Akane Kondo

Increased natural Killer-Cell activity is associated with infertile women

PROBLEM: An increase in natural killer (NK)‐cell activity has been observed in women with unexplained recurrent miscarriages. Because of the many similarities between infertility and early pregnancy loss patients, we investigated whether infertile women had raised NK‐cell activity.
 METHOD OF STUDY: We tested 94 infertile women who, in spite of treatment, were unable to conceive for 6 or more months. NK‐cell activity was measured by using a chromium‐51 release cytotoxicity assay, with K562 human myeloid leukemia cells as targets.
 RESULTS: NK‐cell activity of the infertile group (mean±SD; 40.2%±14.7) was significantly higher than the control group (31.5%±11.9, P<0.0001). The increased NK‐cell activity was not associated with age, infertile duration, depression scores, treated hyperprolactinemia, or treated endometriosis.
 CONCLUSIONS: In certain patients, elevated NK‐cell activity may be considered an independent risk factor for infertility.

Different antiphospholipid antibody specificities are found in association with early repeated pregnancy loss versus recurrent IVF-failure Patients

PROBLEM: Patients having in vitro fertilization and embryo transfer (IVF‐ET) failures show an increased incidence of antiphospholipid (aPL) antibodies; but controversy exists whether aPL can induce IVF‐failure. This study was designed to compare aPL specificities between recurrent IVF‐failure patients versus repeated early pregnancy loss (RPL) patients.
 METHOD OF STUDY: Anticardiolipin (aCL), lupus anticoagulant (LA), antiphosphatidylserine (aPS), antiphosphatidylethanolamine (aPE), and antinuclear antibodies (ANA) were measured in 74 recurrent IVF‐ET failure patients and compared with 273 early RPL patients (<10 weeks).
 RESULTS: An increased incidence of IgG‐aPE and ANA was observed for both groups in comparison with controls. Patients with recurrent IVF‐ET failure showed a significantly higher prevalence of IgG‐aPS (P=0.02) and IgG‐aCL (P=0.02) when compared with early RPL patients or controls.
 CONCLUSIONS: IgG‐aPS and IgG‐aCL may be responsible for some IVF‐failures. Additional studies are needed to clarify the pathogenic role of IgG‐aPS and IgG‐aCL on IVF‐ET failure.

Pregnancy Loss and Endometriosis: Pathogenic Role of Anti‐Laminin‐1 Autoantibodies

Abstract: Laminin‐1 is a major multifunctional glycoprotein that forms an integral part of the scaffolding network of basement membranes, and is the earliest synthesized component during embryogenesis. This protein (α1β1γ1) plays an important role in basement membrane assembly and epiblast differentiation during embryonic development. Anti‐laminin‐1 autoantibodies are known to cause infertility and recurrent spontaneous abortion in animals. Recently, we reported that the presence of IgG anti‐laminin‐1 antibodies (Abs) in the blood is significantly associated with recurrent first‐trimester miscarriages and subsequent negative pregnancy outcomes. Interestingly, these antibodies are also strongly associated with infertility, especially infertility caused by endometriosis. Laminin‐α1, laminin‐β1, and laminin‐γ1 mRNAs were also detected in 90% of endometriotic lesions, and all laminin‐α1, laminin‐β1, and laminin‐γ1 chains were localized to the basement membranes of glandular epithelium in endometriotic peritoneal lesions. ELISA showed specific reactivity of the autoantibodies to a particular region of the laminin‐1 molecule, that is, the α1 chain G domain. IgM monoclonal anti‐laminin‐1 Abs, which we recently established, also recognized the G domain and cross‐reacted with human α1 chain located in the basement membrane of the glandular epithelium of human endometrium. We also established an animal model that produced high titers of anti‐laminin‐1 Abs after immunization with mouse laminin‐1. Anti‐laminin‐1 Abs from the immunized mice caused a higher fetal resorption rate with lower embryonic and placental weights. Thus, anti‐laminin‐1 Abs may be important in the development of autoimmune‐mediated reproductive failures, and the assessment of the such antibodies may provide a novel means for noninvasive diagnosis of endometriosis.

Anti-annexin A5 Antibodies in Reproductive Failures in Relation to Antiphospholipid Antibodies and Phosphatidylserine

Problem: The presence of IgG anti‐annexin A5 (IgGαA5) and/or antiphospholipid antibodies (aPL) are risk factors associated with recurrent spontaneous abortion. Problems are whether IgA anti‐annexin A5 (IgAαA5) is pathogenic, and how IgGαA5 works.

Preconception Peripheral Natural Killer Cell Activity as a Predictor of Pregnancy Outcome in Patients with Unexplained Infertility

Problem:  Preconception high peripheral natural killer (NK) cell activity in women with recurrent spontaneous abortion can predict subsequent miscarriages. We have examined prospectively, for the first time, the pregnancy rate in patients with unexplained infertility by measuring the peripheral NK activity.

