Production of cytokines during interaction of peripheral blood mononuclear cells with autologous ovarian cancer cells or benign ovarian tumour cells.

Cytokines produced by tumour and immune cells may play a significant role in a modulation of immune cells response against tumour. We investigated an ability of peripheral blood mononuclear cells (PBMC) of patients with early and advanced stages of ovarian cancer and from non-cancer patients to produce various cytokines in the presence or absence of autologous ovarian cancer (OC) cells or benign ovarian tumour (BOT) cells. Activated PBMC of patients with advanced stage of cancer produced slight amount of interferon gamma (IFN-gamma) and what's more, the production of IFN-gamma was decreased in the presence of OC cells. PBMC of patients with ovarian cancer or benign ovarian tumour generated comparable amounts of interleukin 6 and 10 (IL-6, IL-10), and transforming growth factor beta1 (TGF-beta1). PBMC of the patients with cancer produced higher amount of tumour necrosis factor alpha (TNF-alpha) than PBMC of non-cancer patients. We demonstrated here that the reciprocal contact of OC cells from advanced cancer with autologous PBMC altered the direction of produced cytokines and leads to the down-regulation of IFN-gamma and TNF-alpha as well as to up-regulation of immunosuppressive (IL-10, TGF-beta1) and pro-inflammatory (IL-6) cytokines production.

Frequencies of killer immunoglobulin-like receptor genotypes influence susceptibility to spontaneous abortion.

Natural killer (NK) cells are the most abundant lymphocyte population in the decidua. These cells express killer immunoglobulin-like receptors (KIRs), which upon recognition of HLA class I molecules on trophoblasts may either stimulate NK cells (activating KIRs) or inhibit them (inhibitory KIRs) to produce soluble factors necessary for the maintenance of pregnancy. KIR genes exhibit extensive haplotype polymorphism; individuals differ in both the number and kind (activating vs. inhibitory) of KIR genes. This polymorphism affects NK cell reactivity and susceptibility to diseases, including gynecological disorders. Therefore we KIR-genotyped 149 spontaneously aborting women and 117 control multiparae (at least 2 healthy-born children). Several genotypes (i.e. combinations of various KIR genes) were differently distributed among the patients and control subjects. Differences were observed in the numbers and the ratios of activating to inhibitory KIRs between patients and healthy women: (i) genotypes containing 6 activating KIR genes were less frequent and those containing 6 inhibitory KIR genes were more frequent in patients than in control subjects, and (ii) an excess of inhibitory KIRs (activating-to-inhibitory KIR gene ratios of 0.33 to 0.83) was associated with miscarriage, whereas ratios close to equilibrium (0.86-1.25) seemed to be protective. In addition, the results suggest for the first time that sporadic and recurrent spontaneous abortions as well as miscarriage in the presence or absence of autoantibodies may have different KIR genotypic backgrounds.

Involvement of innate immunity in the pathogenesis of chronic hepatitis C in children.

The aim of the study was to assess the role of selected elements of innate immunity in the pathogenesis of chronic hepatitis C in children. The study comprised 20 children with chronic hepatitis C (group 1), nine healthy hepatitis C virus (HCV) seropositive children (group 2) and 18 healthy children (control group). We evaluated the expression of Toll-like receptor (TLR)2 and TLR4 on peripheral blood neutrophils, and generation of interleukin (IL)-8, IL-10, IL-12 and reactive oxygen species (ROS) by neutrophils. The performed tests demonstrated higher expression of TLR2 and TLR4 on stimulated neutrophils and of TLR4 on non-stimulated neutrophils in group 1 in comparison to HCV seropositive children and controls. In group 1, the expression of TLR2 after granulocyte colony-stimulating factor (GCSF) stimulation showed positive correlation with alanine aminotransferase and asparate aminotransferase activities, while the expression of TLR2 without stimulation and of TLR4 after GCSF stimulation also correlated with necrosis. IL-12 generation by lipopolysachcharide-stimulated neutrophils was higher in group 1 versus controls. In group 1, maximum chemiluminescence (CL) without pre-activation, both spontaneous and after formyl-methionyl-leucyl-phenylanine and phorbol-myristate-acetate (PMA) stimulation, was significantly lower than in the controls. CL after tumour necrosis factor-alpha pre-activation and PMA stimulation was still lower than in the controls, however, after opsonized zymosane stimulation it was significantly higher than in the controls. Our studies suggest the involvement of neutrophils in the pathogenesis of chronic hepatitis C in children. Neutrophils demonstrate increased expression of TLR2 and TLR4 (correlating with the features of hepatocytic damage and intensification of necrosis), inhibition of oxygen metabolism, and after TNF-alpha pre-activation higher ability to produce ROS.

