In order to study the signal pathway secreting type Ⅰ interferon in porcine alveolar macrophages (PAMs) infected with porcine circovirus type 2 (PCV2), the protein and the mRNA expression levels of cGAS/STING pathways were analyzed by ELISA, Western blotting and quantitative reverse transcriptase PCR in PAMs infected with PCV2. In addition, the roles of cGAS, STING, TBK1 and NF-κB/P65 in the generation of type I interferon (IFN-I) from PAMs were analyzed by using the cGAS and STING specific siRNA, inhibitors BX795 and BAY 11-7082. The results showed that the expression levels of IFN-I increased significantly at 48 h after infection with PCV2 (P<0.05), the mRNA expression levels of cGAS increased significantly at 48 h and 72 h after infection (P<0.01), the mRNA expression levels of STING increased significantly at 72 h after infection (P<0.01), and the mRNA expression levels of TBK1 and IRF3 increased at 48 h after infection (P<0.01). The protein expression levels of STING, TBK1 and IRF3 in PAMs infected with PCV2 were increased, the content of NF-κB/p65 was decreased, and the nuclear entry of NF-κB/p65 and IRF3 was promoted. After knocking down cGAS or STING expression by siRNA, the expression level of IFN-I was significantly decreased after PCV2 infection for 48 h (P<0.01). BX795 and BAY 11-7082 inhibitors were used to inhibit the expression of IRF3 and NF-κB, the concentration of IFN-I in BX795-treated group was significantly reduced than that of the PCV2 group (P<0.01), while no significant difference was observed between the BAY 11-7028 group and the PCV2 group. The results showed that PAMs infected with PCV2 induced IFN-I secretion through the cGAS/STING/TBK1/IRF3 signaling pathway.
Subject(s)Animals , Cells, Cultured , Circovirus , Interferon Type I/genetics , Macrophages, Alveolar/virology , Membrane Proteins/metabolism , Nucleotidyltransferases/metabolism , Signal Transduction , Swine
The aim of this study was to investigate the effects of polarization program on the ability of macrophages to regulate iron metabolism. M1 and M2 macrophages were propagated in vitro from porcine alveolar macrophages 3D4/2 and polarized by cytokines. The 3D4/2 macrophages were treated with 20 ng/mL interferon gamma (IFN-γ) and 10 ng/mL interleukin-4 (IL-4) combined with 10 ng/mL macrophage colony-stimulating factor (M-CSF) to induce polarization to M1 and M2, respectively. After incubation for 24 h, the expression levels of inflammatory factors and iron-metabolism genes were determined using real-time qPCR, Western bot and immunofluorescence. The M1/M2 macrophages culture media supernatant was collected and used to treat porcine intestinal epithelial cells IPEC-J2. The proliferation ability of IPEC-J2 was detected using CCK-8 assay kit. Following exogenous addition of ammonium ferric citrate (FAC) to M1/M2 macrophages, the phagocytic function of macrophages was detected using fluorescein isothiocyanate-dextran (FITC-dextran) and flow cytometry. The results showed that, compared with control, M1 macrophages had higher mRNA levels of iron storage proteins (ferritin heavy and light polypeptide, i.e. FtH and FtL), hepcidin and lipocalin-2, as well as iron content. Moreover, iron enhanced the ability of M1 macrophages to phagocytize FITC-dextran. There was no significant change in these mRNA expression levels in M2 macrophages, but the mRNA expression levels of ferroportin and transferrin receptor were up-regulated. In addition, the conditioned media supernatant from M2 macrophages promoted cell proliferation of IPEC-J2. These findings indicate that M1 macrophages tend to lock iron in the cell and reduce extracellular iron content, thereby inhibiting the proliferation of extracellular bacteria. While M2 macrophages tend to excrete iron, which contributes to the proliferation of surrounding cells and thus promotes tissue repair.
