新生儿IL-18对Th1/Th2类细胞免疫平衡的作用与HBV宫内感染相关性研究

目的:探讨新生儿IL-18对Th1/Th2类细胞免疫平衡的作用与HBV宫内感染的相关性。方法:选择HBsAg、HBeAg双阳性但肝功能正常孕产妇分娩的新生儿157例,以其中胎儿宫内感染21例为研究组,无宫内感染136例为对照组。用流式细胞仪检测两组脐血白细胞介素-18(IL-18)、γ-干扰素(IFN-γ)、白细胞介素-4(IL-4)的表达水平。结果:(1)研究组IL-18及IFN-γ表达水平明显低于对照组,差异有显著性(P<0.05;P<0.05);研究组IL-4表达水平百分数高于对照组,但差异无显著性(P>0.05);研究组IFN-γ/IL-4高于对照组,差异有显著性(P<0.05);(2)研究组IL-18与IFN-γ、IL-4、IFN-γ/IL-4均呈显著正相关(P均<0.05)。结论:IL-18是Th1/Th2网络平衡的更高层次的调控者;IL-18水平下调,导致新生儿Th1/Th2细胞因子比例下降,以致不利于HBV的清除,IL-18水平下调可能是HBV宫内感染的危险因素之一。     

Feto-maternal interface in T helper 1-type and T helper 2-type immunity in labor

Aim The aim of this study was to examine the relationship between Th (T helper) cells in maternal and umbilical cord blood in normal delivery.Methods The percentage of Th1 and Th2 cells and the Th1:Th2 cell ratios in maternal peripheral and umbilical venous blood from 61 normal singleton pregnancies at delivery at 37–41 weeks' gestation were calculated using flow cytometry.Results The proportion of Th cells and the Th1:Th2 ratio in the maternal blood were not correlated with those in the cord blood.Conclusion Our results indicate that the proportion of Th cells in cord blood does not depend on that of maternal Th cells at labor.     

The immunologic profile of infants born after maternal immunoglobulin treatment and intrauterine platelet transfusions for fetal/neonatal alloimmune thrombocytopenia

OBJECTIVE: The purpose of this study was to evaluate whether maternally administered intravenous immunoglobulins (IVIG) and intrauterine platelet transfusions (IUPT) for fetal/neonatal alloimmune thrombocytopenia (FNAIT) affect the development of the fetal immune system. STUDY DESIGN: The lymphocyte subset distribution of mononuclear cells of cord blood of 20 FNAIT newborns was analyzed by flow cytometry and compared with a control group of healthy newborns and a reference group treated with intrauterine erythrocyte transfusions (IUET) for hemolytic disease. RESULTS: The percentage of monocytes, natural killer (NK) cells, ratios of mature and immature T cells and B cells, and resting or activated cells were not significantly different compared to the control group. In addition, the B-cell and T-cell populations showed a normal in vitro antibody production and T-cell proliferation when compared with the control group. CONCLUSION: Antenatal treatment for FNAIT with maternal IVIG with or without IUPT is not associated with lymphocyte activation or premature maturation of the neonatal immune system.     

T helper 1- and T helper 2-type cytokine imbalance in pregnant women with pre-eclampsia.

OBJECTIVES: The purpose of our study was to investigate T helper 1/T helper 2 balance in pregnant women with pre-eclampsia. STUDY DESIGN: 18 patients with pre-eclampsia and 20 healthy pregnant women were included in the study. Peripheral blood mononuclear cells (PBMC) were stimulated with phytohaemagglutinin (PHA) for 48 h. Cytokine: interleukin-2 (11-2), interferon-gamma (IFN-gamma) and interleukin-10 (I1-10) concentrations in culture supernatants were determined using the ELISA method. Statistical analysis was performed using a standard non-parametric Mann-Whitney U-test. RESULTS: We found that in pre-eclamptic patients PHA-stimulated 11-2 and IFN-y production was significantly higher (P<0.001) and I1-10 production significantly lower (P<0.005) in comparison with the control group. CONCLUSION: These results could suggest that there is Th1/Th2 imbalance in pre-eclamptic patients with predominant Th1-type immunity.     

复发性流产患者外周血T淋巴细胞亚群及CD4^＋辅助性T淋巴细胞亚型的变化

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Nrf_2与足月胎膜早破合并绒毛膜羊膜炎的关系

目的:探讨核因子E2相关因子(Nrf_2)表达与足月胎膜早破(PROM)合并绒毛膜羊膜炎(HCA)的关系。方法:选取足月PROM患者60例,根据胎膜组织病理检查结果分成PROM合并HCA组(HCA组,21例)和未合并HCA组(对照组,39例)。采用免疫组化法、蛋白印迹法和实时荧光定量PCR法分别检测Nrf_2在胎膜组织、母血和脐血中的表达。结果:(1)HCA组胎膜组织、母血和脐血中Nrf_2mRNA表达水平分别为0.48±0.09、0.73±0.11和0.51±0.09,Nrf_2蛋白表达水平分别为0.101±0.019、0.294±0.051和0.175±0.028;HCA组中Nrf_2mRNA和蛋白表达水平均明显低于对照组,差异有统计学意义(P0.05)。(2)Nrf_2在胎膜组织羊膜层、绒毛膜层和底蜕膜层中的细胞核和细胞质均有表达。Nrf_2定位于细胞核的表达在HCA组的每一层表达均低于对照组(P0.05);两组中,Nrf_2在羊膜层、绒毛膜层和底蜕膜层中的表达水平比较,均存在显著差异,底蜕膜最低,羊膜层最高。结论:Nrf_2表达水平显著下降是PROM合并HCA的重要发病机制。Nrf_2含量变化可成为一种新的生物学标记物用于诊断PROM合并HCA的孕妇和评估新生儿预后。     

Distribution of Th1 and Th2 cell populations in human peripheral and decidual T cells from normal and anembryonic pregnancies

OBJECTIVE: To examine whether maternal immune responses during normal pregnancy are Th2 biased and whether there are specific changes when anembryonic pregnancy occurs. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology at a university hospital. PATIENT(S): We studied 32 pregnant women receiving elective abortions of normal pregnancies and 35 women with anembryonic pregnancies between 6 weeks and 10 weeks of gestational age. INTERVENTION(S): Using the multilabeling capability of three-color flow cytometry, it is possible to measure intracellular cytokines and cell surface markers simultaneously to determine which cells are the cytokine-producing cells. MAIN OUTCOME MEASURE(S): We examined the extent and proportion of mononuclear cells expressing specific T-cell surface markers and cytokines, interferon gamma, and interleukin 4 in the peripheral blood and deciduae. Secreted cytokines in the supernatants after 24-hour culture were also compared. RESULT(S): During the unstimulated status, the proportion of IL-4-secreting cells significantly exceeded that of IFN-gamma-secreting cells in the peripheral blood and decidua in normal pregnancies and was significantly decreased when anembryonic pregnancies occurred. Consequently, the Th1/Th2 ratios were increased during anembryonic pregnancies. However, after 24-hour culture, only another Th2-type cytokine, IL-10, was markedly increased and exceeded IFN-gamma secretion in cultures from both the peripheral blood and decidua in normal pregnancies. CONCLUSION(S): The decidual T lymphocytes are Th2 predominant. When anembryonic pregnancy occurs, this Th2 predominance disappears.     

Is the mother's bacterial vaginosis with PROM a significant factor for intrauterine infection of the fetus in preterm labor before 32 weeks of gestation

