The anti-tumor immune responses induced by a fusion protein of ovarian carcinoma anti-idiotypic antibody 6B11ScFv and murine GM-CSF in BALB/c mice

Ovarian carcinoma anti-idiotypic antibody 6B11 was murine derived; we previously have cloned 6B11 single-chain Fv antibody (6B11ScFv) and constructed the 6B11ScFv/human granulocyte-macrophage colony stimulating factor (GM-CSF) fusion protein (designated as 6B11GM) to enhance the immunogenecity of the single-chain Ab(2). Because of the difference in species specificity between human GM-CSF and murine GM-CSF, there is no immune competent animal model on which the effect and metabolism of 6B11GM as a vaccine could be observed. In this study, 6B11mGM fusion gene was constructed by the fusing murine GM-CSF cDNA gene with 6B11ScFv. The fusion gene was cloned and expressed. The product of this gene is a fusion protein. It could specifically interact with the primary anti-ovarian carcinoma monoclonal antibody (COC166-9) and rat anti-mouse GM-CSF monoclonal antibody, respectively, and stimulate the growth of NFS-60 cells (a murine GM-CSF-dependent cell line). The specific anti-tumor immune response could be induced in BALB/c mice after immunized with anti-idiotypic fusion protein instead of ovarian carcinoma antigen without carrier proteins and adjuvant. Ab(3) could be detected in the sera of immunized mice with 6B11mGM by enzyme-linked immunoadsorbent assay test. Moreover, the fusion protein stimulated proliferation of CD4+ T cell from the spleen of BALB/c mice and proliferation of CD8+ T cell to a lesser degree. Therefore, 6B11mGM probably induces both humoral and cellular immunity against ovarian carcinoma in vivo.     

Alpha-fetoprotein and hematopoietic growth factors in amniotic fluid

OBJECTIVE: To determine whether a relationship exists between alpha-fetoprotein (AFP) and hematopoietic growth factors in amniotic fluid. METHODS: Forty-one women at 15 weeks' gestation were included in the study. Gestational age was assessed by obtaining a reliable menstrual history and scanning. Amniocentesis was performed, and each woman subsequently delivered anatomically and chromosomally normal infants. The level of AFP was determined using a standard automated procedure. The concentrations of stem cell factor, interleukin 3, interleukin 6, erythropoietin, and granulocyte colony-stimulating factor (G-CSF) were measured using a commercially available immunoassay. The relationships between AFP and the studied cytokines were evaluated using the Pearson linear correlation test. Significant correlations were studied further by linear and nonlinear regression to obtain the best predictive model. RESULTS: There was a significant correlation between AFP and stem cell factor (r =.47, P =.002). No significant correlations between AFP and the rest of the studied cytokines were found (r = -.07, r =.02, r = -.02, and r = -.11 for erythropoietin, G-CSF, interleukin 3, and interleukin 6, respectively). CONCLUSION: Alpha-fetoprotein is significantly correlated with stem cell factor in early pregnancy and might play a role in fetal hematopoiesis.     

The Presence of Cytokines and Growth Factors in Hydrosalpingeal Fluid

PURPOSE: To examine the presence of cytokines and growth factors in hydrosalpingeal fluid. METHODS: Eighteen hydrosalpingeal fluids were compared with 15 follicular fluids and serum samples regarding the presence of interleukin-8 (IL-8), IL-12, IL-1alpha, epidermal growth factor (EGF), granulocyte macrophage colony stimulating factor (GM-CSF), leukemia inhibitory factor (LIF), tumor necrosis factor-alpha (TNFalpha), interferon-gamma (IFNgamma), and transforming growth factor-beta2 (TGFbeta2). RESULTS: IL-8 and EGF were detected in all the hydrosalpinx samples. IL-8, IL-12, IL-1alpha, TNFalpha, TGFbeta2, GM-CSF, and LIF were detected to a significantly larger extent in hydrosalpingeal than follicular fluids (p < 0.01). The same cytokines, with the exception of IL-8, TGFbeta2, and LIF, were also more frequently present in comparison with serum. CONCLUSION: The abundant presence of cytokines in hydrosalpingeal fluid suggests an increased expression from the tubal epithelium. Whether high concentrations have a negative influence on embryo development and implantation needs further investigation.     

Cytokine profile as diagnostic and prognostic factor in neonatal sepsis

Antecedents: The serum levels of some cytokines can be useful in the diagnosis of neonatal sepsis; the prognostic value of a cytokine profile has not, to our knowledge, been explored in this disease.

Objective: The objective of this study is to evaluate the diagnostic value of the serum levels of cytokines IL-1, -2, -4, -5, -6, -7, -8, -10, -12, -13, and -17, TNF, IFNγ, G-CSF, GM-CSF, MCP1, and MIP1β in neonates with high risk of developing sepsis.

Methods: Sepsis was evaluated in 96 high-risk neonates. We assessed cytokine levels on hospital admission and during or not during sepsis.

Results: Fifty (52%) presented sepsis (26 early and 24 late). Sepsis was associated with high levels of IL-6, IL-10, G-CSF, and MCP1 and low levels of IFNγ, early sepsis with high levels of IL-6 and G-CSF, severe sepsis with high levels of IL-6 and IL-10, while deaths or sequelae was associated with low levels of IL-4, IL-12, IFNγ, and high levels of GM-CSF. IL-6 values of ≥40.1?pg/mL were associated with the development of any type of sepsis (relative risk [RR]: 1.70; 95% confidence interval [95% CI]: 1.18–2.24; p?=?.01), while IL-6 values of ≥44.9?pg/mL were associated with early sepsis (RR: 1.29; 95% CI: 1.29–4.56; p?=?.01).

Conclusion: In neonates with high risk for the development of sepsis, there is an association between levels of IL-6, IL-10, and G-SCF and the disease development/outcome.     

Peritoneal fluid cytokine and eicosanoid levels and their relation to the incidence of peritoneal adhesion.

OBJECTIVE: To determine the peritoneal fluid content of several cytokines and eicosanoids with inflammatory, anti-inflammatory, anti-fibrotic, and fibrotic activities, and to assess the relationship of these levels with the incidence of peritoneal adhesions. METHODS: Peritoneal fluids were collected from 30 subjects with adhesions (n = 22) or with normal pelvic anatomy (n = 8), and the level of interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha), interleukin-10 (IL-10), interferon gamma (IFN-gamma), transforming growth factor beta-1 (TGF-beta 1), granulocyte macrophage-colony stimulating factor (GM-CSF), prostaglandin E2 (PGE2), leukotriene B4 (LTB4), and thromboxane B2 (TXB2) were determined by enzyme-linked immunosorbent assay (ELISA) and radioreceptor assay. RESULTS: The peritoneal fluid content of these factors varied considerably, with low levels of IL-1 beta, TNF-alpha, IL-10, IFN-gamma, and GM-CSF. Only IFN-gamma levels were significantly lower in subjects with adhesions compared with the normal group (P < .05). The levels of total (latent + active) and active TGF-beta 1 were higher than those of other cytokines assayed and were significantly higher in subjects with adhesions compared with the normal group (P < .05). The peritoneal fluid content of PGE2, TXB2, and LTB4 was significantly higher than that of the cytokines and was higher, but not significantly so, in subjects with adhesions compared with normal subjects (P = .06). CONCLUSION: Although the effect of length of time since the adhesions were formed is not known, the results indicate that peritoneal fluid content of these cytokines and eicosanoids, with the exception of IFN-gamma and TGF-beta 1, does not correlate with the presence of peritoneal adhesions.     

