1.Mac-1-mediated Uptake and Killing of Bordetella bronchiseptica by Porcine Alveolar Macrophages.
Jong Keuk LEE ; Lawrence B SCHOOK ; Mark S RUTHERFORD
Journal of Veterinary Science 2003;4(1):41-49
The role of Mac-1 as a receptor for Bordetella bronchiseptica infection of alveolar macrophages (AMphi) was examined using 6 strains (2 ATCC and 4 pathogenic field isolates) to assess B. bronchiseptica binding, uptake and replication in primary porcine AMphi. All B. bronchiseptica strains were rapidly killed by porcine serum in a dose- and time-dependent manner. However, heat-inactivated porcine serum (HIS) did not demonstrate any bacterial-killing activity, suggesting that complement may have a direct killing activity. All field isolates were more resistant to direct complement-mediated B. bronchiseptica killing. The uptake of B. bronchiseptica into AMphi was inhibited approximately 50% by antiMac-1 monoclonal antibodies in the medium. However, B. bronchiseptica phagocytosed in the presence of serum or HIS was not altered by anti-Mac-1 antibodies although more bacteria were internalized by addition of serum or HIS. These data suggest that Mac-1 is a target for direct uptake of B. bronchiseptica via opsoninindependent binding. The phagocytosed B. bronchiseptica, either via direct or serum-mediated binding, were efficiently killed by AMphi within 10 hr postinfection. This demonstrates that Mac-1-mediated B. bronchiseptica uptake is a bacterial killing pathway not leading to productive infections in AMphi.
Animals
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Antibodies, Bacterial/blood/immunology
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Bordetella bronchiseptica/*immunology
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Macrophage-1 Antigen/*immunology
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Macrophages, Alveolar/*immunology
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Phagocytosis
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Protein Binding
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Swine/*immunology/*microbiology
2.Effect of human silicotic alveolar macrophages on the expression of the collagen type I in human embryonic lung fibroblasts.
Xiao-hui HAO ; Xian-hua WANG ; Li ZHANG ; Fang YANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2007;25(2):69-72
OBJECTIVETo study the effect of the cultured supernatant of human silicotic alveolar macrophages (AM) on the expression of the collagen type I in human embryonic lung fibroblasts.
METHODSHuman alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 18 h. Then the cultured supernatant were used to culture human embryonic lung fibroblasts for 6 h, 12 h, 18 h, 24 h, 36 h, 48 h, 72 h. Then detected collagen anabolism and secretion with (3)H-proline detected the expression of the procollagen type I in the fibroblast with immunological method detected the quantity of collagen Type I in FB supernatant with Western blot.
RESULTSThe anabolism and secretion of collagen were increased in cultured supernatant of silicotic AM exposed to SiO(2), Along with the time, the expression of collagen type I increased. In cultured supernatant of silicotic AM exposed to SiO(2), ((3)H-proline: 1096.500 +/- 76.400, 707.000 +/- 62.160, OD: 0.314 +/- 0.011, OD: 14.218 +/- 0.342.
CONCLUSIONSiO(2) may affect the expression of collagen through AM mediation and participate in the formation of lung fibrosis.
Adult ; Cells, Cultured ; Collagen Type I ; metabolism ; Fibroblasts ; metabolism ; Humans ; Lung ; metabolism ; Macrophages, Alveolar ; immunology ; Male ; Silicosis ; immunology
3.Alveolar macrophages modulate allergic inflammation in a murine model of asthma.
Bo Ram BANG ; Eunyoung CHUN ; Eun Jin SHIM ; Hyun Seung LEE ; Soo Yeon LEE ; Sang Heon CHO ; Kyung Up MIN ; You Young KIM ; Heung Woo PARK
Experimental & Molecular Medicine 2011;43(5):275-280
The role of alveolar macrophages (AMs) in the pathogenesis of asthma is still unknown. The aim of the present study was to investigate the effects of AM in the murine model of asthma. AMs were selectively depleted by liposomes containing clodronate just before allergen challenges, and changes in inflammatory cells and cytokine concentrations in bronchoalveolar lavage (BAL) fluid were measured. AMs were then adoptively transferred to AM-depleted sensitized mice and changes were measured. Phenotypic changes in AMs were evaluated after in vitro allergen stimulation. AM-depletion after sensitization significantly increased the number of eosinophils and lymphocytes and the concentrations of IL-4, IL-5 and GM-CSF in BAL fluid. These changes were significantly ameliorated only by adoptive transfer of unsensitized AMs, not by sensitized AMs. In addition, in vitro allergen stimulation of AMs resulted in their gaining the ability to produce inflammatory cytokines, such as IL-1beta, IL-6 and TNF-alpha, and losing the ability to suppress GM-CSF concentrations in BAL fluid. These findings suggested that AMs worked probably through GM-CSF-dependent mechanisms, although further confirmatory experiments are needed. Our results indicate that the role of AMs in the context of airway inflammation should be re-examined.
