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PubMed1990

Cytotoxic antibody against trophoblast and lymphocytes present in pregnancy with intrauterine 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.

Adult↗
PubMed1993

Effect of IFN-gamma and IFN-alpha on killing of human trophoblast by decidual LAK cells.

Human decidual large granular lymphocytes are capable of killing both normal trophoblast and choriocarcinoma cells after stimulation with recombinant interleukin-2 (rIL-2) in vitro. We now show that pre-treatment of normal trophoblast and JEG-3 choriocarcinoma cells with interferon-gamma (IFN-gamma) protects these target cells from killing by decidual effectors, although pre-treatment with interferon-alpha (IFN-alpha) had no such effect. In contrast, JAR choriocarcinoma cells were readily killed before and after exposure to both IFN-gamma and IFN-alpha. This protective effect might be due to upregulation of non-classical trophoblast HLA Class I molecules, as IFN-gamma but not IFN-alpha enhances Class I surface expression in both normal trophoblast and JEG-3 choriocarcinoma cells. JAR cells, however, are constitutively and inductively negative for Class I expression.

Cell Death↗
PubMed1993

Adhesion of lymphocytic cells to human trophoblast cells in vitro.

The adherence of lymphocytic MOLT-4/clone 8 cells and normal human peripheral blood mononuclear cells (PBMCs) to primary cultures of term human syncytiotrophoblast has been characterized. Adherence was measured using a fluorescence-based assay in which leukocytic cells were labelled with calcein-AM. Adherence of MOLT cells to syncytiotrophoblast increased in a time-dependent fashion up to about 4 h after which adhesion decreased. Adhesion was detectable at 4 degrees C but was greatly reduced compared to that seen at 37 degrees C. Binding increased linearly as the ratio of MOLT cells to trophoblast was increased. Scanning and transmission electron microscopy of MOLT cell-trophoblast cocultures revealed lymphocytes adherent to the free microvillous surface of the syncytiotrophoblast masses. MOLT cells also adhered to cytotrophoblast but the extent of binding was lower than to syncytiotrophoblast. Normal human peripheral blood mononuclear cells adhered to syncytiotrophoblast. Preincubation of trophoblast cells with trypsin in the presence of calcium had no effect on subsequent adhesion of MOLT cells. However, preincubation of trophoblast cells with trypsin in the absence of divalent cations reduced subsequent adhesion. Adhesion of MOLT cells to syncytiotrophoblast was dependent on magnesium and calcium. These results show for the first time that lymphocytic cells adhere to isolated human syncytiotrophoblast and raise the possibility that this may be an important phenomenon in vivo.

Calcium↗
PubMed1994

Decidual-trophoblast interactions: decidual lymphoid cell function in normal, anembryonic, missed abortion and ectopic human pregnancy.

This study was designed to investigate the consequences of decidua-trophoblast interactions on the phenotype, spontaneous and induced proliferation and immunoregulatory potential of decidual leukocytes in normal pregnancies (NP), anembryonic pregnancies (AP), missed abortions (MA) and ectopic pregnancies (EP). Spontaneous proliferation of decidual non-adherent cells (NAD) from pregnancies with viable trophoblast inside the uterus is significantly higher than proliferation of peripheral blood lymphocytes (PBL) from the same groups (P < 0.001 for NP; P < 0.05 for AP). Spontaneous proliferation of decidual NAD cells from NP was higher (P < 0.001) when compared with AP and EP. The induced (PHA and Con A) responses of PBL from women with normal and pathological pregnancies were significantly higher than that of decidual NAD cells (P < 0.001). Higher proliferation of NAD decidual cells was obtained when Con A-stimulated NP were compared with MA and EP (P < 0.01). The interaction of viable trophoblast with intrauterine decidua appears to be a prerequisite for the activation of NAD suppressor cells, since NAD cells from MA produced stimulation instead of suppression, and NAD cells from EP had no suppressive effect. On the contrary, both NAD and adherent (AD) decidual leukocytes from NP and AP produced very strong suppression of PHA or alloantigen-induced PBL proliferation. The contact between trophoblast and AD decidual leukocytes is not necessary for their suppressive function, since even higher suppression is obtained with the cells from ectopic pregnancies.

