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1.
Am J Physiol Lung Cell Mol Physiol ; 320(2): L241-L245, 2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-33146565

RESUMO

Microvesicles (MVs) released from almost all cells are recognized as cell communication tools. MVs have been investigated in several inflammatory diseases but poorly in biological fluids like bronchoalveolar lavage (BAL) of smokers. The purpose of this study was to investigate the presence and source of MVs in BAL of smokers with and without chronic obstructive pulmonary disease (COPD) compared with nonsmoking controls. Using flow cytometry in BAL, we detected endothelial and alveolar macrophage (AM)-derived MVs and found a higher number of AM-MVs in the BAL of smokers with COPD than in smokers without COPD and nonsmokers, which correlated with the pack-years (r = 0.46; P = 0.05) and with the degree of airway obstruction measured by the forced expiratory volume in 1 s percent predicted (r = -0.56; P = 0.01). Endothelial and alveolar macrophage-derived MVs are present and measurable in human BAL fluid. In response to smoking and to the development of COPD, inflammatory signals in AM-derived MVs can be quantified, and their numbers are related to the pack-years and the decrease in lung function. These results open the opportunity for future investigation of these microvesicles as biomarkers and possible mechanistic guides in COPD.


Assuntos
Biomarcadores/análise , Líquido da Lavagem Broncoalveolar/química , Micropartículas Derivadas de Células/patologia , Macrófagos Alveolares/patologia , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Fumar/efeitos adversos , Idoso , Feminino , Volume Expiratório Forçado , Humanos , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/etiologia , Testes de Função Respiratória
2.
Methods Mol Biol ; 1884: 87-117, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30465197

RESUMO

This chapter will describe the current methodologies to isolate and expand NK cells from Peripheral Blood (PB) or tissues for "in vitro" studies, including NK cell antitumor immune function. In addition, methods to induce NK cell maturation, differentiation, and expansion from CD34+ precursors will also be described. Finally, it will also be treated the topical issue of the characterization of new functionally and phenotypically defined NK cell subsets.


Assuntos
Separação Celular/métodos , Citometria de Fluxo/métodos , Células Matadoras Naturais/imunologia , Subpopulações de Linfócitos/imunologia , Neoplasias/imunologia , Antígenos CD34/metabolismo , Diferenciação Celular/imunologia , Separação Celular/instrumentação , Células Cultivadas , Citocinas/imunologia , Citocinas/metabolismo , Sangue Fetal/citologia , Citometria de Fluxo/instrumentação , Corantes Fluorescentes/química , Humanos , Vigilância Imunológica , Células Matadoras Naturais/metabolismo , Subpopulações de Linfócitos/metabolismo , Células Precursoras de Linfócitos T/fisiologia , Cultura Primária de Células/instrumentação , Cultura Primária de Células/métodos
3.
Environ Toxicol Pharmacol ; 39(1): 93-101, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25473821

RESUMO

The aim was to assess the individual susceptibility to mitochondrial impairment induced by ex vivo exposure to vanadium, an airborne pro-oxidant pollutant. In lymphocyte cultures V(IV)-treated of forty-five healthy subjects, we evaluated the mitochondrial transmembrane potential (Δψm) and the H2O2 in comparison to background values. As variables, we included both lifestyle factors and genetic polymorphisms (GSTM1 and GSTT1 variants, and C677T and A1298C variants of methylenetetrahydrofolate reductase MTHFR). H2O2 mitochondrial content increased significantly (P<0.05) after metal exposure while, in comparison to basal Δψm, both depolarisation and hyperpolarisation were recorded. This underlined the mitochondrial dysfunction vanadium-induced that worsens the redox imbalance by endogenous ROS overproduction. Only age was found to contribute significantly to the high inter-individual variability, as assessed by multivariate analysis. In older subjects, the H2O2/Δψm values underline the organelle impairment and, under V-exposure, Δψm values were inversely related to age (R=-0.591; P=0.012).


Assuntos
Poluentes Ambientais/toxicidade , Peróxido de Hidrogênio/metabolismo , Linfócitos/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Vanádio/toxicidade , Adulto , Fatores Etários , Células Cultivadas , Feminino , Genótipo , Glutationa Transferase/genética , Humanos , Linfócitos/metabolismo , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Pessoa de Meia-Idade , Mitocôndrias/metabolismo , Polimorfismo Genético
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