Subject(s)Animals , Cytokines , Ferritins , Iron/metabolism , Macrophages/metabolism , Macrophages, Alveolar/metabolism , Swine
Alveolar macrophages (AMs) are an essential part of defense mechanisms within the lungs and their phagocytic activity is important for organ homeostasis. The phagocytic ability of AMs obtained from bronchoalveolar lavage from 17 mature mixed-breed pleasure horses (8 healthy and 9 diagnosed with mild equine asthma) was studied through assays with Leishmania (Viannia) braziliensis promastigotes, which enabled the calculation of a phagocytic index (PI) and a survival index (SI). Results indicate that phagocytic activity of AMs in asthma affected horses is similar to healthy horses, while leishmanicidal activity is significantly increased in horses with asthma.(AU)
Os macrófagos alveolares (MAs) são uma parte essencial dos mecanismos de defesa dentro dos pulmões e sua atividade fagocítica é importante para a homeostase desse órgão. A capacidade fagocitária dos MAs obtidos do lavado broncoalveolar de 17 equinos adultos, sem raça definida (oito saudáveis e nove com diagnóstico de asma equina leve), foi estudada por meio de ensaios com promastigotas de Leishmania (Viannia) braziliensis. Foi calculado o índice fagocítico e o índice de sobrevivência. Os resultados indicam que a atividade fagocítica de MAs em cavalos com asma é semelhante a cavalos saudáveis, enquanto a atividade leishmanicida está significativamente aumentada em cavalos com essa enfermidade.(AU)
Subject(s)Animals , Asthma/veterinary , Leishmania braziliensis , Macrophages, Alveolar/parasitology , Horses/parasitology , Phagocytosis
The aim of the present study was to investigate the effect of salidroside (Sal) on inflammatory activation induced by lipopolysaccharide (LPS) in the co-culture of rat alveolar macrophages (AM) NR 8383 and type II alveolar epithelial cells (AEC II) RLE-6TN. CCK-8 colorimetric method was used to detect cell proliferation percentage. The enzyme-linked immunosorbent assay (ELISA) was used to determine the content of tumor necrosis factor alpha (TNF-α), macrophage inflammatory protein-2 (MIP-2) and interleukin-10 (IL-10) in the supernatant. Western blot was used to examine the expression levels of phosphorylated AKT (p-AKT) and total AKT protein. The results showed that pretreatment of RLE-6TN cells or co-culture of RLE-6TN and NR 8383 cells with 32 and 128 µg/mL Sal for 1 h, followed by continuous culture for 24 h, significantly increased the cell proliferation (P < 0.05). Compared with control group, 32 and 128 µg/mL Sal pretreatment significantly increased the ratio of p-AKT/AKT in RLE-6TN cells (P < 0.05). Pretreatment of 32 µg/mL Sal not only inhibited the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05), but also enhanced the inhibitory effect of RLE-6TN and NR 8383 cells co-culture on the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05). In addition, 32 µg/mL Sal pretreatment promoted LPS-induced IL-10 secretion by NR 8383 cells (P < 0.05), and enhanced the promoting effect of co-culture of RLE-6TN and NR 8383 cells on the IL-10 secretion by LPS-induced NR 8383 cells (P < 0.05). In conclusion, Sal may directly inhibit LPS-induced inflammatory activation of AM (NR 8383), promote the proliferation of AEC II (RLE-6TN) through PI3K/AKT signaling pathway, and enhance the regulatory effect of AEC II on LPS-induced inflammatory activation of AM.
Subject(s)Alveolar Epithelial Cells , Metabolism , Animals , Cell Line , Chemokine CXCL2 , Metabolism , Coculture Techniques , Glucosides , Pharmacology , Interleukin-10 , Metabolism , Lipopolysaccharides , Macrophages, Alveolar , Metabolism , Phenols , Pharmacology , Phosphatidylinositol 3-Kinases , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Rats , Signal Transduction , Tumor Necrosis Factor-alpha , Metabolism
OBJECTIVE@#To investigate the effect of sulfur dioxide (SO2) on the apoptosis of alveolar macrophage (AM) in lung protection of limb ischemia/reperfusion (I/R) induced acute lung injury (ALI), and to find a new target for the control of inflammatory response.@*METHODS@#Twenty pathogen-free, adult male Sprague-Dawley (SD) rats (180-230 g) were used in this study. Five rats were to be used for limb ischemia/reperfusion, then plasma was extracted as ischemia/reperfusion serum stimulation. Fifteen rats were to be used for extracting AM by bronchoalveolar lavage. The AM was isolated and cultured, then the cell count was adjusted to 1×106/mL, and randomly divided into the following 4 groups (n=6): control group, I/R group, SO2 group, and I/R+SO2 group. The I/R group was given ischemia/reperfusion serum (500 μg/L) to stimulate 6 h; the SO2 group was given an SO2 donor, Na2SO3/NaHSO3 [(0.54 mmol/kg) / (0.18 mmol/kg)]; and the I/R+SO2 group was given the same ischemia/reperfusion serum and Na2SO3/NaHSO3 at the same time. The level of mitochondrial membrane potential, the state of mitochondrial permeability transition pore (mPTP), the rate of AM apoptosis, the expression of Bcl-2 and Caspase-3 proteins were detected by flow cytometry, microplate reader and Western blotting.@*RESULTS@#Compared with the control group, in the I/R group, the ratio of red to green fluorescence and the absorbance decreased significantly, the percentage of apoptotic cells increased obviously, the apoptotic rate was 43.81%±2.40%, Caspase-3 protein expression increased, Bcl-2 protein expression decreased. While compared with the I/R group, in the I/R+SO2 group, the ratio of red to green fluorescence and the absorbance increased significantly; the apoptotic rate decreased to 37.01%±1.93%, Caspase-3 protein expression decreased, Bcl-2 protein expression increased.@*CONCLUSION@#Exogenous SO2 has the effect of accelerating AM apoptosis by stimulating mPTP to open and mitochondrial membrane potential to decrease; besides, exogenous SO2 could stimulate AM to secrete more anti-inflammatory cytokines and less inflammatory cytokines. In conclusion, exogenous SO2 can reduce macrophage apoptosis by inhibiting mitochondrial pathways.
