RESUMO
The immune mechanisms that contribute to the efficacy of treatment of cutaneous leishmaniasis (CL) are not fully understood. The aim of this study was to define immune correlates of the outcome of treatment of CL caused by Leishmania (Viannia) species during standard of care treatment with pentavalent antimonials. We conducted a comparative expression profiling of immune response genes in peripheral blood mononuclear cells (PBMCs) and lesion biopsy specimens obtained from CL patients before and at the end of treatment (EoT) with meglumine antimoniate. The ex vivo response of PBMCs to L (V) panamensis partially reflected that of lesion microenvironments. Significant downregulation of gene expression profiles consistent with local innate immune responses (monocyte and neutrophil activation and chemoattractant molecules) was observed at EoT in biopsy specimens of patients who cured (n = 8), compared to those from patients with treatment failure (n = 8). Among differentially expressed genes, pretreatment expression of CCL2 was significantly predictive of the therapeutic response (receiver operating characteristic [ROC] curve, area under the curve [AUC] = 0.82, P = 0.02). Polymorphisms in regulatory regions of the CCL2 promoter were analyzed in a pilot cohort of DNA samples from CL patients (cures, n = 20, and treatment failure, n = 20), showing putative association of polymorphisms rs13900(C/T) and rs2857656(G/C) with treatment outcome. Our data indicate that dampening gene expression profiles of monocyte and neutrophil activation characterize clinical cure after treatment of CL, supporting participation of parasite-sustained inflammation or deregulated innate immune responses in treatment failure.
Assuntos
Antiprotozoários/uso terapêutico , Citocinas/metabolismo , Imunidade Inata/fisiologia , Leishmania/imunologia , Leishmaniose/tratamento farmacológico , Leishmaniose/imunologia , Antimoniato de Meglumina/uso terapêutico , Perfilação da Expressão Gênica , Humanos , Leishmaniose/metabolismo , Leucócitos Mononucleares/metabolismo , Proteínas Quimioatraentes de Monócitos/metabolismo , Monócitos/metabolismoRESUMO
Introduction: Reduction in the expression of inflammatory markers and oxidative stress associated with exercise will protect against cardiovascular complications in Diabetes Mellitus (DM). Aim: The aim of this study was evaluated cardiovascular fitness (VO2 Max), interleukin-6 (IL-6), monocyte chemo-attractant protein 1 (MCP-1) and serum lipid peroxidation (TBARS) in overweight patients with Type-1 diabetes (T1DM) participating in a lifestyle-change program. Results: 20 T1DM overweight patients (43.3 ± 13.8 years), with BMI= 29.6 ± 3.5 kg/m2 , initial HbA1c 7.9 ± 0.91% and treated with multiple insulin injections, were included in this work. The lifestyle-change program consisted of: a) walking 10,000 steps/day, b) sequence of exercises of 24 minutes, 3-5 times/week, c) ¨healthy-plate¨ (and counting carbohydrates, and d) prandial insulin as blood-glucose levels. VO2 max, HbA1c, TBARS, IL6, MCP-1 were determined before starting the lifestyle-change program. Six months of adherence later, participants showed an average number of steps of 8242 ± 1834, a significant increase in VO2 max, (33.4 ±1.3 vs 36.2 ±1.5 ml.Kg-1.min-1 p= 0.008), a significant decrease in serum MCP-1 (314 ±42 vs 235 ±43 MFI p= 0.02), and less TBARS (3.01 ±0.44 vs 2.12 ±0.22 µmol/mL p= 0.015). IL-6 and HbA1c showed no significant decrease. Conclusion: Our results showed that a 6-month systemized and simple exercise plan improves cardiorespiratory fitness (VO2 max), and reduces both circulating oxidative stress and inflammation markers in overweight patients with T1DM.
