Biomarkers ; analysis ; Bronchoalveolar Lavage Fluid ; immunology ; Eosinophils ; immunology ; Humans ; Hypersensitivity ; immunology

OBJECTIVETo study the status of Th1/Th2 immune response and the value of the detection of cytokines in bronchoalveolar lavage fluids (BALF) by examining the levels of IFN-γ and IL-4 in BALF and serum in children with severe Mycoplasma pneumonia (MPP).

METHODSThe levels of IFN-γ and IL-4 in BALF and serum were measured using ELISA in 25 children with severe MPP, 25 children with mild MPP and 25 children with foreign body in bronchus.

RESULTSThe levels of IL-4 and IFN-γ and the IL-4/IFN-γ ratio in BALF in children with severe MPP were significantly higher than those in children with foreign body in bronchus (P<0.01 or P<0.05). The serum levels of IL-4 and the IL-4/IFN-γ ratio in children with severe MPP were significantly higher than those in children with foreign body in bronchus (P<0.01) or with mild MPP (P<0.05). The levels of IL-4 and the IL-4/IFN-γ ratio in BALF were significantly higher than in serum (P<0.05).

CONCLUSIONSThe data suggest that the imbalance of Th1/Th2 exists in children with severe MPP and it seems to represent a predominant Th2-like cytokine response. The detection of cytokines in BALF appears to be more sensitive than in serum and may be of value in the diagnosis and therapy of MPP.

Adolescent ; Bronchoalveolar Lavage Fluid ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Pneumonia, Mycoplasma ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo investigate the effect of Mycoplasma pneumoniae (MP) infection on the function of T lymphocytes in the bronchoalveolar lavage fluid (BALF) of asthmatic children in acute and stable periods and the relationship between MP infection and asthma.

METHODSSeventy-one hospitalized children (with bronchitis, pneumonia, and asthma) were divided into non-MP infection control group (group A, pneumonia and bronchitis without MP infection), non-MP infection asthma group (group B), and MP infection asthma group (group C). Flow cytometry was used to determine CD3(+), CD4(+), and CD8(+) T cell counts and CD4(+)/CD8(+) ratio in BALF among all children in acute and stable periods.

RESULTSCompared with group A, groups B and C showed significant differences in CD3(+), CD4(+), and CD8(+) T cell counts and CD4(+)/CD8(+) ratio (P<0.05) in acute and stable periods, had decreased CD3(+) and CD4(+) T cell counts, an increased CD8(+) T cell count, and a significantly decreased CD4(+)/CD8(+) ratio (P<0.05) in the acute period, and had decreased CD3(+) and CD4(+) T cell counts and CD4(+)/CD8(+) ratio and an increased CD8(+) T cell count (P<0.05) in the stable period. Compared with group B, group C had significantly decreased CD3(+) and CD4(+) T cell counts and CD4(+)/CD8(+) ratio (P<0.05) and a significantly increased CD8(+) T cell count (P<0.05) in the acute period and showed no significant differences in CD3(+), CD4(+), and CD8(+) T cell counts (P>0.05) and a significant decrease in CD4(+)/CD8(+) ratio (P<0.05) in the stable period.

CONCLUSIONSThe immunological function of T lymphocytes in the airway declines significantly among asthmatic children with MP infection in acute and stable periods, leading to immue system disorder. MP may be associated with the pathogenesis of asthma.

Asthma ; etiology ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; CD4-CD8 Ratio ; Child ; Child, Preschool ; Female ; Humans ; Male ; Pneumonia, Mycoplasma ; complications ; immunology ; T-Lymphocytes ; immunology

BACKGROUNDCigarette smoke induces an acute but persisting inflammation in peripheral blood and airway in chronic obstructive pulmonary disease (COPD), and CD8(+) Tc-lymphocytes are considered as a key role in this process. We aimed to investigate the Tc-lymphocytes immunodeviation in system and local airway of COPD patients and changes of the immunodeviation after short-term smoking cessation.

