bromochloroacetic-acid and Abortion--Spontaneous

To examine immune cell phenotypes in viable tubal pregnancies (VTP) and in tubal abortions (TA).. Paraffin-embedded specimens of VTP (n=7) and ongoing TA (n=6) were double-stained for cytokeratin for trophoblast as well as for CD45, CD3, CD8, CD68 and CD20 for immune cell phenotypes. In all cases, the amniotic sac was detected by ultrasound. Histological examination showed no evidence of necrosis within the tissues included in this study. Quantification of the subpopulations was performed in each slide by two independent examiners in five areas (0.085 mm2 each) of the invasion zone as marked by cytokeratin-positive stromal extravillous trophoblast (EVT) cells. For statistical analysis, the non-parametric two-tailed t-test was used (p<0.05).. The differences in the number of CD45(+), CD68(+) and CD20(+) cells was significant (p=0.0423, p=0.0469 and p=0.0494, respectively); however, the number of CD3(+), and among those the number of CD8(+) cells, was approximately eight-fold higher in TA than in VTP (p<0.0001 and p=0.0012, respectively).. The unequal distribution of CD8(+) cells in VTP and TA suggests a significant role of this immune cell phenotype in the further outcome of a tubal pregnancy either to an abortive or a viable, potentially life-threatening, entity.

Topics: Abortion, Spontaneous; Adult; Amnion; Antigens, CD; CD8-Positive T-Lymphocytes; Female; Humans; Keratins; Pregnancy; Trophoblasts

Indoleamine 2,3-dioxygenase (IDO) expression in fetal trophoblast and decidual antigen-presenting cells has been proposed to inactivate maternal T cells and thereby prevent rejection of the "fetal allograft" in early pregnancy. Psychic stress has been proposed to cause miscarriages as well as infertility, at the same time in pregnancy when blockade of IDO causes loss, but the suggested mechanism of stress-triggered loss has been an increased ratio of pro-rejection Th1-type cytokines to anti-rejection Th2/3 cytokines. Could stress act by reducing IDO expression?. Using DBA/2-mated A/J mice where stress causes early pregnancy failure, we examined the role of stress in reducing IDO versus increasing Th1/Th2 ratio in deciduas. IDO loss was also examined in human decidua associated with pregnancy failure.. A post-implantation sonic stress increased the pregnancy failure rate, increased the Th1/Th2 ratio, but did not reduce IDO. IDO was reduced, and Th1/Th2 ratios increased in A/J mice pre-immunized against paternal DBA/2 antigens, and concomitant stress increased these effects. The rate of pregnancy failure was not further increased consistent with recent discoveries of factors that limit the impact of Th1 cytokines at the feto-maternal interface. In deciduas from spontaneous miscarriage patients, IDO(+) cell frequencies were low in only 30% of patients. CD3(+) T-cell numbers and percentage terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end-labelling (TUNEL)(+) apoptotic T cells were increased, but the level of IDO did not correlate with likelihood of apoptosis.. Loss of an allogeneic embryo in early pregnancy is more likely to be due to a high Th1/Th2 ratio than loss of putative protection by IDO.

Topics: Abortion, Induced; Abortion, Spontaneous; Animals; Cytokines; Female; Humans; Immunohistochemistry; Indoleamine-Pyrrole 2,3,-Dioxygenase; Keratins; Lymphocyte Count; Male; Mice; Mice, Inbred A; Mice, Inbred DBA; Pregnancy; Th1 Cells; Th2 Cells; Uterus

