OBJECTIVE: To compare the serum glycerophospholipid levels in the inflammatory subtypes of asthma by using targeted metabolomic analysis. METHODS: Demographic and clinical data were collected from 51 patients with asthma between January 2015 and December 2015. Routine blood and sputum induction tests were performed. Eosinophilic asthma was defined as induced sputum containing ⪖ 3% eosinophils, and neutrophilic asthma, as induced sputum containing ⪖ 71% neutrophils. Serum metabolic glycerophospholipid profile was determined by liquid chromatography-mass spectrometry. Differences in glycerophospholipid levels between eosinophilic and non-eosinophilic asthma and between neutrophilic and non-neutrophilic asthma were analyzed using partial least squares discriminant analysis. RESULTS: The serum lysophosphatidylglycerol level was significantly higher in the group with ⪖ 3% eosinophils in sputum than in the group with < 3% eosinophils in sputum. The area under the receiver-operating characteristic curve was ⪖ 70%. There was no significant difference in the serum metabolic glycerophospholipid profile between the group with sputum neutrophils ⪖ 71% and the group with sputum neutrophils < 71%. CONCLUSION Serum lysophosphatidylglycerol is produced abundantly in eosinophilic asthma and may be a biomarker of eosinophilic asthma. This information is helpful for identifying and tailoring treatment for the common asthma subtypes.
Adult ; Asthma ; blood ; immunology ; Eosinophils ; immunology ; Female ; Glycerophospholipids ; blood ; Humans ; Male ; Metabolomics ; Middle Aged ; Neutrophils ; immunology ; Sputum ; cytology ; immunology

Induced sputum and sputum cell count analysis is a test for the diagnosis of various respiratory diseases. In particular, it has long been used as an important biomarker in the diagnosis or characterization of asthma or eosinophilic bronchitis. Despite a relatively long history of this test, there has been no consensus report for conducting and interpreting the analyses in Korea. Based on this awareness and necessity, the Korean Academy of Asthma, Allergy and Clinical Immunology launched the Standardization Committee to review the international guidelines and the literature and to develop a consensus report on the diagnostic procedure and interpretation of the sputum induction test.
Allergy and Immunology ; Asthma ; Bronchitis ; Cell Count ; Consensus ; Diagnosis ; Eosinophils ; Hypersensitivity ; Korea ; Methods ; Sputum*

Tuberculosis has traditionally been confirmed by AFB staining or culturing Mycobacterium tuberculosis from clinical specimens. However, because of the low sensitivity of the sputum smear examination and of the delayed report in culturing, the conventional methods for detection of M. tuberculosis have not been always satisfactory. In an attempt to develop an alternate tool, this study was initiated to produce monoclonal antibodies (MAb) to lipoarabinomannan B (LAM-B) antigen and to use the antibodies in detecting mycobacteria. In this study, five monoclonal antibodies specific to LAM-B were produced; LAM701 (IgG3), LAM138 (IgM), LAM204 (IgM), LAM302 (IgM), and LAM604 (IgM). A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for detecting LAM-B and other mycobacterial antigens using the monoclonal antibodies. With the MAb LAM701, the minimal detectable concentration was 1.0 ng/ml for LAM-B, and 1.0 microgram/ml for M. tuberculosis whole cells, respectively. When 14 clinical specimens proven to contain AFB by smear staining or culture were examined, ten (71.4%) were positive by the sandwich ELISA; in contrast, sputum smear examination gave positive results in only six (42.9%) specimens. Meanwhile, none of 25 specimens with no evidence of AFB were positive by the sandwich ELISA using the MAb LAM701. Although further evaluations are required, this study suggests that the monoclonal antibodies to LAM-B may be useful in detecting mycobacteria from clinical specimens.
Antibodies, Monoclonal/*biosynthesis ; Antigens, Bacterial/*analysis ; Enzyme-Linked Immunosorbent Assay ; Human ; Lipopolysaccharides/chemistry/*immunology ; Mycobacterium tuberculosis/*immunology ; Sputum/*microbiology ; Support, Non-U.S. Gov't ; Tuberculosis/*diagnosis/microbiology

Aged ; Bacteria ; isolation & purification ; Bronchodilator Agents ; therapeutic use ; Cytokines ; analysis ; Forced Expiratory Volume ; Humans ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; immunology ; microbiology ; Sputum ; immunology

