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1.
J. appl. oral sci ; 23(5): 536-546, Sept.-Oct. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-764159

RESUMO

In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity. Recently, our research group has reported a differential immunogenicity when T lymphocytes were stimulated with these different serotypes. In particular, it was demonstrated that the serotype b of A. actinomycetemcomitans has a stronger capacity to trigger Th1- and Th17-type cytokine production.Objective This study aimed to quantify the expression of different CC chemokines (CCLs) and receptors (CCRs) in T lymphocytes stimulated with the differentA. actinomycetemcomitans serotypes. In addition, the expression of the transcription factors T-bet, GATA-3, RORC2, and Foxp3, master-switch genes implied in the Th1, Th2, Th17, and T-regulatory differentiation, respectively, was analysed in order to determine T-cell phenotype-specific patterns of CCL and CCR expression upon A. actinomycetemcomitans stimulation.Material and Methods Human naïve CD4+ T lymphocytes were obtained from healthy subjects and stimulated with autologous dendritic cells primed with the differentA. actinomycetemcomitans serotypes. The expression levels for the chemokines CCL1, CCL2, CCL3, CCL5, CCL11, CCL17, CCL20, CCL21, CCL25, and CCL28, as well as the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 were quantified by qPCR. Similarly, the expression levels for the transcription factors T-bet, GATA-3, RORC2, and Foxp3 were quantified and correlated with the CCL and CCR expression levels.Results Higher expression levels of CCL2, CCL3, CCL5, CCL20, CCL21, CCL28, CCR1, CCR2, CCR5, CCR6, CCR7, and CCR9 were detected in T lymphocytes stimulated with the serotype b of A. actinomycetemcomitans compared with the other serotypes. In addition, these higher expression levels of CCLs and CCRs positively correlated with the increased levels of T-bet and RORC2 when T lymphocytes were stimulated with the serotype b.Conclusion A T-lymphocyte response biased towards a Th1- and Th17-pattern of CCL and CCR expression was detected under stimulation with the serotype b ofA. actinomycetemcomitans.


Assuntos
Adulto , Feminino , Humanos , Masculino , Adulto Jovem , Aggregatibacter actinomycetemcomitans/imunologia , Antígenos de Diferenciação de Linfócitos T/análise , Quimiocinas CC/análise , Receptores CCR/análise , Linfócitos T/imunologia , Aggregatibacter actinomycetemcomitans/genética , Antígenos de Diferenciação de Linfócitos T/genética , Antígenos de Diferenciação de Linfócitos T/imunologia , Células Cultivadas , Diferenciação Celular/imunologia , Quimiocinas CC/genética , Quimiocinas CC/imunologia , Células Dendríticas/imunologia , Citometria de Fluxo , Ativação Linfocitária , Reação em Cadeia da Polimerase , Receptores CCR/genética , Receptores CCR/imunologia , Sorogrupo
2.
Braz. j. infect. dis ; 19(4): 403-409, July-Aug. 2015. tab, ilus
Artigo em Inglês | LILACS | ID: lil-759282

RESUMO

Human cytomegalovirus is a ubiquitous pathogen that infects the majority of the world's population. After long period of time co-evolving with human being, this pathogen has developed several strategies to evade host immune surveillance. One of the major trick is encoding homologous to those of the host organism or stealing host cellular genes that have significant functions in immune system. To date, we have found several viral immune analogous which include G protein coupled receptor, class I major histocompatibility complex and chemokine. Chemokine is a small group of molecules which is defined by the presence of four cysteines in highly conserved region. The four kinds of chemokines (C, CC, CXC, and CX3C) are classified based on the arrangement of 1 or 2 N-terminal cysteine residues. UL128 protein is one of the analogous that encoded by human cytomegalovirus that has similar amino acid sequences to the human CC chemokine. It has been proved to be one of the essential particles that involved in human cytomegalovirus entry into epithelial/endothelial cells as well as macrophages. It is also the target of potent neutralizing antibodies in human cytomegalovirus-seropositive individuals. We had demonstrated the chemotactic trait of UL128 protein in our previous study. Recombinant UL128 in vitrohas the ability to attract monocytes to the infection region and enhances peripheral blood mononuclear cell proliferation by activating the MAPK/ERK signaling pathway. However, the way that this viral encoded chemokine interacting with peripheral blood mononuclear cells and the detailed mechanism that involving the virus entry into host cells keeps unknown. Here we performed in vitroinvestigation into the effects of UL128 protein on peripheral blood mononuclear cell's activation and receptor binding, which may help us further understand the immunomodulatory function of UL128 protein as well as human cytomegalovirus diffusion mechanism.


