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目的观察经屋尘螨特异性免疫治疗(SIT)的哮喘患儿对尘螨过敏原特异性免疫反应的变化。方法选取54例尘螨过敏性哮喘患儿,其中男性38例,女性16例;年龄6~11岁,平均年龄9岁1个月。27例经标准化屋尘螨过敏原皮下SIT疗程1.5~2.0年患儿作为SIT组,另27例未经SIT患儿作为对照组。应用最大呼气流量-容积曲线测定方法测定肺通气功能,参数为第1秒用力呼气容积(FEV1)占预计值百分比(FEV1%);应用荧光酶联免疫法测定2组患儿血清屋尘螨特异性IgE和特异性IgG4浓度;分离患儿外周血单个核细胞(PBMC),经质量浓度20μg/mL屋尘螨蛋白浸液刺激并培养48 h后,检测CD4+CD25+Foxp3+调节性T淋巴细胞(Treg细胞)的相对数量和可分泌白细胞介素10(IL-10)的Treg细胞百分比;同时留取细胞培养上清液应用酶联免疫吸附分析检测细胞因子IL-4、干扰素-γ(IFN-γ)和转化生长因子β1(TGF-β1)水平。结果 SIT组患儿肺功能FEV1%与对照组相比,差异无统计学意义(P>0.05);SIT组IgG4水平显著高于对照组,差异有极显著统计学意义(P<0.000 1)。SIT组IgE水平与对照组相比,差异无统计学意义(P>0.05);两组刺激后Treg细胞百分数分别与各自该组内未经刺激时基础水平的差异无统计学意义。两组间PBMC无论是否经屋尘螨蛋白浸液刺激时Treg细胞百分数的差异无统计学意义(P>0.05);SIT组刺激后Treg细胞百分数高于自身未经刺激基础水平1.56倍。对照组经刺激后与未接受刺激比较,Treg细胞百分数无显著变化;SIT组PBMC培养上清液中IL-4和TGF-β1浓度显著低于对照组,IFN-γ浓度显著高于对照组,差异有统计学意义(P<0.05)。结论屋尘螨SIT的哮喘患儿对尘螨过敏原特异性免疫反应表现为Treg细胞比率增加,Th1/Th2免疫反应平衡改变,屋尘螨特异性IgG4浓度增高,提示发生对尘螨过敏原特异性免疫耐受的特征。
Objective To observe the changes of specific immune responses to dust mite allergens in asthmatic children with house-dust mite-specific immunotherapy (SIT). Methods 54 cases of dust mite allergic asthma children, including 38 males and 16 females; aged 6 to 11 years, with an average age of 9 years and 1 month. 27 cases of standardized house dust mite allergens subcutaneous SIT treatment of 1.5 to 2.0 years of children as SIT group and the other 27 cases without SIT children as a control group. The maximum expiratory volume-volumetric curve method was used to measure the pulmonary ventilation function, the parameters were forced expiratory volume in 1 second (FEV1) percentage of predicted value (FEV1%); serum enzyme-linked immunosorbent assay Mite-specific IgE and specific IgG4 concentration. Peripheral blood mononuclear cells (PBMCs) from children were isolated and stimulated with protein extract of house dust mite (20μg / mL) for 48 h. CD4 + CD25 + Foxp3 + regulatory T Lymphocytes (Treg cells) and the percentage of Treg cells that can secrete interleukin-10 (IL-10). Meanwhile, the cell culture supernatants were collected for detection of cytokines IL-4, IFN- γ (IFN-γ) and transforming growth factor β1 (TGF-β1) levels. Results There was no significant difference in lung function FEV1% between SIT group and control group (P> 0.05). The level of IgG4 in SIT group was significantly higher than that in control group (P <0.0001). The level of IgE in SIT group was not significantly different from that in control group (P> 0.05). There was no significant difference between the two groups in the percentage of Treg cells and the basal level in each group without stimulation. There was no significant difference in the percentage of Treg cells between the two groups (P> 0.05). The percentage of Treg cells in SIT group was 1.56 times higher than that of SIT group. The percentage of Treg cells did not change significantly in the control group compared with the untreated group. The concentrations of IL-4 and TGF-β1 in the culture supernatant of PBMC in SIT group were significantly lower than those in the control group, and the concentration of IFN-γ was significantly higher than that in the control group The difference was statistically significant (P <0.05). Conclusions The specific immune response to dust mite allergens in SIT-induced asthmatic children is characterized by an increase in the proportion of Treg cells, a shift in the Th1 / Th2 immune response, and an increase in the specific IgG4 concentration in house dust mites, suggesting a specific response to dust mite allergens Sexual immune tolerance characteristics.