title: [Transfection Efficiency of Ad5F11p-GFP on CIK and NK-92 Cells and Its Influence on Biological Characteristics].
abstract: OBJECTIVE
  To study transfection efficiency of Ad5F11p-GFP and its influence on biological characteristics of CIK and NK-92 cells in order to predict the application of Ad5F11p vector in immunotherapy.
  
  
  METHODS
  Two kinds of immune cells, cytokine-induced killer (CIK) cells and natural-killer (NK) cell line NK-92 cells, were transfected by Ad5F11p-GFP at different multiplicity of transfection (MOI), and untransfected immune cells were used as negative control. GFP expression was determined by flow cytometry, the cell morphology was observed with microscope, the cell proliferation was analyzed by trypan blue staining, specific cytotoxicity of NK-92 cells was determined by LDH assay.
  
  
  RESULTS
  About 90% of transfection efficiency for NK-92 cells could be achieved at a MOI of 25, while the transfection efficiency for CIK was less than 40% at a MOI of 200. In addition, the transfection efficiency basically unchanged at the same MOI for 48 h and 96 h, and the immune cells transfected with the virus trended to form agglomeration, displaying slower proliferation, increase of IFN-γ release and enhancement of tumor killing activity.
  
  
  CONCLUSIONS
  Ad5F11p- modified NK-92 shows a good prospect for adoptive immunotherapy.
authors.affiliation: Department of Pharmaceutical Engireering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072.
authors.forename: Zan-Mei
authors.initials: ZM
authors.lastname: Xu
authors.affiliation: No. 307 Hospital of Chinese PLA, Beijing 100039.
authors.forename: Ying
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Institute of Biotechnology Medicine, Academy of Military Medical Sciences, Beijing 100850.
authors.forename: Lan-Jun
authors.initials: LJ
authors.lastname: Zhao
authors.affiliation: Institute of Biotechnology Medicine, Academy of Military Medical Sciences, Beijing 100850.
authors.forename: Jin
authors.initials: J
authors.lastname: Liu
authors.affiliation: Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071.
authors.forename: Xian-Wen
authors.initials: XW
authors.lastname: Hu
authors.affiliation: Institute of Biotechnology Medicine, Academy of Military Medical Sciences, Beijing 100850.
authors.forename: Chu-Tse
authors.initials: CT
authors.lastname: Wu
authors.affiliation: Institute of Biotechnology Medicine, Academy of Military Medical Sciences, Beijing 100850. E-mail: duanhf0720@163.com.
authors.forename: Hai-Feng
authors.initials: HF
authors.lastname: Duan
mesh_terms: D000256:Adenoviridae; D002460:Cell Line; D049109:Cell Proliferation; D055612:Cytokine-Induced Killer Cells / Q000166:cytology*; D003602:Cytotoxicity, Immunologic; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016219:Immunotherapy, Adoptive; D007694:Killer Cells, Natural / Q000166:cytology*; D009369:Neoplasms / Q000628:therapy; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins