The Immunological Pregnancy Protective Effect of Progesterone Is Manifested via Controlling Cytokine Production

PROBLEM: This study was aimed at investigating the involvement of an altered cytokine pattern in the immunomodulatory and anti-abortive effects of a progesterone-induced immunomodulatory protein (PIBF). METHOD: PIBF expression on lymphocytes of healthy pregnant women and from women at risk for premature pregnancy termination was determined. In sera of the same women TNFα was quantified by a bioassay using L929 cells. NK activity was determined by a single cell cytotoxicity assay. Cytokine production of the lymphocytes or murine spleen cells was measured by ELISA or detected by immunocytochemistry. In pregnant mice endogenous PIBF activity was neutralized by anti-PIBF IgG. RESULTS: Sera of women at risk for premature pregnancy termination contained significantly higher concentrations of TNFα than those from healthy pregnant women and PIBF expression on the lymphocytes was inversely related to serum concentration of TNFα. Increased NK activity of lymphocytes after neutralization of endogenous PIBF activity is corrected by anti-IL2 treatment and PIBF inhibits IL12 expression on activated lymphocytes. PIBF increases IL-10 production by activated spleen cells. In pregnant mice, neutralization of endogenous PIBF activity by specific antibody results in increased resorption rate and reduced splenic IL-10 production. CONCLUSIONS: Our data allow the assumption that via blocking IL-12 production PIBF inhibits NK activation with a concomitant reduction of TNFα levels. Disturbances in this system might lead to the expression of the known synergistic effect of IL-12 and TNFα, resulting in a Th1 type cytokine dominance and pregnancy termination.     

The role of gamma/delta T-cell receptor-positive cells in pregnancy: part II.

PROBLEM: We have previously demonstrated a significantly increased ratio of gamma/delta T-cell receptor (TCR)-positive progesterone receptor(PR)-positive cells in the peripheral blood of healthy pregnant women compared to that of recurrent aborters or non-pregnant individuals. Treatment of pregnancy lymphocytes with a pan anti-gamma/delta TCR antibody inhibits progesterone-induced blocking factor (PIBF) production, increases natural killer (NK) activity, and alters the cytokine profile. The present study was aimed at investigating the role of the different gamma/delta subpopulations in these phenomena. METHOD OF STUDY: Peripheral blood lymphocytes from healthy pregnant women were incubated with either anti-gamma1.4 and delta1, or anti-gamma9 and delta2 antibodies. The effect of these treatments on PR induction and interleukin (IL)-10 and IL-12 expression were tested by immunocytochemistry. NK activity of anti-gamma/delta treated lymphocytes was also determined. RESULTS: In peripheral blood of healthy pregnant women, the most frequently occurring chain combination was gamma1.4/delta1, whereas in recurrent aborters, the gamma9/delta2 combination was predominant. Treatment of normal pregnancy lymphocytes with a mixture of gamma1.4 and delta1 antibodies resulted in a significantly reduced NK activity and increased PR and IL-10 expression, whereas treatment with a mixture of gamma9 and delta2 antibodies significantly reduced IL-10 production and slightly increased IL-12 production and NK activity. These data suggest the presence of two functionally distinct subpopulations in the peripheral blood of pregnant women.     

Progesterone regulates IL12 expression in pregnancy lymphocytes by inhibiting phospholipase A2

PROBLEM: Progesterone-induced blocking factor (PIBF) is one of the pathways that mediate the immunological effects of progesterone. PIBF inhibits natural killer (NK) cytotoxic activity. Recently we showed that neutralization of PIBF results in an increased interleukin (IL)-12 expression, which is corrected by cyclooxygenase inhibitors. As exogenous arachidonic acid (AA) voids the NK blocking effect of PIBF, it is likely that PIBF acts before the level of the cyclooxygenase enzyme. Therefore in this study we investigated the effect of PIBF neutralizing antibody and simultaneous phospholipase A2 inhibitor quinacrine (Q) treatment on IL-12 production. METHODS: Pregnancy lymphocytes were treated with anti-PIBF antibody or lipopolysaccharide (LPS) as a positive control, in the presence or absence of Q. IL-12 expression by PBMC was detected by immunocytochemistry. RESULTS: Neutralization of PIBF as well as LPS treatment resulted in an increased IL-12 expression, which was corrected by simultaneous Q treatment. Pre-treatment of lymphocytes with progesterone prevented the stimulating effect of LPS on IL-12 production. CONCLUSION: Progesterone binding of the lymphocytes is followed by the release of PIBF that inhibits AA release. The subsequent block of prostaglandin synthesis reduces IL-12 production and results in a lowered cytotoxic NK activity, which may contribute to a normal pregnancy outcome.     

