诱导支气管哮喘患者痰中IL-25、TSLP指标改变的临床价值

目的 研究诱导支气管哮喘（简称哮喘）患者痰中IL-25、胸腺基质淋巴细胞生成素（thymic stromal lymphopoietin,TSLP）指标改变的临床价值.方法 选取2012年10月至2013年12月泰山医学院附属医院就诊的哮喘患者52例作为哮喘组,另取52例健康查体者为正常对照组.按照常规方法收集诱导痰标本,应用酶联免疫吸附法（ELISA）对两组痰中IL-25以及TSLP水平进行检测,对比两组的水平差异.同时对于哮喘组患者均行抗炎药物治疗15d左右之后,检测外周血单核细胞（PBMCs）中的CD40、CD80、CD86表达水平,进行治疗前后两组表达水平.结果 哮喘组患者诱导痰中TSLP以及IL-25水平分别为（2.49±0.46） μg/L,（294.38±79.84） ng/L,均显著高于对照组的（1.45±0.36） μg/L和（181.38土56.73）ng/L,差异均有统计学意义（P＜0.05）.哮喘患者治疗后PBMCs中CD40、CD80、CD86的表达分别为（18.9±3.6）,（30.9±2.9）及（38.7±4.5）,均显著低于治疗前的（29.1±4.7）,（30.9±2.9）及（38.7±4.5）,差异均有统计学意义（P＜0.05）.结论 诱导哮喘患者痰中IL-25、TSLP指标改变的临床价值极高,为今后的哮喘治疗提供了新途径.     

诱导痰过敏毒素C5a在支气管哮喘中的变化特点研究

目的探讨诱导痰过敏毒素C5a在支气管哮喘中的变化特点及其意义。方法选取2015年1月-12月本院呼吸内科确诊的100例支气管哮喘患者作为研究对象,其中急性发作期患者59例(急性发作期组)、缓解期患者41例(缓解期组),另选取非哮喘自愿者50例作为对照组,检测三组诱导痰中过敏毒素C5a、血清中白细胞介素-17(IL-17)、白细胞介素-18(IL-18)、白细胞介素-23受体(IL-23r)的水平,测定三组对象的肺功能指标。结果三组对象诱导痰中过敏毒素C5a、IL-17、IL-18及IL-23r水平比较,急性发作期组、缓解期组和对照组的诱导痰中过敏毒素C5a、IL-17、IL-18及IL-23r水平逐渐降低且组间比较差异均具有统计学意义(P0.05);三组对象FEV_1/FVC、FEV_1%pred比较,急性发作期组、缓解期组和对照组的FEV_1/FVC、FEV_1%pred水平逐渐升高,且组间比较差异均具有统计学意义(P0.05);哮喘患者诱导痰中IL-17、IL-18及IL-23r水平与过敏毒素C5a呈显著的正相关关系(P0.05),FEV1%pred与过敏毒素C5a呈显著的负相关关系(P0.05)。结论过敏毒素C5a与支气管哮喘的发病有关,并且与患者的病情、气道炎症反应有关。     

