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PubMed1986

Presence of an endothelial antigen on the syncytiotrophoblast of human chorionic villi: detection by a monoclonal antibody.

We describe a monoclonal antibody, B721. This antibody reacts with an antigen present on vascular endothelium and on the syncytiotrophoblast of term chorionic villi. The antigen is absent from the trophoblast of the chorion, from the amniotic epithelium, and from normal peripheral blood or lymph node lymphocytes. We discuss the possible functional roles of the antigen. We propose that the syncytiotrophoblast, by expressing endothelial antigens, mimics endothelium and may perform endothelial functions.

Adult↗
PubMed1995

Maternal cells in chorionic villi from placentae of normal and abnormal human pregnancies.

PROBLEM: We asked if activated macrophages and CD4 positive T lymphocytes in placental chorionic villi with villitis were of maternal or fetal origin. METHOD: We employed a double antibody immunocytochemical technique on placental sections from three normal and four abnormal pregnancies with small-for-gestational-age infants. All studied placentae were mismatched for the maternal-fetal HLA-DRw 52 antigen. Areas of immunopathology were identified by using a monoclonal antibody to a monomorphic determination on HLA-DR, and the origin of immunological cells in areas of immunopathology was identified by using a monoclonal antibody to a polymorphic determinant on HLA-DRw 52. RESULTS: We used a double antibody technique that employed monoclonal antibodies to HLA-DR and HLA-DRw 52 antigens and placentae that were mismatched for the maternal-fetal HLA-DRw 52 antigen. We found that the vast majority of immunological cells within villi with inflammation were of maternal origin. Quantitative studies showed that between 75 and 100% of the cells in normal as well as in abnormal pregnancies were of maternal origin, and that abnormal pregnancies had a significantly higher percentage of villi with maternal cellular infiltrates. CONCLUSION: Our data show unequivocally that cells in areas of placental immunopathology are predominantly of maternal origin, and that abnormal pregnancies are associated with significantly more villi containing immunological cells of maternal origin.

Adult↗
PubMedDate not supplied

Exclusion of citrullinaemia in the first trimester of pregnancy by direct assay of argininosuccinate synthetase in chorionic villi.

Citrullinaemia was presumed to be excluded in a fetus at risk by the direct assay of argininosuccinate synthetase in chorionic villi. The diagnosis was confirmed after amniocentesis by normal argininosuccinate synthetase activity in the cultured amniotic fluid cells and by a normal citrulline concentration in the amniotic fluid. The prediction of a normal fetus was confirmed at term by the birth of a non-citrullinaemic boy.

Amino Acid Metabolism, Inborn Errors↗
PubMed1986

DNA-diagnosis of sickle cell anemia from chorionic villi: possible influence of maternal cell contamination.

The admixture of maternal tissue is a possible hazard of prenatal diagnosis from chorionic villi. Therefore the tolerable degree of maternal DNA contamination in prenatal diagnosis of sickle cell anemia was investigated by mixing various amounts of DNA from HbS carriers with DNA from normal individuals and sickle cell anemia patients. The results indicate that lower rate of admixed maternal DNA does not prevent an exact direct DNA-diagnosis of sickle cell anemia.

Anemia, Sickle Cell↗
PubMed1977

Immunological studies of human placentae: the distribution and character of immunoglobulins in chorionic villi.

All four human IgG subclasses, and both kappa and lambda light chains, were detected by immunofluorescence in similar distributions in chorionic villi of human placentae. IgG1 and IgG3 were the predominant subclasses. No evidence was obtained for local enzymatic digestion of IgG during placental transfer. Most of the IgG on the trophoblastic basement membrane (TBM) was loosely bound and could be removed by prolonged washing, although some appeared to be more tightly bound to small segments of the TBM. IgM, but not IgA, was present in small amounts in placental villous structures. Immunoglobulin was never observed within the syncytiotrophoblast. Antisera to IgG genetic (Gm) markers were used to locate IgG thought to be of foetal or maternal origin. The presence of paternal Gm markers not carried by the mother was taken as evidence for foetal IgG. Foetal (paternal) Gm markers were observed in placentae, although maternal IgG was the major immunoglobulin present in placental villi. Both maternal and foetal IgG were demonstrated in fibrinoid deposits, vessel walls and the cytoplasm of some stromal cells. Only foetal IgG was definitively observed in the immunoglobulin that is tightly bound to the TBM.

