Peripheral Blood Of Pregnant Women Research Articles

Relationship between amniotic fluid inflammatory factors and pregnancy outcomes after emergency cervical cerclage

Objective: To explore the relationship between amniotic fluid and peripheral blood inflammatory factors and the pregnancy outcomes after emergency cervical cerclage, and to identify effective indicators for predicting adverse pregnancy outcomes after the procedure. Methods: A case-control study was conducted, including pregnant women who were hospitalized at Sun Yat-sen Memorial Hospital, from January 1, 2013, to July 31, 2019, and underwent emergency cervical cerclage due to cervical dilatation at gestational age between 16 and 28 weeks. A total of 85 pregnant women who underwent amniocentesis for the detection of amniotic fluid inflammatory factors during the perioperative period were included. Based on whether their baby was perinatal death, the participants were divided into the case group (28 cases with perinatal death) and the control group (57 cases with live births). Univariate logistic regression analysis was performed to identify risk factors associated with adverse pregnancy outcomes, followed by multivariate logistic regression analysis to establish a regression model and nomogram. Results: (1) The levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6, IL-8, IL-10 in the amniotic fluid during the perioperative period and postoperative serum C-reactive protein (CRP) were significantly higher in the case group compared to the control group (all P<0.05). The case group underwent emergency cervical cerclage at an earlier gestational age compared to the control group, and their cervical dilation was greater than that of the control group (all P<0.05). However, there were no significant differences in the white blood cell counts, neutrophil percentage, and the level of preoperative CRP in the peripheral blood of pregnant women during the perioperative period (all P>0.05). (2) Univariate logistic regression analysis showed that the levels of amniotic fluid WBC, TNF-α, IL-1β, IL-2 receptor (IL-2R), IL-6, IL-8, IL-10, postoperative CRP in the peripheral blood, gestational age at cerclage and cervical dilation were associated with adverse pregnancy outcomes (all P<0.05). Multivariate regression analysis indicated that only the levels of amniotic fluid WBC and TNF-α were independent risk factors for perinatal death. (3) Based on clinical practice, a multivariate logistic regression model was constructed including the levels of amniotic fluid TNF-α, WBC, gestational age at cervical cerclage, and cervical dilation. A nomogram and calibration curve were plotted, which suggested its good predictive value for adverse pregnancy outcomes. Conclusions: During the perioperative period of emergency cervical cerclage, the levels of amniotic fluid WBC, TNF-α, IL-1β, IL-2R, IL-6, IL-8, IL-10 are associated with adverse pregnancy outcomes, with amniotic fluid WBC and TNF-α showing the closest relationship. However, there is no significant correlation between maternal peripheral hemogram during the perioperative period and adverse pregnancy outcomes. A model constructed by amniotic fluid TNF-α, WBC, cervical cerclage gestational age, and cervical dilation has a good predictive effect on adverse pregnancy outcomes.

P-390 The percentage of regulatory T cells is associated with T follicular helper cell quantity in the peripheral blood of pregnant women