It has been suggested that dendritic cells (DCs) are critical antigen presenting cells for eosinophilic airway inflammation in a mouse model of asthma, and cysteinyl leukotrienes may play a role in DC trafficking in asthmatics. We investigated whether the number of DCs is increased in the induced sputum of both atopic and nonatopic asthmatics and is related to activated eosinophil count in the sputum. Sputum was induced by inhalation of hypertonic saline in 9 atopic and 12 nonatopic asthmatics and 10 nonatopic normal controls, and differential cell counts were performed. DCs and activated eosinophils were identified by immunocytochemistry with monoclonal antibodies (anti-CD1a and EG2, respectively). There were significantly higher percentages of eosinophils, EG2+ cells, and CD1a+ DC in the sputum of atopic and nonatopic asthmatics compared with normal controls, respectively. In asthmatics, the percentage of CD1a+ DC was significantly correlated with that of EG2+ cells (Rs=0.62, p=0.004). We demonstrated that the increased number of DCs was evident in the induced sputum of both atopic and nonatopic asthmatics, and the DC number was related to the activated eosinophil count, which suggests that DCs may contribute to the ongoing eosinophilic inflammation in asthmatic airways, and vice versa.
Adult ; Aged ; Antigens, CD1/analysis ; Asthma/*immunology/pathology ; Comparative Study ; Dendritic Cells/*immunology ; Eosinophil Granule Proteins/analysis ; Eosinophils/cytology/*immunology ; Female ; Humans ; Immunohistochemistry ; Leukocyte Count ; Male ; Middle Aged ; Research Support, Non-U.S. Gov't ; Sputum/cytology/*immunology

Corticosteroids are considered to be one of the most effective medicine for asthma by suppressing airway inflammation. This study was carried out to investigate the effects of prednisolone in the sputum of exacerbated asthmatics. Clinical severity, cell differentials, levels of interleukin (IL)-5, eosinophil cationic protein (ECP), EG2+ eosinophils, and nitric oxide (NO) metabolites were measured. Sputum was examined 2 weeks apart in 13 exacerbated asthmatics before and after prednisolone treatment, and once in 12 stable asthmatics. We used a sandwich ELISA for IL-5, fluoroimmunoassay for ECP, immunohistochemical staining for EG2+ eosinophils, a NO metabolites assay using modified Griess reaction. Exacerbated asthmatics, in comparison with stable asthmatics, had significantly higher proportion of eosinophils, higher level of ECP, higher percentage of EG2+ eosinophils, and NO metabolites. Exacerbated asthmatics after treatment with prednisolone had reduced the proportions of eosinophils, reduced level of IL-5, ECP and percentage of EG2+ eosinophils. FEV1 was correlated with the proportion of eosinophils, ECP, and IL-5 respectively. These findings suggest that prednisolone is considered to be effective medicine for asthma by suppressing eosinophil activation through IL-5.
Administration, Oral ; Adolescence ; Adrenal Cortex/metabolism ; Adult ; Aged ; Anti-Inflammatory Agents, Steroidal/administration & dosage* ; Asthma/metabolism ; Asthma/immunology* ; Asthma/drug therapy* ; Biological Markers ; Blood Proteins/metabolism* ; Eosinophils/metabolism ; Eosinophils/immunology ; Eosinophils/drug effects* ; Female ; Human ; Interleukin-5/metabolism* ; Leukocyte Count ; Male ; Middle Age ; Nitric Oxide/metabolism ; Prednisolone/administration & dosage* ; Sputum/immunology ; Sputum/cytology

BACKGROUNDThe airway inflammation could be assessed by some noninvasive approaches. To investigate the value of eosinophil counts in induced sputum and fractional concentration of exhaled nitric oxide (FENO) for the regimen adjustment in patients with asthma, the correlation was analyzed between the two parameters and lung function parameter (forced expiratory volume in one second (FEV(1))).

METHODSSixty-five outpatients with mild to moderate non-exacerbation asthma from Beijing Chao-Yang Hospital were enrolled as treatment group. Combined medications of inhaled corticosteroids plus long-acting beta-2 agonist were administered for one year. Lung function parameters, eosinophil counts in induced sputum, concentration of exhaled nitric oxide and the Asthma Control Test scores were recorded, at regular intervals in the follow-up period. Twenty-one healthy volunteers were enrolled as control group and underwent examination of eosinophil counts in induced sputum, lung function and concentration of exhaled nitric oxide.

