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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 To evaluate the distribution characteristics of Th1 / Th2 subsets in cytokine-induced killer cells (CIK) by magnetic bead sorting system and intracellular staining, and to explore the value of intracellular staining. Methods CIK was amplified in large scale in vitro, and the human leukocyte differentiation antigen (CD) 4 + T cell subsets in CIK were enriched by magnetic bead separation system. The distribution of Th1 / Th2 cell subsets was analyzed by intracellular staining. Results The purity of CD4 + CIK cells enriched by magnetic bead separation was as high as 96%, which was significantly different from that of peripheral blood mononuclear cells (PBMC). Th1 (IFN-γ + IL-4) γ + IL-4 +) increased from (1.18 ± 0.94)% and (0.19 ± 0.16)% to (33.93 ± 6.38)% and ± 8.23%, respectively (P <0.01), while the Th2 (IFN-γIL-4 +) subpopulation decreased from (17.56 ± 16.71)% to (10.21 ± 7.05)% , But the difference was not statistically significant (P> 0.05). Conclusions This study suggests that CIK has obvious “Th1 predominance”. The combination of magnetic bead sorting system and intracellular staining in determining the distribution of Th1 / Th2 cell subsets is simple, rapid, stable and suitable for clinical application.