Subject(s)Acute Lung Injury , Animals , Apoptosis , Ischemia , Macrophages, Alveolar , Male , Rats , Rats, Sprague-Dawley , Reperfusion Injury , Sulfur Dioxide
ABSTRACT Idiopathic pulmonary hemosiderosis is a potentially fatal disease that results from episodes of alveolar hemorrhage of unknown origin. The clinical spectrum is varied, and anemia may constitute the only manifestation of illness, preceding other signs and symptoms by several months. We present the case of a 4 year-old child presenting with fever, vomiting and prostration, associated with pallor. He had microcytic and hypochromic anemia refractory to iron therapy. Gastrointestinal bleeding was ruled out after negative extensive etiological investigation. Subsequently, pulmonary infiltrates suggestive of alveolar hemorrhage were observed in the chest radiography. The cytological exam of the bronchoalveolar lavage showed hemosiderin-laden macrophages. After the etiological study, the diagnosis of idiopathic pulmonary hemosiderosis was made by exclusion. He was initiated on corticosteroid therapy, later associated to an immunosuppressive agent, with subsequent correction of anemia and of the radiological pattern. The patient is currently asymptomatic.
RESUMO A hemossiderose pulmonar idiopática é uma doença potencialmente fatal que cursa com episódios de hemorragia alveolar de etiologia desconhecida. As manifestações clínicas são variadas, e a anemia pode constituir o único sinal de doença, precedendo em vários meses os outros sinais e sintomas. Apresenta-se o caso de criança de 4 anos, com febre, vômitos e prostração, associados à palidez. Apresentava anemia microcítica e hipocrômica, refratária à terapêutica com ferro. A hipótese diagnóstica de sangramento gastrintestinal foi excluída, após investigação etiológica extensa, inconclusiva. Posteriormente, em radiografia torácica, foram observados infiltrados sugestivos de hemorragia alveolar. O exame citológico do lavado broncoalveolar mostrou macrófagos com depósitos de hemossiderina. Após estudo etiológico, assumiu-se, por exclusão, o diagnóstico de hemossiderose pulmonar idiopática. Foi iniciada terapêutica com corticoides, associada posteriormente a imunossupressor, com correção subsequente da anemia e do padrão radiológico, encontrando-se, atualmente, assintomático.
Subject(s)Humans , Male , Child, Preschool , Anemia, Iron-Deficiency/etiology , Hemorrhage/etiology , Hemosiderosis/complications , Lung Diseases/complications , Hemoglobins/analysis , Bronchoalveolar Lavage Fluid/cytology , Macrophages, Alveolar/cytology , Anemia, Iron-Deficiency/blood , Hemorrhage/diagnostic imaging , Hemosiderosis/blood , Lung Diseases/blood
The porcine reproductive and respiratory syndrome virus (PRRSV) is a globally ubiquitous swine viral pathogen that causes major economic losses worldwide. We previously reported an over-attenuated phenotype of cell-adapted PRRSV strain CA-2-P100 in vivo. In the present study, CA-2-P100 was serially propagated in cultured porcine alveolar macrophage (PAM) cells for up to 20 passages to obtain the derivative strain CA-2-MP120. Animal inoculation studies revealed that both CA-2-P100 and CA-2-MP120 had decreased virulence, eliciting weight gains, body temperatures, and histopathologic lesions similar to those in the negative control group. However, compared to CA-2-P100 infection, CA-2-MP120 yielded consistently higher viremia kinetics and enhanced antibody responses in pigs. All pigs inoculated with CA-2-MP120 developed viremia and seroconverted to PRRSV. During 20 passages in PAM cells, CA-2-MP120 acquired 15 amino acid changes that were mostly distributed in nsp2 and minor structural protein-coding regions. Among these changes, 6 mutations represented reversions to the sequences of the reference CA-2 and parental CA-2-P20 strains. These genetic drifts may be hypothetical molecular markers associated with PRRSV macrophage tropism and virulence. Our results indicate that the PAM-passaged CA-2-MP120 strain is a potential candidate for developing a live, attenuated PRRSV vaccine.
