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1.
Artigo em Inglês | MEDLINE | ID: mdl-19274928

RESUMO

BACKGROUND: The influence of food allergens on the profile of inflammatory markers in children with asthma has not been investigated. To ascertain the influence of food allergens on the intensity of the inflammatory process, a cytokine profile was determined before and after a food challenge test in the peripheral blood of children with asthma and coexistent food allergy. MATERIAL AND METHODS: We studied 22 children with asthma and immunoglobulin (Ig) E-dependent food allergy. Oral challenge tests were carried out using double-blind placebo-controlled food challenge (DBPCFC). Blood was sampled before, and 4 and 24 hours after the oral challenge test. The inflammatory markers interleukin (IL) 4, IL-5, IL-10, tumor necrosis factor (TNF) a, intereron (IFN)-gamma, sIL-2R, and sCD23 were evaluated. The level of cytokines in serum was determined using a commercial enzyme-linked immunoassay Bender Med Systems (Vienna, Austria). RESULTS: The median IL-4 level before the challenge test was 23.5 pg/mL, after 4 hours it was 38.8 pg/mL, and after 24 hours it was 35.4 pg/mL. The median IL-5 levels measured at the same time points were 4.6 pg/mL, 5.7 pg/mL, and 7.5 pg/mL. A significant increase in IL-4 and IL-5 levels 4 hours (P = .0006; P = .006) and 24 hours (P = .014; P = .015) after food challenge was observed. No statistically significant differences in the levels of the other cytokines during allergen or placebo challenge tests were recorded. CONCLUSIONS: Determination of plasma IL-4 and IL-5 levels can be a useful tool for evaluation of the effects of food challenge tests on children with asthma and coexisting IgE-dependent food allergy. The results of determining serum IL-10, TNF-alpha, interleukin (IL) IFN-gamma, sIL-2R, and sCD23 levels during the challenge test are not significant.


Assuntos
Asma/imunologia , Citocinas/sangue , Hipersensibilidade Alimentar/imunologia , Adolescente , Criança , Método Duplo-Cego , Ensaio de Imunoadsorção Enzimática , Hipersensibilidade Alimentar/diagnóstico , Humanos , Testes Imunológicos/métodos , Estatísticas não Paramétricas
2.
Scand J Immunol ; 68(2): 145-52, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18702744

RESUMO

The study was aimed at evaluating the involvement of sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 and reactive oxygen species (ROS) in systemic inflammatory response syndrome (SIRS) development in severely burned children and at assessing the prognostic value of the immunological markers studied. The study comprised 37 patients (17 burned children and 20 controls). Serum levels of the markers determined by means of ELISA and respiratory burst of neutrophils as well as p55 and p75 tumour necrosis factor-alpha (TNF-alpha) receptor expression using flow cytometry were evaluated twice. The burned children presented significantly higher levels of IL-10 and cytokine inhibitors within the first 6-24 h after injury compared with controls (P < 0.05). The decreased oxygen metabolism of neutrophils and increased TNF-alpha receptor expression were found on admission. Moreover, a significant decrease in initially high sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 concentrations (P < 0.05) and reduced expression of TNF-alpha receptors (P < 0.05) were observed after burn therapy, whereas ROS generation evidently augmented (P < 0.05). Four of our children who developed hypovolaemic shock revealed a significantly lower ROS generation and higher concentrations of soluble TNF-alpha receptors and IL-1 ra together with IL-10, IL-13 compared with children with good outcome (P < 0.05). Our results revealed the involvement of both ROS, soluble TNF-alpha receptors and IL-1 ra in the development of SIRS in burned children; their monitoring allows for an assessment of the systemic inflammatory reaction activity. The neutrophil BURSTTEST and IL-1 ra might have been clinically helpful markers of SIRS prognosis.


Assuntos
Biomarcadores/sangue , Queimaduras/sangue , Citocinas/sangue , Neutrófilos/metabolismo , Síndrome de Resposta Inflamatória Sistêmica/sangue , Queimaduras/complicações , Queimaduras/imunologia , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Lactente , Inflamação/sangue , Inflamação/etiologia , Inflamação/imunologia , Interleucina-10 , Interleucina-13 , Masculino , Prognóstico , Espécies Reativas de Oxigênio/metabolismo , Receptores do Fator de Necrose Tumoral/sangue , Síndrome de Resposta Inflamatória Sistêmica/etiologia , Síndrome de Resposta Inflamatória Sistêmica/fisiopatologia
3.
Clin Exp Immunol ; 127(1): 78-84, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11882036