Basement Membrane↗
PubMed1996

Ketoacids attenuate glucose uptake in human trophoblasts isolated from first-trimester chorionic villi.

OBJECTIVE: The objective of this study was to evaluate the hypothesis that ketoacids (acetoacetic acid and beta-hydroxybutyrate) diminish glucose transport in trophoblasts cultured from first-trimester chorionic villi. STUDY DESIGN: First-trimester trophoblasts were obtained by transabdominal chorionic villus sampling for subsequent cytogenetic analysis. The cells were established as a continuous line exhibiting trophoblast characteristics. Trophoblasts were cultured in Ham's F12/Dulbecco's modified Eagle's medium (1:1) supplemented with 15% fetal bovine serum. Experiments were initiated by a 24-hour preincubation in serum-free Ham's F12/Dulbecco's modified Eagle's medium followed by incubation with ketoacids (acetoacetic acid and beta-hydroxybutyrate, 0 to 10 mmol/L) in the presence or absence of insulin-like growth factor-I (100 ng/ml). The cells were challenged with 2-deoxy-[1,2-3H]D-glucose (0.1 mmol/L) for 5 minutes and then cell-associated radioactivity was measured. Total ribonucleic acid was extracted from cells incubated with ketoacids in the presence or absence of insulin-like growth factor-I, and Northern blots were probed with a phosphorus 32-labeled complementary deoxyribonucleic acid fragment encoding the rat GLUT 1. RESULTS: Ketoacids caused a dose-dependent inhibition of glucose transport. At 5 mmol/L acetoacetic acid there was a > 50% reduction in the rate of glucose transport in both control and insulin-like growth factor-I-treated cells. The diminution in glucose uptake by trophoblasts was not due to cellular toxicity of the ketoacids because there was no significant difference in trypan blue exclusion or lactate dehydrogenase release between control and ketoacid-treated cells. Northern analysis revealed that the steady-state expression of GLUT1 messenger ribonucleic acid was diminished in ketone-treated cells, but this effect was overcome by coincubation of cultures with insulin or insulin-like growth factor-I. CONCLUSIONS: These results indicate that ketoacids can suppress the uptake of glucose into first-trimester human trophoblasts. Because ketoacidosis in pregnant women with diabetes mellitus is a frequent clinical consequence of poor metabolic control, it is possible that elevated levels of acetoacetic acid and beta-hydroxybutyrate may impair the transport of glucose across the placental trophoblast and into the fetal circulation.

Acetoacetates↗
PubMed1981

Binding of antigen-antibody complexes to mononuclear phagocytes in chorionic villi of human placentae.

Both endogenous placental IgG and exogenous antigen-antibody complexes are bound to stromal cells within the chorionic villi of human placentae. Both types of IgG are removed readily from placentae by washing at pH 5.0, while antibodies to placental antigens, when reacted with the villi, are not removed except under very low pH conditions (pH 2.5). Furthermore, when IgG is removed from term placentae and characterized by filtration chromatography, most of the IgG is seen to occur either as IgG fragments or as complexes of greater than 200000 relative molecular mass. This suggests that placental IgG exists as Fc gamma receptor-associated antigen-antibody complexes.

Adsorption↗
PubMed1987

Use of 4-methylumbelliferyl-6-sulpho-2-acetamido-2-deoxy-beta- D-glucopyranoside for prenatal diagnosis of Tay-Sachs disease using chorionic villi.

