Phenotypic and metabolic characteristics of monocytes and granulocytes in preeclampsia

OBJECTIVE: The maternal syndrome of preeclampsia has recently been attributed to a systemic intravascular inflammatory response and endothelial cell activation and dysfunction. This novel hypothesis has considerable clinical and biological implications. This study was designed to determine whether women with preeclampsia have evidence of intravascular inflammation by examination of the phenotypic and metabolic activity of granulocytes and monocytes. STUDY DESIGN: A cross-sectional study was performed that included patients with preeclampsia (n = 31) and normal pregnancies (n = 58) matched for gestational age at blood draw. Intravascular inflammation was studied with use of flow cytometry. Peripheral venous blood was assayed to determine granulocyte and monocyte phenotype with the use of monoclonal antibodies for selective cluster differentiation (CD) antigens. The panel of antibodies included CD11b, CD14, CD16, CD18, CD49d, CD62L, CD64, CD66b, and HLA-DR. The quantity of basal intracellular reactive oxygen species and oxidative burst was assessed. Results were reported as mean channel brightness or intensity of detected fluorescence. Analysis was conducted with nonparametric statistics. A P value <.01 was considered to be significant. RESULTS: Preeclampsia was associated with a significant increase in mean channel brightness for CD11b on granulocytes and monocytes but lower mean channel brightness for CD62L on granulocytes than those from women with normal pregnancy (P <.01 for each). Basal intracellular reactive oxygen species were increased in monocytes but not in granulocytes. The oxidative burst was higher in both cell types. CONCLUSION: Preeclampsia is associated with phenotypic and metabolic changes in granulocytes and monocytes.     

Phenotypic and metabolic characteristics of maternal monocytes and granulocytes in preterm labor with intact membranes.

OBJECTIVE: Experimental and clinical studies support a role for the fetus in the control of the onset of labor. Fetal systemic inflammation, but not a maternal inflammatory response, has been linked to the onset of preterm labor and delivery on the basis of the determination of inflammatory cytokines in fetal and maternal blood. We propose that parturition requires fetomaternal cooperation and that inflammation is an integral part of the parturitional process. This study used flow cytometry, a sensitive technique for the detection of intravascular inflammation, to assess whether maternal inflammation is present in preterm labor. STUDY DESIGN: A prospective cross-sectional study was performed including patients with preterm labor (n = 55) and women with normal pregnancy (n = 50). Intravascular inflammation was studied by using flow cytometry. Maternal blood was assayed to determine granulocyte and monocyte phenotype by using monoclonal antibodies, which included the following cluster of differentiation (CD) markers: CD11b, CD14, CD15, CD16, CD18, CD49d, CD62L, CD64, CD66b, and HLA-DR. Oxidative burst and generation of basal intracellular oxygen radical species were assessed. Statistical analysis was conducted with the use of nonparametric methods. A P value of <.01 was considered statistically significant. RESULTS: Preterm labor was associated with a significant increase in the median mean channel brightness of CD11b, CD15, and CD66b on granulocytes and median mean channel brightness of CD11b and CD15 on monocytes. The ratio of oxidative burst over basal intracellular oxygen radical species in both granulocytes and monocytes was increased in preterm labor (P <. 01). CONCLUSION: Preterm labor with intact membranes is associated with phenotypic and metabolic changes of maternal granulocytes and monocytes.     

Leukocytes of pregnant women with small-for-gestational age neonates have a different phenotypic and metabolic activity from those of women with preeclampsia

Objective.?Preeclampsia and pregnancies complicated by small-for-gestational age (SGA) neonates share several underlying mechanisms of disease. However, while an exaggerated systemic maternal inflammatory response is regarded as one of the hallmarks of the pathogenesis of preeclampsia, the presence of a similar systemic intra-vascular inflammation in mothers of SGA neonates without hypertension is controversial. The aim of this study was to determine phenotypic and metabolic changes in granulocytes and monocytes of women who develop preeclampsia and those who deliver an SGA neonate, compared to normal pregnant women.