Subject(s)Animals , Antibody Formation , Body Temperature , Genetic Drift , Humans , Kinetics , Macrophages , Macrophages, Alveolar , Parents , Phenotype , Porcine Reproductive and Respiratory Syndrome , Porcine respiratory and reproductive syndrome virus , Swine , Tropism , Vaccines, Attenuated , Viremia , Virulence , Weight Gain
Evaluation of alveolar macrophage functions of cattle is an important tool in order to assess whether measures taken during the cattle husbandry can decrease the respiratory tract defense. The aim of this study was to determine whether dexamethasone used at therapeutic dose can affect the oxidative metabolism of alveolar macrophages of cattle. This was evaluated by two tests, the fluorometric and colorimetric. The similarity of the results was studied, using alveolar macrophages of six healthy cattle, obtained from bronchoalveolar lavage on a basal and an immunosuppressant moment after the application of dexamethasone. For the fluorometric test, alveolar macrophages were incubated with Staphylococcus aureus and 2'-7'dichlorohidroflurescein, and analyzed by flow cytometer. For the colorimetric test, alveolar macrophages were incubated with Phorbol 12- miristate-13 acetate and nitroblue tetrazolium, dissolved and analyzed in a spectrophotometer. It was noted that dexamethasone therapeutic dose (0.05 mg/kg) reduced the functions of alveolar macrophages from healthy bovine. This result was observed by both tests with the difference that the flow cytometry assay was more informative for identifying which specific cellular function has been compromised.(AU)
A avaliação das funções de macrófagos alveolares de bovinos é uma ferramenta importante no intuito de analisar se medidas adotadas durante a criação desses animais diminuem a defesa do trato respiratório, aumentando a susceptibilidade à pneumonia. O objetivo deste trabalho foi verificar se a dexametasona em dose terapêutica afeta o metabolismo oxidativo de macrófagos alveolares de bovinos por meio de duas provas, a fluorimétrica e a colorimétrica, avaliando a similaridade dos resultados. Para tanto, utilizaram-se macrófagos alveolares de seis bovinos sadios, obtidos por meio de lavados broncoalveolares em um momento basal e em um momento imunossupressor, após a aplicação da dexametasona. Na prova fluorimétrica, os macrófagos alveolares foram incubados com Staphylococcus aureus e o fluorófilo 2'-7' diclorohidrofluresceína e analisados por citometria de fluxo. Na prova colorimétrica, os macrófagos alveolares foram incubados com Phorbol 12- miristate-13 acetate e nitroazul de tetrazolium, dissolvidos e analisados em espectofotômetro. Observou-se que a dexametasona em dose terapêutica (0,05 mg/kg) reduziu as funções dos macrófagos alveolares de bovinos sadios, sendo este resultado obtido em ambas as provas, com a diferença de que o teste de citometria de fluxo foi mais esclarecedor por identificar qual função celular específica foi comprometida.(AU)
Subject(s)Animals , Cattle , Respiratory System , Macrophages, Alveolar , Pneumonia , Staphylococcus aureus , Dexamethasone
A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, CD3⁺CD4⁺, CD3⁺CD8⁺, CD3⁺CD4⁺CD8⁺ T-lymphocytes, and PRRSV-specific interferon (IFN)-γ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages (CD172α⁺PRRSV-N⁺ PAM) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of CD172α⁺PRRSV-N⁺ PAM (p 0.1) with infection parameters. The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.
Subject(s)Adaptive Immunity , Antibodies , Antibodies, Neutralizing , Interferons , Lung , Lymph Nodes , Lymphoid Tissue , Macrophages, Alveolar , Phenotype , Plasma , Porcine Reproductive and Respiratory Syndrome , Porcine respiratory and reproductive syndrome virus , T-Lymphocytes , Viral Load
The inhalation of toxic environmental particles is a worldwide public health issue. To avoid the pulmonary damage, the lungs contain the alveolar macrophages, which are the primary defense of the innate immune system, since it engulfs the toxic or allergic particles. Morphologically, particulate matter inside of macrophage is observed as numerous round dark granules of various size. In guinea pig, the inhalation of fine particles in real time showed single round dark granules inside of the macrophages. After particles exposure, the alveolar macrophage can activate some cytokines such as TNF-α, IL-1β, IL-6, IL-8, and GM-CSF, which increases the inflammatory response or to activate the Th2 response. The alveolar macrophage interacts with bronchial and bronchiolar epithelium, heart, and blood vessels producing a variety of problems, such as nonfatal heart attacks, irregular heartbeat, decreased lung function, and increases respiratory symptoms such as irritation of the airways, coughing or difficulty breathing, aggravated asthma, and produce premature death in people with heart or lung disease.
La inhalación de partículas tóxicas ambientales es un problema de salud pública en todo el mundo. Para prevenir el daño, los pulmones contienen a los macrófagos alveolares, los cuales son la defensa primaria del sistema inmune, ya que fagocitan los tóxicos o partículas alérgicas. Morfológicamente, el material particulado dentro de los macrófagos alveolares se observa como numerosos gránulos redondos de varios tamaños. En cobayos, la inhalación de partículas finas en tiempo real mostró gránulos redondos oscuros dentro de los macrófagos. Después de la exposición a las partículas, el macrófago alveolar puede activar algunas citocinas como TNF-α, IL-1β, IL-6, IL-8, and GM-CSF, las cuales incrementan la respuesta inflamatoria o activan la respuesta Th2. El macrófago alveolar interactúa con el epitelio bronquial y bronquiolar, corazón y vasos sanguíneos, produciendo una variedad de problemas, tales como afecciones cardíacas, arritmias, disminución de la función pulmonar, e incrementa los síntomas respiratorios como irritación de las vías respiratorias, tos, dificultad para respirar, agrava el asma y produce muertes prematuras en personas con enfermedades cardiacas y pulmonares.