Maternal plasma cytokines in early- and mid-gestation of normal human pregnancy and their association with maternal factors

Few studies have assessed longitudinal changes in circulating cytokine levels during normal pregnancy. We have examined the natural history of maternal plasma cytokines from early- to mid-pregnancy in a large, longitudinal cohort. Multiplex flow cytometry was used to measure interleukin (IL)-2, IL-6, IL-12, tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma and granulocyte-macrophage colony-stimulating factor (GM-CSF) in early- (median [IQR]: 8.5 weeks [7.1, 10.0]) and mid-pregnancy (25.0 [24.1, 26.1]) from 1274 Danish women delivering singleton term infants. GM-CSF decreased from early- to mid-pregnancy (median percent change [95% CI]: -51.3% [-59.1%, -41.8%]), while increases were observed in IL-6 (24.3% [4.6%, 43.9%]), IL-12 (21.3% [8.9%, 35.7%]) and IFN-gamma (131.7% [100.2%, 171.6%]); IL-2 (-2.8% [-11.5%, 0.0%]) and TNF-alpha (0% [-5.9%, 25.6%]) remained stable. Positive correlations were found between all cytokines, both in early- and mid-pregnancy (all p<0.001). Early- and mid-pregnancy levels were rank-correlated for IL-2, IL-12, TNF-alpha and GM-CSF, but not IL-6 and IFN-gamma; these correlations were generally weaker than correlations between different cytokines at a single time point in pregnancy. Women with a pre-pregnancy BMI <18.5 had reduced levels of IFN-gamma and GM-CSF compared to women in other BMI categories, while women aged >or=35 years had elevated IL-2, IL-6, TNF-alpha and IFN-gamma. Early-pregnancy levels of TNF-alpha were higher in women with a prior preterm delivery. Cytokine levels were not associated with gravidity. In conclusion, cytokines were detected in plasma during early- and mid-pregnancy, with IL-6, IL-12, IFN-gamma and GM-CSF concentrations varying over pregnancy. Concentrations may depend on BMI, maternal age and prior preterm delivery.     

State-of-the-Art Lectures

The pathophysiology of preeclampsia (PET) implicates an inflammatory dysfunction. This study profiled this host response by challenging whole blood with lipopolysaccharide. Multiplex immunoassays determined interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12(p70), IL-13, IL-17, granulocyte/granulocyte macrophage-colony stimulating factors (G-CSF/GM-SCF), interferon(IFN)-γ, monocyte chemotactic protein (MCP)-1, macrophage inflammatory protein-1β and tumor necrosis factor (TNF)-α levels. Secretory capacity was expressed in pg/million white cells or monocytes (±SEM). PET featured significantly higher IL-1β, IL-2, IL-10, IL-13, G-CSF, IFN-γ, MCP-1 and TNF-α monocyte secretory capacities (p < 0.05). The PET group exhibited an inflammatory hyper-responsiveness (p < 0.01) which was poorly described by the traditional Th1:Th2 dichotomy.     

Serum cytokines as biomarkers for nonsurgical prediction of endometriosis

OBJECTIVE: To test the ability of a group of serum cytokines, either individually or in combination, to serve as biomarkers for the nonsurgical diagnosis of endometriosis. STUDY DESIGN: Subjects were allocated to two groups according to their laparoscopic diagnosis. The first group consisted of patients with endometriosis and the second group was made up of infertile women with no pelvic pathology (controls). Blood samples were collected preoperatively and stored. Cytokines were measured in the serum of all participants using the Bio-Plex Protein Array System. Nonparametric statistics and the Mann-Whitney test were used to compare groups. Subjects were seen at the Gynecologic endoscopy unit. RESULTS: Three cytokines were significantly higher in the serum of subjects with endometriosis than in the control group: interleukin-6 (IL-6) [4.41 pg/ml (range: 1.47-15.01) versus 0.97 pg/ml (range: 0.29-2.98), respectively; p<0.001], monocyte chemotactic protein-1 (MCP-1) [37.91 pg/ml (range: 24.54-94.74) versus 22.13 pg/ml (range: 13.85-39.45), respectively; p<0.001], and interferon-gamma (INF-gamma) [19.01 pg/ml (range: 1.19-73.52) versus 0.30 pg/ml (range: 0.00-13.05), respectively; p<0.001]. There was no statistically significant difference between subjects with endometriosis and controls in the serum concentration of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), or granulocyte macrophage colony stimulating factor (GM-CSF). Interleukin-2 (IL-2), interleukin-8 (IL-8), and interleukin-15 (IL-15) were undetectable in the serum of both groups. None of the measured cytokines showed significant correlation with the cycle phase or stage of endometriosis. In a multivariate analysis, serum interleukin-6 provided a sensitivity of 71% and a specificity of 66% to discriminate between endometriosis patients and controls at a cutoff point of 1.9 pg/ml. Adding monocyte chemotactic protein-1 and interferon-gamma to interleukin-6 did not increase the discriminative ability over that achieved by measuring serum interleukin-6 alone. CONCLUSIONS: Serum of subjects with endometriosis contains significantly higher levels of interleukin-6, monocyte chemotactic protein-1, and interferon-gamma than control women. Serum interleukin-6 measurements discriminate between women with endometriosis and controls. Interleukin-6 provides a promising serum marker for the nonsurgical prediction of endometriosis.     

