Identification of selectively solubilised syncytiotrophoblast plasma membrane proteins as potential antigenic targets during normal human pregnancy.

Syncytiotrophoblast plasma membranes prepared from term placentae were selectively solubilised in non-ionic detergents. The solubilised proteins and the insoluble residue were tested in an ELISA assay for their ability to function as antigenic targets for anti-trophoblast antibodies present in normal first trimester pregnancy sera. The soluble proteins were fractionated by gel filtration and four major antigen forms were identified. The antigens were reactive with affinity purified anti-trophoblast antibody isolated from maternal sera and hence were termed maternally-recognised trophoblast antigens (MRTA); these were designated MRTA-I (Mr = 400,000 D), MRTA-II (Mr = 142,000), MRTA-III (Mr = 50,000) and MRTA-IV (Mr = 13,000). The relationship between MRTA-I, II, III and IV and antigens identified in maternal sera in the form of immune complexes is discussed.

Anti-trophoblast antibody responses during normal human pregnancy.

During the course of a normal uncomplicated human pregnancy the mother generates an antibody response directed against determinants present on the plasma membrane of the outer fetal layer of the term placenta, the syncytiotrophoblast. The response, measured by an ELISA that utilises syncytiotrophoblast plasma membrane as the antigenic target, is predominantly IgG in nature, but with a minor contribution from IgM molecules. Maximum responses were observed during the first trimester and the levels gradually declined as the pregnancy progressed. On a population basis, this antibody response profile was mainly restricted to first and second pregnancies, although anti-trophoblast antibody responses could be detected in multiparous women but with a greatly reduced incidence compared with primipara. Mechanisms to account for these observations are discussed. Throughout, the anti-trophoblast antibody levels detected in pregnancy sera were compared with the background levels which were observed in sera obtained from males and nulliparous non-pregnant females.

Pregnancy-associated nonspecific immunosuppression: mechanism for the activation of the immunosuppressive factors.

The nonspecific immunosuppressive effect observed in pregnancy sera and mediated by two factors, immunosuppressive factors (ISF) I and II, was regulated by a third molecule termed the pregnancy-depleted immunoregulatory factor (pdIRF). Natural depletion of serum pdIRF levels during pregnancy resulted in the activation of the ISF-I and ISF-II molecules, which prior to conception existed in the serum in inactive forms. The inactive ISF-I and ISF-II molecules in male sera and in nulliparous nonpregnant female sera can be activated following the artificial selective depletion of pdIRF by absorption onto Sephacryl S-300. It is proposed that inactive complexes consisting of the ISF-I and ISF-II molecules and pdIRF and possibly involving Ca2+ and Mg2+ ions are a feature of normal sera. The pdIRF molecule has an apparent Mr of 100,000-125,000 daltons and consists of a single major polypeptide of 63,000 daltons. Equal concentrations of pdIRF are present in male and female normal sera.

Pregnancy-associated nonspecific immunosuppression: kinetics of the generation and identification of the active factors.

The effect of gestational age and maternal parity on the development of nonspecific immunosuppressive activity in the sera of pregnant women, which inhibited the in vitro transformation of unrelated lymphocytes by phytohemagglutinin, was examined. Quantitative demonstration of this activity was dependent, in part, on the source of the lymphocytes and on the serum concentration in culture. The immunosuppressive activity became evident as the pregnancy progressed, and in late-pregnancy sera it was mediated by two factors, immunosuppressive factors (ISF) I and II with apparent Mr of 2 X 10(6) and 150,000 daltons. By analysis and comparison of different types of sera fractionated by gel filtration on Sephacryl S-300, it was evident that ISF-I and ISF-II were also present in male and nulliparous nonpregnant female sera, but in inactive forms. Hence the immunosuppressive factors did not appear to be "produced" in pregnancy, but the observed activity was a reflection of the "activation" of preexisting molecules in the serum. An accompanying report (Am J Reprod Immunol Microbiol. 1985; 9:84-90) describes the regulation of the activation event.

Localization of immunoglobulin-containing cells in human endometrium in the first trimester of pregnancy and throughout the menstrual cycle.