METHODSPeripheral blood (PB) and bronchoalveolar lavage fluid (BALF) were collected from 42 patients (14 COPD patients, 16 smokers with normal lung function and 12 nonsmokers), while PB and induced sputum (IS) were obtained from other 19 patients (10 quitting smokers and 9 continuing smokers) at baseline and follow-up respectively of 4-week smoking cessation. Percentages of CD8(+) Tc-lymphocytes (%CD3(+)) and Tc1/Tc2 ratios were measured by flow cytometry.

RESULTSPercentages of CD8(+) Tc-lymphocytes were higher in COPD patients than those in smokers and nonsmokers in both PB and BALF. Tc1/Tc2 ratio in PB and in BALF from COPD patients was greater than that from smokers and nonsmokers and negatively correlated with FEV1 %pre. When comparing the ratios between PB and BALF, significantly positive correlation was found in COPD patients. Furthermore, after 4-week smoking cessation, percentages of CD8(+) Tc-lymphocytes in PB and IS in quitting smokers were decreased compared to that in baseline and continuing smokers, whereas Tc1/Tc2 ratios were not influenced.

CONCLUSIONSCD8(+) Tc1-trend immunodeviation profiles occurred in both system and local airway of COPD patients. This exceptional immunodeviation could not be relieved by short-term smoking cessation.

Aged ; Bronchoalveolar Lavage Fluid ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Lung ; physiopathology ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; immunology ; physiopathology ; Smoking Cessation ; Time Factors

OBJECTIVETo observe the changes in Th17 cell levels in bronchoalveolar lavage fluid in children of sepsis with acute lung injury and the relationship between the Th17 cell levels and prognosis.

METHODSFifty children of sepsis with acute lung injury were enrolled in the study. The percentages of Th17 cells in bronchoalveolar lavage fluid were measured by flow cytometry. The patients of sepsis with acute lung injury were classified into three groups based on the Pediatric Critical Illness Score (PCIS): extremely critical, critical and non-critical. According to the clinical prognosis, the patients were classified into survical and death groups. Th17 cell levels were compared between the two groups. The relationship between Th17 cell levels and the PCIS scores was analyzed.

RESULTSWith the increase in the severity of sepsis, Th17 cell levels in bronchoalveolar lavage fluid were gradually increased (P<0.05). The Th17 cell levels were negatively correlated to the PCIS scores (r=-0.853; P<0.01). The Th17 cell levels were significantly higher in the death group than in the survival group. Moreover, compared with the survival group, the PCIS scores were lower in the death group (P<0.05).

CONCLUSIONSThe increased Th17 cell levels in children of sepsis with acute lung injury are closely related to the severity and prognosis of patients, suggesting that Th17 cell levels can be used as a predictor of the severity and prognosis.

Acute Lung Injury ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Prognosis ; Sepsis ; immunology ; Severity of Illness Index ; Th17 Cells ; physiology

OBJECTIVETo study the concentrations of IL-4 and IL-13 in bronchoalveolar lavage fluid (BALF) in neonates with respiratory distress syndrome (RDS) and concurrent ventilator-associated pneumonia (VAP).

METHODSSixty-eight neonates with RDS undergoing mechanical ventilation for over 48 hrs were enrolled. IL-4 and IL-13 levels in BALF were measured using ELISA 1, 72 and 96 hrs after mechanical ventilation. The results were compared between the neonates with concurrent VAP (n=37) and without (n=31).

RESULTSThe levels of BALF IL-4 96 hrs after ventilation in the VAP group (35.34+/-1.78 ng/mL) were significantly higher than those in the non-VAP group (13.69+/-2.47 ng/mL, P<0.05). The levels of BALF IL-13 96 hrs after ventilation in the VAP group (33.74+/-2.74 ng/mL) also increased significantly compared with those in the non-VAP group (13.50+/-3.81 ng/mL) (P<0.05). There were significant differences in BALF IL-4 and IL-13 levels between 1 hr and 96 hrs in the VAP group (P<0.05).

