摘要
目的 寻找一种准确、敏感的检测 T淋巴细胞内细胞因子水平的方法 ,为研究 T细胞极化状态在疾病发生发展过程中的作用提供可靠的手段。 方法 用 PMA、 ionomycin作刺激剂 ,刺激全血中淋巴细胞表达细胞因子 ,以 mon-ensin阻断细胞因子分泌至胞外 ,应用两种免疫荧光抗体同时标记淋巴细胞的膜表面特异的分子和表达并被阻滞在胞内的细胞因子 ,然后用流式细胞仪进行检测和分析。 结果 本方法检测 1 8例正常人淋巴细胞的分泌细胞因子水平 ,其中CD4 + T淋巴细胞中 Th1占 1 9.5 1 %± 4 .96% ,Th2占 1 5 .66%± 3.95 % ;CD8+ T淋巴细胞中 Tc1占 1 6.2 2 %± 1 .78% ,Tc2占 9.2 0 %± 1 .96% ;而 1 2例妊高征孕妇的 Th1、Tc1百分比例明显高于正常人 ( P<0 .0 1 ) ,Th2的比例显著低于正常人( P<0 .0 1 )。 结论 双抗体标记流式细胞仪检测法可以客观、准确地检测淋巴细胞表达细胞因子的水平 ,判断 T淋巴细胞的极化状态。妊高征患者与正常人相比存在 T淋巴细胞的极化水平异常 。
Objective To explore a kind of accurate and sensitive method to detect intracellular cytokines, and to provide the method for research on relationship between profile of T lymphocytes and pathogenesis of some disorders. Methods In this report, blood was cultured in RPMI 1640 medium with a combination of PMA and ionomycin to reveal the cytokine synthetic potential of each cell, together with monensin to increase the sensitivity by retaining cytokines within the cell to detectable level, directly conjugated anti CD4/CD8 antibodies and anticytokines(IFNγ,IL 4) antibodies were used to label correspond antigens respectively,flow cytometry was employed to determine the expression of cytokines in CD4/CD8 T lymphocytes (namely,Th1/Th2 and Tc1/Tc2) . Results Assessment of IFNγ and IL 4 production within the CD4/CD8 lymphocytes from 18 normal persons and 12 patients with pregnancy induced hypertension (PIH) showed that more CD4/CD8 lymphocytes from patients with PIH produced IFNγ, however,less CD4 + lymphocytes secreted IL 4 than that of the control group (P<0.01). Conclusion This technique could be useful to determine whether cytokine assays are of value in the investigation of immune disorders. There is an abnormal profile of T lymphocytes from patients with PIH.
出处
《临床输血与检验》
CAS
2002年第1期3-6,共4页
Journal of Clinical Transfusion and Laboratory Medicine
基金
安徽省"九五"攻关项目 ( 96 130 0 7)资助