Methods.?This cross-sectional study included patients with a normal pregnancy (n?=?33), preeclampsia (n?=?33), and an SGA without preeclampsia (n?=?33), matched for gestational age at blood sample collection. Granulocyte and monocyte phenotypes were determined by flow cytometry, using monoclonal antibodies against selective cluster of differentiation (CD) antigens. The panel of antibodies included the following: CD11b, CD14, CD16, CD18, CD49d, CD62L, CD64, CD66b, and HLA-DR. Intracellular reactive oxygen species (iROS) were assessed at the basal state and after stimulation (oxidative burst). Results were reported as mean channel brightness (MCB) or intensity of detected fluorescence. Analysis was conducted with non-parametric statistics. A p-value?<?0.01 was considered statistically significant.

Results.?(1) Women who delivered an SGA neonate had a higher MCB of CD11b in granulocytes and monocytes than those with a normal pregnancy (p?<?0.001 for both); (2) patients with preeclampsia had a lower median MCB of CD62L in granulocytes (p?=?0.006) and a higher median basal iROS and oxidative burst in monocytes than women with an SGA neonate (p?=?0.003 and p?=?0.002, respectively).

Conclusion.?Pregnancies complicated by the delivery of an SGA neonate are characterized by a higher activation of maternal peripheral leukocytes than in normal pregnancies, but lower than in pregnancies complicated by preeclampsia.     

Neutrophil function in women with pre-eclampsia.

OBJECTIVE: To investigate the function of neutrophils in normal pregnancy and pre-eclampsia. DESIGN: Baseline levels and activated responses of peripheral blood neutrophils were measured in response to the physiological agonists, n-formyl-met-leu-phe (fMLP) and zymosan activated serum. SAMPLE: Neutrophils of 16 pre-eclamptic, 17 normal pregnant (third trimester) and 15 nonpregnant age-matched control women were calculated. SETTING: Antenatal Clinic, City Hospital, Nottingham. METHODS: Neutrophil superoxide anion production was determined by lucigenin-enhanced chemiluminescence; the release of secondary granule lactoferrin by ELISA; and the expression of cell surface adhesion molecules CD11b, CD18 and L-selectin (CD62L) by flow cytometric analysis. RESULTS: Superoxide anion generation was reduced in the pregnant group compared with nonpregnant controls [fMLP by 51% (P = 0.03) and zymosan activated serum by 56% (P = 0.01)] but pre-eclamptic measurements did not show a similar reduction. There were no differences between the three study groups in the plasma levels of lactoferrin, or in the stimulated expression and release of CD11b and CD18, or lactoferrin and L-selectin. The base-line levels for the production of superoxide anions; the expression of CD11b or CD18 or L-selectin; and the release of lactoferrin showed no significant differences. CONCLUSIONS: Circulating neutrophils in pregnancy and pre-eclampsia are neither activated nor primed in vivo, however the release of reactive oxygen species is diminished in normal pregnancy. In comparison, an elevation of reactive oxygen generation in pre-eclampsia may highlight a role for neutrophils in the oxidative stress and pathophysiology of this disease.     

The effect of healthy pregnant plasma and preeclamptic plasma on the phagocytosis index of neutrophil granulocytes and monocytes of nonpregnant women

Objective: Phagocyte function of neutrophil granulocytes and monocytes is decreased in healthy pregnancy and further decreased in preeclampsia. The cause of the declined function is unknown. Decreased phagocyte function can lead to the higher infection rate in healthy pregnancy and may also be responsible for the increased susceptibility to infections and high trophoblast concentration in preeclampsia. The aim of this study is to examine the phagocyte function of neutrophil granulocytes and monocytes. Methods: Monocytes and neutrophil granulocytes were separated from the peripheral circulation of six nonpregnant patients and incubated in plasma samples from six healthy pregnant, six preeclamptic pregnant, and six nonpregnant patients. The cells were marked and evaluated for the phagocytosis index with immunofluorescent microscope after phagocyting the zymosan molecules. Results: Phagocyte function of neutrophils as well as monocytes from nonpregnant patients were decreased significantly when the cells were incubated in plasma samples from healthy pregnant patients, and further decreased when incubated in plasma samples from preeclamptic pregnant women. Conclusion: The decreased phagocyte function of neutrophil granulocytes and that of monocytes in healthy pregnancy and the further decreased phagocyte function in preeclampsia is caused by factor(s) in the maternal circulation.     