The Expression of a Progesterone-Induced Immunomodulatory Protein in Pregnancy Lymphocytes

PROBLEM : The immunological effects of progesterone are mediated by a protein, named the progesterone-induced blocking factor (PIBF). The PIBF blocks NK activity in vitro and therefore prevents the abortive effect of high NK activity in mice. Increased NK activity has been suggested to play a role in pregnancy termination; thus NK inhibitory effect of the PIBF should contribute to the maintenance of normal gestation. This study was designed to investigate the relationship between in vivo PIBF-producing capacity and in vitro cytotoxic activity of pregnancy lymphocytes, as well as the clinical status or the outcome of pregnancy. METHOD : Lymphocytes of 168 pregnant women (96 normal pregnancies, 16 showing clinical symptoms of threatened preterm pregnancy termination, 46 recurrent aborters, and 10 women sampled at the onset of spontaneous abortion or preterm delivery) were isolated on Ficoll-Paque gradient. The lymphocytes were tested for reactivity with a PIBF-specific antibody by immunocytochemistry, and simultaneously for cytotoxic activity to human embryonic fibroblast targets. RESULTS : The percentage of PIBF-positive lymphocytes in peripheral blood of healthy pregnant women was significantly higher than in that of women at risk for premature pregnancy termination. In peripheral blood of patients undergoing spontaneous pregnancy termination at the time of sampling, and in those of women showing symptoms of premature pregnancy termination we found lower than normal percentage of PIBF-positive cells. PIBF expression of the lymphocytes showed an inverse correlation with NK activity, and the rate of PIBF positive lymphocytes was related to the outcome of pregnancy. CONCLUSION : These data suggest a strong relationship between PIBF producing capacity as well as NK activity of the lymphocytes and the success of gestation.     

The role of gamma/delta T cell receptor positive cells in pregnancy.

PROBLEM: Due to the lack of classical HLA antigens on the trophoblast, fetal antigens are possibly presented in a non major histocompatibility complex (MHC) restricted way. Decidual gammadelta T cells, which significantly increase in number during pregnancy, might play a role in recognition of fetal antigens and also in determining the quality of the response to these antigens. Our study was aimed at investigating the role of this cell population in progesterone-dependent immunomodulation. METHOD OF STUDY: Peripheral lymphocytes from healthy pregnant women and from habitual aborters were tested by immunocytochemistry for the presence of gamma/delta T cell receptor (TCR) and progesterone receptor. To investigate the effect of treatment with a pan anti gamma/delta antibody, lymphocytes were incubated for 3 hr with the antibody, and then interleukin (IL)-10, IL-12 and progesterone-induced blocking factor (PIBF) expression (by immuno-cytochemistry) as well as natural killer (NK) cell activity were determined. RESULTS: In peripheral blood of healthy pregnant women the percentage of gamma/delta TCR+ cells was significantly higher (P < 0.001) than in that of recurrent aborters or of non-pregnant individuals. Ninety-seven percent of gamma/delta TCR+ pregnancy lymphocytes expressed progesterone receptor. Binding of a specific antibody to the gamma/delta TCR inhibited PIBF- as well as IL-10 production, whereas it increased NK activity and IL-12 expression. CONCLUSIONS: These data suggest the role of gamma/delta TCR-bearing lymphocytes in progesterone-dependent immunomodulation.     

