Circulating ficolin-2 and ficolin-3 in normal pregnancy and pre-eclampsia.

Ficolins are soluble molecules of the innate immune system that recognize carbohydrate molecules on microbial pathogens, apoptotic and necrotic cells. They act through two distinct routes: initiating the lectin pathway of complement activation and mediating a primitive opsonophagocytosis. In this study, we measured plasma levels of ficolin-2 and ficolin-3 in 60 pre-eclamptic patients, 60 healthy pregnant women and 59 healthy non-pregnant women by enzyme-linked immunosorbent assay (ELISA). Circulating levels of complement activation products (C4d, C3a, SC5b9), angiogenic factors (soluble fms-like tyrosine kinase-1, placental growth factor) and markers of endothelial activation (von Willebrand factor antigen), endothelial injury (fibronectin) and trophoblast debris (cell-free fetal DNA) were also determined. Plasma levels of ficolin-2 were significantly lower in healthy pregnant than in healthy non-pregnant women, while ficolin-3 levels did not differ significantly between the two groups. Furthermore, pre-eclamptic patients had significantly lower ficolin-2 and ficolin-3 concentrations than healthy non-pregnant and pregnant women. In the pre-eclamptic group, plasma ficolin-2 levels showed a significant positive correlation with serum placental growth factor (PlGF) concentrations and significant inverse correlations with serum levels of soluble fms-like tyrosine kinase-1 (sFlt-1), blood urea nitrogen and creatinine, serum lactate dehydrogenase activities, as well as with plasma VWF:antigen, fibronectin and cell-free fetal DNA concentrations. In conclusion, circulating levels of ficolin-2 are decreased in the third trimester of normal pregnancy. There is a further decrease in plasma ficolin-2 concentrations in pre-eclampsia, which might contribute to the development of the maternal syndrome of the disease through impaired removal of the trophoblast-derived material released into the maternal circulation by the hypoxic and oxidatively stressed pre-eclamptic placenta.

OS004. A link between the complement system and angiogenic imbalance inpreeclampsia: ficolin-2 deficiency.

INTRODUCTION: An excessive maternal systemic inflammatory response to pregnancy, as well as an imbalance between circulating angiogenic factors and their antagonists plays a central role in the pathogenesis of preeclampsia. The complement system, as part of innate immunity, is fundamental to the host's immune defense against microbial pathogens, apoptotic and necrotic cells. Both of its excessive activation and deficiencies can lead to various disorders. OBJECTIVES: The aim of this study was to determine circulating levels of components of the complement system and their relationship to those of angiogenic factors in normal pregnancy and preeclampsia. METHODS: Sixty preeclamptic patients, 60 healthy pregnant women and 59 healthy non-pregnant women were involved in this case-control study. Circulating levels of C1rC1sC1-inh, C3bBbP, C4d, C3a, SC5b9, ficolin-2, ficolin-3, soluble fms-like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF), as well as activity of the complex of mannose-binding lectin and mannose-binding lectin-associated serine protease 2 (MBL-MASP2 complex) were measured. For statistical analyses, non-parametric methods were applied. RESULTS: Circulating levels of C3bBbP, C4d, C3a, SC5b9, sFlt-1, PlGF, as well as MBL-MASP2 activity were significantly higher, while ficolin-2 concentrations were significantly lower in healthy pregnant than in healthy non-pregnant women. Furthermore, preeclamptic patients had significantly higher C1rC1sC1-inh, C3bBbP, C4d, C3a, SC5b9 and sFlt-1 levels and significantly lower ficolin-2, ficolin-3 and PlGF concentrations than healthy pregnant women. In the groups of healthy pregnant women and preeclamptic patients, plasma ficolin-2 levels showed a significant positive correlation with serum PlGF concentrations and a significant inverse correlation with serum levels of sFlt-1. There was no other relationship between complement components and angiogenic factors in either study group. CONCLUSION: Elevated levels of activation products in the systemic circulation indicate complement activation with increased terminal complex formation in preeclampsia, which seems to be independent from alterations in circulating angiogenic factors. Nevertheless, low ficolin-2 concentrations might contribute to the angiogenic imbalance in preeclampsia by impaired removal of the sFlt-1-containing trophoblast-derived material released into the maternal circulation by the hypoxic and oxidatively stressed preeclamptic placenta.