Host inflammatory response profiling in preeclampsia using an in vitro whole blood stimulation model.

The pathophysiology of preeclampsia (PET) implicates an inflammatory dysfunction. This study profiled this host response by challenging whole blood with lipopolysaccharide. Multiplex immunoassays determined interleukin (IL)-1beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12(p70), IL-13, IL-17, granulocyte/granulocyte macrophage-colony stimulating factors (G-CSF/GM-SCF), interferon(IFN)-gamma, monocyte chemotactic protein (MCP)-1, macrophage inflammatory protein-1beta and tumor necrosis factor (TNF)-alpha levels. Secretory capacity was expressed in pg/million white cells or monocytes (+/-SEM). PET featured significantly higher IL-1beta, IL-2, IL-10, IL-13, G-CSF, IFN-gamma, MCP-1 and TNF-alpha monocyte secretory capacities (p < 0.05). The PET group exhibited an inflammatory hyper-responsiveness (p < 0.01) which was poorly described by the traditional Th1:Th2 dichotomy.     

Gestational effects on host inflammatory response in normal and pre-eclamptic pregnancies

Objective

Pre-eclampsia (PET) remains a leading cause of maternal and neonatal morbidity and mortality. Although its pathophysiology involves an underlying inflammatory dysfunction, it is unclear how this may be affected by increasing gestational age, particularly in relation to the time of onset of disease. Murine studies have indicated that a progressive increase in serum inflammatory profile is a physiological feature of normal gestation. The present study aimed to investigate this phenomenon in women in relation to normal and pre-eclamptic pregnancies.

Study design

Control and PET groups (each n = 20) were divided into early and late pregnancy (before and after 34 weeks gestation, respectively). Whole blood was diluted 1:1 with RPMI 1640 medium with/without 1 μg/ml lipopolysaccharide at 37 °C for 24 h under a humidified 5% CO₂ atmosphere. Samples were collected at 0, 2, 6 and 24 h and analysed for interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, IL-17, granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon (IFN)-γ, monocyte chemotactic protein (MCP-1), macrophage inflammatory protein (MIP)-1β and tumour necrosis factor (TNF)-α by fluid-phase multiplex immunoassay.

Results

This study confirms that pregnancy features an increasing inflammatory response with advancing gestational age, which was seen in both control and PET pregnancies (P < 0.01).

Conclusions

This increase in inflammatory responsiveness with advancing gestation may provide an explanation for the incidence of late onset PET in the absence of placental pathology, as well as serving as a potential physiological priming mechanism geared towards increasing maternal sensitivity to the fetal triggers of labour.     

