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Biomedical subjects

H Tchórzewski

Publications and source records attributed to H Tchórzewski.

At least 37 records · Page 2Linked to original sources

[The role of peripheral blood lymphocyte subpopulations in differentiation between preeclampsia and transient hypertension].

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.

Adult↗

[Evaluation of some neutrophil adhesion molecules in pregnancy complicated by diabetes mellitus type 1].

UNLABELLED: The effective functions of human peripheral blood neutrophil are modulated by regulatory molecules depending on receptor agonists and adhesion molecules. OBJECTIVES: Evaluation of percentage of neutrophil adhesion molecules in pregnancy of diabetic mothers as well as their expression on peripheral blood neutrophils. MATERIAL: A-12 pregnant diabetic women at 7-13 and 34-36 weeks of first gestation, B-11 healthy pregnant women match for gestation, age and parity; C-11 healthy non-diabetic women; D-14 diabetic, non-pregnant women. METHODS: MFI expression and percentage of CD18, CD11b, CD62L and CD54 were evaluated resting and activated by 30 min., fMLP preincubation using monoclonal antibodies anti-CD11b and anti-CD62L, employing Flow Cytometer FACSCalibur with computer program CELLquest. RESULTS: Expression of CD11b and CD18 receptor on leukocyte of A, B, D groups was higher spontaneous and after stimulation by fMLP in I-st trimester compare to group C and then lowered in IIIrd. The significant difference in CD62L percentage was noticed resting and activated neutrophils in group A, which was lower compared to C and expression of CD62L on peripheral blood neutrophils was higher in group B compared to other studied groups in Ist trimester. There was an increase in expression of CD54 on resting neutrophils of group D when compared with C. A significant difference between Ist and IIIrd trimester was noticed in group A in expression of CD62L,CD62L/fMLP,CD54 on peripheral blood neutrophils, which were higher in the third trimester. Expression of CD62L was higher in the group A when compared to B. CONCLUSIONS: Diabetes type 1 and pregnancy changed the percentage and MFI of neutrophil adhesion molecules.

Adult↗

[Effect of maternal labor and mode of delivery on function of neonatal cord blood neutrophils].

There are data to suggest that the method of delivery modifies the immunological balance of newborn infants. In this study, an effect of maternal labor and mode of delivery on the neutrophil activity was evaluated in healthy full-term neonates. Neutrophils were obtained from cord blood of two group neonates: 14 vaginally delivered and 12 delivered by elective cesarean section without preceding labor. Neutrophil functions were studied by measuring the chemotaxis, chemiluminescence and phagocytosis activity and by investigating the expression of phagocyte receptors CD11b on neutrophils using receptor-specificMoAbs and immunofluorescence flow cytometry. Infants born by elective cesarean section had significantly higher chemiluminescence activity, chemotaxis and higher expression of CD11b receptors than those delivered vaginally. We suggest that physiological changes associated with stress during labor could explain these differences observed in neutrophil functions.

Cesarean Section↗

[The influence of paternal lymphocyte immunization on the balance of Th1/Th2 type reactivity in women with unexplained recurrent spontaneous abortion].