OBJECTIVE: Bacterial vaginosis of pregnant woman is an important factor of appearance of the PROM in result of intrauterine infections on every stage of pregnancy. Frequency exist of intrauterine infection evaluate from 1 to 10% pregnancies. Clinical chorio-amnionitis complicates 1-5% of term pregnancies, but nearly 25% of preterm deliveries and really increase morbidity and mortality of newborns and they are also a reason of mothers morbidity. A condition of recognition the intrauterine infections is ascertainment of infection factor risk of mother and appearance of clinic symptoms of newborns. The aim of this work was a valuation of dependence between bacterial flora taken from cervics uteri of pregnant woman with PROM who gave birth with preterm below 32 weeks and a presence of clinic and laboratory symptoms of infections and the presence of bacterial flora in blood culture of the newborns. MATERIAL AND METHODS: 37 pregnant women at the age from 20 to 43 hospitalized in Department Perinatology ICZMP between 1999-2001 because of PROM and a threat of preterm labor at the fetal age from 24 to 32 weeks were taken by an analysis. The average time of amniotic liquid outflowing before the labor was 11.1 days +/- 8.63, average length persistence of pregnancy was 27.6 hbd +/- 1.85, average born mass was 1029 g +/- 187.5. Laboratory factors and clinics symptoms chorionamnionitis, bacteriology cultures of swabs from cervics uteri and information of the labor of pregnant women were analyzed. Diagnostics of newborns contained: making of blood cultures and skin, nose, throat, urine cultures, estimation WBC and level CRP, estimation of lungs X-ray. RESULTS: In result of bacteriology cultures from cervics uteri of mothers gained bacteria of frequent occurrence: Staphylococcus epiderm.--27%, E. coli--19%, Streptococcus gr. B--10.8%, Enterococcus foecalis--8.1%. In result of bacteriology cultures of newborns blood gained bacteria of frequent occurrence: Staphylococcus epiderm.--13.5%, Staphylococcus haemolitycus--10.8%, Klebsiella pneumoniae--10.8%, Pseudomonas aeruginosa--8.1%. In 97.3% cases at newborns appeared inborn infections in a shape of pneumonia confirmed by X-ray. Confirmation of intrauterine infection in histopathology examination of the placenta in a shape of chorionamnionitis was shown in 25 cases, in other words in 67.5%. Positive results of pathogenic bacteriology flora cultures taken from cervics uteri was in 50% pregnant woman, however in 43% of newborns bacteriology cultures of the blood were positive. In none of the analyzed preterm labor cases with PROM bacterial flora from positive cultures taken from cervics uteri of mothers did not correlate with bacterial flora found in blood cultures of newborns. CONCLUSIONS: On a base of analyzed values PROM of mothers with bacterial vaginosis is the important factor risk of the evolution of intrauterine infections of newborns, besides did not ascertain the dependence between the values of bacteriology cultures taken from cervics uteri of mothers and the values of bacteriology cultures of the newborns blood with clinic symptoms of intrauterine infection.     

Synthesis of IFN-gamma by CD8(+) T cells is preserved in HIV-infected women with HPV-related cervical squamous intraepithelial lesions

OBJECTIVE: The aim of this study was to investigate whether coinfection with HIV affects the synthesis of Th1 and Th2 cytokines by peripheral blood T cells of women infected with human papillomavirus (HPV). METHODS: Cervical swabs and peripheral blood were obtained from women referred for colposcopy. HPV DNA by Digene's hybrid capture assay, HIV RNA by Roche's Amplicor assay, and cytokine synthesis of T-cell subsets by flow cytometry were assessed. HPV-associated cervical and HIV-associated immune deficiency diseases were staged using the Bethesda System and the Centers for Disease Control criteria, respectively. RESULTS: Patients with HIV and/or HPV infections had lower percentages of IL-2(+) and higher percentages of IL-10(+) T cells than healthy women. Furthermore, women with both virus infections (HIV(+)/HPV(+)) had significantly fewer IL-2(+) CD4(+), IFN-gamma(+) CD4(+), and TNF-alpha(+) CD4(+) T cells than women with HPV infection alone (HPV(+)). Whereas HIV(+) and healthy women had similar numbers of IFN-gamma(+) CD8(+) T cells, HPV(+) women had significantly fewer IFN-gamma(+) CD8(+) T cells than healthy women. CONCLUSION: HIV infection adversely affects the synthesis of Th1 cytokines by CD4(+), but not IFN-gamma synthesis by CD8(+) T cells of women with active HPV infection. The increase in IFNgamma(+) CD8(+) T cells, a phenotype consistent with cytotoxic T lymphocytes, may account for the stable HIV disease of the women studied. However, the increase in IFN-gamma(+) CD8(+) T cells is less likely to be HPV-specific as there was a higher incidence of HPV-related cervical SIL in HIV(+)/HPV(+) women compared with HPV(+) women.     