'In vitro' cytokine secretion by peripheral blood and decidual lymphocytes during the third trimester of normal pregnancy.

If the 'Th2 phenomenon' is dependent on trophoblastic antigens and cytokines, the profile of cytokines secreted by decidual lymphocytes (DL) should indicate stronger Th2 shift than that of peripheral blood lymphocytes (PBL). We studied spontaneous and mitogen-stimulated 'in vitro' cytokine secretion of cultured lymphocytes isolated from peripheral blood (n = 21) and decidua (n = 11) of third trimester healthy pregnant women not being in labor. The ELISA method was used for estimation of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma and TGF-beta. The results (given in pg/ml) were found to be statistically significant in the Mann-Whitney U-test (p < 0.05). Compared to PBL, DL produced spontaneously decreased concentrations of IL-12, IL-6, IFN-gamma and TGF-beta and upon mitogen stimulation increased quantities of IL-2, IL-4 and IL-10. It demonstrates that DL play an active role in local Th2-like immunoregulation and that their cytokine secretion pattern estimated in 'in vitro' conditions is biased towards Th2 activity more than that of PBL.

The effect of LPS on neutrophils from patients with high risk of type 1 diabetes mellitus in relation to IL-8, IL-10 and IL-12 production and apoptosis in vitro.

Innate immunity includes neutrophil inflammatory function, tissue destruction and regulatory cytokine production. Programmed cell death (apoptosis) is postulated to be a key mechanism for neutrophil elimination during inflammation. The aim of the present study was to evaluate the neutrophil apoptosis in relation to IL-8, IL-10 and IL-12 production in vitro by neutrophils of patients suffering from diabetes mellitus (DM)1 and the first-degree relatives of patients with DM1. The early stage of neutrophils apoptosis was assessed morphologically, and the later stage by DNA-binding dye propidium iodide, both after treatment with lipopolysaccharide (LPS), insulin or anti-CD95 antibody (Ab) as stimulators. CD16 (FcgammaRIII) receptor expression was also evaluated. Production of IL-8, IL-10, and IL-12 cytokine was evaluated in supernatant after neutrophil incubation for 21 h in culture medium alone, in medium in the presence of LPS, insulin or anti-CD95 antibody (Ab). Cytokine concentrations were measured by enzyme-linked immunosorbent assay (ELISA) method using commercially available kits. Our study demonstrates that LPS inhibits the early stage of apoptosis (as evaluated morphologically) of healthy donors' neutrophils. The LPS-dependent early apoptosis inhibition of neutrophil of patients with DM1 or in prediabetics was decreased in comparison with control. The later stage of apoptosis of neutrophils treated in vitro with anti-CD95 Ab of patients suffering from DM1 was decreased in comparison with prediabetics and healthy donors (propidium iodide (PI) staining). LPS-induced production of anti-apoptotic cytokines IL-8, IL-10 by neutrophils of prediabetic and patients with DM1 was increased. The formyl-methionyl-leucyl-phenylalanine (fMLP)-induced proapoptotic reactive oxygen intermediates (ROI) production was significantly higher in DM1 patients. We have concluded that neutrophils from prediabetic and diabetic patients demonstrated the misbalance in anti-apoptotic IL-8 and IL-10 cytokine and proapoptotic ROI production. LPS-dependent IL-12 overproduction by neutrophils is responsible for the switch in T helper Th1/Th2 balance to Th1 and in this way may participate in inflammation and autoimmune DM1 progression.

