Amniotic immune biomarkers as risk factors in women with different symptoms of threatened late miscarriage.

PROBLEM: To investigate risk factors that can help identify the possibility of pregnancy loss in threatened late miscarriage (TLM) patients with and without spontaneous uterine contractions. METHOD OF STUDY: Amniotic immune biomarkers (IL2ß receptor, IL6, IL8, IL10, IL1ß, and TNFα) were assayed, and "sludge" was assessed. Patients without intrauterine infections were treated and followed up until delivery, and pregnancy outcomes were recorded. The two groups were compared for the differences in biomarker levels and "sludge," and the independent associations of biomarkers, "sludge," and other maternal factors with late miscarriage were investigated. RESULTS: The amniotic levels of IL2ßR, IL8, and TNFα were higher in the group with contractions (P < .05). When considered alone, each of the six biomarkers was significantly associated with late miscarriage in the no-contractions group and four of these (IL8, IL10, IL1ß, and TNFα) in the contractions group (P < .05). Biomarker levels were correlated, and in multivariate Cox regression analysis, there was an independent effect only for IL8 in the no-contractions group (HR = 18.16, 95% CI: 5.75-57.43) and TNFα in the contractions group (HR = 4.11, 95% CI: 1.68-10.08). For patients with contractions, IL10, IL8, and IL1ß were different in those with and without "sludge," but no such difference was seen in the no-contractions group. CONCLUSION: For TLM patients without intrauterine infections, amniotic immune biomarkers differ between patients with different symptoms, not only for their levels but also for the impact of these biomarkers on the risk of late miscarriage. These findings suggest that the symptoms of TLM should be considered in the study of miscarriage risk.

Antiapolipoprotein A-1 Autoantibody Positivity Is Associated with Threatened Abortion.

BACKGROUND: Autoantibodies against apolipoprotein A-1 (anti-ApoA-1 IgG) were demonstrated to be associated with cardiovascular outcomes in several inflammatory diseases. As balanced inflammation is critical for uncomplicated pregnancy, we aimed to investigate the prevalence of anti-ApoA-1 IgG and anti-c-terminal ApoA-1 autoantibodies (Ac-terAA1 IgG) in a cohort of pregnant women and their potential relationship with threatened abortion (TA). METHODS: Between 2012 and 2014, 371 consecutive outpatient pregnant women were included in this study and followed until delivery. Anti-ApoA-1 and anti-Ac-terAA1 IgG were measured by ELISA technique on serum samples collected between the 24th and 26th week of pregnancy. Associations with TA were tested using linear regression analysis and C-statistics. RESULTS: Median age was 34 with a prevalence of the Caucasian ethnicity (90.5%). TA occurred in 10 women (2.7%). C-statistics indicated that anti-ApoA-1 and anti-Ac-terAA1 IgG levels upon study inclusion were predictive of TA (0.73, 95% confidence interval [CI] 0.69-0.78, p < 0.001 and 0.76, 95% CI 0.71-0.80, p < 0.001 and 0.76, 95% CI 0.71-0.80, p < 0.001 and 0.76, 95% CI 0.71-0.80, p < 0.001 and 0.76, 95% CI 0.71-0.80. CONCLUSION: Anti-ApoA-1 and anti-Ac-terAA1 IgG are independently associated with TA during pregnancy with an appealing NPV. The causal biological mechanisms underlying this association as well as the possible clinical relevance of these findings require further investigations.

Uterine Natural Killer Cell and Human Leukocyte Antigen-G1 and Human Leukocyte Antigen-G5 Expression in Vaginal Discharge of Threatened-Abortion Women: A Case-Control Study.

