Elevated soluble Fas ligand levels may suggest a role for apoptosis in women with endometriosis

OBJECTIVE: To evaluate soluble Fas ligand concentrations in serum and peritoneal fluid from women with endometriosis and from fertile controls without endometriosis, and to study levels of soluble Fas ligand in conditioned media of cultured endometrial stromal cells. DESIGN: Prospective, experimental trial. SETTING: Two academic IVF centers. PATIENT(S): Twenty-nine fertile women without endometriosis and 57 infertile women with endometriosis (32 with stage I or II disease and 25 with stage III or IV disease). MAIN OUTCOME MEASURE(S): Enzyme-linked immunosorbent assay was used to measure soluble Fas ligand concentrations in paired samples of serum and peritoneal fluid from women with and without endometriosis. Concentrations were also measured in conditioned media of cultured endometrial stromal cells at basal conditions and after stimulation with interleukin-8 (0.001-10 ng/mL) and tumor necrosis factor-alpha (1-10 ng/mL). RESULT(S): Compared with fertile controls and women with early-stage of endometriosis, women with moderate to severe endometriosis had elevated serum (87.2 +/- 6.4, 88.2 +/- 6.9, and 162.3 +/- 7.8 pg/mL, respectively) and peritoneal fluid (81.0 +/- 6.0, 80.5 +/- 6.8, and 166.2 +/- 10.3 pg/mL, respectively) concentrations of soluble Fas ligand. Serum levels of soluble Fas ligand positively correlated with levels in peritoneal fluid. Comparison of patients in the same menstrual cycle in each group revealed that increased levels of soluble Fas ligand in patients with advanced endometriosis were not attributable to the difference in cycle phases. Soluble Fas ligand was not detected in conditioned media of endometrial stromal cells under baseline conditions or after stimulation. CONCLUSION(S): Serum and peritoneal fluid of women with moderate to severe endometriosis contain elevated concentrations of soluble Fas ligand compared to women with minimal or mild endometriosis and women without endometriosis. These findings suggest a role for apoptotic dysregulation in the pathophysiology of endometriosis.     

Role of B7-1 in mediating an immune response to myeloid leukemia cells

A costimulatory signal from B7-1 (CD80) to its counter-receptor CD28 is required for T-cell activation. Many tumors, including most human leukemias, lack expression of B7-1, and this has been suggested to contribute to the failure of immune recognition of these diseases. A murine leukemia model system was developed to assess the potential role of B7-1 in the induction immunity to leukemia cells. The nonleukemic 32Dc13 myeloid cell line was transformed by transfection of the BCR/ABL gene, generating a subline (32Dp210/clone 26) that was leukemic and rapidly lethal to syngeneic, immunocompetent C3H/HeJ mice or T-cell- deficient nude mice. B7-1-modified leukemic cells remained lethal in nude mice, but caused only a transient, nonlethal leukemia in C3H/HeJ mice. After a single exposure to live, nonirradiated B7-1-modified leukemic cells, C3H/HeJ mice developed protective immunity against subsequent challenge with B7-1(-) leukemic cells. Further, hyperimmunization with B7-1(+) leukemic cells prolonged the survival of mice previously injected with a lethal number of B7-1(-) leukemic cells. These results indicate that myeloid leukemic cells may be attractive candidates for B7-1 gene transfer.     

The effects of maternal haemoglobin as an indicator of maternal nutritional status on,maternal measles antibodies of mother-infant pairs at birth

Background

Maternal measles antibodies (MMA) are actively transferred through the placenta from mother to foetus. A relationship could exist between MMA of mother-infant pairs and maternal nutritional indicator (haemoglobin).

Objectives

This study reviewed the effects of maternal haemoglobin (Hb) on MMA of mother-infant pairs at birth.

Methods

One hundred and fifty three mother-infant pairs were enrolled in this study using the systematic random sampling method. Means of maternal Hb and MMA of mother-infant pairs were compared using the Student t test. Correlation coefficients of maternal Hb and MMA of mother-infant pairs were also determined. Multivariate analysis of variable (MANOVA) and covariates (MANCOVA) was used to investigate the effects of maternal Hb (fixed factor), gestational age, maternal age, birth weight (covariates) on combined MMA of mother-infant pairs (dependent factors). Benferroni adjusted Univariate linear regression was used to investigate the dependent variables separately.

