非小细胞肺癌患者外周血髓样抑制细胞比例及其对CD8^+T细胞抑制作用的测定  

The change of proportion and inhibition on CD8^+T lymphocytes of myeloid-derived suppressor cells in peripheral blood from patients with non-small cell lung cancer

作　　者：符一宁[1] 王士勇[1] 马克骥 刘畅[1] 焦雪[1] 刘迪杰[1] 王佳玲[1] 张晖[1] 张哲[1] 杜微丽[1] 张璐[1] 武秀艳[1] 何英[1] 梁春艳[1] 刘殊[1] 

机构地区：[1]中国医科大学附属第四医院生物治疗科,辽宁沈阳110032 [2]营口市中心医院普外二科,辽宁营口115000

出　　处：《现代肿瘤医学》2013年第10期2224-2227,共4页Journal of Modern Oncology

基　　金：国家自然科学基金课题(编号:81200573);辽宁省自然科学基金资助项目(编号:201102296)

摘　　要：目的:探讨非小细胞肺癌(NSCLC)患者外周血中髓样抑制细胞(MDSCs)的比例及其对CD8+T细胞的抑制作用。方法:密度梯度离心法分离30例NSCLC患者及15例健康对照者外周血单个核细胞(PBMC),流式抗体染色后经流式细胞仪检测MDSCs中CD11b、CD14和CD33的表达;磁珠分选法分离、纯化肿瘤患者外周血中的MDSCs及健康者外周血中的CD8+T细胞,1∶1共培养72小时,用ELISA法测定24、48、72小时细胞培养上清液的INF-γ水平。结果:30例进展期NSCLC患者外周血中MDSCs比例为(25.1±16.8)%,明显高于健康对照组(8.2±3.6)%,两者差异有统计学意义(P<0.001);未观察到MDSCs比例与患者年龄、性别、分期及病理类型有关(P>0.05);ELISA法显示:与CD8+T细胞单独培养组相比,MDSCs与CD8+T细胞组共培养24、48、72小时的培养上清IFN-γ水平由(201.3±14.57)ng/ml逐渐下降为(163.33±7.77)ng/ml、(132.0±6.9)ng/ml和(79.67±7.09)ng/ml,明显降低(P<0.05)。结论:进展期NSCLC患者外周血中MDSCs的比例较健康者明显升高,且对CD8+T细胞有抑制作用,MDSCs增多可能是NSCLC患者发生免疫抑制的重要原因之一。Objective： To investigate the change of proportion and suppressive on CD8＋ T lymphocytes of myeloid -derived suppressor cells （MDSCs） in peripheral blood from patients with non -small cell lung cancer（ NSCLC ）. Methods： The proportion of MDSCs in peripheral blood mononuclear cells （ PBMC ） was determined in 30 patients with NSCLC and 15 healthy control subjects on a Ficoll - Paque density gradient. Flow cytometry was used to detect the expression of CD1 l b, CD14 and CD33. To culture CD1 l b ＋/CD14- cells from NSCLC, CD8＋ lymphocytes from control subjects and co -culture between CD1 l b＋/CD14- cells and CD8 ＋ lymphocytes with the ratio of 1：1 for 72 hours after magnetic bead isolation. Then examine the INF - γ fluctuations of co - euhured cell supernatant on 24,48, 72 hours by ELISA. Results：The proportion of MDSCs of NSCLC patients was （25.1 ± 16.8 ）%, significantly higher than healthy control subjects （8.2 ±3.6） % （P 〈0.001 ）. There were no significant differences in the subpopulation of cells between patients with different age, gender, stage and type of histology（ P 〉 0.05 ）. Compared with CD8 ＋ lym- phocytes, the INF -γ fluctuations of co -culture cell supernatant on 24,48,72 hours by ELISA was gradually reduced from （201.3± 14.57 ） ng/ml to （ 163.33 ± 7.77 ） ng/ml, （ 132.0 ± 6.9 ） ng/ml and （79.67 ± 7.09 ） ng/ml （ P 〈 0.05 ）. Conclusion： The proportion of MDSCs in PBMC of advanced - stage NSCLC patients was significantly higher than healthy control subjects, and have the suppressive effect of the cells on CD8＋ T lymphocytes, the increase of MDSCs may be one of important reasons of NSCLC patients for immunosuppression.

关 键 词：髓样抑制细胞 非小细胞肺癌 免疫抑制 磁式分选 

分 类 号：R734.2[医药卫生—肿瘤]