OS085. Decreased maternal circulating PLGF is a significant predictor of length of pregnancy in women with hypertensive disorders of pregnancy.

INTRODUCTION: Diagnosis of the presence of disease and prediction of the rate of progression of disease in women with hypertensive disorders of pregnancy remains a clinical problem. Better methods are needed to determine the magnitude of risk to support patient counseling and clinical management. OBJECTIVES: To investigate whether the level of free PlGF is a significant predictor of length of pregnancy in women with hypertension. METHODS: In this case-control study a single sample was taken between the 22nd and 34th completed gestational weeks from 130 pregnant women with a final diagnosis of: pre-eclampsia (PE), HELLP-syndrome, superimposed pre-eclampsia (SIPE), chronic hypertension (CHT), gestational hypertension (GHT), and normal healthy pregnancy (Control). Plasma was analysed for PlGF using the Triage® PlGF assay (Alere, San Diego). A positive PlGF test was defined as below the 5th centile of normal healthy pregnancy. Hazard ratios for length-of-pregnancy were calculated for a positive PlGF test in a multivariate Cox proportional hazards model adjusting for two covariates, the gestational age at sample collection and a final diagnosis of proteinuric hypertension (PE, HELLP, and SIPE). RESULTS: Median PlGF concentration was significantly lower in women with hypertension than in controls. Women with proteinuric hypertension had the lowest levels of PlGF. A positive PlGF test predicted delivery before 35 weeks in 93.7% women, and delivery before 37 weeks in 90.5% women. A positive PlGF test was associated with a significantly higher risk of imminent delivery. PlGF was a significant and independent predictor of women destined to deliver early because of maternal or fetal complication (adjusted Hazard Ratio of 3.43, 95%CI of 1.97 to 5.98). CONCLUSION: A positive PlGF test is significant predictor of length of pregnancy, independent of other diagnostic criteria. PlGF has the potential to identify increased risk without the limitation of non-specificity which exists with other diagnostic parameters.

PP005. Natriuretic peptide precursor B gene (TTTC)N microsatellite polymorphism and elevated BNP levels in early onset pre-eclampsia.

INTRODUCTION: There is a variable tandem repeat (TTTC) polymorphism in the 5(')-flanking region of the natriuretic peptide precursor B gene (NPPB), which shows association with severe pre-eclampsia. The 11-repeat carrier frequency is significantly higher in severe pre-eclamptic patients. The 11/11 genotype carriers in the pre-eclamptics has significantly higher BNP levels. OBJECTIVES: Our aim was to identify this polymorphism in samples of early onset pre-eclamptic (EOP) patients, late onset pre-eclamptic patients and healthy controls. We also compared the natriuretic peptide B (BNP) concentrations. METHODS: Blood samples were collected from healthy pregnant normotensive women (n=235) and women with pre-eclampsia (n=220). DNA was isolated and fluorescent PCR and DNA fragment analysis was performed for the detection of (TTTC) repeats. The plasma BNP concentration was measured by fluorescence immunoassay method using Triage BNP (Alere, San Diego). RESULTS: We detected 12 different repeats on the NPPB gene. The overall distribution of alleles and genotypes was significantly different between the control and pre-eclamptic groups. The 11/11 genotype carriers showed a significantly higher frequency in early onset pre-eclamptic patients than in the healthy pregnant controls (p=0.026). Calculated odds ratio (OR) was 1.694 (95% CI: 1.06-2.70). In contrast the 11/11 genotype carrier frequency was not significantly higher in the late onset pre-eclamptic group than in the control group (p=0.5308), adjusted OR 1.22 (95% CI: 0.7138-2.086). In the early onset pre-eclamptics the 11/11 genotype showed a significantly higher frequency than in the late onset group (p=0.0039) OR: 6.00. The concentration of the BNP was 9.75pg/ml in the healthy controls and 32.40pg/ml in the pre-eclamptic group (p<0.0001). The 11/11 genotype carriers had significantly higher BNP levels in the pre-eclamptic group. The early onset pre-eclamptic patients had a significantly higher BNP concentration (40.55pg/ml) than in the late onset pre-eclamptic patients (24.1pg/ml) (p=0.013). CONCLUSION: The NPPB gene (TTTC) microsatellite polymorphism in the 5(')-flanking region showed a significant difference in the distribution of alleles and genotypes between early onset and late onset pre-eclamptic patients in an ethnically homogeneous population. The concentration of the BNP was higher in early onset pre-eclamptic women, and it showed association with the genotypes.