OBJECTIVES: Paternal lymphocyte immunization has been proposed as an efficient treatment for unexplained recurrent spontaneous abortion (RSA), however precise mechanism that underlie the benefits of this immunotherapy are still unclear. It was proposed that successful pregnancy is reminiscent of T helper 2 (Th2)--dominant situation but unsuccessful pregnancy is a Th1-type situation. The aim of the study was the evaluation of influence of paternal lymphocyte immunization on the balance of Th1/Th2--type reactivity in women with unexplained recurrent spontaneous abortion. MATERIAL AND METHODS: 8 patients with a history of 3 or more consecutive primary spontaneous abortions of unknown etiology and no positive autoimmune factors were selected for the study. Immunization with paternal lymphocytes, obtained from 100 ml of peripheral blood, was performed twice prior conception with a 4-week interval. The following immunological parameters were studied: peripheral blood T lymphocyte subpopulations (CD3, CD4, CD8) and secretion of the Th1-type cytokines (IL-2, IFN-gamma), Th2-type cytokines (IL-6) and TGF-beta 1 by phytohaemaglutinin-stimulated peripheral blood lymphocytes. Evaluation of immunity parameters were performed before and 2 weeks after immunotherapy. RESULTS: It was found that paternal lymphocytes immunization significantly increase the percentage of CD4 T lymphocytes (37.11 +/- 7.65 vs. 41.38 +/- 5.57, p = 0.007). Immunotherapy also leads to a significant enhancement in Th2-type cytokines (IL-6) secretion (22,677 +/- 17,907 mg/ml vs. 44,550 +/- 15,907 mg/ml, p = 0.008) and a significant decreasing in Th1-type cytokines (IL-2) secretion (6.50 +/- 5.98 mg/ml vs. 0.00 +/- 0.00 mg/ml, p = 0.0179). CONCLUSIONS: The data of the present studies suggest that paternal lymphocytes immunization modulate of immunity in women with unexplained recurrent spontaneous abortion. Our studies indicate a shift in the balance fo cytokine profiles away from Th1-type reactivity to a Th2-type reactivity after immunotherapy.

Abortion, Habitual↗

[Antiphospholipid autoantibodies in women treated for infertility].

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.

Adult↗

Priming effect of met-enkephalin and beta-endorphin on chemiluminescence, chemotaxis and CD11b molecule expression on human neutrophils in vitro.

The opioid peptides are widely distributed throughout the body, and they are generated during stress and inflammatory reaction. Opioids are involved in the communication between the immune and neuroendocrine systems. In the present study we have investigated the ability of both met-enkephalin and beta-endorphin to stimulate and prime the human neutrophils for enhanced chemiluminescence (CL) and chemotaxis induced with fMLP, OZ or PMA. We have also tested the effect of beta-endorphin and met-enkephalin on CD11a, CD11b, CD18 and CD16 molecule expression on PMN in vitro. PMN from ten healthy donors were incubated in vitro with different concentrations of beta-endorphin or met-enkephalin, and the CL response was evaluated with luminometer. To assess the effect of opioid peptides on CD11a, CD11b, CD18 and CD16 molecule expression the whole blood samples were incubated with different concentrations of the opioids, then the white cells were labelled with respective PE-conjugated MoAb and evaluated by flow cytometry. We have shown that: (1) met-enkephalin and beta-endorphin at physiological concentrations relevant to that of in vivo (10(-8) and 10(-6) M) enhanced fMLP, PMA or OZ stimulated chemiluminescence and induced chemotactic response, (2) High concentrations of beta-endorphin (10(-3) M) or met-enkephalin (10(-5) M) decreased the CL response of PMN in vitro, (3) The opioid peptides at lower concentrations resulted in CD11b and CD18 molecule up-regulation on neutrophils. We may conclude that opioid peptides in physiological concentration are involved in neutrophil priming whereas in higher concentration exert immunosuppressive potency. Opioid peptides like inflammatory cytokines may prime the neutrophils inflammatory response.

Animals↗

The immunoregulatory abilities of polymorphonuclear neutrophils in the course of multiple sclerosis.

The polymorphonuclear neutrophils (PMN) possess sufficient potential to affect both immune response and inflammation, however it has not been yet described in the course of multiple sclerosis (MS). We have studied binding of fluorescein isothiocyanate (FITC)- stained TNF-alpha by PMN, the expression of CD11a, CD11b, and CD18 molecules of beta2-integrines and the expression of CD10 (neutral endopeptidase-NEP) and of CD13 (aminopeptidase N; APN) antigens on PMN in three different groups of MS patients. The control group included neurological patients (OND) with noninflammatory diseases. The obtained results have proved that during MS exacerbation and in the course of chronic progressive MS, PMN reveal several forms of preactivation, including significantly higher stained-TNF-alpha binding, higher expression of CD11b and CD18, as well as CD10 and CD13 antigens, in comparison with MS remission or OND. We suggest that the increased expression of these molecules on PMN of MS patients in exacerbation of the disease and to a lower degree in the course of CP-MS is a result of PMN priming, and directly prove the PMN involvement in the disease pathogenesis.