[The use of flow cytometry for the determination of the hepatitis B virus pre-S1 envelop protein binding by human peripheral blood lymphocytes in vitro in the course of recovery]. / Zastosowanie cytometrii przeplywowej do oceny wiazania fragmentu Pre-S1 antygenu powierzchniowego wirusa zapalenia watroby typu B przez limfocyty ludzkiej krwi obwodowej in vitro u osób po przebyciu infekcji.

The course of hepatitis B virus (HBV) infection depends on the host immune response to various antigens of the virus, including hepatitis B surface antigen (HbsAg), which is present on the envelope of HBV and which contains Pre-S1, Pre-S2 and S proteins. The peripheral blood lymphocytes (PBL) from patients with chronic hepatitis B infection (CHB) and patients acutely infected with HBV and recovered completely (convalescents) were studied for detecting of Pre-S1 antigen binding. We used a cytometric method for measurement of the binding of fluorescein labeled Pre-S1 antigen (FITC/Pre-S1) to the PBL. The binding of FITC/Pre-S1 was determined on resting lymphocytes and on lymphocytes cultured in vitro for 5 days in the presence of Pre-S1 and phytohaemagglutinin-P (PHA-P). The expression of CD3 and CD19 molecules on the surface of PBLs simultaneously with the expression of FITC/Pre-S1 was also analysed using flow cytometry. We found that the Pre-S1 binding significantly depends on the state of the activation of lymphocytes. Specific stimulation of the convalescent lymphocytes with Pre-S1 resulted in an increase in the percentage of the FITC/Pre-S1 binding cells in relation to the unstimulated cultured and resting cells as well as to the cells from CHB patients. We suggest that flow cytometric measurement of Pre-S1 binding by lymphocytes may be useful indicator of the disease activity.

[Current opinions on immunological processes in rheumatoid arthritis during pregnancy]. / Wspólczesne poglady na przebieg procesów immunologicznych w reumatoidalnym zapaleniu stawów u kobiet w ciazy.

The essential in pathogenesis of RA is induction of incorrect immunological response against synovial and connective tissue antigens, which depends of CD4+ T-cells activation by specific antigen. This stimulation leads to releasing Th1 lymphokines. The most important cytokine is TNF-alpha. An increased level of TNF-alpha, IL-1, IL-6, GM-CSF, IL-8 was observed in patients with RA. PDGF, FGF, TGF, C-X-C a chemokines (IL-GRO-alpha, ENA78) and CCb chemokines (RANTES, MCP1 MIP1 alpha) are also involved in synovial hyperplasia in RA. During a pregnancy a clinical improvement in women with RA is frequent. The reason of this fact is probably connected with Th2 predominance (IL-4, IL-10) caused by presence of fetal tissues. Specific, cell-mediated immunity is suppressed and changed to Th2 by progesterone and PGE2. During a pregnancy a higher sensitivity of lymphocytes to progesterone was found. Progesterone stimulates T cells to PIBF production, which decreases NK activity. Th2 cytokines (Il-6, IL-10, IL-13, TGF) are expressed on decidua and inhibit secretion of Th1 cytokines (IL-2, INF gamma, TNF-alpha, IL-1 alpha, IL-1 beta). Immunosuppression caused by pregnancy probably decreases inflammatory and destructive reactions in tissues women with RA. The first attack of this disease frequently observed during puerperium is connected with a high level of prolactin and a low of estrogens, which causes a increased release of IL-2 and has a main influence on initiation and increasing of inflammatory process in RA.

[Evaluation of peripheral blood neutrophils activity in women with endometriosis]. / Ocena aktywnosci neutrofilów krwi obwodowej u kobiet z endometrioza.

