Relationship between the perception of dyspnoea and airway inflammatory markers

Poor dyspnoea perception in asthmatic patients seems to be associated with increased risk of asthma exacerbation. We have studied the relationship between basel ne dyspnoea perception and inflammatory markers in sputum in eight patients with mild asthma and in 13 patients with moderate to severe asthma.The perception of dyspnoea was scored on the Borg scale. Eosinophilic cationic protein (ECP) was measured by fluoroimmunoassay and by an interleukin (IL)-5 sandwich ELISA. The baseline Borg score was significantly higher in patients with severe asthma than in patients with mild to moderate asthma (4.1 +/- 0.29 vs. 2.28 +/- 0.28, P<0.05).The proportion of eosinophil and ECP levels in the sputum were significantly higher in patients with moderate to severe asthma. IL-5 in sputum was significantly increased in moderate to severe asthmatic patients compared to mild asthmatic patients. A significant relationship was found between the baseline perception score and FEV1/FVC (r = -0.53, P<0.01), sputum eosinophils (r = 0.70, P<0.01) and sputum ECP (r = 0.62, P<0.01).These findings suggest that the baseline perception score is related to inflammatory markers in sputum, and that the perception of dyspnoea as well as airway inflammatory markers may be considered to evaluate asthma severity.     

NO in exhaled air is correlated with markers of eosinophilic airway inflammation in corticosteroid-dependent childhood asthma.

The relationship between nitric oxide in exhaled air, levels of sputum eosinophils, sputum eosinophil cationic protein (ECP) and urinary eosinophil protein X (EPX) excretion has not yet been investigated in corticosteroid-dependent childhood asthma. Therefore, taking 25 children with stable asthma (mean age 11.2 yrs) treated with inhaled corticosteroids and nine nonatopic healthy control children (mean age 12.8 yrs) the level of exhaled NO was measured by means of a chemiluminescence analyser before and after sputum induction. This was conducted as a slow vital capacity manoeuvre under standardized conditions with a target flow of 70 mL x s(-1) against a resistance of 100 cm H2O x L(-1) x s. Sputum induction was performed by inhalation of hypertonic saline (3, 4, and 5%) in a standardized manner and a single sample of urine was collected. Exhaled NO (p = 0.01), absolute eosinophil cell counts in sputum (p = 0.02), sputum ECP (p = 0.09) and urinary EPX excretion (p = 0.02) were higher in asthmatics compared to control children. Exhaled NO was positively correlated with sputum ECP (r(s) = 0.59, p = 0.002), urinary EPX (r(s) = 0.42, p = 0.03), and sputum eosinophils (r(s) = 0.30, p = 0.15) in the asthmatic children. These correlations appeared to be pronounced after sputum induction, where NO values had decreased (p = 0.01). None of the correlations were observed in the group of nonatopic control subjects. Additionally there were significant correlations between sputum ECP and sputum eosinophils (r(s) = 0.69, p<0.001) as well as between sputum ECP and urinary EPX excretion (r(s) = 0.58, p = 0.003) in the asthmatics. Exhaled NO provides information about the degree of eosinophilic airway inflammation and thus appears to be a useful and easy-to-perform inflammatory marker in corticosteroid-dependent asthma.     

Eotaxin level in induced sputum is increased in patients with bronchial asthma and in smokers

BACKGROUND: Airway eosinophilia is one of the hallmarks of asthma. Eotaxin may play an important role in eosinophil recruitment. OBJECTIVES: To examine the relationship between eotaxin levels in the sputum and eosinophilic inflammation. METHODS: The sputum was obtained from 11 non-smokers, 14 smokers and 13 asthmatic patients using a sputum induction method. Eotaxin and interleukin (IL)-5 levels in the sputum were determined by ELISA and immunocytochemical analysis. RESULTS: Asthmatic patients had eosinophilia and smokers showed neutrophilia in their sputum. The eotaxin level in the sputum was significantly higher in smokers (median 412.5, range 91.1-872.2 pg/ml) and asthmatic patients (351.0, 185.0-928.0 pg/ml) compared with non-smokers (123.2, 0-369.0 pg/ml; both p < 0.05). IL-5 was detected in the sputum of 1 non-smoker, none of the smokers and 4 asthmatic patients. The percentage of eotaxin-positive cells was higher in smokers and asthmatic patients than in non-smokers, but the percentage of IL-5-positive cells was significantly higher only in asthmatic patients (p < 0.05). CONCLUSIONS: These findings suggest that the elevated eotaxin level in the sputum does not always accompany the increase in eosinophils, and cooperation with another cytokine such as IL-5 may be required for the recruitment of eosinophils.     