Adult↗

The comparison of some biological activities of lipopolysaccharides obtained from smooth and rough Proteus mirabilis strains.

The biological activity of lipopolysaccharide (LPS) obtained from Proteus mirabilis smooth and rough strains was investigated. The tested endotoxins (differing in polysaccharide chain lenght) were isolated from wild S1959 strain as well as from its rough mutants Ra and Re. Induction of tumor necrosis factor-alpha (TNF-alpha), and nitric oxide production as well as activation of complement system by lipopolysaccharide are the pathophysiological reaction in a host response to gram-negative bacteria. In this study, it was found that S (S1959), Ra (R110) and Re (R45) chemotypes of LPS similarly induced the human neutrophils to release TNF-alpha. In contrast none of the LPS stimulated the neutrophils to synthesis of nitric oxide regardless of doses used and culture time. Te Re form of LPS showed the strongest anticomplementary activity.

Animals↗

Listeria monocytogenes infection in mice treated with pentoxifylline.

The course of L. monocytogenes infection was followed in mice treated with pentoxifylline (POF), a known inhibitor of endogenous tumor necrosis factor (TNF) formation. Administration of POF caused a delay in L. monocytogenes elimination which was probably related to a reduction the listericidal activity of macrophages and to an attenuation of delayed type hypersensitivity (DTH) to Listeria antigens. In spite of this, some POF-treated mice were protected from lethal effects of virulent L. monocytogenes bacteria.

Animals↗

Role of tyrosine kinase enzymes in TNF-alpha and IL-1 induced expression of ICAM-1 and VCAM-1 on human umbilical vein endothelial cells.

The aim of this study was to analyse the potential roles of protein kinase enzymes in tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) induced expression of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on human umbilical vein endothelial cells (HUVEC). The authors observed a marked increase in ICAM-1 and VCAM-1 expression on HUVEC stimulated for 24 h by TNF-alpha (10 ng/ml) or IL-1 (20 ng/ml). Pre-treatment of HUVEC for 30 min with protein tyrosine kinase (PTK) inhibitors genistein and herbimycin A (10 micrograms/ml and 0.5 microgram/ml, respectively) before stimulation with IL-1 did not affect the expression of these molecules. Similar results were observed with respect to VCAM-1 expression on HUVEC stimulated by TNF-alpha. In contrast, pre-incubation of HUVEC with PTK inhibitors prior to the addition of TNF-alpha significantly enhanced subsequent expression of ICAM-1, although spontaneous expression of ICAM-1 on unstimulated HUVEC was unaffected. Western blot analysis demonstrated a significant increase in phosphorylated tyrosine protein levels in HUVEC stimulated by TNF-alpha, and significantly lower levels of these proteins in TNF-alpha stimulated HUVEC pre-treated with PTK inhibitors. These results demonstrate that IL-1 induced ICAM-1 and VCAM-1 expression does not result from activation of PTK-dependent pathways. In the case of TNF-alpha induced responses, the selective co-stimulatory effect of this cytokine in combination with PTK inhibitors on ICAM-1 expression suggests a complicated intracellular pathway of TNF-alpha induced ICAM-1 expression, possibly involving down-modulation of increases in ICAM-1 by PTK enzymes.

Benzoquinones↗

Myoinositol may be a factor in uremic immune deficiency.

