RESUMO
INTRODUCTION: Meningococcal disease is associated with high mortality. When acute kidney injury (AKI) occurs in patients with severe meningococcal disease, it is typically attributable to sepsis, although meningococcal disease and lipopolysaccharide release are rarely investigated. Therefore, we evaluated renal tissue in a mouse model of meningococcal disease. METHODS: Female BALB/c mice were induced to AKI by meningococcal challenge. Markers of renal function were evaluated in infected and control mice. RESULTS: In the infected mice, serum concentrations of tumor necrosis factor alpha, interferon gamma, interleukins (IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-10, and IL-12), and granulocyte-macrophage colony-stimulating factor were elevated, as was renal interstitial infiltration with lymphocytes and neutrophils (p < 0.01 for the latter). Histological analysis showed meningococcal microcolonies in the renal interstitium, without acute tubular necrosis. Infected mice also showed elevated renal expression of toll-like receptor 2, toll-like receptor 4, and Tamm-Horsfall protein. The expression of factors in the intrinsic pathway of apoptosis was equal to or lower than that observed in the control mice. Urinary sodium and potassium were also lower in infected mice, probably due to a tubular defect. CONCLUSION: Our findings corroborate those of other studies of AKI in sepsis. To our knowledge, this is the first time that meningococci have been identified in renal interstitium and that the resulting apoptosis and inflammation have been evaluated. However, additional studies are needed in order to elucidate the mechanisms involved.
Assuntos
Injúria Renal Aguda , Rim , Infecções Meningocócicas , Neisseria meningitidis/isolamento & purificação , Injúria Renal Aguda/sangue , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/imunologia , Injúria Renal Aguda/patologia , Animais , Modelos Animais de Doenças , Perfilação da Expressão Gênica/métodos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Interleucinas/análise , Rim/imunologia , Rim/microbiologia , Rim/patologia , Infecções Meningocócicas/complicações , Infecções Meningocócicas/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Necrose , Infiltração de Neutrófilos , Receptor 2 Toll-Like/análise , Receptor 4 Toll-Like/análise , Uromodulina/análiseRESUMO
INTRODUCTION: The search for more aesthetic and comfortable orthodontic devices has led to an increase in the use of clear aligners. OBJECTIVE: To increase knowledge on biological mechanisms of orthodontic tooth movement using Invisalign aligners. METHODS: This study included 11 patients with a mean age of 23.6 ± 4.8 years. Cases planning included alignment and leveling of lower incisors using Invisalign aligners. Gingival crevicular fluid samples were collected from the lower incisors on the day of delivery of aligner number 1 (T0) and after 1 (T24h), 7 (T7d), and 21 (T21d) days. During the observation period of the study, the patients used only the aligner number 1. Levels of nine cytokines were quantified using Luminex's multi-analysis technology. Non-parametric tests were used for comparisons between cytokine expression levels over time. RESULTS: Cytokine expression levels remained constant after 21 days of orthodontic activation, except those of MIP-1ß, which presented a statistical difference between T24h and T21d with a decrease in the concentration levels. IL-8, GM-CSF, IL-1ß, MIP-1ß, and TNF-α showed the highest concentrations over time. CONCLUSIONS: The different behavior in the levels of the investigated cytokines indicates a role of these biomarkers in the tissue remodeling induced by Invisalign.
Assuntos
Citocinas/análise , Líquido do Sulco Gengival/química , Técnicas de Movimentação Dentária , Quimiocina CCL2/análise , Quimiocina CCL4/análise , Fatores Estimuladores de Colônias/análise , Citocinas/metabolismo , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Humanos , Incisivo , Interleucina-17/análise , Interleucina-1beta/análise , Interleucina-7/análise , Interleucina-8/análise , Masculino , Aparelhos Ortodônticos Removíveis , Fator de Necrose Tumoral alfa/análise , Adulto JovemRESUMO
ABSTRACT Introduction: The search for more aesthetic and comfortable orthodontic devices has led to an increase in the use of clear aligners. Objective: To increase knowledge on biological mechanisms of orthodontic tooth movement using Invisalign aligners. Methods: This study included 11 patients with a mean age of 23.6 ± 4.8 years. Cases planning included alignment and leveling of lower incisors using Invisalign aligners. Gingival crevicular fluid samples were collected from the lower incisors on the day of delivery of aligner number 1 (T0) and after 1 (T24h), 7 (T7d), and 21 (T21d) days. During the observation period of the study, the patients used only the aligner number 1. Levels of nine cytokines were quantified using Luminex's multi-analysis technology. Non-parametric tests were used for comparisons between cytokine expression levels over time. Results: Cytokine expression levels remained constant after 21 days of orthodontic activation, except those of MIP-1β, which presented a statistical difference between T24h and T21d with a decrease in the concentration levels. IL-8, GM-CSF, IL-1β, MIP-1β, and TNF-α showed the highest concentrations over time. Conclusions: The different behavior in the levels of the investigated cytokines indicates a role of these biomarkers in the tissue remodeling induced by Invisalign.
