磁珠分选系统和胞内染色法联合分析细胞因子诱导杀伤细胞中Th1/Th2细胞亚群分布特点  

Analysis of Th1/Th2 subsets distribution in CIKs by intracellular cytokine staining method combined with magnetic beads separation system

作　　者：任秀宝[1] 于津浦[1] 刘虹[1] 张澎[1] 郝希山[1] 

机构地区：[1]天津医科大学附属肿瘤医院免疫室,300060

出　　处：《中华检验医学杂志》2005年第5期486-488,共3页Chinese Journal of Laboratory Medicine

基　　金：天津科委自然科学基金资助项目(023611011)

摘　　要：目的　评价以磁珠分选系统和胞内染色法联合分析细胞因子诱导杀伤细胞(CIK)中Th1 /Th2细胞亚群分布特点,探讨胞内染色法的应用价值。方法　体外大规模扩增CIK,利用磁珠分离系统富集纯化CIK中的人白细胞分化抗原(CD)4+T细胞亚群。用胞内染色法分析其Th1 /Th2细胞亚群的分布特点。结果　经磁珠分离法富集的CD4+CIK细胞纯度高达96%,它与外周血单个核细胞(PBMC)相比变化显著,其中Th1 (IFN -γ+IL- 4 )亚群、Th0 (IFN -γ+IL- 4+ )亚群分别从( 1 .18±0. 94)%和( 0. 19 ±0. 16 )%升至( 33 .93 ±6 .38 )%和( 23. 07 ±8. 23 )% (P< 0 .01 ),而Th2(IFN -γIL- 4+ )亚群虽然从( 17 .56±16. 71 )%降至( 10 .21±7. 05 )%,但差异无统计学意义(P>0. 05)。结论　本试验提示CIK具有明显的“Th1优势”;磁珠分选系统和胞内染色法联合测定Th1 /Th2细胞亚群分布的方法简便快捷,结果稳定,适合临床应用。Objective By analyzing the distribution of Th1/Th2 subsets in CD4 + CIKs to identify the clinical applicable value of intracellular staining method combined with magnetic beads separation system.Methods After amplification, CD4 + CIKs were enriched by MACS magnetic beads separation system.The characteristic of Th1/ Th2 subsets distribution was analyzed by intracellular cytokine staining methods.Results Purity of enriched CD4 + CIKs reached 96%.Compared with PBMCs, significant changes were observed in Th1 subset and Th0 subset of CIKs, which increased from (1.18±0.94)% and (0.19±0.16)% to(33.93±6.38)% and (23.07±8.23)% respectively, with P value <0.01 both. But no statistical significance was identified in Th2 subset which dropped from (17.56±16.71)% to(10.21±7.05)%.Conclusion This study suggested that “Th1 dominance” exist in CIKs.As a valuable method to study Th1/Th2 subsets distribution, intracellular staining method combined with magnetic beads separation system gives swift and repeatable result that suit for clinical usage.

关 键 词：TH1/TH2细胞亚群 细胞因子诱导杀伤细胞 联合分析 染色法 胞内 磁珠 分布 分选 人白细胞分化抗原 外周血单个核细胞 IFN-γ T细胞亚群 CD4^+ CIK细胞 应用价值 富集纯化 分离系统 联合测定 临床应用 分离法 统计学 

分 类 号：R73-3[医药卫生—肿瘤]