Assuntos
Humanos , Quimiocinas CC , Citomegalovirus , Regulação Viral da Expressão Gênica/genética , Leucócitos Mononucleares/virologia , Glicoproteínas de Membrana/imunologia , Proteínas do Envelope Viral/imunologia , Células Cultivadas , Reagentes para Ligações Cruzadas , Quimiocinas CC/genética , Quimiocinas CC/imunologia , Citomegalovirus/genética , Citomegalovirus/imunologia , Receptores de Quimiocinas/genética , Proteínas Recombinantes/imunologia
3.
Braz. j. otorhinolaryngol. (Impr.) ; 77(5): 605-610, Sept.-Oct. 2011. ilus
Artigo em Inglês | LILACS | ID: lil-601859

RESUMO

Glucocorticoids are considered the main treatment option for nasal polyps, but their effect is only recently being understood. AIM: To evaluate whether fluticasone propionate (FP) inhibits the inflammatory process induced by TNF-alpha in vitro, and to assess if NF-kappaB is associated to this inhibition. STUDY DESIGN: Experimental in vitro study. MATERIALS AND METHODS: Nasal polyp fibroblasts were cultured during 24 hours. Three different concentrations of FP (1, 10 and 100 nM, added to TNF-alpha) were compared to negative (without additive) and positive (TNF-alpha) controls. Gene expression (RTQ-PCR) and protein concentration (ELISA) of VCAM-1, ICAM-1, eotaxin and RANTES were measured, as well as the nuclear translocation of NF-kappaB. RESULTS: TNF-alpha significantly increased protein concentration and RNA expression of all the studied molecules, as well as the nuclear translocation of NF-kappaB, when compared to the negative control. FP decreased these parameters in a dose-dependent manner, statistically different from positive control up to 100nM. CONCLUSIONS: FP extensively inhibited inflammatory recruiters, at both protein and RNA levels, confirming the ability of glucocorticoids to modulate the inflammatory process in nasal polyps. This inhibition was associated to decreased NF-kappaB nuclear translocation, demonstrating that this is an important mechanism of glucocorticoids action for nasal polyps.


Glicocorticoides são considerados a principal opção terapêutica para polipose nasossinusal, mas seus efeitos estão sendo descobertos apenas recentemente. OBJETIVO: Avaliar se proprionato de fluticasona (FP) inibe in vitro o processo inflamatório induzido por TNF-alfa, e se NF-kappaB está associado a esta inibição. FORMA DE ESTUDO: Experimental in vitro. MATERIAIS E MÉTODOS: Fibroblastos de pólipos nasais foram cultivados por 24 horas. Três concentrações diferentes de FP (1, 10 e 100nM, além do TNF-alfa) foram comparados a controles negativo (sem aditivo) e positivo (TNF-alfa). Expressão gênica (RTQ-PCR) concentração proteica (ELISA) de VCAM-1, ICAM-1, eotaxin e RANTES foram medidos, assim como a translocação nuclear de NF-kappaB. RESULTADOS: TNF-alfa aumentou significativamente a concentração proteica e expressão gênica de todas molé¬culas estudadas, assim como a translocação nuclear de NF-kappaB, quando comparado ao controle negativo. O FP diminuiu estes parâmetros numa forma dose-dependente, diferente estatisticamente do controle positivo até 100nM. CONCLUSÕES: O FP extensivamente inibiu os recrutadores inflamatórios, em níveis proteicos e gênicos, confirmando a habilidade dos glicocorticoides em modular o processo inflamatório na polipose nasossinusal. Esta inibição esteve associada à diminuição da translocação nuclear de NF-kappaB, demonstrando que este é um importante mecanismo de ação dos glicocorticoide na polipose nasossinusal.