Immunoregulatory effects of human cord blood T lymphocytes on mixed lymphocyte reaction

Fetuses of outbred mammalian species can survive despite disparities of participants' histocompatibility antigens. During pregnancy, maternal immune response is supposed to be suppressed by various immune suppressive mechanisms. In order to know whether fetal lymphocytes can suppress the maternal immune reaction, we investigated the suppressive effect of human newborns' T lymphocytes. Newborn T cells were isolated from the umbilical cord blood by means of the sheep red blood cell rosette forming method and treated with mitomycin C. The newborn T cells were added as regulator cells to one way mixed lymphocyte reaction (MLR) using maternal lymphocytes, paternal lymphocytes and unrelated donor's lymphocytes. In a control study, adult T cells were treated in the same manner. In any combination of the MLR, the response was substantially suppressed by the addition of cord T cells in comparison with the response by the addition of normal adult T cells. The suppression was dose-related with an increasing number of regulator cord T cells. The suppressive activity of cord T cells showed radiation resistance until 1,000 rads irradiation. In the kinetics study, cord T cells were found to suppress the recognition phase of MLR. These results suggest that cord T lymphocytes suppress the mother's immunological reaction to the fetal antigens.     

Relation between plasma endothelin 1 levels and T helper 1: T helper 2 cell immunity in women with preeclampsia.

The aim of this study was to investigate the relationship between plasma endothelin 1 (ET-1) levels and T helper (Th)-1:Th2 cell immunity in women with preeclampsia. The percentage of Th1 and Th2 cells and the Th1:Th2 cell ratios in peripheral blood from 11 normal pregnant women and 11 patients with preeclampsia at 29-34 weeks of gestation were calculated using flow cytometry. The plasma ET-1 level was also determined using a modified radioimmunoassay. The plasma ET-1 concentrations and the Th1:Th2 cell ratios in normal pregnancies were significantly lower than those in patients with preeclampsia. Negative correlations were found between plasma ET-1 levels and Th2 cells in both the preeclamptic pregnancy groups and in the normal pregnant women. Our results indicate that elevated ET-1 levels are associated with a Th1:Th2 imbalance in preeclampsia.     

主动免疫治疗对不明原因习惯性流产患者辅助T细胞因子水平的影响

目的　探讨主动免疫治疗对不明原因习惯性流产 (UHA)患者辅助T细胞 (Th) 1 /Th2型细胞因子水平的影响。方法　采用酶联免疫吸附法 ,检测 30例半年内接受过淋巴细胞主动免疫治疗的UHA患者 (治疗组 ) ,及 2 5例未经治疗的UHA患者 (未治疗组 ) ,外周血单个核细胞 (PBMC)经滋养细胞抗原刺激产生的Th1型细胞因子白细胞介素 (IL) 2、γ干扰素 (IFN γ)及Th2型细胞因子产生IL 4、IL 1 0水平。并选取 1 5例正常非妊娠妇女作为对照 (对照组 )。结果　 (1 )在最佳诱导时间内 ,治疗组IL 2、IFN γ的水平分别为 (1 0 8± 37)ng/L、(1 1 0± 52 )ng/L ,明显低于未治疗组的 (2 2 3± 85)ng/L、(32 6±92 )ng/L(P值均 <0 .0 5) ;IL 4、IL 1 0水平分别为 (50± 1 1 )ng/L、(1 4 0± 37)ng/L ,明显高于未治疗组的(2 3± 1 1 )ng/L、(52± 2 8)ng/L(P值均 <0 .0 5)。未治疗组IL 2、IFN γ水平明显高于对照组的 (92± 32 )ng/L、(1 0 2± 35)ng/L(P值均 <0 .0 5) ;IL 4、IL 1 0水平低于对照组的 (62± 2 1 )ng/L、(1 50± 42 )ng/L(P值均 <0 .0 5)。治疗组与对照组各细胞因子水平比较 ,差异均无显著性 (P值均 >0 .0 5)。 (2 )治疗组30例患者治疗后半年内妊娠 2 6例 ,其中 8例自然流产 ,IL 2、IFN γ水平明显高于 1 8例妊娠     

Presence of CD30(+) and CD30L(+) cells in human placenta and soluble CD30 levels in cord blood are independent of maternal atopy