Progesterone as an immunomodulator

In the presence of progesterone, lymphocytes of healthy pregnant women produce a 34-kDa protein, which, because of its immunomodulatory effects has been called progesterone induced blocking factor (PIBF). PIBF is a secreted molecule and thus appears in biological fluids. Urinary PIBF concentrations in healthy pregnant women are significantly higher than in recurrent aborters, or in women showing clinical symptoms of threatened abortion. Recent data suggest, that PIBF production is not exclusively a pregnancy-related phenomenon. PIBF is present in most undifferentiated tissues, e.g. malignant tumors. The possible role of PIBF in tumor development will be discussed.     

The role of gamma/delta T cells in progesterone-mediated immunomodulation during pregnancy: a review.

PROBLEM: To determine if pregnancy is recognized by the immune system and if inadequate recognition of fetal antigens might result in failed pregnancy. METHOD OF STUDY: Review of literature and current data. RESULTS: In the decidua gamma/delta TCR positive cells significantly increase in number. A subset of gamma/delta T cells reacts with nonpolymorphic Class I or Class I like molecules. Trophoblast recognition is mediated by the V gamma 1 subset which recognize a conserved mammalian sequence on the trophoblast. Almost all gamma/delta T cells in the decidua are activated and use the V delta 1 chain, whereas the majority of human peripheral gamma/delta lymphocytes expresses V gamma 9/V delta 2 TCR. Peripheral gamma/delta T cells of healthy pregnant women preferentially use V gamma V delta 1 chains, on the other hand, those of recurrent aborters use the V gamma 9V delta 2 combination. Signaling via the V gamma 1.4V delta 1 receptor induces a Th2 type response, whereas activation of the lymphocytes via the V gamma 9V delta 2 receptor results in increased IL-12 production and natural killer (NK) activity. In the presence of progesterone, activated lymphocytes synthesize the progesterone induced blocking factor (PIBF), which inhibits NK activity and exerts an anti abortive effect in vivo. Decidual CD56+ and gamma delta+ cells are to a high extent the same population. CONCLUSION: All decidual CD56+ cells express PIBF, thus it cannot be excluded that local production of this substance contributes to low decidual NK activity and thus to the success of the pregnancy.     

Natural killer cell activity and cytokine production after in vitro immunoglobulin treatment of lymphocytes derived from pregnant women with or without risk for spontaneous abortion

OBJECTIVE: To investigate the possible mechanism of action effective in immunoglobulin G (IgG) treatment of recurrent spontaneous abortion (RSA). The effect in vitro of a commercially available intravenous immunoglobulin (IvIg) on the rate of interleukin (IL)-10 and IL-12 positive cells (Th1/Th2 balance) and on natural killer (NK) cell activity in populations of peripheral lymphocytes of healthy pregnant women and women at risk for premature pregnancy termination was studied. Primary habitual aborters as well as women showing clinical symptoms (bleeding or regular uterine contractions) of threatened premature pregnancy termination were included. METHODS: Lymphocytes of 20 pregnant women were tested. Five different batches of an IvIg with reported immunomodulatory potential were used at a concentration of 10 mg/mL. Cytokine profiles of the lymphocytes were determined by immunocytochemistry. For testing of NK cell activity, the 4 hr single cell cytotoxicity assay was used. RESULTS: Incubation with IgG of lymphocytes from recurrent spontaneous aborters concomitantly and significantly decreased the rate of IL-12 positive cells (P < 0.01) and increased the rate of IL-10 positive cells (P < 0.01), whereas such treatment had no significant effect on lymphocytes of pregnant women not at risk of abortion. Dialysis or heat treatment (56 degrees C, 30 min) of the IgG preparations did not modify the effect. Elevated NK cell activity of women at risk for premature pregnancy termination significantly decreased after IgG incubation of cells in all cases, whereas NK cell activity of normal pregnancy lymphocytes was not altered. CONCLUSION: This study suggests that incubation of peripheral lymphocytes from RSA patients with polyclonal polyspecific IgG alters cytokine profiles and NK activity while the same treatment does not affect lymphocytes of healthy pregnant women. These data might add to the understanding of mechanisms of action of IvIg in prevention of recurrent pregnancy loss.     