Assuntos
Humanos , Androstadienos/farmacologia , Anti-Inflamatórios/farmacologia , Fibroblastos/efeitos dos fármacos , Pólipos Nasais/tratamento farmacológico , Células Cultivadas , Moléculas de Adesão Celular/metabolismo , Quimiocinas CC/metabolismo , Fibroblastos/patologia , NF-kappa B/metabolismo , Pólipos Nasais/patologia , Fator de Necrose Tumoral alfa/metabolismo
4.
Braz. dent. j ; 19(1): 3-8, 2008. tab
Artigo em Inglês | LILACS | ID: lil-481120

RESUMO

Advances in diagnostic research are moving towards methods whereby the periodontal risk can be identified and quantified by objective measures using biomarkers. Patients with periodontitis may have elevated circulating levels of specific inflammatory markers that can be correlated to the severity of the disease. The purpose of this study was to evaluate whether differences in the serum levels of inflammatory biomarkers are differentially expressed in healthy and periodontitis patients. Twenty-five patients (8 healthy patients and 17 chronic periodontitis patients) were enrolled in the study. A 15 mL blood sample was used for identification of the inflammatory markers, with a human inflammatory flow cytometry multiplex assay. Among 24 assessed cytokines, only 3 (RANTES, MIG and Eotaxin) were statistically different between groups (p<0.05). In conclusion, some of the selected markers of inflammation are differentially expressed in healthy and periodontitis patients. Cytokine profile analysis may be further explored to distinguish the periodontitis patients from the ones free of disease and also to be used as a measure of risk. The present data, however, are limited and larger sample size studies are required to validate the findings of the specific biomarkers.


Avanços no diagnóstico da doença periodontal levam a métodos nos quais o risco e atividade da doença periodontal podem ser identificados e quantificados por biomarcadores. Pacientes com periodontite podem apresentar elevados níveis circulatórios de marcadores inflamatórios específicos que podem ser correlacionados com a severidade da doença. Portanto, o objetivo desse estudo foi avaliar as diferenças nos níveis séricos de biomarcadores inflamatórios em pacientes saudáveis e com doença periodontal. Foram incluídos no estudo 25 pacientes (8 saudáveis e 17 com periodontite crônica). Uma amostra de 15 mL de sangue foi obtida para identificar os marcadores inflamatórios simultaneamente utilizando Array de proteínas através de citometria de fluxo. De 24 citocinas inflamatórias analisadas, apenas 3 (RANTES, MIG e Eotaxina) apresentaram diferenças estatisticamente significantes (p<0,05) entre os dois grupos. Conclui-se que alguns marcadores inflamatórios selecionados apresentam diferença de concentração em pacientes com periodontite e saudáveis. A análise do perfil de citocinas pode ser utilizada tanto para distinguir pacientes periodontais de pacientes saudáveis, como para medir o risco à doença. Contudo, mais estudos com número maior de amostras são necessários para validar os achados sobre os biomarcadores específicos.


Assuntos
Humanos , Periodontite Crônica/sangue , Mediadores da Inflamação/sangue , Biomarcadores/sangue , /sangue , /sangue , /sangue , /sangue , Quimiocina CXCL9/sangue , Quimiocinas CC/sangue , Citocinas/sangue , Proteína Ligante Fas/sangue , /sangue , Hemorragia Gengival/sangue , Fator Estimulador de Colônias de Granulócitos/sangue , Fator Estimulador de Colônias de Granulócitos e Macrófagos/sangue , Interferon gama/sangue , Interleucina-9/sangue , Interleucinas/sangue , Linfotoxina-alfa/sangue , Perda da Inserção Periodontal/sangue , Bolsa Periodontal/sangue , Fator de Crescimento Transformador beta/sangue
5.
Rio de Janeiro; s.n; 29 nov. 2006. xiii,134 p. ilus, graf.
Tese em Português | LILACS | ID: lil-449198