我国西部地区支气管哮喘儿童血清Periostin、TSLP检测的临床意义

目的探究血清Periostin、TSLP,在我国西部地区儿童支气管哮喘发病过程中可能的作用及其临床意义。方法选择2017年1月-2019年10月我院收治的确诊支气管哮喘急性发作期患儿60例(发作组)、选择同期哮喘缓解期患儿60例(缓解组)、健康儿童60例(对照组),通过呼出气一氧化氮(fractional exhaled nitric oxide,FeNO)测定仪检测FeNO,采用ELISA法检测三组血清中Periostin、TSLP和IgE的表达水平,分析急性发作期不同病情的患儿血清Periostin、TSLP和IgE的水平,通过Pearson相关系数分析急性发作期患儿血清Periostin、TSLP和FeNO、IgE的相关性。结果发作组和缓解组患儿FeNO水平均显著高于对照组(P<0.05),且发作组患儿FeNO水平显著高于缓解组(P<0.05);发作组和缓解组患儿血清Periostin、TSLP和IgE表达水平均显著高于对照组(P<0.05),且发作组患儿血清Periostin、TSLP和IgE表达水平显著高于缓解组(P<0.05);重度组、中度组急性发作期组患儿血清Periostin、TSLP和IgE水平显著的高于轻度组患儿(P<0.05),重度组患儿的血清Periostin、TSLP和IgE水平显著的高于中度组(P<0.05);急性发作期哮喘患儿血清Periostin、TSLP与FeNO、IgE均呈显著正相关(P<0.05)。结论血清Periostin和TSLP可能在我国西部地区儿童支气管哮喘发病过程中扮演重要角色,并且与病情和发作情况相关,可作为潜在的诊断标志物及治疗靶点。     

支气管哮喘患者痰中细胞因子水平的变化

目的 研究细胞因子在哮喘发病机制中的作用,并为无创伤性监测哮喘气道炎症提供一种实用、客观的方法.方法 应用ELISA法检测42例哮喘患者及20例正常人痰中IL-6、IL-8、TNF-α水平.结果 缓解期哮喘患者诱导痰中IL-6、IL-8、TNF-α水平较正常对照组明显升高（分别为P〈0.05、P〈0.05和P〈0.002）,急性发作期哮喘患者治疗后痰中IL-6、IL-8水平较治疗前显著下降（分别为P〈0.05和P〈0.01）,而TNF-α则无统计学意义（P〉0.05）.结论 IL-6、IL-8、TNF-α积极参与了哮喘的发病,痰中IL-6、IL-8水平或可作为评价哮喘临床疗效的一项辅助指标,痰液分析在哮喘气道炎症研究中是一种有效的方法.     

哮喘患者血清TNF-α、IL-13、IgE水平变化及意义

采用ELISA法检测60例支气管哮喘患者观察组（急性发作期31例,缓解期29例）和22例健康体检者（对照组）血清IL-13、TNF-α、IgE水平。结果观察组TNF-α、IL-13、IgE水平均显著高于对照组（P均〈0.05）,发作期IL-13水平显著高于缓解期（P〈0.05）,发作期与缓解期TNF-α水平无明显差异。提示TNF-α、IL-13升高是哮喘发病的重要因素,IL-13促进IgE的产生。     

支气管哮喘患儿血清细胞因子水平变化及意义

目的探讨细胞因子在支气管哮喘发生、发展中的作用。方法采用双抗体夹心ELISA法检测100例支气管哮喘患儿（观察组,其中急性发作期及缓解期各50例）及30例健康儿童（对照组）血清IL-2、IL-6、IL-8、IL-10水平。结果观察组急性发作期患儿IL-6、IL-8水平明显高于缓解期及对照组,缓解期患儿IL-6、IL-8水平明显高于对照组,P均〈0.05;急性发作期患儿IL-2、IL-10水平明显低于缓解期及对照组,缓解期患儿IL-6、IL-8水平明显低于对照组,P均〈0.05。结论哮喘气道炎症的发生与IL-6、IL-8上调,IL-2、IL-10消耗性降低有关。血清IL-2、IL-6、IL-8、IL-10水平变化可用于判断支气管哮喘的病情并指导治疗。     

哮喘患儿血清IL-10、IL-13及IgE的测定及意义

采用ELISA法测定52例支气管哮喘患儿（观察组,其中急性发作期27例,缓解期25例）血清白细胞介素（IL）-10、TL-13和IgE水平,并与体检正常20例儿童（对照组）进行对照;对哮喘发作期IL-10、IL-13与IgE水平行相关分析。结果 急性发作期患儿血清IL-10水平显著低于缓解期和对照组（P均〈0.05）,而IL-13和IgE水平显著高于缓解期和对照组（P均〈0.05）。IL-10与IgE呈负相关（r=-0.568,P〈0.05）,IL-13与IgE呈正相关（r=0.593,P〈0.05）。认为IL-10和IL-13参与支气管哮喘发病的整个过程;检测血清IL-10、IL-13及IgE水平可反映支气管哮喘的炎症状态及评价治疗效果。     

