摘要
目的检测特异性免疫治疗前后,桦树花粉诱导的哮喘小鼠脾Th17细胞数量和其转录因子Rorc表达变化。方法小鼠采用颈后皮下注射(桦树花粉和佐剂)致敏和雾化激发的方法造模。通过检测哮喘相关指标评价动物模型和特异性免疫治疗疗效,检测指标包括气道高反应,肺组织病理和血清免疫球蛋白水平(sIgE、sIgG1、sIgG2a)等。流式细胞术检测Th17细胞;RT-qPCR检测Rorc mRNA表达;高通量多因子检测平台Luminex 200TM检测血清细胞因子IL-4和IL-17A水平。结果与对照组比较,桦树花粉致敏和激发的哮喘小鼠具有气道高反应;血清桦树花粉特异性sIgE,sIgG1,sIgG2a水平升高;支气管周围有炎症细胞浸润,支气管内有大量黏液分泌;血清细胞因子IL-4和IL-17A水平明显升高。经特异性免疫治疗后,以上各项指标均明显下降。哮喘组小鼠脾Th17细胞数量(0.37±0.16)较对照组小鼠脾Th17细胞数量(0.18±0.86)明显增高(P<0.05);而特异性免疫治疗组,Th17细胞数量(0.16±0.16)较哮喘组显著降低(P<0.01)。哮喘组小鼠脾Th17细胞转录因子Rorc的表达量(3.99±1.39)较对照组(1.79±1.19)显著升高(P<0.01);经特异性免疫治疗后,Th17细胞转录因子Rorc的表达量(0.65±0.23)较哮喘组明显降低(P<0.01),差异有统计学意义。结论桦树花粉特异性免疫治疗能部分抑制Th17细胞的数量和功能,有助于进一步了解特异性免疫治疗的机制。
Objective To investigate the changes of Th17 cells and its transcription factor Rorc in spleen of asthmatic mice before and after subcutaneous immunotherapy (SCIT). Methods The mice were sensitized by subcutaneous injection (birch pollen with alum adjuvant) and challenged by birch pollen aerosol. Then, the asthma condition and therapeutic effect of SCIT was evaluated by using a series of evaluating indicators, such as airway responsiveness, histological analysis, and immunoglobulins (sIgE, sIgG1, sIgG2a). Th17 cells were detect by Flow Cytometry and Rorc mRNA levels was detect by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The serum IL-4 and IL-17A levels were determined by the MILLIPLEX MAP Mouse Cytokine/ Chemokine Magnetic Panel. Results Airway hyper-responsiveness (AHR) developed in birch pollen sensitized and challenged mice, compared to control group. Serum birch pollen specific IgE, IgG1, and IgG2a levels increased in asthmatic mice. Peribranchial infiltration of inflammatory cells was shown among asthmatic mice, with enhanced mucus production. Serum IL-4 and IL-17A levels were also higher in asthmatic mice. All the parameters decreased significantly after SCIT. The number of Th17 cells in the spleen of asthmatic mice (0.37±0.16) was higher than control (0.18±0.86) and SCIT group (0.16±0.16). The level of transcription factor Rorc mRNA also increased significantly in the spleen of asthmatic mice(3.99±1.39) than control(1.79±1.19) and SCIT group(0.65±0.23). Conclusion Birch pollen SCIT can partially inhibit Th17 related immune response in mice.
作者
谢枝隽
尹佳
周俊雄
XIE Zhi-juan;YIN Jia;ZHOU Jun-xiong(Department of Allergy, Beijing Key Laboratory of Precision Medicine for Diagnosis and Treatment on Allergic Diseases,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730,China)
出处
《中华临床免疫和变态反应杂志》
2019年第2期130-136,共7页
Chinese Journal of Allergy & Clinical Immunology