RESUMO Introdução: a busca por dispositivos ortodônticos mais estéticos e confortáveis gerou um aumento no uso de alinhadores transparentes. Objetivo: ampliar o conhecimento sobre os mecanismos biológicos associados ao movimento dentário ortodôntico promovido por alinhadores Invisalign®. Métodos: a amostra foi constituída por 11 pacientes, com idade média de 23,6 ± 4,8 anos. O planejamento dos casos incluiu alinhamento e nivelamento de incisivos inferiores usando os alinhadores. O fluido gengival crevicular foi coletado na superfície vestibular de incisivos inferiores no dia da entrega do alinhador número 1 (T0) e após 1 (T24h), 7 (T7d) e 21 (T21d) dias. Durante o período de observação do estudo, os pacientes utilizaram apenas o alinhador número 1. Os níveis de nove citocinas foram quantificados por meio do sistema Luminex de multianálise. Testes não paramétricos foram realizados para comparações entre os níveis de expressão de citocinas ao longo do tempo. Resultados: a concentração das citocinas manteve-se constante após 21 dias de ativação ortodôntica, exceto a MIP-1β, que apresentou uma redução estatisticamente significativa entre os tempos T24h e T21d. As IL-8, GM-CSF, IL-1β, MIP-1β e TNF-α apresentaram as maiores concentrações ao longo do tempo. Conclusão: a constância na expressão dos níveis das citocinas parece estar compatível com o estímulo mecânico induzido por alinhadores.
Assuntos
Humanos , Masculino , Feminino , Adulto Jovem , Técnicas de Movimentação Dentária , Citocinas/análise , Líquido do Sulco Gengival/química , Aparelhos Ortodônticos Removíveis , Citocinas/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Interleucina-8/análise , Fatores Estimuladores de Colônias/análise , Interleucina-7/análise , Fator de Necrose Tumoral alfa/análise , Quimiocina CCL2/análise , Interleucina-17/análise , Interleucina-1beta/análise , Quimiocina CCL4/análise , IncisivoRESUMO
ABSTRACT It is well established that protein malnutrition (PM) impairs immune defenses and increases susceptibility to infection. Macrophages are cells that play a central role in innate immunity, constituting one of the first barriers against infections. Macrophages produce several soluble factors, including cytokines and growth factors, important to the immune response. Among those growth factors, granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF). GM-CSF and M-CSF are important to monocyte and macrophage development and stimulation of the immune response process. Knowing the importance of GM-CSF and M-CSF, we sought to investigate the influence of PM on macrophage production of these growth factors. Two-month-old male BALB/c mice were subjected to PM with a low-protein diet (2%) and compared to a control diet (12%) mouse group. Nutritional status, hemogram and the number of peritoneal cells were evaluated. Additionally, peritoneal macrophages were cultured and the production of GM-CSF and M-CSF and mRNA expression were evaluated. To determine if PM altered macrophage production of GM-CSF and M-CSF, they were stimulated with TNF-α. The PM animals had anemia, leukopenia and a reduced number of peritoneal cells. The production of M-CSF was not different between groups; however, cells from PM animals, stimulated with or without TNF-α, presented reduced capability to produce GM-CSF. These data imply that PM interferes with the production of GM-CSF, and consequently would affect the production and maturation of hematopoietic cells and the immune response.