RESUMO

Os linfócitos T gd apresentam um papel importante durante reações alérgicas, desempenhando funções efetoras e regulatórias.Quimiocinas e moléculas de adesão são fundamentais para a migração de linfócitos em resposta a diversos estímulos.No entanto, os mecanismos pelos quais os linfócitos T migram para o sítio inflamatório durante reações de origem alérgica não foram descritos.Neste estudo, pretendemos investigar o papel das quimiocinas, selectinas e integrinas na migração de linfócitos T após o desafio intratorácico com ovoalbumina em camundongos previamente sensibilizados.O estímulo com ovoalbumina induziu um aumento no número de linfócitos T na cavidade pleural após 12 horas, permanecendo significativo até 96 horas, acompanhado por um aumento no sangue em 48 horas. O desafio antigênico também induziu um aumento no número de linfócitos T expressando CCR2, CCR5 e CCR9 na pleura entre 12 e 72 horas e no sangue em 48 horas, assim como o aumento da expressão dessas moléculas no linfonodo torácico 48 horas após o estímulo.Os níveis de cc quimiocinas CCL2, CCL3, CCL5 e CCL25 encontraram-se aumentados nos lavados pleurais recolhidos 24 horas após o estímulo, e foram produzidas principalmente por macrófagos pleurais...A neutralização in vitro de CCL2 inibiu a quimiotaxia de linfócitos Td induzida por lavados pleurais de animais estimulados, confirmando o papel direto desta quimiocina nos linfócitos Td.Confirmando este dado, o desafio antigênico falhou em induzir um aumento no número de linfócitos T d na cavidade pleural, no sangue e em linfonodos de camundongos deficientes de CCR2.Foi também observado um aumento na expressão de L-selectina em linfócitos T ?d recolhidos dos linfonodos torácicos de animais desafiados. Linfócitos T recolhidos do sangue de animais estimulados apresentaram uma expressão maior de L-selectina quando comparados com as células da cavidade pleural, sugerindo o shedding desta molécula.O desafio antigênico também induziu um acúmulo de linfócito...


Assuntos
Animais , Camundongos , Quimiocinas , Quimiocinas CC , Linfócitos , Linfócitos T , Camundongos
6.
Braz. j. med. biol. res ; 39(4): 441-445, Apr. 2006. ilus
Artigo em Inglês | LILACS | ID: lil-425080

RESUMO

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the human central nervous system. Although its etiology is unknown, the accumulation and activation of mononuclear cells in the central nervous system are crucial to its pathogenesis. Chemokines have been proposed to play a major role in the recruitment and activation of leukocytes in inflammatory sites. They are divided into subfamilies on the basis of the location of conserved cysteine residues. We determined the levels of some CC and CXC chemokines in the cerebrospinal fluid (CSF) of 23 relapsing-remitting MS patients under interferon-ß-1a therapy and 16 control subjects using ELISA. MS patients were categorized as having active or stable disease. CXCL10 was significantly increased in the CSF of active MS patients (mean ± SEM, 369.5 ± 69.3 pg/mL) when compared with controls (178.5 ± 29.1 pg/mL, P < 0.05). CSF levels of CCL2 were significantly lower in active MS (144.7 ± 14.4 pg/mL) than in controls (237.1 ± 16.4 pg/mL, P < 0.01). There was no difference in the concentration of CCL2 and CXCL10 between patients with stable MS and controls. CCL5 was not detectable in the CSF of most patients or controls. The qualitative and quantitative differences of chemokines in CSF during relapses of MS suggest that they may be useful as a marker of disease activity and of the mechanisms involved in the pathogenesis of the disease.


Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Quimiocinas CC/líquido cefalorraquidiano , Quimiocinas CXC/líquido cefalorraquidiano , Esclerose Múltipla Recidivante-Remitente/líquido cefalorraquidiano , Adjuvantes Imunológicos/uso terapêutico , Biomarcadores/líquido cefalorraquidiano , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Interferon beta/uso terapêutico , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico
7.
Gac. méd. Méx ; 142(2): 139-144, mar.-abr. 2006. ilus
Artigo em Espanhol | LILACS | ID: lil-570743

RESUMO

Durante la última década se han descubierto tres péptidos con actividad quimotáctica específica para los eosinófilos y que son miembros de la familia de las quimocinas. Estas citocinas inducen a los eosinófilos a realizar diferentes funciones como quimotaxis, migración transendotelial e inducción de la liberación de radicales de oxígeno. Como los eosinófilos infiltran tanto las vías aéreas de pacientes asmáticos como los pólipos nasales, se ha postulado que las eotaxinas pueden ser responsables del reclutamiento de estas células. Los eosinófilos tienen la propiedad de inducir remodelamiento de la matriz extracelular y daño tisular a través de la liberación de proteasas tóxicas, mediadores inflamatorios, citocinas y radicales de oxígeno. Por lo cual, el desarrollo de estrategias terapéuticas que inhiban el reclutamiento de estas células constituye una esperanza en el tratamiento de las enfermedades alérgicas. Este artículo revisa la función de las eotaxinas en asma y poliposis nasal, además de discutir el posible uso de antagonistas de CCR3, receptor de las eotaxinas, como una nueva modalidad terapéutica de asma y poliposis nasal.


