Koji Kanazawa

Cytotoxic antibody against trophoblast and lymphocytes present in pregnancy with intra- uterine fetal growth retardation and its relation to anti-phospholipid antibody

To elucidate immunological mechanisms involved in the genesis of intra-uterine fetal growth retardation (IUGR), an in vitro cytotoxicity assay against normal trophoblast and lymphocytes was performed. The data demonstrated the existence of cytotoxic antibody directed against trophoblast exclusively in the IgG fraction of the sera of 9 out of 15 mothers with IUGR, but in none of the sera from normal pregnant women. This antibody showed differential reactivity patterns that may be indicative women. This antibody showed differential reactivity patterns that may be trophoblast in common. Out of 9 IUGR mothers with this cytotoxic antibody, in 6 cases (66.7%) chronic villitis was evident upon microscopic examination of the placenta, the frequency being significantly higher than that in IUGR mothers without cytotoxicity or in control mothers (P less than 0.02). It is suggested that in situ inflammatory change triggered by this antibody might lead to IUGR through chronic villitis. This antibody showed cross-reactivity with anti-negatively charged phospholipid antibody, as confirmed by an absorption experiment, indicating that the trophoblast antigenic stimuli in pregnancy can induce the production of various autoantibodies including anti-phospholipid antibodies. These results are of interest in relation to the pathogenesis of autoimmune diseases.

Result of Immunotherapy on Patients With Unexplained Recurrent Abortion: A Beneficial Treatment for Patients With Negative Blocking Antibodies

ABSTRACT: Thirty‐nine unexplained recurrent aborters underwent vaccination using husbands lymphocytes according to the previously reported protocol. No mixed lymphocyte culture reaction‐blocking antibodies (MLR‐BAbs) were observed in these patients prior to vaccination. Of 35 newly pregnant patients after vaccination(s), pregnancy successfully continued in 28 (80.0%) and have already been terminated with a liveborn offspring. Pregnancy outcome was also analyzed in unexplained recurrent aborters who revealed positive MLR‐BAbs without immunotherapy. In this group, out of eight pregnancies in seven patients, five (62.5%) continued beyond their critical period of 14 wks of gestation. Three infants born from these pregnancies, however, presented severe abnormalities. Furthermore, outcome of 14 pregnancies in 12 unexplained recurrent aborters with negative MLR‐BAbs was analyzed since they had become pregnant without immunotherapy; pregnancy was successfully continued in only four cases (28.6%). Thus, vaccination using husbands lymphocytes on unexplained recurrent aborters with negative MLR‐BAbs is suggested to be effective. In addition, it is suggested that immunotherapy for patients with positive MLR‐BAbs should be carefully followed.

Immunohistochemical localization of HLA antigens and placental proteins (αhCG, βhCG CTP, hPL and SP1 in villous and extravillous trophoblast in normal human pregnancy: a distinctive pathway of differentiation of extravillous trophoblast

Abstract Immunohistochemical localization of HLA antigens and plancental proteins (αhCG, βhCG CTP, hPL and SP 1 ) in villous and extravillous trophoblast at various stages of normal human gestation were studied, using hysterectomy specimens. In the chorionic villi, the capacity for synthesizing placental proteins seemed to develop in parallel with the morphological change from mononuclear cells to multinucleated syncytiotrophoblast and no villous trophoblast expressed HLA antigens. In contrast, extravillous trophoblast, including the multinucleated trophoblastic cells at the deciduomuscular junction, expressed HLA-A, -B, and -C, and their capacity for synthesizing placental proteins did not seem to correspond with the degree of morphological change: the location of αhCG, βhCG CTP and SP 1 was restricted to mononuclear trophoblast in the superficial decidua, while hPL was present extensively in extravillous trophoblast. These findings strongly suggest that extravillous trophoblast possesses many distinctive biological features and differentiates in an independent manner. Mononuclear trophoblast forming the cell columns was also positive for HLA-A, -B, and -C, and no placental protein was demonstrated in these cells; this, together with previous morphological observations, may indicate the germinative nature of these cells.

Reactivity of two monoclonal antibodies (Troma 1 and CAM 5.2) on human tissue sections: analysis of their usefulness as a histological trophoblast marker in normal pregnancy and trophoblastic disease

In normal and molar pregnancy, a morphological discrimination between nonvillous trophoblasts which lie scattered in the placental bed and surrounding maternal cells is considered to be difficult. We examined the reactivity of two monoclonal antibodies (Troma 1 and CAM 5.2) against cytokeratin by an immunoperoxidase technique and analyzed their usefulness as a histological trophoblast marker. Materials were taken from 42 uteri with normal pregnancy, 7 uteri with hydatidiform mole, 2 uteri with gestational choriocarcinoma, 1 fallopian tube with nongestational choriocarcinoma, 5 delivered term placentae of normal pregnancy, and 5 nongestational uteri. The reactivities of Troma 1 on frozen sections and those of CAM 5.2 on paraffin sections were identical. They reacted with surface epithelium and gland epithelium in the nongestational uterine corpus. In the implantation site of normal and molar pregnancy, they reacted with villous and nonvillous trophoblasts as well as endometrial gland epithelium. In gestational and nongestational choriocarcinoma, they reacted with carcinoma cells specifically. Since the histological detection of gland epithelium may not be difficult, it was concluded that the two antibodies were very beneficial as a histological marker for trophoblasts in normal pregnancy and trophoblastic disease.