RESUMO

CD30 is expressed on activated T cells that, as has been suggested, preferentially produce IFN-gamma. Interleukin 12 increases antigen-induced CD30 expression on T cells and IFN-gamma production. Synthesis of IFN-gamma can be augmented further by IL-18. The aim of our study was to investigate whether IL-18 affects the IL-12 induced CD30 expression and cytokine production by allergen or PPD specific T cells. Mononuclear cells of healthy or atopic volunteers were stimulated with PPD or allergen, respectively, to obtain specific T cell lines. T cells were restimulated with appropriate antigen and antigen-presenting cells in the presence of IL-12, IL-18 or a combination of these cytokines. After 3 days, expression of CD30 was investigated on CD4 and CD8 T cells and IFN-gamma and IL-4 cytokine production was estimated in the culture supernatants. Flow cytometric analyses showed no effect of IL-18 on CD30 expression during IL-12 co-stimulation. At the same time after the optimal stimulation for CD30 expression, the levels of IFN-gamma were high in PPD-stimulated cell lines but have not been up-regulated by IL-18. IFN-gamma levels were much lower in allergen-stimulated T cells and although they were up-regulated by IL-12 there was no additional or synergistic effect from IL-18. IL-18, however, increased production of IL-4 in allergen-stimulated cell lines. Our studies provide new information about IL-18 activity on human cells and question its exclusive role in Th1 mediated responses.


Assuntos
Interferon gama/imunologia , Interleucina-12/imunologia , Interleucina-18/imunologia , Interleucina-4/imunologia , Antígeno Ki-1/imunologia , Subpopulações de Linfócitos T/imunologia , Alérgenos/imunologia , Células Cultivadas , Citometria de Fluxo , Humanos , Interferon gama/metabolismo , Interleucina-12/farmacologia , Interleucina-18/farmacologia , Interleucina-4/metabolismo , Antígeno Ki-1/biossíntese , Ativação Linfocitária/imunologia , Neutrófilos/imunologia , Tuberculina/imunologia
4.
Int J Cardiol ; 79(1): 49-59, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11399341

RESUMO

Abnormal platelet function has been hypothesised to play a role in the haemostatic abnormalities in cyanotic congenital heart disease (CCHD) patients. Using whole blood flow cytometry we found that platelets from cyanotic patients were hyperreactive and we related such hyperreactivity directly to young age, unoperated state, high haematocrit, reduced saturation with oxygen and low platelet count. Circulating platelets from CCHD children showed significantly enhanced P-selectin expression (P<0.004) and remained more reactive to 0.2 IU/ml thrombin, 1-8 microM TRAP and 2-4 microM ADP (P<0.04), especially in younger (0-3-year-olds) patients. Such a platelet 'priming' largely concerned CCHD children who were not subjected to modified Blalock-Taussig shunts in the past (non-MBTS). Only non-MBTS cyanotic children, but not MBTS-operated patients, showed significantly higher platelet reactivity compared to controls in response to ADP or 1 microM TRAP with respect to P-selectin expression (p<0.05) and in response to all examined agonists with respect to GPIb expression (P<0.045). The enhanced P-selection expression in MBTS-operated CCHD children and reduced GPIb expression in non-MBTS patients, especially in younger patients, were positively associated with the occurrence of the polymorphic variant Pl(A2) of platelet membrane glycoprotein IIIa gene. Altered blood morphology parameters (elevated RBC, Hb, Hct and MCHC, for all P<0.0005) in CCHD children correlated with the enhanced degranulation of circulating blood platelets and their hyperreactivity in response to some agonists (P<0.05). Overall, our data encourage the reasoning that circulating platelets are remarkably hyperreactive in non-MBTS cyanotic children, which are at higher risk to often encounter platelets activation in circulation. It seems unlikely that the apparently unchanged platelet reactivity in MBTS-operated children is due to the advantageous effects of the shunt, since these patients showed neither altered haematological parameters nor improved oxygen carrying capacity. Otherwise, it may rather result from more frequent episodes of platelet degranulation and preactivation in the past, and/or post-operative enhanced platelet consumption.