CONCLUSIONSBALF IL-4 and IL-13 levels increase in neonates with RDS and concurrent VAP. IL-4 and IL-13 may involve in the regulation of the inflammatory immune response.

Bronchoalveolar Lavage Fluid ; immunology ; Female ; Humans ; Infant, Newborn ; Interleukin-13 ; analysis ; Interleukin-4 ; analysis ; Male ; Microbial Sensitivity Tests ; Pneumonia, Ventilator-Associated ; immunology ; microbiology ; Respiratory Distress Syndrome, Adult ; immunology

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 ; Asthma/*immunology ; Bronchoalveolar Lavage Fluid/chemistry/cytology/immunology ; Cytokines/biosynthesis/immunology ; Disease Models, Animal ; Female ; Immunization ; Immunomodulation/*immunology ; Inflammation/*immunology ; Leukocytes/immunology ; Macrophages, Alveolar/*immunology ; Mice ; Mice, Inbred C57BL ; Ovalbumin/immunology

OBJECTIVEPrevious studies have shown that bacillus calmette-guerin (BCG) can deviate TH2 response toward TH1 response, resulting in a suppressive effect on the development of asthma/atopy. This study examined the effect of BCG treatment on regulatory T cells in asthmatic mice to investigate the possible mechanism.

METHODSKunming mice were sensitized and challenged with ovalbumin (OVA) to establish asthmatic models. Asthmatic mice were injected intradermally with BCG five days before and after sensitization. After 24 hrs of last challenge, bronchoaveolar lavage fluid (BALF) and peripheral blood were collected . The total cells and eosinophils were counted in the BALF. The percentage of CD4(+) CD25(+) in peripheral blood was detected with flow cytometry. Single spleen cell suspension was prepared and cultured in 1640 medium for 48 hrs and then the cytokine IL-10 level in the supernatant was determined using ELISA. The mice which were challenged with normal saline were used as the Normal control group.

RESULTSThe number of total cells and eosinophils in BALF in asthmatic mice [(27.27 +/- 5.36) x 10(7)/L and (6.59 +/- 1.32) x 10(7)/L respectively] were more than in the Normal control group [(1.52 +/- 0.36) x 10(7)/L and zero respectively] (P < 0.01). The number of total cells and eosinophils in BALF in asthmatic mice were reduced after BCG treatment [(13.71 +/- 3.17) x 10(7)/L and (1.43 +/- 0.37) x 10(7)/L respectively] (P < 0.01). The percentage of CD4(+) CD25(+) in peripheral blood of asthmatic mice [(11.59 +/- 1.33)%] was noticeably lower than that of the Control group [(13.66 +/- 1.68)%] (P < 0.01), but increased significantly in asthmatic mice after BCG treatment [(14.40 +/- 2.70)%] (P < 0.05). The IL-10 level in spleen cell supernatant in the BCG-treated group (7.79 +/- 1.34 pg/mL) also increased compared with that in the untreated asthmatic mice (5.54 +/- 0.66 pg/mL) (P < 0.01).

CONCLUSIONSBCG can markedly inhibit the airway inflammation in asthmatic mice possibly by promoting the production of regulatory T cells.

Animals ; Asthma ; immunology ; therapy ; BCG Vaccine ; therapeutic use ; Bronchoalveolar Lavage Fluid ; cytology ; Interleukin-10 ; analysis ; physiology ; Male ; Mice ; T-Lymphocytes, Regulatory ; immunology ; Toll-Like Receptor 2 ; physiology

OBJECTIVETo measure levels of interleukin-4 (IL-4) and interferon-gamma (INF-γ) in the bronchoalveolar lavage fluid (BALF) of children with refractory Mycoplasma pneumoniae pneumonia (RMPP), and to investigate changes in local Th1-Th2-type cytokine levels in children with RMPP and their significance.

