Distribution of mononuclear cells in peripheral blood of females with spontaneous abortion]

The quantitative distribution of peripheral T cells, monocytes/granulocytes, natural killer cells and B cells was estimated in aborting women using the peroxidase-antiperoxidase technique with monoclonal antibodies. 30 women with spontaneous abortions were included in the study and 30 fertile and 50 healthy pregnant women as control groups. It could established that the normal pregnancy has no influence on the distribution of mononuclear cells. In cases with threatened abortions was the number of monocytes/granulocytes and of natural killer cells increased if the loss of pregnancy occurred. The differentiation of mononuclear cells may be helpful for the prognosis of abortions.     

Increased inducibility of inflammatory mediators from peripheral blood mononuclear cells of women with salpingitis

To investigate whether immune system activation may contribute to the tissue damage observed in salpingitis, we isolated peripheral blood mononuclear cells and quantitated production of the monocyte activation products tumor necrosis factor-alpha, interleukin-1, and interleukin-6. Unstimulated cells from 7 of 20 women with salpingitis spontaneously released tumor necrosis factor at a concentration greater than 2 SD above the mean value produced by cells from 29 healthy donors. Interferon gamma (200 U/ml) further induced production of tumor necrosis factor from mononuclear cells of 11 women with salpingitis. In contrast, production of tumor necrosis factor by each of 23 other patients who lacked laparoscopic or clinical evidence of salpingitis was similar to that of the controls. In a subset of women whose cells were tested for production of other monokines, three of nine women with salpingitis spontaneously released interleukin-1 but none of the others did so. Four of nine patients with salpingitis also produced interleukin-6, but none of the others did so. None of the monokines were detected in serum from any subject. The results suggest that monocytes from women with salpingitis are primed in vivo and produce inflammatory mediators under conditions where monocytes from other women are poorly responsive. This increased monokine inducibility may contribute to the tubal damage that is the hallmark of salpingitis.     

Activated peripheral blood mononuclear cells induce p44/42 mitogen-activated protein kinase phosphorylation in trophoblast-like JAR cells

Mammalian pregnancy bears many similarities to transplantation, since the fetus is semi-allogenic to mother. Thus, mammals have developed numerous mechanisms to protect the developing fetus from maternal immunologic recognition and attack. We have previously shown that human choriocarcinoma JAR cells, which resemble first trimester trophoblasts, regulate several important mRNAs in activated peripheral blood mononuclear cells (PBMC). We now provide further evidence that communication between maternal and fetal tissues is bi-directional, and that activation of PBMC leads to activation of specific signaling pathways in JAR cells. Activated PBMC were co-cultured with JAR cells for specific time intervals, after which JAR cells were lysed and subjected to western blotting for activated forms of the JNK, Erk 1-2, and p38 mitogen-activated protein kinases (MAPK). Phosphorylation of Erk 1-2, but not JNK or p38, was induced in co-cultures of PBMC and JAR cells. These results were also obtained when JAR cells were incubated with conditioned medium from activated, but not resting, PBMC. Results were confirmed using specific MAPK reporter constructs, using luciferase activity as a measure of Elk-1 phosphorylation. Erk 1-2 phosphorylation was not required for JAR cells to inhibit IL-2 production in activated PBMC. Addition of the specific MAPK inhibitor UO126 to JAR cells prior to the addition of activated PBMC to the cultures did not abolish the capacity of JAR cells to inhibit IL-2 mRNA expression in PBMC. We conclude that there is likely to be significant bi-directional signaling between leukocytes and trophoblasts at the maternal–fetal interface. We propose the existence of a delicate maternal–fetal immunologic homeostasis based on these experimental results.     