Exhaled nitric oxide and sputum eosinophil markers of inflammation in asthmatic children.

Exhaled nitric oxide and eosinophil sputum markers are considered noninvasive ways in which to evaluate airway inflammation in asthma. The aim of this study was to evaluate the relationships between these methods of evaluation in asthmatic children. In a cross-sectional study of 25 mild-moderate asthmatic children (aged 6-13 yrs, 10 patients on inhaled steroids) exhaled NO was measured along with induced sputum by inhalation of hypertonic saline solution. The sputum was processed for eosinophil count and eosinophil cationic protein (ECP) determination. Serum ECP and lung function (forced expiratory volume in one second (FEV1)) were also measured. A significant correlation was observed between exhaled NO and sputum eosinophils (r = 0.438, p = 0.032) as well as between sputum eosinophils and sputum ECP (r = 0.532, p<0.01). No correlation was observed among exhaled NO and serum ECP, sputum ECP, FEV1, respectively. Furthermore no correlation was observed between sputum eosinophil (%) and serum ECP and between sputum eosinophils and FEV1. There was no correlation among the investigated parameters in children treated with inhaled steroids. In conclusion, exhaled NO and sputum eosinophil counts are concordant in evaluating the degree of airway inflammation in patients with mild-to-moderate asthma. However, the association between these two noninvasive markers becomes less in steroid treated patients.     

Kinetics of allergen-induced airway eosinophilic cytokine production and airway inflammation.

Airway eosinophilia is the hallmark of asthma exacerbation. Coordination of cytokines such as interleukin (IL)-5, eotaxin and regulated on activation, normal T-cell expressed and secreted (RANTES) seem to be necessary for eosinophil extravasation including adhesion, chemotaxis, and activation. The purpose of this study was to characterize both the kinetics of allergen-induced inflammatory cell recruitment to the airways and cytokines selective for eosinophil chemotaxis, activation, or resolution. Eight atopic asthmatic individuals demonstrating a dual response to inhaled allergen completed a diluent-controlled crossover study. The subjects showed significant allergen-induced early and late airway asthmatic responses (p < 0.001), and an increase in the number of sputum eosinophils and metachromatic cells (p < 0.05). The number of eosinophils immunopositive for IL-5, eotaxin, and RANTES increased 7 h after allergen inhalation (p < 0.05), coincident with the peak number of activated eosinophils. Sputum cells immunopositive for IL-10 decreased significantly following allergen challenge (p = 0. 04), and correlated negatively with sputum eosinophils (r = -0.34, p = 0.02). This study shows that allergen-induced increases in sputum eosinophils are associated with the presence of cytokines specific for the activation and chemotaxis of eosinophils, and suggests that cooperation of eosinophilic cytokines may be important for the accumulation and regulation of activated eosinophils at the site of allergic inflammation.     

