hPMSC transplantation was conducted by intravenous injection into mice following pZP3 treatment. The follicle number was examined by histopathology. The serum levels of FSH, LH, E2, AMH and anti-zona pellucida antibody (AzpAb) were measured by enzyme-linked immunosorbent assay. AMH and FSHR expression in the ovary was analyzed by immunohistochemistry and western blot analysis. Granulosa cell apoptosis of the ovaries was examined by In Situ Cell Death Detection Kit. Granulosa cells were isolated and treated with SiAmh interference and hPMSC supernatant to observe the effects of AMH expression on granulosa cell apoptosis in vitro.ResultsThe results showed that hPMSC transplantation can significantly recover the estrus cycle in the POF group. Morphological staining showed that the basal follicles and sinus follicles after hPMSC transplantation were higher in POF mice than in those without treatment, and the follicle number was significantly decreased with atresia. The serum levels of FSH, LH and AzpAb in the hPMSC transplantation group were reduced considerably, but the E2 and AMH levels were significantly increased. After hPMSC transplantation, the AMH and FSHR expression in ovarian tissue was significantly higher than in the POF group as determined by immunochemistry and western blot analysis. The FSHR expression was shown in granulosa cells only, and FSHR expression increases with AMH expressed in the ovary; granulosa cell apoptosis was decreased following hPMSC transplantation. The same results were observed from the in-vitro study.ConclusionshPMSC transplantation can significantly improve the serum levels of high gonadotropin and low estrogen of POF mice, promote follicular development, inhibit excessive follicular atresia and granulosa cell apoptosis, and improve the ovarian reserve capacity. The mechanism may be achieved by increasing the expression of AMH and FSHR in ovaries.

Protective function of Bu Shen Huo Xue formula on the immunity of B6AF1 mice with experimental autoimmune premature ovarian failure.

Autoimmune abnormality is one of the main causes of premature ovarian failure (POF). Bu Shen Huo Xue formula (BSHXF), a traditional Chinese medicine formula, has been clinically used for the treatment of patients with POF in China. Regulatory T cells (Tregs) are important in the pathogenesis of autoimmune POF. The aim of the present study was to evaluate the immunoprotective effects of BSHXF on POF and the underlying mechanisms. An experimental autoimmune POF model was induced in B6AF1 mice with zona pellucida 3 (ZP3) fragments. Following modeling, BSHXF (31.53 g/kg/day) was orally administered for 4 weeks. CD4+ T lymphocytes, Tregs, anti-zona pellucida (anti-ZP) antibodies and cytokines were detected using flow cytometry, enzyme-linked immunosorbent assays, reverse transcription-quantitative polymerase chain reaction and immunohistochemistry. The results revealed that BSHXF exhibited an immunoprotective function and reduced inflammatory cell infiltration and damage to the ovary. BSHXF upregulated the percentage of CD4+ CD25+ forkhead box P3+ T cells in the spleen, effectively inhibiting the activation of CD4+ T lymphocytes. The proliferation of Tregs was increased in serum obtained from mice in the BSHXF group in vitro. Anti-ZP antibodies and interleukin-10 and interferon-γ levels were decreased in the serum in the BSHXF-treated group. The present study demonstrated that BSHXF had an immunoprotective effect on POF model mice. Additionally, it indicated that the protective mechanisms of BSHXF may be associated with an increase in Treg cells.

Role of Oxidative Stress and Chlamydia trachomatis in Infertile Women with Anti-Zona Pellucida Autoantibodies

Female infertility could be attributed to various factors including; sperm isoantobody, zona pellucid antibody, ovarian antibody, Chlamydia trachomatis infection and imbalance between oxygen radical and antioxygen radical. These factors might impair infertility by different mechanisms. A retrospective case-control study was performed on 105 infertile female including 75 (71.4%) with primary infertility and 30 (28.6%) with secondary infertility. Thirty healthy women were used as a control. Antisperm antibody (ASA), antizona pelucida antibody (AZP-Ab) and antiovarian antibody (AOA) were detected in the serum and cervical secretion using Enzyme labeled and Biotin labeled antibody. PCR technique was applied to detect the presence of Chlamydia trachomatis in the cervical secretion using the following primers: KL1-F and KL2-R, whereas IgG specific antibody was examined in the serum samples. According to infertility etiology, 29 (21.5%) were classified with unexplained infertility, 32 (23.7%) with polycystic ovary, 6 (4.4%) with endometriosis, 22 (16.3%) with tubal damage, 9 (6.7%) with hormonal imbalance, while other cases showed multiple causes of infertility. ASA were detected in the serum of infertile females (13.3%) and in (30%) of fertile one. Autoantibodies to zona pelucida were detected in the serum and the cervical secretion of infertile females (9.5% and 13.3%, respectively). No detectable levels of AZP-Ab were observed in fertile females. Statistically, the prevalence of ASA was significantly correlated to the AZP-Ab detected in the serum and cervical secretion of infertile females (P˃0.05). Aautoantibodies to different ovarian antigens were detected in the cervical secretion of 47 (44.8%) of the cases, in the absence of any detectable levels among fertile females. A high percentage of ASA, AZP-Ab and AOA were detected in primary infertility-related cases. However, with respect to the etiology of infertility, it seems that tubal damage, polycystic ovary and unexplained cases showed the highest percentages of both iso and autoantibodies. Serum samples obtained from fertile and infertile females were analyzed for the presence of C. trachomatis specific IgG. Five (4.76%) of etiologically tubal damage infertile females showed the presence of Chlamydial antibody with no detectable level in the serum of fertile females. To confirm the presence of C. trachomatis, PCR was used to identify Chlamydia DNA in the cervical secretion. Only positive serum samples revealed clear, sharp bands of amplified Chlamydial DNA with 241 bp. C. trachomatis was detected in infertile female with ASA (60%), serum and cervical secretion positive for AZP-Ab (60%, 80%, respectively) and in the cervical secretion positive for AOA (60%). A significant correlation was appeared between C. trachomatis infection and the presence of ASA and AZP-Ab (p˃0.05). Infertile females with Chlamydia infection has a high probability of inducing circulating and local ASA and AZP-Ab. Diagnosis of Chlamydia infection in the endocervix is recommended to assess their possible influence on ASA, AZP-Ab formation in infertile females.

Serum anti-Müllerian hormone dynamics in mares following immunocontraception with anti-zona pellucida or -GnRH vaccines

Circulating anti-Müllerian hormone concentration (AMH) is positively correlated to the number of small growing follicles in the mare and may reflect ovarian function. Dynamics of AMH during immunocontraception have not previously been investigated. This study aimed to compare serum AMH in mares following treatment with native porcine zona pellucida (pZP), recombinant pZP3 and pZP4 (reZP) or gonadotrophin releasing hormone (GnRH) vaccines, and saline-treated controls. Stored sera collected during two previous studies examining ovarian activity in mares during zona pellucida (ZP) or GnRH immunocontraception were analysed for serum AMH. Data were compared among treatment groups using mixed-effects linear regression and one-way ANOVA with post hoc testing. Correlations between AMH and previously reported clinical variables were estimated using Spearman's rho. Mares immunized against GnRH showed variable but detectable AMH throughout successive breeding and non-breeding seasons that were not significantly different to unvaccinated control mares. Mares treated with pZP demonstrated marked, reversible suppression of AMH. Mares immunized using reZP showed an intermediate effect. In the ZP study, AMH was positively correlated to serum progesterone concentrations, mean ovarian volumes and antral follicle counts, whereas no correlations between AMH and serum progesterone concentrations, mean ovarian volumes, or the presence of one or more follicles ≥20 mm in diameter were detected in the GnRH study. In conclusion, marked suppression of AMH during pZP immunocontraception, but not during GnRH immunocontraception, suggested enhanced suppression of ovarian follicular development and, or follicular function during pZP immunocontraception. Serum AMH concentrations may provide a novel tool for the assessment of ovarian function during ZP-based immunocontraception.

