摘要
目的 :观察CIK细胞中CD4 + T细胞亚群抗肿瘤免疫活性。方法 :体外大规模扩增CIK细胞 ,利用磁珠分离系统富集纯化CIK细胞中的CD4 + T细胞亚群。采用胞内染色法分析其中Th1/Th2的比例变化 ;LDH法和荧光染色法比较 4h和 2 0h其对raji细胞的杀伤率和raji凋亡。 结果 :经磁珠分离法富集的CD4 + CIK细胞纯度高达 96 % ,其中Th1/Th2的分布较PBMC有显著的改变 :Th1亚群、Th0亚群明显升高 ,Th2亚群无显著变化。CD4 + CIK细胞虽然不能在 4h之内溶解raji细胞 ,但可在 2 0h时产生同CD4 -CIK细胞同样强大的杀伤活性 ,荧光染色可见其在 4h之内诱导raji出现早期凋亡的迹象。 结论 :本研究提示CD4 + CIK细胞具有明显的“Th1优势”可以调节宿主免疫细胞活性 ;同时CD4 + CIK细胞可通过诱导肿瘤细胞凋亡实现对肿瘤的抑制和杀伤。
Objective: To observe antitumor immunity of CD4+ T cells subset in CIKs. Methods: After large scale of amplification in vitro, CD4+ T cells subset in CIKs was isolated by magnetic beads separation columns. Distribution of Th1/ Th2 in CD4+ T cells subset in CIKs was analysized by intracellular cytokine staining. Cytotoxicity of purified CD4+ T cells subset in CIKs against raji cells and apoptosis of raji cells after 4 h and 20 h coculture were determined by LDH method and fluorescent staining method. Results: Purity of enriched CD4+ T cells subset in CIKs reached 96%. Comparing with PBMCs, significant increase in Th1 subset and Th0 subset were observed but no statistical differences were found in Th2 subset. Few raji cells were lysed by CD4+ T cells subset in CIKs after 4 h co-incubation. But after 20 h co-incubation, the same effective lysis of raji cells as CD4- T cells subset was obtained in CD4+ T cells subset in CIKs. Fluorescent staining showed that CD4+ T cells subset in CIKs induced apoptosis of raji after 4 h coculture. Conclusion: The present study suggested that CD4+ T cells in CIKs were not only regulatory cells capable of modulating host immune system, but also immune effectors capable of inducing apoptosis in tumor cells.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2003年第2期126-129,共4页
Chinese Journal of Cancer Biotherapy
基金
天津科委自然科学基金资助项目 (项目编号02 3 6110 11)
