肺结核患者外周血中Th细胞极化偏移及临床意义分析  

Study on T helper cells in the patients of pulmonary tuberculosis and the clinical significance of Th polarization

作　　者：胥萍[1] 施美华[1] 费晓峰[1] 朱传武[1] 吴妹英[1] 吴敏娟[1] 陈永井[2] 张学光[2] 

机构地区：[1]苏州市第五人民医院中心实验室,苏州215007 [2]苏州大学医学生物技术研究所,苏州215007

出　　处：《中国免疫学杂志》2010年第2期178-181,185,共5页Chinese Journal of Immunology

基　　金：苏州市科学技术局资助项目[苏财科字(2005)47号]

摘　　要：目的:分析初诊肺结核患者外周血中CD4+T淋巴细胞及其亚型Th1和Th2细胞的改变,探讨其在肺结核病变过程中的临床意义。方法:取肝素抗凝全血,加RPMI1640培养液等体积混匀,依次加入1∶1 000稀释的PMA、1∶100稀释的Ion-omycin和1∶10稀释的Monensin,混匀后于37℃,5%CO2静置培养4小时或过夜。取100μl培养血细胞依次加入抗人CD3-Per-CP、CD8-APC、mIgG1-FITC、Rat IgG1-PE、IL-4-PE、IFN-γ-FITC抗体,按流式操作流程分别进行细胞膜和细胞浆内标记测定CD4+IL-4+(Th2)、CD4+IFN-γ+(Th1)两种细胞的水平。结果:初诊肺结核患者外周血中Th1水平均显著低于健康对照组(P<0.01),而Th2水平则显著高于健康对照组(P<0.05)。粟粒型肺结核患者外周血中Th1细胞含量显著低于浸润性肺结核患者(P<0.05),而Th2水平显著高于浸润性肺结核和结核性胸膜炎(P<0.05)。CD4+/CD3+T细胞比例在这三个病程中呈下降趋势,且粟粒型肺结核显著低于浸润性肺结核(P<0.05)。糖尿病并肺结核患者Th1、CD4+/CD3+显著低于无糖尿病肺结核患者(P<0.05),Th2含量则显著升高(P<0.05)。15例重度肺结核患者经结核化疗与微卡治疗三个月后Th1、CD4+/CD3+水平较治疗前明显升高(P<0.05),而Th2水平较治疗前显著降低(P<0.01)。痰检或培养阳性与痰检阴性患者相比Th1、CD4+/CD3+水平呈下降趋势但无显著差异(P>0.05),Th2细胞水平显著升高(P<0.05)。结论:浸润性肺结核、结核性胸膜炎、粟粒型肺结核、糖尿病并肺结核患者存在着不同程度的免疫功能抑制,对Th1、Th2细胞水平与CD4+/CD3+比例测定有助于临床病情的判断和疗效观察。Objective：This study was to analyze the changes of CD4＋ T lymphocytes and their subtypes,Thl and Th2 cells, in the peripberal blood of patients with pulmonary tuberculosis disease and to investigate their clinical significance in the pathologic process of pulmonary tuberculosis. Methods： For polarization measurement of T-helper cells, 1：100 diluted Ionomycin and 1：10 diluted Monensin were added in sequence into the equivalent volume mixture of heparin anti-coagalated whole blood and RPMI-1640 culture liquid. After being well mixed, the mixture was incubated at 5 % CO2,37℃ for 4 hours or overnight. To 100 μl of the mixture and in sequence, the antibodies of CD3-PerCP, CD8- APC,mIgG1-FITC,Rat IgG1-PE,IL-4-PE or IFN-γ-FITC were added. The stained CD4＋ IL4＋ （Th2） and CD4＋ IFN-γ （Thl） were analyzed by flow cytometry. Results： Compared with those from healthy controls, the peripheral blood of pulmonary tuberculosis patients contained significantly fewer Thl （ P 〈 0.01） but significantly more Th2 cells （ P 〈 0.05） .Thl cells in the peripheral blood of the patients with mitiary pulmonary tuberculosis were obviously fewer （P 〈 0.05） than in infiltrative pulmonary tuberculosis patients. The amount of Th2 ceils in the peripheral blood of the patients with mihary pulmonary tuberculosis was significantly more （ P 〈 0.05） than in either infiltrative pulmonary tuberculosis or tuberculous pleurisy patients. The ratio of CD4 ＋/CD3 ＋tended to decrease in all these patients, and it was much lower （P 〈 0.05） in the patients of miliary pulmonary tuberculosis than in infiltrative pulmonary tuberculosis patients. Patients suffering from both pulmonary tuberculosis and diabetes had significantly lower levels of Thl cells and CD4＋/CD3 ＋ （ P 〈 0.05） and more Th2 cells, compared with those of pulmonary tuberculosis patients without diabetes, levels of Thl and Th2 ceils restored significantly （P 〈 0.05） in 15 severe pulmonary tuber- culosis patients after rece

关 键 词：肺结核 细胞免疫 TH细胞 细胞极化 

分 类 号：R521[医药卫生—内科学] R392.12[医药卫生—临床医学]