梁友宝, 梁华, 李柏青. 白介素-9产生细胞在不同淋巴细胞亚群中的比例[J]. 蚌埠医学院学报, 2011, 36(5): 437-440.
    引用本文: 梁友宝, 梁华, 李柏青. 白介素-9产生细胞在不同淋巴细胞亚群中的比例[J]. 蚌埠医学院学报, 2011, 36(5): 437-440.
    LIANG You-bao, IANG Hua, LI Bai-qing. Distribution of interleukin-9 producing cells in different lymphocyte subpopulations detected[J]. Journal of Bengbu Medical College, 2011, 36(5): 437-440.
    Citation: LIANG You-bao, IANG Hua, LI Bai-qing. Distribution of interleukin-9 producing cells in different lymphocyte subpopulations detected[J]. Journal of Bengbu Medical College, 2011, 36(5): 437-440.

    白介素-9产生细胞在不同淋巴细胞亚群中的比例

    Distribution of interleukin-9 producing cells in different lymphocyte subpopulations detected

    • 摘要: 目的:应用流式细胞术测定胞内细胞因子,检测不同T淋巴细胞亚群和其他淋巴细胞亚群中白介素-9(IL-9)产生细胞的比例。方法:23名健康成年人外周抗凝血加等量1640培养液后,加佛波酯、离子霉素及莫能霉素培养6 h。收集细胞后,先用不同荧光素标记的抗人CD3、抗人CD8和抗人T细胞抗原受体γδ(TCRγδ)单抗,或抗人CD19和抗CD56单抗作细胞表面染色,再经过溶血,固定和透膜处理后,加抗胞内细胞因子的抗IL-9和IL-17的荧光抗体作胞内染色,用流式细胞仪检测和分析IL-9产生细胞在不同T淋巴细胞亚群和其他淋巴细胞亚群中的分布频率。结果:IL-9产生细胞在总T细胞(CD3+T)、CD4+T细胞、CD8+T细胞和γδT细胞中的百分比分别为0.35±0.31、0.22±0.13、0.31±0.29和1.07±1.05。γδT细胞中的IL-9产生细胞明显高于其他淋巴细胞亚群(P<0.01)。同时也发现自然杀伤细胞(NK细胞)中也有少量IL-9产生细胞。另外还发现,共表达IL-9和IL-17的细胞在CD3+T及其亚群CD4+T和CD8+T细胞中的比例很低(0.03%~0.06%),但在γδT细胞中的比例可达(0.28±0.34)%,差异有统计学意义(P<0.01)。结论:用流式细胞术测定单细胞水平胞内细胞因子的技术发现,不仅CD4+T细胞中,而且CD8+T细胞和γδT细胞以及NK细胞中均存在产生IL-9的细胞。

       

      Abstract: Objective: To detect the proportions of interleukin-9 producing cells in different lymphocyte subpopulations of peripheral blood of healthy subjects by intracellular cytokine measuring technique using flow cytometry.Methods: Twenty-three peripheral blood of healthy adults was stimulated by PMA and ionomycin,followed adding monensin for total 6 hours.The stimulated cells were stained with fluorescent antibodies,anti-CD3,anti-CD8,anti-TCRγδ,anti-CD19,and anti-CD56,followed hemolysing,fixed and membrane permeabilization,and stained intracellular cytokine with fluorescent anti-IL-9,anti-IL-17 antibodies.The proportion of IL-9 producing cells in different T cell subsets and other lymphocyte populations were detected and analyzed by Flow cytometry.Results: IL-9 producing cells were detected among all lymphocyte subpopulations,the percentages of which in total T cells(CD3+T),CD4+ T cells,CD8+ T cells and γδ T cells were 0.35±0.31,0.22±0.13,0.31±0.29 and 1.07±1.05,respectively.The proportion of IL-9 producing γδ T cells was significantly the highest(P<0.01).There was also the small proportion of IL-9 producing cells in NK cells.In addition,few amount of IL-9 and IL-17 co-expressing cells were detected among CD3+ T cells and CD4+T and CD8+T cells,but the small proportion of which in γδ T cells.Conclusions: The frequency of IL-9 producing cells detected by flow cytometry indicates that there exist IL-9 producing cells not only in CD4+ T cells,but also in CD8+T cells,γδ T cells and NK cells.

       

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