Charles R. King

Evaluation of human chorionic trophoblast cells and placental macrophages as stimulators of maternal lymphocyte proliferation in vitro

Dispersed cell suspensions of human chorion membrane and placentae were obtained by enzyme digestion and the cells examined for HLA expression and for the ability to stimulate immune cell proliferation in vitro. Chorion cells with the characteristics of trophoblast were HLA-A, B, C and Ia negative following tissue digestion whereas placental cells, primarily Fc gamma R positive macrophages, were HLA-A, B, C positive and were frequently Ia positive. When chorion and placental cell suspensions were used as stimulator cells in one-way mixed cell cultures (MCC) with maternal mononuclear leukocytes (MNL) as responder cells, chorion cells were not normally stimulatory (mean stimulation index (SI), 2.7) whereas placental cells usually were (mean SI, 11.5). No evidence for active suppression by chorion cells was obtained in a group of experiments designed to detect suppressive activity. The results support the concept of the trophoblast layer as an immunologically inert barrier between the mother and the fetus.

Determination of concentrations of adenosine and other purines in human term placenta by reversed-phase high-performance liquid chromatography with photodiode-array detection : evidence for pathways of purine metabolism in the placenta

A robust analytical method, using reversed-phase high-performance liquid chromatography with gradient elution and photodiode-array detection, was used to measure six purines and beta-NAD+ in acid-soluble extracts of samples taken from six different regions of human term placenta. Resolution of the analyte peaks in chromatographic profiles of the extracts, and the use of optimized integration, allowed simultaneous quantitation of all seven analytes from a single chromatogram. Peak purity was confirmed via on-line analysis of peak spectra, utilizing the purity parameter treatment of spectral data. Major placental purines were adenosine, inosine, hypoxanthine and adenine. Except for adenine, concentrations of the purines varied by two-fold or more between different regions of each placenta, but concentration ratios, i.e., adenosine/inosine and inosine/hypoxanthine, were similar. The findings indicate that the pathway of ATP breakdown to hypoxanthine in ischemic human term placenta is via adenosine, and that regional differences in placental concentrations of adenosine and its metabolites may result from regional differences in degree of ischemia.

Detection of class I MHC mRNA in subpopulations of first trimester cytotrophoblast cells by in situ hybridization

Class I MHC mRNA has been identified in both villous and extravillous cytotrophoblast cells in first trimester placentas by in situ hybridization. In this report, we expand those observations to additional morphologically and anatomically distinct subpopulations of trophoblast cells in early placentas using the same experimental approach. In the transition zone of first trimester placental villi, where cytotrophoblast cells are proliferating to form new villi or to migrate into adjacent tissue, both cytotrophoblast cells beneath the uninterrupted syncytium and the proliferating cytotrophoblast cells contained class I mRNA whereas a layer of cytotrophoblast cells proximal to the villus core did not contain class I mRNA. In the placental bed, migrating cytotrophoblast cells contained high levels of class I mRNA as determined by the intensity of staining. In contrast, multinucleated giant trophoblast cells contained little specific message. Alterations in levels of class I mRNA seem therefore to be associated with trophoblast proliferation, migration and differentiation.

Measurement of adenosine, inosine and hypoxanthine in human term placenta by reversed-phase high-performance liquid chromatography

An analytical method was developed for measuring adenosine, inosine and hypoxanthine in freshly delivered human term placentas. Representative freeze-clamped samples were taken from the sub-maternal surface of each placenta. Acid-soluble extracts of the samples were analyzed by reversed-phase high-performance liquid chromatography on columns packed with 10-micron porous octadecylsilica, using gradient elution with a linear increase in methanol concentration in ammonium phosphate buffer. Resolution of hypoxanthine from xanthine and adenosine from adenine, and quantitation of hypoxanthine and adenosine were achieved using 0.05 M ammonium dihydrogen phosphate, pH 6.5, as the low-strength eluent. Resolution of inosine from a prominent peak of beta-NAD was optimized using 0.02 M ammonium dihydrogen phosphate, pH 5.6, as low-strength eluent. Recovery of standards was greater than 90%. Mean contents (+/- S.D.) of the analytes in placentas from seven normal deliveries were, adenosine 30.6 +/- 11.5 nmol/g, inosine 68.0 +/- 25.8 nmol/g and hypoxanthine 217 +/- 127.5 nmol/g.

Ontogeny and distribution of cells expressing HLA-B locus-specific determinants in the placenta and extraplacental membranes

Both the trophectoderm and the inner cell mass of the blastocyst contribute to the cell populations found in the placenta and extraplacental membranes. Previous studies have shown differences between those two embryologically distinct populations of cells in their expression of class I HLA, and further differences among trophectoderm-derived trophoblast cell subpopulations. Binding patterns for antibodies to both monomorphic and allotypic determinants on class I heavy chains have been reported. In the present study, extraembryonic cells were evaluated by immunohistology for binding of the monoclonal antibody 4E, which identifies locus-specific determinants on HLA-B. Some inner cell mass-derived cells (mesenchymal cells) acquired high levels of HLA-B as gestation progressed and other continued to express low levels at late stages of gestation (amnion cells). In contrast, throughout gestation both villous and extravillous trophoblast cells failed to express detectable HLA-B. The binding patterns for 4E followed the patterns established by a monoclonal antibody to class I HLA heavy chains (61D2), and those reported for antisera to allotypic determinants. The findings support the suggestion that trophoblast cells forming the fetal component of the maternal-fetal interface exert highly effective regulation over the expression of class I HLA.

Diagnosis of congenital syphilis by immunofluorescence following fetal death in utero

The etiology of fetal death may be difficult to identify, particularly with the presence of marked fetal maceration and autolysis. Immunofluorescence with human antitreponemal antibody was used in this case to establish a diagnosis of congenital syphilis.

Abortion in Nineteenth Century America: A Conflict Between Women and Their Physicians

During the 1800s, male physicians took over womens established sources of medical care and knowledge causing friction between women and their health matters and the rising medical profession. This conflict still exists today. Physicians provided little information on how to prevent pregnancy since to do so would interfere with their natural biologic function of motherhood. Reproduction no longer controlled women who used contraceptives, had an abortion, and those who supported such decisions. Since women had to actively terminate a pregnancy themselves or seek an abortionist, abortion posed a threat to the traditional male dominance of reproductive decisions. Even though no accurate data on the frequently of abortion existed, women often practice abortion in the absence of other means of fertility control. They even helped their fellow sisters by telling them what they can use to induce abortion. These women were often church-going women and well respected in their communities. During most of the century, the clergy did not condemn abortion. There were plenty of agents and materials available to induce abortion which were sometimes publicized as methods to regulate menstruation. Abortifacients included emmenagogues, purgatives, oxytocics, and herbal or botanical products. Yet many proprietary potions did not actually terminate pregnancies. Women often resorted to using sharp instruments, wax candles, penholders with attached wires, glass rods, hair curling tongues, sticks, spoons, knives, and catheters. Physicians during the 19th century did not have the diagnostics means to determine pregnancy so often women misled them so the physicians would perform medical interventions. In the mid-1800s, allopathic physicians strengthened their professional position by regulating physicians and forming the American Medical Association. This resulted in condemnation of abortionists and abortions. They blamed complications of abortion on womens reproductive decisions. The new political climate concerning abortion resulted in regulation of women and their sexuality.