Introducción: La reducción en la expresión de marcadores inflamatorios y de estrés oxidativo asociado con el ejercicio podría proteger contra las complicaciones cardiovasculares de la diabetes mellitus (DM). Objetivo: El objetivo de este estudio fue evaluar en pacientes con DM tipo1 (DMT1) y sobrepeso, la capacidad cardiorespiratoria (VO2 Max), la expresión sérica de marcadores inflamatorios (IL-6 y MCP-1) y la peroxidación lipídica sérica (TBARS), luego de participar por 6 meses de un programa de cambios de estilo de vida. Resultados: Veinte pacientes adultos (43.3 ± 13.8 años), de ambos sexos, con un Índice de Masa Corporal de 29.6 ± 3.5 kg/m2 , HbA1c inicial de 7,9% ± 0,91, en tratamiento con inyecciones múltiples de insulina participaron del estudio. Se indicó: 1) caminar 10.000 pasos/día, 2) realizar en domicilio una secuencia de ejercicios de 20 minutos, 3-5 veces/semana, 3) plato saludable (consumo de 1 fruta antes de las 3 comidas principales), 4) Insulina prandial según glucemia y conteo de carbohidratos. Se registraron parámetros antropométricos, presión arterial, se determinó VO2 max, y se midieron los niveles séricos de HbA1c, IL6, MCP-1 y TBARs. Luego de seis meses, los participantes alcanzaron un número promedio de pasos de 8242 ± 1834 y mostraron un aumento significativo en VO2 max, (33.4 ±1.3 vs 36.2 ±1.5 ml.Kg-1.min-1 p= 0.008). Además, se encontró una disminución significativa de MCP-1 (314 ±42 vs 235 ±43 MFI p=0.02) y TBARs (3.01 ±0.44 vs 2.12 ±0.22 µmol/mL p= 0.015) en comparación con el día 0. No se observaron modificaciones en los niveles de IL-6 y HbA1c. Conclusión: Nuestros resultados demuestran que el ejercicio, implementado como un plan accesible y acompañado, es adecuado para reducir los riesgos de inflamación y estado pro-oxidativo en pacientes con DM tipo1.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Exercício Físico/fisiologia , Diabetes Mellitus Tipo 1/terapia , Sobrepeso/terapia , Consumo de Oxigênio/fisiologia , Biomarcadores , Peroxidação de Lipídeos , Interleucina-6/sangue , Estresse Oxidativo , Proteínas Quimioatraentes de Monócitos , Diabetes Mellitus Tipo 1/fisiopatologia , Sobrepeso/fisiopatologia , Aptidão Cardiorrespiratória/fisiologia , Inflamação , Estilo de VidaRESUMO
OBJECTIVES: The aim of this study was to investigate the association between dairy product consumption and plasma inflammatory biomarkers levels among a representative sample of Brazilian adults from São Paulo City. METHODS: Data were acquired from the Health Survey for São Paulo, a cross-sectional population-based study. All individuals 20 to 59 y of age with complete food consumption information (24-h dietary recall and food frequency questionnaire) and blood sample analysis were included (N = 259). The sample was separated into two groups according to systemic inflammatory pattern considering plasma levels of C-reactive protein; tumor necrosis factor-α; soluble intracellular adhesion molecule; soluble vascular cell adhesion molecule, monocyte chemoattractant protein; interleukin-1ß, -6, -8, -10, and -12; adiponectin; leptin; and homocysteine. Multiple logistic regression tests were conducted to estimate the odds ratio for the inflammatory cluster across tertiles of dairy consumption. RESULTS: When adjusted by age, smoking status, and energy intake the odds ratio for the inflammatory cluster group in the highest tertile of yogurt consumption was 0.34 (95% confidence interval [CI], 0.14-0.81) relative to the reference tertile, demonstrating also a linear effect (Ptrend = 0.015). Cheese consumption exhibited an odds ratio of 2.49 (95% CI, 1.09-5.75) relative to the reference. CONCLUSIONS: Increasing yogurt consumption might have a protective effect on inflammation, whereas cheese consumption appears to be associated with a proinflammatory status. The results of the present study aggregate a new perspective on existing evidence demonstrating the importance of assessing the contribution of dairy products on diet and their effect on the development of non-communicable diseases and associated risk factors.