IgG‐Antiphospholipid Antibodies in Follicular Fluid of IVF‐ET Patients are Related to Low Fertilization Rate of Their Oocytes

Patients undergoing in vitro fertilization and embryo transfer (IVF‐ET) failures show an increased incidence of antiphospholipid antibodies (aPL) in their blood. The physiological manifestations of aPL in this patient group are nonetheless controversial. Pathological effects of aPL on embryos in vitro have been documented. We questioned whether aPL if found in follicular fluids (FFs) could result in embryonic damage.

Anti-laminin-1 autoantibodies, pregnancy loss and endometriosis

Laminin-1 is a major component and multifunctional glycoprotein of basement membranes that consists of three different subunits, α1, β1 and γ1 chains. It is the earliest synthesized network-forming protein during embryogenesis and plays an important role in embryonic development, embryonic implantation and placentation. We have recently shown that IgG anti-laminin-1 antibodies were significantly associated with recurrent first-trimester miscarriages and with subsequent pregnancy outcome. Interestingly, these antibodies were also observed in patients with endometriosis-associated infertility but not in patients with other causes of infertility, including tubal factors, hormonal and uterine abnormalities. Laminin-α1, -β1 and -γ1 mRNAs have been detected in 90% of endometriotic lesions and all laminin-α1, -β1 and -γ1 chains were localized in the basement membranes of glandular epithelium in endometriotic peritoneal lesions. Western blot analysis showed that anti-laminin-1 antibodies from those patients reacted with all laminin-1s chains. ELISA also confirmed that one of the target epitopes for these antibodies was located in a particular region of the laminin-1 molecule, i.e. the carboxyl-terminal globular G domain of α1 chain. IgM monoclonal anti-laminin-1 autoantibody, that we recently established, also recognized the G domain. Anti-laminin-1 antibodies from mice immunized with –mouse— laminin-1, caused a higher fetal resorption rate with lower embryonic and placental weights. Thus, anti-laminin-1 antibodies may be important in development of autoimmune-mediated reproductive failures and the assessment of the antibodies may provide a novel non-invasive diagnosis of endometriosis.

Characterization of a murine anti-laminin-1 monoclonal antibody (AK8) produced by immunization with mouse-derived laminin-1

Laminin-1 is a structural glycoprotein that forms an integral part of the scaffolding of basement membranes, and plays an important role during embryonic development. We have recently demonstrated a significant association between anti-laminin-1 antibodies (Abs) and reproductive failure, such as recurrent spontaneous abortions and infertility-associated endometriosis in both human and mouse studies. In the present study, we established an IgM (μ,κ) monoclonal anti-laminin-1 Ab (AK8) by immunizing mice with mouse Engelbreth-Holm-Swarm sarcoma (EHS)-derived laminin-α1. The AK8 monoclonal antibody (mAb) reacted with particular peptide sequences from the globular G domain of mouse laminin-α1 chain of using ELISA and Western blot techniques. The peptide tertiary structure of the epitope recognized by AK8 mAb was predicted using eight synthesized domain peptide sequences and three consensus sequences obtained by phage displayed random peptide library. Basement membranes of endometrium of pregnant mice and humans were immunostained with AK8 mAb. Thus, AK8 mAb recognized a common structure present in the G domain of the laminin-1 chain in both mice and humans. The passive immunization of mice with AK8 mAb may represent a suitable animal model for anti-laminin-1 Ab-mediated reproductive failure.

Noninvasive measurement of fetal augmentation index by fetal aortic diameter pulse and flow velocity waveforms

Objective. To study fetal systemic arterial stiffness in normal fetuses and compromised fetuses who had umbilical placental insufficiency (UPI). Design. Prospective study. Setting. University departments. Sample. A total of 118 normal fetuses (21–40 weeks) and 55 fetuses (UPI group) with evidence of potential compromise (high umbilical artery pulsatility index). Methods. A new real‐time noninvasive measurement system based on a combined Doppler ultrasound and echo‐tracking system was used as a measure of aortic/systemic arterial stiffness. The augmentation index (AI) of the fetal thoracic descending aorta was measured by using simultaneous measurements of diameter pulse and flow velocity waveforms. Main Outcome Measure. Augmentation index as a measure of stiffness. Results. In normal fetuses, successful measurements for obtaining the AI were achieved in 103 of 118 fetuses. In the normal group, the AI, as well as placental resistance, decreased during the second trimester; in contrast, an increase in the AI was observed during the third trimester. Using the AI values from the normal group, the UPI group was divided into two subgroups: 29 fetuses with a normal AI and 26 fetuses with a high AI. The clinical outcome was significantly worse in the latter subgroup compared with the normal subgroup. Conclusions. The increase of afterload caused by a high umbilical placental resistance was associated with a decrease of aortic distensibility in the compromised fetuses, suggesting an alteration of aortic wall structure.

Exome and copy number variation analyses of Mayer–Rokitansky–Küster– Hauser syndrome

Mayer–Rokitansky–Küster–Hauser (MRKH) syndrome is characterized by congenital absence of the vagina and uterus. We conducted genome-wide SNP analyses and exome sequencing to detect the causes of MRKH syndrome. We identified de novo variants of MYCBP2, NAV3, and PTPN3 in three families and a variant of MYCBP2 in a sporadic case. Here, we demonstrated the partial genetic makeup of Japanese MRKH syndrome.