哮喘患者痰液中炎症介质和细胞因子与气道重塑的关系研究

目的　以支气管粘膜网状基底膜厚度作为气道重塑的指标 ,探讨哮喘患者气道重塑与痰液中细胞因子和炎症介质的关系。方法　对 2 0例哮喘患者和 10名正常对照者经纤维支气管镜行(纤支镜 )支气管粘膜活检 ,测量支气管粘膜网状基底膜厚度 ,用荧光酶联免疫方法测定痰液中的嗜酸细胞阳离子蛋白 (ECP) ,酶联免疫法测定白细胞介素 5 (IL 5 )、肿瘤坏死因子 (TNF α)的水平 ;采用SPSS8 0统计软件作等级相关分析 ,探讨哮喘患者痰液中ECP、IL 5、TNF α水平与支气管粘膜厚度的关系。结果　缓解期哮喘患者支气管粘膜网状基底膜厚度为 (10 1± 2 6 ) μm ,与正常对照组 [(4 4± 1 2 )μm]比较 ,差异有显著性 (P <0 0 0 5 ) ;哮喘组痰液中ECP水平为 (14 4± 80 ) μg/L、IL 5为 (17± 4 ) μg/L、TNF α为 (14 6± 79) μg/L ,与正常对照组 [(81± 4 4 ) μg/L、(14± 4 ) μg/L、(5 3± 36 ) μg/L]比较 ,差异有显著性 (P <0 0 0 5 ) ;两组痰液中ECP、IL 5与支气管粘膜厚度呈明显正相关 (r =0 5 6 9、0 4 6 6 ,P均 <0 0 0 5 ) ;两组痰液中TNF α水平与粘膜厚度无明显相关 (r=0 2 5 4 ,P >0 0 5 )。结论　缓解期哮喘患者支气管粘膜网状基底膜厚度均存在不同程度增厚 ;而痰液ECP和IL 5水平与支气管粘膜厚度呈     

诱导痰炎性标志物检测在判定哮喘严重程度和鉴别诊断中的应用

目的探讨哮喘患者诱导痰中嗜酸性粒细胞(Eos)和嗜酸细胞阳离子蛋白(ECP)与其哮喘严重程度的关系及鉴别诊断价值。方法选择武汉大学人民医院2002-07~2004-06的门诊哮喘患者59例,检测其肺功能并分别采用瑞氏染色及荧光免疫法检测高渗盐水诱导痰中Eos数量和ECP质量浓度。选择同期20例慢性阻塞性肺疾病患者和10名健康人作为对照。结果哮喘患者诱导痰中Eos数量与患者第1秒用力呼气容积/用力肺活量(FEV1%)呈显著负相关(r=-0·65,P<0·01);ECP质量浓度与患者FEV1%呈显著负相关(r=-0·59,P<0·01)。随病情加重哮喘患者诱导痰中Eos数量和ECP质量浓度逐渐升高,且轻度、中度和重度患者之间比较差异有统计学意义(P<0·05)。哮喘患者诱导痰中Eos数量和ECP质量浓度显著高于慢性阻塞性肺疾病组和健康组(P<0·01)。结论诱导痰中Eos和ECP质量浓度可了解哮喘的严重程度,并有助于与慢性阻塞性肺疾病的鉴别。     

One-year evaluation of the preventative effect of hydrofluoroalkane-beclomethasone dipropionate on eosinophilic inflammation of asthmatic peripheral airways

BACKGROUND: In asthmatic patients with eosinophilic inflammation of the peripheral airways, appropriate drug delivery to the affected area is required. OBJECTIVE: It was the aim of this study to assess persistent eosinophilic inflammation of the peripheral airways in asthmatic patients, stabilized by the long-term use of dry powder type inhaled steroids, and to evaluate the clinical efficacy of hydrofluoroalkane-beclomethasone dipropionate (HFA-BDP) over 1 year. METHODS: Seventy-four outpatients with moderate stable asthma were studied for at least 6 months, 37 treated with fluticasone propionate Diskus (FP-DK) and 37 with budesonide Turbuhaler (BUD-TH). The eosinophil count, eosinophil cationic protein (ECP), eotaxin and RANTES levels in 10% hypertonic saline-induced sputum were examined before treatment, as well as 4 weeks, 8 weeks, 6 months and 1 year after switching patients to HFA-BDP. RESULTS: Fifteen patients (40.5%) in the FP-DK group and 12 (32.4%) in the BUD-TH group had eosinophils in induced sputum. The sputum ECP in the eosinophil-positive and the eosinophil-negative groups was 1,510.1 +/- 2,009.3 versus 426.6 +/- 464.1 microg/l (p = 0.037) in the FP-DK group, and 3,850.0 +/- 5,486.2 versus 492.0 +/- 1,150.7 microg/l (p = 0.011) in the BUD-TH group, respectively. Four weeks after the switch to HFA-BDP, the number of eosinophil-positive patients decreased in both groups. Significant reductions in sputum ECP and eotaxin were observed at 8 weeks, and their concentrations continued decreasing for 1 year. CONCLUSION: There is a certain proportion of asthmatic patients for whom long-term treatment with dry powder type steroids may not be suitable; however, their peripheral airway inflammation improved after switching them to HFA-BDP, suggesting its excellent delivery.     