Abstract Study question Is there a relationship between the percentage of peripheral regulatory T cells (Treg) and other T cell subtypes in the first trimester of pregnancy? Summary answer The percentage of Treg cells showed a significant positive correlation with the quantity of T follicular helper cells (TFH). What is known already Regulatory T cells (Treg) play an essential role in the process of embryo implantation and maintenance of normal pregnancy. The quantity of these cells in the peripheral blood expands during the first trimester of pregnancy. Several studies report the importance of Treg in the control of the inflammatory process and in the pathophysiology of common adverse pregnancy outcomes. However, there is a scarce data about the quantitative relation between Treg and other important T sub-types during pregnancy. Study design, size, duration This is an observational study of 93 women, 28 to 53 years of age (mean 38 years), who had a blood sample during the first trimester (at 8-10 weeks of pregnancy). The study was conducted between November 2022 and October 2023. The exclusion criteria were history of recent inflammatory or infection disorders, recent antibiotic treatment, autoimmune diseases, oncological diseases, moderate or severe endometriosis, and endometrial polyps. Written informed consent was obtained from each participant. Participants/materials, setting, methods SIx peripheral T cell subtypes were measured by flow cytometry: T helpers % (CD3+CD4+CD8-/CD3+), T killers % (CD3+CD4-CD8+/CD3+), double-positive T cells % (CD3+CD4+CD8+/CD3+), double-negative T cells % (CD3+CD4-CD8-/ CD3+), T follicular helper cells % (CD3+CD4+CD8-CD185+/CD4+), T reg cells (CD4+CD8-CD25+CD127−/CD4+). Spearman’s correlation test was performed with all studied T cell subtypes in SPSS 21 package program. Main results and the role of chance The median percentage (range) of the studied T cell subtypes was: T helpers (51.7%, 12.1%÷91.45%), T killers (39.0%, 4.1%÷72.5%), DPT cells (1.2%, 0÷8.75), DNT cells (8.4%, 1.9%÷46.2%), Treg cells (4.6%, 1.9%÷12.7%), TFH cells (7.6%, 1.4%÷42.6%). To examine the association between the percentage of Treg cells and the other T cell subtypes, the Spearman’s Rho Correlation coefficient for non-paramentric data was used. No correlation was found between the quantity of Treg and T helpers, T killers, Total T cells, double-positive T cells (p &amp;gt; 0.05). In contrast, the percentage of Treg showed a significant positive correlation with the percentage of TFH (R = 0.36; p = 0.01), and a significant negative correlation with double-negative T cells (R= - 0.29; p = 0.04). Limitations, reasons for caution The main limitation of the study is that it was carried out only in a limited period of time during the pregnancy. Thus, it would be beneficial to compare the percentage of Treg cells with other T cell sub-types in the second and third trimester of pregnancy. Wider implications of the findings Our study demonstrated that Treg cells have stronger association with TFH cells than with other T cell subtypes. The strength of this quantitative relationship might be important for the normal progress of pregnancy, and it remains subject of further studies. Trial registration number not applicable

Genomic instability and oxidative stress markers in pregnant women presenting fetal malformation

ObjectivesTo evaluated the oxidative stress and genomic instability in the peripheral blood of pregnant women with fetal malformation (PWFM), pregnant women (PW) and non-pregnant women (NPW). MethodsProtein carbonyl test (CP), thiobarbituric acid reactive substances test (TBARS), catalase enzyme activity (CAT), micronucleus assay with blocking cell cytokinesis - cytome (CBMN - cyt) and myeloperoxidase (MPO) assay were performed. Fetal karyotype and alpha-fetoprotein (AFP) in combination with ultrasound, were assessed to allow the diagnosis of fetal malformation. ResultsAn increase in CP, CAT and micronucleus was observed in the third trimester of pregnancy of PW compared to NPW. PWFM presented similar parameter analysis to PW, as well as in the comparison of the second trimesters of this groups. PWFM in the third gestational trimester showed a reduction in CP, nucleoplasmic bridge (NPB) and CAT compared to the same gestational period of PW. Considering the complete sample, participants who usually drank coffee daily had an increased frequency of nuclear buds (NBUD); CAT was decreased in whites compared to other ethnic groups. ConclusionsPregnancy is a physiological process that can lead to changes in oxidative stress markers and genomic instability through its course and compared to non-pregnant women.

Elevation of KIR+CD8+ T cells during human pregnancy

Abstract Recently, we identified KIR+CD8+ T cells as a previously underappreciated regulatory subset important in humans and mice to suppress self-reactivity, especially in the context of infections. To better understand what other roles these cells might play, we asked whether they are active during pregnancy. We first observed an increased frequency of KIR+CD8+ T cells in the peripheral blood of pregnant women at the second trimester compared to age-matched nonpregnant females. Women with a male fetus had a significantly higher level of KIR+CD8+ T cells than those with a female fetus. In vitro, KIR+CD8+ T cells suppressed HY-specific CD8+ T cells only from mothers with a male fetus. Therefore, the higher induction of KIR+CD8+ T cells in mothers carrying a male fetus may suppress the additional allogenic immune responses triggered by the Y chromosome from the male fetus. Longitudinal analysis over pregnancy showed that KIR+CD8+ T cells undergo expansion and differentiate into functional cytotoxic cells during pregnancy, and that their levels are maintained postnatally. In addition, increased levels of KIR+CD8+ T cells correlated with pregnancy disorders. Taken together, our findings suggest an important role of KIR+CD8+ T cells in the maintenance of maternal tolerance by suppressing fetal-specific alloreactive T cells. They may also be useful as predictive biomarkers or drug targets for human pregnancy disorders.

