Higher levels of soluble E-cadherin in cyst fluid from malignant ovarian tumours than in benign cysts.

A major diagnostic dilemma in the clinical gynaecological oncology setting is to preoperatively determine whether a complex ovarian mass is benign or malignant. The cell-cell adhesion molecule E-cadherin has previously been localised in biopsies from both benign and malignant epithelial ovarian tumours. In this study, soluble E-cadherin levels was measured with ELISA-technique in peripheral blood, ascites and cystic fluids from patients (n = 33) undergoing surgery for ovarian cystic masses. The levels of soluble E-cadherin were significantly higher in cystic fluid from cystadenocarcinomas (p < 0.001) and borderline tumours (p < 0.05) as compared to cystic fluid from cystadenomas. In ascites fluid and peripheral blood no significant differences were seen. However, ratios of cystic fluid/peripheral blood levels were significantly higher in cystadenocarcinoma (p < 0.001) and borderline tumours (p < 0.05) as compared to benign tumours. In conclusion, measurements of soluble E-cadherin in cystic fluid from patients presenting with complex ovarian masses may be beneficial in increasing the accuracy of preoperative diagnosis.

The chemotactic cytokine interleukin-8--a cyst fluid marker for malignant epithelial ovarian cancer?

Due to the difficulties in separating malignant and benign ovarian cysts by transvaginal ultrasound and other techniques, there is a need for biochemical markers in serum or cyst fluids. In the present study we have evaluated the levels of the chemokine interleukin-8 (IL-8) in ovarian cysts. IL-8 is known to be expressed in the normal ovary and to influence proliferation and angiogenesis of several nonovarian types of tumors. Cyst fluids from benign (n = 15) and malignant (n = 13) ovarian tumors were analyzed. The levels of IL-8 were found to be significantly (13-fold) higher in cyst fluids from malignant tumors (18.1 +/- 7.5 ng/ml; mean +/- SE) compared to benign cysts (1.3 +/- 0.7 ng/ml). The plasma levels of IL-8 were considerably lower (2.9 and 0.3% of levels in benign and malignant cyst fluids, respectively) than in cyst fluids. No difference in the plasma levels of patients with benign or malignant tumor could be detected. In contrast, the levels of CA 125 were significantly higher in plasma of patients with malignant disease with the inverse relation in cyst fluids. In conclusion, the levels of IL-8 are markedly elevated in cyst fluid from malignant tumors compared to benign. This specific increase indicates a role for this cytokine in ovarian tumor biology.

Increased release of tumor necrosis factor-alpha and interleukin-6 in women with the syndrome of hemolysis, elevated liver enzymes, and low platelet count.

BACKGROUND: Complement is activated in preeclampsia and complement products are known to activate macrophages. The aim of this study was to determine whether the macrophage derived cytokines, interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha, are released in patients with a form of severe preeclampsia characterized by the syndrome of hemolysis, elevated liver enzymes, and low platelet count (HELLP syndrome). METHODS: Complement activation and plasma levels of cytokines were studied in 11 women with HELLP syndrome and in 11 controls with uncomplicated pregnancies. To further evaluate the connection between complement activation and cytokine release an in vitro study on heparinized whole blood incubated with recombinant C5a was performed. RESULTS: In the HELLP group, complement anaphylatoxin C5a was increased in plasma at delivery (p < 0.01) and one day after delivery (p < 0.05), terminal C5b-9 complement complex was elevated in plasma at delivery (p < 0.001) and one day after (p < 0.01), plasma levels of interleukin-6 were increased one day after delivery (p < 0.01), and plasma concentrations of tumor necrosis factor-alpha were elevated at delivery (p < 0.01), compared with corresponding levels in controls. All parameters normalized within one week. Interleukin-I beta did not differ between the groups. In vitro, recombinant C5a incubated in whole blood gave a dose-dependent release of interleukin-6. No increased release of interleukin-1 or tumor necrosis factor-alpha was seen after incubation. CONCLUSIONS: Since cytokine release occurs in severe preeclampsia, inflammatory mechanisms may participate in the pathophysiology of severe preeclampsia.

Increased concentrations of neopterin in plasma, ascites and ovarian cyst fluid in malignant tumours compared with benign ovarian tumours.

Neopterin is a marker of the activation of cell-mediated immunity. The aim was to determine whether the concentrations of neopterin differ in plasma, ascites and ovarian cyst fluid between patients with ovarian cancer and patients with benign ovarian tumours. Neopterin was measured in 29 patients with cystic ovarian tumours of unknown histology. 14 ovarian cancers and 15 benign ovarian tumours were diagnosed histologically. Patient age and tumour size did not differ significantly between the groups. Neopterin levels were determined in plasma, and in ascites and cyst fluid by ELISA. The neopterin concentration in plasma, ascites and ovarian cyst fluid was significantly higher in patients with ovarian cancer compared with patients with benign ovarian tumours of the same size. The study shows that activation of cell-mediated immunity, defined as increased formation of neopterin, was increased in patients with ovarian cancer compared with patients with benign ovarian tumours.

