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