CpGODN2216对白血病缓解期患者外周血单个核细胞的活化能力及活化的细胞对K562细胞的杀伤作用

本研究探讨CpGODN2216对白血病缓解期患者外周血单个核细胞（peripheral blood mononuclear cells，PBMNC）的活化能力以及CpGODN2216活化的PBMNC对白血病细胞K562的杀伤作用。取白血病缓解期患者外周血，以淋巴细胞分离液（Ficoll）分离PBMNC，用含10％胎牛血清的RPMI1640培养液调整PBMNC浓度为2．0×10^6／ml。对照组加入生理盐水（NS），实验组加入CpGODN2216，培养1周，提取上清，用ELISA检测上清中IFN-γ、IL-12、IL-4、IL-10的水平。取白血病缓解期患者外周血，分离PBMNC，对照组加入生理盐水，实验组加入CpGODN2216，培养48小时，用流式细胞仪检测PBMNC中淋巴细胞的Th1／Th2，Tc1／Tc2细胞比例；流式细胞术检测活化的PBMNC对肿瘤细胞K562的杀伤能力。结果表明：实验组上清中IFN-γ、IL-12水平与对照纽相比有显著差异（P〈0．01）；IL-4、IL-10水平与对照组相比无明显差异（P〉0．05）。CpGODN2216能刺激白血病缓解期患者PBMNC向Th1／Tc1转化，与对照组相比有显著差异（P〈0．01），但对Th2／Tc2无明显的刺激作用（P〉0．05）。CpGODN2216活化的PBMNC对K562细胞的杀伤能力明显高于对照组（P〈0．01）。结论：CpGODN2216可在体外诱导白血病缓解期患者PBMNC分泌Th1型细胞因子，诱导PBMNC向Th1方向转化，并强烈地活化CD8^＋T细胞应答。CpGODN2216活化的PBMNC对白血病细胞K562具有较强的杀伤作用.

Activation Ability of CpG ODNs 2216 on PBMNCs from Leukemia Patients in Remission and Killing Effect of Activated PBMNCs on 1K562 Cells

The aim of this study was to investigate the activation ability of CpG oligodeoxynucleotide （ CpG ODN） 2216 on the peripheral blood mononuclear cells（PBMNCs） from leukemia patients in remission and the killing effect of activated PBMNCs on K562 cells. PBMNCs obtained from leukemia patients in remission were incubated with CpG ODN 2216. In control group, PBMNCs were incubated with normal saline （NS）. The concentrations of cytokines （ IFN-γ, interleukin-12, interleukin-4, interleukin-10） in culture supernatant of PBMNCs from leukemia patients in remission were analyzed by using ELISA kits. The percentages of Thl ,Tc1 ,Th2 ,Tc2 cells and killed K562 cells were detected by flow cytometry. The results showed that as compared with control group, CpG ODN 2216 induced higher concentrations production of IFN-γ, IL-12 in supernatant（p 〈0.01 ）. There were no differences in IL-4,IL-10 in supernatant as compared with control group （p 〉0.05）. The percentages of Th1 and Tcl cells increased significantly after culture with CpG ODN 2216 as compared with control group（p 〈 0.05 ）. There was no difference between the percentages of Th2 and Tc2 cells in stimulated group and control group. The killing effect of PBMNCs on K562 cells was significantly different between the stimulated group and control group（p 〈 0.05）. It is concluded that CpG ODNs 2216 can induce strong Th1-like immune activation, with the secretion of type-I cytokine and activation of strong CD8^ ＋ T-cell responses. PBMNCs activated by CpG ODNs can more strongly kill k562 cells in vitro.