Abortion, Missed↗
PubMed1995

Cytokine regulation of HLA-G expression in human trophoblast cell lines.

HLA class I genes are differentially expressed among subpopulations of cells in first trimester human placentas. In this study, HLA class I protein was detected in extravillous cytotrophoblast cells by immunohistochemistry using the monoclonal antibody W6/32. In the same trophoblast subpopulation, class Ib proteins were identified with two monoclonal antibodies, 87G (anti-HLA-G) and 131 (anti-HLA-A/G) and class Ia protein was detected with the monoclonal antibody, 4E (anti-HLA-B/C). All of the antibodies also identified antigens on the human trophoblast-derived choriocarcinoma cell line, JEG-3. Therefore, the JEG-3 cells were used as a model system to study cytokine regulation of HLA-G in trophoblast cells. Northern blot hybridization studies showed that interferons (IFN-alpha, IFN-beta, IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) modestly enhanced steady state levels of HLA-G mRNA. Yet analysis of HLA-G protein by immunocytochemistry and flow cytometry failed to identify any changes in intracellular or membrane expression of HLA-G protein following cytokine treatment. Resistance to upregulation of HLA class I antigens was not a general feature of JEG-3 cells; IFNs enhanced expression of HLA-B/C as well as HLA class I light chain, beta 2-microglobulin. HLA null Jar choriocarcinoma cells did not contain HLA-G mRNA or antigen and exposure to cytokines had no effect on HLA-G. The results of this study are consistent with the postulate that trophoblast cell expression of HLA-G is stringently regulated and is controlled in part by post-transcriptional mechanisms.

Cell Line↗
PubMed1993

In situ hybridization and northern blot demonstration of HLA-G mRNA in human trophoblast populations by locus-specific oligonucleotide.

Trophoblast cells from normal first trimester pregnancies have been shown to express the nonclassical class I molecule, HLA-G, which is nonpolymorphic and has a heavy chain of 40 kD. In this study, in situ hybridization experiments were performed using a biotinylated HLA-G specific oligonucleotide on sections of normal placenta including the implantation site. HLA-G mRNA was identified in all extravillous trophoblast populations including the cytotrophoblast cell columns, interstitial trophoblast, endovascular trophoblast, and placental bed giant cells. In addition, villous cytotrophoblast and villous mesenchymal cells also contained HLA-G transcripts, but villous syncytiotrophoblast was always negative.

Blotting, Northern↗
PubMed1995

Glycoprotein and ganglioside changes in human trophoblasts after exposure to pulsed Doppler ultrasound.

Changes in glycoprotein and ganglioside composition in human trophoblasts (eighth week of gestation) after in vitro exposure to pulsed Doppler ultrasound (pulse duration 1.22 microseconds; repetition frequency 11.1 kHz; center frequency 4 MHz; ISPPA = 175.5 W/cm2; ISPTA = 0.59 W/cm2) were investigated. Evacuated trophoblasts were divided in two halves and insonated for 10 min on top of a 6-cm layer of 5% gelatin in 50-mL tubes (Falcon) at 37 degrees C. One half of each trophoblast was sham insonated and served as an internal control. After insonation trophoblasts were maintained at 37 degrees C for 24 h. Glycoproteins were detected using alpha-D-mannose specific lectins from Galanthus nivalis and Narcissus pseudonarcissus. A decrease in the expression of mannose containing glycoprotein mgp47 and an increase in expression of mgp54 were observed. Ganglioside composition was also significantly altered. Concentrations of two gangliosides migrating similarly to GM2, and one similarly to GQ1, decreased by more than 75%. At the same time, concentrations of one ganglioside migrating similarly to GM3, and two other unidentified gangliosides increased two- to fourfold.

Chromatography, Thin Layer↗
PubMed1992

Effects of calcitonin on human trophoblastic cells in culture: absence of autocrine control.