支气管哮喘气道炎症可能机制的探讨

目的 探讨支气管哮喘(简称哮喘)患者气道炎症特征及其可能机制,并进一步观察吸入糖皮质激素治疗对气道炎性细胞分类计数、炎症介质等的影响.方法 分别选择轻度(轻度组)、中度(中度组)和重度(重度组)持续哮喘患者15例、14例和19例,正常对照组15名,分别行哮喘症状控制评分、肺功能测定、诱导痰炎性细胞分类计数、调节激活正常T细胞表达和分泌细胞因子(RANTES)、嗜酸粒细胞阳离子蛋白(ECP)、白介素8(IL-8)及髓过氧化物酶(MPO)浓度检测,然后规范吸人糖皮质激素治疗4周,随访复查上述指标.结果 诱导痰NEU%、IL-8及MPO重度组明显升高,分别为(62.40±22.05)%、594.53±85.11、39.25±10.67与轻度组[(47.23±15.12)%、183.63±120.98、12.47±4.15]、中度组[(46.13±19.23)%、352.76±71.72、22.93±7.35]、正常对照组[(31.44±13.31)%、103.26±36.33、10.22±4.13]比较差异均有统计学意义(P＜0.01);RANTES、嗜酸粒细胞百分比(EOS%)和ECP浓度在各哮喘组间比较差异无统计学意义(P＞0.05).EOS%与RANTES、ECP水平呈正相关(r=0.557,P＜0.05;r=0.852,P＜0.01);NEU%与IL-8、MPO水平呈正相关(r=0.732,P＜0.05;r=0.806,P＜0.05);经糖皮质激素治疗后,对轻、中、重度哮喘患者合并进行分析表明,治疗后症状评分由(9.8±5.4)分下降至(4.0±3.5)分和肺功能指标第一秒用力呼气容积占预计值百分比由(62.2±23.3)%升高至(75.9±17.5)%显著改善,差异有统计学意义(P＜0.01).在接受糖皮质激素治疗后,RANTES、EOS%和ECP水平均显著降低.另外MPO水平也显著降低(P＜0.01);但治疗后在重度组仍显著高于轻、中度组(P＜0.01).但IL-8、NEU%治疗后无明显降低(P＞0.05),而且治疗后IL-8、NEU%在重度组仍显著高于轻、中度组(P＜0.01).结论 中性粒细胞增多是重度哮喘的气道炎症特征之一,EOS与病情严重程度无关.EOS哮喘的发生可能与RANTES的趋化、EOS的活化、ECP的释放有关,激素可以抑制EOS气道炎症.而中性粒细胞哮喘的发生可能与IL-8的趋化、NEU的活化、MPO的释放有关.     

Clinical Implication of Protein Levels of IL-5 in Induced Sputum in Asthmatic Patients

To determine whether protein levels of interleukin-5 (IL-5) in induced sputum reflect the degree of eosinophilic inflammation, we evaluated the role of IL-5 on clinical characteristics in stable asthmatic patients. IL-5 level, differential eosinophil count, and level of eosinophil cationic protein (ECP) in induced sputum were all significantly higher for asthmatics than for normal controls. Both eosinophil counts and ECP levels in induced sputum were inversely correlated with the degree of airflow limitation (FEV₁/FVC). In addition, patients with measurable IL-5 in sputum had significantly more eosinophils, higher levels of ECP in sputum, and lower FEV₁ (percent predicted) than did patients with levels of IL-5 beneath the limit of detection. However, we found no significant difference in IL-5 levels between atopic and nonatopic asthmatics. IL-5 level in induced sputum is a good indicator of eosinophilic inflammation in atopic and nonatopic asthmatic patients.     