The impact of dydrogesterone supplementation on hormonal profile and progesterone-induced blocking factor concentrations in women with threatened abortion

PROBLEM: The therapeutic value of progestogens in threatened abortion is still under debate. In the presence of sufficient progesterone levels during pregnancy, lymphocytes synthesize a mediator [progesterone-induced blocking factor (PIBF)] that is anti-abortive in mice. The aim of this study was to evaluate the effect of dydrogesterone on pregnancy outcome of threatened aborters. METHOD OF STUDY: Twenty-seven threatened aborters were treated for 10 days with dydrogesterone (30-40 mg/day). Sixteen healthy pregnant controls received no treatment. Serum progesterone and estradiol concentrations as well as urine PIBF concentrations were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Pregnancy outcomes in dydrogesterone-treated threatened aborters did not statistically differ from those in healthy controls. Serum progesterone concentrations in control patients, but not those in threatened aborters increased as pregnancy progressed. Following dydrogesterone treatment, initially low PIBF concentrations of threatened aborters significantly increased (P = 0.001) to reach the PIBF level found in healthy controls. CONCLUSIONS: These data suggest that by inducing PIBF production, dydrogesterone might improve pregnancy success rates in threatened aborters.     

PIBF: The Double Edged Sword. Pregnancy and Tumor

Citation Szekeres‐Bartho J, Polgar B. PIBF: The Double Edged Sword. Pregnancy and Tumor. Am J Reprod Immunol 2010; 64: 77–86 Problem The role of progesterone‐dependent immunomodulation in the maintenance of normal pregnancy. Methods In vitro and in vivo data on the effect that progesterone and its mediator progesterone‐induced blocking factor (PIBF) exert on the immune functions of pregnant women are reviewed, together with clinical findings. Results Activated pregnancy lymphocytes express progesterone receptors, which enable progesterone to induce a protein called PIBF. PIBF increases Th2 type cytokine production by signaling via a novel type of IL‐4 receptor and activating the Jak/STAT pathway. PIBF inhibits phosholipase A2, thus reduces prostaglandin synthesis. PIBF inhibits perforin release in human decidual lymphocytes and reduces the deleterious effect of high NK activity on murine pregnancy. PIBF production is a characteristic feature of normal human pregnancy, and its concentration is reduced in threatened pregnancies. PIBF mRNA and protein are expressed in a variety of malignant tumors. Inhibition of PIBF synthesis increases survival rates of leukemic mice. Conclusion Progesterone‐induced blocking factor is produced by pregnancy lymphocytes and also by malignant tumors. The PIBF‐induced Th2‐dominant immune response is favorable during pregnancy but might facilitate tumor growth by suppressing local antitumor immune responses.     

Progesterone-Induced Blocking Factor and Cytokine Profile in Women with Threatened Pre-Term Delivery

Problem  The objective of this study was to compare serum concentrations of progesterone-induced blocking factor (PIBF), anti-inflammatory (IL-10), and pro-inflammatory (IL-6, TNFα, and IFNγ) cytokines of women with threatened pre-term delivery, with those of women with normal pregnancy and to evaluate the impact of PIBF on the outcome of pregnancy.
Method of study  A prospective study was conducted on a sample of 30 women with threatened pre-term delivery (study group) and 20 healthy pregnant women (control group) between the 24th and 37th gestational weeks. Serum PIBF, anti-inflammatory (IL-10), and pro-inflammatory (IL-6, TNFα, and IFNγ) cytokine concentrations were measured by enzyme-linked immunosorbent assay (ELISA).
Results  Thirteen of 30 patients (43.3%) with symptoms of threatened pre-term delivery, and one of 20 patients (5%) in the control group delivered before the 37th week of gestation. Mean PIBF concentrations in serum samples of patients with threatened pre-term delivery were significantly lower than in those of healthy pregnant women (171.12 ± 162.06 ng/mL versus 272.85 ± 114.87 ng/mL; P  < 0.05). Women with symptoms of threatened pre-term delivery had significantly lower serum levels of IL-10, and higher levels of IL-6 as well as IFNγ compared with healthy controls.
Conclusion  Our results indicate that measuring PIBF and cytokine concentrations in serum during pregnancy is feasible and may be important for understanding immunological causes of pre-term delivery.     

