Maternal alloimmune reactions towards the murine conceptus and graft-versus-host reaction (GVHR). I. Priming for anti-paternal GVHR by gestation

In the H-2 compatible (but minor loci-incompatible) BALB/c-DBA/2 strain combination (both H-2d), intravenous injection of 1.3 X 10(7) BALB/c spleen cells from virgin females into DBA/2 newborn mice less than 18 h old does not result in a significant lethal graft-versus-host reaction (GVHR). A strong GVHR (79% lethal) is induced if the BALB/c donors have been preimmunized to DBA/2. Spleen cells from BALB/c mice pregnant by DBA/2 males are also able to induce a significant, but weaker, GVHR (16% lethal) indicating a cellular priming to paternal antigens by gestation. A significant difference exists between anti-DBA/2 GVH reactivity of spleen cells from primiparous (22% lethal) and multiparous (9% lethal) allopregnant BALB/c mice, indicating that the allogeneic boosters of successive allogestations act more on the target-protective side of immunity than on the target-aggressive one. Sera from allopregnant mice (BALB/c X DBA/2) inhibit the GVHR induced by their own cells, while sera from isopregnant ones (BALB/c X BALB/c) have no effect. Thymectomy performed at 6-wk of age, six weeks before gestation did not significantly modify the maternal reactivity. A similar priming by allogestation in the same strain combination was found for local GVHR (induced in adult F1 hybrids) resulting in higher (+132%, P less than 0.005) stimulation indices and seen to be specific for the paternal strain, the indices induced by the same cells being lower (-35%, P less than 0.05) compared to that induced by cells from virgin BALB/c, when injected into irrelevant F1 hybrids (BALB/c X CBA).     

Deviation of humoral and cellular alloimmune reactions by placental extracts

Modifications of the alloimmune response at both the humoral and the cellular levels by placental extracts (PE) syngeneic to the recipient were studied in the mouse using two different H-2 strain combinations. CBA (H-2k) or C57BL/Ks (H-2d), immunized with A/J (H-2a) spleen cells. The tests included in vivo tumor allograft evolution (accelerated rejection or enhancement reactions), and in vitro analysis of the involved immune agents, both cellular and humoral, using mixed lymphocyte reactions (MLR) and biological activity studies of serum samples. Animals from the recipient strains exhibited a delayed rejection of A/J tumor Sa 1 allografts if preimmunization was carried out with 10(6) A/J spleen cells combined with PE syngeneic to the recipients, as compared to controls immunized with A/J cells only or supplemented with isogeneic liver extracts (LE). The serological analysis revealed that PE treatment did not modify the overall hemagglutinating antibody production but resulted simultaneously in both a decreased production of cytotoxic complement fixing antibodies and an increase of specific anaphylactic mast cell degranulating antibodies, as compared to controls. The sera from PE-treated donors also demonstrated enhancing activity following passive transfer to isogeneic recipients. MLR regulatory activity was exhibited by spleen cells from PE- and immunogen-treated mice although the same or stronger activity was obtained from mice immunized without the addition of PE. However, in vivo transfer of these cells to syngeneic recipients showed that PE treatment erased the accelerated rejection caused by allogeneic immunization in the absence of PE and could even cause some degree of allografted tumor enhancement. The cells responsible for this inhibitory effect were mainly IJ+ lymphocytes, since their elimination with a relevant anti-IJ serum and complement restored a secondary type rejection pattern. These results show that PE present during the onset of immunization can promote the activation of regulatory agents such as enhancing antibodies and suppressor cells favoring allograft survival.     

Inhibition of ovarian, placental, and adrenal steroidogenesis in the rhesus monkey by trilostane.

Trilostane inhibits adrenal, ovarian, and placental steroidogenesis when administered orally to rhesus monkeys. By inhibiting 3 beta-hydroxysteroid dehydrogenase activity, it causes an increase in circulating levels of pregnenolone. Trilostane reverses the stimulation of luteal progesterone production and the delay in onset on menstruation induced by human chorionic gonadotropin. In pregnant monkeys it reduces circulating progesterone levels and is an effective interceptive agent if given for 5 days beginning on day 16, 25, or 50 of gestation. Concurrent administration of progesterone prevents this interceptive effect. Trilostane reduces plasma cortisol levels at doses lower than those necessary to terminate pregnancy.     

Inhibition by tiazofurin of inosine 5'-phosphate dehydrogenase (IMP DH) activity in extracts of ovarian carcinomas.