The purpose of this work was to investigate the effects of calcitonin (CT) on trophoblastic cells with respect to cAMP levels and human chorionic gonadotrophin (hCG) secretion in cultured cells from first-trimester and term placentas and in a choriocarcinoma cell line (JEG-3). The expression of the CT gene was investigated to elucidate a putative autocrine control of CT during pregnancy. The addition of salmon CT (10(-10) M and above) resulted in concentration-dependent increases in cAMP secretion by normal trophoblastic cells from term and first-trimester placentas. Moreover, CT was found to increase cAMP secretion preferentially in completely differentiated cells, i.e. after 4-7 days in culture. Addition to the culture medium of JEG-3 cells slightly increased cAMP secretion only at a concentration of 10(-8) M. The basal level of hCG in the medium was found to be higher in the first-trimester than in the term trophoblast culture, but salmon CT induced an increase in hCG secretion by term placenta cells only. CT gene expression in our experimental model was investigated to elucidate a putative autocrine control of CT action during pregnancy. It was not found to be expressed in syncytiotrophoblast cells from either first-trimester or term placenta cells by the method used. Our data demonstrate the absence of autocrine control of CT effects in trophoblastic cells, and suggest that CT is likely to exert its effect preferentially on differentiated cells.

Blotting, Northern↗
PubMed2005

Trophoblast cells of ruminant placentas--A minireview.

The ruminant placenta is classified as cotyledonary and synepitheliochorial on the basis of its gross anatomical features and histological characteristics, respectively. The richly vascularized embryonic chorioallantois is lined on its external surface by cells of the trophectodermal epithelium. These cells which assume specialized functions are referred to as trophoblast cells. Two morphologically and functionally distinct cell types have been recognized in the trophectoderm of the placenta of ruminant animals. These are the mononucleate trophoblast cells and the binucleate trophoblast cells. The occurrence, morphological characteristics, and specialized functions of these trophoblast cells, in relation to conceptus nutrition and survival in utero are discussed in this review.

Animals↗
PubMed2006

Contribution of inducible nitric oxide synthase and cyclooxygenase-2 to apoptosis induction in smooth chorion trophoblast cells of human fetal membrane tissues.

We examined the contribution of apoptosis- and oxidative stress-associated genes to apoptosis induction in trophoblast cells of human fetal membrane tissues undergoing apoptosis during in vitro incubation. RT-PCR analyses demonstrated an increased level of HO-1, Mn-SOD, Cox-2, iNOS, TNFalpha, TNFR1, IL-1beta, IL-6, Bax, Bak, and Bad gene expression, while Bcl-2 mRNA expression level decreased. Western blot analyses demonstrated an increase in iNOS, Cox-2, and HO-1 protein levels; a decrease in pro-caspase-3 and 9, proform-PARP, and Apaf-1 protein levels; a leakage of cytochrome c from the mitochondria. An antioxidative reagent, general and selective Cox-2 inhibitors, and an iNOS inhibitor suppressed in vitro progression of the apoptosis. Furthermore, an NO donor reagent induced apoptosis in primary cultured trophoblast cells. Therefore, we concluded that the induction of apoptosis in the smooth chorion trophoblasts is mediated through oxidative stress induction followed by mitochondria damage, suggesting that iNOS and Cox-2 play an important role in the apoptosis induction in trophoblasts of human fetal membrane tissues.

Apoptosis↗
PubMed2006

Placental trophoblast inclusions in autism spectrum disorder.