The association of different auto-antibodies against ovarian tissues and gonadotropins and poor ovarian response in intracytoplasmic sperm injection cycles

This study aimed to assess the possible association between ovarian auto-antibodies and poor ovarian response to controlled ovarian hyperstimulation (COH) in patients undergoing intracytoplasmic sperm injection (ICSI) cycles. In total, 42 poor responders and 43 male factor subjects were enrolled in the study and underwent either a standard long gonadotropin-releasing hormone (GnRH) agonist or antagonist protocol. Anti-ovarian, anti-oocyte, anti-zona pellucida (anti-ZP) and anti-gonadotropin antibodies in their sera and follicular fluid (FF) were measured by an enzyme-linked immunosorbent assay technique (ELISA). The mean follicular fluid anti-oocyte antibody [ratio of optical density (OD) sample/OD Control] was significantly higher in poor responders compared to the normal group (2.40 ± 1.55 versus 1.72 ± 0.71, p = 0.012). The linear regression analysis showed an inverse correlation between FF anti-oocyte antibody concentrations and the number of: (i) retrieved oocytes (B = −1.212, r = −0.235, p = 0.030); (ii) mature oocytes (B = −1.042, r = −0.234, p = 0.031); (iii) embryos available (B = −0.713, r = −0.228, p = 0.036); and (iv) good quality embryos (B = −0.369, r = −0.229, p = 0.035). However, there were no significant differences between two groups in terms of FF and serum anti-ovarian, anti-gonadotropins and anti-ZP antibodies. The Pearson correlation analysis on 85 infertile patients showed a positive correlation between age and the levels of FF anti-oocyte antibody (r = 0.276, p = 0.010). This study demonstrated that FF anti-oocyte antibody could be associated with poor response to COH in ICSI cycles.

The role of genetic and autoimmune factors in poor ovarian response in IVF programs

Background: The poor ovarian response (POR) to ovarian stimulation significantly decreases the efficiency of assisted reproduction technologies. The genesis of POR is not well understood, and can result from various genetic and immune factors.Purpose: To determine the role of genetic (number of CGG repeats in the FMR1-gene) and autoimmune factors (anti-ovarian antibodies) in POR in IVF programs.Methods: In a cross-sectional study, 150 infertile women were divided into the POR group (n = 65) and the group with a normal ovarian response (n = 85). All the patients underwent the detection of CGG repeats in the FMR1-gene by polymerase chain reaction, serum anti-zona pellucida antibodies (AZPA) and anti-ovarian antibodies by enzyme-linked immunosorbent assay.Results: The risk of POR was higher in patients with CGG counts > 34, but not in the patients with CGG counts < 26. The higher level of AZPA was found in the patients with the 26–34 CGG repeats compared to the patients with <26/> 34 CGG repeats.Conclusion: A higher number of CGG repeats in the FMR1-gene may be a risk factor of POR in IVF programs. Normal CGG counts in regard to ovarian reserve (26–34) suggest the autoimmune-driven POR.