Assuntos
Prótese Vascular , Cardiopatias Congênitas/sangue , Ativação Plaquetária/fisiologia , Criança , Pré-Escolar , Feminino , Citometria de Fluxo , Cardiopatias Congênitas/genética , Cardiopatias Congênitas/fisiopatologia , Humanos , Lactente , Masculino , Fragmentos de Peptídeos/farmacologia , Ativação Plaquetária/efeitos dos fármacos , Polimorfismo Genético/genética
5.
Arch Immunol Ther Exp (Warsz) ; 49(2): 155-61, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11348020

RESUMO

This study was to evaluate the levels of the proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha) and the cytokine inhibitors soluble TNF-alpha receptor (sTNFR) and interleukin (IL-1) receptor antagonist (IL-1ra), as well as the intensity of oxidative metabolism of peripheral blood polymorphonuclear leukocytes in the course of sepsis in newborns. An increase of TNF-alpha, sTNFR and IL-1ra concentrations was found in the blood serum of the patients at the time of diagnosis. This was further accompanied by polymorphonuclear leukocyte stimulation and, as a consequence of prolonged bacterial antigen stimulation, functional exhaustion of these cells and their diminished oxidative metabolism was observed. Within the same time period, an enhanced expression of p55 and p75 TNF-alpha receptors on polymorphonuclear leukocyte cell surfaces was found. It was indicated that the applied pharmacotherapy caused a decrease of the initially elevated concentrations of TNF-alpha and proinflammatory cytokine inhibitors (sTNFR, IL-1ra). The intensive therapy of sepsis was associated with the increased oxidative burst of polymorphonuclear leukocytes along with the decrease of p55 and p75 expression on their cell surfaces.


Assuntos
Citocinas/antagonistas & inibidores , Citocinas/sangue , Neutrófilos/imunologia , Neutrófilos/metabolismo , Sepse/sangue , Sepse/etiologia , Antígenos CD/sangue , Membrana Celular/imunologia , Membrana Celular/metabolismo , Humanos , Lactente , Recém-Nascido , Proteína Antagonista do Receptor de Interleucina 1 , Receptores do Fator de Necrose Tumoral/sangue , Receptores Tipo I de Fatores de Necrose Tumoral , Receptores Tipo II do Fator de Necrose Tumoral , Explosão Respiratória , Sepse/imunologia , Sialoglicoproteínas/sangue , Solubilidade , Fator de Necrose Tumoral alfa/metabolismo
6.
Platelets ; 12(2): 99-107, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11297038

RESUMO

Neonatal platelets have been occasionally reported to show a reduced response to various agonists. The molecular mechanism(s) of such a depressed reactivity remains unclear. To further address this problem we studied neonatal platelet activation with thrombin, TRAP (thrombin receptor activating peptide, Ser-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe) and ADP in 42 healthy 1-2 day old neonates using a whole peripheral blood flow cytometry. The neonates did not show an increased fraction of P-selectin-positive circulating platelets, whereas the expression of GPIb (glycoprotein Ib) in resting neonatal platelets was significantly lower compared to adults. Neonatal platelets were significantly less reactive than adult platelets to thrombin and TRAP, especially at lower agonist concentrations, but not to ADP or when incubated for 1 h at room temperature. Activation of neonatal platelets with agonists resulted in a marked alterations in the expression of P-selectin, whereas the internalization of GPIb was not affected. The reduced neonatal platelet sensitivity to thrombin and TRAP was accompanied by significantly reduced ATIII (antithrombin III) and increased prothrombin fragment F(1+2) in neonatal plasma. We conclude that various receptor systems potentially able to bind thrombin are relatively insensitive in neonatal platelets. The novelty of our work is that neonatal platelet hyposensitivity is not a generalized phenomenon, but concerns only selected agonists and selected receptor systems.


Assuntos
Plaquetas/efeitos dos fármacos , Recém-Nascido/sangue , Ativação Plaquetária/efeitos dos fármacos , Receptores de Trombina/agonistas , Difosfato de Adenosina/farmacologia , Adulto , Fatores Etários , Antitrombina III/análise , Plaquetas/química , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Selectina-P/análise , Fragmentos de Peptídeos/análise , Peptídeo Hidrolases/análise , Complexo Glicoproteico GPIb-IX de Plaquetas/análise , Proteínas/farmacologia , Protrombina/análise , Receptor PAR-1 , Trombina/farmacologia
7.
Pol Merkur Lekarski ; 8(46): 214-5, 2000 Apr.
Artigo em Polonês | MEDLINE | ID: mdl-10897617

RESUMO

It has been suggested that the immune system is involved in the development of lipoid nephrosis. The aim of the study was to estimate whether an elevated serum concentration of soluble interleukin-2 receptor (sIL-2R) is caused by increased expression of IL-2 receptors on the surface of lymphocytes T (CD3+CD25+). 20 children with lipoid nephrosis and 15 healthy children served as control were evaluated. T cell subpopulations were assayed with flow cytometry, using monoclonal antibodies. The concentration of sIL-2R was determined by enzyme-linked immunosorbent assay (ELISA). The patients with lipoid nephrosis had elevated serum levels of sIL-2R compared with healthy controls. Although the subpopulation of T cell CD3+CD25+ was decreased in nephrotic children. These results could testify against direct contribution of T cell immune response in the pathogenesis of lipoid nephrosis.