BACKGROUND: Microscopic examination of placental tissue may provide a route to assessing risk and understanding underlying biology of autism. METHODS: Occurrence of a distinctive microscopic placental morphological abnormality, the trophoblast inclusion, was assessed using archived placental tissue. The rate of occurrence of trophoblast inclusion-positive slides observed for 13 individuals with autism spectrum disorder (ASD) was compared to the rate in an anonymous consecutive birth cohort. RESULTS: The occurrence of inclusion positive slides was significantly greater in the ASD group compared to the control group (6/27 slides, 22.2% vs. 12/154, 7.8%; Fisher Exact Test, two-tailed p = .033; relative risk 2.85). The proportion of positive cases was also greater in the ASD group (5/13 cases, 38.5% vs. 8/61, 13.1%; Fisher Exact, two-tailed p = .044; relative risk 2.93). Behavioral severity scores did not differ across groups of inclusion positive (N = 4) and negative (N = 8) ASD individuals. CONCLUSIONS: Although probably not functionally detrimental or causative, the greater occurrence of placental trophoblast inclusions observed in ASD individuals may reflect altered early developmental processes. Further research is required to replicate the basic finding, to understand the basis for the trophoblastic abnormality, and to determine the utility of the measure in early detection of ASD.

Adolescent↗
PubMed2005

Cell reproduction and genome multiplication in the proliferative and invasive trophoblast cell populations of mammalian placenta.

Spatiotemporal "time-table" of ways of cell reproduction (mitosis, restitutional mitosis, endomitosis, endoreduplication) of trophoblast cell populations is described. The populations of mitotically active trophoblast cells (diploid and low-polyploid) are located mostly out of contact with maternal tissues. In rodent placenta they mainly switch from mitotic cycle to polyploidizing (restitutional) mitoses and reach 4c-8c. Thereafter they switch to endoreduplication and reach 16c-64c. Following a series of endoreduplication cycles a part of this cell population sets apart and penetrates deeply into the decidualized endometrium and myometrium, their capabilities for replication being lost progressively (in rodent--256c-1024c). The invasive trophoblast cells that reach 256c-1024c via endoreduplication simultaneously form a barrier between semiallogenic fetal and maternal tissues. Arrest of mitoses and complete repression of DNA replication after a series of endoreduplication cycles makes hardly probable the renewal of mitotic activity in the deeply invading tertiary giant trophoblast cells, thereby preventing the possibility of their ectopic expanding in the maternal tissues during the normal pregnancy.

Animals↗
PubMed2005

T-helper 2 and 3 type immunity to trophoblast in successful in vitro fertilization-embryo transfer.

OBJECTIVE: To determine whether peripheral blood mononuclear cell (PBMC) secretion of T-helper (Th)-1 type cytokines and Th-2 and Th-3 type cytokines in women undergoing in vitro fertilization-embryo transfer (IVF-ET) is associated with therapeutic failure and success, respectively. DESIGN: Cohort study. SETTING: Academic medical center. PATIENT(S): One hundred one women undergoing IVF-ET and 19 fertile controls. INTERVENTION(S): Peripheral blood was obtained from women undergoing IVF-ET before oocyte retrieval and from 19 nonpregnant fertile controls. The PBMCs were cultured in the presence or absence of a protein extract from either a trophoblast cell line or sperm membrane. MAIN OUTCOME MEASURE(S): Supernatants from PBMC cultures were tested by enzyme-linked immunoabsorbent assay (ELISA) for the Th-1 type cytokines tumor necrosis factor-alpha (TNF)-alpha) and interferon-gamma (IFN)-gamma), the Th-2 type cytokines interleukin (IL)-6 and IL-10, and the Th-3 type cytokine transforming growth factor (TGF)-beta1. RESULT(S): Levels of IL-6 and IL-10 were significantly higher in controls than in infertile women with endometriosis, and levels of IL-10 were higher in controls than in women with unexplained infertility. No differences were found in unstimulated levels of TNF-alpha, IFN-gamma, or TGF-beta1 between infertile patients and controls. In trophoblast-stimulated PBMC cultures, levels of TGF-beta1 were significantly lower in subjects who experienced failed compared with ongoing pregnancies. CONCLUSION(S): Baseline PBMC secretion of IL-6 and IL-10 is higher in fertile controls than in women with endometriosis, and IL-10 secretion is also higher than in women with unexplained infertility. Trophoblast-stimulated PBMC secretion of TGF-beta1 is positively associated with the establishment of successful pregnancy in women undergoing IVF-ET. Our study provides novel evidence to support a faciliatory role of Th-2 and Th-3 type responses to trophoblast in early pregnancy.