Phenotypic characterisation of regulatory T cells in patients with gestational diabetes mellitus

Gestational diabetes mellitus (GDM) is a common complication that occurs during pregnancy. Emerging evidence suggests that immune abnormalities play a pivotal role in the development of GDM. Specifically, regulatory T cells (Tregs) are considered a critical factor in controlling maternal–fetal immune tolerance. However, the specific characteristics and alterations of Tregs during the pathogenesis of GDM remain poorly elucidated. Therefore, this study aimed to investigate the changes in Tregs among pregnant women diagnosed with GDM compared to healthy pregnant women. A prospective study was conducted, enrolling 23 healthy pregnant women in the third trimester and 21 third-trimester women diagnosed with GDM. Participants were followed up until the postpartum period. The proportions of various Treg, including Tregs, mTregs, and nTregs, were detected in the peripheral blood of pregnant women from both groups. Additionally, the expression levels of PD-1, HLA-G, and HLA-DR on these Tregs were examined. The results revealed no significant differences in the proportions of Tregs, mTregs, and nTregs between the two groups during the third trimester and postpartum period. However, GDM patients exhibited significantly reduced levels of PD-1+ Tregs (P < 0.01) and HLA-G+ Tregs (P < 0.05) in the third trimester compared to healthy pregnant women in the third trimester. Furthermore, GDM patients demonstrated significantly lower levels of PD-1+ mTregs (P < 0.01) and HLA-G+ (P < 0.05) mTregs compared to healthy pregnant women in the third trimester. Overall, the proportion of Tregs did not exhibit significant changes during the third trimester in GDM patients compared to healthy pregnant women. Nevertheless, the observed dysregulation of immune regulation function in Tregs and mTregs may be associated with the development of GDM in pregnant women.

CD56 and Tim-3 molecule expression in different monocyte subsets in physiological pregnancy

Monocytes play an important role in the systemic immune defense against pathogens and maintaining physiological pregnancy. During pregnancy peripheral monocytes migrate into the decidua and form the pool of decidual macrophages which participate in the formation and development of placental tissues. The population of peripheral blood monocytes is phenotypically and functionally heterogeneous. In humans, there are different monocyte subsets depending on the expression of CD14 and CD16. CD56-positive monocytes are found in healthy women. Their number is positively correlated with body mass index, body fat. Tim-3 (T cell Ig and mucin domain-containing protein 3) expression is observed in peripheral monocytes during pregnancy. It is known that peripheral monocyte functions effectively change at pregnancy to form the immune tolerance at the maternal-fetal interface and the systemic immune defense against pathogens. However, the monocyte phenotype shift during pregnancy remain poorly understood. Therefore, the aim of the study was to evaluate the CD56 and Tim-3 expressions in monocyte subsets in human pregnancy. Peripheral blood mononuclear cells were isolated from peripheral blood of pregnant women (gestational age 29 weeks (28-31) by density gradient centrifugation and analyzed by flow cytometry. Peripheral blood of healthy non-pregnant fertile women (in follicular phase of the menstrual cycle) aged 21-29 years was studied as control. Pregnant women had a lower percentage of classical CD14hi/CD16- monocytes in comparison with non-pregnant. The percentages of intermediate (CD14hi/CD16+) and non-classical (CD14low/CD16+) monocytes did not change. The CD56 molecule expression was observed in all monocyte subsets in pregnant and non-pregnant women. Pregnant women had a higher percentage of CD56-positive classical (CD14hiCD16-) and non-classical (CD14lowCD16+) monocytes than non-pregnant. The percentage of CD56-positive intermediate (CD14hiCD16+) monocytes did not change. The percentages of double-positive CD56+Tim-3+ classical (CD14hiCD16-) and non-classical (CD14lowCD16+) monocytes were increased in pregnant women. The numbers of double-positive CD56+Tim-3+intermediate (CD14hiCD16+) monocytes did not change. Thus, the CD56 and Tim-3 expressions in different monocyte subsets were changed in human pregnancy.

The increased cfRNA of TNFSF4 in peripheral blood at late gestation and preterm labor: its implication as a noninvasive biomarker for premature delivery.