Assuntos
Queijo/efeitos adversos , Laticínios/efeitos adversos , Dieta/efeitos adversos , Mediadores da Inflamação/sangue , Iogurte/efeitos adversos , Adiponectina/sangue , Adulto , Brasil/epidemiologia , Proteína C-Reativa/análise , Moléculas de Adesão Celular/sangue , Estudos Transversais , Inquéritos sobre Dietas , Ingestão de Energia , Feminino , Inquéritos Epidemiológicos , Homocisteína/sangue , Humanos , Interleucinas/sangue , Leptina/sangue , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Proteínas Quimioatraentes de Monócitos/sangue , Doenças não Transmissíveis/epidemiologia , Razão de Chances , Fatores de Risco , Fator de Necrose Tumoral alfa/sangue , Adulto JovemRESUMO
Galectin-8 (Gal-8) is a mammalian ß-galactoside-binding lectin, endowed with proinflammatory properties. Given its capacity to enhance antigen-specific immune responses in vivo, we investigated whether Gal-8 was also able to promote APC activation to sustain T cell activation after priming. Both endogenous [dendritic cells (DCs)] and bone marrow-derived DCs (BMDCs) treated with exogenous Gal-8 exhibited a mature phenotype characterized by increased MHC class II (MHCII), CD80, and CD86 surface expression. Moreover, Gal-8-treated BMDCs (Gal-8-BMDCs) stimulated antigen-specific T cells more efficiently than immature BMDCs (iBMDCs). Proinflammatory cytokines IL-3, IL-2, IL-6, TNF, MCP-1, and MCP-5, as well as growth factor G-CSF, were augmented in Gal-8-BMDC conditioned media, with IL-6 as the most prominent. Remarkably, BMDCs from Gal-8-deficient mice (Lgals8-/- BMDC) displayed reduced CD86 and IL-6 expression and an impaired ability to promote antigen-specific CD4 T cell activation. To test if Gal-8-induced activation correlates with the elicitation of an effective immune response, soluble Gal-8 was coadministrated with antigen during immunization of BALB/cJ mice in the experimental foot-and-mouth disease virus (FMDV) model. When a single dose of Gal-8 was added to the antigen formulation, an increased specific and neutralizing humoral response was developed, sufficient to enhance animal protection upon viral challenge. IL-6 and IFN-γ, as well as lymphoproliferative responses, were also incremented in Gal-8/antigen-immunized animals only at 48 h after immunization, suggesting that Gal-8 induces the elicitation of an inflammatory response at an early stage. Taking together, these findings argue in favor of the use of Gal-8 as an immune-stimulator molecule to enhance the adaptive immune response.
Assuntos
Apresentação de Antígeno , Antígenos Virais/imunologia , Linfócitos T CD4-Positivos/imunologia , Células Dendríticas/imunologia , Febre Aftosa/imunologia , Galectinas/imunologia , Imunidade Adaptativa , Animais , Antígenos Virais/administração & dosagem , Antígenos Virais/genética , Linfócitos T CD4-Positivos/virologia , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Células Dendríticas/virologia , Febre Aftosa/genética , Febre Aftosa/prevenção & controle , Febre Aftosa/virologia , Vírus da Febre Aftosa/crescimento & desenvolvimento , Vírus da Febre Aftosa/imunologia , Galectinas/genética , Galectinas/farmacologia , Regulação da Expressão Gênica , Fator Estimulador de Colônias de Granulócitos/genética , Fator Estimulador de Colônias de Granulócitos/imunologia , Imunização , Interleucina-2/genética , Interleucina-2/imunologia , Interleucina-3/genética , Interleucina-3/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Quimioatraentes de Monócitos/genética , Proteínas Quimioatraentes de Monócitos/imunologia , Transdução de Sinais , Fatores de Tempo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Chemokines are important mediators of immunological responses during inflammation and under steady-state conditions. In addition to regulating cell migration, some chemotactic cytokines have direct effects on bacteria. Here, we characterized the antibacterial ability of the synthetic oligopeptide CCL1357-75, which corresponds to the carboxyl-terminal region of the human chemokine CCL13. In vitro measurements indicated that CCL1357-75 disrupts the cell membrane of Pseudomonas aeruginosa through a mechanism coupled to an unordered-helicoidal conformational transition. In a murine pneumonic model, CCL1357-75 improved mouse survival and bacterial clearance and decreased neutrophil recruitment, proinflammatory cytokines and lung pathology compared with that observed in untreated infected animals. Overall, our study supports the ability of chemokines and/or chemokine-derived oligopeptides to act as direct defense agents against pathogenic bacteria and suggests their potential use as alternative antibiotics.
Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Pneumonia Bacteriana/tratamento farmacológico , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/efeitos dos fármacos , Animais , Antibacterianos/uso terapêutico , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Modelos Animais de Doenças , Humanos , Camundongos , Proteínas Quimioatraentes de Monócitos/química , Proteínas Quimioatraentes de Monócitos/metabolismo , Proteínas Quimioatraentes de Monócitos/farmacologia , Oligopeptídeos/farmacologia , Oligopeptídeos/uso terapêutico , Fragmentos de Peptídeos , Domínios ProteicosRESUMO
O objetivo do presente estudo foi avaliar os níveis das metaloproteinases da matriz MMP-9, MMP-3 e MMP-13, e das quimiocinas MCP-1, MIP-1β e RANTES, no fluido gengival de dentes sob força ortodôntica. Foram recrutados 14 pacientes (3 homens e 11 mulheres) que foram submetidos à movimentação ortodôntica dos caninos superiores. Amostras do fluido gengival foram coletadas com tiras de Periopaper em diferentes tempos. O volume do FG foi determinado com o uso do Periotron® e os niveis das MMPs e das quimiocinas quantificados usando-se uma multianálise imunoenzimática com microesferas. Os resultados mostraram que existe um aumento significativo no volume do FG na área de pressão em quase todos os tempos analisados, quando comparados às áreas de tensão. Os níveis da MMP-9 foram muito superiores aos das MMP-13 e MMP-3. Na análise da evolução do tempo pode ser observada uma alteração significativa nos níveis da quantidade total de MMP-9, MMP-13 e MMP-3 e na concentração de MMP-13 e MMP-3 durante o período de movimentação dentária no lado de pressão. A elevação dos níveis de expressão destas MMPs 1 hora após a aplicação da força ortodôntica sugere que estas enzimas estejam envolvidas na remodelação periodontal induzida. As quimiocinas MCP-1, MIP-1β e RANTES foram detectadas no sulco gengival em todos os diferentes intervalos de tempo analisados, nas áreas de tensão e de pressão. Porém, diversas amostras estavam abaixo do nível de detecção do ensaio e, os níveis dessas quimiocinas no fluido gengival não parecem ser alterados pelas forças ortodônticas.
The goal of the present study was to evaluate the levels of the matrix metalloproteinases MMP-9, MMP-3 and MMP-13 and of the chemokines MCP-1, MIP-1β and RANTES in the gingival crevicular fluid of teeth under orthodontic forces. Fourteen subjects (3 males and 11 females) were enrolled and subjected to orthodontic tooth movement of their maxillary canines. Samples of gingival crevicular fluid were collected from both tension and pressure sides using Periopaper strips at different time points. The volume of GCF was determined using a Periotron® and the levels of MMPs and chemokines quantified using a multiplex microbead immunoassay. The results demonstrated that the levels of MMP-9 were higher than the levels of MMP-13 and MMP-3. Statistically significant fluctuations during the orthodontic tooth movement could be detected for the total amount of MMP-9, MMP-13 and MMP-3 and for the concentration of MMP-13 and MMP-3 at the pressure side. The elevation in the levels of these MMPs, 1 hour after the application of the orthodontic force, suggests that these enzymes are involved in the induced periodontal remodeling. The chemokines MCP-1, MIP-1β and RANTES were detected in the GCF in both tension and pressure sides at different time points. However, several samples were below the level of detection of the assay and the levels of theses mediators in GCF did not seem to be altered by the orthodontic forces.
Assuntos
Humanos , Citocinas , Proteínas Inflamatórias de Macrófagos , Metaloproteinase 9 da Matriz , Proteínas Quimioatraentes de Monócitos , Técnicas de Movimentação Dentária/efeitos adversos , Ortodontia Corretiva/métodos , Dente Canino , Líquido do Sulco Gengival/química , Monitoramento Ambiental , Interpretação Estatística de DadosRESUMO
Small volumes of cervical secretions have limited measurements of immunity at the cervix, which may be important to studies of human papillomavirus (HPV). We report the use of recycling immunoaffinity chromatography to efficiently study immune profiles in cervical secretions. Frozen pairs of plasma and cervical secretions (collected on ophthalmic sponges) were selected randomly from women with normal cervical cytology (n = 50) participating in a natural history study of HPV in Guanacaste, Costa Rica. Single 25- micro l aliquots of plasma and (diluted) cervical secretions were assayed for interleukin (IL) -1 beta, -2, -4, -6, -8, -10, -12, -13, -15, IFN-alpha, -beta, -gamma, tumor necrosis factor-alpha, -beta, RANTES (regulated on activation normal T-cell express and secreted), MCP-1 (monocyte chemoattractant protein), -2, -3, macrophage inflammatory protein-1 alpha, -1 beta (regulated on activation normal T-cell express and secreted), macrophage colony-stimulating factor, IgG, IgA, and cyclooxygenase 2. All of the specimens were tested as blind replicates, and refrozen plasma was retested 4 months later. To evaluate the reproducibility of the repeat measurements and to examine the correlation between plasma and cervical secretions, we calculated kappa values with 95% confidence intervals among categorized analyte values and Spearman correlation coefficients (rho) among detectable, continuous analyte values. Measurements of all of the analytes in either plasma or cervical secretions were highly reproducible, with all of the kappa > or = 0.78 (70% above 0.90), and all of the rho > or = 0.88 (96% above 0.90). Only IL-1 beta (kappa = 0.60 and rho = 0.82) and IL-6 (kappa = 0.50 and rho = 0.78) levels were strongly correlated between plasma and cervical secretions. IFN-gamma, tumor necrosis factor-beta, RANTES, MCP-1, MCP -2, macrophage inflammatory protein-1 alpha, and macrophage colony-stimulating factor levels were especially poorly correlated between plasma and cervical secretions (kappa < or = 0.25 and rho < or = 0.25). We conclude that recycling immunoaffinity chromatography is a reproducible method of measuring immune profiles from biological specimens, and immune profiles are not well correlated between plasma and cervical secretions, perhaps necessitating cervical collections to study cervix-specific immunity in HPV natural history studies.