Over the last few years, three specific eosinophil activating peptides, eotaxin-1, -2 and -3, members of the chemokine family have been identified. These cytokines exert a number of functions on eosinophils including chemotaxis, transendothelial migration and induction of the release of reactive oxygen species. Eosinophils are considered to play an important role in allergic disease by causing tissue damage through the release of toxic proteases, lipid mediators, cytokines and oxygen free radicals. This article reviews the role of eotaxins in asthma and nasal polyps. Discussion focuses on therapeutic guidelines, particularly as it has been shown that CCR3, the major chemokine receptor in eosinophils, serves as a eotaxin receptor.


Assuntos
Humanos , Asma/etiologia , Pólipos Nasais/etiologia , Quimiocinas CC/fisiologia
8.
Mem. Inst. Oswaldo Cruz ; 100(supl.1): 93-96, Mar. 2005.
Artigo em Inglês | LILACS | ID: lil-402181

RESUMO

The comprehension of the pathogenesis of Trypanosoma cruzi-elicited myocarditis is crucial to delineate new therapeutic strategies aiming to ameliorate the inflammation that leads to heart dysfunction, without hampering parasite control. The augmented expression of CCL5/RANTES and CCL3/MIP-1alpha, and their receptor CCR5, in the heart of T. cruzi-infected mice suggests a role for CC-chemokines and their receptors in the pathogenesis of T. cruzi-elicited myocarditis. Herein, we discuss our recent results using a CC-chemokine receptor inhibitor (Met-RANTES), showing the participation of CC-chemokines in T. cruzi infection and unraveling CC-chemokine receptors as an attractive therapeutic target for further evaluation in Chagas disease.


Assuntos
Animais , Camundongos , Cardiomiopatia Chagásica/tratamento farmacológico , /análogos & derivados , Quimiocinas CC/metabolismo , Miocardite/tratamento farmacológico , Receptores de Quimiocinas/antagonistas & inibidores , Trypanosoma cruzi , /imunologia , /imunologia , Cardiomiopatia Chagásica/imunologia , Cardiomiopatia Chagásica/metabolismo , /uso terapêutico , Quimiotaxia de Leucócito/imunologia , Miocardite/imunologia , Miocardite/metabolismo , Miocardite/parasitologia , Trypanosoma cruzi/imunologia
9.
P. R. health sci. j ; 19(4): 345-351, Dec. 2000.
Artigo em Inglês | LILACS | ID: lil-334082

RESUMO

The duration from initial infection with HIV-1 to CD4 lymphocyte depletion and progression to AIDS varies among infected individuals. Despite treatment with highly active antiretroviral therapy (HAART), patients still show different stages of disease progression. We examined the role of beta-chemokines and its receptor, CCR5 in HIV-1 infected children in order to define determinants of HIV progression among treated individuals. Population was divided in two groups: Group 1--Long Term Non Progressors (LTNP) includes 10 patients with B1-B2 CDC disease classification and with a less aggressive therapy (only 2 in HAART); Group 2--Rapid Progressors (RP) includes 9 patients with C3 disease classification. All the patients had a CCR5 wild type (wt) genotype indicating that they do not have the 32 base-pair deletion associated with slower progression. There was an increased production of MIP 1-beta in 8/10 LTNP but only in 4/9 Progressors (Paired t-test/Wilcoxon Sign test, p-value < 0.05). The change in the levels of MIP-1 beta after PHA stimulation was statistically significant in both groups. The levels of RANTES increased in LTNP and RP and the change of the levels after mitogen stimulation was statistically significant for both groups included. The production of RANTES and MIP-1 beta in response to stimulation between both groups was not statistically significant. The production of MIP-1 alpha was variable in both groups and the difference in the levels after mitogen stimulation between the groups was not statistically significant. These results suggest that beta-chemokines do not play an important role in HIV-1 progression in children undergoing HAART.


Assuntos
Criança , Humanos , Quimiocinas CC , Infecções por HIV/imunologia , Receptores CCR5
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