摘要
探讨IL-21对人外周血γδT细胞抗肿瘤活性的影响。分离健康人外周血单个核细胞(PBMC),分别在有或无IL-21参与的条件下,加入到含异戊烯焦磷酸(isopentenyl pyrophosphate,IPP)、IL-2或IL-15的RPMI 1640完全培养基中诱导培养γδT细胞。在培养的第10天用台盼蓝染色计数细胞;采用流式细胞术检测各组γδT细胞的纯度及γδT细胞穿孔素、颗粒酶B和CD107a的表达;用CCK-8试剂盒检测γδT细胞对K562细胞的体外杀伤效应。结果显示,不同细胞因子组诱导10d后γδT细胞纯度均达到70%以上,IL-2+IL-21、IL-15+IL-21和IL-2+IL-15+IL-21组与IL-2、IL-15组相比,细胞纯度和扩增倍数没有显著性变化;IL-2+IL-21、IL-15+IL-21和IL-2+IL-15+IL-21组γδT细胞穿孔素、颗粒酶B和CD107a的表达及对K562细胞的杀伤活性均显著高于IL-2、IL-15单独组。以上结果表明,IL-21可通过上调γδT细胞穿孔素和颗粒酶B的表达增强其抗肿瘤活性。
To investigate the enhancing effect of IL-21 on antitumor activity of γδT cells in vitro,γδT cells were generated in vitro by stimulating peripheral blood mononuclear cells of healthy donors with isopentenyl pyrophosphate（IPP） and IL-2 or IL-15 with or without IL-21（20 ng/μl） respectively.γδT cells number of each group were countered by trypan blue staining after 10 d culture.The purity of γδT cells and their perforin,granzyme B and CD107a expression were verified by flow cytometer.The cytotoxic activity of γδT cells against K562 cells were analyzed by CCK-8 kit.The experimental results showed that γδT cells purity of different cytokine groups were all exceeded 70% after 10 d culture.Comparing with IL-2 alone or IL-15 alone group,expansion folds and purity of γδT cells induced by cytokine combination groups,that is IL-2＋IL-21、IL-15＋IL-21 and IL-2＋IL-15＋IL-21 groups,had no significant differnce.However,perforin,granzyme B and CD107a expression of γδT cells of IL-2＋IL-21、IL-15＋IL-21 and IL-2＋IL-15＋IL-21 groups were significantly higher than that of the IL-2 alone or IL-15 alone group.Moreover,the γδT cell cytotoxicity to K562 cells of combination groups were significantly higher than IL-2 or IL-15 alone group.These results indicate that IL-21 could enhance γδT cell-mediated antitumor activity by up-regulation of perforin and granzyme B expression.
出处
《现代免疫学》
CAS
CSCD
北大核心
2012年第3期226-229,共4页
Current Immunology
基金
南京军区医学科技创新经费资助项目(NO:08MA039)
