肺结核患者CD3^+ CD56^+ NKT细胞对IL-2刺激的反应性研究

IL-2 stimulated responses of CD3^+ CD56^+ NKT cells in pulmonary tuberculosis patients

目的:观察IL-2刺激后肺结核患者CD3+CD56+NKT细胞活化的规律和增殖情况。方法:结核病患者和健康人外周血单个核细胞用IL-2刺激和培养不同时间,采用多色荧光染色和流式细胞术检测NKT细胞的CD69的表达率以及培养前后NKT细胞数量的变化。结果:肺结核患者外周血NKT细胞的比率在刺激培养前与正常人比较无明显差异;经IL-2刺激0、8、16、40和64 h后,患者组和正常组NKT细胞表面CD69的表达量均显著增加,但患者组NKT细胞的CD69表达水平高于正常组(P<0.05);培养14 d后,患者组NKT细胞的数量扩增(108.69±59.22)倍,正常组NKT细胞的数量扩增(246.26±134.06)倍,明显高于患者组(P<0.05)。结论:肺结核患者外周血NKT细胞在IL-2刺激后表现为高度活化和低增殖的特点。

AIM： To observe the activation and proliferation characteristics of IL-2 stimulated CD3＋CD56＋ NKT cells in pulmonary tuberculosis（PTB） patients.METHODS： Peripheral blood mononuclear cells（PBMCs） from PTB patients and normal subjects were stimulated with IL-2 and cultured for different time points.The CD69 expression on and amount of the CD3＋CD56＋ NKT cells were detected by multi fluorescence staining and flow cytometry at different time of stimulation and culture.RESULTS： There was no significant difference in percentage of NKT cells between PTB patients and normal healthy controls before culture.When IL-2 was used to stimulate for 0 h,8 h,16 h,40 h and 64 h,the expression of CD69 on NKT cells in normal controls and PTB patients increased significantly,but the CD69 expression level of NKT cells in PTB patients was significantly higher than that in normal persons（P0.05）.In PTB patients group,PBMCs were expanded significantly after stimulated by IL-2,the absolute number of NKT cells increased from（3.44±1.20）×104 to（323.23±75.98） ×104（P0.01）,expanded by 108.69±59.22 fold,In normal control group the absolute number of NKT cells increased from（5.57±5.16）×104 to（1475.05±868.98）×104（P0.01）,expanded by 246.26±134.06 fold,the expanding efficiency in PTB group was significantly lower than that in normal control group（P0.05）.CONCLUSION： NKT cells in PTB patients present with high activation but low proliferation after stimulated by IL-2.