黄花蒿花粉变应原致敏小鼠鼻炎模型建立及免疫学研究  

Establishment of allergic rhinitis mouse model sensitized by Artemisia annua and its immunological study

作　　者：於洋 张影[2] 鲁旭[2] 周晨昱 杨蕾[2] 曾明[2] 张慧云 徐苗[2] 何韶衡 Yu Yang;Zhang Ying;Lu Xu;Zhou Chenyu;Yang Lei;Zeng Ming;Zhang Huiyun;Xu Miao;He Shaoheng(Research Centre of Allergy and Clinical Immunology,First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,China;Key Laboratory of Quality Research and Evaluation of Biological Products,China Institute for Food and Drug Control,Beijing 100050,China)

机构地区：[1]锦州医科大学附属第一医院变态反应与临床免疫研究中心,121001 [2]中国食品药品检定研究院生物制品质量研究与评价重点实验室,北京100050

出　　处：《中华微生物学和免疫学杂志》2021年第7期516-523,共8页Chinese Journal of Microbiology and Immunology

基　　金：辽宁省高等学校攀登学者(辽教函(2018)38号)。

摘　　要：目的优化黄花蒿花粉变应原致敏BALB/c小鼠鼻炎模型,探索可用于过敏反应评价的体液免疫和细胞免疫指标。方法以BALB/c小鼠为实验动物,以黄花蒿花粉变应原提取物为致敏蛋白,通过不同含量的主要变应原(Art a1)和不同致敏次数,设置不同免疫程序,皮下免疫小鼠,末次免疫后1周和5周分别用含50μg/ml和500μg/ml Art a1的黄花蒿花粉变应原提取物滴鼻激发,每天1次,每次持续1周。观察小鼠的过敏反应,检测鼻部组织病理变化,测定各组小鼠血清中抗原特异性IgE、IgG1、IgG2a等抗体水平及动态变化,检测小鼠脾脏中抗原特异性IL-4、IL-5、IL-2、IFN-γ等淋巴细胞数量变化。结果致敏小鼠经抗原激发后出现明显的抓挠和喷嚏反应;鼻部组织出现明显的变应性炎症;血清中黄花蒿花粉特异性IgE抗体水平的增高与激发抗原呈明显的相关性;致敏小鼠脾脏中抗原特异性IL-4淋巴细胞数量明显增多,而IFN-γ特异性淋巴细胞数量未见显著变化。结论成功建立黄花蒿花粉变应原过敏小鼠模型,通过ELISPOT技术检测到黄花蒿过敏小鼠抗原特异性IL-4淋巴细胞数量的升高,为后续脱敏治疗用制剂的疗效评价提供实验资料。Objective To optimize the BALB/c mouse rhinitis model sensitized by Artemisia annua pollen allergen,and explore the humoral and cellular immune indicators that can be used for the evaluation of allergic reactions.Methods Using BALB/c mice as experimental animals,using Artemisia annua pollen allergen extract as sensitizing protein,through different content of the main allergen Art a1 and different sensitization times,different immunization programs were set to immunize mice subcutaneously,One week and five weeks after the last immunization,Artemisia annua pollen allergen extract containing 50μg/ml and 500μg/ml Art a1 was used for nasal stimulation,once a day,for 1 week each time.Observe the allergic reaction of mice,detect the pathological changes of nasal tissues,determine the levels and dynamic changes of antigen-specific IgE,IgG1,IgG2a and other antibodies in the serum of each group of mice.and detect the changes in the number of antigen-specific IL-4,IL-5,IL-2,IFN-γand other lymphocytes in the spleen of mice.Results Sensitized mice showed obvious scratching and sneezing reactions after being stimulated by antigen;obvious allergic inflammation appeared in nasal tissue;The increase in serum level of Artemisia annua pollen-specific IgE antibody was significantly correlated with the challenge antigen;The antigen-specific IL-4 lymphocytes in the spleen of the sensitized mice were significantly increased,but the IFN-γ-specific lymphocytes did not change significantly.Conclusions The successful establishment of a mouse model of Artemisia annua pollen allergen allergy is the first domestic use of ELISPOT technology to detect an increase in the number of antigen-specific IL-4 lymphocytes in Artemisia annua allergy mice,laying a foundation for the subsequent evaluation of the efficacy of preparations for desensitization treatment basis.

关 键 词：黄花蒿 过敏性鼻炎 动物模型 过敏 

分 类 号：R765.21[医药卫生—耳鼻咽喉科] R-332[医药卫生—临床医学]