【摘要】 目的探讨丁基羟基茴香醚(BHA)在体外诱导扩增CD4~+中央记忆性T细胞(TCM)中的作用,提高过继免疫疗法的有效性。方法将健康人外周血单个核细胞使用密度梯度离心的方法分离,磁珠分选得到CD4~+T淋巴细胞,将细胞分为实验组和对照组,实验组加入不同浓度的BHA(10、20、40μmol/L)),对照组加入等体积的PBS,经过细胞培养使用流式细胞仪对其TCM细胞亚群的分化扩增进行检测。结果流式结果显示,实验组的CD4~+TCM中有扩增活性的细胞比例明显增加,其中对照组的比例为73.2%,低剂量组的比例为78.8%,中剂量组的比例为87.7%,高剂量组为89.9%,并有剂量依赖效应。对照组的CD45RA-CD62L+CCR7+TCM细胞占总CD4~+T细胞的比例为10.60%;BHA浓度为10、20、40μmol/L时比例分别为23.52%、32.39%、28.41%。对体外培养15 d后的细胞进行了凋亡检测,结果显示,与对照组比较,实验组中PI+Annexin V+发生凋亡的细胞从27.1%下降到了12.5%。结论在CD4~+T细胞的体外扩增实验中,BHA可以显著提高TCM细胞亚群的比例,在CD4~+T的体外培养过程中,BHA可以显著抑制细胞的凋亡。更多还原

【Abstract】 Objective To study the effect of Butylated hydroxyanisole(BHA), a kind of small-molecule compound,on the in vitro expansion of CD4~+central memory T cells(TCM). Methods Human CD4~+Tlymphocytes were isolated from the peripheral blood mononuclear cells(PBMC) from several healthy donors. Then, the CD4~+Tlymphocytes were divided into two groups(control group and experimental group) and stimulated by anti-CD3,anti-CD28 and BHA. The expansion of CD4~+TCM was detected by flow cytometry. Results Data indicated that the proportion of CD4~+TCM with high expansion activity increased significantly in the experimental group. The proportion of TCM with high expansion activity was 73.2% in control group,compared with a proportion of 78.8% in low dosage group, 87.7% in middle dosage group and 89.9% in high dosage group. The results indicated that BHA can increase the proportion of expanding TCM in a dose-dependent manner. Moreover, the proportions of CD45RA-CD62L+CCR7+TCM in total CD4~+T cells were10.60%,23.52%,32.39%,28.41% when the concentration of BHA were 0,10, 20, 40 μmol / L respectively. Last but not least, compared with the TCM apoptosis rate of 27.1% in control group, the TCM apoptosis rate was 12.5% in BHA group, which suggested that BHA can also dramatically decease the cell apoptosis of CD4~+T lymphocytes. Conclusion BHA can increase the proportion of TCM in CD4~+Tlymphocytes; thereby generating a new method to improve the efficiency of adoptive cell immunotherapy.更多还原