B族链球菌携带及感染孕妇外周血单核细胞炎症因子的水平变化#br#

目的分离培养B族链球菌(GBS)感染孕妇外周血单核细胞,探究GBS感染后炎症因子的变化。方法选择2017年6月至2018年12月在青岛市海慈医疗集团产科分娩的GBS阳性孕妇和正常孕妇,以及育龄期妇女作为研究对象,分离培养受试者外周血单核细胞,探究IL-1、IL-6、IL-8、TNF-α、CRP的m RNA及蛋白表达量的变化。结果GBS感染组外周血单核细胞IL-6(102.22±98.62)ng/L和IL-8(98.17±87.12)ng/L的水平显著高于正常孕妇组IL-6(24.62±53.14)ng/L、IL-8(20.25±30.11)ng/L和育龄期妇女组IL-6(22.13±56.23)ng/L、IL-8(17.12±20.63)ng/L(P<0.001)。GBS感染组新生儿发病率为20.0%,显著高于正常孕妇组(0.0%;P<0.05)。结论GBS感染阳性孕妇外周血单个核细胞中,IL-6和IL-8炎症因子表达显著上升,且新生儿发病风险增加,临床上应给予重视。     

Expression levels of seven candidate genes in human peripheral blood mononuclear cells and their association with preeclampsia

Objective: To evaluate the peripheral blood mononuclear cell (PBMC) expression levels of hemeoxygenase 1 (HMOX-1), superoxide dismutase 1 (SOD-1), vascular endothelial growth factor A (VEGF-A), transforming growth factor beta 1 (TGF-β1), interleukin (IL)-6, IL-15 and AdipoQ genes to study their association with preeclampsia (PE). Methods: A total of 177 pregnant women were recruited: 108 cases and 69 controls. Quantification of gene expression was measured by quantitative real-time polymerase chain reaction (PCR) using TaqMan probes. Results: Underexpression of VEGF-A and TGF-β1 was a constant in most of the cases (80.91% and 76.36%, respectively) and their expression was associated with onset and/or severity of disease (p values?Conclusion: PBMC underexpression of VEGF-A and TGF-β1 is a hallmark of PE in the study population.     

Maternal blood coagulation factor XIII is associated with the development of cytotrophoblastic shell

We analysed the early implantation tissues of normal women and of a patient with congenital factor XIII deficiency in order to study the role of maternal subunit A of factor XIII (XIIIA) in the development of extravillous cytotrophoblast. The patient had received adequate administration of factor XIIIA concentrate only up to 7 weeks of gestation (wG). Her pregnancy was maintained until the latter half of 8 wG, but was terminated by intrauterine fetal death at 9 wG. Immunohistochemical staining of cytokeratin, XIIIA and subunit S of factor XIII was performed in the early implantation tissues of normal women and of this patient. Numerous well-formed cytotrophoblastic shells and Nitabuch's layers were detected in implantation tissues at 7-8 wG in normal women, and XIIIA was present in the intercellular space in well-formed cytotrophoblastic shells, while the cytotrophoblastic shells and Nitabuch's layers in this patient's implantation tissue were poorly-formed. Furthermore, XIIIA was not detected around them. It is suggested that when the maternal plasma activity of factor XIII is low, the concentration of XIIIA at the placental bed is also low, leading to the insufficient formation of cytotrophoblastic shell and therefore an increased probability of miscarriage in patients with congenital factor XIII deficiency.     

Type II estrogen binding sites in human peripheral blood mononuclear cells: variations during the menstrual cycle

We have previously reported that human peripheral blood mononuclear cells (PBMC) contain type II estrogen binding sites (type II EBS). In this study, the fluctuations of type II EBS during the menstrual cycle were analyzed in 6 normally menstruating women. Approximately 3 times higher levels of type II EBS were found in the periovulatory period with respect to both follicular and luteal phases. In postmenopausal women the mean type II EBS levels were similar to those observed in the follicular phase of the cycle. However, in 3 postmenopausal patients a short course of estrogen or tamoxifen resulted in a marked increase of type II EBS levels. Tamoxifen was also found to compete with 17 beta-estradiol for type II EBS in PBMC, although to a lesser extent than diethylstilbestrol.     

In-situ detection of both inflammatory and anti-inflammatory cytokines in resting peripheral blood mononuclear cells during pregnancy