Neutrophil function in women with pre-eclampsia

Objective To investigate the function of neutrophils in normal pregnancy and pre-eclampsia.
Design Baseline levels and activated responses of peripheral blood neutrophils were measured in response to the physiological agonists, n-formyl-met-leu-phe (fMLP) and zymosan activated serum.
Sample Neutrophils of 16 pre-eclamptic, 17 normal pregnant (third trimester) and 15 nonpregnant age-matched control women were calculated.
Setting Antenatal Clinic, City Hospital, Nottingham.
Methods Neutrophil superoxide anion production was determined by lucigenin-enhanced chemilumi-nescence; the release of secondary granule lactoferrin by ELISA; and the expression of cell surface adhesion molecules CD1 1b, CD 18 and L-selectin (CD62L) by flow cytometric analysis.
Results Superoxide anion generation was reduced in the pregnant group compared with nonpregnant controls [fMLP by 51 % (   P = 0.03  ) and zymosan activated serum by 56% (   P = > 0.01  )] but pre-eclamptic measurements did not show a similar reduction. There were no differences between the three study groups in the plasma levels of lactofenin, or in the stimulated expression and release of CD1 Ib and CD18, or lactoferrin and L-selectin. The base-line levels for the production of superoxide anions; the expression of CD11b or CD 18 or L-selectin; and the release of lactofemn showed no significant differences.
Conclusions Circulating neutrophils in pregnancy and pre-eclampsia are neither activated nor primed in vivo , however the release of reactive oxygen species is diminished in normal pregnancy. In comparison, an elevation of reactive oxygen generation in pre-eclampsia may highlight a role for neutrophils in the oxidative stress and pathophysiology of this disease.     

Maternal peripheral blood leukocytes in normal and pre-eclamptic pregnancies.

OBJECTIVE: To analyse activation of maternal peripheral blood leukocytes by flow cytometric measurements of intracellular free-ionised calcium of lymphocytes, granulocytes and monocytes, separately. DESIGN: Case-control study. SETTING: High risk pregnancy service in a regional centre. MATERIAL: Samples from 10 women with pre-eclampsia, 10 appropriately-matched women with normal pregnancy, nine multigravid normal women at mid-gestation selected as being least likely to demonstrate any tendencies towards pre-eclampsia, and 11 healthy nonpregnant women of reproductive age were studied. METHODS: Using flow cytometry, intracellular free ionised calcium ([Ca2+]i) was estimated by loading the cells with Fluo-3 and measuring the changes in fluorescence intensity induced by free ionised calcium. After the basal levels were measured, the response of phagocytes to stimulation with n-formylmethionyl-leucyl-phenylalanine (fMLP) was determined. MAIN OUTCOME MEASURES: Basal [Ca2+]i of peripheral blood leukocytes. RESULTS: Median basal [Ca2+]i was significantly increased in all three subsets of leukocytes--lymphocytes, granulocytes and monocytes in pre-eclampsia--compared with the three control groups. Samples from both groups of women with normal pregnancy did not differ from those from nonpregnant women. The peak responses of monocytes to stimulation with 10 nmol fMLP were greater in samples from pre-eclamptic women, giving evidence of priming. CONCLUSIONS: Peripheral blood leukocytes are activated in pre-eclampsia in terms of basal changes in the intracellular second messenger--free ionised calcium. Peripheral blood monocytes are primed to give greater responses after stimulation with fMLP.     

Maternal peripheral blood leukocytes in normal and pre-eclamptic pregnancies

Objective To analyse activation of maternal peripheral blood leukocytes by flow cytometric measurements of intracellular free-ionised calcium of lymphocytes, granulocytes, and monocytes, separately.
Design Case-control study.
Setting High risk pregnancy service in a regional centre.
Material Samples from 10 women with Pre-eclampsia, 10 appropriately-matched women with normal pregnancy, nine multigravid normal women at mid-gestation selected as being least likely to demonstrate any tendencies towards Pre-eclampsia, and 11 healthy nonpregnant women of reproductive age were studied.
Methods Using flow cytometry, intracellular free ionised calcium ([Ca²⁺]_i) was estimated by loading the cells with Fluo-3 and measuring the changes in fluorescence intensity induced by free ionised calcium. After the basal levels were measured, the response of phagocytes to stimulation with n-formylmethionyl-leucyl-phenylalanine (fMLP) was determined.
Main outcome measures Basal [Ca²⁺]_i of peripheral blood leukocytes.
Results Median basal [Ca²⁺]_i was significantly increased in all three subsets of leukocytes—lymphocytes, granulocytes and monocytes in Pre-eclampsia—compared with the three control groups. Samples from both groups of women with normal pregnancy did not differ from those from nonpregnant women. The peak responses of monocytes to stimulation with 10 nmol fMLP were greater in samples from pre-eclamptic women, giving evidence of priming.
Conclusions Peripheral blood leukocytes are activated in Pre-eclampsia in terms of basal changes in the intracellular second messenger—free ionised calcium. Peripheral blood monocytes are primed to give greater responses after stimulation with fMLP.     