Subject(s)Animals , Guinea Pigs , Macrophages, Alveolar , Particulate Matter/adverse effects , Foreign-Body Reaction , Phagocytosis
The present study evaluated the impact that orchiectomy, a routine but painful intervention in bovine husbandry, can cause on pulmonary immunity. To identify whether orchiectomy can impair lung defense, analyses of serum cortisol concentration and of alveolar macrophage and their function (phagocytosis and respiratory burst) were evaluated. Sixteen Holstein bulls (7 mo old, 250±50kg of body weight BW) were divided in two homogeneous groups - the castrated group and the sham group - and the sample were collected on Days -7, 1 and 7 relative to the day of the procedure. Serum cortisol concentration statistically increased on Days 1 and 7 (D-7: 4,97±1,28ng/ml; D1: 6,35 ±1,10ng/ml; D7: 8,28±1,94ng/ml) after castration and these variables seem to impact the alveolar macrophage percentage on D1 (D-7: 76,86±3,44%; D1: 60,92±2,44%; D7: 74,17±2,56%) and their respective function of phagocytosis (P) and the oxidative burst (OB) on Days 1 and 7 for the castrated group (P D-7: 56,25±15,63 arbitrary values; D1: 54,75±14,07 arbitrary values; D7: 31,77±8,44 arbitrary values; and OB D-7: 222,34±39,52 arbitrary values; D1: 135,25±37,68 arbitrary values; D7: 117,73±18,17 arbitrary values). These results indicate that surgical castration affected lung defense until seven days after the practice, so the pulmonary cell function was impaired for a period higher than that reported in the literature.(AU)
O presente estudo avaliou o impacto que a orquiectomia, uma intervenção dolorosa comumente realizada durante a criação de bovinos, pode causar na imunidade pulmonar. Para tanto, foram realizadas dosagens de cortisol sérico, bem como a análise de macrófagos alveolares e suas funções (fagocitose e metabolismo oxidativo) de 16 bovinos da raça Holandesa preto e branco (sete meses de idade, 250±50kg de peso vivo). Esses animais foram divididos aleatoriamente em dois grupos homogêneos - grupo castrado e grupo controle - e foram avaliados nos dias -7, 1 e 7, relativos ao dia do procedimento cirúrgico, que foi realizado no dia 0. A concentração de cortisol sérico aumentou estatisticamente nos dias 1 e 7 em relação ao grupo controle (D-7: 4,97±1,28ng/mL; D1: 6,35 ±1,10ng/mL; D7: 8,28±1,94ng/mL). Notou-se diminuição de macrófagos alveolares no D1 (D-7: 76,86±3,44%; D1: 60,92±2,44%; D7: 74,17±2,56%) e de suas funções de fagocitose (F) e metabolismo oxidativo (MO) nos dias 1 e 7 (F D-7: 56,25±15,63 valores arbitrários; D1: 54,75±14,07 valores arbitrários; D7: 31,77±8,44 valores arbitrários; e MO D-7: 222,34±39,52 valores arbitrários; D1: 135,25±37,68 valores arbitrários, D7: 117,73±18,17 valores arbitrários) para o grupo castrado. Esses resultados demonstram que a orquiectomia afeta as defesas pulmonares por até sete dias após a prática, período superior ao relatado pela literatura.(AU)
Subject(s)Animals , Male , Cattle , Immunity, Innate , Orchiectomy/adverse effects , Orchiectomy/veterinary , Cell Respiration/physiology , Hydrocortisone/blood , Macrophages, Alveolar/physiology , Phagocytosis
We investigated whether Nd2O3 treatment results in cytotoxicity and other underlying effects in rat NR8383 alveolar macrophages. Cell viability assessed by the MTT assay revealed that Nd2O3 was toxic in a dose-dependent manner, but not in a time-dependent manner. An ELISA analysis indicated that exposure to Nd2O3 caused cell damage and enhanced synthesis and release of inflammatory chemokines. A Western blot analysis showed that protein expression levels of caspase-3, nuclear factor-κB (NF-κB) and its inhibitor IκB increased significantly in response to Nd2O3 treatment. Both NF-κB and caspase-3 signaling were activated, suggesting that both pathways are involved in Nd2O3 cytotoxicity.
