Effect of house dust mite immunotherapy on interleukin-10- secreting regulatory T cells in asthmatic children

Background Subcutaneous specific immunotherapy has been demonstrated to be capable of inducing T-cell regulatory response.Interleukin-10 （IL-10） plays a crucial role in inducing allergen-specific tolerance.However the reports of the changes of IL-10 in house dust mite （HDM）-specific immunotherapy were varied.The aim of this study was to evaluate the function of IL-10-secreting regulatory T cells in asthma children successfully treated with HDM immunotherapy.Methods Peripheral blood mononuclear cells （PBMCs） were isolated from 27 patients following 1.5--2 years of HDM-specific immunotherapy （SIT, SIT group） and from 27 matched treated asthmatic children allergic to HDM （asthmagroup）.After 48 hours of in vitro stimulation with HDM extracts, IL-10-secreting regulatory T cells were measured by four colour flow cytometry.Sera were tested for allergen-specific IgG4 and IgE using the Immuno CAP 100 assay.Results PBMCs from children undergoing immunotherapy following HDM extracts stimuli produced significantly more IL-10 compared with the asthma group.The frequency of iTreg cells and aTreg cells increased in SIT group after HDM stimulation, while it was not affected in the asthma group.Among the iTreg cells and aTreg cells, the frequency of CD4＋CD25-Foxp3-IL-10＋ Treg cells increased the most which was 2 times higher than that in unstimulated cultures in SIT group.The levels of HDM-specific IgG4 of SIT group was significiently higher compared with asthma group, but there was no correlation of the levels of HDM-specific IgG4 and IL-10 secreting Treg cells.Conclusions HDM-specific immunotherapy can successfully upregulate the frequency of IL-10-secreting Treg cells.CD4＋CD25-Foxp3-IL-10＋ Treg cells may play a key role in inducing the immune tolerance in HDM-specific immunotherapy.     

Effect of sublingual immunotherapy on level of cytokines in PBMCs of patients with allergic asthma

This study examined the possible mechanism of sublingual immunotherapy(SLIT) in the treatment of allergic asthma.Forty asthma patients allergic to dust mite were enrolled.They received SLIT with dermatophagoides farinae(Der.f) drops for one year.Thirty healthy subjects served as controls.The levels of IL-4 and IFN-γ of peripheral blood mononuclear cells(PBMCs) were determined in allergic asthma patients before and after the SLIT as well as the healthy subjects.The results showed that the level of IL-4 was substantially increased and that of IFN-γ remarkably decreased in the patients before the SLIT as compared with those in the healthy subjects(P<0.05).After the SLIT,the level of IL-4 was significantly reduced and that of IFN-γ elevated in these allergic asthma patients.It was concluded that sublingual immunotherapy is effective for patients with allergic asthma.And it may work by regulating the balance of Th1/Th2 through changing the expression of IL-4 and IFN-γ in PBMCs.     

The Effect of Interleukin-18 on Airway Inflammation in Asthmatic Murine Models and Its Mechanisms

In order to investigate the effect of interleukin-18 （IL-18） on airway inflammation in asthmatic murine models and its mechanisms, BALB/C mice were randomly divided into three groups （n=10 in each group）： group A （control group）; group B （asthmatic model group）; group C （IL-18-treated group）. The asthmatic model was established in groups B and C by respiratory syncytial virus （RSV） killed by ultraviolet. Saline solution （0.1 mL） and IL-18 （0.1 mL, 1μg） were intraperitoneally injected respectively in groups B and C at 7 time points （day 1, 2, 7, 8, 9, 21, 22）. The number of eosinophils （EOS） and plasmacytes in the airway was observed. The levels of interferon gamma （IFN-γ） in bronchoalveolar lavage fluid （BALF） were measured by ELISA. The results showed that symptoms of asthma in group C were more severe than in groups A and B. In group A, there were no EOS and plasmacytes in the airway submucosa. The number of EOS [ 15±3 （average cell counts per microscopic visual field, the same below）] and plasmacytes （10±2） in group B were increased significantly. However, the number of EOS and plasmacytes in group C （6±2 and 2±1, respectively） was decreased significantly as compared with group B （both P〈0.05）. The levels of IFN-γ in groups A, B and C were 31±3, 40±5 and 63±5 μg/mL respectively, and those in group C were significantly higher than in groups A and B （both p〈0.05）. It was suggested that the mechanism by which IL-18 inhibited the airway inflammation in asthmatic mice might be contributed to the fact that IL-18 could induce the induction of IFN-γ.     