Granulocyte-macrophage colony stimulating factor (GM-CSF) and co-culture can affect post-thaw development and apoptosis in cryopreserved embryos

Purpose: The objective of this study was to evaluate the effects of growth factor supplementation and Vero cell co-culture on apoptosis and development of frozen thawed one-cell mouse embryos. Methods: The following treatment regimens were assessed: (a) control medium (b) Vero cell co-culture and (c) growth factor supplemented medium. The individual growth factors tested were: GM-CSF, IGF-I, IGF-II, TNF-α, FGF-4, LIF, TGF-α, TGF-β, IL-6, PDGF and EGF. Blastocyst development and differentiation were monitored. At termination of the experiments, overall blastomere number and apoptosis were assessed using the TUNEL assay. Results: No differences were observed in blastulation and hatching rates. ICM differentiation in thawed embryos was notably improved with either co-culture or growth factor supplementation. The only growth factor significantly modulating apoptosis in thawed embryos was granulocyte-macrophage colony stimulating factor (GM-CSF). GM-CSF enhanced continued cell survival and prevented apoptosis but did not influence overall cell number in developing blastocysts. Vero cell co-culture significantly increased cell number in blastocysts (124±42 vs 100±44 in control; P<0.05). Embryonic apoptosis was higher in the co-cultured embryos. The increased presence of apoptotic cells in blastocysts of high cell number may reflect the regulatory role of apoptosis in balancing ICM: TE ratios. Conclusion: These data indicate that culture conditions can modulate post-thaw embryonic development and apoptosis. Presented in part at: the 60th annual meeting of the American Society for Reproductive Medicine, 2004, October 16–20, Philadelphia, PA, USA.     

The effect of recombinant GM-CSF on IL-6 and TNF-alpha levels in epithelial ovarian cancer patients who received paclitaxel and cisplatinum: preliminary results

OBJECTIVE: We investigated the effects of GM-CSF factor on IL-6 and TNF-alpha levels prior to paclitaxel-cisplatinum combination chemotherapy for advanced epithelial ovarian cancer. MATERIALS AND METHODS: Twenty-three consecutive patients with FIGO (International Federation of Gynecology and Obstetrics) Stage III-IV epithelial ovarian cancer were enrolled in the study. Following cytoreductive surgery patients received 175 mg/m2 paclitaxel and 75 mg/m2 cisplatinum on the same day. These 23 patients also received RhuGM-CSF five days before at a dose of 5 microg/kg/day by subcutaneous injection for three days. IL-6 and TNF-alpha levels were measured before and 24 hours later following the last dose of RhuGM-CSF. RESULTS: White blood cell counts on the 10th day of the cycle were lower than preGM-CSF white blood cell counts and the difference was statistically significant (p = 0.003). Platelet levels on the 10th day of the chemotherapy cycle were lower than pre GM-CSF levels, however were not statistically significant (p = 0.097). Post GM-CSF TNF-alpha and IL-6 levels were higher than pre GM-CSF levels. This difference was statistically significant for TNF-alpha (p = 0.002) however for IL-6 a statistically significant difference was not detected (p = 0.55). GM-CSF does not significantly effect IL-6 levels in contrast to TNF-alpha. CONCLUSION: Clinical implications of increased levels of TNF-alpha are unclear and for a precise determination further studies are needed.     

A repertoire of cytokines in human seminal plasma

The pathophysiological significance of seminal cytokines in sperm function is still controversial. We determined the repertoire of cytokines in seminal plasma obtained from men with or without abnormalities in semen and assessed the pathophysiological significance of seminal cytokines. After conventional analysis of semen samples obtained from 86 men, levels of seminal cytokines (interleukin [IL]-1alpha, IL-2, IL-4, IL-6, IL-8, tumor necrosis factor-alpha [TNF-alpha], interferon-gamma, granulocyte colony-stimulating factor [G-CSF], macrophage CFS [M-CSF]) and granulocyte elastase were measured by an enzyme-linked immunosorbent assay. Leukocytospermia was defined as seminal plasma, which has > or =1000 ng/ml granulocyte elastase. Leukocytospermia was found in nine of 62 of the subjects in the normozoospermic group but in none of the 24 subjects showing abnormal sperm parameters (azoospermia, n=5; oligozoospermia, n=4; asthenozoospermia, n=15). The IL-8 level in the leukocytospermic group was significantly higher than those in the normal and oligozoospermic groups. IL-1alpha and TNF-alpha levels in the leukocytospermic group were significantly higher than those in the normal and asthenozoospermic groups. Although the G-CSF level in the leukocytospermic group was significantly higher than that in the normal group, high levels of M-CSF were detected in all groups. The IL-8 level was strongly correlated with IL-1alpha (r=0.935, P<0.0001) and G-CSF (r=0.916, P<0.0001) levels. Cytokines detected in seminal plasma are associated with the pathogenesis of leukocytospermia but not with the pathogenesis of asthenozoospermia and oligozoospermia.     