Tissue Trauma after Laparoscopic and Abdominal Hysterectomy Measuring Inflammatory Response

It is suggested that laparoscopic surgery reduces postoperative pain and shortens hospital stay and convalescence because of the small amount of tissue trauma. We evaluated the inflammatory response during abdominal hysterectomy (AH, 12 women) and laparoscopic hysterectomy (LH, 12 women) by measuring interleukin (IL)-6, neopterin and terminal C5b9 complement complex (TCC). Blood samples were drawn preoperatively, perioperatively, 1 minute, 24 hours, and 7 days postoperatively. Levels of IL-6 were determined to evaluate cytokine release, neopterin was determined as a marker for macrophage-monocyte activation, and TCC was determined to assess complement activation. The IL-6 concentrations, as a percentage of preoperative level, were significantly elevated postoperatively in both groups, and also perioperatively in the LH group. Neopterin concentrations, as a percentage of perioperative level, were significantly increased in the LH group preoperatively and postoperatively. No elevation was seen in the AH group. There was no sign of complement activation in either group. Our results indicate significant tissue trauma during both LH and AH. The extent of trauma might be greater in laparoscopic surgery. Despite this, the LH group had a shorter hospital stay and convalescence than the AH group. The proposed advantages to the patient of laparsocopic surgery thus seem to be attributable to other factors than the amount of tissue trauma.

Evaluation of tissue trauma after laparoscopic and abdominal hysterectomy: measurements of neutrophil activation and release of interleukin-6, cortisol, and C-reactive protein.

BACKGROUND: Trauma and major surgery stimulate a cascade of events that mediate the inflammatory response. The aim of our study was to determine whether or not hysterectomy leads to release of cytokines, cortisol, and C-reactive protein (CRP), activation of neutrophils, and activation of the complement cascade. A further aim was to compare laparoscopic and abdominal hysterectomy with regard to the same parameters. STUDY DESIGN: Twenty-four consecutive patients were randomized to either abdominal (n = 12) or laparoscopic hysterectomy (n = 12). Blood samples were drawn preoperatively, intraoperatively, and then at one minute, 24 hours, and seven days postoperatively. Interleukin-6 (IL-6) levels were used to evaluate cytokine release, cortisol and CRP to evaluate the inflammatory response, and polymorphonuclear (PMN) elastase to detect neutrophil activation. To evaluate complement activation, the terminal C5b-9 complement complex (TCC) was determined. RESULTS: Interleukin-6 concentrations were significantly elevated one minute and 24 hours postoperatively in both groups. Independent of the surgical technique or operative time, the highest IL-6 concentration was reached four hours after beginning the operation. Cortisol levels were significantly elevated during and after the operation in both groups. C-reactive peptide levels were significantly elevated in both groups 24 hours and seven days after the operation. Polymorphonuclear elastase was elevated 24 hours postoperatively in both groups. There were no signs of complement activation during the operative period or postoperatively in either patient group. CONCLUSIONS: Our results indicate serious tissue trauma during both laparoscopic and abdominal hysterectomy. The extent of surgical trauma did not differ between the two operative methods.

Release of inflammatory mediators in association with collection of wound drainage blood during orthopaedic surgery.

Ten patients undergoing hip replacement surgery were studied regarding activation of complement and leukocytes in association with collection of wound drainage blood. The blood was collected postoperatively but not reinfused due to the possible risks with reinfusion of blood containing inflammatory mediators. Blood samples for analysis of complement activation (TCC), leukocyte activation (PMN elastase) and cytokines (Interleukin-6) were drawn preoperatively from the patients. Blood samples were also drawn intraoperatively from the wound. Samples were also drawn from the collected wound drainage blood, before and after blood was passed through a microporous filter. There were elevated concentrations of TCC, PMN elastase and IL-6 in the collected wound drainage blood before and after the filter. The filtration did not significantly reduce the concentrations of these factors. In the wound blood the concentrations were higher compared to those found in the systemic blood preoperatively, but lower compared to concentrations found in the collected drainage blood. The study demonstrates that the collection of wound drainage whole blood is associated with activation of complement, release of PMN elastase and cytokines.

The role of complement in pregnancy-induced hypertensive disease.