Assuntos
Ratos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Fator Estimulador de Colônias de Macrófagos/análiseRESUMO
AIM: The aim of this study was to evaluate the levels of a wide panel of cyto/chemokines in the gingival crevicular fluid (GCF) of uncontrolled type 2 diabetic subjects as compared with non-diabetic subjects with periodontitis. METHODS: Twenty-six uncontrolled type 2 diabetic subjects (glycated haemoglobin levels >7.5%) and 20 non-diabetic subjects with chronic periodontitis were enrolled in this study. The levels of 14 cyto/chemokines were measured in the GCF of healthy and diseased sites of the diabetic and non-diabetic subjects using multiplex bead immunoassays. RESULTS: The concentrations of eotaxin, macrophage inflammatory protein-1α, granulocyte-macrophage colony-stimulating factor, interleukin (IL)-6, tumour necrosis factor-α and IL-12 were higher in healthy and diseased sites of diabetic than non-diabetic subjects, after adjustment for multiple comparisons (p < 0.0035). CONCLUSION: Uncontrolled type 2 diabetes mellitus modulated the local levels of several cyto/chemokines at both healthy and diseased periodontal sites in favour of a proinflammatory profile, which may partially explain the greater susceptibility of diabetic subjects to periodontal breakdown.
Assuntos
Periodontite Crônica/imunologia , Diabetes Mellitus Tipo 2/imunologia , Líquido do Sulco Gengival/imunologia , Mediadores da Inflamação/análise , Adulto , Glicemia/análise , Quimiocina CCL3/análise , Quimiocinas/análise , Quimiocinas CC/análise , Periodontite Crônica/complicações , Citocinas/análise , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/complicações , Feminino , Líquido do Sulco Gengival/química , Hemoglobinas Glicadas/análise , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Humanos , Interleucina-12/análise , Interleucina-6/análise , Masculino , Pessoa de Meia-Idade , Fator de Necrose Tumoral alfa/análiseRESUMO
AIM: To investigate the effect of photodynamic therapy (PDT) as adjunct to mechanical therapy in furcations. MATERIALS AND METHODS: A double-blind, parallel, randomized controlled clinical trial was conducted in subjects presenting class II furcations. The subjects were randomly allocated to a test (PDT; n = 16) or control group (non-activated laser/only photosensitizer; n = 21). At baseline, 3 and 6 months, clinical, microbiological and cytokine pattern evaluation was performed. Clinical attachment level was defined as the primary outcome variable. RESULTS: Clinical parameters improved after both therapies (p < 0.05) with no differences between groups at any time point (p > 0.05). At 6 months, real-time PCR evaluation showed a decrease in Porphyromonas gingivalis and Tannerella forsythia only in the PDT group (p < 0.05) with no inter-group differences. Regarding cytokines, IL-4 and IL-10 levels increased in both groups at 6 months. GM-CSF, IL-8, IL-1ß and IL-6 levels decreased only in the PDT group after 3 months (p < 0.05). At 3 months, inter-group analyses showed that GM-CSF, IFN-γ, IL-6 and IL-8 levels were lower in the PDT group. At 6 months, lower IL-1ß levels were also observed in the PDT group (p < 0.05). CONCLUSION: Photodynamic therapy did not promote clinical benefits for class II furcations; however, advantages in local levels of cytokines and a reduction in periodontopathogens were demonstrated.