Animals
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Asthma/*immunology
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Bronchoalveolar Lavage Fluid/chemistry/cytology/immunology
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Cytokines/biosynthesis/immunology
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Disease Models, Animal
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Female
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Immunization
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Immunomodulation/*immunology
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Inflammation/*immunology
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Leukocytes/immunology
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Macrophages, Alveolar/*immunology
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Mice
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Mice, Inbred C57BL
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Ovalbumin/immunology
4.Effects of huai qi huang on cytokines Th1, Th2 and Th17 and phagocytosis of alveolar macrophages in rats with asthma.
Hong-Mei LI ; Qing-Nan HE ; Xiao-Yan LI ; Lan-Jun SHUAI ; Pin ZHOU ; Zhu-Wen YI
Chinese Journal of Contemporary Pediatrics 2011;13(9):747-750
OBJECTIVETo study the effects of huai qi huang, a traditional Chinese medicine, on cytokines Th1, Th2 and Th17 levels and alveolar macrophage phagocytosis in asthmatic rats sensitized by ovalbumin (OVA).
METHODSForty male Sprague-Dawley rats were randomly divided into five groups: normal control, untreated asthma, budesonide-treated, huai qi huang-treated and budesonide+huai qi huang-treated asthma (n=8 each). Asthma was induced by OVA sensitization and challenge. The levels of IL-4, IFN-γ and IL-17 in plasma and bronchoalveolar lavage fluid (BALF) were measured using ELISA. The phagocytosis of alveolar macrophages which were isolated and purified from BALF was evaluated by the colorimetric assay.
RESULTSThe levels of IL-4 and IL-17 increased, in contrast, the IFN-γ level decreased in plasma and BALF in the untreated asthma group compared with those in the normal control group. The IFN-γ level in the huai qi huang-treated asthma group was higher than that in the untreated asthma group. The IFN-γ level increased and the IL-17 level decreased more significantly in the budesonide+huai qi huang-treated asthma group when compared with the budesonide and huai qi huang alone treatment groups. The phagocytosis of alveolar macrophages in the untreated asthma group was lower than that in the normal control group. Huai qi huang alone or combined with budesonide increased the phagocytosis of alveolar macrophages compared with the normal control, untreated asthma and budesonid-treated asthma groups. The levels of IFN-γ in plasma and BALF were positively correlated with the phagocytosis of alveolar macrophages.
CONCLUSIONSThe levels of IL-4 and IL-17 increase and the IFN-γ level decreases in plasma and BALF, and the phagocytosis of alveolar macrophages decreases in asthmatic rats. Huai qi huang treatment may increase the IFN-γ expression in plasma and BALF and the phagocytosis of alveolar macrophages in asthmatic rats. There is a synergistic effect between huai qi huang and glucocorticoids.
Animals ; Asthma ; drug therapy ; immunology ; Cytokines ; biosynthesis ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; Medicine, Chinese Traditional ; Phagocytosis ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes, Helper-Inducer ; immunology ; Th1 Cells ; immunology ; Th17 Cells ; immunology ; Th2 Cells ; immunology
5.Effect of ginsenoside Rh2 on immunocompetence of alveolar macrophages in patients with non-small cell lung cancer.
Dong-bo ZHOU ; Cheng-ping HU ; Shuo LIANG ; Hong-zhong YANG
Journal of Central South University(Medical Sciences) 2007;32(5):868-872
OBJECTIVE:
To explore the effect of ginsenoside Rh2 (G-Rh2) on the excretion of cytotoxin-effecting molecule of alveolar macrophages (AM) in patients with non-small cell lung cancer (NSCLC).
