摘要
目的:观察CIK细胞中CD4+T细胞中Th1/Th2亚群分布情况。方法:体外大规模扩增CIK细胞,磁珠法纯化其中CD4+T细胞。采用胞内染色、ELISA和荧光定量RT-PCR法检测培养前后Th1/Th2细胞亚群的比例、Th1/Th2类细胞因子的分泌量和基因表达的变化。结果:富集CD4+细胞纯度达96%,CIK中Th1亚群和Th0亚群的比例较PBMC显著升高(P<0.05),而Th2亚群无显著变化。CD4+CIK细胞中IL-2和IFN-γ等Th1类细胞因子释放量增高(P<0.01),而IL-10和TGF-"等Th2类细胞因子释放量降低(P<0.05)。但CD4+CIK细胞中除IFN-γ的mRNA显著升高外(P<0.05),IL-2和IL-4的mRNA变化不明显。结论:CIK细胞中的CD4+T细胞具有明显的Th1优势。
Objective： To observe the distribution of Th1/Th2 subsets in CD4^＋T cells of cytokineinduced killer （CIKs）. Methods： After large scale of amplification of CIKs, CD4^＋T cells subset was isolated by magnetic beads separation columns, and the proportion of IFN-γand IL-4 secreting cells were analyzed by intracellular cytokine staining. The cytokines of Th1 kind and Th2 kind, released by purified CD4^＋T ceils from the CIKs or PBMCs, were detected by ELISA and fluorescent real-time quantitive RT-PCR methods. Results： Purity of enriched CD4^＋T cells reached 96%. Comparing with PBMCs, a significant increase （P〈0.05） in Thl subset （IFN-γ^＋ IL-4^- cells） and Th0 subset （IFN-γ^＋ IL-4^＋ cells） was respectively observed while no statistical change （P〉0.05） was found in Th2 subset（IFN-γ^- IL-4^＋ cells）in CD4^＋ CIKs. Secretion of Th1 kind of cytokines, such as IL-2 and IFN-γ in CD4^＋T cells of CIKs enhanced significantly, and Th2 kind of cytokines, such as IL-10 and TGF-β decreased obviously （P〈0.05）. Except for IFN-γ no increase in mRNA synthesis of IL-2 or IL-4 was observed in CD4^=＋T cells in CIKs compared to PBMCs. Conclusion： This study indicates that a distinct “Th1 dominance” exists in CD4^＋T cells of CIKs.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2005年第14期795-797,共3页
Chinese Journal of Clinical Oncology
基金
天津市科委自然科学基金资助(编号:023611011)