急性发作期老年晚发哮喘患者气道炎症特征分析

目的检测急性发作期老年晚发哮喘(LOA)诱导痰细胞学、嗜酸细胞阳离子蛋白(ECP)和白介素-5(IL-5)、白介素-8(IL-8)水平,观察LOA气道炎症特征。方法检测86例急性发作期LOA患者诱导痰中细胞学分类计数、ECP和IL-5、IL-8水平;选择30例健康老年人作为对照。结果以诱导痰中EOS数量≥3%作为临界值,86例急性发作期LOA患者中,79例(81%)诱导痰中嗜酸细胞(EOS)数量增高,为痰EOS增高组;17例(19%)痰中性细胞数量增高,为痰非EOS增高组。痰非EOS增高组诱导痰中性细胞和IL-8水平显著高于痰EOS增高组和健康老年组(P0.01);而痰中EOS数量、ECP和IL-5水平显著低于痰EOS增高组(P0.01),但与健康老年组比较差异无统计学意义(P0.05)。结论急性发作期LOA患者存在气道嗜酸细胞和中性细胞两种炎症类型,测定患者气道炎症类型有助于指导治疗。     

白细胞介素13在哮喘、COPD中的变化研究

目的探讨白细胞介素13（IL-13）在支气管哮喘、慢性阻塞性肺疾病（COPD）的变化。方法选择正常组30例、哮喘急性发作期和COPD急性加重期各30例，哮喘缓解期和COPD稳定期各30例。取静脉血，用ELISA方法测定血清中IL-13的水平，并进行统计学分析。结果哮喘IL-13水平分析：急性发作期68．18±2．24（pg／ml）显著高于缓解期43．49±4．04（pg／ml），P〈0．01，且两组均显著高于正常对照组25．45±5．49（pg／m1），P〈0．01；COPD IL-13水平分析：急性加重期组51．55±3．16（pg／m1）显著高于稳定期组34．89±2．16（pg／m1），P〈0．01。且两组均显著高于对照组，P〈0．01。结论IL-13参与了哮喘和COPD慢性炎症的形成。     

哮喘患者诱导痰IL-8、MMP-9水平检测及其临床意义

目的探讨哮喘患者发病及治疗后诱导痰中炎症因子的变化及意义。方法随机选取哮喘患者135例、慢性阻塞性肺疾病（COPD）患者20例、健康对照者20例,治疗前后行诱导痰炎性细胞分类并计数,检测痰上清液IL-8、基质金属蛋白酶9（MMP-9）、嗜酸性粒细胞趋化因子Eotaxin-1水平。结果哮喘患者诱导痰中嗜酸性粒细胞比例及Eotaxin-1水平高于对照组及COPD组,中性粒细胞比例及IL-8、MMP-9水平高于对照组但低于COPD组,且重度哮喘患者上述指标均高于轻中度哮喘患者;哮喘患者中性粒细胞比例及IL-8、MMP-9水平均随治疗后好转有所降低。结论中性粒细胞及炎症介质IL-8与MMP-9对于评价哮喘患者病情及控制水平有一定意义。     

Correlation of TSLP,IL-33, and CD4 + CD25 + FOXP3 + T regulatory (Treg) in pediatric asthma