The immunotolerant human leukocyte antigen-G (HLA-G) molecules have a major role in fetal-maternal tolerance during pregnancy. Interaction between these molecules and uterine natural killer (uNK) cells inhibitory receptors prevents NK cell invasion against fetus trophoblast cells. The aim of this study was to evaluate the percentages of uNK cells and HLA-G1 and HLA-G5 isoforms expression in vaginal discharge of threatened-abortion women in comparison with control. In a case-control study, we investigated 30 threatened-abortion women with bleeding or spotting less than 20 weeks of pregnancy as compared to 30 normal pregnant women. uNK cells percentage was assessed by flow cytometry. Furthermore, we evaluated HLA-G1 and HLA-G5 isoforms expression by Real-Time PCR in these groups. The results of this study showed that threatened-abortion women had increased uNK cells and decreased T cells percentage in vaginal discharge in comparison with normal pregnant women (p = 0.01, p = 0.003, resp.). In addition, HLA-G1 isoform had lower expression in threatened-abortion women in comparison with control group (p = 0.0001). The increase of uNK cells level with the decrease of HLA-G expression in vaginal discharge of threatened-abortion pregnant women is an indicator of mother's immune dysregulation. It is concluded that HLA-G expression level with uNK cells percentage can be determined as a diagnostic marker for threatened-abortion women.

Spectrum of antibodies to reproductive hormones in threatened abortion.

The spectrum of antibodies to reproductive hormones and the diagnostic significance of their measurements in threatened abortion during trimester I were studied. Enhanced production of antibodies to hormones was detected by ELISA in patients with threatened abortion (N=44) in comparison with women with normal gestation (N=30). These antibodies were detected more often than antiphospholipid antibodies (p<0.05). Antibodies to chorionic gonadotropin (IgM, IgG) and gonadotropin-releasing hormone (IgG) were associated with threatened abortion. According to ROC analysis, their measurements were diagnostically significant in this pathology (AUC>0.8). Subclasses IgG1 and IgG2 predominated among IgG to chorionic gonadotropin. Presumably, antibodies to chorionic gonadotropin and gonadotropin-releasing hormone could serve as independent factors of threatened abortion risk during trimester I.

Immunologic regulation in pregnancy: from mechanism to therapeutic strategy for immunomodulation.

The immunologic interaction between the fetus and the mother is a paradoxical communication that is regulated by fetal antigen presentation and/or by recognition of and reaction to these antigens by the maternal immune system. There have been significant advances in understanding of abnormalities in the maternal-fetal immunologic relationship in the placental bed that can lead to pregnancy disorders. Moreover, immunologic recognition of pregnancy is vital for the maintenance of gestation, and inadequate recognition of fetal antigens may cause abortion. In this paper, we illustrate the complex immunologic aspects of human reproduction in terms of the role of human leukocyte antigen (HLA), immune cells, cytokines and chemokines, and the balance of immunity in pregnancy. In addition, we review the immunologic processes of human reproduction and the current immunologic therapeutic strategies for pathological disorders of pregnancy.

Progesterone - induced blocking factor (PIBF) and Th(1)/Th(2) cytokine in women with threatened spontaneous abortion.

OBJECTIVE: The aim of this prospective study was to compare serum and urine concentrations of progesterone-induced blocking factor (PIBF) and serum concentrations of anti-inflammatory (IL10) and pro-inflammatory (IL6, TNFalpha, IFNgamma) cytokines of women with threatened spontaneous abortion with normal pregnancy and to evaluate the impact of PIBF on outcome of pregnancy. METHODS: A sample of 30 women with threatened spontaneous abortion (study group) and 20 healthy pregnant women (control group) between 6(th) and 24(th) gestational weeks was studied. Serum and urine PIBF, IL10 and IL6, TNFalpha, IFNgamma cytokine concentrations were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Five (16.7%) pregnancies in the study group ended missed abortion vs. none in the control group (P<0.05). Five (20%) threatened aborters delivered between 24(th) and 37(th) weeks of gestation, whereas two (10%) preterm deliveries occurred in the controls (P>0.05). PIBF concentrations in urine (19.5+/-12.9 ng/mL) and serum (214.4+/-120.6 of patients with threatened abortion were significantly lower than in healthy pregnant women (45.3+/-33.7 ng/mL and 357.3+/-159.9 ng/mL, respectively). Women with threatened abortion had significantly lower serum levels of anti-inflammatory cytokine, but levels of proinflammatory cytokines were higher in this group compared with healthy controls. CONCLUSIONS: Determination of progesteron-induced blocking factor level in body fluids in early pregnancy might be used for the diagnosis and prognosis of threatened abortion.

Syngeneic immune-dependent abortions in mice suggest paternal alloantigen-independent mechanisms.