Granulocyte superoxide anion production and regulation by plasma factors in normal and preeclamptic pregnancy

Data on the respiratory burst activity of granulocytes from healthy and preeclamptic women have remained contradictory. To investigate the role of reactive oxygen species in the etiology of preeclampsia we measured superoxide anion generation by granulocytes from non-pregnant, healthy, and preeclamptic women. We also examined the reciprocal effects of heat-inactivated and non-inactivated plasma on superoxide production. Superoxide generation was measured by ferricytochrome-c reduction. Superoxide production induced by either phorbol-12,13-dibutirate or N-formyl-methionyl-leucyl-phenylalanine was significantly decreased in granulocytes from normal pregnant women compared with non-pregnant and preeclamptic women. The phorbol-12,13-dibutirate-induced superoxide generation by granulocytes from non-pregnant and preeclamptic women was significantly inhibited by plasma from healthy pregnant women. The N-formyl-methionyl-leucyl-phenylalanine-stimulated superoxide production by granulocytes from non-pregnant and preeclamptic women was suppressed only by non-inactivated plasma, not heat-inactivated plasma from healthy pregnant women. Plasma from preeclamptic women did not influence the phorbol-12,13-dibutirate- and N-formyl-methionyl-leucyl-phenylalanine-induced superoxide production by control granulocytes. The phorbol-12,13-dibutirate-induced superoxide generation by granulocytes from healthy pregnant women was significantly increased by the effect of plasma from non-pregnant and preeclamptic women, but when stimulating with N-formyl-methionyl-leucyl-phenylalanine only non-inactivated plasma caused the same enhancement. These data indicate that reduced superoxide generation in normal pregnancy may be caused by maternal immunosuppressive factors present in plasma. The failure to reduce superoxide production in preeclampsia may be partly responsible for the endothelial dysfunction characteristic of that condition.     

Activation of peripheral leukocytes in rat pregnancy and experimental preeclampsia

OBJECTIVE: The aim of this study was to search for activation markers of peripheral leukocytes in experimental preeclampsia in the rat. STUDY DESIGN: Experimental preeclampsia was induced in 14-day-pregnant rats by infusion of endotoxin (1.0 microg/kg body weight). For comparison, rats with normal pregnancies that were infused with sodium chloride solution and cyclic rats that were infused with either endotoxin or sodium chloride solution were used. At various points before and after the infusion, blood samples were withdrawn and analyzed by means of whole-blood flow cytometry to evaluate expression of inflammation-associated adhesion molecules (CD11b, CD11a, CD49d, and CD62L) and CD14 on the leukocytes. RESULTS: Normal pregnancy was associated with increased CD11b (granulocytes and monocytes), CD11a (monocytes and lymphocytes), and CD49d (granulocytes, monocytes, and lymphocytes) expression. In addition to these changes found in normal pregnancy, reduced CD62L and increased CD11a and CD49d expression was found on granulocytes after endotoxin treatment of pregnant rats. No effect of endotoxin was observed in cyclic rats. CONCLUSION: Leukocytes of rats with experimental preeclampsia and, to a lesser extent, those of rats with normal pregnancies had an activated phenotype. These results are consistent with our previous findings in human subjects and suggest that (experimental) preeclampsia results from a generalized inflammatory response.     