PP056. Placental growth factor is a better predictor of preterm birth than uterine or umbilical artery doppler in hypertensive disorders of pregnancy.

INTRODUCTION: Diagnosis of the presence of disease and prediction of the rate of progression of disease in women with hypertensive disorders of pregnancy remains a clinical problem. OBJECTIVES: Adequate placentation and placental development are important for normal pregnancy. We determined whether PlGF is a better predictor of delivery before 34+0 and 37+0 weeks than uterine artery and umbilical artery doppler. METHODS: One hundred and four women presenting before the completed 34th week of pregnancy with hypertensive disorders of pregnancy were enrolled into the study. Final diagnosis was chronic hypertension (N=24), gestational hypertension (N=21), pre-eclampsia (N=24), HELLP-syndrome (N=15), superimposed preeclampsia (N=20). Blood draw occurred before the 34th week of pregnancy at the time of investigation for expedited delivery or expectant management. Plasma was analysed for PlGF by the Alere Triage® PlGF assay using fluorescently-labelled monoclonal antibodies against PlGF. A positive PlGF test was defined as below the 5th centile of normal healthy pregnancy. RESULTS: Of the 104 pregnant women, the level of PlGF was abnormal in 23 of 24 (96%) women with IUGR, compared to 10 of 24 (42%) and 14 of 24 (58%) for uterine artery doppler and umbilical artery doppler, respectively. In the case of pre-eclampsia, the level of PlGF was abnormal in 50 of 59 (85%), compared to 15 of 59 (25%) and 25 of 56 (45%) for uterine artery doppler and umbilical artery doppler, respectively. Forty four (42%) women required medical delivery before 34+0 weeks gestation and 68 (65%) before 37+0 weeks gestation (see Table). PlGF detected a higher number of women requiring early delivery than doppler. CONCLUSION: The new Triage® PlGF test provides useful information to inform clinical decisions in pregnancy-associated hypertensive disorders, before the 34th completed gestational week.

Association of elevated serum heat-shock protein 70 concentration with transient hypertension of pregnancy, preeclampsia and superimposed preeclampsia: a case-control study.

Our aim was to investigate the association between serum heat-shock protein (Hsp) 70 concentration and hypertensive disorders of pregnancy. One hundred and forty-two pregnant women with hypertensive disorders (93 with preeclampsia, 29 with transient hypertension of pregnancy and 20 with superimposed preeclampsia) and 127 normotensive, healthy pregnant women were included in the study. Serum Hsp70 concentration was measured using enzyme-linked immunosorbent assay. The serum Hsp70 concentration was significantly higher in patients with transient hypertension of pregnancy, in preeclamptic patients and in patients with superimposed preeclampsia than in the control group (median (25-75 percentile): 0.66 (0.52-0.84), 0.55 (0.42-0.80), 0.61 (0.42-0.91) ng/ml vs 0.31 (0.27-0.39) ng/ml, respectively; P<0.001). Multivariate logistic regression analysis showed independent association of elevated serum Hsp70 level with transient hypertension of pregnancy, preeclampsia and superimposed preeclampsia. The difference in serum Hsp70 concentration between preeclamptic patients and the control group was statistically significant in each gestational age category. In the groups of preeclamptic and superimposed preeclamptic patients, there was no significant difference in serum Hsp70 concentration between mild and severe preeclamptic patients, between patients with late and early onset of the disease, as well as between preeclamptic patients without and with foetal growth restriction. In conclusion, serum Hsp70 concentration is elevated in transient hypertension of pregnancy, in preeclampsia and in superimposed preeclampsia. Circulating Hsp70 may not only be a marker for these conditions, but might also play a role in their pathogenesis. However, further studies are needed to explore its role in the pathogenesis of hypertensive disorders of pregnancy.