PROBLEM: Recurrent immune-associated miscarriages in humans are thought to result from maternal immune responses to paternal alloantigens. We investigated the role of paternal alloantigens in a mouse model of immune-dependent abortion. METHOD OF STUDY: Sib-crosses of C57Bl/6J (haplotype b/b) mice heterozygous for a targeted hypomorphic allele of the tbp gene (tbp(deltaN/+)) resulted in selective mid-gestational abortion of 88% of the tbp(deltaN/deltaN) fetuses. In dams lacking mature lymphocytes (rag1-/-), nearly all tbp(deltaN/deltaN) fetuses survived to birth, indicating abortions were immune-dependent. Allogeneic pregnancies bearing tbp(deltaN/deltaN) fetuses were established by either hybridizing the paternal lineage to BALB/cJ (haplotype d/d) and mating hybrid tbp(deltaN/+) sires to haplotype b/b tbp(deltaN/+) C57Bl/6J dams, or by transfer of haplotype b/b zygotes from tbp(deltaN/+)x tbp(deltaN/+) matings into pseudopregnant wild-type CByD2F1/J dams (haplotype d/d). RESULTS: Neither hemizygous paternal allogeneic loci nor homozygous allogeneic loci, including a haplotype-mismatched major histocompatibility complex (MHC), increased abortion frequencies. CONCLUSION: Results suggested that mechanisms for maternal tolerance of paternal alloantigens, including mismatched MHC antigens, were intact in these pregnancies, yet maternal immune-dependent paternal antigen-independent abortion of mutants occurred. These data indicate that, in some cases of immune-mediated abortions, the presence of paternal alloantigens can be coincidental and superfluous to the compromising rejection response.

The detection of anti-beta2-glycoprotein I antibodies is associated with increased risk of pregnancy loss in women with threatened abortion in the first trimester.

OBJECTIVE: This study was designed to evaluate whether the detection of serum antiphospholipid autoantibodies may be useful in predicting pregnancy outcome in women with threatened abortion in the first trimester. STUDY DESIGN: A group of 77 pregnant women of between 8 and 12 weeks' gestation with vaginal bleeding was tested for serum antiphospholipid, lupus anticoagulants, anticardiolipin, antinuclear antibodies, and anti-beta2-glycoprotein I antibodies, and was followed up until the spontaneous end of pregnancy. A control group composed of 15 healthy women with uncomplicated gestation was tested contemporarily for the same antibody panel. RESULTS: Of the 77 patients with threatened abortion, 32 (41.5%) progressed to deliver at term and 45 (58.5%) experienced early pregnancy loss. Among the antibodies evaluated, only anti-beta2-glycoprotein I was significantly more frequent in those women whose pregnancy resulted in spontaneous abortion (22/45, 49%) than in those who progressed to term (6/32, 19%) or in the control group (2/15, 13%; p=0.004). This difference was specific to the IgM isotype (p=0.001). After adjustment by multivariate analysis, the odds ratio for pregnancy loss associated with a positive beta2-glycoprotein I antibody test was 5.18 (p=0.001). CONCLUSION: The detection of anti-beta2-glycoprotein I antibodies is associated with an increased risk of pregnancy loss in women with threatened abortion in the first trimester.

Maternal serum levels of pro-inflammatory cytokines in missed and threatened abortion.

PROBLEM: A study of association between pro-inflammatory cytokines, and missed and threatened abortions with good outcome has been performed. METHOD OF STUDY: The presence of pro-inflammatory cytokines, namely interleukin (IL)-8 and IL-12 and the soluble interleukin-2 receptor (SIL-2R) was investigated in maternal serum of 12 patients with threatened abortion twice (at admission and discharge), 14 patients with missed abortion, 14 women with healthy first-trimester pregnancy, and 14 normal non-pregnant women, using specific enzyme-linked immunosorbent assays. RESULTS: SIL-2R and, in particular, IL-12 was detected with significantly higher levels in missed abortion group compared with all other groups. IL-8 was detected with no significant difference among all the groups studied. CONCLUSIONS: In spite of caution due to the small sizes of the subject samples, these results support a role of the immune system in the first trimester pregnancy and hypothesize that missed abortion may be associated with an enhanced Th1 reactivity, whereas threatened abortion with good outcome resembles the normal pregnancy with a non-enhanced Th1 reactivity.