Subject(s)Animals , Caspase 3 , Metabolism , Cell Line , Macrophages, Alveolar , NF-kappa B , Metabolism , Neodymium , Toxicity , Oxides , Toxicity , Rats , Toxicity Tests
BACKGROUND/OBSECTIVE: Airway inflammation by eosinophils, neutrophils and alveolar macrophages is a characteristic feature of asthma that leads to pathological subepithelial thickening and remodeling. Our previous study showed that oxidative stress in airways resulted in eosinophilia and epithelial apoptosis. The current study investigated whether glutathione-containing dry yeast extract (dry-YE) ameliorated eosinophilia, goblet cell hyperplasia and mucus overproduction. MATERIALS/METHOD: This study employed 2 µg/mL lipopolysaccharide (LPS)- or 20 ng/mL eotaxin-1-exposed human bronchial epithelial cells and ovalbumin (OVA)-challenged mice. Dry-YE employed in this study contained a significant amount of glutathione (140 mg in 100 g dry yeast). RESULTS: Human bronchial epithelial cell eotaxin-1 and mucin 5AC (MUC5AC) were markedly induced by the endotoxin LPS, which was dose-dependently attenuated by nontoxic dry-YE at 10-50 µg/mL. Moreover, dry-YE inhibited the MUC5AC induction enhanced by eotaxin-1, indicating that eotaxin-1-mediated eosinophilia may prompt the MUC5AC induction. Oral supplementation with 10-100 mg/kg dry-YE inhibited inflammatory cell accumulation in airway subepithelial regions with a reduction of lung tissue level of intracellular adhesion molecule-1. In addition, ≥ 50 mg/kg dry-YE diminished the lung tissue levels of eotaxin-1, eosinophil major basic protein and MUC5AC in OVA-exposed mice. Alcian blue/periodic acid schiff staining revealed that the dry-YE supplementation inhibited goblet cell hyperplasia and mucus overproduction in the trachea and bronchiolar airways of OVA-challenged mice. CONCLUSIONS: Oxidative stress may be involved in the induction of eotaxin-1 and MUC5AC by endotoxin episode and OVA challenge. Dry-YE effectively ameliorated oxidative stress-responsive epithelial eosinophilia and mucus-secreting goblet cell hyperplasia in cellular and murine models of asthma.
Subject(s)Animals , Apoptosis , Asthma , Chemokine CCL11 , Eosinophil Major Basic Protein , Eosinophilia , Eosinophils , Epithelial Cells , Glutathione , Goblet Cells , Humans , Hyperplasia , Inflammation , Lung , Macrophages, Alveolar , Mice , Mucin 5AC , Mucins , Mucus , Neutrophils , Ovalbumin , Ovum , Oxidative Stress , Trachea , Yeasts
Porcine alveolar macrophages (PAMs) represent the first line of defense in the porcine lung after infection with porcine circovirus type 2 (PCV2) via the respiratory tract. However, PCV2 infection impairs the microbicidal capability of PAMs and alters cytokine production and/or secretion. At present, the reason for the imbalance of cytokines has not been fully elucidated, and the regulatory mechanisms involved are unclear. In this study, we investigated the expression levels and regulation of interleukin-1beta (IL-1β) and IL-10 in PAMs following incubation with PCV2 in vitro. Levels of IL-1β and IL-10 increased in PAM supernatants, and the distribution of nuclear factor kappa B (NF-κB) p65 staining in nucleus, expression of MyD88 and p-IκB in cytoplasm, and DNA-binding activity of NF-κB increased after incubation with PCV2, while p65 expression in PAM cytoplasm decreased. However, when PAMs were co-incubated with PCV2 and small interfering RNA targeting MyD88, those effects were reversed. Additionally, mRNA expression levels of Toll-like receptors (TLR)-2, -3, -4, -7, -8, and -9 increased when PAMs were incubated with PCV2. These results show that PCV2 induces increased IL-1β and IL-10 production in PAMs, and these changes in expression are related to the TLR–MyD88–NF-κB signaling pathway.
Subject(s)Circovirus , Cytokines , Cytoplasm , In Vitro Techniques , Interleukin-10 , Interleukin-1beta , Lung , Macrophages, Alveolar , NF-kappa B , Respiratory System , RNA, Messenger , RNA, Small Interfering , Toll-Like Receptors
La neumoconiosis es caracterizada por el depósito nodular difuso de polvo en los pulmones como resultado de la exposición prolongada a polvo bituminoso o de antracita en los trabajadores de las minas de carbón. La neumoconiosis de los mineros del carbón también se denomina enfermedad del pulmón negro o antracosis. Un minero del carbón que padece o desarrolla una Antracosis puede presentar numerosos nódulos redondeados pulmonares en poco tiempo. Dichos nódulos aparecen en ocasiones en ausencia de una antracosis simple. A nivel histológico pueden parecerse a los nódulos reumatoides, pero tienen una zona periférica de inflamación aguda. Estos nódulos representan la respuesta inmunológica a la diátesis reumatoide asociada. En Colombia la minería es un factor estratégico a nivel económico para su desarrollo. Sin embargo, existen factores negativos derivados de ésta que giran en una carrera sin control ni reglas claras; esta actividad hace trámite en el territorio, arrastrando una estela de problemas sobre la sociedad, el ambiente, el bienestar y la salud de las personas.