Amniotic fluid and maternal blood characteristics in severe mid-trimester twin-twin transfusion syndrome

OBJECTIVE: To investigate the concentrations of metabolic variables in the amniotic fluid of the recipient twin and maternal blood and to correlate them with clinical features, which are characteristic for the course of pregnancies with twin-twin transfusion syndrome (TTS). MATERIALS AND METHODS: In 109 pregnancies with severe mid-trimester TTS, we measured the concentrations of interleukin-6 (IL-6), alpha-fetoprotein (AFP), sodium, potassium, total protein, beta2-microglobulin and osmolality in the amniotic fluid of the recipient twin prior to laser coagulation of the vascular anastomoses and compared them to a control group of 92 singleton pregnancies. We measured the pulsatility index (PI) of ductus venosus flow velocity waveforms in the recipient twin and performed a retrospective chart analysis for complete maternal blood count before and after the procedure. RESULTS: All variables, except osmolality, IL-6 and AFP were significantly lower in the TTS group. There was a significant positive correlation between PI in the ductus venosus and the amniotic fluid AFP concentrations (r = 0.355; p < 0.001). There was a significant fall in complete maternal blood count after laser therapy (p < 0.001) and a significant correlation between the amount of amniotic fluid drained and the fall of maternal Hb (r = -0.261; p = 0.03) and hematocrit (r = -0.212; p = 0.01). CONCLUSION: Pathophysiologic changes in TTS do not only occur at the inter-twin level of placental vascular anastomoses but also at the materno-fetal level of fluid exchanges. AFP is correlated to the severity of congestive heart failure of the recipient twin.     

May GM-CSF influence the ability to fertilize oocytes?

The aim of this work was to evaluate the concentration of granulocyte-macrophage colony stimulating factor (GM-CSF), IL-1 beta IL-6 and TNF alpha in follicular fluid obtained from mature oocytes during in vitro fertilization scheme and to determine a correlation between GM-CSF concentration and the analyzed cytokines. Material consisted of 36 samples of follicular fluid which were divided into two groups: the first one--17 samples of follicular fluid obtained from follicles with successful oocyte fertilization and cleavage and the second one--19 samples of follicular fluid with unsuccessful fertilization. The concentration of GM-CSF as well as IL-6, IL-1 beta and TNF alpha in follicular fluid was evaluated by using the ELISA immunosorbent assays. The mean concentration of GM-CSF, IL-6, IL-1 beta was similar in both groups, however, the frequency of occurrence of these parameters as well as coexistence of GM-CSF with analyzed cytokines were higher in the second group i.e. in the follicular fluid with unsuccessful fertilization. We found a positive correlation between concentration of GM-CSF and IL-6 in follicular fluid of all samples. The authors suggested that activation of macrophages to an increased release of GM-CSF during disturbance of oocyte's maturation might implicate a relation between immune and endocrine systems with cytokine mediation.     

Increased plasma soluble fms-like tyrosine kinase 1 and endoglin levels in pregnancies complicated with preeclampsia

Background.?Increased maternal plasma levels of proinflammatory cytokines as well as the anti-angiogenic agents soluble fms-like tyrosine kinase 1 (sFlt-1) and endoglin (sEng) are associated with promoting vascular dysfunction leading to the maternal syndrome of preeclampsia.