Incidence of antibodies in women after failure of assisted reproduction

The aim of this study is to determine to what degree antisperm allo-antibodies (ASA), anti-ovarian antibodies (AOA), anti-zona pellucida antibodies (AZPA) and seven anti-phospholipid antibodies (APLA) can explain the failure of assisted reproduction technology (ART) in women. Among the causes of reproductive failure are allo- and autoimmune reactions of the organism against reproductive tissues and cells. We examined a sample of 43 selected women aged 27 to 45 after failure of assisted reproduction technology (ART) via intrauterine insemination (IUI), in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Sera from these women were tested by ELISA (enzyme-linked immunosorbent assay) (total Ig) for the presence of anti-sperm allo-antibodies (ASA), anti-ovarian antibodies (AOA), anti-zona pellucida antibodies (AZPA) and seven anti-phospholipid antibodies (APLA). We found 22 (51 %) patients positive for the examined antibodies. Among patients with positive results were 3 miscarriages, 3 biochemical pregnancies (↑ hCG) and 16 with no evidence of conception. Positive APLA was found in 17 patients. Selected immunological parameters were negative in 21 (49 %) patients, which means that the tested allo- and auto-antibodies did not cause the failure of ART. This demonstrates that the range of diagnostic tests limits the success of diagnosis and the method of dealing with infertile couples. The use of cardiolipin as a screening test appears to be insufficient, because in our sample it would have caused a 35 % error rate. Investigation of antibodies against seven phospholipids is thus considered to be reasonable and is beneficial for infertility diagnosis (Tab. 2, Ref. 36).

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. According to the levels of autoantibodies we have to choose preventive treatment to protect mother and her foetus.

Comparative study of different therapeutic methods on autoimmune

To investigate the effects of different methods of treatment, including tonifying the kidney, activating blood and soothing the liver, on mice with autoimmune premature ovarian failure (POF). After preparing ovarian antigen with the ovarian tissue of female BALB/C mice, POF in female BALB/C mice was induced by immunization injection of the ovarian antigen on multiple subcutaneous sites and two posterior soles. The mice were treated with Liuwei Dihuang Pill, Shaofu Zhuyu Granule, Chaihu Shugan Pill respectively after the third immunization. Levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E(2)), prolactin (PRL), testosterone (T), anti-ovarian antibodies (AOAb) and anti-zona pellucida antibody (aZP) in sera were analyzed by enzyme-linked immunosorbent assay. Compared with the normal group, the levels of serum FSH and LH were increased and the level of E(2) was decreased in the untreated group. AOAb and aZP positive rates in the untreated group were obviously higher than those in the normal group. Liuwei Dihuang Pill could significantly reduce the levels of FSH and LH and increase the level of E(2) as compared with the untreated group, but there were no significant differences in the AOAb and aZP positive rates. Shaofu Zhuyu Granule and Chaihu Shugan Pill could significantly reduce the positive rates of AOAb and aZP as compared with the untreated group, but no significant effect on the levels of FSH, LH and E(2) were observed. The drugs had no effects on serum contents of PRL and T. Tonifying the kidney method can recover the ovarian function in POF mice mainly by regulating the hormone levels. Activating blood and soothing the liver methods can improve the humoral immune function in POF mice.

Harmful effects of anti-zona pellucida antibodies in folliculogenesis, oogenesis, and fertilization

The zona pellucida (ZP) is an extracellular matrix that surrounds the mammalian oocyte and plays an important role in normal folliculogenesis and fertilization. Because of its strong immunogenicity and its possible relation with premature ovarian failure, we conducted the present study to examine whether or not anti-ZP antibodies impaired folliculogenesis. Mouse preantral follicles were cultured with anti-ZP antibodies to evaluate the effects on follicle growth and antral formation. The cultured follicles were also examined by electron microscope and assessed for oocyte maturation, fertilization capacity, and embryo development. The results showed that follicles cultured with anti-ZP antibodies had a smaller diameter than the controls. Also, these antibodies reduced antral formation, mucification, maturation of oocytes (metaphase II), and fertilization rates. Morphologically, ZP thickness was lower in the anti-ZP antibody groups. The quantity of granulosa cell microvillous processes that transverse the ZP was diminished in follicles cultured with anti-ZP antibodies. In conclusion, anti-ZP antibodies were harmful to the normal development of mouse follicles and oocytes in vitro. These antibodies may be a cause of premature ovarian failure syndrome because they disrupt the gap junctions between the oocyte and granulosa cells and, as a consequence, damage the bidirectional communication necessary for normal folliculogenesis.