In 10 chronic uremic patients on regular hemodialysis treatment and 10 healthy subjects in vitro PHA-induced peripheral blood mononuclear cell (PBMNC) and T-cell enriched lymphocyte proliferative responses were found to be impaired in the presence of myoinositol in the concentration generally observed in the blood serum of chronic uremic patients on regular hemodialysis treatment (600 mumol/l), while it remained unchanged in the presence of myoinositol in the concentration observed in normal blood serum (30 mumol/l). However, both myoinositol concentrations did not affect PMA-induced PBMNC and T-cell enriched lymphocyte proliferative responses, which suggests that inhibitory effect of the high myoinositol concentration on PHA-induced immune cell proliferation is cell membrane-related. In addition, myoinositol (600 mumol/l) significantly depressed CD3, CD4 and HLA-DR antigen expression on PHA-activated PBMNC surface in chronic uremic patients and healthy subjects, while CD8 antigen expression remained unaffected. The results seem to indicate that myoinositol, in the concentrations observed in uremic blood serum, may possibly share the responsibility for uremic immune deficiency.

Adult↗

CD3-/HLA-DR+, CD3+/HLA-DR+ cell levels and PHA stimulation of lymphocytes obtained from women with recurrent spontaneous abortions and subjected to partner's lymphocyte immunization.

It is assumed that 80% of recurrent spontaneous abortions (RSA) of unknown etiology are of immunological origin. The proposed experimental treatment of those patients could be immunization with paternal lymphocytes, though the exact mechanism through which this therapy exerts its effect is still unknown. Some authors suggest that lymphocyte alloimmunization alters the number and activity of particular subpopulations of peripheral blood lymphocytes. The aim of this study was to evaluate the proliferative O activity of lymphocytes stimulated with phytohemagglutinin (PHA) and the percentage of peripheral lymphocytes carrying CD3+/HLA-DR+ and CD3-/HLA-DR+ markers, present in 32 selected for this study RSA women subjected to paternal lymphocytes' alloimmunization performed prior to conception and in early pregnancy. We conclude that, the post-immunization changes seem to have different quantitative and qualitative character comparing to those seen in normal pregnancy. They seem to avert unfavourable immunological phenomena observed in RSA women, allowing for successful continuation of pregnancy.

Abortion, Habitual↗

[Cytometric analysis of lymphocyte phenotype in cord and newborn blood].

At birth and in early childhood, phenotypic and functional investigation of the immune system is often necessary to identify primary immunological disorders and may provide additional diagnostic and prognostic information in certain intrauterine infections. We compared, using of flow cytometric immunophenotyping, a broad panel of lymphocyte subsets in paired samples obtained from healthy infants at birth (cord blood) and at 3 days of age. Results were compared to those of normal children (7 to 17 years) evaluated using the same methods. Significant age-related differences were demonstrated in lymphocyte subpopulations. Our data can be used as age-matched reference values for blood lymphocyte immunophenotyping.

Adolescent↗

The role of receptors for tumour necrosis factor-alpha in the induction of human polymorphonuclear neutrophil chemiluminescence.

Tumour necrosis factor-alpha (TNF-alpha) is a potent mediator of inflammation, which exerts profound effects on polymorphonuclear neutrophils (PMN). TNF-alpha binds to distinct cell surface receptors termed p55 and p75, expressed in approximately equal amounts on the PMN surface. We have studied the effects of TNF-alpha on the priming of F-Met-Leu-Phe (FMLP)-stimulated oxidative metabolism of PMN, using a luminol-enhanced chemiluminescence assay, and have examined the relative roles of PMN receptors for TNF-alpha in priming this oxidative metabolism, using antibodies with p55 and p75 receptor-specific agonistic and antagonistic activities. We have obtained the following results: (1) Antibody Htr-9 with agonistic activity at the p55 receptor mimicked the effect of TNF-alpha; however, a combination of Htr-9 and TNF-alpha did not results in any further increase in chemiluminescence relative to the response observed with TNF-alpha alone. The p75 agonistic antibody MR2-1 actually decreased basal and FMLP-enhanced chemiluminescence. Additionally, MR2-1 substantially inhibited the effects of both TNF-alpha itself and of the p55 agonist Htr-9. (2) Addition of antibodies with antagonistic activities at the p55 (antibody TBP-2) and p75 (antibody Utr-1) receptors resulted in a marked inhibition of the PMN response to TNF-alpha. A combination of both Utr-1 and TBP-2 was most effective at inhibiting the action of TNF. We have confirmed previously published observations that TNF-alpha alone effectively stimulates the oxidative metabolism of PMN in vitro, and that pre-incubation of PMN with TNF-alpha enhances subsequent generation of oxidative metabolites in response to FMLP. We conclude that both p55 and p75 receptors play a critical role in mediating the activation of PMN by TNF-alpha.