Analysis of Variance↗
PubMedDate not supplied

Uterine epithelioid trophoblastic tumour in a red-tailed guenon (Cercopithecus ascanius).

Epithelioid trophoblastic tumour (ETT), a rare neoplasm of chorionic-type intermediate trophoblastic cells in the human female, was diagnosed in the uterus of a red-tailed guenon, a non-human primate. The animal, having had two live births, had a recent history of heavy vaginal bleeding. Four years after the last known pregnancy, the animal developed a large invasive mass involving the uterus, right ovary and abdominal wall. The tumour was removed surgically, but at necropsy 1.5 years later was found to have a recurrent neoplasm. Histologically, the original mass consisted of nests and cords of mononuclear intermediate trophoblastic cells whose borders were accentuated by intimately associated eosinophilic hyaline extracellular proteinaceous material. Extensive coalescing areas of necrosis with mineralization surrounding islands of viable neoplastic cells created a "geographical" pattern of necrosis. Immunohistochemical examination revealed that neoplastic cells were diffusely strongly positive for cytokeratin 18, and focally positive for human placental lactogen. The histopathological and immunolabelling patterns were consistent with ETT in human beings. This is the first reported case of epithelioid trophoblastic tumour in a non-human species.

Animals↗
PubMed2004

Modulation of ezrin and E-cadherin expression by IL-1beta and TGF-beta1 in human trophoblasts.

OBJECTIVES: The present study examines the effects of IL-1beta and TGF-beta1 in modulation of ezrin, E-cadherin, CD44 and beta-catenin expression in human trophoblast cells which may lead to their altered cytoskeleton dynamics during cell-to-cell and cell-to-matrix interactions. METHODS: Trophoblast (extravillous and villous) cells isolated and purified from early and term placentae and human choriocarcinoma cell line JEG-3 used in this study were challenged with either IL-1beta or TGF-beta1 (10 ng/ml) for 12 h following which RT-PCR was performed for ezrin, E-cadherin, CD44 and beta-catenin. Immunolocalization of these proteins was carried out in the chorionic villi as well as in the cultured cells stimulated by the cytokines. Western Blot was also performed to study the regulation of ezrin and E-cadherin in primary extravillous, villous and term trophoblast by these cytokines. Scanning electron microscopy (SEM) and Matrigel Invasion Assay was used to study the effect of these cytokines on cellular morphology and invasion. RESULTS: IL-1beta induced a down regulation in the expression of ezrin, E-cadherin and beta-catenin while upregulation of CD44 message in both primary trophoblast and JEG-3 cells. On the contrary, TGF-beta1 exhibited just an opposite effect, i.e. up regulation of ezrin, E-cadherin, beta-catenin, and down regulation of CD44. These observations were further corroborated with the immunolocalization findings of the above proteins in first trimester and term villous tissue, the former having predominance of IL-1beta and the latter of TGF-beta1 [Am. J. Reprod. Immunol. 48 (2002) 210]. Cellular morphology as observed through SEM revealed an enhanced cell-to-matrix adhesion with poor cell-cell interaction following IL-1beta challenge and a strong intercellular adhesion with weak cell-to-matrix interaction in presence of TGF-beta1. Crystal violet staining and Matrigel invasion revealed a higher invasion index following IL-1beta challenge and a low invasion index following TGF-beta1 challenge. CONCLUSION: IL-1beta mediated increased cell-to-matrix interaction with reduced cell-to-cell adhesion along with reduced ezrin and E-cadherin expression is associated with enhanced invasiveness while TGF-beta1 mediated up regulation of cell-to-cell adhesion with reduced cell-to-matrix interaction along with an increased ezrin and E-cadherin expression, is associated with reduced invasiveness, along with an altered cellular morphology. These facts therefore indicate the possible role of the two cytokines during cell motility and invasion through alteration of cell-matrix and cell-cell interaction.