Progesterone Induced Blocking Factor Seen in Pregnancy Lymphocytes Soon After Implantation

PROBLEM: The immunomodulatory effect of progesterone (P) in pregnancy manifested via a protein named the P-induced blocking factor (PIBF) was previously reported. The goal of this study was to measure and compare the PIBF expression on lymphocytes between pregnant and non-pregnant women especially in early pregnancy. METHODS: PIBF expression was determined by immunocytochemistry using a PIBF-specific polyclonal antibody. Levels were assessed during the mid-cycle, luteal phase, and first trimester of pregnancy. RESULTS: PIBF expression was found in 24.9% of mid-cycle sera, 49% of luteal phase sera of women who failed to conceive, and 75% of luteal phase sera of women who conceived. CONCLUSIONS: These data indicate that the percentage of PIBF expressing lymphocytes increases as a result of pregnancy and that the stimulus for PIBF induction occurs soon after implantation. These data support the concept that PIBF may play an important role in early implantation possibly by inhibiting the destructive function of natural killer lymphocytes.     

Progesterone and Non-specific Immunologic Mechanisms in Pregnancy

PROBLEM: Progesterone-dependent immunomodulation is one of the mechanisms that enables pregnancy to proceed to term. Immunologic effects of progesterone are mediated by a protein named the progesterone-induced blocking factor (PIBF). Among other effects this protein inhibits natural killer (NK) activity and displays an anti-abortive effect in mice. Recently we have shown that PIBF induces a Th2 shift in vitro. The present study was aimed at investigating the in vivo effect of PIBF on cytokine production, as well as the relationship between cytokine production, NK activity, and pregnancy loss. METHOD OF STUDY: Balb-c mice on day 8.5 of pregnancy were injected intraperitoneally with 0.5 mg of rabbit anti-PIBF immunoglobulin G (IgG). Another group of mice was simultaneously treated with anti-NK monoclonal antibodies. Mice treated with the same amount of normal rabbit serum or untreated mice of similar gestational age were used as controls. The animals were sacrificed on day 10.5, and their uteri were inspected. The ratio of living and resorbed embryos was determined. NK activity as well as cytokine expression on the spleen cells were determined by immunocytochemistry and enzyme-linked immunoadsorbent assay (ELISA). RESULTS: Mitogen-activated spleen cells from anti-PIBF-treated mice produced significantly (P < 0.001) less IL-10 than those of pregnant control mice. A significantly higher percentage (P < 0.001) of spleen cells from anti-PIBF-treated mice expressed interferon-γ (IFNγ) as determined by immunocytochemistry, than those of untreated pregnant mice. There was a positive relationship between the percentage of IFNγ-positive spleen cells and resorption rates, and an inverse relationship between the latter and interleukin-10 (IL-10) production. All these effects were corrected by treatment with anti-NK antibodies. CONCLUSION: Our data suggest that PIBF contributes to the success of gestation via eytokine-mediated inhibition of NK activity.     

Expression of an Immunomodulatory Protein Known as Progesterone Induced Blocking Factor (PIBF) Does Not Correlate With First Trimester Spontaneous Abortions in Progesterone Supplemented Women

PROBLEM: An immunomodulatory protein known as the progesterone induced blocking factor (PIBF) has been found to positively correlate with early pregnancy beta human chorionic gonadotropin (B-hCG) levels. The study presented herein evaluated PIBF levels from conception to the end of the first trimester to determine if lower levels will correlate with first trimester spontaneous abortions (SAB). METHOD: Progesterone induced blocking factor expression by lymphocytes measured using an immunocytochemistry method was compared in pregnant women with ongoing vs. failed pregnancies. RESULTS: There were no differences in the proportion of women having lymphocytes expressing PIBF or in the median numbers when comparing ongoing vs. failed pregnancies. There was no B-hCG interval where failed pregnancies were found to have lower frequency of PIBF expressing lymphocytes. CONCLUSION: Inadequate PIBF expression independent of low P levels does not appear to be an etiologic factor for first trimester SABs; thus measuring this protein in pregnant women lacks practical usefulness.     