INTRODUCTION: Local and possibly systemic curtailment of the maternal immune response is important for a successful pregnancy. Although the local milieu at the utero-placental interface is likely to harbor the most prominent alterations, it is suggested, at least in mice, that systemic immunity is also tolerized during pregnancy. In the present study, we investigated mRNA expression of the key immunomodulatory cytokines; interleukin (IL)-4, IL-10, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma during normal pregnancy. MATERIAL AND METHODS: In-situ hybridization (ISH) of cytokine mRNA in resting peripheral blood mononuclear cells (PBMCs) was used to detect the number of cells spontaneously expressing cytokines. Eleven women with normal gestations were followed during pregnancy as well as 8 weeks postpartum, and compared with 10 non-pregnant healthy controls. RESULTS: The numbers of IFN-gamma and IL-4 mRNA expressing cells were found to be significantly increased during pregnancy and postpartum compared with non-pregnant controls. Pregnant women and non-pregnant controls did not differ in their expression of TNF-alpha and IL-10. CONCLUSION: Our studies demonstrated increased numbers of both IFN-gamma and IL-4 mRNA expressing cells in blood suggesting that systemic immunomodulation, albeit partial, takes place during normal pregnancy. It is proposed that enhanced IL-4 expression, possibly in concert with other elevated anti-inflammatory immunomodulatory cytokines, curtail the potentially hazardous effects of IFN-gamma on systemic immunity during pregnancy.     

Intrauterine administration of autologous peripheral blood mononuclear cells increases clinical pregnancy rates in frozen/thawed embryo transfer cycles of patients with repeated implantation failure

Intrauterine administration of autologous peripheral blood mononuclear cells (PBMC) activated by HCG in vitro are reported to improve implantation rates in patients with repeated failure of IVF-ET. In this study, we examined the effects of intrauterine administration of freshly isolated PBMC on clinical pregnancy and the implantation rates of patients who received frozen/thawed embryo transfer by prospective cohort study. Patients who had not achieved a successful pregnancy despite at least one or more IVF-ET sessions were enrolled in this study (n = 253, 253 cycles). Based on the patient's treatment preferences, PBMC were freshly isolated from each patient and then administered to the intrauterine cavity of that patient. Frozen/thawed embryo transfer was performed and the success of implantation in the PBMC-treated group (n = 83, 83 cycles) was compared with that in the non-treated control groups (n = 170, 170 cycles). There were no significant differences in the clinical pregnancy rate (34.9% vs. 32.9%), implantation rate (21.6% vs. 21.1%) and live birth delivery rate (21.7% vs. 21.8%) between PBMC-treated and non-treated groups. However, when the analyses were restricted to patients who had three or more implantation failures, the clinical pregnancy rate and the implantation rate in the PBMC-treated group (42.1% and 25.0%, p<0.05; n = 19 and 32, respectively) were significantly higher than those in the non-treated group (16.7% and 9.4%, p<0.05; n = 36 and 64, respectively). These findings indicate that intrauterine administration of autologous PBMC freshly isolated from patients, effectively improves embryo implantation in patients with three or more IVF failures.     

Preeclampsia is associated with a serum factor cytotoxic to human endothelial cells

Preeclampsia occurs in 7% to 10% of pregnancies and is a leading cause of morbidity for mothers and their infants. Intensive investigation has failed to reveal the cause of the multiple organ dysfunction characteristic of this disorder, which abates completely with delivery. However, several observations suggest that endothelial cell dysfunction is a central pathophysiologic event. We report that serum from preeclamptic women is cytotoxic to endothelial cells in vitro. Consistent with the reversal of the clinical condition after delivery, cytotoxic activity in serum of preeclamptic women is reduced after 24 to 48 hours post partum. In contrast, cytotoxic activity of serum from normal pregnant women increases after delivery.     

Intrauterine administration of autologous peripheral blood mononuclear cells increases clinical pregnancy rates in frozen/thawed embryo transfer cycles of patients with repeated implantation failure

Intrauterine administration of autologous peripheral blood mononuclear cells (PBMC) activated by HCG in vitro are reported to improve implantation rates in patients with repeated failure of IVF-ET. In this study, we examined the effects of intrauterine administration of freshly isolated PBMC on clinical pregnancy and the implantation rates of patients who received frozen/thawed embryo transfer by prospective cohort study. Patients who had not achieved a successful pregnancy despite at least one or more IVF-ET sessions were enrolled in this study (n = 253, 253 cycles). Based on the patient's treatment preferences, PBMC were freshly isolated from each patient and then administered to the intrauterine cavity of that patient. Frozen/thawed embryo transfer was performed and the success of implantation in the PBMC-treated group (n = 83, 83 cycles) was compared with that in the non-treated control groups (n = 170, 170 cycles). There were no significant differences in the clinical pregnancy rate (34.9% vs. 32.9%), implantation rate (21.6% vs. 21.1%) and live birth delivery rate (21.7% vs. 21.8%) between PBMC-treated and non-treated groups. However, when the analyses were restricted to patients who had three or more implantation failures, the clinical pregnancy rate and the implantation rate in the PBMC-treated group (42.1% and 25.0%, p < 0.05; n = 19 and 32, respectively) were significantly higher than those in the non-treated group (16.7% and 9.4%, p < 0.05; n = 36 and 64, respectively). These findings indicate that intrauterine administration of autologous PBMC freshly isolated from patients, effectively improves embryo implantation in patients with three or more IVF failures.     