Given the important roles of immune tolerance and inflammation in both preterm and term labor, some inflammation-related genes could be related to the initiation of labor, even preterm labor. Inspection of cell-free RNA (cfRNA) engaged in inflammation in maternal blood may represent the varied gestational age and may have significant implications for the development of noninvasive diagnostics for preterm birth. To identify potential biomarkers of preterm birth, we investigated the cfRNA and exosomal miRNA in the peripheral blood of pregnant women at different gestational ages that undergo term labor or preterm labor. 17 inflammatory initiation-related cfRNAs were screened by overlapping with the targets of decreasing miRNAs during gestation and highly expressed cfRNAs at late gestation in maternal blood. To reveal the origins and mechanisms of these screened cfRNAs, the datasets of single-cell RNA sequencing from peripheral blood mononuclear cells of pregnant women, the fetal lung, and the placenta across different gestational ages were analyzed. During late gestation, TNFSF4 expression increased exclusively in pro-inflammatory macrophages of maternal blood, whereas its receptor, TNFRSF4, increased expression in T cells from the decidua, which suggested the potential cell-cell communication of maternally-originated pro-inflammatory macrophages with the decidual T cells and contributed to the initiation of labor. Additionally, the cfRNA of TNFSF4 was also increased in preterm labor compared to term labor in the validation cohorts. The EIF2AK2 and TLR4 transcripts were increased in pro-inflammatory macrophages from both fetal lung and placenta but not in those from maternal mononuclear cells at late gestation, suggesting these cfRNAs are possibly derived from fetal tissues exclusively. Moreover, EIF2AK2 and TLR4 transcripts were found highly expressed in the pro-inflammatory macrophages from decidua as well, which suggested these specific fetal-origin macrophages may function at the maternal-fetal interface to stimulate uterine contractions, which have been implicated as the trigger of parturition and preterm labor. Taken together, our findings not only revealed the potential of peripheral TNFSF4 as a novel cfRNA biomarker for noninvasive testing of preterm labor but further illustrated how maternal and fetal signals coordinately modulate the inflammatory process at the maternal-fetal interface, causing the initiation of term or preterm labor.

Epigenome wide association study in peripheral blood of pregnant women identifies potential metabolic pathways related to gestational diabetes

ABSTRACT Gestational diabetes mellitus (GDM) increases the risk of developing metabolic disorders in both pregnant women and their offspring. Factors such as nutrition or the intrauterine environment may play an important role, through epigenetic mechanisms, in the development of GDM. The aim of this work is to identify epigenetic marks involved in the mechanisms or pathways related to gestational diabetes. A total of 32 pregnant women were selected, 16 of them with GDM and 16 non-GDM. DNA methylation pattern was obtained from Illumina Methylation Epic BeadChip, from peripheral blood samples at the diagnostic visit (26–28 weeks). Differential methylated positions (DMPs) were extracted using ChAMP and limma package in R 2.9.10, with a threshold of FDR <0.05, deltabeta >|5|% and B >0. A total of 1.141 DMPs were found, and 714 were annotated in genes. A functional analysis was performed, and we found 23 genes significantly related to carbohydrate metabolism. Finally, a total of 27 DMPs were correlated with biochemical variables such as glucose levels at different points of oral glucose tolerance test, fasting glucose, cholesterol, HOMAIR and HbA1c, at different visits during pregnancy and postpartum. Our results show that there is a differentiated methylation pattern between GDM and non-GDM. Furthermore, the genes annotated to the DMPs could be implicated in the development of GDM as well as in alterations in related metabolic variables.

Regulation of fetal tolerance by KIR +regulatory CD8 +T cells in human pregnancy

Abstract Immune responses during pregnancy need to be precisely regulated to protect the fetus from microbial infections and to maintain tolerance for the semi-allogeneic fetus. However, the mechanisms regulating fetal tolerance during pregnancy are not well understood. Recently we have identified KIR +CD8 +T cells as a previously unappreciated subset with regulatory functions in humans to suppress harmful self-reactivity. Therefore, we asked whether KIR +CD8 +T cells also play a role in inducing immune tolerance during pregnancy. We first observed an increased frequency of KIR +CD8 +T cells, but not CD4 +regulatory T cells (Tregs), in the peripheral blood of pregnant women at second trimester compared to age-matched nonpregnant females. Interestingly, those with a male fetus had an even higher level of KIR +CD8 +T cells than the ones with a female fetus. In vitro, we found that KIR +CD8 +T cells can suppress CD8 +T cells specific to H-Y antigens (encoded by the Y chromosome) only in mothers with a male fetus. Therefore, the higher induction of KIR +CD8 +T cells in mothers carrying a male fetus may suppress the additional allogenic immune responses triggered by the Y chromosome from the male fetus. Moreover, KIR +CD8 +T cells expand, lose memory features and gain cytotoxic capacity along gestation and their levels are maintained postpartum. In addition, we found the numbers of these suppressive CD8 +T cells correlated with pregnancy disorders, such as spontaneous abortion or pre-eclampsia. Taken together, our findings suggest an important role of KIR +CD8 +T cells in the maintenance of fetal tolerance by suppressing alloreactive fetus-specific T cells, therefore they may be useful as predictive biomarkers or drug targets for human pregnancy disorders. Supported by Howard Hughes Medical Institute.