Cadherins↗
PubMed2006

Identification of differences in gene expression in primary cell cultures of human endometrial epithelial cells and trophoblast cells following their interaction.

BACKGROUND: The interaction between epithelial cells of endometrium and trophoblast cells during implantation is presumed to be accompanied by a change in gene expression in the cell types involved. The objective of this study was to identify such differentially expressed genes. METHODS: The interaction between the cell types was simulated in vitro by growing primary cell cultures of human endometrial epithelial cells and trophoblast cells together (co-culture) and separately (control cultures). Gene expression in the cell cultures was compared using the Differential Display method and confirmed using a modified Northern Blot method. RESULTS: Twelve transcripts were identified as being differentially expressed following the interaction between trophoblast and endometrial cells. Some of these sequences show homology to known human genes while other sequences are coding for potential novel genes: (1) one sequence was homologous to the to Homer 1 gene, (2) one identical to the mRNA for XP-G factor, (3) one similar to a hypothetical protein, (4) transcripts showing homologies to a mRNA coding for a cellular proapoptotic protein, and (5) sequences homologous to regions on human chromosomes 5 and 16. Besides, some differentially expressed transcripts have sequences, which could be translated into ribosomal proteins or possibly code for novel proteins. CONCLUSION: These sequences may be important to the course of events following the interaction between endometrial epithelial and trophoblast cells and responsible for implantation.

Blotting, Northern↗
PubMed2005

Persistent gestational trophoblastic disease is rarely, if ever, derived from non-molar first-trimester miscarriage.

Traditional epidemiologic data suggest that persistent gestational trophoblastic disease (pGTD), may follow, and be derived from, either molar pregnancy, non-molar term pregnancy or first-trimester non-molar miscarriage. We examined a database of cases of possible or probable hydatidiform moles and proven pGTD derived from the Regional Trophoblastic Disease Unit, Charing Cross Hospital, London. There were 424 cases (6%), in whom the initial registered diagnosis was that of PHM or CHM but central histopathological review diagnosed a definite non-molar hydropic abortion (HA). In eight of the 424 (2%), although the histology of the most recent index pregnancy was that of non-molar miscarriage, there was a previous history of pregnancy affected by hydatidiform mole; two of these developed subsequent pGTD. Of a further 86 cases referred for a histopathological opinion prior to registration which demonstrated definite non-molar HA, none developed pGTD (zero of 510 (0%, 95% CI 0-0.7%)). During the same period there were 352 cases with pGTD requiring chemotherapy. In 31 cases, the only known pregnancy was the preceding apparent non-molar HA. However, of these, there were only three cases in whom the preceding histological products of conception had been centrally reviewed and were suggestive of non-molar pregnancy. However, in all three of these cases, the specimens were inadequate for definite exclusion of molar pregnancy. In one case in whom no material was available for review, molecular genetic analysis using restriction fragment length polymorphisms was carried out, and the choriocarcinoma was genetically derived from a previous molar pregnancy rather than the preceding HA. There were therefore no cases identified on the database of the trophoblastic disease unit of pGTD requiring treatment in whom the trophoblastic tumour could be genetically proven to have arisen from the preceding first trimester non-molar HA. We suggest that the risk of pGTD developing from a histologically confirmed non-molar HA is less than 1 in 50,000 and that the majority of pGTD cases previously reported to have been caused by a non-molar miscarriage probably represent disease due to an unrecognised early molar pregnancy.

Abortion, Spontaneous↗
PubMed2004

Early trophoblast determination and stem cell maintenance in the mouse--a review.

The first priority of a mammalian embryo is to establish an intimate relationship with its mother. This is accomplished by precocious differentiation of the trophoblast lineage, which mediates uterine implantation and initiates the process of placentation. Surprisingly little is known about the molecular mechanisms that drive trophectoderm differentiation from the equipotent blastomeres of the morula. Somewhat more is known about the maintenance of trophoblast stem cells, once this lineage has been established. The first half of this review will focus on determination of the mouse trophoblast lineage and the second half will discuss the maintenance of trophoblast stem cells.

Animals↗