Assuntos
Complexo CD3/sangue , Nefrose Lipoide/sangue , Receptores de Interleucina-2/sangue , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino
8.
Med Sci Monit ; 6(4): 722-8, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11208399

RESUMO

INTRODUCTION: Excessive blood loss, as a result of augmented postoperative drainage, is considered one of the most serious cardiosurgical complications. The compounding constitutive anemia seems particularly harmful for patients with coronary artery disease. Aprotinin (Trasylol), a non-specific serine protease inhibitor, is successfully used to reduce excessive postoperative bleeding in such patients. The aim of our study was to verify the hypothesis whether aprotinin used during cardiopulmonary bypass procedure affects hemostatic parameters, which might be crucial for the elevated risk of thromboembolic complications. MATERIAL AND METHODS: The group of 54 patients subjected to coronary artery surgical treatment included 30 patients, who were given intraoperatively 3 million KIU aprotinin each, and 24 subjects non-treated with aprotinin. Aliquots of blood were withdrawn at several time intervals, until the 5th day after the operation. Whole blood platelet activation and reactivity (the expressions of P-selectin and glycoprotein Ib) were monitored by means of flow cytometry. In addition, several plasma parameters, like PAI-1, t-PA, D-dimers, prothrombin fragment F1 + 2, fibrinogen, ATIII activity, troponin I and CK-MB, as well as platelet count were determined at each time point. RESULTS: In this study we confirmed the essential advantage of the use of aprotinin: both the postoperative blood drainage and the blood units to be transfused postoperatively to cardiosurgical patients were vastly reduced in the aprotinin-treated subjects. The enhanced overall frequency of perioperative myocardial infarction events was not attributed to this group of patients, nor the non Q-wave infarctions were observed more often in patients treated with aprotinin. In these patients, fibrinolysis parameters tended to be depressed (with increased PAI-1 dominating over elevated t-PA) on the first day after the operation, and no significant differences with regard to fibrinogen, prothrombin fragment F1 + 2, troponin I and platelet count. There was a continuous rise in D-dimers in all the postoperative patients, which lasted until the third day and tended to reach plateau at the 5th day after the operation. We failed to reveal the preventive effects of aprotinin on platelet function: both platelet activation and reactivity remained apparently unchanged. Overall, our results rather support the reasoning on the advantageous effects of low doses of aprotinin. The use of this inhibitor reduces the risk of postoperative undesirable bleeding and results in a decreased postoperative drainage and reduced transfused blood units. On the other hand, however, a higher incidence of perioperative Q-wave infarction in the aprotinin-treated patients, although purely apparent and not statistically significant, might question the unlimited safety of the use of aprotinin in cardiovascular operations.


Assuntos
Aprotinina/administração & dosagem , Aprotinina/efeitos adversos , Ponte de Artéria Coronária/efeitos adversos , Ponte de Artéria Coronária/métodos , Hemostáticos/administração & dosagem , Hemostáticos/efeitos adversos , Idoso , Coagulação Sanguínea/efeitos dos fármacos , Feminino , Fibrinólise/efeitos dos fármacos , Hemostasia/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Ativação Plaquetária/efeitos dos fármacos , Hemorragia Pós-Operatória/prevenção & controle , Inibidores de Serina Proteinase/administração & dosagem , Inibidores de Serina Proteinase/efeitos adversos , Tromboembolia/etiologia
9.
Exp Cell Res ; 245(2): 303-12, 1998 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-9851871

RESUMO

Curcumin is a well-known natural compound with antiinflammatory properties. Its antiproliferative effect and ability to modulate apoptotic response are considered essential in cancer therapy. The physicochemical properties of curcumin suggest membranous localization, which prompted an investigation of the mechanisms of membrane disturbances evoked by curcumin. We chose the erythrocyte as a convenient model for studying membrane effects of curcumin and showed its nonspecific, apoptosis-independent way of action. Curcumin was found to expand the cell membrane, inducing echinocytosis. Changes in cell shape were accompanied by transient exposure of phosphatidylserine. Membrane asymmetry was recovered by the action of aminophospholipid translocase, which remained active in the presence of curcumin. Lipids rearrangements and drug partitioning caused changes of lipid fluidity. Such nonspecific effects of curcumin on cellular membranes would produce artifacts of apoptosis measurement, since several methods are based on membrane changes.


