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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

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50 μg/ml和500 μg/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。

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abstractsObjective: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 50 μg/ml and 500 μg/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-γ and 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.

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