Is Immunotherapy for Habitual Aborters an Immunologically Hazardous Procedure for Infants

ABSTRACT: Physical development and tests of immunologic function are reported from the first year of life for 13 infants born to mothers who were habitual aborters and who had undergone subcutaneous vaccination with their husbands lymphocytes. The mean weight of the infants at birth was 2,975 ± 540 g, including one infant who was small for dates. Physical development parameters for the first year were all within normal range.

Clinical evaluation of focal excision of myometrial lesion for treatment of invasive hydatidiform mole.

To determine the clinical significance of conservative surgical therapy, namely focal excision of myometrial molar deposits, 22 patients with invasive hydatidiform mole (HMI who had received the therapy were analysed for their postoperative clinical course and reproductive performance. They were operated on because of their prolonged HCG regression curve and the presence of abnormal shadows in the uterine wall revealed by pelvic angiography, ultrasonography and computerized tomography after evacuation of intra‐uterine molar tissue. A definitive diagnosis of invasive HM was established histopathologically in all of the extirpated materials. Seven of the patients were given postsurgical chemotherapy because of prolongation of their HCG decrease after the operation. The following items were emphasized as necessary criteria when selecting patients for surgery consisting in complete resection of the myometrial lesion: (1) urinary HCG titers below 10,000 IU/day; (2) no evidence of pulmonary metastatic involvement; or (3) metastases in the lungs, controlled with chemotherapy prior to the operation. Their reproductive performance was almost the same as that of comparable patients who were treated by chemotherapy alone.

Influence of immunotherapy on the cellular immunity of unexplained recurrent aborters

Changes in lymphocyte subsets in whole blood were analyzed sequentially by flow cytometry with an automated leukocyte differential system in 15 patients with unexplained recurrent spontaneous abortions, each of whom underwent vaccination(s) with her husbands lymphocytes. Mitogen responses of peripheral blood lymphocytes (PBL) were also examined in these patients. The reactivity of PBL against mitogens revealed no significant change in each patient before and after vaccination(s) with her husbands lymphocytes. The CD4:8 ratio was observed to decrease significantly during 22 and 28 days after the first vaccination with a significant increase in the percentage of T suppressor-cytotoxic (CD8) cells. This change was also observed after the second vaccination. The percentages of other subsets did not change significantly after vaccination(s). In 11 patients out of 15, the pregnancy continued successfully and correlated with a predominance of Ts/c (CD8) over TH/I (CD4) cells in the first trimester. These changes in lymphocyte subsets may indicate the induction of immune enhancing mechanisms and it is suggested that continuation of the predominance of Ts/c cells induced by immunotherapy might be important for the successful maintenance of pregnancy.

Blocking Effects of Maternal Serum‐IgG and Placental Eluate‐IgG on Materno‐Fetal Mixed Lymphocyte Reaction and Their Individual Specificity*

ABSTRACT: The purpose of this study is to clarify the problem regarding individual specificity of maternal serum immunoglobulin G (IgG) and placental eluate IgG that block the unidirectional mixed lymphocyte reaction (MLR) in which maternal and cord blood lymphocytes (lys) were used as responder and stimulator, respectively. Both maternal serum IgG and placental eluate IgG were found to include blocking antibodies (BA) which showed similar blocking effect on MLR. But, in the case of serum IgG, individual specificity of BA on MLR was observed to be less great in MLR done using whole lys or T‐enriched lys of the cord blood as stimulator, whereas it became greatly significant when MLR was done using B‐enriched lys of the cord blood as stimulator. This was the case even in MLR done by using unrelated maternal lys as responder. On the other hand, in the case of placental eluate, it was found to be greater when compared to serum IgG even where MLR was done using not only B lys but also T lys as stimulator. It is thus strongly suggested that BA include individual specific antibodies against fetal B lys having HLA‐D/DR locus determinants in addition to individually nonspecific antibodies common to pregnancy which may be produced to a “yet unknown fetal antigen.”

Immunotherapy with paternal lymphocytes preceding in vitro fertilization-embryo transfer for patients with repeated failure of embryo transfer

Immunotherapy with paternal lymphocytes for unexplained recurrent aborters was applied, preceding IVF-ET, for infertile women with repeated failure of ET. In one patient showing close histocompatibility with the husband, the subsequent new IVF-ET was successful under positive MLR-blocking antibody induced by immunotherapy.

Clinical Analysis of Intracranial Metastases in Gestational Choriocarcinoma: A Series of 15 Cases

Summary: Between 1963 and 1980 we have had experience of 54 patients with gestational choriocarcinoma and 29 are alive, a survival rate of 53.7%. The disease had metastasized to the central nervous system in 15 of these 54 patients, an occurrence rate of cerebral metastases of 27.8%. 13 of these 15 patients have died and 2 are now in a state of remission. Autopsy was performed in 8 cases. The following points are emphasized: 1) cases with the pulmonary disease risk intracranial spread; 2) the serum/cerebrospinal fluid ratio of human chorionic gonadotropin concentrations and computerized tomography are recommended as useful investigations in the diagnosis of CNS disease; 3) decompression surgery in patients with increased intracranial pressure is indispensable; 4) radiotherapy is temporarily effective for brain deposits; 5) powerful systemic chemotherapy using multiple anticancer drugs is suggested to be helpful both in preventing and in treating CNS disease.