METHODS:
The concentration of tumor necrosis factor (TNF-alpha) and NO in the bronchoalveolar lavage fluid (BALF) and the cultured supernatants of AM in 35 patients with NSCLC were measured by ELISA and enzyme method,and levels of TNF-alpha and NO in the cultured supernatants of AM after being cultivated with IFN-alpha, G-Rh2, and IFN-alpha+G-Rh2 were measured by the same method.
RESULTS:
AM in all the non-small cell lung cancer patients produced TNF-alpha and NO. The activity of TNF-alpha and NO was lower in the BALF and in the cultured supernatants of AM of the tumor-bearing lungs than that of the non-tumor-bearing lungs. The concentrations of TNF-alpha and NO in the cultured supernatants of AM cultivated with G-Rh2 were higher than those in the control (P<0.05), but there were no significant differences between the G-Rh2 group and IFN-alpha group (P>0.05). The concentrations of TNF-alpha and NO in the cultured supernatants of AM cultivated with both G-Rh2 and IFNalpha were obviously higher than those stimulated with IFNalpha or G-Rh2 (P<0.01) alone.
CONCLUSION
G-Rh2 can enhance the excretion of cytotoxin-effecting molecules of AM in patients with NSCLC. The changes are more distinctive when G-Rh2 and IFNalpha have coordinated action.
Adult
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Aged
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Bronchoalveolar Lavage Fluid
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chemistry
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Carcinoma, Non-Small-Cell Lung
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immunology
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Female
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Ginsenosides
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pharmacology
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Humans
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Lung Neoplasms
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immunology
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Macrophages, Alveolar
;
drug effects
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immunology
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metabolism
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Male
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Middle Aged
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Nitric Oxide
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metabolism
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Tumor Cells, Cultured
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Tumor Necrosis Factor-alpha
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metabolism
6.Immunohistochemical investigation of lungs with severe acute respiratory syndrome.
Zhao-hui LU ; Jie CHEN ; Yu-feng LUO ; Jin-ling CAO ; Jian-wei WAN ; De-tian WANG ; Hong-tu ZHANG ; Yong-qiang XIE
Acta Academiae Medicinae Sinicae 2003;25(5):508-511
OBJECTIVETo investigate the roles of different cells in the pulmonary lesions in the severe acute respiratory syndrome (SARS) patients.
METHODSThe monoclonal antibodies of CD8, CD20, CD34, LCA, CD56, CD68, and AE1/AE3 are used to demonstrate the different cells in the lung specimens of SARS patients in order to study the patterns of cell responses in this new disease. Meanwhile the HE stained slides were also carefully studied to compare with the results of immunohistochemical staining.
RESULTSThe number of capillaries increased and the capillaries clearly outlined the contour of alveolar wall from beginning to early stage of organization, the number of lymphocytes decreased sharply while the number of macrophage remarkably increased, together with proliferation of type II pneumocytes. The numbers of blood vessels decreased in the fibrotic and consolidated lung tissue, and the vessel cavities enlarged, losing the normal contour of alveolar septa.
CONCLUSIONSThe lesions in the lung from SARS patients are consisted of the tissue reaction to the inflammatory injury, including extensive exudation, capillary proliferation, fibrosis, and obvious infiltration of macrophages which may play a key role in the pathogenesis of pulmonary lesions of SARS.
Adult ; Antigens, CD ; immunology ; Antigens, CD20 ; immunology ; Antigens, CD34 ; immunology ; Antigens, Differentiation, Myelomonocytic ; immunology ; Capillaries ; pathology ; Edema ; pathology ; Female ; Fibrosis ; pathology ; Humans ; Immunohistochemistry ; Lung ; blood supply ; pathology ; Macrophages, Alveolar ; pathology ; Male ; Middle Aged ; Pulmonary Alveoli ; pathology ; Severe Acute Respiratory Syndrome ; pathology
7.Expression of nuclear factor-kappaB and its inhibitor in alveolar macrophages of patients with neonatal hyaline membrane disease.
Cui-qing LIU ; Lei CAO ; Hua-cheng ZHENG ; Xi-qun JIA ; Li-min KANG ; Lan-feng LI ; Su-zhe LIU
Chinese Journal of Pediatrics 2006;44(8):602-606
OBJECTIVEInflammatory reaction and injury in immature lungs are associated with activation of nuclear factor-kappa B (NF-kappaB) to trigger proinflammatory cytokine release, but the mechanism thereof is not fully understood. The present study was conducted to understand possible relationship between expression of NF-kappaB and its inhibitor and severity and outcome of neonates with hyaline membrane disease (HMD).
