粉尘螨变应原对支气管哮喘患儿外周血淋巴细胞增殖功能的影响

目的 通过观察粉尘螨(DF)浸出液在体外刺激对粉尘螨过敏的支气管哮喘患儿外周血琳巴细胞增殖情况,确定所需DF浸出液的最佳刺激浓度和刺激时间.方法 支气管哮喘患儿40例分为粉尘螨过敏组20例,非粉尘螨过敏组20例,应用细胞计数试剂盒8(CCK-8)法检测两组患者对DF过敏的中度支气管哮喘患儿外周血淋巴细胞经DF变应原刺激后的增殖情况.计算不同浓度DF浸出液刺激淋巴细胞增殖的刺激指数(SI).结果 DF引起了支气管哮喘患儿淋巴细胞增殖的SI升高.刺激物浓度为20 mg/L、40 mg/L时,DF过敏组在培养24小时、48小时和72小时的过程中,淋巴细胞增殖的SI均呈先升高后降低的趋势,而在刺激物浓度为80 mg/L时,粉尘螨过敏组淋巴细胞增殖的SI则是逐渐降低的；在非粉尘螨过敏组,这种升高和降低的趋势不明显,两组在组间,不同时点以及组间和不同时点的交互作用中差异均有统计学意义(P＜0.05或＜0.01),刺激物浓度为20 mg/L时,粉尘螨过敏组在24、48和72小时的淋巴细胞刺激指数是1.14±0.17、1.43±0.37和0.95±0.34,非粉尘螨过敏组为1.08±0.29,0.99±0.26和0.95±0.24;刺激物浓度为40 mg/L时,粉尘螨过敏组24、48和72小时的淋巴细胞刺激指数是1.34±0.25,1.68±0.32和0.89±0.37;非粉尘螨组为0.96±0.34,0.95±0.34和0.96±0.33,刺激物浓度为80 mg/L时,粉尘螨过敏组24、48和72小时的淋巴细胞刺激指数是1.17±0.13、0.98±0.26和0.79±0.34.非粉尘螨组为0.98±0.29、0.95±0.31和0.91±0.26.粉尘螨浸出液刺激浓度为40 mg/L,培养48小时时淋巴细胞增殖的SI最高(1.68±0.32).结论 刺激淋巴细胞增殖的最佳粉尘螨浸出液刺激浓度和刺激时间为:40 mg/L,刺激48小时.

Study on proliferation of peripheral blood lymphocyte caused by house dust mite in asthmatic children

Objective To investigate the best time and dose of Dermatophagoides farinae （DF） extract by detecting the proliferation of peripheral blood lymphocyte caused by specific allergen in allergic bronchial asthma. Methods Allergic asthmatic children 40 cases were divided into DF allergy group（ n = 20）, none--DF allergy group （n=20）. Stimulation index （SI） of lymphocyte was measured by cell counting Kh-8 （CCK-8） assay in the patients of both groups . Results The SI of lymphocyte was increased by DF in allergic asthmatic children. With DF concentration 20 rag/L, 40 mg/L at the culture for 24,48 and 72 hours, the SI of lymphocyte in DF allergy group was showing a tendency of first rise then fall. While DF concentration at 80 mg/L, the IS in DF allergy group grandually decreased;in none-DF allergy group, the rise and fall trend was not obvious. The two groups showed significant difference in inter- group,time points, mutual action between groups and time points（ P 〈0.05 or 〈0.01）. With the concentration 20 mg/L at the culture for 24,48 and 72 hours,the SI of lymphocyte in DF allergy group was 1.14±0.17 , 1. 43±0.37 and 0.95± 0.34. The SI of lymphocyte in none-DF allergy group was 1.08± 0.29,0.99 ± 0.26 and 0.95± 0.24. Accordingly, the IS was still increased with the treatment of DF （40 rag/L） as compared with the control （1.34!0.25 vs0.96±0.34 for 24 h; 1.68±0.32 vs 0.95±0.34 for 48 h; 0.89±0.37 vs 0.96±0.33 for 72 h）.But,the SI of lymphocyte compared with the control was decreased when the DF concentration at 80 mg/L （1. 17±0.13 vs 0. 98± 0.29 for 24 h; 0. 98±0. 26 vs 0. 95±0.31 for 48 h; 0. 79±0.34 vs 0. 91±0.26 for 72 h）. These results showed that the SI of lymphocyte was the highest with the treatment of DF at 40 mg/L for 48 h. Conclusion The best dose and time of DF extract treatment was 40 mg/L for 48 hour.