葎草花粉变应原耐受性树突状细胞对小鼠气道变应性炎症抑制作用的研究  

Inhibitory effect of Humulus scandens pollen allergen tolerated dendritic cells on mouse allergic airwayinflammation

作　　者：杨侠[1] 武润苗 张明[1] 单虎[1] 张秋红[1] 张洁[1] 李雅莉[1] 

机构地区：[1]西安交通大学第二附属医院呼吸内科,710004 [2]陕西省人民医院呼吸内科,西安710068

出　　处：《国际呼吸杂志》2017年第21期1623-1629,共7页International Journal of Respiration

基　　金：陕西省科技攻关项目（2014k11-02-03-08、2011k12-01-03）

摘　　要：目的用葎草花粉刺激的未成熟树突状细胞免疫小鼠，观察其对葎草花粉诱导哮喘小鼠气道炎症的抑制作用。方法采用流式细胞术分析葎草花粉变应原刺激培养出的未成熟树突状细胞表型变化，通过混合淋巴细胞反应检测其诱导同种异体T细胞的应答能力。BALB／c小鼠随机分为4组，哮喘组、葎草花粉变应原特异性未成熟树突状细胞组、地塞米松治疗组、正常对照组。应用葎草花粉变应原诱导出哮喘模型。激发后观察小鼠症状，肺组织行HE染色观察肺部炎性细胞浸润、水肿及气道上皮损伤情况；计算BALF细胞总数，涂片行Wright—Gimsa染色，计算出嗜酸粒细胞的百分比；培养脾细胞，流式细胞仪检测脾脏CD4-CD25＋Foxp3＋调节性T细胞百分比；酶联免疫吸附测定检测血清IgE、BALF及脾细胞上清液Th2型细胞因子IL-4及Th1型细胞因子干扰素-7的含量。结果蓓草花粉刺激的未成熟树突状细胞免疫哮喘小鼠后，能够有效抑制嗜酸粒细胞浸润和杯状细胞增生、分泌，减轻上皮细胞损伤，降低肺泡灌洗液中细胞总数、嗜酸粒细胞百分比、血清中IgE含量、BALF中Th2型细胞因子IL-4含量、脾细胞上清液中IL-4含量，提高脾脏CD4-CD25＋Foxp3＋调节性T细胞的百分比。结论葎草花粉刺激的未成熟树突状细胞疫苗对律草花粉诱导的哮喘小鼠气道炎症具有一定的抑制作用。Objective Preparation of Humulus pollen allergen tolerated dendritic cells,observe The inhibitory effect of Humulus scandens pollen allergen tolerated dendritic cells on mouse allergic airway inflammation. Methods Humulus pollen allergen tolerated dendritic cells were detected by flow cytometry and its capacity of inducing allogeneic T cell responses was examined by mixed lymphocyte reaction（MLR）. BALB/c mice were randomly divided into 4 groups. A. Asthma model group, B. Humulus Scandens pollen allergen-specific tDCs treatment group, C. Dexamethasone （DXM） treatment group, D. The normal control group. Humulus pollen allergen was used to establish allergic asthma models. The pathological changes in lung tissues were assayed by HE staining. The numbers of inflammatory cells and percentage of eosinophils（EOS） in bronchoalveolar lavage fluid （BALF） were counted. The percentage of CD4＋ CD25- Foxp3＋ Treg cells was assessed by flow cytometry analysis The level of serum IgE, IFN-γ and IL-4 in culture supernatant of splenocytes and BALF were measured by ELISA method. Results Humulus scandens pollen allergen tolerated dendritic cells can effectively inhibit the infiltration of EOS and accumulation of airway goblet cells in lung tissues, reduce the total numbers of inflammatory ceils,percentage of EOS in BALF, the levels of serum IgE and IL-4 in culture supernatant of splenocytes, and improve the percentage of CD4＋ CD25＋ Foxp3＋ Treg cells. Conclusions Humulus pollen allergen- specific tDCs can protect mice from the allergic airway inflammation.

关 键 词：葎草花粉 哮喘 免疫耐受 树突状细胞 

分 类 号：R562.25[医药卫生—呼吸系统]