儿童肺炎支原体肺炎金属基质蛋白酶9血清水平测定及其外周血单核细胞内的基因表达

目的探讨肺炎支原体肺炎(MPP)患儿的金属基质蛋白酶9(MMP-9)血清水平及其在外周血单核细胞内的基因表达,为MMP在免疫病理机制中的作用提供依据。方法选取2015年1月至2016年8月在深圳市宝安区妇幼保健院小儿呼吸科接受治疗和随访的急性MPP患儿78例作为研究对象,为观察组。同期选取在本院进行健康体检的儿童78例作为对照组。采用酶联免疫吸附法及实时定量PCR法分别检测血清MMP-9水平以及MMP-9在外周血单核细胞内的基因表达。结果观察组急性期患儿血清MMP-9水平显著高于恢复期,同时也高于对照组,差异有统计学意义(P0.05);观察组恢复期儿童血清MMP-9水平与对照组比较差异无统计学意义(P0.05)。观察组急性期患儿的外周血MMP-9mRNA阳性表达率高于恢复期,同时也高于对照组,差异有统计学意义(P0.05)。结论血清MMP-9和MMP-9mRNA水平测定对于判断病情具有重要意义。     

宫腔灌注自体外周血单个核细胞治疗对反复种植失败患者妊娠结局的影响及相关机制

目的探讨宫腔灌注自体外周血单个核细胞(PBMCs)治疗对反复种植失败(RIF)患者妊娠结局的影响及相关机制。方法 36名RIF患者于冻融胚胎移植(FET)前24 h行自体PBMCs宫腔灌注,随访妊娠结局;酶联免疫吸附试验(ELISA)检测宫腔灌注前、后外周血及宫颈分泌物中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平。结果 RIF患者经宫腔灌注自体PBMCs治疗后得到了较高的临床妊娠率(47.22%);宫颈分泌物中IL-6、IL-6/TNF-α均显著升高(P0.05),TNF-α水平无统计学差异(P0.05),外周血中IL-6、TNF-α水平均无统计学差异(P0.05)。结论宫腔灌注自体PBMCs治疗能提高RIF患者宫腔局部IL-6水平及IL-6/TNF-α比值,有效改善RIF患者临床妊娠率。     

Proliferation, interleukin-2 production and receptor expression in peripheral blood mononuclear cells from colorectal cancer patients

Peripheral blood mononuclear cells (PBMC) were cultured with phytohemagglutinin (PHA) in 46 colorectal cancer patients and 27 normal controls. By collectively comparing the cancer patients with the normal controls, the cancer patients had: (1) decreased interleukin-2 (IL-2) secretion, (2) fewer interleukin-2 receptor (IL-2R) expression on cell surface, and (3) less 3H thymidine incorporation. The ability of the phorbol ester, phorbol 12-myristate 13-acetate (PMA), and the calcium ionophore, A23187, in co-stimulation with PHA, to enhance the IL-2 secretion, IL-2R expression and 3H thymidine incorporation were studied in the PBMC of colorectal cancer patients. By adding A23187 into the PHA-stimulated PBMC from colorectal cancer patients, IL-2 production and IL-2R expression were increased up to levels equal to that obtained in PHA-stimulated PBMC from normal controls. Whereas there was no significant increase in proliferative response. However, PMA has no effect on all three parameters. A23187 is known to be effective in elevating cytosolic free Ca2+ and PMA is regarded as an activator of protein kinase C. Therefore, we may conclude that the impairment of IL-2 production and IL-2R expression in PHA-stimulated cultures is mainly due to failure of the free Ca2+ release which can be repaired by A23187. While protein kinase C activation may not be impaired since its deficit in IL-2 production and IL-2R expression can not be corrected by PMA.