Assuntos
Defeitos da Furca/tratamento farmacológico , Fotoquimioterapia/métodos , Aggregatibacter actinomycetemcomitans/efeitos dos fármacos , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Carga Bacteriana/efeitos dos fármacos , Bacteroides/efeitos dos fármacos , Bacteroides/isolamento & purificação , Periodontite Crônica/tratamento farmacológico , Periodontite Crônica/microbiologia , Terapia Combinada , Raspagem Dentária/métodos , Método Duplo-Cego , Feminino , Seguimentos , Defeitos da Furca/classificação , Defeitos da Furca/microbiologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Humanos , Interferon gama/análise , Interleucina-10/análise , Interleucina-1beta/análise , Interleucina-4/análise , Interleucina-6/análise , Interleucina-8/análise , Lasers Semicondutores/uso terapêutico , Masculino , Pessoa de Meia-Idade , Perda da Inserção Periodontal/tratamento farmacológico , Perda da Inserção Periodontal/microbiologia , Fármacos Fotossensibilizantes/uso terapêutico , Porphyromonas gingivalis/efeitos dos fármacos , Porphyromonas gingivalis/isolamento & purificação , Estudos Prospectivos , Aplainamento Radicular/métodos , Resultado do TratamentoRESUMO
A stability-indicating capillary zone electrophoresis (CZE) method was validated for the analysis of granulocyte-macrophage colony-stimulating factor (rhGM-CSF) using leuprorelin acetate (LA), as internal standard (IS). A fused-silica capillary (75 µm i.d.; effective length, 72 cm) was used at 25 °C; the applied voltage was 12 kV. The background electrolyte solution consisted of 50mM di-sodium hydrogen phosphate solution at pH 8.8. Injections were performed using a pressure mode at 50 mbar for 9s, with detection by photodiode array detector set at 200 nm. Specificity and stability-indicating capability were established in degradation studies, which also showed that there was no interference of the excipients. The method was linear over the concentration range of 2.5-200 µg mL(-1) (r(2)=0.9995) and the limit of detection (LOD) and limit of quantitation (LOQ) were 0.79 µg mL(-1) and 2.5 µg mL(-1), respectively. The accuracy was 99.14% with bias lower than 1.40%. The method was applied to the quantitative analysis of biopharmaceutical formulations, and the results were correlated to those of a validated reversed-phase LC method (RP-LC), and an in vitro bioassay, showing non-significant differences (p>0.05).
Assuntos
Eletroforese Capilar/métodos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Proteínas Recombinantes/análise , Bioensaio , Calibragem , Linhagem Celular , Cromatografia de Fase Reversa , Humanos , Concentração de Íons de Hidrogênio , Leuprolida/análise , Limite de Detecção , Fosfatos/química , Dióxido de Silício/químicaRESUMO
OBJECTIVE: The aim of this study was to investigate the effects of PRP on SAOS-2 cells in terms of cytokine expression, cell activity and oxidative stress. DESIGN: Cell line SAOS-2 (1×10(5)cells/mL) were grown in culture medium α-MEM with 10% FBS for 24h and stimulated (or not) with PRP at concentrations of 3, 10 and 20%, LPS (E. coli, 10g/mL) and IL-1ß (1mg/mL) for 24h. The supernatant was collected and analyzed for the expression of cytokines in a panel array, ALP using a commercial kit and NO(2)(-) with Griess reaction method. Also, the cells were analyzed using Western blot for RANKL and slot blotting for nitrotyrosine expression. RESULT: There were no significant differences amongst the groups in terms of NO(2)(-), protein nitrotyrosine content and RANKL expression. However, all stimuli increased ALP activity and in case of PRP, it was in a dose-dependent manner (p<0.001). Also, all stimuli induced an increase in cytokines and chemokines expression, but only PRP promoted an increase of component C5, sICAM-1 and RANTES expression. Whilst IL-1 receptor antagonist (IL-1ra) expression was down-regulated by PRP, both LPS and IL-1ß caused up-regulation of this cytokine. CONCLUSIONS: PRP can stimulate osteoblast activity and cytokine/chemokine release, as well as indicate some of the mediators that can (and cannot) be involved in this activation.
Assuntos
Fosfatase Alcalina/análise , Citocinas/análise , Osteoblastos/metabolismo , Plasma Rico em Plaquetas/fisiologia , Linhagem Celular Tumoral , Quimiocina CCL5/análise , Quimiocina CXCL1/análise , Complemento C5/análise , Relação Dose-Resposta a Droga , Escherichia coli , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Humanos , Molécula 1 de Adesão Intercelular/análise , Proteína Antagonista do Receptor de Interleucina 1/análise , Interleucina-1beta/farmacologia , Interleucinas/análise , Lipopolissacarídeos/farmacologia , Óxido Nítrico/análise , Osteoblastos/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Ligante RANK/análise , Tirosina/análogos & derivados , Tirosina/análiseRESUMO
AIM: To examine changes in levels of gingival crevicular fluid (GCF) cytokines, after periodontal therapy of generalized aggressive periodontitis (GAgP). MATERIALS AND METHODS: Twenty-five periodontally healthy and 24 GAgP subjects had periodontal clinical parameters measured and gingival crevicular fluid (GCF) samples collected from up to 14 sites/subject. GCF samples were analysed using multiplex bead immunoassay for: GM-CSF, IFN-γ, IL-10, IL-1ß, IL-2, IL-6 and TNF-α. Aggressive periodontitis subjects were randomly assigned to either scaling and root planing (SRP) alone or SRP plus systemic amoxicillin (500 mg) and metronidazole (400 mg) 3 times a day for 14 days. Clinical parameters and GCF cytokines were re-measured 6 months after treatment. Differences over time were analysed using the Wilcoxon test and between groups using the Mann-Whitney test. RESULTS: Significant reductions in GCF GM-CSF, IL-1ß and the ratio IL-1ß/IL-10 and increases in GCF IL-6 were detected after therapy. The mean change in GCF cytokines did not differ significantly between groups. CONCLUSIONS: Periodontal therapy improved GCF cytokine profiles by lowering IL-1ß and increasing IL-10 levels. The reduction in GCF GM-CSF after therapy implicates this cytokine in the pathogenesis of GAgP. There was no difference between therapies in changes of GCF cytokines.
