肺结核患者疗程中外周血辅助性和调节性T细胞的动态研究

The dynamic change of Th17 cells and regulatory T cells in PBMC during the anti-tuberculosis treatment of the patients with pulmonary tuberculosis

目的探讨辅助性T细胞17(Th17)和CD4+CD25+CD127low调节性T细胞与结核病的发病及抗结核治疗转归的关系。方法纳入对象包括32例活动性肺结核患者、25例Mtb潜伏感染者、45例健康对照者。流式细胞术检测外周血Th17细胞分泌细胞因子IL-17和CD4+CD25+CD127low调节性T细胞表达强度。结果以x±s表示,所有数据均使用Prism 4.0统计软件进行分析,两组间比较采用非配对t检验,多组间的比较采用ANOVA方差分析,以P<0.05为差异有统计学意义。结果肺结核患者组治疗前IL-17表达为(3.25±1.68)%,明显低于健康对照组[(4.62±1.46)%](F=6.633,P<0.0001)。比较活动性肺结核患者组IL-17表达在治疗前、治疗3个月[(4.17±2.27)%]、治疗6个月[(5.58±1.66)%]时的检测结果,发现治疗3个月时高于治疗前,但差异无统计学意义(F=12.244,P=0.057);而治疗6个月时明显高于治疗前(F=12.244,P<0.0001)和治疗3个月时(F=12.244,P=0.004)。健康对照组CD4+CD25+CD127low调节性T细胞表达为(4.97±1.60)%,与Mtb潜伏感染组[(5.00±1.08)%]比较,差异无统计学意义(F=11.986,P=0.937);活动性肺结核患者组治疗前表达为(6.59±1.73)%,显著高于健康对照组及Mtb潜伏感染组(F=11.986,P<0.0001)。比较活动性肺结核患者组治疗前、治疗3个月[(8.28±2.04)%]、治疗6个月[(7.46±1.87)%]时的CD4+CD25+CD127low调节性T细胞表达,发现治疗3个月时的表达明显高于治疗前(F=6.458,P=0.001);治疗6个月时的表达低于治疗3个月时(F=6.458,P=0.085),但差异无统计学意义;治疗6个月与治疗前的表达相比较,差异无统计学意义(F=6.458,P=0.068);治疗6个月CD4+CD25+CD127low调节性T细胞表达明显高于健康对照组(t=6.255,P<0.0001)。结论结核病患者外周血Th17细胞明显减少,经有效抗结核治疗后,Th17细胞逐渐增加,表明Th17细胞在抗结核免疫中起保护作用。结核病患者外周血CD4+CD25+CD127low调节性T细胞明显增多,经抗结核治疗后CD4+CD25+CD127low调节性T细胞逐渐减少,进一步说明CD4+CD25+CD127low调节性T细胞在抗结核免疫中起抑制作用。

Objective To study the relationship between the numbers of Th17cells,CD4 ＋ CD25 ＋ CD127 low Tregs and the tuberculosis（TB） and outcome of anti-TB.Methods Intracellular staining and flow cytometry analysis were used to evaluate IL-17cytokine secreted by Th17cells and the responses of CD4 ＋ CD25 ＋ CD127 low Treg cells in peripheral blood samples collected from 25individuals with LTBI,45healthy donors（HD）,32patients with active pulmonary TB.The results were showed as x±s,all data were analyzed by software Prism 4.0.Two groups were compared by T-test,the comparison between groups used ANOVA.There is statistical significance if P 0.05.Results The percentage of IL-17in TB group [（3.25±1.68） % ] was significantly lower than that in HD group [（4.62±1.46） % ]（F=6.633,P〈0.0001） before treatment.The dynamic variation of IL-17percentage was monitored at different time points ： before treatment [（3.25±1.68） % ],at 3 months [（4.17±2.27） % ],and at 6months after treatment [（5.58±1.66） % ].The percentage of Th17cells from TB group at 3months after treatment was higher than that before treatment,but there was no significant difference（F=12.244,P=0.057）.The percentage of Th17cells at 6 months after treatment was significantly higher than those before treatment（F= 12.244,P〈0.0001） and at 3 months after treatment（F=12.244,P=0.004）.The percentage of CD4 ＋ CD25 ＋CD127 low treg cells in LTBI group [（5.00±1.08） % ] was higher than that in HD group [（4.97±1.60） % ],but there was no significant difference（F=11.986,P=0.937）.The percentage of CD4 ＋ CD25 ＋ CD127 low treg cells in TB group [（6.59±1.73） % ] was significantly higher than that in HD group and LTBI group（F=11.986,P 0.0001）.The Treg cells at 3 months after treatment were significantly higher than that before treatment（F= 6.458,P=0.001）.But the Treg cells at 6months after treatment were lower than that at 3months after treatment（F=6.458,P=0.085）.There was no significant difference between before treatment and at 6months after treatment（F=6.458,P=0.068）.The Treg cells at 6months after treatment were significantly higher than HD group（t=6.255,P〈0.0001）.Conclusion The Th17cells in TB group were low,gradually increased after effective anti-TB treatment,which suggested that Th17cells play aprotective role in the anti-TB immunity.The CD4 ＋ CD25 ＋ CD127 low treg cells in TB group were higher,and gradually reduced after effective anti-TB treatment,which suggested that CD4 ＋ CD25 ＋ CD127 low treg cells play a supressive role in the anti-TB immunity.