Pneumoconiosis is characterized by diffuse nodular dust in the lungs as a result of prolonged exposure of workers in coal mines to bituminous dust or anthracite. Pneumoconiosis presented in coal miners is also called black lung disease lung. A coal miner who develops Anthracosis can present numerous pulmonary rounded nodules in a short time. These nodules appear sometimes in the absence of a simple anthracosis. Histologically they may look like resemble rheumatoid nodules, with the difference; they have a peripheral area of acute inflammation. These nodes represent the immune response associated to rheumatoid diathesis. In Colombia, mining is a strategy for economical development. However, there are negative factors arising rotating it in a race without control or clear rules; This activity is pending in the territory, dragging a trail of problems on society, the environment, welfare and People's health
A pneumoconiose é caracterizada por poeira nodular difusa nos pulmões como resultado da exposição prolongada a poeiras betuminosas ou antracite nas minas de carvão. Pneumoconiose de mineiros de carvão também chamado preto ou preto doença pulmonar pulmão. Um mineiro de carvão que sofre ou desenvolve uma antracose pode apresentar numerosos nódulos pulmonares arredondados em um curto espaço de tempo. Estes nódulos aparecem às vezes na ausência de uma antracose simples. Histologicamente podem assemelhar-se a nódulos reumatóides, mas têm uma área periférica de inflamação aguda. Esses nódulos representam a resposta imune associada à diátese reumatóide. A mineração na Colômbia é um fator estratégico economicamente para o desenvolvimento. No entanto, existem fatores negativos decorrent surgindo girando-o em uma corrida sem controle ou regras claras. Esta atividade está pendente no território, arrastando um rastro de problemas na sociedade, o meio ambiente, bem-estar e da saúde das pessoas.
Subject(s)Humans , Male , Adult , Pneumoconiosis , Macrophages, Alveolar , Colombia , Dyspnea , Anthracosis
A citologia é um importante exame complementar utilizado para a detecção de alterações no perfil celular do trato respiratório e auxílio no diagnóstico de doenças. No entanto, como os primeiros meses de vida compõem o período de adaptação à vida extrauterina resultando em possíveis alterações das populações celulares, é essencial a padronização das características comuns aos animais sadios, possibilitando a identificação e análise de qualquer mudança no quadro esperado. O objetivo desta pesquisa foi caracterizar o perfil celular por citologia do lavado bronco alveolar obtido semanalmente por broncoscopia de dez bezerros durante os três primeiros meses de vida. A análise estatística foi realizada utilizando-se o software Minitab® 15.0, empregando-se o teste T pareado para as amostras paramétricas e Mann-Whitney para as amostras não paramétricas, considerando nivel de significância P≤0,05. Os resultados obtidos apontaram que o perfil celular do lavado bronco alveolar de bezerros hígidos durante os primeiros 90 dias de vida apresentou predomínio de macrófagos, com média geral de 63,17%, seguido de 33,69% de neutrófilos. Porém observou-se diminuição significativa na porcentagem de macrófagos (P=0,04) e aumento de neutrófilos (P=0,05) ao longo dos momentos, comprovada pela forte correlação negativa entre as porcentagens de macrófagos e neutrófilos ao longo dos momentos. Não houve diferença entre as porcentagens de células gigantes, células epiteliais, linfócitos, eosinófilos e basófilos durante o experimento. Apesar dos resultados serem influenciados por fatores ambientais e de manejo, os resultados dessa pesquisa fornecem subsídios para a identificação de alterações críticas que descaracterizem o perfil celular de bezerros acometidos por doenças respiratórias.
Cytology is an important supplementary test used for the detection of changes in the mobile profile of the respiratory tract and aid in the diagnosis of diseases. However, as the first few months of life make up the period of adjustment to extrauterine life resulting in possible changes in cellular populations, it is essential to the standardization of the characteristics common to healthy animals, enabling the identification and analysis of any change in the expected frame. The objective of this research was to characterize the cellular profile by cytology of bronchoalveolar lavage obtained weekly for bronchoscopy of ten calves during the first three months of life. Statistical analysis was performed using the Minitab® 15.0 software, using the paired T test for parametric samples, and Mann-Whitney for nonparametric samples, considering significance level as P≤0,05. The results obtained showed that the profile of bronchoalveolar lavage of healthy calves during the first 90 days of life showed a predominance of macrophages, with medium average of 63.17%, followed by 33.69% neutrophils. However it was observed significant decrease in percentage of macrophages (P=0.04) and increased neutrophils (P=0,05) over the times, proven by the strong negative correlation between percentages of macrophages and neutrophils along the times. There was no difference between the percentages of giant cells, epithelial cells, lymphocytes, eosinophils and basophils during the experiment. Although the results are influenced by environmental factors and management, the results of this research provide subsidies for the identification of critical changes in cell profile of calves affected by respiratory diseases.