Assuntos
Periodontite Agressiva/metabolismo , Periodontite Agressiva/terapia , Antibacterianos/uso terapêutico , Citocinas/metabolismo , Raspagem Dentária , Líquido do Sulco Gengival/química , Adulto , Periodontite Agressiva/patologia , Amoxicilina/uso terapêutico , Análise de Variância , Citocinas/análise , Método Duplo-Cego , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Interferon gama/análise , Interferon gama/metabolismo , Interleucina-10/análise , Interleucina-10/metabolismo , Interleucina-1beta/análise , Interleucina-2/análise , Interleucina-2/metabolismo , Interleucina-6/análise , Interleucina-6/metabolismo , Masculino , Metronidazol/uso terapêutico , Estatísticas não Paramétricas , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/metabolismo , Adulto JovemRESUMO
Six anti-E. coli rhGM-CSF monoclonal antibodies were generated using hybridoma technology. One of these showed identical affinity for the CHO and E. coli-derived cytokine (dissociation constants of 0.14 +/- 0.01 nM and 0.13 +/- 0.01 nM, respectively), mapping an epitope that is not hindered by carbohydrates. The antibody was used to develop a simple, specific and sensitive competitive ELISA to quantify the entire set of rhGM-CSF glycoforms (detection limit of 780 pg/ml) and it was successful as an affinity ligand to purify them. Therefore, this particular antibody is a useful, reliable and reagent for most immunochemical purposes with the aim of detecting, quantifying and purifying the highly heterogeneous collection of the CHO-derived rhGM-CSF isoforms.
Assuntos
Anticorpos Monoclonais/análise , Formação de Anticorpos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Processamento de Proteína Pós-Traducional/imunologia , Animais , Anticorpos Monoclonais/metabolismo , Afinidade de Anticorpos , Células CHO , Cricetinae , Cricetulus , Ensaio de Imunoadsorção Enzimática , Mapeamento de Epitopos , Escherichia coli , Feminino , Glicosilação , Fator Estimulador de Colônias de Granulócitos e Macrófagos/química , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/análise , Proteínas Recombinantes/química , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismoRESUMO
Advanced complicated atherosclerotic lesions have been related to many factors, including inflammation, infectious agents, and growth factors. Mycoplasma pneumoniae (MP) and Chlamydia pneumoniae (CP), inflammation, and growth factors have been associated with severe atherosclerotic lesions in necropsy material in recent work at our lab. The present study intends to clarify the pathogenesis of atherosclerosis, analyzing which of these elements (macrophages, MP, CP, lymphocytes, and growth factors) are associated with initial development of atherosclerotic lesions, discriminating elements related to stabilization of the plaque versus those related to subendothelial active accumulation of macrophages in living patients. Surgical ascending aorta fragments presenting mild atherosclerotic lesions from 30 coronary atherosclerotic patients were immunohistochemically quantified regarding CP, MP, T cells (CD4, CD8), B cells (CD20), macrophages (CD68), and growth factors [platelet-derived growth factor A (PDGF-A), PDGF-B, transforming growth factor-beta (TGF-beta), granulocyte-macrophage colony-stimulating factor (GM-CSF)]. Cases were grouped according to the presence or not of active accumulation of macrophages at the subendothelium that indicates atheroma in development: group I (GI) fragments with <4 CD68+ cells/x400 field, in normal distribution (mean 1.8 +/- 1) representing stable atherosclerotic mild lesion, and GII fragments presenting >or=4 CD68+ cells/x400 field, in a non-normal distribution, mean (8.9 +/- 4.8, atheromas in progress), which was followed by increased number of lymphocytes. The median number in GI was significantly lower than that in GII: CD4 T (2.5 vs. 7.7), CD8 T (1.0 vs. 5.5), and CD20 B (1.5 vs. 5.5) cells/x400 field, p < 0.001. Percentage area positive for CP antigens was significantly lower in GI than in GII: 1.0 vs. 9.2, p < 0.001. There was a higher percentage area occupied by MP than CP in both GI and GII (7.8 