Effect of ligustrazine injection on levels of interleukin-4 and interferon-gamma in patients with bronchial asthma

Objective： To explore the effect of ligustrazine injection （LI） on serum levels of interleukin-4 （IL-4） and interferon- -γ （IFN- -γ ） in patients with bronchial asthma and determine the mechanism of action of LI in preventing and treating asthma. Methods： Sixty-eight patients with mild or moderate bronchial asthma were assigned to two groups equally according to their sequence number, odd or even. The conventional treatment was administered to both groups, and LI was given to the treatment group by ultrasonic spray inhalation twice a day but not to the control group. The therapeutic course for all was 2 weeks. Further, 30 healthy subjects who had no history of smoking were enrolled as the normal control group. The clinical condition scores, frequency of attacks and dosage of Terbutaline inhaled were scored and recorded on the first day of hospitalization （before treatment） and after treatment. At the same time, the indexes of lung function, including forced expiratory volume in one second （FEV1）, forced expiratory volume in one second to forced vital capacity ratio （FEV1%） and the peak expiratory flow （PEF） were determined before treatment. The levels of IL-4 and IFN--γ in peripheral blood were detected by ELISA before and after treatment, and compared with that of the healthy control group. Results： After treatment, the clinical condition scores were found to be lower, indexes of lung function were elevated, but serum level of IL-4 and ratio of IL-4/IFN-γwere reduced in both groups, showing significant differences as compared to those before treatment （P〈0.05）. However, the changes in all the indexes were more significant in the treatment group than in the control group, also showing statistical significance （P〈0.05）. No significant difference was revealed when IFN--γ levels were compared before and after treatment in both groups, though a lowering trend could be seen, significant difference could not be found in the comparison of IFN--γ levels between groups a     

Effect of interleukin-33 on Th1/Th2 cytokine ratio in peripheral lymphocytes in asthmatic mice

Background Allergic asthma is a chronic airway inflammatory disease partly characterised by high concentration of T help 2 （Th2） cytokines in bronchoalveolar lavage fluid （BALF）. There is no report on the relation of peripherally circulating blood lymphocytes and asthma. We explored the balance of Th2fThl cytokines in asthmatic mice. Exogenous recombinant interleukin （IL） 33 acted on murine peripheral circulating blood lymphocytes, IL-5 cytokine was selected for assessing Th2 cytokines and interferon-gamma （IFN-~） for Thl cytokines. Methods Female specific pathogen free BABL/c mice were sensitised by intraperitoneal injection of 20 IJg of ovalbumin emulsified in 1 mg of aluminium hydroxide gel in a total volume of 200 IJI, and challenged for 30 minutes in 7 consecutive days with an aerosol of 2 g ovalbumin in 100 ml of PBS. Then we collected BALF and isolated lymphocytes from the peripheral blood. The lymphocytes were divided into two groups： asthmatic group and normal group. Thl/Th2 cytokines was detected by enzyme-linked immunosorbent assay （ELISA） kits. Results In the asthma group, we found numerous eosinophils and lymphocytes on the glass slides. We then confirmed that the optimal concentration of IL-33 was 10 ng/ml and time of IL-33 stimulating lymphocytes was 24 hours. In the asthma group, the production of IL-5 was significantly increased over normal group after stimulation with IL-33 （P 〈0.05） and the production of IFNy was supressed from IL-33 stimulated lymphocytes （P 〈0.05）. Conclusion IL-33 acts on lymphocytes of peripheral blood increasing secretion of Th2 cytokines and inhibiting secretion of Thl cytokines.     