[Serum levels of CA-125 antigen during the first trimester of pregnancy complications and the risk of miscarriage]. / Ocena stezenia antygenu CA125 w surowicy krwi ciezarnych w i trymestrze ciazy powiklanej zagrazajacym poronieniem.

BACKGROUND: The ultrasound, serum beta-HCG and progesterone titres are widely used to assess the risk of miscarriage at the early stages of pregnancy. Though very useful, they are not considered as satisfactorily accurate predictors of imminent abortion during the first three months of pregnancy. AIM OF STUDY: Prospective evaluation of clinical usefulness of serum CA125 from 4-12th week of pregnancy in a group of women with symptoms of imminent abortion. MATERIAL AND METHODS: A study group were 250 consecutive women with symptoms of imminent abortion. Inclusion criteria were: abdominal pain, spotting, ultrasound picture of the embryo in the uterus, monovular pregnancy, normal ultrasound picture of ovaries, gestational age ranging from 4-12 weeks (assessed on a last menstrual period basis with ultrasound corroboration). 55 women with physiological course of pregnancy made up a control group. CA125 levels was evaluated with Roche Cobas Core CA125 II EIA. RESULTS: In patients with symptoms of imminent abortion, serum CA125 titration in 4 to 12 weeks' gestation seems to be valuable only in woman with vaginal spotting or bleeding.

[Relationship between anticardiolipin antibodies and abortion].

OBJECTIVE: To explore the relationship between anticardiolipin antibodies (ACA) and abortion. METHODS: The level of serum ACA in 93 abortion women (abortion group) and 80 normal pregnant women (control group) was determined by the enzyme linked immunoabsorbent assay. The abortion group included threatened abortion (n = 62), inevitable abortion (n = 21), and missed abortion (n = 10). RESULTS: The positive rate of ACA in the abortion group was significantly higher than that in the control group (P < 0.05). The positive rate of ACA in pregnant women with a history of abortion was significantly higher than that in those women without a history of abortion (P < 0.05). The positive rate of ACA was not significantly different between habitual abortion and spontaneous abortion (P > 0.05). CONCLUSION: Anticardiolipin antibodies may induce abortion, but it is not related to habitual abortion.

Bioactive to immunoreactive ratio of circulating human chorionic gonadotropin as possible evaluation for the prognosis of threatened abortion.

OBJECTIVE: In an effort to evaluate the prognosis of threatened abortion, we established the ratio of serum human chorionic gonadotropin (hCG), as measured by bioassay or radioimmunoassay techniques, of samples from patients with threatened abortion or normal pregnancy. STUDY DESIGN: Peripheral blood samples from patients in their first trimester of pregnancy with threatened abortion (n=24), or normal pregnancy (n=12), were assayed for progesterone (RIA), and for immunoactive (DELFIA) and bioactive (mouse Leydig cell testosterone production assay) hCG. RESULTS: Serum progesterone was not statistically different between the threatened-continuing and the threatened-miscarried groups. The ratio of hCG bioactive/hCG immunoactive (B/I) was significantly lower for the patients of the threatened group that experienced abortion. The B/I ratio for the control and threatened-continuing patients was similar. CONCLUSION: The hCG bioactive/hCG immunoactive ratio could be a good indicator of the prognosis of threatened abortion.

[Preliminary results of the heparin treatment in anticardiolipin and antihistone antibodies seropositive pregnant women]. / Wstepne wyniki leczenia heparyna ciezarnych z obecnoscia przeciwcial antykardiolipinowych i antyhistonowych w surowicy krwi.