Changes in cytokine production during and after normal pregnancy

PROBLEM: The systemic T helper 1/T helper 2 (Th1/Th2) cytokine balance during normal human pregnancy is controversial, and observations about the balance in the postpartum period have only been reported for up to 3 months. METHOD: Whole-blood, from 83 healthy pregnant women, 80 healthy postpartum women, and 31 healthy non-pregnant women was stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin, and the levels of cytokines in the supernatant were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The production of all measured cytokines decreased during pregnancy, especially in the second trimester. After delivery, interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) increased from 2 to 11 months postpartum, and IL-4 increased from 6 to 11 months postpartum. CONCLUSIONS: These data indicate that 1) decreases in production of both Th1-and Th2-type cytokines during pregnancy may be related to the pregnancy-induced amelioration of autoimmune diseases: 2) increases in production of both Th1- and Th2-type cytokines in the postpartum period may be related to the postpartum aggravation of autoimmune diseases.     

The impact of dydrogesterone supplementation on serum cytokine profile in women with threatened abortion

PROBLEM: The role of increased Th1 cytokine expression in pregnancy failure has been questioned recently. The therapeutic value of progestogens in threatened abortion (TA) is still debated. The aim of this prospective study was to compare serum cytokine [tumor necrosis factor (TNF)-alpha, interleukin (IL)-12 and IL-10] concentrations in women with TA to those in women with normal pregnancy and to evaluate the impact of dydrogesterone supplementation in the former group on cytokine concentration. METHODS OF STUDY: Twenty-seven threatened aborters were treated for 10 days with dydrogesterone (30-40 mg/day). Sixteen healthy pregnant controls received no treatment. Serum cytokine concentrations were measured twice in both groups by enzyme-linked immunosorbent assay. RESULTS: Mean serum concentrations of Th1- and Th2-type cytokines in women with TA did not differ from those in women with normal pregnancy at first and second sampling. After dydrogesterone supplementation, mean TNF-alpha/IL-10 ratio changed from 1.08 to 1.75 while IL-12/IL-10 ratio remained almost the same (0.56-0.61) in the threatened aborters group and did not differ from those in healthy women. CONCLUSIONS: The results of this study indicate that peripheral cytokine production in threatened aborters does not differ from that observed among healthy pregnant women. The protective effect of dydrogesterone supplementation in threatened aborters is manifested via restoring progesterone-induced blocking factor concentration rather than controlling cytokine production.     

孕酮调节复发性流产患者淋巴细胞产生Th1、Th2因子及机制

目的探讨孕酮（P）对不明原因复发性自然流产（URSA）患者淋巴细胞产生Th1、Th2因子的调节作用及其机制。方法提取URSA患者的外周血单个核细胞（PBMC）,在植物血凝素（PHA）刺激下,分孕酮组和对照组培养后,采用ELISA法检测细胞培养上清中IL-12、IL-4的浓度,提取细胞全蛋白,用Western Blotting法检测过氧化物酶体增值物激活受体γ（PPAR-γ）的表达。结果孕酮组IL-12水平显著低于对照组（P〈0.01）,IL-4显著高于对照组（P〈0.05）。孕酮组PPAR-γ的表达显著高于对照组（P〈0.05）。结论孕酮能调节URSA妇女PBMC产生的Th1、Th2细胞因子,PPAR-γ可能参与了细胞因子表达的调控,PPAR-γ是重要的免疫调节因子。     

孕激素诱导的封闭因子在先兆流产患者血清中的表达

目的通过检测先兆流产患者血清中孕酮及孕激素诱导的封闭因子（PIBF）的表达水平,探讨PIBF与先兆流产的关系及临床意义。方法先兆流产患者72例为观察组,同期健康早孕妇女40例为对照组,采用酶联免疫吸附方法（ELISA）检测血清中孕酮及PIBF水平并进行对比分析。结果两组孕妇的年龄、妊娠时间和血清孕酮水平比较差异无统计学意义（P〉0.05）;观察组PIBF水平为（103.91±25.12）ng/ml,明显低于对照组的（117.62±24.33）ng/ml,差异有统计学意义（P〈0.05）。观察组血清孕酮水平与PIBF之间不存在相关关系（P〉0.05）,对照组血清孕酮水平与PIBF之间存在相关关系（P〈0.05）。结论 PIBF表达降低与自然流产的发生有关系,妊娠早期血清PIBF水平降低是引起自然流产的原因之一。     