Results

There were 78 (51%) males and 75 (49%) females. The (mean ± SD) MMA of mother-infant pairs at birth were 134.66 ± 93.31 (95% CI, 119.76 – 149.56) U/ml, and 187.49 ± 85.01 (95% CI, 173.91 – 201.07) U/ml, and their correlation was significant (p = 0.025). Ninety one (59.5 %) mothers had low Hb, 62 (40.5 %) had acceptable Hb levels. The overall mean maternal Hb was 11.01 ± 1.00 (95% CI, 10.85 – 11.17) g/dl . A positive significant correlation was observed between maternal Hb and MMA of the newborn-infant (p = 0.031). The MANOVA showed a statistically significant difference between maternal Hb on the combined dependent variables (p =0.033); however, results for the dependent variables using the Benferroni adjusted Univariate analysis was significant for only MMA of the infants, (p = 0.009).

Conclusion

There was a significant association between aacceptable levels of maternal Hb and high MMA of the newborn-infants. Therefore, these newborn infants start out with higher MMA that could give them better protection against measles during infancy.     

Mechanisms of chorioamnionitis‐associated preterm birth: interleukin‐1β inhibits progesterone receptor expression in decidual cells

In chorioamnionitis (CAM), a major cause of preterm birth (PTB), maternal–fetal inflammation of the decidua and amniochorion cause the release of cytokines that elicit cervical ripening, fetal membrane rupture and myometrial activation. We posit that this inflammatory milieu triggers PTB by inhibiting progesterone receptor (PR) expression and increasing decidual prostaglandin (PG) production. Immunohistochemical staining of decidua detected significantly lower PR levels in decidual cells (DCs) from CAM‐complicated PTB. Incubation of DCs with IL‐1β decreased PR expression and significantly increased PGE₂ and PGF_2α production and COX‐2 expression. The addition of PGF_2α to DC cultures also suppressed PR expression. However, the COX inhibitor, indomethacin, did not reverse IL‐1β suppression of PR expression in DC cultures. Although IL‐1β treatment activated the NF‐K B, ERK1/2 and p38 MAPK signalling cascades in DCs, inhibition of ERK1/2 MAPK signalling alone was sufficient to completely reverse the suppression of PR levels by IL‐1β. These findings suggest that CAM‐associated PTB is induced at least in part by IL‐1β‐mediated functional progesterone withdrawal. Copyright © 2015 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Seroprevalence of cytomegalovirus among pregnant women attending Murtala Mohammed Specialist Hospital Kano,Nigeria

Background

Primary Cytomegalovirus (CMV) infection during pregnancy is a frequent and serious threat to the fetus. As there is no vaccine alternative measures are needed to prevent congenital CMV infection.

Objective

This study determined CMV Immunoglobulin G (IgG) antibody among pregnant women in order to ascertain the immune status of mothers to guide policy makers.

Methods

A semi-structured questionnaire was initially administered to obtain information on demographic details, stage of pregnancy and risk factors. Blood was collected by venipuncture from 180 women attending the antenatal clinic in Murtala Mohammed Specialist Hospital Kano, Kano State, Nigeria. Sera samples were screened using CMV IgG ELISA kit (Dialab, Austria).

Results

Out of 180 pregnant women, 164 (91.1%) were seropositive. Based on stages of pregnancy 6/6(100%), 52/60(86.7%) and 106/114(93.0%) were seropositive among women in the first, second and third trimesters respectively.

Conclusion

Seroprevalence of pregnant women to CMV Ig G is high, hence the need for CMV - IgM screening to know the extent of active infection. There is also need for public enlightenment on the methods of transmission, effective prevention and control strategies.     