Assuntos
Apoptose , Curcumina/farmacologia , Membrana Eritrocítica/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Proteínas de Transferência de Fosfolipídeos , Adulto , Anti-Inflamatórios não Esteroides/farmacologia , Antineoplásicos/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Proteínas de Transporte/metabolismo , Tamanho Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Membrana Eritrocítica/metabolismo , Eritrócitos/citologia , Eritrócitos/metabolismo , Citometria de Fluxo , Humanos , Metabolismo dos Lipídeos , Fluidez de Membrana/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Fragilidade Osmótica/efeitos dos fármacos , Fosfatidilserinas/farmacologia , Temperatura , Fatores de Tempo
10.
FEBS Lett ; 433(3): 287-93, 1998 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-9744813

RESUMO

Curcumin (diferuoylmethane) is a natural compound with anticarcinogenic activities which is able to exert either proapoptotic or antiapoptotic effects in different cell types. This paper focuses on the sequence and extent of primary events induced by curcumin, in comparison with those occurring during dexamethasone-induced apoptosis in rat thymocytes. It also presents annexin VI-FITC as a new probe for studying membrane asymmetry. Curcumin readily penetrates into the cytoplasm, and is able to accumulate in membranous structures such as plasma membrane, endoplasmic reticulum and nuclear envelope. Curcumin-treated cells exhibit typical features of apoptotic cell death, including shrinkage, transient phosphatidylserine exposure, increased membrane permeability and decrease in mitochondrial membrane potential. However, nuclei morphology, DNA fragmentation, the extent and time-course of membrane changes are different from those observed during dexamethasone-induced apoptosis, suggesting that, despite many similarities, the mode of action and the events triggered by curcumin are different from those occurring during typical apoptosis.


Assuntos
Apoptose/fisiologia , Membrana Celular/fisiologia , Membrana Celular/ultraestrutura , Curcumina/farmacologia , Membranas Intracelulares/fisiologia , Mitocôndrias/fisiologia , Timo/fisiologia , Animais , Anexina A6 , Transporte Biológico/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Células Cultivadas , Curcumina/farmacocinética , DNA/metabolismo , Dexametasona/farmacologia , Fluoresceína-5-Isotiocianato , Membranas Intracelulares/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Fosfatidilserinas/farmacologia , Propídio/farmacocinética , Ratos , Ratos Endogâmicos WKY , Timo/efeitos dos fármacos , Timo/ultraestrutura
11.
Pol Arch Med Wewn ; 93(2): 122-9, 1995 Feb.
Artigo em Polonês | MEDLINE | ID: mdl-7479229

RESUMO

UNLABELLED: The aim of the study was to determine what is the correlation between immunological phenomena and pathological changes in chronic gastritis and how it refers to the histology and endoscopy. PATIENTS AND METHODS: 42 patients with dyspepsia underwent following procedure: 1) gastroscopy and antral biopsy of 3 specimens; 2) histology; 3) immunofluorescence of the specimens in purpose to detect bound immunoglobulins using antibodies anti-human IgA + IgG + IgM labelled with rhodamine; 4) serologic test for anti-H.pylori antibodies. The research included 17 females and 25 males (ages 30-86, median 53.4 +/- 15.47). The obtained data were compared referring to mutual correlations and presented according to the kind of pathological changes depending on:--presence or absence of Helicobacter pylori infection;--presence or absence of anti-Helicobacter pylori antibodies;--presence or absence of lymphocytic infiltration;--presence or absence of bound immunoglobulins in gastric mucosa. RESULTS: --We have not observed ulcerations in anti-Helicobacter pylori seronegative group;--intestinal metaplasia and gastric ulcer were more frequent in bound immunoglobulins positive group;--biliary reflux was observed less frequently in lymphocyte infiltration negative group then in the positive one. CONCLUSION: pathological changes in chronic gastritis may depend not only on the presence of Helicobacter pylori infection, but also on the presence and quality of immunological response on its antigens.


Assuntos
Mucosa Gástrica/patologia , Gastrite/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antibacterianos/imunologia , Biópsia , Doença Crônica , Feminino , Imunofluorescência , Gastrite/microbiologia , Gastrite/patologia , Gastroscopia , Infecções por Helicobacter/patologia , Humanos , Masculino , Pessoa de Meia-Idade
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