Cancer cells have an increased ability to synthesize GTP (guanosine triphosphate) because of increased activity of IMP DH (inosine 5'-phosphate dehydrogenase, EC 1.1.1.205). Because IMP DH activity is rate limiting for de novo biosynthesis of GTP, this enzyme was suggested as a sensitive target for chemotherapy. Tiazofurin (2-beta-D-ribofuranosylthiazole-4-carboxamide) is converted in the cells into the active metabolite, TAD, (thiazole-4-carboxamide adenine dinucleotide) which potently inhibits IMP DH activity. By adding TAD to tissue extracts one can determine the extent of inhibition of IMP DH. We applied the IMP DH assay method to extracts of normal ovaries (N = 11) and epithelial ovarian carcinomas (N = 10). The IMP DH activity (mean +/- SE) in ovarian carcinoma was 21.1 +/- 5.8 which was markedly higher than that observed in normal ovaries (2.9 +/- 0.7 nmol/hr/mg protein) (P < 0.05%). The inhibition by TAD of IMP DH activity in ovarian carcinomas (N = 4) was 81%. The results indicate that IMP DH activity is elevated sevenfold in ovarian carcinomas as compared to normal ovary and can be inhibited by exposure to tiazofurin (TAD). Similar high IMP DH activity and inhibition of the activity by TAD was observed in patients with chronic granulocytic leukemia in blast crisis among whom 70 to 80% remissions were reported. Since there is increased IMP DH activity in human ovarian carcinomas and in OVCAR-5 cells and tiazofurin and TAD inhibit IMP DH activity of these cells and the proliferation of human ovarian carcinoma xenografts in the mouse, tiazofurin may merit serious consideration for a Phase II trial for patients with recurrent/refractory epithelial ovarian carcinoma.     

Maternal serum concentration of placental growth factor (PIGF) and endothelial growth factor (VEGF) in pregnancies complicated by preeclampsia

Preeclampsia is one of the most frequent and dangerous complications of a pregnancy. In preeclamptic pregnancies the spiral arteries are not modified properly. Disturbed blood flow finally leads to hypoxia which is responsible for the dysfunction of the endothelium. Endothelial growth factor (VEGF) and placental growth factor (PIGF) play an important role in the angiogenesis and thus may participate in the pathomechanism of preeclampsia. AIM: The aim of our study was to estimate VEGF and PIGF level in serum of patients with preeclampsia. MATERIALS AND METHODS: The study comprised 25 gravidas with preeclampsia and a control group of 18 healthy gravidas. In 25 preeclamptic women the angiogenic factors levels were measured in the III trimester and in 7 of them in the II trimester. In the control group these parameters were assessed in both periods. Both factors were measured by commercial available ELISA KIT. RESULTS: PIGF concentrations were significantly (p < 0.0001) lower in sera of patients with preeclampsia in the II and III trimester in comparison to the controls: 17.4 vs. 290.3 pg/ml and 99.1 vs. 347.8 pg/ml, respectively. In most cases serum VEGF levels were undetectable. CONCLUSIONS: (1) PIGF is involved in the pathomechanism of preeclampsia and its maternal serum concentration decreases significantly in the course of the disease. (2) The sensitivity of the commercially available ELISA assay is too low to assess the serum VEGF concentration in women with preeclampsia.     

Chorangiomas (hemangiomas of the placenta). II. Pathologic-anatomic comparison of neonatal complications

Fetal and neonatal complications in chorangiomas of different size, in multiple chorangiomas and in cases accompanied with hydramnios were reviewed in 110 cases from literature inclusively 7 own observations. In accordance with other examiners we found an increasing perinatal mortality, especially of the antenatal mortality rate corresponding with an increasing tumor size. Chorangiomas with hydramnios had a mortality of 80 per cent. In these cases only a small part was caused by fetal immaturity or severe malformations. Comparing mean placental weight and mean neonatal weight small for date babies had a relative and absolute augmentation of placental tissue in relation to a control group. The conclusion from these findings and the fact that in own cases heavy disorders of villous differentiation could be observed is, that the fetal fate may be fundamentally influenced not only by the hemodynamic disturbance caused by the tumor, but also by the degree of placental differentiation. Large hemangiomas are ought to detect during pregnancy by ultrasound. In singular cases it may be to prevent fetal hypoxia by termination of pregnancy.     

Humoral immune responses in murine pregnancy. V. Relationship to the differential immunogenicity of placental and fetal tissues

The nature of the humoral immune response induced in virgin female mice by injections of F1 placental and fetal tissues has been examined and compared to that induced by immunization with F1 adult spleen cells and by multiple allogeneic pregnancy. In a 'responder' strain mouse, as defined by the ability of multiple allogeneic pregnancy to elicit an anti-paternal humoral immune response, both F1 placental and fetal tissues induced the formation of alloantibodies primarily of the IgG1 sub-class, similar to those induced by allogeneic pregnancy, but different from those elicited by adult spleen cells. However, only the placental tissues induced alloantibodies possessing all the characteristics of those appearing in multiparous allogeneic pregnancy. In contrast, the alloantibodies induced by the injected fetal tissue possessed complement-dependent cytotoxic activity, indicating that the inability of pregnancy-induced alloantibodies to mediate cytotoxicity may not be related to their restriction to the IgG1 sub-class. In a 'non-responder' mouse strain, where multiple allogeneic pregnancy does not lead to a maternal alloantibody response, F1 placental tissues, in contrast to fetal and adult tissues, failed to induce a humoral immune response. Injection of F1 placental tissue therefore elicits responses that mimic both the properties and the strain-dependent distribution of the alloantibodies identified in normal murine pregnancy. This implies that the immunogenic stimulus in pregnancy emanates from the placental rather than the fetal compartment of the allogeneic conceptus.     

