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4-1BBL重组腺病毒载体的构建及表达

Construction and expression of 4-1BBL recombinant adenovirus vector
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摘要 目的构建4-1BBL基因重组腺病毒载体,为前列腺癌的免疫治疗奠定基础。方法以质粒pcDNA3-m4-1BBL为模板,PCR法扩增出4-1BBL cDNA,并将其插入腺病毒穿梭质粒pAdTrack-CMV中构建成腺病毒穿梭质粒pAdTrackCMV-m4-1BBL,经PmeⅠ酶切线性化,与腺病毒骨架质粒pAdEasy-1共转化大肠杆菌BJ5183,挑选同源重组质粒,PacⅠ酶切线性化后,转染HEK293细胞包装成重组病毒颗粒,荧光显微镜观察绿色荧光表达,RT-PCR和Western blot法检测4-1BBL表达。结果目的基因经酶切分析,测序鉴定正确,RT-PCR和Western blot证实4-1BBL表达。结论成功构建了含4-1BBL基因的腺病毒载体,该载体可用于前列腺癌的免疫治疗。 Objective To construct recombinant adenovirus vector encoding 4-1BBL gene and pave the way for the immunotherapy of prostate cancer.Methods Amplified the 4-1BBL cDNA fragment from plasmid pcDNA3-m4-1BBL by PCR method,and inserted it into adenovirus shuttle plasmid pAdTrack-CMV resulting pAdTrackCMV-m4-1BBL.Then Pme Ⅰ-linearized plasmid pAdTrackCMV-m4-1BBLand co-transformed into competent BJ5183 with adenovirus backbone plasmid pAdEasy-1.After homologous recombination in BJ5183,pAd-m4-1BBL was linearized by Pac Ⅰ,and transfected into HEK293 cells.Then Ad-m4-1BBL was packaged and amplified.GFP was observed by fluorescenct microscope,and 4-1BBL expression was detected by RT-PCR and Western blot.Results Ad-m4-1BBL was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis.4-1BBL expression was confirmed by RT-PCR and Western blot.Conclusion Recombinant adenovirus vector carrying 4-1BBL is successfully constructed and can be used for the immunotherapy of prostate cancer.
作者 陈国晓 匡幼林 翁小东 陈志远 祝恒成 江波涛 刘修恒
机构地区 武汉大学人民医院
出处 《山东医药》 CAS 北大核心 2010年第19期16-18,共3页 Shandong Medical Journal
基金 国家自然科学基金资助项目(30672107)
关键词 4-1BBL 重组腺病毒 基因治疗 4-1BBL recombinant adenovirus gene therapy
分类号 R392.33 [医药卫生—免疫学]
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参考文献9

  • 1Cheuk AT,Mufti GJ,Guinn BA.Role of 4-1BB:4-1BB ligand in cancer immunotherapy[J].Cancer Gene Ther,2004,11(3):215-226.
  • 2Kwon B,Lee HW,Kwon BS.New insights into the role of 4-1BB in immune responses:beyond CD8^+ T cells[J].Trends Immunol,2002,23(8):378-380.
  • 3Watts TH.Tnf/Tnfr family members in costimulation of T cell responses[J].Annu Rev Immunol,2005,23(1):23-68.
  • 4Dawicki W,Watts TH.Expression and function of 4-1BB during CD4 versus CD8 T cell response in vivo[J].Eur J Immunol,2004,34(3):743-751.
  • 5Nam KO,Kang H,Shin SM,et al.Cross-linking of 4-1BB activates TCR-signaling pathways in CD8^+ T lymphocytes[J].J Immunol,2005,174(4):1898-1905.
  • 6Chie KS,James WH,Heesun K,et al.4-1BB ligand enhances tumor-specific immunity of poxvirus vaccines[J].Vaccine,2006,24(6):4975-4986.
  • 7Guo HX,Jiang WG,Liu WG,et al.Extracellular domain of 4-1BBL enhanced the antitumoral efcacy of peripheral blood lymphocytes mediated by anti-CD3 xanti-Pgp bispecifc diabody against human multidrug-resistant leukemia[J].Cellular Immunology,2008,251(3):102-108.
  • 8Fisher KI,Choi H,Burda I,et al.Recombinant adenovirus deleted of all viral genes for gene therapy of cystic fibrosis[J].Virology,1996,217(1):11-22.
  • 9张俊,王爱东,申咏梅,石怡珍,崔学军,刘增礼,欧阳松应.hTERT核心启动子调控的hNIS重组腺病毒的构建与鉴定[J].中国免疫学杂志,2008,24(6):549-553. 被引量：3

二级参考文献13

  • 1Dingli D, Diaz R M, Bergert E R et al. Genetically targeted radiotherapy for multiple myeloma[J]. Blood,2003 ; 102(2) :489-496.
  • 2Scholz I V, Cengic N, Baker C H et al. Radioiodine therapy of colon cancer following tissue-specific sodium iodide symporter gene transfer [ J ]. Gene Ther, 2005 ; 12 (3) : 272-280.
  • 3Cengic N, Baker C H, Schutz M et al. A novel therapeutic strategy for medullary thyroid cancer based on radioiodine therapy following tissue- specific sodium iodide symporter gene expression [ J]. J Clin Endocrinol Metab, 2005 ; 90(8) :4457-4464.
  • 4Feng J, Funk W D, Wang S S et al. The RNA component of human telomerase[J]. Science, 1995;269(5228) : 1236-1241.
  • 5Gu J, Kagawa S, Takakura M et al. Tumor-specific transgene expression from the human telomerase reverse transcriptase promoter enables targeting of the therapeutic effects of the Bax gene to cancers[J]. Cancer Res, 2000; 60 ( 19 ) : 5359-5364.
  • 6Majumdar A S, Hughes D E, Lichtsteiner S P et al. The telomerase reverse transcriptase promoter drives efficacious tumor suicide gene therapy while preventing hepatotoxicity encountered with constitutive promoters [ J ]. Gene Ther,2001 ;8(7) :568-578.
  • 7Dai J,Rabie A B,Hagg U et al. Alternative gene therapy strategies for the repair of craniofacial bone defects[ J ]. Curr Gene Ther, 2004 ; 4 (4) : 469-485.
  • 8Mitani K, Kubo S. Adenovirus as an integrating vector[J]. Curr Gene Ther, 2002, 2(2) : 135-144.
  • 9Cao B, Mytinger J R, Huard J. Adenovirus mediated gene transfer to skeletal muscle[ J]. Microscopy Research and Technique, 2002; 58 ( 1 ) : 45-51.
  • 10He T C,Zhou S,da Costa L T et al. A simplified system for generating recombinant adenoviruses[ J]. Proc Natl Acad Sci USA, 1998; 95 (5) : 2509-2514.

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山东医药
2010年 第19期

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