【摘要】 目的研究源于细菌 CpG 基序的寡核苷酸和含细菌短磷酸二酯骨架的寡核苷酸对K562/A02细胞来源的树突细胞(DC)的促成熟作用。方法联合应用细胞因子 rhGM-CSF 和 rhIL-4诱导 K562/A02细胞成为 DC。7 d 后以瑞特-姬姆萨染色观察细胞形态的变化,流式细胞术检测细胞免疫表型的变化,用同种混合淋巴细胞反应、细胞毒性 T 淋巴细胞(CTL)杀伤活性检测、IL-12和 IL-6的分泌实验评价 DC 的成熟程度。然后在 DC 中分别加入源于细菌 CpG 基序的寡核苷酸 CpG2006以及含细菌短磷酸二酯骨架的寡核苷酸 A-ODN 和 T-ODN,处理3 d 后再次检测该 DC 成熟度。结果K562/A02细胞可在细胞因子 rhGM-CSF 和 rhIL-4联合作用下分化成为 DC,免疫表型检测发现 CD83、HLA-DR 和 CD86分子的表达分别为(65.5±8.4)%、(32.0±4.3)%和(18.6±3.2)%,经 CpG2006作用后表达升高到(88.9±3.6)%、(53.9±3.2)%和(39.9±7.3)%;经 A-ODN 作用后升高到(97.0±5.3)%、(63.9±7,3)%和(40.2±7.4)%;经 T-ODN 作用后升高到(93.3±4.6)%、(58.3±5.6)%和(36.2±6.8)%,与作用前相比差异均有统计学意义。经 CpG2006、A-ODN 和 T-ODN 作用后具有典型DC 形态的细胞增多。上述3种寡核苷酸处理的 DC 均可刺激 T 细胞产生较强的增殖效应、诱导产生的 CTL 对靶细胞 K562/A02的杀伤率明显增强,IL-6和 IL-12分泌明显增高。结论源于细菌 CpG 基序的寡核苷酸以及含细菌短磷酸二酯骨架的寡核苷酸对白血病源性 DC 有促成熟作用。更多还原

【Abstract】 Objective To study the maturation effect of CpG2006 and phosphodiester oligonucleoti- des on leukemia-derived dendritic cells.Methods Leukemia cells K562/A02 were induced into dendritic cells by rhGM-CSF and rhIL-4.After 7 days induction,the cell-morphology was observed,the immunopheno- type of cells was detected by flow cytometry and the cell function was evaluated by allogeneic mixed lympho- cyte reactions,CTL responses and secretion of IL-12 and IL-6.Then a CpG oligonucleotide CpG2006,two synthetic bacterial phosphodiester oligonucleotides A-ODN and T-ODN were added to these leukemia-derived DCs.Three days later,the DCs were redetected by the above-mentioned methods.Results After induced by CpG2006,A-ODN or T-ODN,the leukemia-derived DCs with typical dendritic morphology were increased. The expressions of CD83,HLA-DR and CD86 were(65.5±8.4)%,(32.0±4.3)% and(18.6±3.2)% respectively in day 7 leukemia-derived DCs,raised to(88.9±3.6)% ,(53.9±3.2)% and(39.9± 7.3)% respectively after exposing CpG2006 for 3 days;increased to(97.0±5.3)% ,(63.9±7.3)% and (40.2±7.4)% respectively after treated by A-ODN;and further increased to(93.26±4.65)%,(58.3± 5.6)% and(36.2±6.8)% respectively after treated by T-ODN.These results was markedly different than unaffected cells did.These DCs induced by the above-mentioned three oligonucleotides could upregulate sig- nificantly the capacity for stimulating allogeneic T cells.They could also induce CTL to generate specific cyto- toxic activity against K562/A02 cells.The secretion of IL-6 and IL-12 was increased remarkably.Conclusion CpG2006,as well as two phosphodiester oligonucleotides can induce leukemia-derived DCs maturation.更多还原