METHODSA total of 42 children with RMPP were divided into atopic (n=11) and non-atopic groups (n=31) according to whether they had eczema, allergic rhinitis, urticaria, and family history of allergic disease. The study also included a control group of 12 children with bronchial foreign bodies who underwent foreign body removal and were re-examined by fiberoptic bronchoscopy four weeks later. The different cells in BALF from all children were analyzed, and the levels of IL-4 and INF-γ in BALF were measured using enzyme-linked immunosorbent assay.

RESULTSCompared with the control group, the total number of cells in BALF from children with RMPP increased significantly (P<0.05), the increase mainly accounted for by neutrophils (P<0.01), and levels of IL-4 and INF-γ in BALF from children with RMPP increased significantly (P<0.05). Compared with the control group, levels of IL-4 and INF-γ in BALF in the atopic group increased significantly (P<0.05). The level of INF-γ in BALF in the non-atopic group also increased significantly (P<0.05). There were no significant differences in INF-γ/IL-4 ratio among all groups (P>0.05).

CONCLUSIONSSignificant increase in cell numbers, especially neutrophils, as well as IL-4 and INF-γ levels, can be seen in BALF from children with RMPP, but there is no change to the INF-γ/IL-4 ratio. This indicates a significant local inflammatory response in children with RMPP, but there is no evidence of Th2-dominated inflammatory response.

Adolescent ; Bronchoalveolar Lavage Fluid ; cytology ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Interferon-gamma ; analysis ; Interleukin-4 ; analysis ; Male ; Pneumonia, Mycoplasma ; immunology

BACKGROUNDAsthma is a chronic airway disease with inflammation characterized by physiological changes (airway hyper-responsiveness, AHR) and pathological changes (inflammatory cells infiltration and mucus production). Eosinophils play a key role in the allergic inflammation. But the causative relationship between eosinophils and airway inflammation is hard to prove. One of the reasons is lack of activation marker of murine eosinophils. We investigated the expression of CD69 on murine eosinophils in vitro, the relationship between the expression of CD69 on eosinophils from peripheral blood and bronchoalveolar lavage fluid and on airway inflammation in asthmatic mice.

METHODSEosinophils from peripheral blood of IL-5 transgenic mice (NJ.1638) were purified. Mice were divided into five groups: wild type mice sensitized and challenged with saline (WS group), wild type mice sensitized and challenged with ovalbumin (WO group), IL-5(-/-) mice sensitized and challenged with saline and transferred with purified eosinophils (ISE group), IL-5(-/-) mice sensitized and challenged with OVA and transferred with purified eosinophils (IOE group), IL-5(-/-) mice sensitized and challenged with OVA and transferred with purified eosinophils, pretreated with anti CD4 monoclonal antibody (IOE+antiCD4mAb group). IL-5(-/-) mice were sensitized with OVA at day 0 and day 14, then challenged with OVA aerosol. On days 24, 25, 26 and 27 purified eosinophils were transferred intratracheally to IL-5(-/-) mice. On day 28, blood and BALF were collected and CD69 expression on eosinophils measured by flowcytometry.

RESULTSPurified eosinophils did not express CD69. But eosinophils cultured with PMA + MA, IFN-gamma, IL-5 or GM-CSF expressed CD69 strongly. Eosinophils from blood of WO, WS group did not express CD69 at all. The numbers of eosinophils in BALF of WO group, IOE group, ISE group and IOE + antiCD4mAb group were significantly higher than in mice of WS group which did not have eosinophils at all. CD69 expression on eosinophils in BALF of IOE and WO groups was strong. Eosinophils in BALF of ISE and IOE + antiCDmAb groups did not express CD69. The mucus production result was similar to CD69 expression. There were eosinophils infiltration in lung slides of all groups except WS group.

CONCLUSIONActivation in airway of eosinophils could directly lead to airway inflammation.

Animals ; Antigens, CD ; analysis ; Antigens, Differentiation, T-Lymphocyte ; analysis ; Asthma ; immunology ; physiopathology ; Bronchoalveolar Lavage Fluid ; cytology ; Eosinophils ; immunology ; Inflammation ; physiopathology ; Lectins, C-Type ; Lung ; physiopathology ; Mice ; Mice, Transgenic