人外周血细胞毒性CD3^-CD56^+ NK细胞高效扩增的研究  

Study on efficient expansion of human cytotoxic CD3^-CD56^+ NK cells from peripheral blood

作　　者：黄朝晖[1] 王丰[1] 刘志辉[1] 华东[1] 李莉华[1] 任金冬[1] 

机构地区：[1]苏州大学附属第四医院肿瘤研究所,江苏无锡214062

出　　处：《肿瘤防治研究》2004年第1期36-38,共3页Cancer Research on Prevention and Treatment

基　　金：无锡市卫生局"十五"重大科研项目资助课题

摘　　要：目的　探索从人PBMC中高效扩增细胞毒性CD3-CD5 6 + NK细胞的方法。方法　使用干细胞生长培养基 (SCGM )和RPMI 16 4 0培养基 ,在抗CD3单抗、IL 2和植物血凝素 (PHA)的作用下从 5例健康成人PBMC中诱导扩增CD3-CD5 6 + NK细胞 ,并用MTT法检测其细胞毒活性。结果　只有在使用SCGM为基础培养基时 ,PBMC经抗CD3单抗、IL 2作用获得大量增殖 ,在PHA存在时获得最大增殖(P <0 .0 5 ) ,在 14天时扩增 5 1.3± 7.2倍 ,含有 (5 2 .4± 7.9) %CD3-CD5 6 + NK细胞和 (14 .2± 4 .0 ) %CD3+ CD5 6 + T细胞 ;在效 /靶比 10∶1时 ,对K5 6 2和Raji的杀伤率分别达83.7%和 5 5 .8%。结论　可使用SCGM ,在抗CD3单抗、IL 2和PHA协同作用下大量扩增细胞毒性CD3-CD5 6 + NK细胞 ,为应用NK细胞进行肿瘤过继免疫治疗提供了一种简单有效的扩增NK细胞的方法。Objective To study the expanding way of human cytotoxic CD3 -CD56 + NK cells from peripheral blood mononuclear cells PBMC. Methods PBMCs were cultured in stem cell growth medium SCGM or RPMI 1640 supplemented with monoclonal anti-CD3 antibodies, IL-2 and PHA at varying concentrations. The cytotoxicity of the expanded cells was detected by MTT method. Results After being activated by monoclonal anti-CD3 antibodies and IL-2, PBMCs cultured in SCGM get the obvious expansion, performed the highest proliferation while PHA was present( P < 0.05 ), expanded 51.3 ± 7.2 fold and contained ( 52.4 ± 7.9 )% CD3 -CD56 + NK cells, ( 14.2 ± 4.0 )% CD3 +CD56 +T cells. The expanded cells population lysed 83.7% of K562 and 55.8% of Raji targets in a 10∶1 effector to target ratio, respectively. Conclusion Cytotoxic cells including major NK cells could be expanded from PBMC using SCGM with the synergistic inducing by monoclonal anti-CD3 antibodies, IL-2 and PHA. This work provided a simple and efficient way of expanding and enriching human NK cells for the adoptive immunotheraphy of tumor with NK cells.

关 键 词：人外周血 细胞毒性 CD3^-细胞 CD56^+细胞 NK细胞 检测 细胞活性 肿瘤 免疫治疗 

分 类 号：R730.51[医药卫生—肿瘤]