Objective To investigate the feasibility of dendritic cells ( DCs ) as vector of immunotherapy through intratracheal injection. Methods The DCs obtained from the bone marrow of BALB/ c mice were cultured and isolated with CD11c-positive magnetic beads. Then DCs were overloaded with ovalbumin peptide 323-339 ( OVA 323-339) for 24 hours. The mice in the DC-OVA group were intratrachelly injected DCs overloaded with OVA 323-339 in dose of 2 ×106 cells per mouse. The mice in thenegative control group were intratracheally injected with DCs untreated by OVA 323-339. On the second day,all mice were challenged with 1% OVA in PBS lasting for five days. The asthma animal model established by classic method was used for the positive control. Pathologic changes in lung and cell numbers in bronchoalveolar lavage fluid ( BALF) were assayed 24 hours after challenged. Results Just like the lung tissues from the mice asthma models, the lung tissues from the mice instilled with DCs overloaded with allergen OVA 323-339 showed extensive inflammatory cells infiltration, most of which were eosinophils, neutrophils and lymphocytes. The lung tissues in the DC group showed no obvious inflammation. There were more cells in BALF in the DC-OVA group than that in the DC group. OVA-specific IgE in serum from the DC-OVA group was not significantly different from that in the mice asthma models [ ( 48. 22 ±4. 76) U/mL vs. ( 52. 75 ±4. 03) U/mL, P gt;0. 05] . Conclusion DCs overloaded antigen has the ability of transferring of antigen effectively and may be used as vectors of immunotherapy.
0bjective To study the effect of bacterial infection on acute exacerbation of chronic obstructive pulmonary disease(AECOPD),and to compare the airway inflammation caused by different isolated bacteria.Methods A total of 159 sputum samples were collected from AECOPD patients diagnosed according to GOLD 2004 standard,in which conventional culture and identification of bacteria was conducted.The patients with purulent sputa were divided into different groups according to bacteria separated.Levels of IL-6.IL-8 and TNF-α in sputum supernatant were assayed and compared in different bacteria groups.the purulent sputum without isolated bacteria group(NG)and normal control group(NC). Results One hundred and twenty-nine strains of bacteria were isolated in 159 qualified sputa,including 26 strains of Klebsiella pneumoniae(KB),21 strains of Hemophilus influenza(Hi),17 strains of Pseudomonas aeruginosa(PA),37 strains of Haemophilus parainflb~enzae(HP)(mixed infection not included)and 28 strains of other bacteria.Among of all samples,20 were double infection of Haernophilus parainfluenzae with another bacterium.Ninety-seven purulent sputa were collected.According to bacteria isolated,these sputa were divided into five groups,named HP(24 samples),Hi(20 samples),PA(16 samples),KB(19 samples)and NG(18 samples).Contrast to NC,concentration of IL-8 and TNF-α rose in sputa from which PA,Hi,KB were isolated(Plt;0.05).The level increased much more in sputa from which PA and Hi were isolated compared with KB(Plt;0.05).Concentration of IL-6 rose in sputa of each group collected from AECOPD patients contrast to sputa collected from NC(Plt;0.05),without significant differences among all other group except for NC.Conclusions Bacterial infection plays an important role in AECOPD characterized with high level of inflammatory factors especially when PA,Hi,KB were infected bacteria.This study provides evidence for antibacterial therapy in AECOPD patients.
Objective To study the effect of nontypeable Hemophilus influenzae(NTHi) strain ATCC49247 on proinflammatory cytokines expression of human A549 lung epithelial cell line. Methods Confluent A549 cells were co-incubated with NTHi, NTHi+Erythromycin(10 mg/L), NTHi+Gentamicin(100 mg/L), and NTHi+Dexamethasone(100 μmol/L),and nuclear factor kappa B(NF-κB) inhibitor primed cells were co-incubated with NTHi for 24 h. Then levels of interleukin-8(IL-8) and tumor necrosis factor-α(TNF-α) in the supernatant was assayed by enzyme-linked immunosorbent assay(ELISA) and the expression of intercellular adhesion molecule-1(ICAM-1) in cells was detected by immunohistochemistry staining. Results A549 cells were transformed and died after co-intubated with NTHi for 24 h. NTHi induced A549 cells to release significantly greater amounts of IL-8, which was inhibited by NF-κB inhibitor pyrrolidine dithiocarbamate(PDTC). Incubating of A549 cells with NTHi significantly induced release of IL-8 and the expression of ICAM-1, which was blocked by erythromycin and dexamethasone and not by gentamicin. TNF-α was not detected in all circumstances. Conclusions NTHi can increase significantly the release and expression of proinflammatory cytokines through NF-κB pathway. Antibacterial drug erythromycin also has anti-inflammatory effect.
Objective To investigate whether long term bronchodilator tiotropium has an acute bronchodilator effect on COPD patients. Methods 46 patients with stable COPD were enrolled in the study.Lung function test was performed before and at 10 min, 20 min, 1 h after inhaling tiotropium. FEV1 , FVC,FEV1/FVC, PEF25% -75% were measured by ambulatory spirometer. The patients were followed up after 1 month.Results The mean FEV1 was ( 1. 110 ±0. 34) L before inhaling tiotropiumand ( 1. 172 ±0. 359) L, ( 1. 221 ±0. 391) L, ( 1. 225 ±0. 392) L at 10 min,20 min, 1 h after inhaling tiotropium, respectively. FEV1 at 1 h after inhaling tiotropiumsignificantly increased compared with that before inhaling tiotropium. FVC also increased and reached highest at 1 h after inhaling tiotropium. PEF25%-75% at 1 h after inhaling tiotropium increased, but there was no significance difference compared with that before inhaling tiotropium. Mean FEV1 was 1. 287 Lafter 1 month, with significant difference compared with baseline. Conclusion Tiotropium can release the symptoms and improve compliance of COPD patients for its acute bronchodilator effect on COPD patients.