Preeclampsia is a pregnancy-induced hypertensive disease with an incidence of about 5% in primigravidas and, being common, it significantly contributes to maternal and neonatal morbidity and mortality. The primary cause remains unknown but might be immunologic, since immunologic aberrations are described in preeclampsia. Activation of the complement system in pregnancy-induced hypertensive disease has been discussed during the last 60 years. It is now strongly indicated that complement activation occurs in preeclampsia. The complement system is very potent and one of the major effector pathways of the process of inflammation. The pathological manifestations, endothelial damage and microvascular injury, and thereby the clinical findings in preeclampsia, may be explained by complement activation, resulting in the direct vascular effects of biological active complement components and complement-mediated activation of leukocytes, with release of potent inflammatory mediators. This new etiological hypothesis might give other options in therapy and prevention of pregnancy-induced hypertensive disease.

Complement, neutrophil, and macrophage activation in women with severe preeclampsia and the syndrome of hemolysis, elevated liver enzymes, and low platelet count.

Activation of complement, neutrophils, and macrophages was studied in 14 women with severe preeclampsia, 11 of whom had the syndrome of hemolysis, elevated liver enzymes, and low platelet count; in 14 women with normal pregnancies; in seven normal pregnant women undergoing cesarean deliveries; and in 15 healthy nonpregnant women. Activation of complement, neutrophils, and macrophages was measured by plasma determinations of complement split products, polymorphonuclear (PMN) elastase, and neopterin, respectively. Women with severe preeclampsia had increased levels of C5a, terminal complement complex, PMN elastase, and neopterin at delivery and 1 day postpartum as compared with the normal pregnant group. One week postpartum, neopterin remained higher in preeclamptic women, whereas the complement components and PMN elastase had returned to normal. Cesarean delivery after normal pregnancy did not increase the levels of complement split products, PMN elastase (except for one value), or neopterin. The nonpregnant women had normal PMN elastase and neopterin levels. Accordingly, complement, neutrophils, and macrophages are activated in women with severe preeclampsia at delivery. The plasma levels of PMN elastase correlated positively to the formed terminal complement complexes in vivo. An in vitro study was performed to elucidate further the connection between complement and leukocyte activation. Recombinant C5a incubated in whole blood and in a neutrophil cell suspension gave a dose-dependent release of PMN elastase. Both the clinical and the in vitro results indicate that activation of the complement system may affect the function of neutrophils. This study supports the theory that the pathologic manifestations of severe preeclampsia may be explained by complement-induced release of biologically active substances from activated leukocytes.

Opinions on treatment of women with habitual abortion based on investigations for blocking antibody and autoantibodies.

Three hundred and thirty-seven women with habitual abortion of unknown etiology were studied for cellular reactivity and blocking antibody in one-way mixed lymphocyte culture. Their sera were investigated for anti-cardiolipin antibodies, antinuclear antibodies, and antibodies against DNA, and the activated partial thromboplastin time (APTT) and complement levels of their plasma were determined. Increased anti-cardiolipin antibody levels were demonstrated in 77 (22%) of the 337 women, all of whom were considered healthy and had no signs of autoimmune disease. Most patients with high anti-cardiolipin antibody levels displayed lowered values of complement factor C4. According to our experiences, the mere occurrence of anti-cardiolipin antibody in women with habitual abortion is no absolute cause for treatment with prednisolone, not even in cases with greatly elevated anti-cardiolipin values. Therapy with prednisolone and acethylsalicylic acid (ASA) during pregnancy should be given to those women who have high levels of anti-cardiolipin antibodies concomitant with high APTT values, low values of complement C4, and strong blocking antibody. Anti-cardiolipin antibody has been investigated during pregnancy in 136 normal pregnant women, 11 of whom (8%) were positive at any sampling occasion, but only one of whom (1%) had high levels. Evidently the development of anti-cardiolipin antibody is no normal feature of pregnancy among Swedish women and thus the high frequency found among healthy Swedish women with habitual abortion remains unexplained. We have introduced an immunization program of leukocyte transfusions in habitual abortion. The development of previously absent blocking antibody seems to be a valuable prognostic sign of possible success for immunization therapy against habitual abortion.

Complement activation in relation to development of preeclampsia.

