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Technigue and Method
PersonGen TM versus CIK induction techniques in the expansion and activation of human T cells
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Abstract:
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.
Keywords:
Project Supported:
Project supported by the National Natural Science Foundation of China（No. 31471283）, the Science and Technology Support Program of Jiangsu Province（No. BE2011682）, and the Jiangsu Student Innovation and Entrepreneurship Training Program（No. 201310285101X）
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