Objective: Newly discovered cytokines TSLP and IL33 are being studied as important indicators of Th2 inflammation and their effect on Treg cells is likely to modulate immune response. We attempted to study TSLP and IL-33 and then correlated with Tregs in order to find possible biomarker in these patients. Methods: Sixty-five children (37 with asthma only and 28 with asthma and rhinitis) aged 6.4?±?3.2 years (patient group) and 15 healthy children aged 8.0?±?2.6 years (control group) were recruited in this study. In vitro analysis of TSLP and IL-33 was done in serum samples of 65 newly diagnosed children for allergic asthma and 15 healthy children using the sandwich ELISA method. The expression of Treg cells (CD4?+?CD25?+?FOXP3+) was analyzed by flow cytometry. Results: The mean TSLP in the patient group (592?±?68?pg/ml) was significantly higher than controls (215?±?45?pg/ml) (p?<?0.05). Alternatively, the expression of FOXP3?+?T reg cells was significantly lower in the patient group (52?±?36) compared with the controls (95.9?±?3.6) (p?=?0.003). IL-33 was also significantly higher (4044?±?413?pg/ml) in the patient group compared with the controls (3282?±?331.5?pg/ml) (p?=?0.0001). The expression of Treg cells was negatively correlated with the TSLP (r?=??0.23, p?=?0.07). Asthma control test (ACT) was also negatively correlated with TSLP in the patient group (r?=??0.14, p?>?0.05). Conclusion: Children with asthma show elevated serum levels of TSLP, which correlated negatively with asthma control test and Treg cells. TSLP may be used as a biomarker for inflammation in pediatric asthma patients.     

南蛇藤素对哮喘小鼠TSLP介导的Th2优势免疫干预的研究

目的观察南蛇藤素对小鼠哮喘模型胸腺基质淋巴生成素(TSLP)介导的气道炎症的影响及机制。方法 24只SPF级BALB/c雌性小鼠随机分成4组。哮喘组、甲强龙组及南蛇藤素组分别予OVA致敏,然后腹腔注射等体积药物(分别为生理盐水、甲强龙及南蛇藤素),再予以雾化吸入1%OVA激发。对照组使用等剂量生理盐水。末次激发24h后,检测各组小鼠血清总Ig E及TSLP含量,支气管肺泡灌洗液(BALF)中IL-4、IL-13水平,HE染色肺组织病理学检测炎症浸润。结果南蛇藤素组小鼠哮喘表现较哮喘组明显改善;其血清总Ig E、TSLP含量及BALF中IL-4、IL-13水平均低于哮喘组(P0.05);且小鼠肺组织气道炎症明显减轻,但与甲强龙组无异(P0.05)。结论南蛇藤素组可以通过抑制哮喘小鼠TSLP介导的Th2优势免疫改善其临床症状。     

氧化低密度脂蛋白诱导血管内皮细胞表达胸腺基质淋巴细胞生成素

目的:探讨氧化低密度脂蛋白(ox-LDL)是否可以诱导血管内皮细胞产生胸腺基质淋巴细胞生成素(TSLP)。方法:将25mg/L及50mg/L的ox-LDL与原代培养的人脐静脉内皮细胞(HUVEC)共育6h,用RT-PCR检测HUVEC TSLP mRNA的表达、免疫组织化学染色检测HUVEC TSLP蛋白的表达,ELISA法检测培养上清中TSLP的浓度。结果:对照组HUVEC几乎不表达TSLP;ox-LDL刺激6h后,HUVEC可表达TSLPmRNA,胞浆中存在TSLP;上清中TSLP的浓度显著高于对照组,并且随ox-LDL刺激浓度的升高,TSLP的表达升高。结论:ox-LDL可诱导血管内皮细胞产生TSLP,这一作用可能在血管的炎症反应中起作用。     

Thymic stromal lymphopoietin contribution to the recruitment of circulating fibrocytes to the lung in a mouse model of chronic allergic asthma