Interferon-γ Protects First-Trimester Decidual Cells against Aberrant Matrix Metalloproteinases 1, 3, and 9 Expression in Preeclampsia

Human extravillous trophoblast (EVT) invades the decidua via integrin receptors and subsequently degrades extracellular matrix proteins. In preeclampsia (PE), shallow EVT invasion elicits incomplete spiral artery remodeling, causing reduced uteroplacental blood flow. Previous studies show that preeclamptic decidual cells, but not interstitial EVTs, display higher levels of extracellular matrix–degrading matrix metalloproteinase (MMP)-9, but not MMP-2. Herein, we extend our previous PE-related assessment of MMP-2 and MMP-9 to include MMP-1, which preferentially degrades fibrillar collagens, and MMP-3, which can initiate a local proteolytic cascade. In human first-trimester decidual cells incubated with estradiol, tumor necrosis factor-α (TNF-α) significantly enhanced MMP-1, MMP-3, and MMP-9 mRNA and protein levels and activity measured by real-time quantitative RT-PCR, ELISA, immunoblotting, and zymography, respectively. In contrast, interferon γ (IFN-γ) reversed these effects and medroxyprogesterone acetate elicited further reversal. Immunoblotting revealed that p38 mitogen-activated protein kinase signaling mediated TNF-α enhancement of MMP-1, MMP-3, and MMP-9, whereas IFN-γ inhibited p38 mitogen-activated protein kinase phosphorylation. Unlike highly regulated MMP-1, MMP-3, and MMP-9, MMP-2 mRNA and protein expression was constitutive in decidual cells. Because inflammation underlies PE-associated shallow EVT invasion, these results suggest that excess macrophage-derived TNF-α augments expression of MMP-1, MMP-3, and MMP-9 in decidual cells to interfere with normal stepwise EVT invasion of the decidua. In contrast, decidual natural killer cell–derived IFN-γ reverses such TNF-α–induced MMPs to protect against PE.Preeclampsia (PE) is a multifactorial disease that affects 6% to 8% of pregnancies in the United States, is responsible for nearly 8% of maternal deaths, and is a leading cause of perinatal morbidity and mortality. Severe PE is a major indication for early, medically indicated preterm birth.¹ The diagnosis of PE is usually made after 20 weeks by the appearance of hypertension and proteinuria (maternal syndrome).¹During the first 20 weeks of gestation, extravillous trophoblasts (EVTs) arise from cytotrophoblast at the tips of placental anchoring villi and invade the decidua and upper third of the myometrium. As they navigate through the decidua, EVTs enter and facilitate remodeling of spiral arteries and arterioles into large-bore, low-resistance vessels that increase uteroplacental blood flow to the intervillous space requisite for fetal growth and development.^2,3 The onset of PE is strongly associated with shallow decidual EVT invasion, which leads to incomplete vascular transformation and reduced uteroplacental blood flow. The resulting hypoxic placenta⁴ secretes several putative inducers of endothelial cell activation and angiogenesis (eg, soluble flt-1 and endoglin) into the maternal circulation that elicits vascular damage,^5,6 leading to the maternal syndrome.¹Invasion of the decidua by EVT involves sequential attachment to adhesion molecules, followed by their degradation. Relevant integrin (ITG) heterodimers include ITG-α1/ITG-β1 and ITG-α5/ITG-β1, which recognize laminin/collagen IV and fibronectin, respectively, in the decidual extracellular matrix (ECM),^7–9 as well as vascular endothelial cadherin, an endothelial cell receptor.¹⁰ In addition to newly synthesized basement membrane–type proteins, the decidual ECM also contains significant residual interstitial collagens.¹¹ Degradation of the ECM scaffolding structure is mediated principally by matrix metalloproteinases (MMPs), a family of zinc-requiring enzymes that includes collagenases, gelatinases, and stromelysins.¹² Tissue inhibitors of MMPs (TIMPs) regulate MMP catalytic activity.¹³ The MMPs act in concert with urokinase-type plasminogen activator (uPA) and its specific inhibitor, plasminogen activator inhibitor-1 (PAI-1).¹⁴Previously, our laboratory compared immunostaining of the decidua from women with PE versus gestational age–matched control decidua for the presence of the basement membrane–degrading gelatinases, MMP-2 and MMP-9, as well as their respective inhibitors, TIMP-1 and TIMP-2, and found that PE is accompanied by a significant increase in MMP-9 levels in decidual cells, but not in interstitial EVTs. Unlike MMP-9, no PE-related changes in immunostaining were observed for either MMP-2 or TIMP-1 or TIMP-2 in either decidual cells or interstitial EVTs.¹⁵ Significant subsets of PE are associated with underlying maternal infections and/or inflammation,¹⁶ accompanied by an excess of decidual macrophages^17–20 that are likely sources of elevated levels of the proinflammatory cytokines IL-1β and tumor necrosis factor-α (TNF-α).²¹Consistent with the in situ observations described above and strong evidence that the pathogenesis of most cases of PE are initiated in early pregnancy,¹ we found that incubation of primary leukocyte-free, first-trimester human decidual cells with either IL-1β or TNF-α markedly enhanced MMP-9 mRNA and protein expression, unaccompanied by significant changes in either MMP-2 or TIMP-1 or TIMP-2 mRNA and protein expression.¹⁵The current study extends our previous PE-related assessment of MMP-2 and MMP-9 to include MMP-1, which preferentially degrades fibrillar collagens, and MMP-3, which can initiate a local proteolytic cascade by degrading a wide array of ECM proteins and by activating the secreted zymogenic form of other MMPs, such as pro–MMP-1 and pro–MMP-9.^13,22 We found the following using a two-tiered approach of integrating in situ with in vitro observations: immunoreactive MMP-1 and MMP-3 levels were compared in decidual cells and interstitial EVTs of decidual placental sections from women with PE versus gestational age–matched controls: MMP-1, MMP-3, as well as MMP-2 and MMP-9, were measured in the conditioned medium of primary, leukocyte-free, first-trimester human decidual cells incubated in parallel with estradiol (E₂), which was used as the control incubation for E₂ + medroxyprogesterone acetate (MPA) to mimic the pregnant steroid milieu. The steroids were added alone or with either TNF-α or interferon γ (IFN-γ) or TNF-α + IFN-γ. Inclusion of IFN-γ, a primary decidual natural killer (dNK) cell product,²³ was prompted by our recent observations that co-incubation of first-trimester human decidual cells with IFN-γ and either IL-1β or TNF-α synergistically enhances expression of two chemokines, interferon gamma-induced protein 10 (IP-10; alias CXCL10) and interferon-inducible T cell alpha chemoattractant (ITAC; alias CXCL11), that can selectively recruit the peripheral C-X-C chemokine receptor 3–expressing CD56^bright CD16(−) NK cell population to the decidua,²⁴ where they mediate several pregnancy protective effects.^25–27     