Effects of immunocontraception on behavior in fallow deer (Dama dama)

Immunocontraceptives can control growth in wild and captive populations; however, in doing so, they should not disrupt species-typical behavior patterns. The presence of treated females could disrupt social interactions in a population; yet, few studies have examined effects of immunocontraception on behavior. The goal of this project was to determine whether behavior in a fallow deer (Dama dama) population vaccinated with porcine anti-zona pellucida (pZP)-immunocontraceptives differed from behavior in a population of unvaccinated deer. We predicted that pZP-treated females would spend a higher percentage of time exhibiting mating and dominance behavior than nontreated females, and that males interacting with pZP-treated females would spend a higher percentage of time demonstrating mating and aggressive behavior than males interacting with nontreated females. We recorded activity budgets of males and females in two fallow deer populations, one pZP-treated and the other untreated, before, during, and after rut. Females did not differ in time spent displaying mating or dominance behavior compared to nontreated females. Males coexisting with nontreated females spent more time exhibiting aggressive behavior during rut than males living with pZP-treated females, but males did not differ in time spent in mating behavior. Thus, immunocontraception did not seem to affect behavior adversely. However, sample sizes, living conditions, and sex ratios may have affected the results. Given these limitations, future research is needed to confirm our findings. Zoo Biol 27:49-61, 2008. (c) 2007 Wiley-Liss, Inc.

Detection of antizona pellucida antibodies in the sera from premature ovarian failure patients by a highly specific test

To develop a highly specific test for the detection of antizona pellucida (ZP) antibodies in the sera from premature ovarian failure (POF) patients. Laboratory study. University hospital. Twenty-seven idiopathic POF patients, 30 control women, and 30 healthy males. Anti-ZP antibodies were detected by the microdot assay using a very small amount of human ZP or porcine ZP. The effect of anti-ZP antibodies on sperm-ZP binding was examined by hemizona assay. Results from the microdot assay and hemizona assay. By the microdot assay using human ZP, the sera from POF patients reacted significantly stronger than those of control women and healthy males. However, no obvious difference could be found by the same assay using porcine ZP among these three groups. Anti-ZP antibodies against sera from some POF patients showed significant blocking effects on sperm-ZP binding assessed by hemizona assay. Anti-ZP antibodies were detected in 7 of 27 POF patients, while none were detected in control women and healthy males. Some idiopathic POF patients have anti-ZP antibodies in their sera, which were detected with high specificity by a newly developed microdot assay using a very small amount of human ZP.

Quelles cibles antigéniques dans l’ovaire ?

The ovary can be the target of an autoimmune disease involving many different autoantigens. The clinical feature of this disease often results in premature ovarian failure or infertility and may be either isolated or associated with other autoimmune pathologies, especially with adrenal autoimmunity. The diagnosis of an autoimmune mechanism relies on the presence of anti-ovarian antibodies, whose prevalence is quite variable according to the different methods used to detect them, and to the different stages of the disease. In addition, their clinical significance is not always clear, as to their pathologic or epiphenomenal nature. However, the study of these autoantibodies has led to the identification of some of their antigenic targets which have to be known for a better understanding of the pathologic mechanisms involved. This paper reviews anti-steroid producing cells, anti-gonadotrophin receptor, anti-gonadotrophin, anti-corpus luteum, anti-zona pellucida and anti-oocyte antibodies.

Endometriosis in reproductive immunology.

Endometriosis is suggested to represent an autoimmune disorder, but what is the prevalence of autoantibodies to antigens relevant to reproduction? 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. Endometriosis I-II were associated with higher serum and peritoneal fluid levels of aPLs against inositol, cardiolipin, ethanolamine, and beta2-glycoprotein I. Forty percent of patients were positive for aZPA. Patients with lesions of endometriosis stage I-II had more autoantibodies than those with stage III-IV, and may be immunologically more active. This result may be significant for future treatments such as in vitro fertilization and embryo transfer.