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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。
To investigate the effect of different culture supernatants of AML on the proliferation and apoptosis of CD4 + and CD8 + T cells, the mechanism of AML immunosuppression was studied. Three AML cell lines (HL-60 , NB4 and U937) The culture supernatant and normal culture medium were mixed in different proportions and used to culture CFSE-stained lymphocytes. After 3 days of stimulation with anti-CD3 and anti-CD28 antibodies, 7AAD and the corresponding fluorescent antibody were labeled. Flow cytometry was used to detect the changes of the fluorescence intensity and the expression rate of 7AAD in different T cell subsets. The changes of proliferation and apoptosis were analyzed. The results showed that two of the three AML cell lines (HL-60 and NB4) could significantly inhibit the proliferation of CD4 + T cells and CD8 + T cells and increase with increasing concentration. Similarly, the supernatant of HL-60 and NB4 cell lines also inhibited the apoptosis of stimulated CD4 + T cells, but had no significant effect on the apoptosis of stimulated CD8 + T cells. In contrast, the supernatant of HL-60 and NB4 cell lines significantly increased the apoptosis of proliferating CD8 + T cells. Conclusion: The inhibition of proliferation of CD4 + T cells and CD8 + T cells by AML cell culture supernatant and the induction of apoptosis of proliferating CD8 + T cells may be one of the mechanisms of immune dysfunction in AML patients.