Objective: Fibrocyte localization to the airways and thymic stromal lymphopoietin (TSLP) overexpression in the lung are features of severe asthma. The aim of this study was to determine whether TSLP contributes to fibrocyte trafficking and airway remodeling in a mouse model of allergic asthma. Methods: We established a chronic asthma animal model by administering house dust mite (HDM) extracts intranasally for up to 5 consecutive weeks. Mouse anti-TSLP monoclonal antibody (mAb) was given intraperitoneally starting the 4^th week. Fluorescence-labeled CD34/collagen I (Col I)-dual-positive fibrocytes were examined by confocal microscopy. The level of TGF-β1 in the bronchoalveolar lavage (BAL) fluid was determined by ELISA. Results: We found significantly increased levels of TSLP and TGF-β1 in the lung of the mice subjected to repeated allergen exposure, which was accompanied by increased number of fibrocytes in the sub-epithelial zone and the BAL fluid. However, blocking TSLP markedly decreased the production of TGF-β1, reduced the number of fibrocytes and subsequently prevented alterations of both airway and vascular structures. Conclusions: Our data suggested that TSLP might function in airway remodeling by promoting circulating fibrocyte recruitment to the lung in the mice subjected to chronic allergen exposure. These results provide a better rationale for targeting the interaction between TSLP and fibrocytes as a therapeutic approach for chronic allergic asthma.     

Anti-TSLP antibodies: Targeting a master regulator of type 2 immune responses

TSLP is an epithelial cell-derived cytokine synthesized in response to various stimuli, including protease allergens and microorganisms like viruses and bacteria. Biological functions of TSLP require heterodimer formation between the TSLP receptor (TSLPR) and IL-7 receptor-α, which polarize dendritic cells to induce type 2 inflammation and directly expand and/or activate Th2 cells, group 2 innate lymphoid cells, basophils, and other immune cells. TSLP is thus considered a master regulator of type 2 immune responses at the barrier surfaces of skin and the respiratory/gastrointestinal tract. Indeed, genetic, experimental, and clinical evidence suggests that the TSLP-TSLPR pathway is associated with the pathogenesis of allergic diseases such as atopic dermatitis (AD) and asthma. Tezepelumab (AMG-157/MEDI9929) is a human anti-TSLP antibody that prevents TSLP-TSLPR interactions. A phase 2 trial for moderate to severe AD showed that a greater but not statistically significant percentage of tezepelumab-treated patients showed clinical improvements compared to the placebo group. A phase 2 trial for uncontrolled, severe asthma showed significant decreases in asthma exacerbation rate and improved pulmonary function and asthma control for tezepelumab-treated patients. Levels of biomarkers of type 2 inflammation, such as blood/sputum eosinophil counts and fraction of exhaled nitric oxide decreased, however, clinical efficacy was observed irrespective of the baseline levels of these biomarkers. A blockade of the TSLP-TSLPR pathway likely will exert significant clinical effects on AD, asthma, and other allergic diseases. The efficacy of anti-TSLP antibodies compared to other biologics needs to be further examined.     

TSLP基因多态性与支气管哮喘遗传易感性的关系

支气管哮喘（简称哮喘）是多基因参与的具有遗传易感性的慢性气道变应性炎症。胸腺基质淋巴细胞生成素（TSLP）是一种由上皮细胞产生的白介素7样细胞因子,与靶细胞TSLP受体相互作用,激活骨髓源性树突状细胞,并诱导一种特殊类型的炎症性Th2免疫应答,参与哮喘的发生和发展。TSLP还可通过调节核因子κB,参与哮喘的发病过程。多数家系聚集、孪生和大的遗传队列研究均证明,哮喘存在遗传易感性。靶向TSLP有望为今后哮喘免疫治疗带来新的前景。     

胸腺基质淋巴细胞生成素与支气管哮喘

胸腺基质淋巴细胞生成素(TSLP)是一种由上皮细胞产生的白介素7样细胞因子,与靶细胞TSLP受体相互作用,可以激活骨髓源性树突状细胞,进而诱导一种特殊类型的炎症型Th2细胞反应,而与支气管哮喘(简称哮喘)相关的致病性T细胞正是这类炎症型T细胞.TSLP还可诱导肥大细胞分泌Th2细胞因子,参与哮喘的发病过程.