The Effect of Ginkgo Biloba Extract on the Expression of PKCα in the Inflammatory Cells and the Level of IL-5 in Induced Sputum of Asthmatic Patients

To investigate the effect of the Ginkgo Biloba Extract （GBE） on the asthma and examine its possible mechanisms, 75 asthma patients were divided into 4 groups and the patients were respectively treated with fluticasone propionate for 2 weeks or 4 weeks, or treated with fluticasone propionate plus GBE for 2 weeks or 4 weeks. Fifteen healthy volunteers served as healthy controls. Sputum inhalation with inhaling hypertonic saline （4%-5%） was performed. Lung ventilatory function and forced expiratory volume in one second （FEVI） were measured. The numbers of different cells in induced sputum were calculated. The expression of PKCα in the cells was immunocytochemically detected and the percentages of positive cells in different cells were counted. Interleukin-5 （IL-5） in sputum supernatants was detected with enzyme-linked immunosorbent assay. The percentage of eosinophils, lymphocytes, PKCα positive inflammatory cells and the concentration of IL-5 in asthmatic patients were higher than those in the controls （P〈0.05）, and the eosinophils, lymphocytes, positive expression of PKCα and the level of IL-5 were significantly decreased in asthmatic patients after they were treated with fluticasone propionate or fluticasone propionate plus GBE. However, they were still significantly higher than those of the controls. Compared to the group treated with glucocorticosteroid for 2 weeks, no significant decrease was found in the percentage of eosinophils, lymphocytes, PKCα positive inflammatory cells and the IL-5 in the supernatant of induced sputum. Compared with the group treated with glucocorticosteroid for 2 or 4 weeks, significant decrease in the same parameters was observed in the group treated with fluticasone propionate and GBE for 4 weeks. The IL-5 level in the supernatant of induced sputum was positively correlated with the percentage of PKCα-positive inflammatory cells and the percentage of eosinophils in the induced sputum in asthma patient groups respectively （n=150, r=0.83, P〈0.01; n=150,     

Changes of sulfur dioxide, nuclear factor-κB, and interleukin-8 levels in pediatric acute lymphoblastic leukemia with bacterial inflammation

Background Bacterial inflammation is a common complication in patients with leukemia,and sulfur dioxide （SO2） is a bioactive molecule in modulating Gram-negative bacilli infection.This study aimed to examine the changes in SO2,nuclear factor-κB （NF-κB）,and interleukin-8 （IL-8） levels in pediatric acute lymphoblastic leukemia （ALL） with Gram-negative bacterial inflammation.Methods Fifty-five ALL children were enrolled in this study,including 30 males and 25 females,aged 3-13 years,and the median age was 7.8 years.All these children who accepted chemotherapy for ALL were divided into the control group （before chemotherapy）,the infection group （after chemotherapy with infection）,and the recovery group （the infection was controlled after 1 week）.The serum level of SO2 was detected using high performance liquid chromatography with fluorescence assay,and NF-κB and IL-8 levels were measured by enzyme-linked immunosorbent assay （ELISA）.Human THP-1 cells were cultured,induced,and differentiated into macrophages,which were divided into five groups and each group was cultured with different stimulators：lipopolysaccharide （LPS） group,LPS＋L-aspartate-β-hydroxamate （HDX） group,LPS＋SO2 group,SO2,and control groups.NF-κB level and IL-8 protein contents by ELISA were examined in each group.Results In comparison with those of the control group,levels of serum SO2,NF-κB,and IL-8 of the infection group were significantly increased （P ＜0.05）,while those of the recovery group were significantly decreased （P ＜0.05）.A positive correlation was found between levels of serum SO2 and intracellular NF-κB/IL-8,and the correlation coefficients were 0.671 and 0.798 （P ＜0.05）,respectively.According to the results found in human THP-1 cells,levels of NF-κB and IL-8 in LPS group were significantly increased compared with those of the control group （P ＜0.05）; when compared with those in LPS group,levels of NF-κB in LPS＋HDX group further increased significantly （P ＜     

Pulmonary artery perfusion with HTK solution prevents lung injury in infants after cardiopulmonary bypass