Six hundred eighty-five primigravidas followed as a series had complement activation evaluated by the formation of anaphylatoxins (C3a and C5a) and terminal C5b-9 complement complexes in venous blood. Samples for complement determinations were obtained four times during pregnancy, in pregnancy weeks 12-16, 20-24, 28-32, and 34-36. Seven of the women developed preeclampsia and one of them the syndrome of hemolysis, elevated liver enzymes, and low platelet count (HELLP syndrome). Eleven others with uncomplicated pregnancies were selected as a control group. Plasma samples were taken from these 18 women at delivery and 1 and 7 days after delivery. At delivery, plasma C5a levels were significantly greater in the preeclamptics than in controls, and four of the seven preeclamptics had elevated plasma C3a values compared with controls. One week after delivery, these plasma anaphylatoxins had returned to normal. Elevations of the anaphylatoxins could not be detected before the women developed clinical signs of preeclampsia. No alterations in terminal C5b-9 complement complexes could be observed in the women with preeclampsia. However, the women who developed HELLP syndrome had elevated plasma concentrations of C3a, C5a, and terminal C5b-9 complement complex at delivery. These values returned to the normal range 1 week after delivery. We conclude that complement activation in the systemic circulation does not occur early in pregnancy and that plasma concentrations of C3a, C5a, or terminal C5b-9 complement complex cannot be used as predictors of preeclampsia.

Enhanced anaphylatoxin and terminal C5b-9 complement complex formation in patients with the syndrome of hemolysis, elevated liver enzymes, and low platelet count.

Complement activation was studied in ten consecutive pregnant women developing hemolysis, elevated liver enzymes, and low platelet count (HELLP syndrome) and ten other women with normal pregnancies. Blood samples for anaphylatoxin (C3a/C3a desArg and C5a/C5a desArg) and terminal C5b-9 complement complex determinations were drawn at delivery and 24 hours and 7 days later. Women developing HELLP syndrome had higher plasma levels of anaphylatoxins with delivery than did women with uneventful pregnancies. The plasma levels of terminal C5b-9 complement complexes at the time of delivery were increased as compared with levels 1 and 7 days after delivery in women with HELLP syndrome. The plasma concentrations of the anaphylatoxins and the terminal C5b-9 complement complexes returned to normal levels within 1 week after delivery in the HELLP group. The formation of C5b-9 complement complex indicates that the terminal part of the complement cascade has been activated and that C5a has been formed and eliminated. Complement activation with release of anaphylatoxins and terminal C5b-9 complement complexes may be one etiologic factor behind the elevated blood pressure, hemolysis, liver insufficiency, and platelet consumption seen in these patients.

The results of investigations for blocking antibody and autoantibodies help to choose between different immunological treatments for recurrent abortion.

Immunologic investigations were performed on 337 healthy women with unexplained habitual abortion. Their sera were investigated for antinuclear antibodies (ANA), antibodies against DNA, and anti-cardiolipin antibodies, and the activated partial thromboplastin time (APTT) and complement levels of their plasma were determined. Cellular reactivity and blocking antibody were studied in one-way mixed lymphocyte culture (MLC). None of the women had any signs of autoimmune disease. However, in 77 of the 337 women (22%) increased anti-cardiolipin antibody levels were found, in 19 (7%) above 10 units. Most patients with high anti-cardiolipin antibody levels had lowered values of complement factor C4. We consider the mere occurrence of anti-cardiolipin antibody in women with habitual abortion to be no absolute cause for treatment with prednisolone, and this is true even in cases with very elevated anti-cardiolipin values. Treatment with prednisolone and acetylsalicylic acid (ASA) during pregnancy should be given to only those women who have high levels of anti-cardiolipin antibodies concomitant with high APTT values, and low values of complement C4. Anti-cardiolipin antibody was also investigated during pregnancy in 136 normal pregnant women. Eleven of them (8%) were positive at any of four sampling occasions, but only one (1%) had high levels. These data differ significantly from those of the habitual aborters. Thus the development of anti-cardiolipin antibody is no normal feature of pregnancy in Swedish women and so the high frequency found among healthy Swedish women with habitual abortion remains unexplained. We have introduced an immunization programme of third party leukocyte transfusions in habitual abortion.(ABSTRACT TRUNCATED AT 250 WORDS)

Complement activation and anaphylatoxin (C3a and C5a) formation in preeclampsia and by amniotic fluid.

The aim of this study was to determine whether the biologically active complement peptides C3a and C5a are formed in pregnancy and whether amniotic fluid can activate complement. C3a and C5a are formed when complement is activated. They increase smooth-muscle contraction, vascular permeability, and histamine release from mast cells and basophils. Thirty pregnant women were studied, 16 with uncomplicated and 14 with preeclamptic pregnancies. The plasma C3a and C5a concentrations before delivery were significantly higher in the preeclamptic than in the normal group. The concentrations returned to normal within 1 week. Plasma, serum, and amniotic fluid from 12 pregnant women (eight uncomplicated and four preeclamptic pregnancies) were drawn in connection with delivery. Amniotic fluid was incubated in fresh autologous serum at 37C for 15 minutes. A dose-dependent formation of C3a and C5a was registered with increasing amounts of amniotic fluid.