Authors evaluated the neutrophils ability of creating reactive oxygen intermediates (ROI) in peripheral blood of 14 patients with endometriosis. These tests were performed using the chemiluminescence method. Enhanced ROI production and in vivo priming phenomenon of neutrophils in endometriosis were reported. The observed increase in oxygen activity of neutrophils was statistically significant only in reaction with the use of fMLP. However performed neutrophils preactivation with TNF-alpha followed by the chemiluminescence test revealed the occurrence of in vivo neutrophil activation in women with endometriosis. Reported data confirm the existence of local inflammatory reaction in the course of endometriosis.

[Evaluation of the generation of interleukin-10 by peripheral blood lymphocytes in women with endometriosis]. / Ocena generacji interleukiny-10 przez limfocyty krwi obwodowej u kobiet z endometrioza.

The aim of study was the evaluation of generating the IL-10 by peripheral blood lymphocytes in women with endometriosis according to the stage of of the disease. Cytokine was estimated by immunoenzymatic method Elisa. The generation of IL-10 by peripheral blood lymphocytes was lower in patients with all stages of endometriosis than in control group. Decreased IL-10 generating observed in our study was not statistically significant, but it may correlate with disorders of immunological cell response. It may also play a role in a development and progression of endometriosis.

Activated T lymphocytes from patients with high risk of type I diabetes mellitus have different ability to produce interferon-gamma, interleukin-6 and interleukin-10 and undergo anti-CD95 induced apoptosis after insulin stimulation.

Type I Diabetes mellitus (DM1) is the effect of T cell dependent autoimmune destruction of insulin producing beta cells in the pancreas islet. T cells are activated in response to islet dominant autoantigens, the result being the development of DM1. Insulin is one of the islet autoantigens responsible for activation of T lymphocyte functions, inflammatory cytokine production and development of DM1. The experiments reported in this study have shown the spontaneous increase of CD95 molecule expression on lymphocytes of the first-degree relatives of DM1 patients. The autoantigen insulin is responsible for stimulation in vitro of potentially hazardous 'memory' lymphocytes to produce interleukin-6 (IL-6) and interleukin-10 (IL-10) interleukins. Insulin induced stimulation of lymphocytes in vitro was observed in patients at high risk of developing diabetes mellitus (prediabetics). Phytohaemagglutinin (PHA) stimulates lymphocytes of all groups in the same way. Stimulated lymphocytes in second cultures undergo apoptosis induced with anti-Fas specific antibodies. The deletion in vitro of resting peripheral lymphocytes is nonfunctional. Insulin activated T lymphocytes, which undergo apoptosis were not observed in peripheral blood of healthy people and in patients with DM1. This observation suggests that insulin is involved as autoantigen in DM1 progression in patients with high risk of diabetes type I. The autoreactive T lymphocytes may persist in peripheral blood of patients with high risk DM1. Defective elimination of autoreactive T cells may result in autodestructive damage of islets beta cells in the prediabetic stage and disease progression to DM1.

[Oxidative stress in pregnancies complicated by gestational hypertension (GH), effect of oxidized serum lipoproteins (O-LDL)--the role of antio-LDL antibodies]. / Nasilenie stresu oksydacyjnego w ciazach powiklanych nadcisnieniem ciazowym. Efekt dzialania utlenowanych lipoprotein osocza (O-LDL)--rola przeciwcial anty O-LDL.

OBJECTIVE: To explain correlations between metabolism of serous oxidized lipoproteins, their influence on placental function in pregnancies complicated by GH and intrauterine asphyxia. MATERIALS AND METHODS: The group of 233 pregnant women characterised by gestational age between 32-40 weeks were divided into two sub-groups: the study group of 105 women with GH and the control group of 128 healthy pregnant women. In all the subjects doppler flows in umbilical and middle brain artery as well as the serum levels of anti o-LDL antibodies (oLAB) were measured. RESULTS: In the study group mean PI values in middle brain artery equal 1.62 +/- 0.41 while in the control group 0.91 +/- 0.16 (p < 0.01), with oLAB values 1133 +/- 24 for the study group and 619 +/- 15 for the controls. Having analysed vascular flows in umbilical artery, mean PI values of 1.41 +/- 0.36 for the study group and 0.63 +/- 0.12 for the control group were obtained, (p < 0.01). The levels of oLAB in these groups were as follows: 1019 +/- 18 for the study group and 600 +/- 12 for the controls. CONCLUSIONS: 1. In pregnancies complicated by GH an intensified oxidative stress was observed as determined by increased levels of anti o-LDL antibodies (oLAB). 2. Pulsatility indices in umbilical and middle brain artery exhibited enhanced values in pregnancies complicated by GH.

