初治肺结核患者外周血Th1/Th2细胞的测定及临床意义

The change and the clinical significance of peripheral blood Th1/Th2 cells in patients with pulmonary tuberculosis

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 10 5例初治肺结核患者、2 5名正常人外周血CD+ 4 T淋巴细胞进行直接计数 ,同时用 2 5ng/ml佛波酯 (PMA)、2 5 0ng/ml离子霉素 (ionomycin)对外周血细胞进行刺激 ,用 2 μmol/ml莫能霉素 (monensin)作蛋白转运抑制剂 ,5 %CO2培养 4 0～ 4 5h ,而后分别配对加入CD3 PC5+CD8 FITC+INF γ PE/CD3 PC5+CD8 FITC+IgG1 PE、CD3 PC5+CD8 FITC+IL 4 PE/CD3 PC5+CD8 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 4 4例 ,轻度 2 9例。并对其中 6 7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 (6 6 3± 16 0 )个 / μl、(9 5 6±3 6 0 ) %和 (735± 15 6 )个 / μl、(18 70± 5 0 3) % (P值均 <0 0

Objective To study the change of peripheral blood CD+ 4 helper T cells and T helper subtype Ⅰand Ⅱ(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 (2 μmol/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.