变应性鼻炎小鼠模型中外周血及脾脏Tfh细胞的流式细胞分析  

Flow cytometry analysis of circulating and splenic follicular helper T （ Tfh） cells in a mouse model of allergic rhinitis

作　　者：刘志奇[1] 杨琨[2] 刘立思 

机构地区：[1]湖北省妇女儿童医院耳鼻咽喉科,武汉430075 [2]武汉大学人民医院耳鼻咽喉头颈外科,430060 [3]华中科技大学同济医学院附属同济医院耳鼻咽喉头颈外科,武汉430030

出　　处：《中华微生物学和免疫学杂志》2017年第11期834-839,共6页Chinese Journal of Microbiology and Immunology

摘　　要：目的 探讨滤泡辅助性T细胞（Tfh）在变应性鼻炎小鼠模型外周血及脾脏中的数量变化和意义.方法 采用卵清蛋白（OVA）及氢氧化铝致敏BALB/c小鼠建立变应性鼻炎模型.Tfh细胞用抗小鼠CD4-FITC和CXCR5-PE染色,采用流式细胞仪计数得到CD4＋CXCR5＋在小鼠外周血及脾脏中的比例.采用酶联免疫吸附试验测定血清总OVA特异性IgE（OVA-sIgE）的水平.检测结果用统计学软件SPSS16.0分析.结果 在实验组中,鼻黏膜下层嗜酸性粒细胞浸润增多;外周血OVA-sIgE浓度增加,对照组为43.47 pg/ml[（43.47±2.58）pg/ml],实验组为50.44 pg/ml[（50.44±1.40）pg/ml]（P=0.029）;对照组外周血Tfh百分比为6.25％[（6.25±1.19）％],实验组为13.94％[（13.94±2.77）％]（P=0.026）;对照组脾脏Tfh百分比为18.04％[（18.04±4.97）％],实验组为7.26％[（7.26±0.96）％]（P=0.019）.Pearson相关分析显示外周血Tfh细胞的数量与OVA-sIgE浓度呈正相关（r=0.954,P=0.003）,与脾脏中Tfh的数量呈显著性负相关（r=-0.801,P=0.028）,外周血Tfh细胞的数量与脾脏中的Tfh细胞显著性负相关（r=-0.787,P=0.032）.结论 Tfh细胞可能参与了小鼠变应性鼻炎的免疫过程.Objective To investigate changes in the percentages of circulating and splenic follicular helper T （ Tfh） cells and the significance of Tfh cells in a mouse model of allergic rhinitis .Meth-ods BALB/c mice were sensitized by injection of OVA and aluminum hydroxide to establish the model of allergic rhinitis .Flow cytometry was performed to measure the percentages of circulating and splenic CD 4＋CXCR5＋after staining Tfh cells with anti-mouse CD4-FITC and anti-mouse CXCR5-PE.Concentrations of OVE-specific IgE （ OVA-sIgE ） in serum samples were measured by enzyme-linked immunosorbent assay （ ELISA ） .Student′s t-test and Pearson ′s correlation analysis were used for statistical analysis .Results Enhanced infiltration of eosinophils in submucosa was observed in the experimental group .After sensitizing the mice with OVA and aluminum hydroxide , the concentration of OVA-sIgE increased from 43 .47 pg/ml [（43.47±2.58） pg/ml] to 50.44 pg/ml [（50.44±1.40） pg/ml] （P=0.029）; the percentage of circu-lating CD4＋CXCR5＋was up-regulated from 6.25% [（6.25±1.19）%] to 13.94% [（13.94±2.77）%] （P=0.026）; the percentage of splenic CD4＋CXCR5＋was down-regulated from 18.04% [（18.04 ± 4.97）%] to 7.26%[（7.26±0.96）%] （P=0.019）.The concentration of OVA-specific IgE was posi-tively correlated with the percentage of circulating Tfh cells （r=0.954, P=0.002）, but negatively correla-ted with the percentage of splenic Tfh cells （r=-0.801, P=0.028）.Moreover, a negative correlation was found between circulating and splenic Tfh cells （r=-0.787, P=0.032）.Conclusion Tfh cells might be involved in the immune responses against allergic rhinitis in mice .

关 键 词：滤泡辅助性T细胞 变应性鼻炎 流式细胞术 免疫球蛋白E 小鼠 

分 类 号：R-332[医药卫生] R765.21