hKDR基因胞外区mRNA转染树突状细胞诱发对表达mKDR的Hepa1-6细胞的特异性杀伤效应  

Immunity elicited by dendritic cells modified by mRNA encoding a truncated hKDR gene product against tumor cells expressing mKDR in vitro

作　　者：孙林英[1] 刘畅[2] 刘秀丽[3] 邵晓婷[3] 谭晓华[3] 

机构地区：[1]泰山医学院医学检验系 [2]北京军区总医院肿瘤诊疗中心 [3]北京军区总医院血液科

出　　处：《免疫学杂志》2009年第5期495-499,共5页Immunological Journal

基　　金：国家自然科学基金资助项目(30371627,30271185)

摘　　要：目的探讨人血管内皮细胞生长因子受体-2(KDR)胞外段(Ig1~3)mRNA修饰的树突状细胞(DC)诱发的体外特异性细胞免疫效应。方法构建pmRNA IRES-hKDR)(Ig1~3),体外转录出相应的mRNA,Western blot鉴定其蛋白表达;以hKDR(Ig1~3)mRNA致敏来源于小鼠骨髓的DC,并免疫小鼠,同时设空白DC对照组,7d后取鼠脾细胞为效应细胞,表达mKDR的肿瘤细胞为靶细胞按100∶1、50∶1、25∶1的比例混合,用乳酸脱氢酶(LDH)释放法检测细胞毒作用。结果未经修饰DC激活的细胞毒性T淋巴细胞(CTL)对靶细胞的杀伤率为(19.6±3.7)%、(10.2±2.6)%、(6.2±2.2)%,经mRNA修饰的DC激活的CTL对靶细胞杀伤率为(75.2±2.3)%、(54.3±3.4)%、(20.9±3.1)%,同一效靶比情况下显著强于其他3组。结论hKDR(Ig1~3)mRNA致敏的DC能诱导针对Hepa1-6(mKDR)的特异性细胞免疫反应。Objective To investigate the specific cytotoxic T lymphocytes in vitro,which activated by dendritic cells loaded with mRNA encoding human extracellular domain of vascular endothelial growth factor receptor-2.Methods The vector pmRNA IRES-hKDR(Ig1-3)was constructed via molecular cloning methods.Protein expression of hKDR(Ig1-3)was detected in transfected COS-7 cells by Western blotting.Mice were immunized with syngeneic DCs transfected with hKDR(Ig1-3)mRNA.Splenocytes were harvested 7 days after the immuniza...

关 键 词：血管内皮细胞生长因子受体2 树突状细胞 细胞毒性T淋巴细胞反应 

分 类 号：R392.1[医药卫生—免疫学]