Tsunehisa Makino

Flow-cytometric analysis of immune cell populations in human decidua from various types of first-trimester pregnancy

We undertook an investigation in which flow cytometry was used to characterize immune cell populations in the decidua of first-trimester normal pregnancies, spontaneous abortions, and ectopic pregnancies in comparison to the nonpregnant endometrium to demonstrate how the proportions of immunocompetent cell populations at the fetomaternal interface are influenced by the presence and state of a fetoplacental allograft. No significant differences were found in the decidua of the different types of first-trimester pregnancy in the proportions of the CD45+, CD14+, CD3+, CD4+, CD8+, CD19+, CD3-/CD16+ and/or CD56+, CD3+/CD16+ and/or CD56+, CD4+/Leu-8+, CD4+/Leu-8-, CD8+/CD11b+, CD8+/CD11b-, and CD3+/HLA-DR- decidual leukocyte subsets. However, the percentage of decidual CD3+/HLA-DR+ cells, which are characteristic of activated T cells, was significantly higher in spontaneous abortions than in normal pregnancies (p less than 0.05). This suggests that the accumulation of decidual leukocytes may be regulated mainly by hormones and/or cytokines rather than by the presence and state of an intrauterine conceptus and that on/off-switching of activation of decidual T cells may be associated with successful maintenance of the implanted embryo.

Survey of 1120 Japanese women with a history of recurrent spontaneous abortions

Our recent 7-year clinical survey showed that among the 1120 women with repeated spontaneous abortions registered in this clinic, 2898 out of a total of 3216 pregnancies (90.1%) had terminated in spontaneous abortion. Among these wastages, 84.2% occurred before 12 weeks of gestation, and 11.1 percent occurred between 12 and 15 weeks. Through routine examination of reproductive wastage, 82 (9.9%) of the 825 Japanese couples examined were shown to have either a chromosomal abnormality or normal variants in the wife and/or husband, thus demonstrating no racial difference in the incidence of chromosomal abnormalities in infertile patients in comparison with studies performed in other countries. One hundred and forty-seven congenital uterine anomalies (14.7%) were found in 1000 hysterosalpingographies, and 12 of 148 examined females were positive for anti-cardiolipin antibody. 393 other females with no major abnormalities likely to induce spontaneous abortions were indicated for immunotherapy. Ample time spent on genetic counseling prevented further reproductive wastage, and ideal metroplasty resulted in a successful post-operative pregnancy course in more than 85% of cases. Immunosuppressant and anticoagulant therapy decreased the serum titer of anti-cardiolipin antibody, enabling pregnancies to be maintained to term. Immunotherapy utilizing the husbands lymphocytes also brought more than 80% of pregnancies to successful completion, with 200 deliveries achieved with this therapy. In contrast, 64.1% of pregnancies again terminated spontaneously in patients who were indicated for immunotherapy but did not receive treatment. The findings of the present study suggest that the causes of reproductive wastage, especially the etiology of early recurrent spontaneous abortion, are complex.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of Narcotic Analgesics and Naloxone on Proestrous Surges of LH, FSH and Prolactin in Rats

Morphine (50 mg/kg), given at 12:00 h and 14:00 h to proestrous rats, completely inhibited the proestrous surges of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (Prl). However, when it was given at 16:00 h, the inhibitory effect of morphine (Mor) was decreased. Pethidine and nalorphine inhibited the proestrous surges, while pentazocine showed a weaker inhibitory effect. Naloxone (Nx), given simultaneously or 2 h after morphine (Mor), at 1/20 the dose of Mor, inhibited the blockade of the proestrous gonadotropin release exerted by Mor, while the onset of the surges was delayed when it was given 3 h after Mor.

A possible mechanism of immunotherapy for patients with recurrent spontaneous abortions

ABSTRACT: The mechanism of the beneficial effect of immunotherapy for human reproductive wastage remains to be elucidated. Induction of blocking antibodies such as anti‐HLA class II antibodies and anti‐idiotypic antibodies was investigated as the mechanism of specific immunosuppression in pregnancy. We reported the changes in the mixed lymphocyte reaction (MLR), T‐cell subsets, and generation of anti‐idiotypic antibodies after immunotherapy compared to before immunotherapy. MLR was significantly (P < 0.001) inhibited after the immunization. The mean inhibition rate was 50.2%, suggesting that MLR blocking antibodies were induced by immunotherapy. Binding of autoantibodies to alloactivated maternal lymphoblasts against the paternal lymphocytes was detected in postimmunization cases in two‐color flow‐cytometric experiments. This suggests that anti‐idiotypic antibodies were induced by the immunotherapy. The percentage of cytotoxic T‐cells was significantly decreased (P < 0.05) and the percentage of suppressor T‐cells was significantly increased (P < 0.01) after the immunotherapy, suggesting that a cell‐mediated immune response was induced by the immunotherapy.