Study on the Correlation Between SFRP-5 Expression Level, Insulin Resistance, and Glycolipid Metabolism in Gestational Diabetes Mellitus

Objective: To investigate the correlation between serum secretory frizzled-related protein 5 (SFRP-5) expression levels and insulin resistance and glucolipid metabolism in patients with gestational diabetes mellitus (GDM). Methods: Baseline data were collected from 58 patients with GDM and 51 healthy controls who were admitted Affiliated Hospital of Hebei University from May 2020 to June 2022. sSTRA5 concentrations in peripheral blood of pregnant women were measured, and SFRP-5 levels in patients with different GDM types and normal controls were analyzed by logistic regression models. Results: The levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), fasting blood glucose (FBG), fasting insulin (FINS), hemoglobin A1c (HbA1c), and homeostasis model assessment-estimated insulin resistance (HOMA-IR) were higher in the observation group than in the control group, with statistically significant differences (P &lt; 0.05), while the expression levels of high-density lipoprotein cholesterol (HDL-C) and serum SFRP-5 were lower than in the control group, with statistically significant differences (P &lt; 0.05); serum SFRP-5, TG, TC, FBG, and HOMA-IR were all risk factors for GDM (P &lt; 0.05). Conclusion: Elevated serum sSTRA5 may be involved in the regulation of insulin resistance in the body and the regulation of blood glucose in the body by affecting lipid metabolism and inflammatory response.

Correlation between the intestinal microflora and peripheral blood Th1/Th2 balance in hypothyroidism during the first half of pregnancy.

This study aimed to investigate the relationship between intestinal microflora characteristics and the peripheral blood T helper cell (Th)1/Th2 balance in patients with hypothyroidism during the first half of pregnancy. The Th1/Th2 ratios in the peripheral blood of pregnant women in the hypothyroidism and control groups were determined using flow cytometry. The cytometric bead array assay was used to determine the serum levels of interleukin-2 (IL-2), IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ. Moreover, 16S rRNA amplicon sequencing was used to determine the intestinal microbial composition in the two groups. Finally, the relationships between intestinal microflora, Th1/Th2 cells, cytokines, and clinical indicators were analyzed. C-reactive protein levels were higher in the hypothyroidism group than in the control group. In contrast to the control group, the hypothyroidism group showed an increase in Th1 cells and the Th1/Th2 ratio, and a decrease in Th2 cells. The hypothyroidism group had higher serum IL-2, TNF-α, and IFN-γ levels, and lower IL-10 levels, than the control group. The richness of the intestinal microflora in the hypothyroidism group increased whereas the diversity decreased. The linear discriminant analysis effect size revealed that the hypothyroidism group had a higher abundance of Prevotella and Faecalibacterium, but a lower abundance of Bacteroides, compared to the control group. Prevotella was positively correlated with Th1 cells, the Th1/2 ratio, and TNF-α. Bacteroides was positively correlated with Th2 cells and IL-10, but negatively correlated with Th1 cells, the Th1/2 ratio, TNF-α, and IFN-γ. The thyroid peroxidase antibody level was directly proportional to TNF-α. A Th1/Th2 imbalance occurs in patients with hypothyroidism during the first half of pregnancy. Disorders of the intestinal microflora may lead to hypothyroidism during pregnancy by affecting the Th1/Th2 balance.

Maternal Serum tRNA-Derived Fragments (tRFs) as Potential Candidates for Diagnosis of Fetal Congenital Heart Disease.