The intrauterine environment is characterized by a Th2 dominance during pregnancy, a milieu that also promotes atopic allergy. The aim of this study was to compare the presence of CD30, a molecule associated with Th2 related disorders such as atopic allergy, and its ligand (CD30L) in placenta in order to investigate if the placenta environment differs between atopic and non-atopic women. Serum concentrations of soluble CD30 (sCD30) from the mothers and their newborns were also elucidated. There were no differences in the immunohistochemical expression of CD30 and CD30L in placenta from atopic (n=28) compared with non-atopic (n=37) women. CD30 was expressed on the decidual stromal cells alone, while CD30L, previously not described in placenta, was detected on macrophage-like HLA-DR(+) cells throughout the mesenchymal chorionic villi. Serum sCD30 in atopic mothers was significantly elevated compared with serum sCD30 in non-atopic mothers (P< 0.05), while sCD30 levels in cord blood were similar in both groups independently of maternal atopic heredity. We suggest that sCD30 in cord blood and CD30 expression by decidual cells may reflect the Th2 environment surrounding the fetus, and both CD30 and CD30L could have immune regulatory functions in placenta.     

T辅助细胞1,2细胞因子平衡与正常妊娠及习惯性流产关系的 …

目的 探讨Ｔ辅助细胞１（Ｔｈ１）细胞因子（γ干扰素、肿瘤坏死因子α）、Ｔｈ２细胞因子（白细胞介素４、白细胞介素１０）平衡在母胎免疫耐受中的作用机理。方法 用半定量逆转录－聚合酶链反应（ＲＴ＿ＰＣＲ０技术,检测１１例正常未妊娠妇女、１０例正常早期妊娠妇女、１１例正常晚期妊娠妇女、１１例习惯性流产患者的外周血单个核细胞内γ干扰素、肿瘤坏死因子α、白细胞介素４、１０ｍＲＮＡ表达水平的相对含量。结果 （１     

Essential metal and immunoglobulin levels in women and neonates with PROM

The aim of the study was to evaluate the essential metals' levels in the blood of both mothers and neonates with premature rupture of the fetal membranes and to examine the changes produced by PROM in immunoglobulins' concentrations. Metals' levels in blood samples were determined by atomic absorption spectrophotometry, while the quantitative estimation of the immune proteins in both maternal and umbilical blood sera was performed by a single radial immunodiffusion. Zinc level in blood samples of mothers from the examined group was significant lower than this one evaluated in controls while the other concentrations of metals did not differ significant. We found the significant higher level of cord blood IgA in neonates from PROM+ group comparing with the PROM- group. Low zinc level in maternal blood which was found in examined PROM+ group might suggest that it constitutes one of the causes leading premature rupture of the fetal membranes. The response of the humoral immune system in PROM+ neonates seems to be stimulated by the premature rupture of the fetal membranes.     

Co-ordinate expression of Th1/Th2 phenotypes in maternal and fetal blood: evidence for a transplacental nexus

If maternal atopy and environmental exposure affect prenatal Th cell development, the maternal and fetal immune systems should display common Th1/Th2 phenotypes. To test this hypothesis, we studied maternal and neonatal blood samples from mothers with total serum IgE <300 IU/mL. Basal levels of IFN-γ, IL-4, and eotaxin in paired maternal and fetal sera were tightly correlated. Polyclonal T cell activation in vitro by Staphylococcal exotoxin B induced co-ordinate IFN-γ production from paired maternal and fetal mononuclear cells, accompanied by co-ordinate increases in activated CD4+CD69+ cells that display the CCR4+Th2 and CXCR3+ Th1 phenotypes. Maternal and fetal CD4+CXCR3+ T cells were subsequently identified as the major producers of IFN-γ. The data established that a transplacental nexus exists during normal pregnancy and that fetal Th cell responses may be biased by the maternal immune system.     

Role of T-cell cytokines in decidua and in cumulus oophorus during pregnancy

We focus on the roles of decidual and cumulus oophorus T cells. It was suggested that Th1-type cytokines (IFN-gamma and TNF-beta), which promote allograft rejection, may compromise pregnancy, whereas the Th2-type cytokines (IL-4, IL-5, IL-10) inhibiting the Th1 responses promote allograft tolerance and therefore may improve fetal survival. We found that T cells' leukemia inhibitory factor (LIF), M-CSF, IL-4 and IL-10 production at the fetomaternal interface could contribute to the maintenance of pregnancy. Interestingly, we did not find an increased production of IFN-gamma by decidual T cells during spontaneous abortion, as expected. We detected T cells in the cumulus oophorus, which surrounds the oocyte during ovulation and the egg before implantation. Cumulus oophorus T cells produce higher levels of IL-4 and LIF than the T cells of peripheral blood or the ovary. We can only speculate what the factors present in the microenvironment of the T cells are that could be responsible for the cytokine profile of decidual and cumulus oophorus T cells. Hormones present in the decidua and in the cumulus oophorus could be the first candidates. In particular, progesterone is a potent inducer of production of Th2-type cytokines, LIF and M-CSF. Other candidates could be molecules produced by the trophoblast or the embryo.     