Cytokeratin 7 (CK7) is currently regarded as the best marker for trophoblast cells, while CD200 (OX-2), known as 'tolerance signal', plays an important role in normal pregnancy. In this study, the status of CD200 expression was investigated in BALB/c x C57BL/6 and BALB/c x BALB/c mating combinations designed as allogeneic and syngeneic murine models of induced embryo resorption, in which the resorption rate was boosted by an i.p. injection of poly (I:C), a synthetic double-stranded RNA. The percentage of CD200+ cells in the CK7+ cell population (CD200+ CK7+ percentage) and the absolute number of these cells were determined with flow cytometry, using trophoblast cells collected at day 8.5 and day 13.5 of gestation. The potential effect of poly (I:C) on CD200 expression was also evaluated by detecting the CD200+ CK7+ percentage in trophoblast cells incubated in the presence or absence of poly (I:C), in vitro. The distribution pattern of CD200+ cells at the feto-maternal interface was evaluated by immunocytochemical examination. When 10(4) cells were analyzed at day 8.5 of gestation in each case, no significant difference was observed between the poly (I:C)-treated group and the control PBS group either in the CD200+ CK7+ percentage or in the absolute number of these cells. Similar results were observed both in BALB/c x C57BL/6 mice and in BALB/c x BALB/c mice. However, the CD200+ CK7+ percentage was significantly decreased in the poly (I:C)-treated group when evaluated at day 13.5 of gestation. Accordingly, a dramatically elevated rate of embryo resorption was observed at this time point of pregnancy after the administration of poly (I:C). In addition, the CD200+ CK7+ percentage was significantly lower in trophoblast cells incubated with poly (I:C) at a certain concentration, in vitro, while histocytochemical examination showed the CD200+ cells mainly scattered in placental tissue adjacent to the interface of the placenta and uterus. This indicates that sufficient expression of the CD200 molecule on CK7+ cells at the feto-maternal interface may be necessary for the maintenance of embryos during pregnancy in this rodent model, while poly (I:C) administration may increase embryo resorption, at least partially via direct inhibition of CD200 expression on CK7+ cells.

Topics: Abortion, Spontaneous; Animals; Antigens, CD; Biomarkers; Cell Count; Cells, Cultured; Embryo Loss; Female; Immunohistochemistry; Interferon Inducers; Keratin-7; Keratins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Models, Animal; Placenta; Poly I-C; Pregnancy; Trophoblasts

We previously reported a diminished expression of the heme-degrading enzymes heme oxygenases (HO)-1 and HO-2 in decidua and placenta from mice undergoing Th1-mediated abortion, strongly indicating the protective effect of HO in murine pregnancy maintenance. Here we investigated whether the expression of HO-1 and HO-2 is also reduced at the feto-maternal interface of pathologic human pregnancies.. Immunohistochemistry was used to detect HOs expression in placental and decidual first-trimester tissue from patients with: spontaneous abortion (n = 14), choriocarcinoma (n = 14), hydatidiform mole (H-mole) (n = 12), compared with normally progressing pregnancies (n = 15). Further, we investigated early third-trimester decidual and placental tissue from patients with pre-eclampsia (n = 13) compared with fetal growth retardation (n = 14) as age-matched controls.. In first trimester tissue, we observed a significant reduction of HO-2 expression in invasive trophoblast cells, endothelial cells, and syncytiotrophoblasts in samples from patients with spontaneous abortion compared with normal pregnancy. H-mole samples showed a diminished expression of HO-2 in invasive trophoblast cells and endothelial cells in comparison with NP, whereas choriocarcinoma samples showed no significant differences compared with the control. In third trimester tissue, HO-2 was also reduced in syncytiotrophoblasts and invasive trophoblast cells from pre-eclampsia compared with samples from fetal growth retardation. HO-1 expression was diminished in all pathologies investigated; however, the differences did not reach levels of significance.. Our data indicate that HOs play a crucial role in pregnancy and low expression of HO-2, as observed in pathologic pregnancies, may lead to enhanced levels of free heme at the feto-maternal interface, with subsequent upregulation of adhesion molecules, allowing enhanced inflammatory cells migration to the feto-maternal interface.