Immunohistochemically stained activated eosinophils in sputum in patients with asthma

BACKGROUND: Eosinophils play an important role in asthmatic airway inflammation. Monoclonal antibody EG2 has been considered to identify activated eosinophils. OBJECTIVE: The present study was aimed to investigate whether immunohistochemically stained EG2+ eosinophils in sputum reflect the severity of asthma. METHODS: Sputum was obtained in 23 asthmatic patients, of whom 13 patients were examined before and after antiasthma treatment including steroid preparations. We used immunohistochemical staining to detect EG2+ (activation marker) eosinophils and fluoroimmunoassay to detect eosinophil cationic protein (ECP). RESULTS: Moderate to severe asthmatics had a significantly higher proportion of eosinophils and EG2+ eosinophils and higher levels of ECP compared to mild asthmatics (40.9 +/- 5.8 vs. 6.4 +/- 1.2%, 35.5 +/- 5.6 vs. 2.7 +/- 1.0%, 1.470.2 +/- 251.5 vs. 210.6 +/- 52.0 microgram/l, respectively; p < 0.01). Significant increases in proportions of eosinophils, EG2+ eosinophils and ECP in the sputum from patients with exacerbated asthma were evident. The proportions of eosinophils, EG2+ eosinophils, and the levels of ECP were reduced following treatment with antiasthmatic drugs. FEV(1) and FEV(1)/FVC were significantly correlated with EG2+ eosinophils. CONCLUSION: These findings demonstrate that EG2+ eosinophils in sputum are closely related to the clinical status in patients with asthma.     

Reduced eosinophil apoptosis in induced sputum correlates with asthma severity.

This study was carried out to investigate the relationship between induced sputum eosinophil apoptosis and clinical severity score, airway obstruction and symptom scores in patients with chronic stable asthma. Altogether, 41 chronic stable asthmatic subjects of varying severity defined by Aas score and 17 control subjects underwent spirometry, symptom questionnaire and successful sputum induction. Sputum was processed and cytospins prepared for light microscopy to determine normal and apoptotic eosinophils. Mild asthmatic subjects had a significantly lower percentage sputum eosinophils and a significantly higher eosinophil apoptotic ratio (AR) than moderate or chronic severe asthmatics. Severe asthmatic subjects had a significantly greater age, duration of asthma and sputum eosinophil count x mL(-1) than mild asthmatic subjects. Asthmatic subjects' symptom scores, severity scores and age inversely correlated with AR and the percentage of sputum eosinophils. Baseline forced expiratory volume in one second inversely correlated with percentage sputum eosinophils and positively correlated with AR. The study demonstrates a relationship between reduced sputum eosinophil apoptosis and increased clinical severity of chronic stable asthma, providing additional evidence that eosinophil apoptosis may be important in the resolution of eosinophilic airway inflammation in asthma.     

Nitric oxide metabolites in patients with asthma: induced sputum versus blood

Nitric oxide (NO) plays an important role in physiological regulation of the airways. The monitoring of airway inflammation has being observed in bronchial asthma directly, by sputum examination, and indirectly, by measurements in peripheral blood. To investigate the diagnostic value of these two methods, we compared NO metabolites in induced sputum and serum obtained in patients with asthma and control subjects. Hypertonic saline induced sputum and serum were obtained in 13 patients with asthma and 10 control subjects. NO metabolite level was assayed by using modified Griess reaction. Eosinophil cationic protein (ECP) was measured by fluoroimmunoassay, and detected interleukin (IL)-5 by a sandwich ELISA. The accuracy of the tests was measured by plotting the data in receiver operating characteristic (ROC) curves and comparing the area under the curve for NO metabolites. Asthmatic patients, compared with control subjects, had significantly higher NO metabolites in induced sputum (1252.5+/-203.3 mol l(-1) vs. 557.2+/-101.5 mol l(-1), P<0.01) but not in serum. IL-5 in induced sputum was detected more frequently in patients with asthma than in control subjects [11/13 (84.6%) vs. 1/10 (10%), P<0.01]. Asthmatic patients, compared with control subjects, had significantly higher ECP concentration in induced sputum (1270.0+/-197.9 g l vs. 154.6+/-47.4 g l(-1), P<0.01). There were significant positive correlations between NO metabolites in induced sputum and eosinophils, ECP in induced sputum (r=0.58 P<0.05; r=0.64, P<0.01) in patients with asthma but not in serum. The area under the ROC curve showed that NO metabolites in induced sputum (0.78) are more accurate marker than NO metabolites in serum (0.53) (P<0.05). These findings suggest that NO metabolites in induced sputum is a more valuable indicator to monitor asthmatic airway inflammation than those in serum.     