Prenatal diagnosis of Tay-Sachs disease was performed using the sulphated substrate 4-methylumbelliferyl-6-sulpho-2-acetamido-2-deoxy-beta-D-glucopyranoside to detect hexosaminidase A in chorionic villi. The presence or absence of hexosaminidase A in villi was detected by both a quantitative enzyme assay, and by a rapid, novel procedure which permitted visual discrimination between normal and affected villi.

Chorionic Villi↗
PubMed1988

Partial deletion of 4p in fetal cells not present in chorionic villi.

A case of a prenatal diagnosis at the second trimester is presented showing a normal karyotype in 12 metaphases from chorionic villi. In all cultured amniotic cells, however, and also in all fetal fibroblasts analyzed after abortion a structural anomaly (46,XY;del 4(pter----p15.2) was detected. Prenatal diagnosis was performed because of intrauterine growth retardation, cleft lip and esophagus atresia by ultrasound. The fetal stigmata are compatible with the Wolf Hirschhorn syndrome. We conclude that amniocentesis may be indicated notwithstanding a normal CV-diagnosis in those rare pregnancies with a characteristically abnormal ultrasound.

Abnormalities, Multiple↗
PubMed1993

Discrepancy in mosaic findings between chorionic villi and amniocytes: a diagnostic dilemma involving 45,X, 46,XY, and 47,XYY cell lines.

Discrepancies between cytogenetic findings in chorionic villi (CV) and fetal tissue have been reported. Several embryogenic models have been proposed to explain such discrepancies. We describe a case in which analysis of the direct preparation showed 24% 45,X and 76% 46,XY, with 43% 45,X and 57% 46,XY cells in cultured villi. Amniocentesis results disclosed 97% 46,XY and 3% 47,XYY. No 45,X cells were found in cultured amniocytes. These findings suggest that nondisjunction occurred early in postzygotic cleavage resulting in 3 cell lines. It is postulated that through selection, the less viable 45,X cells died out among those destined to become fetus proper but persisted among the cytotrophoblast and extraembryonic mesoderm cells. While there is probably selection against all aneuploid cell lines, 47,XYY cells are more likely to survive in the fetus. An explanation for the lack of 47,XYY cells in the CV might be simply that the tissue sampled was not representative of the cytogenetic make up of the entire placenta.

Adult↗
PubMed2000

Modulation of indoleamine 2,3-dioxygenase by interferon-gamma in human placental chorionic villi.

The effect of interferon-gamma on indoleamine 2,3-dioxygenase, a tryptophan catabolizing enzyme, was studied in cultured human placental chorionic villi. The activity of indoleamine 2,3-dioxygenase was markedly stimulated by interferon-gamma in a time- and concentration-dependent manner. Interferon-alpha and interferon-beta also showed a slight stimulatory effect on indoleamine 2,3-dioxygenase activity. The level of indoleamine 2,3-dioxygenase mRNA expression (determined by reverse transcription-polymerase chain reaction) was also enhanced by interferon-gamma. Interleukin-4 showed a dose-dependent inhibitory effect on interferon-gamma-induced stimulation of indoleamine 2,3-dioxygenase activity and mRNA expression.

Chorionic Villi↗
PubMed1992

Apparent non-mosaic trisomy 16 in chorionic villi: diagnostic dilemma or clinically significant finding?

A case is presented in which apparent non-mosaic trisomy 16 was found in chorionic villi (direct and culture) obtained from a patient undergoing first-trimester prenatal diagnosis. The fetal karyotype subsequently was determined to be 46,XX by follow-up amniocentesis. Serial ultrasonographic examinations revealed placental sonolucencies and intrauterine growth retardation. At 37 weeks, a small-for-gestational-age female was delivered by Caesarean section for fetal distress. Postnatal cytogenetic studies revealed a normal female karyotype in cord blood and mosaic trisomy 16 in placental tissues. These findings suggest that in cases where aneuploidy is confined to placental tissues, it may have biological significance, as evidenced by the apparent placental dysfunction and poor fetal growth in this case.