The presence of elevated titres of anticardiolipin antibodies (ACA) and antihistone antibodies (AHA) in the blood serum is considered as one of serious reasons of repeated pregnancy losses. According to some reports, heparin significantly improves live birth rates in these cases. The aim of the work is an evaluation of the results of the heparin therapy in pregnant women with elevated titres of ACA and/or AHA in blood and bad obstetric anamnesis, or after sterility treatment. Our material consisted of three groups: the first one was composed of 25 ACA- and/or AHA-seropositive pregnant women 30.0 +/- 4.1 years old with 1-5 early miscarriages in past, the second one of six seropositive patients 31.3 +/- 2.8 years old, actually pregnant after the treatment of unexplained sterility (two of them after assisted reproduction) and, finally, in the third group were placed five pregnant ACA- and AHA-seronegative pregnant women 30.8 +/- 2.2 years old with 2-4 abortions of unexplained etiology in past. The ACA IgA, IgM and IgG and AHA IgG levels in blood sera were determined by ELISA. As a positive titre ACA in the class IgA was considered > 7 APL/ml, in the class IgM > 4 MPL/ml, IgG > 7 GPL/ml and in case of AHA IgG > 25 GPL/ml. The patients were treated by heparin 7500-30,000 units daily during all the pregnancy under the control of kaolin-kephalin time. In the first group, where 53 pregnancies from 56 were miscarried (94.6%), after the heparin therapy in 10 women 9 pregnancies of 11 (81.8%) were terminated by live birth (p < 0.001). One of the patients died three days after cesarean section because of myocardial infarction, probably due to sudden stopping of heparin. In the second group three women after heparin treatment delivered live babies, but three untreated aborted. In the last group only two women treated by heparin delivered live babies and three, despite treatment, miscarried. It should be concluded, that heparin therapy in ACA- and/or AHA-positive pregnant women might be a recommended therapeutic method. In cases of antiphospholipid syndrome a special precaution should be undertaken, when stopping the heparin. It seems, that double assay of ACA and AHA in patients with conception troubles might be usefull.

Human chorionic gonadotropin, thyroid function, and immunological indices in threatened abortion.

OBJECTIVE: To evaluate the role of thyroid hormones in maintaining early pregnancy and to examine the association between thyroid physiological functions and immunological parameters. METHODS: Forty-five pregnant women with a clinical diagnosis of threatened abortion and a live fetus and 30 normal pregnant women were included in the study. Blood samples were taken on admission to the hospital. The patients were divided retrospectively into two groups on the basis of outcome: 1) 31 women who did not miscarry (positive outcome) and 2) 14 women who miscarried (negative outcome). Plasma TSH, free triiodothyronine (fT3), free thyroxine (fT4), hCG, immunoglobulin (Ig) G and IgM concentrations and blood counts were determined in each patient. RESULTS: Human chorionic gonadotropin was significantly higher in women who did not abort (39.4 +/- 16.9 IU/mL) than in women who miscarried (17.6 +/- 14.8 IU/mL, P < .001). Free thyroxine but not fT3 was lower in patients with negative outcome (1.25 +/- 0.26 ng/mL compared with 1.98 +/- 0.22 ng/mL, P < .001) and IgG and IgM plasma levels were higher (780 +/- 500 ng/mL compared with 470 +/- 300 ng/mL and 930 +/- 400 ng/mL compared with 650 +/- 280 ng/mL, respectively, P < .05). Plasma TSH levels were higher in patients with negative outcomes (1.72 +/- 0.84 mIU/mL compared to 1.01 +/- 0.41 mIU/mL, P < .001). Plasma concentrations of hCG and thyroid hormones were significantly correlated with peripheral blood lymphocyte and neutrophil counts only in the group of women who aborted. CONCLUSION: Our results indicate that maternal immune response, trophoblast function, and maternal thyroid function are somehow correlated. The presence of low concentrations of hCG and fT4 and high levels of TSH and gamma globulins in women with threatened abortion suggests a negative outcome for the pregnancy.

[Toxoplasmosis and threatened abortion]. / Toxoplasmosis y aborto en evolución.

To know the situation of the toxoplasmosis in Comitán Chiapas, we made a serological indirect inmunofluorecent antibody test (IFA) to the population of this city and to fifty women with abortion in evolution. The results show us that around five percent of the population in general have positive title of antitoxoplasma gondii antibiodies, and 18% in the women with abortion evolution case. The statistics concluded that seropositive for this parasitic disease is real higher between cases of abortion than population in general (P < 0.006), as well as it is significantly higher in abortion cases than women of the general population of Comitán Chiapas (P < 0.01).

[The evaluation of expression of T-lymphocytes and NK cell surface receptors in women with threatened miscarriage]. / Ocena ekspresji antygenów powierzchniowych limfocytów T i komórek NK u pacjentek w ciazy zagrozonej poronieniem.