Differences in innate immunologic response to group B streptococcus between colonized and noncolonized women

OBJECTIVE: To evaluate the functional capacity of granulocytes and monocytes from pregnant and nonpregnant women in relation to group B streptococcus (GBS) colonization status. METHODS: Engulfment of fluorescent GBS by peripheral blood phagocytes from GBS-colonized and noncolonized women was measured by flow cytometry. Intracellular superoxiode generated in response to GBS challenge to monocytes and granulocytes enriched from peripheral blood of these women was also measured by flow cytometry, and extracellular superoxide was determined by colorimetric assay. RESULTS: Monocytes and granulocytes from pregnant, GBS-colonized women engulfed significantly greater numbers of GBS than phagocytes from pregnant, noncolonized women. No difference in intracellular superoxide production was detected between any of the groups of women; however, monocytes from pregnant, colonized women released significantly more superoxide into the extracellular milieu than did granulocytes from the same women. No differences in extracellular release of superoxide were observed among noncolonized women whether they were pregnant or not. CONCLUSIONS: Monocytes from pregnant, colonized women engulf more GBS and release more of the superoxide into the extracellular environment, where it is unlikely to be an effective defense mechanism against intracellular bacteria. This suggests that components of the innate immune system that should serve in a protective role may function suboptimally, thereby contributing to the colonization process by GBS.     

Subpopulations of t lymphocytes and lymphocyte proliferative activity in normal pregnancy]

T lymphocyte subpopulations (CD3+, CD4+, CD8+) and the lymphocyte proliferative responses to mitogens (PHA, Con A, PWM), in the environment of fetal calf serum (FCS) were examined in 16 normal primigravidas in the third trimester of pregnancy and in 15 healthy nonpregnant women. In normal pregnant women significantly lower absolute and percentage numbers of CD3+ and CD4+ T cells and almost twice reduced CD4+/CD8+ ratio were found, in comparison with nonpregnant subjects. Despite the shifts among particular T lymphocyte subsets in the peripheral blood, non disorders were found in the lymphocyte proliferative responses to mitogens in normal pregnancy.     

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.     

Serum adenosine deaminase activity and its isoenzyme pattern in women with normal pregnancies

Adenosine deaminase (ADA) is a purine enzyme which is essential for the proliferation, maturation and function of lymphoid cells, and congenital deficiency of this enzyme is associated with severe combined immunodeficiency disease. The activity of ADA has changed in diseases characterized by the alteration of cell-mediated immunity such as rheumatoid arthritis, systemic lupus erythematosus and tuberculosis, so ADA has been considered as a nonspecific marker of cell-mediated immunity. In this study we examined changes in serum total ADA activity and the patterns of two ADA isoenzymes, ADA1 and ADA2 in normal pregnant women, and evaluated the possible role of the alteration of cell-mediated immunity during normal pregnancy as causes of changes in ADA activity. We measured serum activities of total ADA, ADA1 and ADA2 in normal pregnant women in the third trimester (n=24) and age-matched healthy nonpregnant women (n=24). Peripheral blood lymphocytes and monocytes were also measured. In normal pregnant women, serum total ADA activity averaged 10.5 +/- 0.5 U/L, which was significantly lower than in nonpregnant women (14.0 +/- 0.5 U/L ) (p<0.05), and mean serum ADA2 activity also significantly reduced that of nonpregnant women (p<0.05). There was no significant difference in ADA1 activity in normal pregnant and nonpregnant women. The decrease in total ADA activity was accompanied by the decrease in lymphocyte count. These results suggest that reduced serum total ADA activity reflects decrease in ADA2 activity, and which may be in part associated with depressed cell-mediated immunity during normal pregnancy.     

Increased breath markers of oxidative stress in normal pregnancy and in preeclampsia

OBJECTIVES: The purpose of this study was to compare the intensity of oxidative stress in normal pregnancy, preeclampsia, and nonpregnant women using a breath test. STUDY DESIGN: We studied primiparous women in third trimester pregnancy (38 uncomplicated, 26 with preeclampsia) and 60 nonpregnant control subjects. Volatile organic compounds (VOCs) in alveolar breath were analyzed by gas chromatography/mass spectroscopy to construct the breath methylated alkane contour (BMAC), a 3-dimensional display of abundance of C4-C20 alkanes and monomethylated alkanes. RESULTS: The mean volume under curve (VUC) of the BMAC was significantly higher in preeclampsia patients than in normal pregnant women (P < .003) and nonpregnant control subjects (P < .005). A predictive model employing 5 VOCs distinguished preeclampsia from uncomplicated pregnancy (sensitivity = 92.3%, specificity = 89.7%; cross-validated sensitivity = 88.5%, specificity = 79.3%). CONCLUSION: A breath test significantly demonstrated greater oxidative stress in women with preeclampsia than in uncomplicated pregnancy and nonpregnant control subjects. The breath test accurately identified women with established preeclampsia, but further studies are required to determine if this test can predict the onset of disease.     