Congenital heart disease (CHD) is one of the most predominant birth defects that causes infant death worldwide. The timely and successful surgical treatment of CHD on newborns after delivery requires accurate detection and reliable diagnosis during pregnancy. However, there are no biomarkers that can serve as an early diagnostic factor for CHD patients. tRNA-derived fragments (tRFs) have been reported to play an important role in the occurrence and progression of numerous diseases, but their roles in CHD remains unknown. High-throughput sequencing was performed on the peripheral blood of pregnant women with an abnormal fetal heart and a normal fetal heart, and 728 differentially expressed tRFs/tiRNAs were identified, among which the top 18 tRFs/tiRNAs were selected as predictive biomarkers of CHD. Then, a quantitative reverse transcriptase polymerase chain reaction verified the expression of tRFs/tiRNAs in more clinical samples, and the correlation between tRFs/tiRNAs abnormalities and CHD was analyzed. tRF-58:74-Gly-GCC-1 and tiRNA-1:35-Leu-CAG-1-M2 may be promising biomarkers. Through further bioinformatics analysis, we predicted that TRF-58:744-GLy-GCC-1 could induce CHD by influencing biological metabolic processes. Our results provide a theoretical basis for the abnormally expressed tRF-58:74-Gly-GCC-1 in maternal peripheral blood as a new potential biomarker for the accurate diagnosis of CHD during pregnancy.

Pyridoxal 5′-phosphate alleviates prenatal pyridaben exposure-induced anxiety-like behaviors in offspring

Pyridaben (PY) is a widely used organochlorine acaricide, which can be detected in the peripheral blood of pregnant women. Available evidence suggests that PY has reproductive toxicity. However, it remains uncertain whether prenatal PY exposure impacts neurobehavioral development in offspring. Here, we administered PY to pregnant mice at a dose of 0.5 and 5 mg kg−1 day−1 via gavage and observed anxiety-like behaviors in PY offspring aged five weeks. We then integrated the metabolome and transcriptome of the offspring's brain to explore the underlying mechanism. Metabolome data indicated that the vitamin B6 metabolism pathway was significantly affected, and the pyridoxal 5′-phosphate (PLP) concentration and the active form of vitamin B6 was significantly reduced. Moreover, the transcriptome data showed that both PLP generation-related Pdxk and anxiety-related Gad1 were significantly down-regulated. Meanwhile, there was a decreasing trend in the concentration of GABA in the hippocampal DG region. Next, we supplemented PLP at a dose of 20 mg kg−1 day−1 to the PY offspring via intraperitoneal injection at three weeks. We found up-regulated expression of Pdxk and Gad1 and restored anxiety-like behaviors. This study suggests that prenatal exposure to PY can disrupt vitamin B6 metabolism, reduce the concentration of PLP, down-regulate the expression levels of Pdxk and Gad1, inhibit the production of GABA, and ultimately lead to anxiety-like behaviors in offspring.

MicroRNA analysis in maternal blood of pregnancies with preterm premature rupture of membranes reveals a distinct expression profile.

To determine the expression profile of microRNAs in the peripheral blood of pregnant women with preterm premature rupture of membranes (PPROM) compared to that of healthy pregnant women. This was a pilot study with case-control design in pregnant patients enrolled between January 2017 and June 2019. Patients with healthy pregnancies and those affected by PPROM between 20- and 33+6 weeks of gestation were matched by gestational age and selected for inclusion to the study. Patients were excluded for multiple gestation and presence of a major obstetrical complication such as preeclampsia, diabetes, fetal growth restriction and stillbirth. A total of ten (n = 10) controls and ten (n = 10) patients with PPROM were enrolled in the study. Specimens were obtained before administration of betamethasone or intravenous antibiotics. MicroRNA expression was analyzed for 800 microRNAs in each sample using the NanoString nCounter Expression Assay. Differential expression was calculated after normalization and log2- transformation using the false discovery rate (FDR) method at an alpha level of 5%. Demographic characteristics were similar between the two groups. Of the 800 miRNAs analyzed, 116 were differentially expressed after normalization. However, only four reached FDR-adjusted statistical significance. Pregnancies affected by PPROM were characterized by upregulation of miR-199a-5p, miR-130a-3p and miR-26a-5p and downregulation of miR-513b-5p (FDR adjusted p-values <0.05). The differentially expressed microRNAs participate in pathways associated with altered collagen and matrix metalloprotease expression in the extracellular matrix. Patients with PPROM have a distinct peripheral blood microRNA profile compared to healthy pregnancies as measured by the NanoString Expression Assay.

