天津医药

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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

李青1,邓琦2,刘鹏江1,白雪1,李玉明1   

  1. 1. 天津市第一中心医院血液科
    2. 天津市第一中心医院 血液科
  • 收稿日期:2012-06-27 修回日期:2013-02-22 出版日期:2013-07-15 发布日期:2013-07-15
  • 通讯作者: 邓琦

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

liqing ,DENG Qi ,LIU Pengjiang ,BAI Xue ,LI Yuming   

  1. Department of Hematology, the First Central Hospital of Tianjin, Tianjin 300192, China
  • Received:2012-06-27 Revised:2013-02-22 Published:2013-07-15 Online:2013-07-15
  • Contact: DENG Qi

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摘要：

【摘要】 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。

关键词: 重组融合蛋白质类, 纤连蛋白类, 抗原, CD3, 抗体, 单克隆, 细胞因子类, 杀伤细胞

Abstract: Objective   To investigate the effects of precoated with retroNectin and CD3Ab on the proliferation and cytotoxicity of cytokine-induced killer cells (CIK) from acute leukemia. Methods   Mononuclear cells (MNCs) were isolated from peripheral blood of complete remission AL patients. The MNCs were cultured in vitro precoated with retroNectin(groupⅠ), CD3Ab(groupⅡ), retroNectin combined with CD3Ab(groupⅢ) and cultured by traditional method(control group) to generate CIK. The change of growth rate, phenotypic characterization, secretion of cytokines of CIK, cell cycle, cytotoxicity and apoptosis of CIK were determined. Result     ① The amplification of CIK in groupⅠ（173.19 times）,Ⅱ（153.79 times）and Ⅲ（224.75 times） were higher than that of in control group（110.23 times）, and the amplification of CIK in group Ⅲ was higher than that of in groupⅠ,Ⅱ（P<0.05）. ② The expression of CD25 positive cells in groupⅠ（39.84±8.25）% and Ⅲ（42.16±10.38）% were higher than that of in groupⅡ（11.93±3.47）% and control group（13.76±2.52）%（P<0.01）. ③The percentage of G1 stage cells in groupⅠ（56.92±9.55）% and Ⅲ（54.84±8.83）% were lower than that of in groupⅡ（69.86±7.72）% and control group（69.72±8.44）%, and the percentage of S stage cells were higher（P<0.05）. ④The cytotoxicity in groupⅠ(63.44±12.75)% and Ⅲ（65.02±12.34）% were higher than that of in groupⅡ(50.13±10.53)% and control group(47.96±7.73)%（P<0.05）at the E/T scope 40:1.⑤The percentage of apoptosis cells in groupⅠ（8.9%）and Ⅲ（9.0%） were lower than that of in groupⅡ（35.1%） and control group（25.4%）（P<0.05）. Conclusion   These in vitro studies suggest that we could obtain CIK cells that the proliferation and cytotoxicity were more higher by precoated with retroNectin and CD3Ab.

Key words: recombinant fusion proteins, fibronectins, antigens, CD3, antibodies, monoclonal, cytokines, 杀伤细胞