Maternal circulating interferon-gamma and interleukin-6 as biomarkers of Th1/Th2 immune status throughout pregnancy.

AIM: T cells may be classified as T helper type 1 (Th1) cells, which synthesize cytokines inducing cellular immunity, or T helper type 2 (Th2), which synthesize cytokines inducing humoral immunity. According to the Th1/Th2 paradigm, it has been postulated that successful pregnancy induces an immune Th2 bias, but it is not yet clear how Th1 and Th2 systems vary simultaneously throughout the pregnancy. METHODS: Using maternal circulating interferon-gamma (IFN-gamma) and interleukin-6 (IL-6) as biomarkers of Th1 and Th2 cytokines, respectively, we examined the variation of circulating Th1/Th2 ratio in 35 healthy pregnant women from 10 to 40 weeks of pregnancy. RESULTS: With increasing gestational age, maternal circulating levels of IFN-gamma decrease, whereas those of IL-6 increase. The IFN-gamma/IL-6 ratio switches around the 19th week of pregnancy. CONCLUSIONS: Our results suggest that maternal systemic IFN-gamma and IL-6 concentrations may be biomarkers of Th1/Th2 immune status during pregnancy. Moreover, our findings showed that contrary to the Th1/Th2 paradigm, the Th1 bias may be prevailing at the beginning of pregnancy, balanced in the middle of pregnancy and supplanted by the Th2 bias at the end of pregnancy.     

复发性流产患者外周血T淋巴细胞亚群及CD_4~ 辅助性T淋巴细胞亚型的变化

目的　探讨不明原因的复发性流产 (URSA)与T淋巴细胞免疫功能的关系。方法　采用酶联免疫斑点 (ELISPOT)法对 30例URSA患者和 2 4例正常妇女 (对照 )的外周血T淋巴细胞亚群及CD 4 辅助性T淋巴细胞 (Th细胞 )亚型进行检测。结果　与对照相比 ,URSA患者外周血中CD 4 辅助性T淋巴细胞百分率增多 (分别为 45 %、6 0 % ,P <0 0 5 ) ,Th1细胞的百分率升高 (14%与 2 6 % ,P<0 0 1) ,Th2细胞的百分率降低 (2 0 %与 7% ,P <0 0 5 ) ,Th1/Th2细胞比值增大 (0 73与 3 80 ,P<0 0 1)。结论　URSA的发生可能与CD 4 辅助性T淋巴细胞的增多及Th1细胞介导的细胞免疫功能的增强有关。     

Cytokine patterns in maternal blood after premature rupture of membranes.

OBJECTIVE: To compare two types of cytokines, type 1, which activate cell-mediated reactions and are important in cytotoxic and delayed-type hypersensitivity reactions, and type 2, which encourage vigorous antibody production and are commonly found in association with humoral immune responses, in blood of women with premature rupture of membranes (PROM). METHODS: Forty-four women with histories of at least three successful pregnancies and who currently delivered normally served as controls. The PROM group consisted of 30 women with spontaneous rupture of fetal membranes at term. Peripheral blood mononuclear cells were stimulated separately with a mitogen, placental cells, and a trophoblast antigen extract, and the supernatants examined for type 1 and type 2 cytokines. RESULTS: Mitogen-stimulated blood cells produced significantly higher levels of type 1 cytokines in PROM women than in normal controls. Higher levels of the type 1 cytokine interferon-gamma were produced by PROM samples stimulated with autologous placental cells and with trophoblast antigens. Ratios of type 1 to type 2 cytokines were higher in PROM compared with normal pregnancy, and in some cases as much as 25-fold higher. CONCLUSION: Women in the PROM group had a stronger type 1 reactivity whereas normal women were more predisposed to type 2 immunity; thus, PROM appears to be associated with a maternal type 1 bias.