Topics: Abortion, Spontaneous; Adult; Choriocarcinoma; Decidua; Down-Regulation; Endothelial Cells; Female; Fetal Growth Retardation; Heme Oxygenase (Decyclizing); Heme Oxygenase-1; Humans; Hydatidiform Mole; Immunohistochemistry; Keratins; Membrane Proteins; Placenta; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Trophoblasts

The objective of this study was to assess apoptotic activity in gestational trophoblastic disease (GTD) and its prognostic value in hydatidiform mole (HM).. Expression of the specific caspase cleavage site within cytokeratin 18 was assessed immunohistochemically using the monoclonal antibody M30 CytoDeath in 12 spontaneous abortions, 22 partial and 57 complete HM, eight choriocarcinoma (CCA) and 28 normal placentas. The M30 immunoreactivity occurred predominantly in the syncytiotrophoblasts. A significantly higher M30 index in HM and CCA was found when compared with normal placentas and spontaneous abortions (P < 0.001). The M30 index of those HM which spontaneously regressed was significantly higher than those HM which developed persistent disease requiring chemotherapy (P < 0.001). The M30 index correlated with another apoptotic index previously detected by TdT-mediated dUTP nick-end labelling (TUNEL) (P = 0.007) and the proliferation index assessed by the Ki67 antigen (P = 0.034).. We conclude that apoptosis is important in the pathogenesis of GTD. Assessment of apoptotic activity in HM by the M30 index may be considered as an alternative prognostic indicator for predicting the clinical behaviour.

Topics: Abortion, Spontaneous; Apoptosis; Caspases; Choriocarcinoma; Female; Follow-Up Studies; Humans; Hydatidiform Mole; In Situ Nick-End Labeling; Keratins; Pregnancy; Prognosis; Trophoblastic Neoplasms; Trophoblastic Tumor, Placental Site; Uterine Neoplasms

Programmed cell death by apoptosis occurs in both fetal and maternal tissues during early pregnancy. To investigate a role for apoptosis at the maternal-fetal interface, we have immunolocalized the bcl-2 protein in formalin-fixed decidual and placental tissue collected from women undergoing surgical termination of pregnancy (n = 22), from women undergoing a sporadic miscarriage (n = 16) and from women with a history of recurrent pregnancy loss (more than three consecutive pregnancy losses; n = 22) undergoing a further miscarriage. In all three groups, bcl-2+ cells were found in aggregates and dispersed in the stroma, and immunoreactivity was observed in glandular epithelium. Double immunostaining revealed that a majority of stromal bcl-2+ cells were CD56+ large granular lymphocytes. A computerized image analysis revealed no significant differences in percentage area of bcl-2 or CD56+ immunostaining. Significantly more biopsies from the surgical termination group (4/10) had > 20% positive immunostaining for CD56 compared with 0% in the other two groups combined (0/20; P < 0.05). Bcl-2 immunoreactivity was observed in the villi syncytiotrophoblast, and staining intensity was consistently greater in the surgical termination group. The possible roles of bcl-2 at the maternal-fetal interface are discussed.

Topics: Abortion, Habitual; Abortion, Induced; Abortion, Spontaneous; Apoptosis; CD56 Antigen; Decidua; Female; Humans; Immunohistochemistry; Keratins; Placenta; Pregnancy; Proto-Oncogene Proteins c-bcl-2; Stromal Cells; Trophoblasts

With spontaneous abortion the conceptus is often expelled and lost right at the beginning, and uterine curettings then contain only endometrial fragments and clotted blood. Even complete embedding of all available material for histologic examination will not reveal any chorionic villi, and ectopic pregnancy can thus not be excluded. In such cases, the intermediate trophoblast can sometimes still be demonstrated within the endometrial tissue. This highly invasive trophoblast is difficult to identify using conventional staining, but cytokeratin antibodies are reliable markers of this cell type. Using immunohistochemistry, these fetal components could be demonstrated in 27 of 95 specimens (28.5%), proving the intrauterine nature of the aborted pregnancy. In some cases the fetal derivation of intermediate trophoblast was demonstrated by using in situ hybridisation to mark repetitive sequences on the Y-chromosome in the interphase nucleus.

Topics: Abortion, Spontaneous; Adult; Chromosomes, Human, Pair 17; Diagnosis, Differential; Dilatation and Curettage; Endometrium; Female; Humans; Immunoenzyme Techniques; In Situ Hybridization; Keratins; Pregnancy; Pregnancy, Ectopic; Trophoblasts; Y Chromosome