METHODSSerial samples of bronchoalveolar lavage fluid (BALF) were obtained during mechanical ventilation from 31 preterm infants with HMD. These infants were divided into two groups: survivors group [n = 22, birth weight (1500 +/- 320) g and gestational age (31.2 +/- 1.8) weeks] and nonsurvivors group [birth weight (1340 +/- 280) g, gestational age (30.8 +/- 2.1) weeks]. Nineteen preterm infants [birth weight (1470 +/- 280) g, gestational age (30.6 +/- 1.9) weeks] without respiratory disorders were enrolled as control subjects. Alveolar macrophages (AM) were isolated by differential adherence. AM was cultured and treated with lipopolysaccharide (LPS) for 1 hr. Then, nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for NF-kappaB expression. NF-kappaB inhibitor (IkappaB-alpha protein) in cytoplasmic extracts was detected by using Western blotting and IL-1beta and IL-8 in BALF by enzyme-linked immunosorbent assay (ELISA).
RESULTSNF-kappaB complexes were observed by EMSA, they were characterized by competition with cold oligonucleotide and p65-specific antibodies. The addition of an excess of cold oligonucleotide, corresponding to the NF-kappaB binding site, turned off the signal of the band, showing that the band was specific. An excess of an irrelevant oligonucleotide (corresponding to the SP-1) did not show any effect. The addition of an anti-p65 antibody caused the supershift of the two upper bands. After EMSA, the NF-kappaB complexes were quantified by using a ImageQuant software. NF-kappaB expression in AM at 24 hrs was higher in all the patients with HMD as compared with control subjects (survives/control, 34.1 vs 11.4 RDU, P < 0.01; nonsurvivors/control, 55.2 vs 11.4 RDU, P < 0.01). The NF-kappaB expression in AM at 72 hrs was higher than that in control subjects but not for nonsurvivors (survivors/control, 47.8 vs 25.6 RDU, P < 0.01; nonsurvivors/control, 21.8 vs 25.6, P > 0.05). The NF-kappaB expression in AM from nonsurvivors was depressed at 72 hrs as compared to 24 hrs (21.8 vs 55.2, P < 0.01), whereas the NF-kappaB expression in AM from survivors was still higher at 72 hrs than that at 24 hrs (47.8 vs 34.1, t = 4.43, P < 0.01).
CONCLUSIONAltered NF-kappaB activation in AM of BALF of neonates with HMD was observed, and it may be mediated by decreased IkappaB synthesis, increased IkappaB degradation, or both. In HMD nonsurvivors NF-kappaB translocation was hampered upon LPS activation.
Birth Weight ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Culture Techniques ; Cell Nucleus ; drug effects ; metabolism ; Cytoplasm ; drug effects ; metabolism ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Female ; Gestational Age ; Humans ; Hyaline Membrane Disease ; immunology ; therapy ; I-kappa B Proteins ; immunology ; Infant, Newborn ; Infant, Premature ; immunology ; Interleukin-1beta ; immunology ; Interleukin-8 ; immunology ; Lipopolysaccharides ; pharmacology ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; NF-KappaB Inhibitor alpha ; NF-kappa B ; immunology ; Respiration, Artificial ; Severity of Illness Index ; Time Factors
8.Role of Fas/FasL pathway-mediated alveolar macrophages releasing inflammatory cytokines in human silicosis.
San Qiao YAO ; Qin Cheng HE ; Ju Xiang YUAN ; Jie CHEN ; Gang CHEN ; Yao LU ; Yu Ping BAI ; Chun Min ZHANG ; Yang YUAN ; Ying Jun XU
Biomedical and Environmental Sciences 2013;26(11):930-933
Adult
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Antibodies, Monoclonal
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pharmacology
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Bronchoalveolar Lavage Fluid
;
cytology
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Cells, Cultured
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Cytokines
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biosynthesis
;
blood
;
secretion
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Fas Ligand Protein
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antagonists & inhibitors
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metabolism
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Humans
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Macrophages, Alveolar
;
immunology
;
metabolism
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Middle Aged
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Occupational Exposure
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analysis
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Signal Transduction
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Silicon Dioxide
;
adverse effects
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Silicosis
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blood
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immunology
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fas Receptor
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antagonists & inhibitors
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metabolism