OBJECTIVES: Recent studies emphasise an important role of immunological mechanisms in pregnancy maintenance. Therefore, unravelling mechanisms regulating placentogenesis are critical to understanding the pathogenesis of recurrent spontaneous abortion. MATERIALS AND METHODS: We studied 49 women with threatened abortion and 24 healthy pregnant women. In addition, we studied 17 women with the history of recurrent spontaneous abortion and 10 healthy nonpregnant women in reproductive age with the previous successful pregnancy outcome. CD3, CD45 RO, CD4, CD8, CD16 expression on peripheral blood lymphocytes were studied using flow cytometry. RESULTS: We determined that there is significantly higher CD4 expression in pregnant recurrent aborters compared to control (p < 0.05). Pregnant recurrent spontaneous aborters, with the successful pregnancy outcome have significantly lower CD16 expression compared to those, who abort (p < 0.05). CONCLUSION: Our studies may indicate that T cell and NK cell can be involved in the pathogenesis of pregnancy loss.

Serum CA 125 and first trimester abortion.

OBJECTIVE: The study was carried out to assess the clinical value of serum CA 125 in association with serum beta-human chorionic gonadotropin (hCG) for predicting pregnancy outcome. METHODS: One hundred spontaneous pregnancies, 52 non-threatened pregnancies and 48 with threatened abortions, observed in the Department of Obstetrics and Gynecology at the University 'La Sapienza', Rome, Italy, were evaluated during the first trimester using ultrasound examination, CA 125 and beta-hCG titrations. RESULTS: Threatened pregnancies had statistically significantly higher CA 125 serum values than non-threatened pregnancies, especially those with a negative outcome (P < 0.01). The CA 125 levels in the threatened pregnancies were positively correlated with the tropho-decidual hematoma volume (r=0.839, P < 0.0001). The CA 125-beta-hCG association showed a higher prognostic value (sensitivity 78.9%, specificity 96.5%) in assessing pregnancy outcome than CA 125 or 0-hCG alone (sensitivity 78.9% and 57.9%, respectively; specificity 75.8% and 86.2%, respectively). CONCLUSIONS: Our findings are in accordance with the hypothesis of a tropho-decidual origin of this marker, suggesting its possible usefulness in the prognostic evaluation of first trimester threatened abortion.

[The interferon status and cellular immunity of pregnant women with viral hepatitis B]. / Interferonovyi status i kletochnyi immunitet u beremennykh zhenshchin, bol'nykh ostrym virusnym gepatitom B.

The examination of 62 pregnant women with acute viral hepatitis B has established in them inhibition of interferon genesis and the presence of secondary T-cell immunodeficiency. The most pronounced and long-lasting was a fall of gamma-interferon. A simultaneous lowering of alpha-, gamma- and serum interferon was indicative of complicated pregnancy or premorbid condition. A persistent deficient production of gamma-interferon and T lymphocytes, imbalance of their subpopulations promoted long-term course of hepatitis and persistence of HBsAg.

[Antiphospholipid antibodies in women with reproductive disorders]. / Antifosfolipidové protilátky u zen s nekterými poruchami reprodukce.

The authors examined in serum of 121 women, using the ELISA method, anticardiolipine antibodies (ACA) class IgG and IgM. The examined women were divided into a group of women with a normal pregnancy (n = 14), a group of women with imminent abortion during the first trimester (n = 10) and a group of women treated for prolonged periods on account of sterility (n = 97). In women with a normal pregnancy only in one a positive titre in the IgG class was recorded. In the group with imminent abortion a positive ACA titre was found in three patients (30%); in women with negative results of the ACA examination, pregnancy continued. In sterile women positive ACA tests were recorded in 14.5%.

[The hormonal regulation of thymus activity in threatened abortion]. / Hormonal'na rehuliatsiia tymichnoï aktyvnosti pry nedonoshuvanni vahitnosti.

It is stated that in women with the abortion threat at any term of gestation (I, II, III trimesters) the endocrine function of the thymus essentially increases. The role of hormones in the thymic activity disorder is studied. It is shown that insufficiency of the functional activity of the trophoblast ranks first in the mechanism of hormonal regulation of thymic endocrine function in nonbearing of pregnancy. The other significant reason of an essential increase in the titer of thymic serum factor (TFS) is a disorder in interconnections inside the hypophyseal-adrenal system. Determination of TFS and the above hormonal indices can be used as the prognostic test of the pregnancy course.