Objective and method.?Nulliparous women complicated with preeclampsia (n = 29) and their corresponding controls (n = 29) delivering at the Enrique C. Sotomayor Obstetrics and Gynecology Hospital, Guayaquil-Ecuador were requested to participate in a study evaluating plasma levels of soluble anti-angiogenic factors (sFlt-1 and sEng) and pro-inflammatory cytokines: interleukin 6 (IL-6), interleukin 8 (IL-8), granulocyte colony stimulating factor (G-CSF), and tumor necrosis factor-alpha (TNF-α). Maternal and neonatal data were also assessed and compared among the study groups.

Results.?No significant differences in either maternal baseline or delivery characteristics were observed among the study groups. Compared with controls, preeclamptic women exhibited higher plasma levels of sFlt-1 (19.0 ± 15.1 vs. 12 ± 8.3 ng/mL) and of sEng (20.4 ± 9.9 vs.15.9 ± 9.4 ng/mL); respectively, p < 0.05. Women with severe disease displayed higher sFlt-1 and sEng levels when compared with mild ones (34.5 ± 11.6 vs. 9.5 ± 1.6 ng/mL, and 29.5 ± 9.0 vs. 14.8. ± 5.2 ng/mL, respectively; p < 0.001). In contrast, women with preeclampsia exhibited significant lower IL-8 and G-CSF levels compared with controls. No differences existed between either group in IL-6 levels or TNF-α.

Conclusion.?Consistent with previous reports, increased sFlt-1 and Eng levels in maternal plasma is consistent with vascular dysfunction found in gestations complicated with preeclampsia.     

Peritoneal fluid from endometriosis patients switches differentiation of monocytes from dendritic cells to macrophages

Immunological abnormalities of cell-mediated and humoral immunity might be associated with the pathogenesis of endometriosis. This study has examined the effects of peritoneal fluid obtained from patients with endometriosis (ePF) on the phenotypic characteristics of macrophages and dendritic cells (DCs) derived from monocytes. Monocytes were obtained from healthy young volunteers and cultured with ePF (n=12) or a control PF (cPF) (n=5) in the presence or absence of macrophage-colony stimulating factor (M-CSF) or IL-4 plus granulocyte macrophage-colony stimulating factor (GM-CSF). The ePF was demonstrated to increase expression levels of CD14 and CD64 on isolated monocytes in the presence or absence of M-CSF. Compared with cPF, addition of 10% ePF to GM-CSF plus IL-4-treated monocytes significantly down-regulated CD1a expression and up-regulated CD64 expression, but did not enhance expression levels of class II MHC. ePF had no effect, however, on tumor necrosis factor-alpha-induced maturation of DC. Levels of IL-6, IL-10 and M-CSF production were higher in ePF-treated than cPF-treated monocytes for both cell culture conditions with GM-CSF plus IL-4 and M-CSF. A neutralizing IL-6 antibody, but not an IL-10 antibody, abrogated the ePF-induced down-regulation of CD1a, up-regulation of CD64 and secretion of M-CSF. These results suggest that ePF favorably induces monocyte differentiation toward macrophages rather than DCs, and that this effect is mediated by IL-6. A reciprocal mode of cell differentiation between macrophages and DCs in response to ePF may be related to the pathogenesis of endometriosis.     

Secretion of cytokines by villous cytotrophoblast and extravillous trophoblast in the first trimester of human pregnancy

Cytokines are proposed to play roles in regulation of trophoblast invasion, spiral artery remodeling and immunoregulation during early pregnancy. Secretion of 12 cytokines (interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p70, IL-13, IFNγ, GM-CSF, MCP-1 and RANTES) by first trimester extravillous trophoblast and villous cytotrophoblast cells was examined using multiplex cytokine array technology. Seven (IL-1β, IL-8, IL-12p70, IL-13, GM-CSF, MCP-1 and RANTES) of the 12 cytokines examined were detectable in the samples studied (n=10 each group). Villous cytotrophoblast production of IL-1β and IL-8 increased with gestational age. Extravillous trophoblast production of IL-8, IL-13 and RANTES increased with gestational age. At 12-14 weeks gestation extravillous trophoblast cells secreted higher levels of IL-8, IL-13 and RANTES than villous cytotrophoblast cells.