[Parameters of peripheral whole blood neutrophil activity in women with pregnancy induced hypertension]. / Ocena niektórych parametrów aktywnosci ukladu neutrofili w pelnej krwi obwodowej ciezarnych z nadcisnieniem indukowanym ciaza.

OBJECTIVES: Pregnancy induced hypertension is believed to be a state of neutrophil overactivity, however all previous studies were done on isolated cells. DESIGN: To study neutrophil activity in whole blood of PIH women. MATERIALS AND METHODS: Neutrophil activity was estimated without isolation, in peripheral whole blood of 23 PIH women and 26 normal pregnant controls. The chemiluminescence test was performed without any stimulation and upon stimulation of neutrophils with fMLP, OZ and PMA, before and after pre-activation with TNF-alpha. The results were corrected according to the haemoglobin concentration and (%) of neutrophils. The percentage of whole blood neutrophils indicating expression of selectins CD18, CD11b, integrin CD62L and mean fluorescence intensity (MFI) of these molecules were studied on flow-cytometry. RESULTS: The study revealed that neutrophil chemiluminescence was not significantly higher in PIH women and after correction coefficient used it was even lower in PIH patients. TNF-alpha preactivation had no influence on chemiluminescence results. Expression of CD11b expressed as MFI value was significantly increased while that of CD62L, expressed as (%) of positive cells and MFI value--decreased in PIH patients. CONCLUSIONS: Changes of neutrophil CD11b and CD62 expression indicate increased activity of these cells in PIH women, however low production of reactive oxygen species estimated by corrected chemiluminescence test especially after TNF-alpha pre-activation, indicates that this form of their reactivity is rather "exhausted" during PIH.

[Decidual lymphocyte subsets in pregnant women]. / Subpopulacje limfocytów w doczesnej u ciezarnych z preeklampsja.

OBJECTIVES: Materno-foetal immunological reactions in decidua are probably one of the most important elements in pathogenesis of preeclampsia. DESIGN: To compare lymphocyte subsets isolated from decidua of preeclamptic pregnant women with lymphocyte subsets isolated from healthy pregnant women. MATERIALS AND METHODS: Preeclampsia (PE) was defined according to USA National Health Institute criteria. The study group consisted of 21 women with PE and 11 women with physiological pregnancy. All pregnancies were finished with elective cesarean sections. Exclusion criteria were: uterine contractions, infection, chorinamnionitis, diabetes mellitus and therapy with steroids less than 7 days before blood sampling. Decidual tissue was obtained by curettage of the uterine cavity. The fragments of decidua were separated from clotted blood and placed in sterile tubes with 5 ml of isotonic solution (PBS). Then the decidual tissue was mechanically fragmented, homogenized and rinsed in PBS. Routine immunofluorescent marking techniques with monoclonal antibodies were performed. Analysis was done with FACSCalibur flow-cytometer with 488 nm argon laser using CellQuest programme. The following lymphocyte subsets were estimated: CD3, CD19, CD4, CD8, CD4/CD29, CD8/CD28, CD4/CD45RA, CD4/CD45RO, CD56/CD16, CD69. The results were described as percentage of lymphocytes positive for above surface molecules. Statistical analysis was performed using t-Student and U-Mann-Whitney tests. The work was sponsored by KBN 4 P05E 118 15 grant. RESULTS: Decidua of pregnant PE women contains significantly increased percentage of CD3-/ CD56 + 16+, CD8+/CD28+ cells and decreased percentage of CD3+, CD19+, CD4+/CD29+ and CD4+/CD45RA+ compared to decidua of healthy pregnant controls. CONCLUSIONS: These changes suggest that deficiency of suppressor activity as well as aberrant immunoregulation exists in decidual tissue of PE women.