Background Pulmonary artery perfusion during cardiopulmonary bypass （CPB） is a novel adjunctive method, which can minimize the lung ischemic-reperfusion injury and inflammatory response. This study evaluated the protective effect of pulmonary perfusion with hypothermic HTK solution in corrections of congenital heart defects with pulmonary hypertension. Methods Between June 2009 and December 2009, 24 consecutive infants with congenital heart defects and pulmonary hypertension were randomly divided into perfused group （n=12） and control group （n=-12）. Oxygen index, alveolar-arterial 02 gradient, serum levels of malondialchehyche （MDA）, interleukin （IL）-6, -8, -10, soluble intercellular adhesion molecule-1 （slCAM-1）, and P-seiectin were measured before commencement and serially for 48 hours after termination of bypass. Results Oxygenation values were better preserved in the perfused group than in the control group. The serum levels of IL-6 increased immediately after CPB in both groups and returned to baseline at 48 hours after CPB, but it was restored faster and earlier in the perfused group. The serum levels of IL-8, slCAMol, and MDA remained at baseline at each point after CPB in the perfused group and elevated significantly immediately after CPB in the control group, except for slCAM-1 The serum level of IL-10 increased immediately after CPB and decreased to baseline at 48 hours after CPB in both groups, but the IL-10 level in the perfused group was significantly higher than in the control group at 12 hours after CPB. The serum P-selectin levels in the control group immediately after CPB were significantly higher than prebypass levels. Moreover, there were no significant differences in postoperative clinical characters, except for the intubated time. Conclusion In infants with congenital heart defects, pulmonary perfusion with hypothermic HTK solution during cardiopulmonary bypass could ameliorate lung function and reduce the inflammatory response.     

屋尘螨变应原特异性免疫治疗对变应性哮喘患儿预后的影响

目的 探讨屋尘螨变应原特异性免疫治疗(SIT)对变应性哮喘患儿预后的影响.方法 选择尘螨变应原皮试阳性的过敏性支气管哮喘患者42例,进行SIT治疗1年,于屋尘螨变应原疫苗皮下注射治疗前及疗程结束后分别取外周血,测定外周血清IL-4、IFN-γ水平及检测肺功能.选择同期在我院哮喘专科门诊就诊,确诊为儿童哮喘、尘螨变应原皮试阳性患儿23例作为对照组.结果 治疗1年后,SIT组儿童血清IL-4水平下降、IFN-γ水平及IFN-γ/IL-4升高(P<0.05),肺功能指标FVC、pre-FEV1%、pre-PEF%改善明显(P=0.05);而对照组儿童IFN-γ水平及IFN-γ/IL-4升高(P<0.05),血清IL-4水平较治疗前无明显下降(P<0.05),肺功能指标FVC、pre-FEVI%、pre.PEF%改善不明显(P>0.05);组间比较,两组患儿治疗前肺功能指标FVC、pre-FEVI%、pre-PEF%差异无显著性,治疗1年后,SIT组pro-FEVI%、pre-PEF%显著优于对照组.结论 SIT能够调节体内Th1/Th2细胞的平衡,维持哮喘患儿良好的肺功能,SIT可能对哮喘儿童气道重塑有一定抑制作用.     

长期舌下含服粉尘螨滴剂治疗儿童支气管哮喘的有效性和安全性评价

目的：观察长期舌下含服标准化粉尘螨滴剂治疗儿童支气管哮喘的有效性和安全
性,为儿童支气管哮喘的治疗提供一种安全、有效的方法。方法：采用随机对照研究方法,
将本院哮喘专科诊治的对粉尘螨过敏的4～14岁轻-中度哮喘患儿分为治疗组（64例）和对
照组（64例）。对照组患儿仅接受对症药物治疗,治疗组患儿在此基础上联合粉尘螨滴剂舌
下含服进行特异性免疫治疗,在治疗后的6个月、1年及2年进行随访评估。对2组患儿治疗前、后的哮喘
症状进行评分,并对呼气峰流速值占正常预计值的百分比(PEF％)进行统计学分析比较。结果：在128例研究对象中,治疗组有5例病例脱失。治疗组患儿在治疗第6个月、1年和2年的哮喘症状评分较对照组各阶段症状评分均有所下降,差异均具有统计学意义（P<0.05）。治疗组患儿在治疗第6个月、1年、2年时间段的PEF％较对照组各阶段均明显升高(P<0.05)。免疫治疗中少数患儿出现过轻微反应,均未出现严重的不良反应。结论：长期舌下含服粉尘螨滴剂能明显改善哮喘患儿的症状及肺功能,不良反应发生率低且轻微,是一种治疗儿童支气管哮喘安全、有效的方法。     

