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

摘要

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 (P ＜0. 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.%目的:采用流式细胞术(flow cytometry,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RPMI 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业群中的比例.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.