不同IL-15基因转染对NCI-H446细胞诱导PBMC增殖和杀伤肿瘤细胞的影响

目的 研究不同IL-15基因转染对NCI-H446细胞诱导外周血单个核细胞(PBMC)增殖和杀伤肿瘤细胞的影响.方法 野生型NCI-H446细胞(Cw)和被3种IL-15基因分别转染的3种NCI-H446细胞(Cmp:被IL-15成熟肽基因转染;Cp:被原型IL-15基因转染;Csp:被信号肽换为IL-2信号肽的改型IL-15基因转染),用丝裂霉素(M)处理后(分别名为MCw、MCmp、MCp和MCsp)作为刺激细胞刺激健康志愿者的PBMC.对这些被刺激的PBMC,分别名为MCw-PBMC、MCmp-PBMC、MCp-PBMC和MC-sp-PBMC.台盼蓝拒染法计数细胞数,流式细胞术测定CD4 细胞和CD8 细胞百分率.在MCmp-PBMC,MCp-PBMC和MCsp-PBMC中,选择其细胞数和CD4 细胞和/或CD8 细胞百分率统计学上明显高于MCw-PBMC的作为效应细胞,MTT法测定它们对Cw的杀伤.结果 与MCw-PBMC相比,MCp-PBMC在细胞数、CD4 细胞和CD8 细胞百分率及对Cw的杀伤上,均显著提高(P＜0.05).结论 原型IL-15基因转染能提高NCI-H446细胞诱导PBMC增殖和杀伤野生型NCI-H446细胞的能力.

Effects of different IL-15 gene transfections into NCI-H446 cells on inducing PBMC to proliferate and to kill tumor cells

Objective To investigate the effects of different IL-15 gene transfections into NCI-H446 on inducing peripheral blood mononuclear cells (PBMC) to proliferate and to kill tumor cells. Methods After the wide type NCI-H446 cells (Cw) and the three kinds of NCI-H446 cells transfected by three kinds of IL-15 genes respectively (Cmp transfected by the IL-15 maturation peptide gene, Cp transfected by prototypic IL-15 gene, and Csp transfected by modified IL-15 gene whose signal peptide was replaced with the IL-2 signal peptide) were treated by mitomycin (M), the cells (named MCw, MCmp, MCp, and MCsp, respectively) were used as stimulator cells to stimulate PBMC from healthy volunteers. For these stimulated PBMC, named MCw-PBMC, MCmp-PBMC, MCp-PBMC, and MCsp-PBMC respectively, the number of cell was counted by trypan blue exclusion staining and the percentages of CD4 cells and CD8 cells were detected by FCM. The PBMC which had higher cell proliferation and percentages of CD4 cells and/or CD8 cells than MCw-PBMC were selected and treated as effector cells, and then the killing activity of the selected PBMC against Cw was measured by MTT. Results Compared with MCw-PBMC, the MCp-PBMC in the cell proliferation, the percentages of CD4 cells and CD8 cells, and the Cw-killing activity was significantly enhanced (P<0.05). Conclusion The prototypic IL-15 gene transfection could enhance the ability of NCI-H446 cells to induce PBMC to proliferate and to kill the wild type NCI-H446 cells.