Levels of mannose binding lectin in early pregnancy complicated by diabetes mellitus type 1--preliminary report.

The complement system plays an important role as a product of innate and acquired immune reaction. It can be activated via three different routes: the classical pathway, the alternative pathway and the lectin pathway. MBL (mannose-binding lectin) is considered to be a pathogen recognising receptor (PRR), an important factor in recognising pathogen associated molecular pattern (PAMP). The aim of study was to evaluate MBL in early pregnancy of diabetic mothers. Higher values of MBL were observed in diabetic non-pregnant women compared to healthy non-pregnant. Subjects early pregnancy seems to decrease MBL values in both diabetic and healthy pregnant women.

[Peripheral blood neutrophils reactive oxygen intermediates (ROI) production in early pregnancy complicated by diabetes mellitus type I]. / Produkcja reaktywnych form tlenu (RFT) przez neutrofile krwi obwodowej we wczesnej ciazy powiklanej cukrzyca typu 1.

UNLABELLED: The components of maternal innate immune system are activated in pregnancy. Increased number of circulating neutrophils and monocytes, reactive oxygen intermediate production (ROI), increased expression of the adhesion molecules can be observed. The activation status and neutrophils response to stimulators in pregnant diabetic women's comprise, to some extend, both situations pregnancy and diabetes, combined effect of both seems to be very intriguing. AIM OF STUDY: Evaluation production of ROI on peripheral blood neutrophils in early pregnancy of diabetic mothers. MATERIAL: 12 pregnant diabetic women demonstrating good glycemic control and without major complications before pregnancy were studied at 8-12 weeks of first gestation, 11 healthy pregnant women match for weeks of gestation, age and parity; 11 healthy non-diabetic women; 13 diabetic, non-pregnant women, were also studied. METHODS: ROI production was evaluated with chemiluminescence method using standard stimuli fMLP, PMA, OZ and also after 15 min. priming with TNF-alpha. We used MLX Microtiter Plate Luminometr DYNEX, USA. with computer program Revelation 97. For statistic analysis Fisher, Kolmogorow-Smirnow and Wilcoxon tests were used (p < 0.05). RESULTS: 1. Peripheral blood neutrophils of healthy pregnant, diabetic and pregnant diabetic women independently from the stimuli used showed an increase in ROI production 2. Peripheral blood neutrophils change their activity, evaluate by ROI production during pregnancy, diabetes and pregnancy complicated by diabetes. 3. Peripheral blood neutrophils of pregnant diabetes women do not show expected impairment of their function. CONCLUSIONS: Diabetes type 1 with good glycemic control and pregnancy, change the production of ROI by peripheral blood neutrophils but these events do not sum up in diabetic pregnancies.

[The assessment of the effectiveness of the shark liver oil in recurrent aphthous stomatitis treatment: clinical and immunological studies]. / Ocena skutecznosci leczenia aft nawrotowych olejem z watroby rekina w aspekcie badan klinicznych i immunologicznych.

There is no specific and effective treatment for recurrent aphthous stomatitis (RAS), which is connected with its unexplained etiology. The purpose of this study was to determine the clinical efficiency and immunomodulation of the shark liver oil in treatment of recurrent aphthous stomatitis. Twenty-five patients with severe disease received treatment with shark liver oil during a three-month period. The frequency of occurrence of RAS decreased from 1.56 before treatment to 0.95 after treatment and the number of lesions per month was significantly reduced during the third month of treatment and two months after treatment. During two months after treatment 4 patients had no ulcers and an improvement was exhibited in all except 3 of the remaining patients. The treatment had an influence on some immunological parameters. A better response of neutrophils to OZ and PMA were seen. The B cell and T CD3/HLA DR+ cell percentage returned to normal values. A significantly increased percentage of T cells was observed as compared to the before treatment value. The level of C4 and the hemolytic activity of the complement system decreased after treatment and neared the normal values. The shark liver oil contains compounds that have a positive immunomodulation action and alleviate the course of disease.