Leptin, IL-10 and inflammatory markers (TNF-alpha, IL-6 and IL-8) in pre-eclamptic, normotensive pregnant and healthy non-pregnant women

PROBLEM: Despite progress in immunobiology, pre-eclampsia (PE) remains one of the most common reasons for women to die during pregnancy. The widespread pathophysiological mechanisms are endothelial dysfunction, oxidative stress and inflammation. The aim of this study was to assess the alteration in the levels of leptin, interleukin (IL)-10 and inflammatory cytokines [tumour necrosis factor (TNF)-alpha, IL-6 & IL-8] in pre-eclamptic (severe and mild), healthy pregnant and non-pregnant women and correlate these parameters with disease severity. METHOD OF STUDY: The levels of leptin, IL-10 and inflammatory cytokines were measured by high sensitivity enzyme-linked immunoabsorbant assay. The study subjects were 54 pre-eclamptic women (ten severe and 45 milder), compared by age matched 50 healthy pregnant and 27 non-pregnant women. Kruskal-Wallis non-parametric analyses of variance followed by Mann-Whitney U-test were used for statistical analysis. RESULTS: The levels of leptin, TNF-alpha, IL-6 & IL-8 in pre-eclamptic subjects were increased significantly when compared with the healthy control pregnant and non-pregnant (P < 0.000). The concentration of IL-10 has shown different pattern as its level decreased significantly (0.001) in pre-eclamptic women (overall) in comparison with control subjects (pregnant & non-pregnant). A combination of 80% or higher sensitivity and specificity was seen in the parameters analysed, except IL-8 and IL-10. CONCLUSION: Our findings suggest a relationship among TNF-alpha, IL-6, IL-8, IL-10 and leptin and indicate that altered levels of above markers in PE might be used as markers of pro-inflammation/anti-inflammation and endothelial dysfunction in pre-eclamptic pregnancies. These results also advocate the abnormal leptin and cytokine responses in mother, which might be involved in the pathogenesis of PE.     

Peripheral blood mononuclear cells stimulate progesterone production by luteal cells derived from pregnant and non-pregnant women: possible involvement of interleukin-4 and interleukin-10 in corpus luteum function and differentiation

Human luteal cells have been reported to express human leukocyteantigen-DR and lymphocyte functional antigen-3 on the cell surface,suggesting physiological interaction between luteal cells andT-lymphocytes through the menstrual cycle into early pregnancy.To elucidate the role of peripheral lymphocytes on corpus luteumdifferentiation, the effect of peripheral blood mononuclearcells (PBMC) on steroidogenesis by luteal cells was investigated.The production of Th-2 cytokines such as interleukin (IL)-4and IL-10 by the co-cultured cells was also examined, and theeffects of these cytokines on progesterone production by lutealcells were investigated. Corpora lutea were obtained from eightnon-pregnant women in the luteal phase and five women in earlypregnancy for luteal cell culture. PBMC were isolated from unrelatedwomen in the follicular phase, secretory phase, and early pregnancy.After co-culture with allogenic PBMC for 48 h, progesteroneproduction was significantly enhanced by PBMC from the secretoryphase and early pregnancy in the non-pregnant luteal cell culture.In the pregnant luteal cell culture, a significant increasein progesterone production was also observed by the co-culturewith PBMC from women in early pregnancy, showing that PBMC havea luteotrophic effect. The stimulatory effects of PBMC werealso observed in co-culture conditions which prevented directcell-to-cell interaction with luteal cells, showing the minorinfluence of mixed lymphocyte reaction. By co-culture with PBMC,the production of IL-10, but not IL-4, was significantly augmentedin luteal cell culture derived from non-pregnant women, whereasthe production of both IL-4 and IL-10 was significantly enhancedin the luteal cell culture derived from pregnant women. Moreover,IL-4 and IL-10 promoted progesterone production by culturedluteal cells, especially in the luteal cell culture derivedfrom corpora lutea of early pregnancy. These findings indicatethat PBMC stimulate progesterone production by luteal cellsand suggest the involvement of PBMC in corpus luteum functionand differentiation probably via the Th-2-type lymphocytes.