哮喘和慢性阻塞性肺疾病患者糖皮质激素治疗前后痰液炎性标志物 …

探讨哮喘和慢性阻塞性疾病患者吸入糖皮质激素治疗后痰液中细菌因子和嗜酸细胞阳离子蛋白浓度及糖皮质激素对其影响。方法采用荧光酶联免疫法检测糖皮质激素治疗前后痰液中白细胞介素（ＩＬ）－５、ＩＬ－８、ＥＣＰ浓度及嗜酸细胞和嗜中性粒细胞计数。     

不同控制水平的支气管哮喘患者气道炎症与外周气道功能状态的研究

目的 了解不同临床控制水平的支气管哮喘(简称哮喘)患者的气道炎症状况及外周气道功能,观察哮喘患者诱导痰中的炎症指标能否反映外周气道功能的改变.方法 收集在北京朝阳医院就诊的哮喘患者66例,分为控制组(21例)、部分控制组(28例)、未控制组(17例)以及健康对照组(20名).所有受试者第1天进行哮喘控制测试(ACT)评分,行脉冲振荡肺功能检测气道阻力及肺功能基础值、诱导痰细胞计数和分类以及嗜酸粒细胞阳离子蛋白(ECP)浓度测定;第2天测定呼出气一氧化氮浓度(FE_(NO)),若所测得的FEV_1≥70%预测值则行乙酰甲胆碱激发试验,当气道阻力升高至基础阻力2倍或乙酰甲胆碱达到最大浓度时终止试验,3 min后行气道阻力及通气功能检测;然后嘱受试者行5次深吸气后再复测气道阻力及通气功能.比较4组受试者诱导痰细胞计数和分类、诱导痰中ECP浓度、FE_(NO)水平与ACT评分间的相关性;观察激发后以及深吸气后外周气道阻力的变化与ACT评分、诱导痰中嗜酸粒细胞(EOS)计数、痰ECP水平及FE_(NO)间的关系.结果 (1)哮喘患者诱导痰EOS计数、ECP浓度以及FE_(NO)随着控制水平的下降逐渐增高,且诱导痰EOS计数、ECP浓度均与ACT评分呈负相关(r值分别为-0.43和-0.56,均P<0.01).(2)在健康对照组,乙酰甲胆碱激发后中心气道阻力(R_(20))、外周气道阻力(R_5-R_(20))增高程度间比较差异无统计学意义(F=3.472,P>0.05),而在控制组及部分控制组激发试验后外周气道的反应强于中心气道(F值分别为18.09和14.14,均P<0.01),但激发后R_5-R_(20)的变化与ACT评分、诱导痰EOS计数、ECP、FE_(NO)水平间无相关性.(3)深吸气后,健康对照组R_5-R_(20)由(0.13±0.14)kPa·L~(-1)·s~(-1)降至(0.08±0.09)kPa·L~(-1)·s~(-1)(t=2.84,P<0.05),而控制组、部分控制组R_5-R_(20)分别由(0.24±0.15)、(0.31±0.18)kPa·L~(-1)·s~(-1)>增至(0.30±0.16)、(0.39±0.17)kPa·L~(-1)·s~(-1)(t值分别为3.90、4.68,均P<0.01),但相关分析显示,深吸气后R_5-R_(20)的变化与ACT评分、诱导痰EOS计数、ECP、FE_(NO)水平无相关性.结论 即使在控制水平的哮喘患者,仍存在气道嗜酸粒细胞炎症,且该炎症状态随着疾病控制水平的降低而逐渐加重;哮喘患者深吸气所致的外周气道舒张作用消失;检测诱导痰中的炎症指标并不能反映外周气道功能的改变.     