Flow cytometric crossmatch and early pregnancy loss in women with a history of recurrent spontaneous abortions who underwent paternal leukocyte immunotherapy

OBJECTIVEnThe purpose of our study was to evaluate the clinical significance of flow cytometric crossmatch in managing women with unexplained recurrent spontaneous abortions who have been immunized with paternal leukocytes.nnnSTUDY DESIGNnTwo-color flow cytometric crossmatch was performed after immunotherapy in 69 women with recurrent spontaneous abortions to detect maternal immunoglobulin G antibodies against paternal T lymphocytes. A positive flow cytometric crossmatch was determined by a > or = 20 channel increases (256 channel linear scale-log amplification) in the fluorescence intensity of the T-cell peak. We analyzed the clinical correlation between flow cytometric crossmatch results and subsequent pregnancy outcome in those patients using the Fisher exact test.nnnRESULTSnIn 46 subsequent pregnancies among 69 immunized women the incidence of early subsequent pregnancy wastage was only 6 (17.1%) of 35 in flow cytometric crossmatch-positive patients and 7 (63.6%) of 11 in flow cytometric crossmatch-negative patients (p < 0.01).nnnCONCLUSIONSnThese data indicate that a two-color T cell flow cytometric crossmatch predicts subsequent pregnancy outcome in patients undergoing immunotherapy for recurrent spontaneous abortion.

Effect of prostaglandin D2 on gonadotropin release from rat anterior pituitary in vitro

A new prostaglandin D2 (PGD2), which is highly concentrated in the rat hypothalamic-pituitary system, was investigated for its gonadotropin release and synthesis by radioimmunoassay (RIA). Significant increases in the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were observed when the anterior pituitaries were incubated for 4 hours in vitro in the presence of 500 micrograms PGD2/ml incubation medium. Little change in LH and FSH contents in the rat anterior pituitary was observed between the control and PGD2-stimulated groups during incubation. In addition, more than 200 micrograms PGD2/ml induced a marked increase of cyclic adenosine 3,5 monophosphate (cAMP) formation in the rat anterior pituitary, and this was observed within 10 minutes after stimulation by PGD2. These data indicate that PGD2 directly induces a marked release of both LH and FSH from the gonadotrophs, and the mechanism of action of PGD2 could be mediated by cAMP formation.

Influence of Immunotherapy on Antisperm Antibody Titer in Unexplained Recurrent Aborters

PROBLEM: The mechanism of the beneficial effect of immunotherapy for human reproductive wastage remains to be elucidated. Because some women with unexplained recurrent spontaneous abortion are immunized with their partners lymphocytes, it is important to determine whether such immunization results in elevation or enhancement of immunity to spermatozoa, because antigenic cross‐reactivity between lymphocytes and spermatozoa has been reported.

Leukocyte Subpopulation Changes in Rats With Autotransplanted Endometrium and the Effect of Danazol

PROBLEM: This study examines immune cell populations in rats with autotransplanted endometrium and determines the effect of danazol on leukocyte subsets.

Current comprehensive therapy of habitual abortion

Habitual abortion is sometimes an incurable pathogenetic state. Even more serious to both the patient and gynecologist is that the incidence of repeated reproductive wastage is higher in subsequent pregnancies. We registered more than 1,000 women with repeated spontaneous abortions in this clinic for 5 years and analyzed their pathogenesis for the wastages by several different approaches. One hundred twenty-four women were diagnosed as having a congenital uterine abnormality. Metroplasty was performed in 50 patients, resulting in 82% successful subsequent pregnancies, whereas more than 98% of the pregnancies had terminated in spontaneous abortion before the operation. By measuring anticardiolipin antibody (ACA) and lupus anticoagulant (LA), the present study diagnosed several cases as antiphospholipid syndrome, and successfully pregnant patients were observed and treated with appropriate medications. Fifty-five partners of 54 couples had either a chromosomal abnormality or normal variants, showing a high incidence of spontaneous abortion in each type of abnormality. In the other 311 cases, female partners were treated by the immunotherapy of the husbands lymphocytes in pre- and postgestational periods. Among them, 200 women became pregnant, and 147 cases successfully maintained the pregnancy. These studies indicate that early diagnosis through comprehensive examinations is essential for effective treatment of curable fetal loss in human reproduction.

A Study on the Biological Action of Hypothalamic Tridecapeptide, Neurotensin

Investigations on various kinds of biological actions of a newly purified hypothalamic tridecapeptide, neurotensin, were performed both in vivo and in vitro by utilizing experimental animal models. The effect of neurotensin on pituitary gonadotropin release was studied in ovariectomized estrogen-progesterone-treated rats by the measurement of serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin (PRL) in radioimmunoassays. Neurotensin (340 mg/100 g BW) significantly increased serum LH and FSH 30 minutes after an intravenous injection (p smaller than 0.05) and lowered serum prolactin concentrations significantly (p less than 0.025). Bradykinin and substance P showed on significant effect on serum LH and FSH release. Intraperitoneal or intravenous administration of 0.5 n.mole neurotensin lowered blood pressure in intact mature rats from the range of 90 approximately 100 mmHg to 50 approximately 60 mmHg; however, tachyphylaxis was observed after repeated injections of the same dose of this peptide. Neurotensin was as potent as bradykinin in inducing rat duodenum relaxation and guinea pig ileum contraction in vitro. Theses effects on neurotensin and bradykinin on the intestines were not inhibited by the pre-treatment of phentolamine, propranolol, methysergide and pyribenzamine. Bradykinin induced contractions of the uterus in proestrous rats, but neurotensin induced no marked contraction. These results suggest that neurotensin in not hypothalamic releasing or inhibiting factor but possess the nature of kinin.