Immunosuppression by human placenta lactogen (HPL) and the pregnancy-specific beta 1-glycoprotein (SP-1). Inhibition of mitogen-induced lymphocyte transformation

The in vitro effect of human placental lactogen (HPL) and the gravidity-specific beta1-Glycoprotein (SP-1) on the lymphocyte transformation was investigated in 20 healthy women. It was shown that simultaneous incubation of the lymphocytes with HPL or SP-1 together with PHA has no influence on the incorporation rate of H3-Thymidine. When the fetal calf serum (FCS) in the lymphocyte cultures was substituted by a 3% human albumin-solution, there was a reduction of the incorporation rate of the radioactive nucleic acid precursors to 20%, as compared to the controls with FCS. After addition of HPL and SP-1 to the cultures there was a further dosis dependent decrease of the incorporation rate. When lymphocytes were preincubated with the proteins for 24 h, there was a marked reduction of the blastogenesis after additon of PHA, HPL and SP-1 under these conditions had a immunosuppressive effect on the lymphocyte transformation.     

Inhibition by plant herb extracts of steroid bindings in uterus, liver and serum of the rabbit

To assess the action of herb extracts, glycyrrhizin and paeoniflorin, upon steroids, their binding to several classes of intracellular and serum steroid-binding proteins was studied in the rabbit. Glycyrrhizin and paeoniflorin exhibited similar binding behaviors. They bound minimally to estrogen and androgen receptors, but not to the progesterone receptor in uterine cytosol, and exhibited a moderate binding activity to glucocorticoid receptors in liver cytosol. They also exhibited weak binding activity to both cortico-steroid-binding globulin and sex-hormone-binding globulin.     

Proteins of human placental microvilli: II. Identification and topology of the plasma membrane proteins

We have investigated the location of proteins in the transverse plane of the plasma membrane of microvilli isolated from the syncytiotrophoblast layer of the human term placenta. Microvillous proteins were labelled with 125I under reaction conditions where those proteins exposed on the maternal-facing surface of the microvilli were most heavily labelled. The proteins were then solubilized and subjected to one- and two-dimensional electrophoresis followed by protein staining and autoradiography. More than 65 proteins, differing in molecular weight or isoelectric point or both, were identified, and these were classified into three groups: weakly, moderately heavily, and heavily labelled. The microvilli were in the form of intact vesicles that were correctly orientated ('right-side out'). Thus the extent of labelling of each protein could be used as an indication of the extent of its exposure on the maternal-facing surface of the microvilli. Human serum albumin was present on the surface of the isolated, washed microvilli, but was probably a contaminant originating from maternal blood.     

Maternal insertion of 18q11.2-q12.2 in 18p11.3 of the same chromosome analysed by microdissection and multicolour banding (MCB).

OBJECTIVES: Different aberrations in one chromosome 18 were prenatally detected during each of three different pregnancies of a healthy woman. Routine cytogenetic analysis revealed a morphologically altered maternal chromosome 18 as well. The purpose of the current study was to characterize these cytogenetic changes in detail and thus to clarify the reason for the recurrent appearance of morphologically altered chromosomes 18 in this family. METHODS: As GTG banding did not allow resolution of the kind of aberrations present in these four cases, the following molecular cytogenetic approaches were used: microdissection combined with reverse painting and multicolour banding (MCB) analysis using a chromosome 18 specific probe set. RESULTS: Molecular cytogenetic approaches revealed that fetus 1 had a derivative chromosome del(18)(q11.2q12.2), fetus 2 and the mother had the identical derivative chromosomes ins(18)(pterp11.32::q12.2q11.2::p11.32q11.2::q12.3qter) and fetus 3 had a dup(11.2q12.2). CONCLUSION: Partial monosomy in fetus 1 and partial trisomy in fetus 3 can be explained by crossing over events during maternal meiosis.     

Attitude of Saudi families affected with hemoglobinopathies towards prenatal screening and abortion and the influence of religious ruling (Fatwa).

Hemoglobinopathies are common inherited disorders in Saudi Arabia. Prenatal diagnosis for such diseases is specific and sensitive but not yet implemented in Saudi Arabia. Saudis are Muslims with a very high rate of consanguinity and inherited genetic disorders. To examine the attitude of Saudi families affected with hemoglobinopathies towards prenatal diagnosis and abortion, and to evaluate the effect of education on religious ruling on such attitudes, 32 families were interviewed using a pre-structured questionnaire. The majority accepted prenatal diagnosis (81.3%). The attitude towards abortion was greatly affected by religious values. Education about religious ruling significantly affected parents' attitude towards accepting abortion and prenatal diagnosis. No other factors were found to influence the outcome. Although the majority of families received some kind of formal genetic counseling [23/32 (71.9%)], none of them was informed about the possibility of prenatal or preimplantation diagnosis prior to the interview. Therefore for prevention of genetic disorders, the emphasis in countries with a vast majority of Muslims such as Saudi Arabia has probably to be placed on public awareness about genetic risks, the risk of consanguinity, availability of services, and so on, while at the same time taking into consideration the religious beliefs and education of the target population