甘草酸二铵辅助治疗咳嗽变异性哮喘发作期的临床研究

目的观察甘草酸二铵辅助治疗对咳嗽变异性哮喘（CVA）患者血中IL-4、IL-5和IL-13水平的影响。方法将100例CVA患者随机平均分为吸入激素组和甘草酸二铵＋吸入激素组。分别在治疗前和治疗4周后对患者进行肺功能、PEF％和激发试验检查和咳嗽评分,并对患者血中IL-4、IL-5和IL-13水平进行测定。结果两组患者在治疗前后肺功能指标FEV1％占预计值和FEV1／FVC％及PEF％未见明显区别。但两组患者治疗后PC20明显升高,且甘草酸二铵＋吸入激素组患者PC20升高更加明显。治疗4周后患者咳嗽评分及血中IL-4、IL-5和IL-13水平较治疗前明显下降,甘草酸二铵＋吸入激素组患者下降更加明显。结论甘草酸二铵可以有效的减轻CVA患者的临床症状和AHR,且同时还可以抑制CVA导致的气道炎症反应。在常规吸入激素的基础上使用甘草酸二铵辅助治疗是一种安全而有效的联合治疗方案,值得临床推广应用。     

标准化免疫治疗对哮喘儿童屋尘螨特异性IgG4和肺功能的影响

目的 探讨标准化屋尘螨变应原制剂免疫治疗儿童哮喘,对其肺功能、气道反应性及血清屋尘螨特异性IgG4、血清屋尘螨特异性IgE的影响.方法 选择31例轻-中度屋尘螨过敏性哮喘儿童,给予标准化屋尘螨变应原制剂治疗,疗程25周,检测治疗前后肺功能指标FEV1和PEF占预计值的百分比,PC20、血清屋尘螨特异性IgG4和屋尘螨特...     

舌下特异性免疫治疗儿童过敏性哮喘86例临床疗效观察

目的：研究粉尘螨滴剂舌下特异性免疫（SLIT）治疗儿童哮喘的有效性。方法：将确诊为变态反应性哮喘且粉尘螨变应原皮肤点刺试验呈阳性反应86例患儿随机分为SLIT组（n=45）和常规治疗对照组（n=41）。常规治疗组按照儿童哮喘规范化治疗方案治疗,SLIT组在常规规范化治疗的基础上,加用舌下含服粉尘螨滴剂,观察两组治疗后哮喘临床症状改善情况。并分别于治疗前、后抽取患儿静脉血,分离血清,采用ELISA法测定白细胞介素-4（IL-4）及7干扰素（IFN-γ）的血清含量。结果：经治疗24周后,SLIT组的症状评分降低值及临床有效率均明显高于常规治疗组,差异均有统计学意义（P〈0．05）;SLIT组部分患儿出现红肿瘙痒等局部反应,均于次日自行消失。与治疗前相比,对照组治疗后IL-4及IFN-γ血清含量比较,差异无统计学意义（P〉0．05）;而SLIT组经过SLIT治疗后,IL-4水平明显下降,IFN-γ水平明显上升,差异具有统计学意义（P〈0．05）;治疗后SLIT组较对照组IL-4水平下降,IFN—γ水平上升,差异均有统计学意义（P〈0．05）。结论：粉尘螨滴剂舌下免疫能改善哮喘儿童的症状,纠正Th1／Th2的平衡,对儿童变态反应性哮喘具有良好的效果。     

520例儿童支气管哮喘过敏原点刺试验临床分析

目的:通过点刺液过敏原筛查引起本地区支气管哮喘的过敏原状况,从而达到有效的防控.方法:采用阿罗格标准化生产的点刺液筛查支气管哮喘过敏原,对520例哮喘儿童和300例健康儿童进行诊断.结果:520例患者中324例(62.7%)呈阳性.324例皮肤点刺试验中屋尘螨阳性率最高,其次是粉尘螨等.屋尘螨和粉尘螨支气管哮喘患儿与对...     

