摘要
目的探讨不同形式IL15基因转染对人小细胞肺癌细胞株NCIH446细胞HLA的Ⅰ类和Ⅱ类分子表达及NK杀伤敏感性的影响。方法在本室已构建的3种细胞模型(转染IL15成熟肽基因的NCIH446/IL15mp细胞、转染原型IL15基因的NCIH446/IL15细胞和转染以IL2信号肽替代IL15信号肽的改型IL15基因的NCIH446/IL2spIL15mp细胞)基础上[1],以野生株细胞为对照,流式细胞分析仪(Fluorescenceactivatedcellsorter,FACS)分析这3种细胞HLA的Ⅰ类和Ⅱ类分子表达,四甲基偶氮唑蓝(MTT)法测定它们对NK杀伤的敏感性。结果野生株NCIH446细胞表面阳性表达HLA的Ⅰ类和Ⅱ类分子;转染IL15成熟肽基因或改型IL15基因后,这两种分子的表达均被抑制为阴性;转染原型IL15基因后,这两种分子的表达均被显著上调。5名不同健康志愿者的外周血单个核细胞(PBMC)对野生株NCIH446细胞的平均NK杀伤率最低,对NCIH446/IL15、NCIH446/IL15mp和NCIH446/IL2spIL15mp细胞的平均NK杀伤率依次升高。结论转染不同形式IL15基因对NCIH446细胞的免疫生物学特性影响不同。转染IL15成熟肽基因或改型IL15基因,抑制NCIH446细胞表面HLA的Ⅰ类和Ⅱ类分子表达;转染原型IL15基因,上调这两种分子的表达;转染不同形式IL15基因均可增强NCIH446细胞对NK杀伤的敏感性,但程度不同:以改型IL15者最高,IL15成熟肽者次之,原型IL15者最低。
Objective To investigate the effects of transfecfion of IL-15 gene into NCI-H446 on the expression of HLA-Ⅰ and HLA-Ⅱ and the sensitivity to NK cytotoxicity. Methods The HLA-I and HLA-Ⅱ expressions of all these transfected cells were analyzed by FACS and their sensitivities to NK cytotoxicity were determined by MTT, respectively. Results The expressions of HLA-Ⅰ and HLA-Ⅱ on NCI-H446/IL-15 were higher than those on NCI-H446. FACS indicated that the transfection of IL-15mp or IL-2sp-IL-15mp could inhibit the expressions of HLA-Ⅰ and HLA-Ⅱ. In 5 health volunteers, the killing rate to NCI-H446 was the lowest, and the killing rates NCI-H446/IL-15, NCI-H446/IL-15mp, and NCI-H446/IL-2sp-IL-15mp were gradually increased. Conclusion The effects of different IL-15 gene transfection on the immunobiological characteristics of NCI-H446 are different. Either IL-15mp or IL-2sp-IL-15mp gene is able to inhibit the expressions of HLA-Ⅰ and HLA-Ⅱ, but the prototypic IL-15 gene could up-regulate the expression. The sensitivity of NCI-H446/IL-2sp-IL-15mp to NK cytotoxicity is the highest, and that of NCI-H446/IL-15 is lower than NCI-H446/IL-15mp but higher than NCI-H446.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2005年第6期468-470,共3页
Immunological Journal
基金
河北省自然科学基金资助项目(302499)