Smoking Affects Eotaxin Levels in Asthma Patients

Background. Chronic airway inflammation is most important pathological finding in asthma. Cigarette smoking may modify type of inflammation as well as may influence disease severity and response to the treatment. Objective. Thus the aim of this study was to investigate whether cigarette smoking may have an influence on the levels of eotaxin-1, eotaxin-2, eotaxin-3 and IL-5 in patients with stable mild/moderate asthma. Methods. 45 steroid naive asthmatics (mean age: 55.2 ± 2.2 yrs) and 23 “healthy” smokers and non-smokers control subjects (mean age: 54.4 ± 9.7 yrs) were investigated. Asthmatics were divided into two subgroups according to their smoking histories: asthmatic smokers (n = 19) who currently smoke and have a history of > 10 pack-years and asthmatic never-smokers (n = 26). BAL and induced sputum were performed. Cytospins of induced sputum and BAL were stained with May-Grünwald-Giemsa for differential cell counts. Eotaxin-1, eotaxin-2, eotaxin-3 and IL-5 concentrations in serum, sputum and BAL supernatant was measured using a commercial ELISA kit. Results. In sputum supernatant from asthma smokers was significantly higher concentration of eotaxin-1 than in non-smokers asthmatics (203.4 ± 10.0 vs. 140.2 ± 9.5 respectively, p < 0.05). In non-smokers asthma patients levels of BAL eotaxin-1 strongly related to percent and absolute numbers of BAL eosinophils and neutrophils (Rs = 0.737 and Rs = 0.514 respectively, p < 0.05). The number and percent of sputum neutrophils and eosinophils, obtained from smokers asthmatics, significantly correlated with eotaxin-2 concentration in sputum supernatant (Rs = 0.58 and Rs = 0.75 respectively, p < 0.05). IL-5 levels in the serum and sputum from asthmatic never-smokers were significantly higher than they were from asthmatic smokers and “healthy” smokers. Asthmatic never-smokers showed a significantly higher amount of IL-5 in serum and sputum than the asthmatic smokers showed. Conclusions. This study showed the elevated levels of sputum eotaxin-1 as well as serum, sputum and BAL eotaxin-2 in asthmatic smokers without a significant increase of eosinophils compared to asthmatic never-smokers. The eotaxin concentrations were related not only with number of eosinophils but also with the number of neutrophils in all the studied tissue compartments. The data herein permits a suggestion that smoking may influence change in asthmatic airway inflammation by stimulating the production of eotaxins.     

Eosinophil soluble protein levels, eosinophil peroxidase and eosinophil cationic protein in asthmatic patients.

Eosinophil granular proteins are useful eosinophil activation markers in asthmatic patients. In this study, eosinophil peroxidase (EPO) and eosinophil cationic protein (ECP) levels were assessed in different stages of bronchial asthma in 123 patients suffering from intrinsic (n = 42) and extrinsic (n = 81) asthma, with the aim of evaluating the difference in the protein levels between both types of asthma and their importance as a severity marker of the disease. The geometric mean serum level of EPO was 12.3 +/- 2.17 ng/ml (mean +/- SD) in controls, and 38.6 +/- 3.4 ng/ml in the asthmatic patients. Mean ECP levels were 13.22 +/- 1.11 ng/ml in controls and 30.5 +/- 2.38 ng/ml in patients. Depending on the asthma severity, the EPO levels were 30.4 +/- 4.35, 38.7 +/- 5.29, and 54.46 +/- 9.46 ng/ml in mild, moderate and severe asthmatics, respectively, with the differences being significant between the groups of patients with mild and severe asthma (p < 0.001). ECP levels were 24.23 +/- 3.37 in mild, 31.69 +/- 4.21 in moderate, and 37.61 +/- 4.52 ng/ml in severe asthma. There were significant differences in ECP levels between mild and moderate asthma (p < 0.001) and between mild and severe asthma (p < 0.001). Peripheral eosinophil count was 157 +/- 20 eosinophils/mm3 in controls, 334 +/- 35 eosinophils/mm3 in mild asthmatics, 510 +/- 87 eosinophils/mm3 in moderate asthmatics and 658 +/- 72 eosinophil/mm3 in severe asthmatics, with significant differences between all groups (p < 0.05-p < 0.001). Serum EPO and ECP levels and peripheral eosinophil count were significantly greater in patients with active asthma than in patients with silent asthma (p < 0.001). Significant negative correlations (p < 0.001) were found between serum EPO levels and FEV1 (rs = -0.30), MEF25-75 (rs = -0.33), MEF50 (rs = -0.34). There was also a significant (p < 0.001) and negative correlation between ECP levels and FEV1 (rs = -0.31), MEF25-75 (rs = -0.31), MEF50 (rs = -0.32). A good positive correlation was found between peripheral eosinophil count and EPO levels (rs = 0.80, p < 0.001), and ECP levels (rs = 0.67, p < 0.001). We also found a significant positive correlation between clinical score and peripheral eosinophil count (rs = 0.54, p < 0.001), EPO levels (rs = 0.46, p < 0.001) and ECP levels (rs = 0.52, p < 0.001).     