MMPs／TIMPs在甲状腺乳头状癌细胞中的表达与作用的研究

目的：探讨甲状腺乳头状癌（papillary thyroid carcinoma，PTC）患者癌细胞中基质金属蛋白酶（matrix metalloproteinases，MMPs）和金属蛋白酶的组织抑制剂（tissue inhibitor of metalloproteinases，TIMPs）蛋白的表达水平，探讨MMPs/TIMPs与甲状腺乳头状癌发生、发展之间的相互关系。方法：应用ELISA方法检测甲状腺正常组织和乳头状癌癌细胞培养液中MMP-2、MMP-9以及TIMP-1、TIMP-2的分泌水平：采用MTS掺人试验检测在rhMMP-2或抗MMP-2中和抗体的作用下，甲状腺正常组织细胞和乳头状癌癌细胞增殖水平。结果：甲状腺乳头状癌癌细胞分泌MMP-2、MMP-9水平提高，而分泌TIMP-1、TIMP-2水平降低，MMPs与TIMPs的比值升高特别显著；MMP-2对甲状腺乳头状癌癌细胞有明显的促增殖作用，抗MMP-2单克隆抗体能明显地抑制癌细胞生长。结论：MMPs与TIMPs的比值有望作为预测甲状腺乳头状癌生长和转移的一项检测指标。