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  • 杜平,陈秋莉,段芳龄,曹广文,杨淑英.TNF,IFN,MHCⅠ类分子基因“正反馈”式放大表达的重组腺病毒载体的构建[J].第二军医大学学报,1999,20(9):    [点击复制]
  • 杜平,陈秋莉,段芳龄,曹广文,杨淑英.Construction of adenoviral vector increasing tumor immunogenicity by enhancing gene expression of TNF,IFN,MHC Ⅰmolecule[J].Acad J Sec Mil Med Univ,1999,20(9):   [点击复制]
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TNF,IFN,MHCⅠ类分子基因“正反馈”式放大表达的重组腺病毒载体的构建
杜平,陈秋莉,段芳龄,曹广文,杨淑英
0
()
摘要:
目的:为使外源性TNF,IFN基因和内源性主要组织相容性复合物Ⅰ类(MHCⅠ)分子基因表达呈现“正反馈”式放大效应,构建在 HLA-B7启动子调控、IRES连接下协同表达TNFα和IFNβ基因的重组腺病毒载体。方法和结果:从pGL3B7TNF及pIRIF中切下B7pro-TNF和IRES-IFNβ基因片段,先后插入pBluescript Sk (+), 构建成pBLB7TNIRIF。回收TNFα-IRES-IFNβ基因片段,连入AdCMVLink1中,构建成CMV启动子驱动表达的AdCMVTNIRIF(+)。回收B7pro-TNF-IRES-IFNβ基因片段,连入缺失E1和E3区及启动子的AdBglⅡ中, 构建成HLA-B7启动子驱动表达的AdB7TNIRIF(-)。结论:此两种载体为“正反馈”式放大肿瘤免疫原性提供一条新途径。
关键词:  干扰素、肿瘤坏死因子、HLA-B7抗原启动子、腺病毒载体
DOI:
基金项目:
Construction of adenoviral vector increasing tumor immunogenicity by enhancing gene expression of TNF,IFN,MHC Ⅰmolecule
杜平,陈秋莉,段芳龄,曹广文,杨淑英
()
Abstract:
Objective::To increase tumor immunogenicity by enhancing gene expression of TNF,IFN,MHC Ⅰ molecule , adenoviral vectors containing HuTNFα and IFNβ cDNA driven by HLA-B7 inducible promoter and CMV promoter were constructed, respectively. Methods and Results:The 2 fragments HLA-B7 promoter-TNF and IRES-IFN were isolated from and pGL3B7TNF and pIRIF and insert to pBluescript Ⅱ Sk (+) to generate pBLB7TNIRIF.TNFα-IRES-IFNβ was isolated and ligated to AdCMVLink1 to generate AdCMVTNIRIF(+), in which gene transcription was driven by CMV promoter.B7pro-TNFα-IRES-IFNβ was isolated and ligated to AdBglⅡto generate AdB7TNIRIF(-),in which gene expression was driven by HLA-B7 promoter. Conclusion:The constructed vectors can be a new route for enlarging tumor immunogenicity.
Key words:  interferon  tumor necrosis factor  HLA-B7 promoter  adenoviral vector