Smoking and airway inflammation in patients with mild asthma.

STUDY OBJECTIVES: Cigarette smoking is common in asthmatic patients, and we investigated the impact of cigarette smoking on airway inflammation in asthma. DESIGN: Single-center observational study of airway inflammation in asthmatic and healthy smokers and nonsmokers. SETTING: Asthma research unit in a university hospital. PATIENTS OR PARTICIPANTS: Sixty-seven asthmatic and 30 nonasthmatic subjects classified as smokers or nonsmokers. Asthmatics had chronic, stable asthma and were not receiving inhaled or oral steroids at the time of the study. INTERVENTIONS: We examined induced-sputum cell counts and levels of interleukin (IL)-8 and eosinophilic cationic protein (ECP). Bronchial hyperreactivity was assessed using methacholine challenge. MEASUREMENTS AND RESULTS: Asthmatic smokers had higher total sputum cell counts than nonsmoking asthmatics and both smoking and nonsmoking healthy subjects. Smoking was associated with sputum neutrophilia in both asthmatics and nonasthmatics (median, 47% and 41%, respectively) compared with nonsmokers (median, 23% and 22%, respectively), and sputum IL-8 was increased in smokers compared with nonsmokers, both in subjects with asthma (median, 945 pg/mL vs 660 pg/mL, respectively) and in healthy subjects (median, 1,310 pg/mL vs 561 pg/mL, respectively). Sputum eosinophils and ECP levels were higher in both nonsmoking and smoking asthmatics than in healthy nonsmokers. In smoking asthmatics, lung function (FEV(1) percent predicted) was negatively related to both sputum IL-8 (r = - 0.52) and sputum neutrophil proportion (r = - 0.38), and sputum IL-8 correlated positively with smoking pack-years (r = 0.57) and percent neutrophil count (r = 0.51). CONCLUSIONS: In addition to the eosinophilic airway inflammation observed in patients with asthma, smoking induces neutrophilic airway inflammation; a relationship is apparent between smoking history, airway inflammation, and lung function in smoking asthmatics.     

Sputum eosinophilia in Churg-Strauss syndrome

Sputum induction (IS) can be used to study airway inflammation in asthmatics and other lung diseases. However, no data are available for patients with Churg-Strauss syndrome (CSS). A study was carried out to evaluate eosinophil counts and eosinophil cationic protein (ECP) levels in induced sputum during the follow-up of three patients with CSS. Induced sputum was carried out in 10 patients with corticosteroid-dependent asthma (used as a control group). Patients with CSS had significantly higher eosinophils percentages and ECP levels in sputum than those with stable corticosteroid-dependent asthma. During the follow-up, patients with CSS presented increased ECP levels sputum and eosinophils in sputum as well as increased blood eosinophils, despite their oral corticosteroid and immunosuppressive treatment. Eosinophil percentage in sputum and the total number of eosinophils in peripheral blood were more predictive of exacerbations of CSS than sputum ECP.