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目的分析哮喘、鼻炎、哮喘并发鼻炎患儿以及健康儿童外周血辅助性T细胞(T helper cell,Th)17及其相关细胞因子介导的免疫应答差异。方法流式细胞术检测哮喘组患儿(AS组,n=41)、鼻炎组患儿(AR组,n=40)、哮喘并发鼻炎组患儿(ASR组,n=52)和健康对照组儿童(HC组,n=45)外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中Th1细胞(CD4~+IFN-γ~+T)、Th17细胞(CD4+IL-17A+T)比例差异;酶联免疫吸附法测定血清及螨蛋白抗原刺激后PBMC培养上清中IL-17A、IL-17E和IL-17F水平。检测Th17细胞亚群及其细胞因子与患儿呼出气一氧化氮(fractional exhaled nitric oxide,FeNO)水平、外周血嗜酸粒细胞(eosinophils,EOS)计数间相关性关系。结果外周血Th1细胞比例在哮喘并发鼻炎组、鼻炎组和哮喘组分别为13.2%±5.3%、12.2%±6.7%和12.1%±4.1%,均低于健康对照组(17.0%±5.1%),四组间Th1细胞比例差异具有统计学意义(P<0.05)。外周血Th17细胞比例分别为哮喘并发鼻炎组1.8%±0.7%、哮喘组1.2%±0.5%、鼻炎组1.1%±0.6%和健康对照组0.5%±0.4%,四组间差异有统计学意义(P<0.05)。三组患儿外周血IL-17A、IL-17E水平均显著高于健康对照组,差异具有统计学意义(均P<0.05)。尘螨蛋白抗原刺激后,哮喘并发鼻炎组、哮喘组、鼻炎组患儿PBMC中IL-17E水平显著升高。哮喘组、鼻炎组、哮喘并发鼻炎组患儿外周血IL-17E水平与Fe NO水平、外周血EOS呈正相关关系(均P<0.05),仅发现哮喘并发鼻炎组患儿外周血Th17细胞比例与FeNO水平、外周血EOS增多存在正相关关系(P<0.05)。结论 Th17细胞及其产生的前炎症因子IL-17E参与了哮喘、鼻炎以及哮喘并发鼻炎等气道过敏性疾病的免疫病理过程。
Objective To analyze the difference of T helper cell (Th) 17 in peripheral blood of children with asthma, rhinitis, asthma complicated with rhinitis and healthy children and the related cytokine-mediated immune responses. Methods The flow cytometry was used to detect the expression of IL-6 in asthmatic children (AS group, n = 41), rhinitis group (n = 40), asthmatic rhinitis group (n = 52) and healthy asthmatic children The differences in the proportion of Th1 cells (CD4 ~ + IFN-γ ~ + T) and Th17 cells (CD4 + IL-17A + T) in peripheral blood mononuclear cells (PBMC) of children (HC group, n = 45) The levels of IL-17A, IL-17E and IL-17F in the culture supernatant of PBMC stimulated by serum and mite protein antigen were measured by enzyme-linked immunosorbent assay. To detect the correlation between Th17 cell subsets and their cytokines and the level of exhaled nitric oxide (FeNO) and eosinophils count (EOS) in children. Results The proportion of Th1 cells in peripheral blood was 13.2% ± 5.3%, 12.2% ± 6.7% and 12.1% ± 4.1% in asthma group, rhinitis group and asthma group, respectively, which were lower than those in healthy control group (17.0% ± 5.1% There was a significant difference in the proportion of Th1 cells between the four groups (P <0.05). The proportions of Th17 cells in peripheral blood were 1.8% ± 0.7% in asthmatic group, 1.2% ± 0.5% in asthmatic group, 1.1% ± 0.6% in rhinitis group and 0.5% ± 0.4% in healthy control group respectively. The differences among the four groups were statistically significant (P <0.05). The levels of IL-17A and IL-17E in the peripheral blood of the three groups were significantly higher than those in the healthy control group (all P <0.05). After dust mite protein antigen stimulation, IL-17E levels in PBMC of asthmatic children with rhinitis, asthma and rhinitis were significantly increased. The levels of IL-17E in peripheral blood of asthmatic group, rhinitis group and asthmatic rhinitis group were positively correlated with FeNO level and peripheral blood EOS (all P <0.05). Only the percentage of Th17 cells in peripheral blood of children with asthma complicated with rhinitis There was a positive correlation between FeNO level and peripheral blood EOS (P <0.05). Conclusion Th17 cells and its proinflammatory cytokine IL-17E are involved in the immunopathological process of airway allergic diseases such asthma, rhinitis and asthma complicated with rhinitis.