Assuntos
Biomarcadores/sangue , Cromatografia/métodos , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/epidemiologia , Adulto , Distribuição por Idade , Idoso , Muco do Colo Uterino/virologia , Quimiocina CCL2/análise , Quimiocina CCL5/análise , Quimiocina CCL8 , Estudos de Coortes , Intervalos de Confiança , Costa Rica/epidemiologia , Feminino , Humanos , Técnicas Imunológicas , Incidência , Programas de Rastreamento/métodos , Pessoa de Meia-Idade , Proteínas Quimioatraentes de Monócitos/análise , Probabilidade , Fatores de Risco , Manejo de Espécimes , Fator de Necrose Tumoral alfa/análiseRESUMO
Eosinophil recruitment into the airways is a feature of asthma in children. However, the mechanisms by which these cells migrate into the airways are not fully understood. The present study investigated the presence of the eosinophil-activating chemokines regulated on activation, normal T-cell expressed and secreted (RANTES), monocyte chemotactic proteins (MCP)-3 and -4, and eotaxins-1 and -2 in the bronchoalveolar lavage (BAL) fluid obtained from both asthmatic (n=10, age 6-10 yrs) and normal children (n=10, age 5-10 yrs). Measurements of chemokines in BAL fluid showed that levels of RANTES, MCPs-3 and -4, and eotaxins-1 and -2 were significantly increased in fluid obtained from asthmatic children when compared with normal children. Among the different chemokines, RANTES was the cytokine released in greatest quantities in BAL fluid from asthmatic children. There was a significant correlation between the concentrations of MCP-4 and eosinophil numbers in BAL fluid and a trend between both chemokines MCP-3 and eotaxin-2 and eosinophils. Interestingly, the levels of most chemokines correlated with one another. These findings suggest that RANTES monocyte chemotactic proteins-3 and -4, and eotaxins-1 and -2 may regulate eosinophil trafficking into the airways of asthmatic children in a coordinated manner.
Assuntos
Asma/metabolismo , Quimiocina CCL5/metabolismo , Quimiocinas CC/metabolismo , Citocinas , Proteínas Quimioatraentes de Monócitos/metabolismo , Asma/patologia , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Quimiocina CCL11 , Quimiocina CCL24 , Quimiocina CCL7 , Criança , Pré-Escolar , Eosinófilos/fisiologia , Feminino , Humanos , Contagem de Leucócitos , MasculinoRESUMO
Several lines of evidence have shown that Trypanosoma cruzi interacts with host extracellular matrix (ECM) components producing breakdown products that play an important role in parasite mobilization and infectivity. Parasite-released antigens also modulate ECM expression that could participate in cell-cell and/or cell-parasite interactions. Increased expression of ECM components has been described in the cardiac tissue of chronic chagasic patients and diverse target tissues including heart, thymus, central nervous system and skeletal muscle of experimentally T. cruzi-infected mice. ECM components may adsorb parasite antigens and cytokines that could contribute to the establishment and perpetuation of inflammation. Furthermore, T. cruzi-infected mammalian cells produce cytokines and chemokines that not only participate in the control of parasitism but also contribute to the establishment of chronic inflammatory lesions in several target tissues and most frequently lead to severe myocarditis. T. cruzi-driven cytokines and chemokines may also modulate VCAM-1 and ICAM-1 adhesion molecules on endothelial cells of target tissues and play a key role in cell recruitment, especially of activated VLA-4+LFA-1+CD8+ T lymphocytes, resulting in a predominance of this cell population in the inflamed heart, central nervous system and skeletal muscle. The VLA-4+-invading cells are surrounded by a fine network of fibronectin that could contribute to cell anchorage, activation and effector functions. Since persistent "danger signals" triggered by the parasite and its antigens are required for the establishment of inflammation and ECM alterations, therapeutic interventions that control parasitism and selectively modulate cell migration improve ECM abnormalities, paving the way for the development of new therapeutic strategies improving the prognosis of T. cruzi-infected individuals