不同磷酸化合物扩增外周血γδ T细胞的效率及条件优化  

Amplification efficency and optimization of culture conditions of γδ T cells in peripheral blood by different phosphate compounds

作　　者：习燕[1] 苗天雨 万浬科[1] 王元虎[1] 冯婷 巩天祥[1] 李明远[3] 

机构地区：[1]成都市血液中心 [2]四川大学华西临床医学院 [3]四川大学华西基础医学与法医学院,四川成都610041

出　　处：《细胞与分子免疫学杂志》2014年第8期868-871,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81170606);四川大学大学生创新创业计划(201310610068)

摘　　要：目的研究异戊烯焦磷酸(IPP)与帕米磷酸钠(PAM)体外扩增成人外周血γδT细胞的效率及条件优化。方法梯度离心法分离人外周血单个核细胞(PBMC),分别加入(1.0、5.0、10.0、15.0)μg/mL IPP或(2.0、5.0、8.0、12.0)μg/mL PAM及(100.0、200.0、500.0)IU/mL IL-2,置于100 mL/L小牛血清的RPMI1640培养体系培养,观察并收集细胞,采用流式细胞仪检测刺激后CD3+TCRδ2+的γδT细胞数量以及比例,并计算其扩增效率。结果经过14 d扩增培养后,IPP组的γδT细胞数量在淋巴细胞中比例可上升为81.3%,PAM组的γδT细胞数量在淋巴细胞中比例上升为78.5%,表明IPP和PAM均能有效刺激扩增γδT细胞,2组的扩增效率无统计学差异(P>0.05)。结论 PAM具有与IPP相似的体外扩增γδT细胞的能力,PAM扩增法可能成为扩增γδT细胞更为经济实用的选择。Objective To compare the efficiency of pamidronate （PAM） and isopentenyl pyrophosphate （IPP） to stimulate γδT cell expansion from human peripheral blood and explore the optimized expansion conditions. Methods Peripheral blood mononuclear cells （PBMCs） were isolated by FicolI-Paque gradient centrifugation, and then cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, IPP （1.0, 5,0, 10,0, 15,0 μg/mL） or PAM （2.0, 5,0, 8.0, 12.0 μg/mL）, and IL-2 （100.0,200.0, 500.0 IU/mL）. The cells were observed and collected. The number and proportion of CD3 ＋ TCRδ2 ＋ γδT cells stimulated by PAM or IPP in total lymphocytes were evaluated by flow cytometry and the expansion efficiency was calculated. Results After 14 days, the ratios of γδT cells in total lymphocytes in IPP group and PAM group increased to 81.3% and 78.5%, respectively. This indicated that both IPP and PAM could effectively stimulate γδT cell expansion and there was no significant difference in the efficiency of expansion between the two groups （ P 〉 0.05）. Conclusion PAM has the similar ability with IPP to stimulate γδT cell expansion in vitro. PAM could become more economical and practical choice for stimulating γδT cell expansion.

关 键 词：ΓΔT细胞 异戊烯焦磷酸 帕米磷酸钠 体外扩增 

分 类 号：R392.11[医药卫生—免疫学] Q253[医药卫生—基础医学]