摘要
目的探讨外周血CD_(4)^(+)T淋巴细胞及其亚型(Th1/Th2)细胞在初治肺结核患者中的改变及临床意义,并了解其在结核病治疗过程中的变化。方法利用流式细胞术对105例初治肺结核患者、25名正常人外周血CD_(4)^(+)T淋巴细胞进行直接计数,同时用25ng/ml佛波酯(PMA)、250ng/ml离子霉素(ionomycin)对外周血细胞进行刺激,用2μmol/ml莫能霉素(monensin)作蛋白转运抑制剂,5%CO_(2)培养40~45h,而后分别配对加入CD_(3)^(-PC5)+CD_(8)^-(FITC)+INFγPE/CD_(3)^(-PC5)+CD_(8)^-(FITC)+IgG1 PE、CD_(3)^(-PC5)+CD_(8)^-(FITC)+IL-4^(-PE)/CD_(3)^(-PC5)+CD_(8)^-(FITC)+IgG1 PE(PC5:藻红蛋白花青苷5;FITC:异硫氰酸胍荧光素;PE:藻红蛋白)单克隆抗体进行胞膜和胞内标记,进行流式细胞术检测Th1/Th2的百分含量。按病变程度分为重度32例,中度44例,轻度29例。并对其中67例肺结核患者在化疗强化期末、第6个月化疗结束时进行同样指标的追踪检测,以了解在抗结核治疗过程中肺结核患者外周血CD_(4)^(+)T细胞、Th1/Th2细胞的变化状况。结果(1)初治肺结核患者外周血CD_(4)^(+)T细胞、Th1细胞水平显著低于健康对照组,其检测值分别为(663±160)个/μl(9.56±3.60)%和(735±156)个/μl、(18.70±5.03)%(P值均<0.05);(2)重度肺结核患者CD+4 T细胞和Th1细胞的检测值为(579±120)个/μl和(5.43±2.33)%,显著低于轻、中度肺结核患者的(726±166)个/μl和(12.2±1.81)%、(684±192)个/μl和(10.9±2.30)%(P值均<0.01);Th2细胞的水平则显著高于轻、中度肺结核患者,其检测值分别为(5.63±1.26)%、(2.93±0.87)%和(3.22±1.01)%(P值均<0.01);(3)67例随访患者中,61例抗结核治疗后明显好转,外周血CD_(4)^(+)T细胞、Th1细胞的水平增加,Th2细胞的水平减少;(4)痰菌阳性患者与痰菌阴性患者比较,前者Th2细胞的水平显著高于后者,检测值分别为(5.20±0.97)%、(2.77±1.96)%(P<0.05).结论对肺结核患者外周血CD+4 T细胞、Th1/Th2细胞的检测有助于病情判断和疗效观察。
Objective To study the change of peripheral blood CD_(4)^(+)helper T cells and T helper subtypeⅠandⅡ(Th1/Th2)in patients with pulmonary tuberculosis and to explore their changes during anti-tuberculosis chemotherapy.Methods CD_(4)^(+)T cells from 105 patients with pulmonary tuberculosis and 25 normal controls were counted by Flow Cytometry.Peripheral blood cells were stimulated in vitro by phorbol myristate acetate(PMA)(25 ng/ml)and ionomycin(250 ng/ml).Cytokines were confined to the cells by using the protein transporting inhibitor containing monensin(2μmol/ml).These cells were incubated in 5%CO_(2)for 4 h-4.5 h.The membrane and plasma of CD_(4)^(+)helper T cells were marked by CD_(3)^(-PC5)+CD_(8)^-(FITC)+INF-γ-PE/CD_(3)^(-PC5)+CD_(8)^-(FITC)+IgG 1-PE、CD_(3)^(-PC5)+CD_(8)^-(FITC)+IL-4^(-PE)/CD_(3)^(-PC5)+CD_(8)^-(FITC)+IgG_(1)^(-PE)monoclonal antibodies respectively.Th1 and Th2 cells were counted and the ratio of Th1/Th2 cells was calculated.The levels of Th1 and Th2 cells in the 67 patients with pulmonary tuberculosis were detected at the end of intensive chemotherapy and at the sixth month of chemotherapy.Results(1)The levels of CD_(4)^(+)T and Th1 cells in patients with tuberculosis were significantly lower than those of controls.Their values were(663±160)/μl vs(735±156)/μl and(9.56±3.60)%vs(18.7±5.03)%respectively(P<0.05).(2)The levels of CD+4T and Th1 cells in patients with severe pulmonary tuberculosis were lower than those in patients with moderate or mild pulmonary tuberculosis.Their values were(579±120)/μl vs(726±166)/μl,(684±192)/μl and(5.43±2.33)%vs(12.2±1.81)%and(10.9±2.30)%respectively(P<0.05).The level of Th2 cells was in contrast to Th1 cells and their values were(5.63±1.26)%vs(2.93±0.87)%and(3.22±1.01)%(P<0.01).(3)The level of Th1 cells increased while that of Th2 decreased in the 61 patients who gradually recovered.(4)The level of Th2 cells in patients with smear positive tuberculosis was strikingly higher than that in patients with smear negative tuberculosis and their values were(5.20±0.97)%vs(2.77±1.96)%(P<0.05).Conclusion The detection of CD_(4)^(+)helper T cells and Th1/Th2 cells in the peripheral blood cells from patients with pulmonary tuberculosis is useful in evaluating the state of disease and the effect of chemotherapy.
作者
谭耀驹
张言斌
冯蝶仪
谢贝
林兆源
陈洪光
李一耕
TAN Yao-ju;ZHANG Yan-bin;FENG Die-yi;XIE Bei;LIN Zhao-yuan CHEN Hong-guang;LI Yi-geng(Pulmonary Disease Institute,Guangzhou Chest Hospital,Guangzho 510095,China)
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2004年第6期385-389,共5页
Chinese Journal of Tuberculosis and Respiratory Diseases
基金
广东省科技厅资助项目[粤科计2000-261]