Oxidant release is dramatically increased by elevated glucose concentrations in neutrophils from pregnant women.

OBJECTIVE: To evaluate the mechanism of oxidative stress at glucose levels accompanying diabetic pregnancy. Specifically, we hypothesize that elevated glucose overwhelms hexose monophosphate shunt (HMS) down-regulation observed during pregnancy. METHODS: Peripheral blood cells from normal healthy pregnant women were exposed to heightened glucose levels to provide an in vitro model of the effects of diabetic pregnancy. Changes in NAD(P)H, reactive oxygen species (ROS) and nitric oxide (NO) production were evaluated in single cells. RESULTS: Altered metabolic dynamics, as judged by NAD(P)H autofluorescence of neutrophils from both pregnant and non-pregnant women, were observed during incubation with 14 mM glucose, a pathophysiologic level. In parallel, increased production of ROS and NO was observed. The ROS and NO levels attained in cells from pregnant women were greater than those observed in cells from non-pregnant women. Inhibitors of the HMS and NAD(P)H oxidase blocked these effects. These metabolic and oxidant changes required approximately one minute, suggesting that transient glucose spikes during pregnancy could trigger this response. CONCLUSIONS: Elevated glucose levels enhance HMS activity and oxidant production in cells from pregnant women. This mechanism may be generally applicable in understanding the role of diabetes in materno-fetal health.     

Nonspecific immunity in pregnancy: monocyte surface Fcγ receptor expression and function

The state of pregnancy is an immunological enigma during which the body must prevent rejection of the antigenically foreign fetus while at the same time maintain sufficient maternal host defense mechanisms to combat infection. Although most studies on the immunology of pregnancy focus on immune suppression, several studies have shown an increase in nonspecific host defense, which is postulated to be a compensatory mechanism for decreased specific immunity during pregnancy. Studies in this laboratory have shown that monocyte surface FcγRI (CD64) and FcγRII (CD32) expression progressively increase throughout pregnancy, while surface MHC class II expression remains unchanged. Functional studies revealed that the number of phagocytic monocytes which could be isolated from pregnant women was increased. These cells exhibited an increased capacity to ingest IgG-opsonized human erythrocytes. This study shows for the first time that monocyte surface FcγR expression and FcγR-mediated functions are increased during pregnancy. These results support the hypothesis that nonspecific immunity as represented by FcγR expression and function is increased during pregnancy.     

Serum lipid peroxidation and antioxidant potential levels in hyperemesis gravidarum

Hyperemesis gravidarum (HEG) is a severe form of nausea and vomiting with unknown etiology. Recent studies have suggested that there might be an etiologic role for HELICOBACTER PYLORI (HP) in HEG. HP is a Gram-negative bacterium that colonizes the gastric mucosa, increases the generation of reactive oxygen species (ROS), and decreases plasma antioxidants such as ascorbic acid. The aim of this study was to investigate plasma ROS activities and antioxidant status, and their relationship with HP infection, in HEG womens. Twenty-five HEG women, 20 gestational age- and gravida-matched healthy pregnant controls, and 15 nonpregnant women were examined for specific immunoglobulin G (IgG) against HP, serum malondialdehyde (MDA) as a measure of lipid peroxidation, and activity of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CT). Compared with controls, the seropositivity of HP for IgG was significantly higher ( P < 0.01) in pregnancies with HEG. When three study groups (including both HP + and HP- patients) were compared with each other with respect to the study parameters, MDA concentration increased significantly in pregnant women when compared with control group (nonpregnant women; P < 0.01), as well as in HEG compared with healthy pregnant women ( P < 0.01). The SOD, GSH-Px, and CT activity were increased in the group of normal pregnant women versus the nonpregnant group ( P < 0.01), but decreased significantly in the group of HEG pregnant women ( P < 0.01). Serum MDA levels and SOD, GSH-Px, and CT activity were not affected by the seropositivity of HP for IgG in either group ( P > 0.05). The results of the present study suggest that HEG is an oxidative stress condition, as reflected by the increased ROS activity and decreased antioxidant status, regardless of HP infection.     