Latest Research Findings on Prediction of Preeclampsia in Pregnant Women Based on Analysis of Cell-Free RNA in Peripheral Blood

Preeclampsia gravely threatens the health of mothers and infants. At present, treatment based on the relevant mechanisms of pathogenesis is still not available, and there is no independent reliable clinical index for early prediction of preeclampsia. According to recent studies, analysis of the cell-free RNA in the peripheral blood of pregnant women has shown that testing certain cell-free RNA levels can help predict in advance the occurrence of preeclampsia before clinical symptoms appear. In this paper, we described the status of research and progress in using maternal cell-free RNA analysis in predicting preeclampsia. In addition, we stated that cell-free RNA in peripheral blood may become a promising, real-time and non-invasive monitoring method that can be used to explore the mechanisms of pathogenesis and pathophysiology of preeclampsia and to identify different subtypes of preeclampsia.

Expression of inhibitory receptors PD-1, CTLA-4, and Tim-3 by peripheral T cells during pregnancy

Background. Inhibitory receptors and their ligands (also called checkpoint molecules) are important feedback regulators of the immune response. However, their role in immunological adaptation during pregnancy remains poorly understood.The aim of the study was to evaluate the level of checkpoint molecule (PD-1, CTLA-4, Tim-3) expression in peripheral T cells in pregnant women compared with fertile non-pregnant women.Materials and methods. The study included 36 women in the second half of pregnancy without pregnancy complications, 12 of whom had extragenital pathology. The control group consisted of 28 age-matched fertile non-pregnant women. The proportion of CD8+PD-1+, CD8+TIM-3+, CD8+PD-1+TIM-3+, CD4+PD-1+, CD4+TIM-3+, and CD4+PD-1+TIM-3+ was evaluated by flow cytometry using the corresponding monoclonal antibodies (BD Biosciences, USA).Results. The proportion of CD4+Tim-3+ and CD8+PD-1+ Т cells and CD4+ and CD8+ Т lymphocytes co-expressing PD-1 and Tim-3 in the peripheral blood of pregnant women was statistically significantly higher than in non-pregnant women. An increase in CD4+Tim-3+ and CD8+PD-1+ T cells was observed both in pregnant women with and without extragenital pathology. However, pregnant women with extragenital pathology were characterized by a higher CD8+PD-1+ count and a smaller number of CD8+Tim-3+ cells, as well as by a lack of an increase in PD-1+Tim-3+ T cells typical of pregnant women. The number of comorbidities was directly correlated with the proportion of CD8+PD-1+ lymphocytes and inversely correlated with the proportion of CD8+Tim-3+ and CD4+ PD-1+Tim-3+ cells. In addition, the expression of checkpoint molecules was associated with gestational age (a direct correlation was found with the proportion of CD8+Tim-3+, CD4+PD-1+Tim-3+, and CD8+PD-1+Tim-3+ cells) and to a lesser extent – with the age of pregnant women (an inverse relationship was found with the proportion of CD8+Tim-3+ cells).Conclusion. Pregnant women in the second half of pregnancy are characterized by increased expression of PD-1 and Tim-3 molecules in peripheral T cells. At the same time, concomitant extragenital pathology affects the expression of these molecules.

High-progesterone environment preserves T cell competency by evading glucocorticoid effects on immune regulation.

Progesterone (P4) and glucocorticoid (GC) play crucial roles in the immunoregulation of a mother to accept and maintain a semi-allogenic fetus. P4 concentration increases during pregnancy and becomes much higher in the placenta than in the other peripheral tissues, wherein the concentration of cortisol (COR), the most abundant GC and a strong immunosuppressor, remains uniform throughout the rest of the body. Here, we evaluated the effect of a high-P4 environment on pregnant immunity by comparing it with COR. Naïve T cell proportion increased transiently in peripheral blood of pregnant women just after delivery and decreased after one month. T cells stimulated with superantigen toxic-shock-syndrome-1 (TSST-1) in the presence of P4 stayed in the naïve state and did not increase, irrespective of the presence of COR, and reactive T cells could not survive. Treatment of T cells with P4 without T cell receptor (TCR) stimulation transiently suppressed T cell activation and proliferation, whereas the levels remain unaltered if P4 was not given before stimulation. Comparison of the engraftment and response against specific antigens using hu-PBL-NOG-hIL-4-Tg mice showed that P4-pretreated lymphocytes preserved CD62L expression and engrafted effectively in the spleen. Moreover, they produced antigen-specific antibodies, whereas COR-pretreated lymphocytes did not. These results suggest that a high-P4 environment suppresses T cell activation and induces T cell migration into lymphoid tissues, where they maintain the ability to produce anti-pathogen antibodies, whereas COR does not preserve T cell function. The mechanism may be pivotal in maintaining non-fetus-specific T cell function in pregnancy.

