重组人纤维连接片段联合抗CD3单抗包被对急性白血病CIK增殖和杀伤活性的影响

Effects of Recombinant Human Fibronectin Fragment Combined with Anti-CD3 Monoclonal Antibody on the Proliferation and Cytotoxicity of Cytokine-induced Killer Cells from Acute Leukemia

目的探讨重组人纤维连接片段(RetroNectin)与抗CD3单抗(CD3Ab)联合包被培养对细胞因子诱导的杀伤细胞(CIK)增殖和杀伤活性的影响。方法取完全缓解期急性白血病(AL)患者外周血单个核细胞,采用RetroNectin包被(RN组)、CD3Ab包被(CD3Ab组)及RetroNectin联合CD3Ab包被(RN+CD3Ab组)与传统方法培养(对照组)获得CIK细胞,比较各组增殖、表达及杀伤活性。结果 CIK扩增:各实验组CIK扩增倍数高于对照组,且RN+CD3Ab组高于RN组和CD3Ab组(P<0.05)。收获细胞CD25+表达:RN组和RN+CD3Ab组高于对照组和CD3Ab组(P<0.05)。各期细胞比例:RN组和RN+CD3Ab组的G1期细胞比例低于CD3Ab组和对照组,而S期细胞高于CD3Ab组和对照组(P<0.05)。CIK对自体AL细胞的杀伤活性:效靶比10∶1、20∶1和40∶1时,RN组和RN+CD3Ab组高于对照组和CD3Ab组(P<0.05)。收获细胞凋亡率:RN组和RN+CD3Ab组较CD3Ab组和对照组下降(P<0.05)。结论 RetroNectin联合CD3Ab包被用于CIK体外培养可获得增殖率和活性更高的免疫活性细胞,是一种值得推荐的方法。

Objective To investigate the effects of recombinant human fibronectin fragment （RetroNectin） combined with anti-CD3 monoclonal antibody （CD3Ab） on the proliferation and cytotoxicity of cytokine-induced killer cells （CIK） from acute leukemia （AL）. Methods Mononuclear ceils （MNCs） were isolated from peripheral blood of complete remission AL pa- tients. The MNCs were cultured in vitro by precoating with RetroNecn （RN group）, CD3Ab （CD3Ab group）, RetroNectin com- bined with CD3Ab （RN＋CD3Ab group） and traditional method （control group） to generate CIK. The changes of growth rate, characterization, cytotoxicity and apoptosis of CIK were determined between groups. Results The amplification of CIK was higher in experimental group than that of control group, and the amplification of CIK was higher in group RN＋CD3Ab than that of in group RN and group CD3Ab （P 〈 0.05）. The expression of CD25 positive cells was higher in group RN and group RN＋CD3Ab than that of group CD3Ab and control group （P 〈 0.05）.The percentage of GI stage cells was lower in group RN and group RN＋CD3Ab than that of group CD3Ab and control group. The percentage of S stage cells was higher in group RN and group RN＋CD3Ab than that of group CD3Ab and control group （P 〈 0.05）. The cytotoxicity was higher in group RN and group RN＋CD3Ab than that of group CD3Ab and control group （P 〈 0.05） at the E/T scope 40 ： 1.The percentage of apoptotic cells was lower in group RN and group RN＋CD3Ab than that of group CD3Ab and control group （P 〈 0.05）. Conclusion These in vitro studies suggest that a higher activity of immune cells could be obtained by CIK cells cultured by precoating Ret- roNectin and CD3Ab.