目的： 探讨基于第一/第二刺激信号原理联合IL-15的的T淋巴细胞扩增技术（PersonGen TM ）和细胞因子诱导的杀伤（cytokine-induced killer，CIK）细胞培养扩增方案对T细胞扩增和活化的效果。方法：分离8名健康人外周血单个核细胞（peripheral blood mononuclear cell，PBMC），分别采用CIK细胞扩增技术（CIK组）、PersonGen TM 扩增技术（PersonGen TM 组）及两种技术的联合应用（C＋P组）刺激PBMC增殖，应用细胞计数法和流式细胞术比较T细胞的扩增效率及T细胞亚群比例，并以人白血病K562细胞及骨髓瘤RPMI 8226细胞为靶细胞检测扩增后T细胞的杀伤活性。结果：经过3周培养后，CIK、PersonGen TM 和C+P 3组T细胞扩增倍数分别为（254±56）、（863±218）和（793.3±395）倍。其中CD3 +细胞所占比例分别为（51.61±14.95）%、（99.21±0.79）%和（96.14±5.16）%； CD3 +CD4 +在CIK组细胞扩增了近5倍，C＋P组与PersonGen TM 组均扩增近160倍；CD3 +CD8 +细胞在CIK组扩增了近500倍，C＋P组和PersonGen TM 组扩增达千倍；CD3 +CD56 + NKT细胞在CIK组扩增700多倍，其他两组达千倍。杀伤实验结果显示，当E∶T为5∶1时，3组细胞对K562杀伤率为（45.53±19.16）%、（36.53±10.6）%和（53.17±5.83）%，对RPMI 8226杀伤率为（41.23±12.5）%、（38.83±4.3）%和（45.73±7.48）%。结论： Person Gen TM 扩增技术获得的T细胞数量多、纯度高，其中CD3 +CD8 +和CD3 +CD56 + NKT细胞比例较高，其对K562和RPMI 8226肿瘤细胞的杀伤效果相似于CIK细胞扩增方案获得的T细胞。

Objective: This study aimed to evaluate the difference between the first/second stimulation signal-based T lymphocyte amplification technology (PensonGenTM) and cytokine-induced killer (CIK) technology in the expansion and differentiation of T cells in vitro. Methods: Peripheral blood mononuclear cells (PBMCs), isolated from 8 healthy volunteers, were expanded using the CIK technology (CIK group), PersonGen TM human T lymphocyte activation technology (PersonGen TM group), and combined techniques of PensonGen TM and CIK method (C+P group), respectively. The proliferative activity and T cell subsets in different groups of cells were assessed by hemocytometer counting and flow cytometry analysis. The cytotoxic activities of the expanded T cells were examined in vitro using myeloma RPMI 8226 and leukemia K562 cells as targets. Results: After three weeks of culture, cells in all three groups (CIK, PersonGen TM , and C+P group) were successfully expanded, the multiples were 254±56, 863±218, and 793.3±395, respectively, and the proportion of CD3 + cells were （51.61±14.95）%, （99.21±0.79）%, and （96.14±5.16）%, respectively. At the end of 3-week culture, CD3 +CD4 + cells were expanded approximately five times in CIK group and160 times in both C+P and PersonGen TM group groups, CD3 +CD8 + cells were expanded nearly 500 times in CIK group and up to 1 000 times in both PersonGen TM and C+P group groups, and CD3 +CD56 + NKT cells were expanded approximately 700 times and close to 1 000 times in the other two groups. At an effector-to-target cell ratio of 5〖DK〗∶1, the cytotoxic activity of the cells expanded by CIK technology, PersonGen TM technology and CIK plus PersonGen TM technologies were （45.53±19.16）%, （36.53±10.6）% and （53.17±5.8)%, respectively, when K562 cells were used as a target, and were （41.23±12.5）%, （38.83±4.3）% and （45.73±7.48）%, respectively, when RPMI 8226 cells were used as a target. The expanded T cells in PersonGen TM group had slight higher cytotoxicity against both K562 and RPMI 8226 cells than cells expanded in CIK group (P>0.05). Conclusion: Compared to CIK technology, secondary stimulation signal-based PersonGen TM technology is more efficient in the expansion of CD3 +CD8 + cell population from PBMCs, resulting in an enhanced anti-tumor activity.

国家自然科学基金资助项目（No. 31471283）；江苏省科技支撑社会发展项目（No. BE2011682）；江苏省高等学校大学生实践创新训练计划项目（No. 201310285101X）。