The distribution of immunoglobulins in normal human endometrium throughout the menstrual cycle and in early pregnancy has been studied with an immunoperoxidase technique. In first-trimester decidua, IgG was detected within many cells of differing morphology and size. Large IgG-containing cells were often binucleate and were believed to be decidual cells. Examination of serial sections showed no kappa or lambda light-chain restriction, suggesting absorption of the immunoglobulin content. Medium-sized, irregular, IgG-containing cells were macrophages. An additional substantial population of small hyperchromatic IgG-containing cells were prominent around arterioles and adjacent to endometrial glands. From examination of adjacent sections stained with phloxine tartrazine, it was concluded that these represented endometrial granulocytes. Labelling for light chains again suggested absorption of the immunoglobulin content. In contrast, in non-pregnant endometrium immunoglobulin-containing stromal cells were uncommon, although IgG and IgA were detected in gland epithelium and secretions and in the stromal interstitium particularly in the secretory phase. These results support the notion that human endometrium lacks a classical secretory immune system and highlight the requirement for correlation between studies of cell surface markers, morphology and cell surface receptors.

The partial characterisation of maternal anti-trophoblast antibody responses generated during normal human pregnancy.

The anti-trophoblast antibody response generated during a normal human pregnancy and detected by a recently developed enzyme-linked immunosorbent assay, was partially characterised in terms of maternal influences, nature of the antibodies and nature of the antigenic determinants present on the syncytiotrophoblast plasma membrane. The level and incidence of the response was significantly affected by maternal parity, while the maternal ABO, but not Rhesus, blood group antigens exerted a minor influence. The antibody response was predominantly mediated by IgG molecules of the IgG1,2 and 4 subclasses. The IgG molecules existed in the maternal sera either in the form of 'free' molecules or were involved in immune complexes. The antibodies interacted with determinants that were present on all the placental membranes tested and hence are possibly organ specific. The antigenic specificities were absent from erythrocytes and peripheral blood lymphocytes.

Transcription of individual genes in eukaryotic cells occurs randomly and infrequently.

Experimental evidence is presented indicating that the expression of a lacZ reporter gene driven by the HIV-1 long terminal repeat in a series of stably transfected, cloned macrophage cell lines occurs in a very small proportion of cells. The proportion of cells expressing lacZ, rather than the level of expression in each cell, is regulated by external stimuli such as LPS and phorbol ester. Based upon these and published data we propose that transcription in eukaryotic cells occurs in short pulses interspersed by long periods of inactivity of indeterminate duration. Transcriptional regulation is envisaged as involving changes in the probability rather than the rate of transcription. A probabilistic model of transcription may explain many biological phenomena, such as stem cell division and clonogenic activity, heterogeneous gene expression among clonal cell populations, retroviral latency and cell cycle progression, which appear to involve stochastic decisions.

Differentiation of the mononuclear phagocyte system during mouse embryogenesis: the role of transcription factor PU.1.

During mouse embryogenesis, macrophage-like cells arise first in the yolk sac and are produced subsequently in the liver. The onset of liver hematopoiesis is associated with the transition from primitive to definitive erythrocyte production. This report addresses the hypothesis that a similar transition in phenotype occurs in myelopoiesis. We have used whole mount in situ hybridization to detect macrophage-specific genes expressed during mouse development. The mouse c-fms mRNA, encoding the receptor for macrophage colony-stimulating factor (CSF-1), was expressed on phagocytic cells in the yolk sac and throughout the embryo before the onset of liver hematopoiesis. Similar cells were detected using the mannose receptor, the complement receptor (CR3), or the Microphthalmia transcription factor (MITF) as mRNA markers. By contrast, other markers including the F4/80 antigen, the macrophage scavenger receptor, the S-100 proteins, S100A8 and S100A9, and the secretory product lysozyme appeared later in development and appeared restricted to only a subset of c-fms-positive cells. Two-color immunolabeling on disaggregated cells confirmed that CR3 and c-fms proteins are expressed on the same cells. Among the genes appearing later in development was the macrophage-restricted transcription factor, PU.1, which has been shown to be required for normal adult myelopoiesis. Mice with null mutations in PU.1 had normal numbers of c-fms-positive phagocytes at 11.5dpc. PU.1(-/-) embryonic stem cells were able to give rise to macrophage-like cells after cultivation in vitro. The results support previous evidence that yolk sac-derived fetal phagocytes are functionally distinct from those arising in the liver and develop via a different pathway.