尘螨特异性免疫治疗对哮喘患儿的干预及对肺功能的影响

目的探讨尘螨特异性免疫治疗(SIT)与哮喘患儿IgE的相关性及对肺功能的影响。方法将56例哮喘轻-中度发作期儿童,随机分为A组35例(加用SIT治疗),B组21例(对照组)。在SIT前和SIT后6个月检测过敏原皮试、血浆IgE、肺功能,并进行临床疗效评价。结果 A组SIT后血浆总IgE、尘螨特异性IgE下降,过敏原复查有意义;临床疗效比较,A组有效率93.86%,B组为82%;两组肺通气功能均明显改善。结论 SIT可减少IgE的合成;SIT可明显改善肺通气功能,结合尘螨特异性IgE下降、临床疗效好,为哮喘用药及调整提供客观依据。     

过敏原检测对儿童哮喘的临床意义

目的探讨皮肤过敏原点刺试验、血清过敏原检测对儿童哮喘诊断、治疗的临床意义。方法应用粉尘螨及屋尘螨过敏原对120例哮喘患儿进行皮肤点刺试验,其中61例哮喘患者应用进行血清过敏原检测。结果（I）120例哮喘患者尘螨过敏原皮肤点刺试验总阳性率为33．33％,高于对照组（P〈0．05）。（2）粉尘螨过敏原阳性35例（29．17％）;屋尘螨过敏原阳性3I例（25．81％）。（3）粉尘螨及屋尘螨过敏原皮肤点刺试验结果与哮喘急性发作严重程度有关。（4）61例哮喘患者血清过敏原检测阳性48例（78．69％）,其中狗毛11例（18．03％）,猫毛10例（16．40％）,粉尘螨17例（27．87％）,屋尘螨15例（24．59％）,海鲜组合12例（19．67％）。（5）皮肤点刺试验与血清过敏原检测的尘螨过敏原检出率,差异无统计学意义（P〉0．05）。结论皮肤点刺试验可作为检测儿童哮喘过敏原的方法之一。     

儿童支气管哮喘常见变应原调查

①目的了解青岛地区支气管哮喘儿童常见吸入性和食物性变应原的情况。②方法采用皮肤点刺法，对1454例支气管哮喘儿童进行变应原检测。其中吸入性变应原16种，食物性变应原8种。③结果吸入性变应原中粉尘螨和屋尘螨的阳性与强阳性反应率较高，分别为57．1％和62．1％，其中有23．2％的病儿仅表现为粉尘螨和屋尘螨的强阳性反应。食物性变应原中以海虾、蚌类、花生的阳性与强阳性反应率较高，分别为15．2％、24．1％和16．4％。④结论青岛地区诱发儿童支气管哮喘的主要吸入性变应原为粉尘螨和屋尘螨，食物性变应原主要为海虾、蚌类、花生。     

阿奇霉素对哮喘患者诱导痰中细胞因子的影响

 目的 评价阿奇霉素对支气管哮喘患者诱导痰中细胞因子的影响及其安全性。方法 本实验40例哮喘患者,随机分为阿奇霉素联合吸入性糖皮质激素（inhaled cortisteroid,ICS）加长效β₂-肾上腺素受体激动剂（long-acting β₂-agonists,LABA）组（A组）和ICS加LABA组（B组）,治疗12周后,用ELISA检测患者治疗前后诱导痰中细胞因子IL-4、IL-5、IL-8、IL-10、IL-13、TNF-α、IFN-γ的浓度变化,并评估阿奇霉素治疗的不良反应。结果 治疗12周后,两组患者的哮喘诱导痰细胞因子的浓度明显下降,差异有统计学意义;A组诱导痰标本内细胞因子浓度较B组明显下降,差异有统计学意义;两组均未出现严重的不良事件。结论 阿奇霉素联合ICS和LABA治疗哮喘安全有效,可有效降低气道炎症反应程度,调节Th1/Th2平衡。