【摘要】 目的:研究人SCF和人IL-15基因共转染NK细胞系的生物学特性。方法:利用LipofectAMINETM将IL-15真核表达载体pcDNA3-hIL15和SCF真核表达载体pcDNA3hSCF共转染NK92细胞,RTPCR鉴定表达SCF和IL-15的细胞克隆并以SCF依赖细胞株TF1和IL-15依赖细胞株CTLL2测定上清活性,MTT法检测增殖和杀伤活性。流式细胞术检测细胞表面分子CD3、CD16、CD56、CD25、CD48、CD54、CD69和CD95。结果:建立共表达hSCF和hIL15基因的NK92S15细胞株,在IL2培养条件下,其增殖能力表现出更强的聚集趋势,且生长要求有一定的细胞浓度,杀伤活性有不同程度下降。细胞表面分子CD16和CD56没有显著变化,而NK细胞活化相关分子CD25、CD48、CD54和CD95明显降低。结论:可以通过SCF和IL15基因共修饰NK92,改变其生物学特性,观察NK细胞活化、杀伤相关分子的特征,使NK细胞系有更深入的研究价值。更多还原

【Abstract】 Objective:To study the characterization of NK cell line co-transfected with hSCF and hIL-15 gene.Methods:pcDNA3-hSCF and pcDNA3-hIL-15 were transfected into NK-92 cell line with LipofectAMINETM,RT-PCR was used to identify NK-92 cell which express hSCF and IL-15,the activity of supernantes was respectively assayed by TF-1 and CTLL-2 cell line. CD3, CD16, CD56, CD25, CD48, CD54, CD69 and CD95 molecules were tested by FACS.Results:We established NK-92S15 cell line which express hSCF and hIL-15 steadily, proliferation ability demonstrated it had more extensive assembling trend,growing with enough cell number,the cytotoxicity of NK-92S15 cell was decreased compared with parental cell line when incubated with rhIL-2.CD56 and CD16 showed on difference while CD25,CD48, CD54 and CD95 decreased significantly.Conclusion:The characterization of NK-92 could be changed by co-transfecting with hSCF and hIL-15 gene, which was benefit to observe the character of NK cell activation and cytotoxicity related molecules. The gene transfection of NK-92 cell made it suitable for further study.更多还原