体外检测人Th17细胞频率的方法

目的探讨不同刺激对象、刺激剂量、刺激时间、蛋白质运送抑制剂对体外检测人Th17细胞频率的影响。方法用不同浓度的佛波酯（PMA，终浓度25、50、150、300、500、750、1000ng／mL）与离子霉素（Ionomycin）对健康人全血和外周血单个核细胞（PBMC）进行刺激，1h后加蛋白质转运抑制剂布雷杆氏菌素A（BFA）或莫能菌素（Monensin），分别培养3h、6h、9h，收集细胞，以荧光素-mAb标记后，用流式细胞术分析Th17频率。结果全血刺激好于PBMC对Th17频率的检测效果（P〈O．05），PMA300ng／mL刺激全血时Th17频率最高，BFA或Monensin阻断IL-17分泌效果相同但溶血有差异，刺激6h最适宜。结论PMA300ng／mL＋Ionomycin刺激全血1h后加Monensin2μmol／L继续培养6h，是检测人Th17细胞频率最适宜的方法。

In vitro detection of the frequency of human Thl7 cells

Objective To investigate the effect of different stimulated objects, different stimulating concentrations, different protein transportation inhibitors on the frequency of human Th17 cells. Methods Whole blood or peripheral blood mononuclear cells (PBMC) were stimulated by PMA at different concentrations(25, 50, 150, 300, 500, 750, and 1 000ng/mL). Ionomycin, different protein transportation inhibitors (BFA or Monensin) were added after 1h and then Th17 cells were detected after different stimulating time po...