vs. 13.8). There was no difference regarding mean number of growth factor-positive cells/x400 field: PDGF-A, 1.4 vs. 3.9; PDGF-B, 3.4 vs. 5.7; TGF-beta, 0.9 vs. 2.2; and GM-CSF, 2.0 vs. 2.2. Considering all cases, a positive correlation was seen between inflammatory cells and CP+ cells (r > 0.5 and p < 0.01). Growth factors did not correlate with inflammatory cells, CP, or MP and were usually seen in smooth muscle cell and fibrotic areas. Study of initial atherosclerotic lesions showed that MP is present in both kinds of lesion: stable and active subendothelial accumulation of macrophages. Stabilization was related to proportional increase of both infectious agents, which were also related to increased amount of PDGF-A and PDGF-B. Active macrophage accumulation lesions were related to higher elevation in CP concentration at subendothelial regions, in association with B cells, but not of MP and growth factors. MP and CP, inflammation, and growth factors, which were already described in severe atherosclerotic lesions in necropsy material, are also present in mild lesions in living patients, strongly favoring a pathogenetic role for these bacteria in the pathogenesis of atherosclerosis. Predominance of CP in relation to MP may favor progression of the plaque, which is associated with increased B-cell proliferation. PDGF-A and PDGF-B are associated with plaque stability, at least in arterial segments not prone for development of complicated lesions.
Assuntos
Aorta Torácica/patologia , Doenças da Aorta/patologia , Aterosclerose/patologia , Citocinas/análise , Bactérias Gram-Negativas/isolamento & purificação , Peptídeos e Proteínas de Sinalização Intercelular/análise , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos CD/análise , Antígenos CD20/análise , Antígenos de Diferenciação Mielomonocítica/análise , Aorta Torácica/química , Aorta Torácica/imunologia , Aorta Torácica/microbiologia , Doenças da Aorta/imunologia , Doenças da Aorta/metabolismo , Doenças da Aorta/microbiologia , Aterosclerose/imunologia , Aterosclerose/metabolismo , Aterosclerose/microbiologia , Linfócitos B/patologia , Biópsia , Chlamydophila pneumoniae/isolamento & purificação , Progressão da Doença , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Humanos , Inflamação/patologia , Macrófagos/patologia , Masculino , Pessoa de Meia-Idade , Mycoplasma pneumoniae/isolamento & purificação , Fator de Crescimento Derivado de Plaquetas/análise , Linfócitos T/patologia , Fator de Crescimento Transformador beta/análiseRESUMO
BACKGROUND, AIMS: Neutrophil cells constitute the first defense barrier against the oral bacterial challenge in the periodontium. Reduction of neutrophils could impair this response against periopathogenic bacteria such as Porphyromonas gingivalis. Our previous work implicates the apoptosis of neutrophils in the pathogenesis of periodontitis. We now demonstrate that granulocyte monocyte-colony stimulating factor (GM-CSF) present in the gingival crevicular fluid (GCF) and secreted during the immune response reduces the apoptosis of neutrophils. METHOD: In this study, the presence of GM-CSF and tumor necrosis factor-alpha (TNF-alpha) in GCF was determined in samples obtained from adult patients with periodontitis and from control subjects with clinically healthy gingiva. GCF was collected for 30 s using Periopaper(R) strips, and cytokines were quantified by ELISA. We used ex vivo culture of gingival tissue biopsies for 2 and 4 days in the presence of GM-CSF. Apoptosis was determined using the terminal TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique, and expression of Bax by immunohistochemistry. RESULTS: The presence of GM-CSF and TNF-alpha was detected in the majority of sites from periodontal patients (83.3% and 63.3%, respectively), presenting a total amount of 27.65 and 42.38 pg, respectively. GM-CSF reduces the neutrophil apoptosis determined by double staining with TUNEL and myeloperoxidase and by a reduction of Bax expression. CONCLUSIONS: These findings suggest a novel mechanism by which neutrophils specifically accumulate in adult patients with periodontitis.