Adult↗
PubMed1990

Cell proliferation in chorionic villi at different gestational ages, as analyzed by premature chromosome condensation.

The technique of premature chromosome condensation (PCC) was adapted to human first-trimester chorionic villi cells to analyze the cell-cycle kinetics of interphase chromatin. Uncultured cells of the cytotrophoblast (CT) and the mesenchymal core (MC) were obtained by a two-step digestion. PCC was induced by fusion of the chorionic interphase cells with mitotic Chinese hamster ovary or HeLa cells. Cells showing PCC in G1 (classes 1-6), S, and G2 were found. To analyze further the proliferation stages of chorionic G1 interphases, the proliferation potential index (PPI) of 34 placentae recovered between the 8th and 12th week of gestation was determined. The mean PPI found in the CT and MC cells ranged from 18% to 73%, values similar to those described for intensely proliferating tissues. The highest mean PPI value (73%) was observed in CT cells from placentae recovered at the 9th week of gestation, indicating a high specific proliferative activity of CT cells at this developmental stage.

Animals↗
PubMed1984

DNA and enzyme studies on chorionic villi for use in antenatal diagnosis.

A study has been undertaken to determine the efficiency of current methods in providing an adequate amount of chorionic villus DNA for antenatal diagnosis using recombinant DNA techniques or enzyme assay. Chorionic biopsies were obtained from 40 women undergoing elective first trimester termination of pregnancy (8-12 weeks) under general anaesthesia. The villus tissue was isolated from maternal tissue under a dissection microscope and the presence of any remaining contamination was ascertained by conventional histology and immuno-cytochemical examinations. A high level of success was achieved in obtaining a pure fetal sample. In the first 20 samples the DNA yield obtained using the method of Williamson et al [1] was found to be 0.5 +/- 0.5 micrograms/mg wet weight of villus tissue (mean +/- 1 SD). In the subsequent 20 biopsies using a modified procedure, the yield was significantly improved to 1.0 +/- 0.65 (p less than 0.002). A normal range for the enzyme iduronate sulphatase, which is deficient in Hunter's syndrome (mucopolysaccharidosis II), is also reported. It is suggested that as little as 20 mg of chorionic villi may be used to provide sufficient material for a reliable study using recombinant DNA or biochemical methods.

Amniotic Fluid↗
PubMed1985

Immunocytochemical identification of cytotrophoblast from other mononuclear cell populations isolated from first-trimester human chorionic villi.

Trophoblast biologists are often uncertain as to what cell types they are investigating because the mononuclear cell populations prepared from trypsinization of human first-trimester chorionic villi are morphologically very similar. In the present study, immunocytochemical and phagocytic markers have been used to distinguish cytotrophoblast populations from cell types derived from the mesenchyme of the chorionic villus. Two anti-trophoblast monoclonal antibodies generated in our own laboratory (18B/A5 and 18A/C4) were found to be very efficient in identifying cytotrophoblast, which made up 35-40% of the cells in a smear. Most cytotrophoblast cells did not stain with a monoclonal anti-HLA-A,B,C antibody but a few cells (5%) were found to express both trophoblast and HLA-A,B,C antigens by a double-labelling technique. Endothelial cells from villous capillaries could be identified by a rabbit anti-factor VIII antibody. These cells formed 28% of the population in a cytospin smear. Macrophages from the villous mesenchyme were less readily separable as neither specific monoclonal antibodies nor localization of enzymes were found to be effective. However, these cells could be identified by their ability to phagocytose carmine. About 15% of the cells in a smear consisted of macrophages. The procedure described should prove useful in judging the efficiency of isolation methods from human placental cells.

Antibodies, Monoclonal↗