Adult↗

Oxidative burst response of neutrophils primed with PreS1 antigen of hepatitis B virus in patients with chronic hepatitis B and convalescents.

The pathogenesis of liver damage in the course of hepatitis B virus (HBV) infection depends on the host's specific and non-specific immune response to various viral antigens. The role of polymorphonuclear neutrophils (PMN) in the natural immune reaction and during secondary microbial infections is well documented. Increased free radical production is associated with many pathological conditions such as shock, ischaemia or chronic inflammatory diseases. We studied the oxidative metabolism of neutrophils in patients with chronic HBV and after recovery (convalescents). The effect of the PreS1 fragment of HBV antigen on some neutrophil functions in vitro was also examined. There were significant differences in the values of spontaneous and stimulated oxidative burst of neutrophils, measured using luminol-chemiluminescence, in patients with HBV when compared with the convalescents. PreS1 antigen did not by itself induce the respiratory burst in human neutrophils but it potentiated their response to a second stimulus. Hence we observed a priming of neutrophils, for an enhanced respiratory burst, by PreS1 antigen.

Adult↗

Protease inhibitors diminish lymphocyte stimulation in vitro.

Lymphocyte stimulation initiates activation of signal transduction pathways presumably enzymatic in nature. The next step includes gene activation and specific protein molecule production. Proteases of different specificity are crucial in enzyme activation: protein cleavage and biologically active molecule production. To elucidate the role of proteases in peripheral blood mononuclear cell (PBMC) activation, the broad specificity inhibitors of thiol (PHMB) and serine (TLCK) proteases have been used in vitro. Both inhibitors diminished the PHA-induced lymphocyte stimulation when applied at the beginning of culture; inhibitory effect was abrogated when inhibitors were introduced to the culture system 4 h later. Exogenous IL-2 abolished inhibition. PHMB activity was reversible whereas TLCK was irreversible. The inhibition of T lymphocyte-enriched proliferation is more distinctive as compared to that of PBMC. It can be concluded that proteases are involved in early events of lymphocyte proliferation and IL-2 production which is responsible for different stages of cell growth.

Cells, Cultured↗

Effect of pre-S1 antigen on human lymphocyte proliferative responses.

The peripheral blood mononuclear cells (PBMCs) from patients acutely infected with HBV and recovered completely (n = 20), patients with chronic hepatitis B infection (CHB)- (n = 10) and HBsAg-positive carriers (n = 9) and healthy individuals (n = 8) were studied for their in vitro proliferative response to a synthetic pre-S1(20-49)x4 antigen. PBMCs from convalescents showed significant proliferative response in the presence of synthetic pre-S1 antigen. PBMCs from CHB- and HBsAg-positive exhibited reduced proliferative response not only to Pre-S1 antigen but also to nonspecific mitogens. This study suggests that the immune recognition of pre-S1 antigen and response of PBMCs to the pre-S1 antigen may be an important part of the normal human response to HBV infection. Failure to clear the HBV infection with development of the chronic carrier state may be caused by the lack of an efficient pre-S1 antigen-specific response.

Adult↗