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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 effect of IL-21 on the anti-tumor activity of human peripheral blood γδT cells. Peripheral blood mononuclear cells (PBMC) from healthy individuals were isolated and added to RPMI 1640 containing isopentenyl pyrophosphate (IPP), IL-2 or IL-15 with and without the intervention of IL-21 Induction of γδT cells in complete medium. The cells were counted by trypan blue staining on day 10 of culture. The purity of γδT cells and the expression of perforin, granzyme B and CD107a of γδT cells were detected by flow cytometry. The expression of γδT cells was detected by CCK-8 kit K562 cells in vitro killing effect. The results showed that the purity ofγδT cells reached more than 70% after induced by different cytokines for 10 days. IL-2 + IL-21, IL-15 + IL-21 and IL-2 + IL-15 + IL- IL-2 + IL-21, IL-15 + IL-21 and IL-2 + IL-15 + IL-21 group compared with IL- The expression of perforin, granzyme B and CD107a and the cytotoxicity against K562 cells were significantly higher than that of IL-2 and IL-15 alone groups. The above results show that IL-21 can enhance its anti-tumor activity by up-regulating the expression of perforin and granzyme B of γδT cells.