RESULTSSixty-three subjects from treatment group completed follow-up period for one year or longer. Mean FEV(1) value of the 63 subjects was (2.75 ± 0.54) L at baseline, (2.97 ± 0.56) L and (3.07 ± 0.52) L at month 3 and month 6, respectively, and maintained as (3.14 ± 0.51) L in the following six months. Mean FENO decreased from (61 ± 25) parts per billion (ppb) at baseline to (32 ± 19) ppb at month 3 (P < 0.05), and continued to decrease to (22 ± 12) ppb at month 6, the difference being significant when compared to both baseline and control group ((13 ± 8) ppb). Mean eosinophil counts decreased to (0.032 ± 0.011) × 10(6)/ml at month 3, which was significantly different from baseline ((0.093 ± 0.023)×10(6)/ml) and the control group ((0.005 ± 0.003)×10(6)/ml (both P < 0.05). The eosinophil counts in induced sputum correlated positively with concentration of FENO in the first six months (all P < 0.05). The concentration of FENO had a significant negative correlation with FEV(1) value (all P < 0.05) in any time point in the follow-up period. The Asthma Control Test scores were 18 ± 5, 19 ± 7, 23 ± 2, 24 ± 1 and 24 ± 1 at months 1, 3, 6, 9 and 12, respectively, which were significantly different from the score at baseline (14 ± 3) (P < 0.05). The most rapid clinical effect was observed at the second month after treatment.

CONCLUSIONEosinophil counts in induced sputum and FENO are sensitive parameters to detect airway inflammation and may be useful in evaluating the efficacy of treatment and adjusting medication regimens.

Adult ; Asthma ; immunology ; physiopathology ; Breath Tests ; Eosinophils ; physiology ; Female ; Humans ; Leukocyte Count ; Lung ; physiopathology ; Male ; Middle Aged ; Nitric Oxide ; analysis ; Sputum ; cytology

BACKGROUNDChronic obstructive pulmonary disease (COPD) is a progressive disease associated with a cellular inflammatory response mostly concerned with cigarette smoking. Chemokine receptors CCR1/5 play an important role in the inflammatory cells recruitment in the lung of COPD patients. The aim of this study was to determine the impact of cigarette smoking on the expression of CCR1/5 on inflammatory cells in induced sputum, and the relationship between the receptors expression and COPD severity.

METHODSDifferential cells in induced sputum were counted and the optical densities of CCR1 and CCR5 on inflammatory cells in induced sputum from COPD patients (n = 29), healthy smokers (n = 11), and nonsmokers (n = 6) were measured using immunocytochemistry. Concentrations of CCL3, the ligand of CCR1/5, in supernatant of induced sputum were detected by enzyme-linked immunosorbent assay.

RESULTSThe expressions of CCR1 and CCR5 on inflammatory cells in healthy smokers were significantly higher than those in nonsmokers, and the expression of CCR1 in patients with COPD was significantly increased when compared with nonsmokers but not healthy smokers. The expressions of CCR1 and CCR5 on inflammatory cells in severe and very severe COPD patients were higher compared with mild and moderate COPD patients. CCL3 level was positively correlated with the total cell counts in induced sputum and smoking history, and negatively correlated with percentage of predicted FEV(1).

CONCLUSIONSCigarette smoking could increase the expression of CCR1 on the inflammatory cells. Both CCR1 and CCR5 expressions on the inflammatory cells in induced sputum could be associated with COPD severity.

Adult ; Aged ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; immunology ; metabolism ; Receptors, CCR1 ; metabolism ; Receptors, CCR5 ; metabolism ; Smoking ; adverse effects ; Sputum ; cytology ; immunology

OBJECTIVETo evaluate the effect of Yufeining, a traditional Chinese medicine, on induced sputum interleukin-8 (IL-8) in patients with chronic obstructive pulmonary disease (COPD) at the stable phase.

METHODSThirty-six patients with COPD were divided into trial group (18 cases) and control group (18 cases) randomly. The trial group was treated with Yufeining pills taken orally for half a year; the control group was not given any medicine. Routine lung function was recorded before and after treatment. Total cell count (TCC), differential cell counts (DCCs) and IL-8 in induced sputum were determined at the baseline and 6 months later.

RESULTSThe indices of lung function improved significantly after 6 months' treatment in trial group (P < 0.05); TCC and absolute neutrophil count decreased significantly compared with baseline in the trial group (P < 0.05); Sputum IL-8 concentration dropped significantly after 6 months' treatment, from a mean of 5.216 +/- 2.914 microg/L to 4.222 +/- 2.140 microg/L (P < 0.05). There were insignificant changes in the parameters in the control group between baseline and 6 months later.

CONCLUSIONYufeining could improve lung function, decrease sputum TCC, absolute neutrophil count and IL-8 concentration, and relieve airway inflammation in patients with COPD in the stable phase.

Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-8 ; analysis ; immunology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; immunology ; Respiratory Function Tests ; Sputum ; chemistry

Adult ; Asthma ; drug therapy ; immunology ; Budesonide ; therapeutic use ; Cell Differentiation ; Eosinophil Cationic Protein ; analysis ; Female ; Humans ; Interleukin-17 ; analysis ; Interleukin-8 ; analysis ; Male ; Middle Aged ; Neutrophils ; physiology ; Peroxidase ; analysis ; Prospective Studies ; Sputum ; cytology ; immunology