The interrelationship between production of reactive oxygen intermediates and inflammatory cytokines by peripheral blood leukocytes in vitro from children with food allergy.

In this study, the activity of neutrophils measured using the chemiluminescence test was estimated in 49 children with food allergy. It was shown that average chemiluminescence values without, as well as after, stimulation by formyl-methionyl-leucine phenylalanine and phorbol myristate acetate were considerably higher in children with allergy in comparison to those in the control group. The correlation between the production of reactive oxygen intermediates and interleukin (IL)-4, IL-5 and IL-10 concentration suggests that an autocrine regulatory mechanism may play a role in allergy in children.

[Antiphospholipid autoantibodies in women treated for infertility]. / Wystepowanie autoprzeciwcial antyfosfolipidowych u kobiet leczonych z powodu nieplodnosci.

OBJECTIVE: To evaluate an occurrence of selected antiphospholipid antibodies (APL) in infertility women. STUDY DESIGN: An enzyme-linked immunoabsorbent assay (ELISA) has been used to determine a titre of anticardiolipin antibodies (ACA). APTT (activated partial thromboplastin time) and PT (prothrombin time) measurements and direct commercial assay have been also used to study occurrence of LAC (lupus anticoagulant antibodies). The group of 268 observed women has been divided into five subgroups according to the cause of infertility: I-endometriosis, II-tubal occlusion caused by others diseases than endometriosis, III-occurrence of antisperm antibodies (ASA), IV-polycystic ovariorum syndrome (PCOS) and V-unexplained etiology. Results have been compared to 44 healthy controls. RESULTS: Occurrence of ACA has been found statistically more often in patients suffering from infertility than in controls (17.9% vs 4.5%, p < 0.05). Moreover ACA have been statistically more often observed in patients with endometriosis (21 from 44, 47.7%). ACA were more common in patients with other cause of infertility (except PCOS) compared to controls, though statistically this was not significant. Frequency of ACA was also more common than LAC (17.9% vs 13.6%). Compared to controls LAC tests were more often positive in infertile women with endometriosis (I), tubal occlusion (II) and with the occurrence of antisperm antibodies (III) but results were not statistically significant. CONCLUSIONS: Antiphospholipid antibodies are observed more often in patients suffering from infertility compared to women with a regular fertility. This suggests a role of APL in etiology of infertility. The occurrence of anticardiolipin antibodies in almost every second woman suffering from endometriosis suggests existing of autoimmunologic disturbances in this disease.

[The role of peripheral blood lymphocyte subpopulations in differentiation between preeclampsia and transient hypertension]. / Znaczenie oznaczen subpopulacji limfocytów krwi obwodowej w róznicowaniu pomiedzy preeklampsja a nadcisnieniem przemijajacym w ciazy.