MiR-31-5p from placental and peripheral blood exosomes is a potential biomarker to diagnose preeclampsia

BackgroundPreeclampsia, a multisystem disorder of unknown etiology, is one of the leading causes of maternal and perinatal morbidity and mortality. Identifying sensitive, noninvasive markers can aid its prevention and improve prognosis. microRNAs (miRs), which function as negative regulators of gene expression, are closely related to preeclampsia occurrence and development. Herein we investigated the relationship between the DLK1-Dio3 imprinted miR cluster derived from placental and peripheral blood exosomes of pregnant women with preeclampsia and routine clinical diagnostic indicators, and also determined its potential as a noninvasive diagnostic marker.MethodsExosomes were extracted from the placenta and peripheral blood of pregnant women with preeclampsia.ResultsqPCR data indicated that the expression level of miRs, such as miR-134, miR-31-5p, miR-655, miR-412, miR-539, miR-409, and miR-496, in pregnant women with preeclampsia was significantly lower than that in healthy controls; miR-31-5p expression was the most different. Gene ontology analysis predicted that genes negatively regulated by miR-31-5p were mainly enriched in cellular entity, cellular process, and binding; moreover, Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that genes were involved in gonadotropin-releasing hormone receptor pathway and other signaling pathways. Correlation analysis revealed that miR-31-5p was significantly negatively correlated with clinical indicators of preeclampsia, such as systolic and diastolic pressure, lactate dehydrogenase, and proteinuria.ConclusionWe believe that exosome-derived miR-31-5p can serve as an effective and sensitive biomarker to determine the course of preeclampsia in pregnant women.

Noninvasive prenatal prediction of fetal haplotype with Spearman rank correlation analysis model.

BackgroundNoninvasive prenatal testing (NIPT) has been widely used clinically to detect fetal chromosomal aneuploidy with high accuracy rates, gradually replacing traditional serological screening. However, the application of NIPT for monogenic diseases is still in an immature stage of exploration. The detection of mutations in peripheral blood of pregnant women requires precise qualitative and quantitative techniques, which limits its application. The bioinformatic strategies based on the SNP (single nucleotide polymorphism) linkage analysis are more practical, which can be divided into two types depending on whether proband information is needed. Hidden Markov Mode (HMM) and Sequential probability ratio test (SPRT) are suitable for families with probands. In contrast, methods based on databases and population demographic information are suitable for families without probands.MethodsIn this study, we proposed a Spearman rank correlation analysis method to infer the fetal haplotypes based on core family information. Allele frequencies of SNPs that were used to construct parental haplotypes were calculated as sets of nonparametric variables, in contrast to their theoretical values represented by a fetal fraction (FF). The effects on the calculation of the fetal concentration of two DNA enrichment methods, multiple‐PCR amplification, and targeted hybrid capture, were compared, and the heterozygosity distribution of SNPs within pedigrees was analyzed to reveal the best conditions for the model application.ResultsPredictions of the paternal haplotype inheritance were in line with expectations for both DNA library construction methods, while for maternal haplotype inheritance prediction, the rates were 96.55% for method multiple‐PCR amplification and 95.8% for method targeted hybrid capture.ConclusionPositive prediction rates showed that the maternal haplotype prediction was not as accurate as paternal one, due to the large amount of maternal noise in the mother's peripheral blood. Although this model is relatively immature, it provides a new perspective for noninvasive prenatal clinical tests of monogenic diseases.

The correlation between proteoglycan 2 and neuropsychiatric systemic lupus erythematosus

The proposed pathogenesis of neuropsychiatric systemic lupus erythematosus (NPSLE) mainly includes ischemia and neuroinflammation mechanisms. Protein encoded by Proteoglycan 2 (PRG2) mRNA is involved in the immune process related to eosinophils, also being found in the placenta and peripheral blood of pregnant women. We evaluated the correlation between PRG2 and NPSLE for the first time and found that PRG2 protein was overexpressed in the serum of patients with NPSLE and correlated with the SLE disease activity index (SLEDAI) subset scores of psychosis. Moreover, we investigated the correlation between hippocampal PRG2 level and hippocampally dependent learning and memory ability in MRL/lpr mice, and discovered that the number of PRG2+GFAP+ astrocytes in the cortex and hypothalamus and the number of PRG2+IBA-1+ microglia in the hippocampus and cortex significantly increased in the MRL/lpr mice. These data provided a reference for the follow-up exploration of the role of PRG2 in SLE or other diseases.