Subject(s)Animals , Cattle , Macrophages, Alveolar , Neutrophils , Lung/cytology , Reference Standards , Bronchoscopy/veterinary
In the mammalian lung, respiratory macrophages provide front line defense against invading pathogens and particulate matter. In birds, respiratory macrophages are known as free avian respiratory macrophages (FARM) and a dearth of the cells in the avian lung has been purported to foreordain a weak first line of pulmonary defense, a condition associated with high mortality of domestic birds occasioned by respiratory inflictions. Avian pulmonary mechanisms including a three tiered aerodynamic filtration system, tight epithelial junctions and an efficient mucociliary escalator system have been known to supplement FARM protective roles. Current studies, however, report FARM to exhibit an exceptionally efficient phagocytic capacity and are effective in elimination of invading pathogens. In this review, we also report on effects of selective synthetic peroxisome proliferator activated receptor gamma (PPAR Î³) agonists on non phlogistic phagocytic properties in the FARM. To develop effective therapeutic interventions targeting FARM in treatment and management of respiratory disease conditions in the poultry, further studies are required to fully understand the role of FARM in innate and adaptive immune responses.
Subject(s)Animals , Birds/immunology , Macrophages, Alveolar/physiology , Lung/immunology , Particle Size , Phagocytes/immunology , Phagocytosis , Respiratory Tract Infections/immunology , Respiratory Tract Infections/veterinary , PPAR gamma/physiology , Lung/cytology
Mycobacterium tuberculosis (Mtb) causing tuberculosis as an intracellular pathogen initially infects alveolar macrophages following aerosol inhalation. Thus, macrophages play a critical role in the establishment of Mtb infection and macrophage cell death, a common outcome during Mtb infection, may initiate host- or pathogen-favored immune responses, resulting in facilitating protection or pathogenesis, respectively. In addition, virulent Mtb strains are known to inhibit apoptosis and consequently down-regulates immune response using a variety of strategies. In many recent studies have shown that virulent Mtb can either augment or reduce apoptosis by regulating expression of pro-apoptotic and anti-apoptotic proteins belonging to Bcl-2 family proteins. In this review, we will discuss and dissect the apoptotic pathways of Bcl-2 family proteins in Mtb-infected macrophages.
Subject(s)Apoptosis , Apoptosis Regulatory Proteins , Cell Death , Humans , Inhalation , Macrophages , Macrophages, Alveolar , Mycobacterium tuberculosis , Mycobacterium , Tuberculosis
In order to find potential therapeutic agents on lung inflammatory conditions, the extracts of Acanthopanax divaricatus var. albeofructus were prepared and its constituents were isolated. They include lignans such as (+)-syringaresinol (1), acanthoside B (2), salvadoraside (3) and acanthoside D (4), lariciresinol-9-O-beta-D-glucopyranoside (5) and phenylpropanoids such as 4-[(1E)-3-methoxy-1-propenyl]phenol (6), coniferin (7), and methyl caffeate (8). The extracts and several constituents such as compound 1, 6 and 8 inhibited the production of inflammatory markers, IL-6 and nitric oxide, from IL-1beta-treated lung epithelial cells and lipopolysaccharide (LPS)-treated alveolar macrophages. Furthermore, the extracts and compound 4 significantly inhibited lung inflammation in lipolysaccharide-treated acute lung injury in mice by oral administration. Thus it is suggested that A. divaricatus var. albeofructus and its several constituents may be effective against lung inflammation.
Subject(s)Eleutherococcus , Acute Lung Injury , Administration, Oral , Animals , Epithelial Cells , Interleukin-6 , Lignans , Lung , Macrophages , Macrophages, Alveolar , Mice , Nitric Oxide , Pneumonia
Ambroxol is used in COPD and asthma to increase mucociliary clearance and regulate surfactant levels, perhaps through anti-oxidant and anti-inflammatory activities. To determine the role and effect of ambroxol in an experimental model of asthma, BALB/c mice were sensitized to ovalbumin (OVA) followed by 3 days of challenge. Airway hyperresponsiveness (AHR), lung cell composition and histology, and cytokine and protein carbonyl levels in bronchoalveolar lavage (BAL) fluid were determined. Ambroxol was administered either before the first OVA challenge or was begun after the last allergen challenge. Cytokine production levels from lung mononuclear cells (Lung MNCs) or alveolar macrophages (AM) were also determined. Administration of ambroxol prior to challenge suppressed AHR, airway eosinophilia, goblet cell metaplasia, and reduced inflammation in subepithelial regions. When given after challenge, AHR was suppressed but without effects on eosinophil numbers. Levels of IL-5 and IL-13 in BAL fluid were decreased when the drug was given prior to challenge; when given after challenge, increased levels of IL-10 and IL-12 were detected. Decreased levels of protein carbonyls were detected in BAL fluid following ambroxol treatment after challenge. In vitro, ambroxol increased levels of IL-10, IFN-γ, and IL-12 from Lung MNCs and AM, whereas IL-4, IL-5, and IL-13 production was not altered. Taken together, ambroxol was effective in preventing AHR and airway inflammation through upregulation of Th1 cytokines and protection from oxidative stress in the airways.