OBJECTIVES: Hypertension is the most frequent complication of pregnancy after 24th week of gestation, occurring in 8% of pregnancies and being the main cause of perinatal mortality and morbidity. It is classified as preeclampsia (PE) or transient hypertension (TH). According to some statements PE and TH are distinct syndromes of different pathogenesis. There are even opinions emphasizing that in most cases TH is in fact undiagnosed chronic hypertension. The role of immunological system in pathogenesis of PE is well known but the hypothesis that immunological events are engaged in pathogenesis of chronic hypertension has not been proved so far. Assuming that TH is closer in its pathogenesis to chronic hypertension than to PE it would be possible to differentiate between TH and PE using some immunological tests. If PE and TH are the same, the differences would be insignificant. DESIGN: The aim of this study was to check the hypothesis that peripheral blood lymphocyte subsets analysis is an useful tool in differentiation between PE and TH and confirmation of their distinct origin. MATERIALS AND METHODS: The study groups consisted of 19 pregnant women with PE (mean age 25.5 +/- 2.5 years, mean gestational age 32.5 +/- 2.5 weeks, 84.2% primiparae) and 14 pregnant women with TH (mean age 27.0 +/- 3.0 years, mean gestational age 33.5 +/- 3.0 weeks, 100% primiparae) diagnosed between 30-37 week of gestation. All women were matched according to gestational age and race. They had no renal diseases or chronic hypertension prior to pregnancy neither had any features of them during the study. Exclusion criteria were: uterine contractions, infection and therapy with steroids before blood sampling. PE and TH were defined according to USA National Health Institute criteria. Peripheral blood was obtained by venipuncture. Standard immunofluorescent marking techniques for whole blood with one-step monoclonal antibodies were performed. Lymphocyte subsets (CD19+, CD3+, CD4+, CD8+, CD3-/CD16+/CD56+, CD3+/CD16+/CD56+, CD8+/CD28+, CD4+/CD45RA+, CD4+/CD45RO+, CD3+/CD69+) analysis was done with flow-cytometer FACSCalibur with 488 nm argon laser. The lymphocyte cells region was chosen with LeucoGATE and analysis performed with SimulSET v.3.1 programme. Statistical analysis was based on Student T test. RESULTS: The differences in peripheral blood lymphocyte subsets composition between PE and TH were insignificant. CONCLUSION: Is that on the basis of peripheral blood lymphocyte subsets analysis PE and TH despite different clinical symptoms seem to have common pathogenesis. However there is possibility that changes observed in peripheral blood are not significantly different in PE and TH because of their low importance for immunopathogenesis.

[The role of cytokines in Th1/Th2 balance in pregnant women with transient hypertension]. / Rola wybranych cytokin w równowadze Th1/Th2 u ciezarnych z nadcisnieniem przemijajacym w ciazy (NP).

OBJECTIVES: There is growing evidence that Th1/Th2 imbalance in cytokine network may play role in immunopathology of preeclampsia (PE). Normal pregnancy is "Th2 phenomenon" while PE is believed to be caused by Th1-shift. There is few data concerning Th1/Th2 imbalance in transient hypertension of pregnancy (TH). DESIGN: The aim of this study was to check the hypothesis that changes in IFN-gamma and IL-2 (Th1 cytokines) levels are accompanied with deficit in TGF-beta (anti-Th1 cytokine) production in TH. MATERIALS AND METHODS: The study group consisted of 10 pregnant women with TH diagnosed between 30-37 week of gestation (mean age 27.0 +/- 3.0 years, mean gestational age 33.5 +/- 3.0 weeks) and 10 women with uncomplicated pregnancy (mean age 26.0 +/- 3.5 years, mean gestational age 35.5 +/- 1.5 weeks). All women were primigravidae matched according to gestational age and race. They had no diabetes mellitus and renal diseases prior to pregnancy neither had any features of them during the study. They had no hypertension before pregnancy. Exclusion criteria were: uterine contractions, infection and therapy with steroids before blood sampling. TH was defined according to USA National Health Institute criteria. Peripheral blood lymphocytes (PBL) were cultured for 72 hours in standard 1640 RPMI medium enriched with 20% FCS, L-glutamine, antibiotics (Sigma) and mitogen phytohemaglutynine (PHA) (Sigma). Cytokine levels were estimated in culture supernatants by using standard ELISA kits according to the indications of the producer (R&D). Statistical analysis was performed with Student-T test. RESULTS: In TH group the levels of IL-2 were higher compared with control group but the differences did not reach statistical significance (4.5 vs. 1.56 pg/ml, t = -1.00, p < 0.1). The same referred to IFN-gamma levels (754.0 vs. 771.0 pg/ml, t = -0.04, NS). However, TGF-beta levels were significantly lower in TH group compared with control group (2459.2 vs. 4156.4 pg/ml, t = -1.47, p < 0.05). Conclusion is that there is a significant deficit of TGF-beta production in peripheral blood lymphocytes of women with TH studied "in vitro". This may contribute to Th1-shift seen in TH similarly to PE.