人外周血白介素-22产生细胞在不同淋巴细胞亚群中的比例

目的:采用流式细胞术(flow cytom etry,FCM)检测人外周血不同淋巴细胞亚群中白介素-22(IL-22)产生细胞的比例。方法:健康人外周全血用肝素抗凝管采集后与相同体积RPM I 1640培养液混匀,加入佛波酯和离子霉素,于37℃、5% CO₂静置培养2 h,再加入莫能霉素后继续培养4 h收集细胞。用荧光素标记单抗进行细胞表面抗原和胞内细胞因子染色,用FCM检测表达IL-22、IL-17的细胞在不同淋巴细胞亚群中的比例。结果:IL-22产生细胞在CD3+T细胞、CD4+T细胞、CD8+T细胞、自然杀伤(NK)细胞和B细胞的表达比例分别为1.83%、2.12%、0.67%、1.96%和1.09%,CD4+T细胞中的比例明显高于CD8+T细胞中的比例(P<0.01)。另外,在CD3+T细胞、CD4+T细胞和CD8+T细胞中,均检测到共表达IL-22和IL-17的细胞,差异有统计学意义(P<0.01)。结论:用FCM检测IL-22产生细胞在CD4+T细胞、CD8+T细胞、NK细胞和B细胞均有表达,在CD4+T细胞的比例明显高于CD8+T细胞。

Distribulion of interleukin-22 producing cells in lymphocyte subpopulations from peripheral blood

Objective：To establish a method for detection of interleukin-22 producing cells in lymphocyte subpopulations from peripheral blood by flow cytometry.Methods：Peripheral blood samples were mixed with same volume of RPMI 1640 medium and stimulated with PMA and ionomycin for 6 hours in the presence of monensin at last 4 hours at 37 ℃,5% CO2.The stimulated cells were stained for surface molecules or intracellular cytokines with fluorochrome-conjugated mAbs.The proportions of IL-22 producing cells and IL-17 producing cells among different lymphocyte subsets were detected by flow cytometry.Results：The proportions of IL-22 producing cells among CD3＋T cells,CD4＋T cells,CD8＋T cells,NK cells and B lymphocytes were 1.83%,2.12%,0.67%,1.96% and 1.09%,respectively.The amount of IL-22 producing cells among CD4＋T cells was significantly higher than that among CD8＋T cells（P0.01）.In addition,IL-22 and IL-17 co-expressing cells were also detected in CD3＋T cells,CD4＋T cells,and CD8＋T cells.Conclusions：By using flow cytometry technique,the IL-22 producing cells exist among all CD4＋T cells,CD8＋T cells,NK cells and B lymphocytes.The proportion of IL-22 producing cells in CD4＋T cells is higher than that in CD8＋T cells.