Different levels of platelet activation in preeclamptic, normotensive pregnant, and nonpregnant women

OBJECTIVE: The aims of our study were to determine the basal platelet activation state in women with preeclampsia compared with normotensive pregnant women and nonpregnant women and to investigate the platelet reactivity on in vitro stimulation with adenosine diphosphate or thrombin receptor activation peptide. STUDY DESIGN: Platelet expression of CD61 (fibrinogen receptor), CD42a (von Willebrand factor receptor), CD62P (P-selectin), CD63 (Glycoprotein 53), and PAC-1 binding (activated fibrinogen receptor) were determined in 20 pairs of women with preeclampsia/normotensive pregnant women and in 12 nonpregnant women, with the use of flow cytometry. RESULTS: Basal platelet expression of CD61, CD42a and CD62P, and adenosine diphosphate-stimulated CD62P expression were increased in women with preeclampsia compared with normotensive pregnant women. Platelets from women with preeclampsia and normotensive pregnant women differed from platelets from nonpregnant women by expressing higher basal CD63 levels and being more responsive to in vitro agonist stimulation, which was demonstrated by increased expression of CD61, CD62P, and CD63. CONCLUSION: This study supports the notion that platelets are important in the pathophysiologic condition of preeclampsia.     

Expression of intercellular adhesion molecule-1 (ICAM-1) on the surface of peripheral blood lymphocytes of pregnant women with pregnancy-induced hypertension

OBJECTIVES: Intercellular adhesion molecule-1 (ICAM-1, CD54) is involved in process of leukocytes adhesion to endothelium as well as in their migration to surrounding tissues. There is much evidence that pregnancy-induced hypertension (PIH) presents a state of endothelial destruction mediated partially by increased ICAM-1 expression on endothelial cells and neutrofils. DESIGN: The aim of this study was to evaluate the expression of ICAM-1 (CD54) molecule on the peripheral blood lymphocytes of pregnant women with PIH studied "in vitro". MATERIALS AND METHODS: Preeclampsia (PE) and transient hypertension (TH) were defined according to USA National Health Institute criteria. The study group consisted of 16 women with preeclampsia (PE), 12 women with transient hypertension (TH) and 9 women with physiological pregnancy. The group of 8 nonpregnant women served as controls. Exclusion criteria were: uterine contractions, infection and steroid therapy before blood sampling. Peripheral blood was obtained by venipuncture. Lymphocytes were isolated and cultured by using standard procedures. Mitogenic doses of phytohaemaglutynin (PHA) were added to each culture. Immunofluorescent marking techniques with anty-CD54 one-step monoclonal antibodies were performed. Analysis was made with FACSCalibur flow-cytometer with 488 nm argon laser using CellQuest programme. The results were described as the percentage of CD54+ lymphocytes and MFI index corresponding density of CD54 molecules on the lymphocyte surface. Statistical analysis was performed using t-Student and U-Mann-Whitney tests. The work was sponsored by KBN 4 P05E 118,15 grant. RESULTS: The percentage of CD54+ lymphocytes in physiological pregnancy compared to nonpregnant women did not differ significantly (56.9 +/- 20.8% vs. 57.2 +/- 14.0%, p = 0.97). The MFI value was increased in pregnant women but in comparison with nonpregnant women did not reach statistical significance (34.7 +/- 35.7 vs. 17.8 +/- 4.3, p = 0.20). The percentage of CD54+ lymphocytes in TH group compared to normal pregnant women did not differ significantly (52.2 +/- 18.6% vs. 56.9 +/- 20.8%, p = 0.58) but MFI value was significantly increased (100.6 +/- 81.5 vs. 34.7 +/- 35.7). In PE group compared to normal pregnant women the percentage of CD54+ lymphocytes as well as MFI value were significantly increased (CD54+: 70.8 +/- 12.9% vs. 56.9 +/- 20.8%, p < 0.05; MFI: 170.8 +/- 91.7 vs. 34.7 +/- 35.7, p < 0.0005). CONCLUSIONS: 1/expression of ICAM-1 molecule on peripheral blood lymphocytes studied "in vitro" during normal pregnancy is not different in comparison to the nonpregnant state, but 2/ in pregnancy complicated with PIH is significantly increased, especially in PE, 3/described changes are a sign of the lymphocyte activation and may be responsible for endothelial destruction observed in PIH.