卡介苗与结核分枝杆菌早期分泌靶抗原刺激人γδT细胞分泌细胞因子能力的比较  

Comparison of abilities of BCG and early secreted antigenic target-6of Mycobacterium tuberculosis in stimulating secretion of cytokines by human γδT cells

作　　者：代光明[1] 汪洪[1] 郑权[1] 杜发旺[2] 杜先智[2] 

机构地区：[1]四川省遂宁市第一人民医院呼吸内科,四川遂宁629000 [2]重庆医科大学附属第二医院呼吸内科,重庆400010

出　　处：《中国生物制品学杂志》2013年第11期1567-1570,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金(30872261)

摘　　要：目的比较卡介苗(bacillus Calmette-Guerin vaccine,BCG)与结核分枝杆菌早期分泌靶抗原(early secreted antigenic target-6,ESAT-6)刺激人外周血γδT细胞分泌细胞因子的能力。方法采用Ficoll密度梯度离心法分离健康人外周血单核细胞(peripheral blood mononuclear cell,PBMC),加入Anti-human gamma-delta TCR-FITC抗体标记γδT细胞,上流式细胞仪进行分离纯化;分别用BCG和ESAT-6刺激γδT细胞,同时,以不加任何刺激因子的γδT细胞作为空白对照,分别于刺激培养后第1、3、6、9和12天取上清,采用ELISA试剂盒检测IL-17、TNF-α及IFNγ的分泌水平;上流式细胞仪检测γδT细胞的增殖水平。结果γδT细胞占PBMC的4.83%,其纯度为88.90%;与空白对照组比较,BCG和ESAT-6刺激的γδT细胞分泌的IL-17、TNF-α及IFNγ水平均明显升高,且ESAT-6组明显高于BCG组(P均<0.05);与空白对照组相比,BCG组和ESAT-6组γδT细胞数量明显增加,且ESAT-6组较BCG组增加明显(P均<0.05)。结论 BCG和ESAT-6均可刺激γδT细胞增殖,并大量分泌IL-17、TNF-α及IFNγ,且ESTA-6的刺激作用强于BCG。Objective To compare the abilities of BCG and early secreted antigenic target-6 （ESAT-6） of Mycobacterium tuberculosis in stimulating the secretion of cytokines by human peripheral blood γδT cells. Methods Peripheral blood mononuclear cells （PBMCs） of healthY humans were isolated by Ficoll density gradient centrifugation, in which γδT cells were labeled with anti-human gamma-delta TCR-FITC, purified by flow cytometry, and stimulated with BCG and ESAT-6, using those not stimulated as blank control. Supernatants were collected on days 1, 3, 6, 9 and 12 after stimulation and determined for secretion levels of IL-17, TNF-α and IFNγ by ELISA. The proliferation level of γδT cells was determined by flow cytometry. Results The γδT cells accounted for 4. 83% of PBMCs, of which the purity was 88. 90%. The secre-tion levels of IL-17, TNF-α and IFNγ by γδT cells stimulated with BCG and ESAT-6 increased significantly as compared with those in blank control group （P 〈 0. 05）. However, the secretion level in ESAT-6 group was significantly higher than that in BCG group （P 〈 0. 05）. The count of γδT cells was significantly higher in BCG and ESAT-6 groups than in blank control group, and in ESAT-6 group than in BCG group （both P 〈 0. 05）. Conclusion Both BCG and ESAT-6 stimulated the proliferation of γδT cells and secretions of IL-17, TNF-αand IFNγ. However, the ability of ESAT-6 in stimulating the secretion of cytokines was stronger than that of BCG.

关 键 词：卡介苗 结核分枝杆菌 靶抗原 ΓΔT细胞 细胞因子 

分 类 号：R378.911[医药卫生—病原生物学] R392.11[医药卫生—基础医学]