急性髓细胞性白血病细胞培养上清对CD4+ 及CD8+ T细胞增殖及凋亡.doc

急性髓细胞性白血病细胞培养上清对CD4+ 及CD8+ T细胞增殖及凋亡 　 作者：王兴兵，刘隽，贺艳丽，谷俊侠，郑金娥，姚军霞，杨晶，李小青，黄士昂 【摘要】 为了研究不同的急性髓细胞性白血病（AML）细胞株培养上清对CD4+和CD8+ T细胞增殖以及凋亡的影响，探讨AML的免疫抑制的形成机制，将3种AML细胞株（HL-60、NB4和U937）培养上清和普通培养液按不同比例混合，用于培养CFSE染色的淋巴细胞。抗CD3抗体和抗CD28抗体刺激3天后，标记7AAD和相应荧光抗体。利用流式细胞术检测不同T细胞亚群CFSE荧光强度和7AAD表达率的变化，分析其增殖和凋亡变化情况。结果表明： 3种AML细胞株中有2种（HL-60和NB4）培养上清可显著抑制CD4+ T细胞和CD8+ T细胞的增殖，并随浓度增加而增强。同样，HL-60和NB4细胞株培养上清亦可抑制刺激后的CD4+ T细胞的凋亡，但对刺激后的CD8+ T细胞的凋亡并无明显影响。相反，HL-60和NB4细胞株培养上清可显著增加增殖的CD8+ T细胞的凋亡。结论： AML细胞株培养上清液对CD4+ T细胞和CD8+ T细胞增殖的抑制以及对增殖的CD8+ T细胞凋亡的诱导作用，可能是AML患者免疫功能缺陷的机制之一。 【关键词】 急性髓细胞性白血病 　　Effects of Acute Myeloid Leukemia Cell Supernatant on the Proli-feration and Apoptosis of CD4+ and CD8+ T Cell Subsets 　　　Abstract To study the effects of supernatant derived from acute myeloid leukemia (AML) cell lines on proliferation and apoptosis of CD4+ and CD8+ T cell subsets and to investigate the mechanism by which AML escapes from immune recognition，lymphocytes were labeled with CFSE and were stimulated with anti-CD3 and anti-CD28 in presence or absence of supernatants from three AML cell lines (HL-60，NB4，U937). After culture，cell suspensions were labeled with 7AAD and CD4 PE (or CD8 PE). Cells were then detected by flow cytometry and their proliferation and apoptosis were analyzed. The results showed that supernatants from two of three cell lines (HL-60 and NB4) inhibited the proliferation of CD4+ and CD8+ T cells，and the degree of inhibition showed a dose-dependent way. Similarly，the apoptosis of stimulated CD4+ T cells was inhibited，but stimulated CD8+ T cells remained unaffected by supernatant from HL-60 and NB4. In contrary，the apoptosis of proliferative CD8+ T cells were increased significantly by HL-60 and NB4 supernatant. It is concluded that soluble factors derived from AML cell lines inhibit the proliferation of CD4+ and CD8+ T cells and induce the apoptosis of proliferative CD8+ T cells，that may be one of the mechanisms by which the immunity was suppressed. 　　Key words acute myeloid leukemia; CD4