Assuntos
Apoptose , Fator Estimulador de Colônias de Granulócitos e Macrófagos/fisiologia , Neutrófilos/fisiologia , Periodontite/imunologia , Proteínas Proto-Oncogênicas c-bcl-2 , Fator de Necrose Tumoral alfa/fisiologia , Adulto , Estudos de Casos e Controles , Doença Crônica , Feminino , Expressão Gênica , Líquido do Sulco Gengival/química , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Humanos , Técnicas Imunoenzimáticas , Marcação In Situ das Extremidades Cortadas , Masculino , Pessoa de Meia-Idade , Periodontite/fisiopatologia , Proteínas Proto-Oncogênicas/análise , Proteínas Proto-Oncogênicas/biossíntese , Fator de Necrose Tumoral alfa/análise , Proteína X Associada a bcl-2RESUMO
Cytokines released by immune system cells play an important role in cyst enlargement. This study aimed to determine, by ELISA, the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin 3 (IL-3), and IL-6 in fluid and tissue from human radicular cysts. GM-CSF was found in 42.8% of the fluid samples (164.3 pg/mL) and IL-6 in 92.8% (641.4 pg/mL). No IL-3 was detected in any fluid samples. In the tissue samples, 28.6% were positive for IL-3 (369.2 pg/mL), 86.4% for IL-6 (92.4 pg/mL), and 95.8% for GM-CSF (200.5 pg/mL). It can be concluded that GM-CSF and IL-6 were widely found in the fluid and tissue samples. In contrast, IL-3 was found only in the cystic tissue, even though in few lesions. These cytokines may contribute to the inflammation, cystic growth, and bone resorption that characterize cystic lesions.
Assuntos
Líquido Cístico/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Interleucina-3/análise , Interleucina-6/análise , Cisto Radicular/imunologia , Perda do Osso Alveolar/imunologia , Líquido Cístico/química , Ensaio de Imunoadsorção Enzimática , Humanos , Cisto Radicular/química , Estatísticas não ParamétricasRESUMO
Detectamos as citocinas interleucina-6 (IL-6) e granulócito-macrófago-CSF (GM-CSF) por ELISA no LCR e soro de 30 pacientes infectados por HIV classificados como tendo AIDS-demência complexo (ACD) e de 20 pacientes com outras doenças neurológicas (OND). Encontramos elevada incidência de IL-C e GM-CSF detectável no LCR de pacientes com ADC, em relaçäo aos pacientes com OND. Diferenças estatísticas näo foram observadas entre os dois grupos de pacientes para níveis de IL-6 e GM-CSF no soro. Esses resultados sugerem síntese intratectal dessas citocinas e sua possível participaçäo na patogenia da ADC
Assuntos
Humanos , Masculino , Feminino , Adulto , Complexo AIDS Demência/líquido cefalorraquidiano , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Interleucina-6/análise , Complexo AIDS Demência/sangue , Doenças do Sistema Nervoso Central/líquido cefalorraquidiano , Doenças do Sistema Nervoso Central/sangue , Ensaio de Imunoadsorção EnzimáticaRESUMO
The present results demonstrate that macrophages from mice susceptible to infection with Leishmania mexicana amazonensis sustain a higher production of granulocyte-macrophage colony-stimulating factor (GM-CSF) throughout the in vitro infection than macrophages from a resistant strain. Resident macrophages from BALB/c and C57B1/10 mice were infected with promastigotes of L. mexicana amazonensis and the amount of biologically active GM-CSF was measured in the supernatants collected at different times of infection. Measurements were made by bone marrow and GM-CSF/interleukin-3 addicted cell proliferation. Because GM-CSF is a disease-exacerbating cytokine, its differential production by infected macrophages may be one of the mechanisms defining resistance or susceptibility to a leishmanial infection.